AU2008212020A1 - Nucleic acids and corresponding proteins entitled 109P1D4 useful in treatment and detection of cancer - Google Patents

Description

I

AUSTRALIA

Patents Act 1990 FB RICE CO Patent and Trade Mark Attorneys AGENSYS, INC.

COMPLETE SPECIFICATION STANDARD PATENT Invention Title: Nucleic acids and corresponding proteins entitled 109P1D4 useful in treatment and detection of cancer The following statement is a full description of this invention including the best method of performing it known to us:- 00 1A NUCLEIC ACIDS AND CORRESPONDING PROTEINS ENTITLED 109P1 04 USEFUL IN TREATMENT AND DETECTION OF CANCER RELATED APPLICATIONS V)This application is a divisional application under S.79B of the Patents Act 1990 of Australian Patent Application No.

S 5 2004235755 filed 30 April 2004 which corresponds to International Application No. PCT/US2004/013568 in the Australian national phase, and claims priority from United States Patent Application No. 60/467,002 filed on 30 April 2003. The contents of each of the foregoing applications is hereby incorporated in its entirety by way of reference into this divisional application.

FIELD OF THE INVENTION 00 The invention described herein relates to genes and their encoded proteins, termed 109P1D4 and variants thereof, expressed in certain cancers, and to diagnostic and therapeutic methods and compositions useful in the management of cancers that express 109P1D4.

BACKGROUND OF THE INVENTION Cancer is the second leading cause of human death next to coronary disease. Worldwide, millions of people die from cancer every year. In the United States alone, as reported by the American Cancer Society, cancer causes the death of well over a half-million people annually, with over 1. 2 million new cases diagnosed per year. While deaths from heart disease have been declining significantly, those resulting from cancer generally are on the rise. In the early part of the next century, cancer is predicted to become the leading cause of death.

Worldwide, several cancers stand out as the leading killers. In particular, carcinomas of the lung, prostate, breast, colon, pancreas, and ovary represent the primary causes of cancer death. These and virtually all other carcinomas share a common lethal feature. With very few exceptions, metastatic disease from a carcinoma is fatal. Moreover, even for those cancer patients who initially survive their primary cancers, common experience has shown that their lives are dramatically altered. Many cancer patients experience strong anxieties driven by the awareness of the potential for recurrence or treatment failure. Many cancer patients experience physical debilitations following treatment. Furthermore, many cancer patients experience a recurrence.

Worldwide, prostate cancer is the fourth most prevalent cancer in men. In North America and Northemrn Europe, it is by far the most common cancer in males and is the second leading cause of cancer death in men. In the United States alone, well over 30, 000 men die annually of this disease-second only to lung cancer. Despite the magnitude of these figures, there is still no effective treatment for metastatic prostate cancer. Surgical prostatectomy, radiation therapy, hormone ablation therapy, surgical castration and chemotherapy continue to be the main treatment modalities.

Unfortunately, these treatments are ineffective for many and are often associated with undesirable consequences.

On the diagnostic front, the lack of a prostate tumor marker that can accurately detect early-stage, localized tumors remains a significant limitation in the diagnosis and management of this disease. Although the serum prostate specific antigen (PSA) assay has been a very useful tool, however its specificity and general utility is widely regarded as lacking in several important respects.

00 Progress in identifying additional specific markers for prostate cancer has been improved by the generation of prostate cancer xenografts that can recapitulate different stages of the disease in mice. The LAPG (Los Angeles Prostate CK1 Cancer) xenografts are prostate cancer xenografts that have survived passage in severe combined immune deficient (SCiD) mice and have exhibited the capacity to mimnic the transition from androgen dependence to androgen Independence (Klein et A, 1997, Nat. Med. 3:402). More recently identified prostate cancer markers indlude PCTA-1 (Su et al., 1996, Proc. Nall.

Acad. Sci. USA 93: 7252), prostate-specific membrane (PSM) antigen (Pinto et Olin Cancer Res 1996 Sep 2 1445- 51), STEAP (Hubert, et Proc Nail Acad Sci U S A 1999 Dec 7; 96(25): 14523-8) and prostate stem cell antigen (PSCA) (Reiter et 1998, Proc. Nall. Acad. Sci. USA 95: 1735).

While previously identified markers such as PSA, PSM, PCTA and PSCA have facilitated efforts to diagnose and treat prostate cancer, there Is need for the identification of additional markers and therapeutic targets for prostate and related cancers in order to further improve diagnosis and therapy.

CK1 Renal cell carcinoma (ROC) accounts for approximately 3 percent of adult malignancies. Once adenomas reach a diameter 0 and transitional cell carcinoma of the renal pelvis or ureter. The Incidence of renal cell adenocarcinoma Is estimated at more than 29,000 cases in the United States, and more than 11,600 patients died of this disease in 1998. Transitional cell carcinoma Is less frequent with an Incidence of approximately 500 cases per year In the United States.* Surgery has been the primary therapy for renal cell adenocarcinona for many decades. Until recently, metastatic disease has been refractory to any systemic therapy. With recent developments in systemic therapies, particularly immunotherapies, metastatic renal cell carcinoma may be approached aggressively in appropriate patients with a possibility of durable responses. Nevertheless, there Is a remaining need for effective therapies for these patients.

Of all new cases of cancer in the United States, bladder cancer represents approximately 5 percent in men (fifth most common neoplasm) and 3 percent in women (eighth most common neoplasm). The Incidence Is increasing slowly, concurrent with an increasing cider population. In 1998, there was an estimated 54,500 cases, including 39,500 In men and 15,000 in women. The age-adjusted incidence in the United States is 32 per 100,000 for men and eight per '100,000 in women. The historic malelfemale ratio of 3:1 may be decreasing related to smoking patterns in women. There were an estimated 11,000 deaths from bladder cancer in 1998 (7,800 in men and 3,900 in women). Bladder cancer Incidence and mortality strongly increase with age and will be an Increasing problem as the population becomes more elderly.

Most bladder cancers recur In the bladder. Bladder cancer Is managed with a combination of transurethral resection of the bladder (TUR) and intravesical chemotherapy or immunotherapy. The multifocal and recurrent nature of bladder cancer points out the limitations of 11JR. Most muscle-Invasive cancers are not cured by TUR alone. Radical cystectomy and urinary diversion Is the most effective means to eliminate the cancer but carry an undeniable Impact on urinary and sexual function. There continues to be a significant need for treatment modalities that are beneficial for bladder cancer patents.

An estimated 130,200 cases of colorectall cancer occurred In 2000 in the United States, Including 93,800 cases of colon cancer and 36,400 of rectal cancer. Colorectall cancers are the third most common cancers in men and women.

Incidence rates declined signlIficantly during 1992-1996 %per year). Research suggests that these declines have been due to Increased screening and polyp removal, preventing progression of polyps to tInvasIve cancers. There were an estimated 56,300 deaths (47,700 from colon cancer, 8,600 from rectal cancer) in 2000, accounting for about 11I% of all U.S.

cancer deaths.

Al present surgery is the most common form of therapy for colorectal cancer, and for cancers that have not spread, It Is frequenly curative. Chemotherapy, or chemotherapy plus radiation, is given before or after surgery to most 00 patients whose cancer has deeply perforated the bowel wall or has spread to the lymph nodes. A permanent colostomy 0 (creation of an abdominal opening for elimination of body wastes) is occasionally needed for colon cancer and is infrequently Crequired for rectal cancer. There continues to be a need for effective diagnostic and treatment modalities for colorectal eC cancer.

There were an estimated 164,100 new cases of lung and bronchial cancer in 2000, accounting for 14% of all U.S.

Scancer diagnoses. The Incidence rate of lung and bronchial cancer is declining significantly in men, from a high of 86.5 per O 100,000 in 1984 to 70.0 in 1996. In the 1990s, the rate of increase among women began to slow. In 1996, the incidence rate in women was 42.3 per 100,000.

SLung and bronchial cancer caused an estimated 156,900 deaths in 2000, accounting for 28% of all cancer deaths.

During 1992-1996, mortality from lung cancer declined significantly among men per year) while rates for women were r still significantly increasing per year). Since 1987, more women have died each year of lung cancer than breast Ccancer, which, for over 40 years, was the major cause of cancer death in women. Decreasing lung cancer incidence and 00 mortality rates most likely resulted from decreased smoking rates over the previous 30 years; however, decreasing smoking Spatterns among women lag behind those of men. Of concern, although the declines in adult tobacco use have slowed, tobacco use in youth is increasing again.

Treatment options for lung and bronchial cancer are determined by the type and stage of the cancer and include surgery, radiation therapy, and chemotherapy. For many localized cancers, surgery is usually the treatment of choice.

Because the disease has usually spread by the time it is discovered, radiation therapy and chemotherapy are often needed in combination with surgery. Chemotherapy alone or combined with radiation is the treatment of choice for small cell lung cancer; on this regimen, a large percentage of patients experience remission, which in some cases is long lasting. There is however, an ongoing need for effective treatment and diagnostic approaches for lung and bronchial cancers.

An estimated 182,800 new invasive cases of breast cancer were expected to occur among women In the United States during 2000. Additionally, about 1,400 neW cases of breast cancer were expected to be diagnosed in men in 2000.

After increasing about 4% per year in the 1980s, breast cancer incidence rates in women have leveled off In the 1990s to about 110.6 cases per 100,000.

In the U.S. alone, there were an estimated 41,200 deaths (40,800 women, 400 men) in 2000 due to breast cancer.

Breast cancer ranks second among cancer deaths in women. According to the most recent data, mortality rates declined significantly during 1992-1996 with the largest decreases In younger women, both white and black. These decreases were probably the result of earlier detection and improved treatment.

Taking into account the medical circumstances and the patient's preferences, treatment of breast cancer may involve lumpectomy (local removal of the tumor) and removal of the lymph nodes under the arm; mastectomy (surgical removal of the breast) and removal of the lymph nodes under the arm; radiation therapy; chemotherapy; or hormone therapy.

Often, two or more methods are used in combination. Numerous studies have shown that, for early stage disease, long-term survival rates after lumpectomy plus radiotherapy are similar to survival rates after modified radical mastectomy. Significant advances in reconstruction techniques provide several options for breast reconstruction after mastectomy. Recently, such reconstruction has been done at the same time as the mastectomy.

Local excision of ductal carcinoma In situ (DCIS) with adequate amounts of surrounding normal breast tissue may prevent the local recurrence of the DCIS. Radiation to the breast and/or tamoxifen may reduce the chance of DCIS occurring in the remaining breast tissue. This is important because DCIS, If left untreated, may develop into invasive breast cancer. Nevertheless, there are serious side effects or sequelae to these treatments. There is, therefore, a need for efficacious breast cancer treatments.

3 00 O There were an estimated 28,300 new cases of pancreatic cancer in the United States in 2000. Over the past 20 years, rates Sof pancreatic cancer have declined in men. Rates among women have remained approximately constant but may be S beginning to decline. Pancreatic cancer caused an estimated 28, 200 deaths in 2000 in the United States. Over the past Syears, there has been a slight but significant decrease in mortality rates among men (about-0. 9% per year) while rates have Sincreased slightly among women.

Surgery, radiation therapy, and chemotherapy are treatment options for pancreatic cancer. These treatment options can extend survival and/or relieve symptoms in many patients but are not likely to produce a cure for most. There is a Ssignificant need for additional therapeutic and diagnostic options for pancreatic cancer.

SUMMARY OF THE INVENTION 00 The present invention relates to a gene, designated 109P1D4, that has now been found to be over-expressed in the cancer(s) listed in Table I. Northern blot expression analysis of 109P1D4 gene expression in normal tissues shows a C( restricted expression pattem in adult tissues. The nucleotide (Figure 2) and amino acid (Figure 2, and Figure 3) sequences of 109P1D4 are provided. The tissue-related profile of 109P1D4 in normal adult tissues, combined with the over-expression observed in the tissues listed in Table I, shows that 109P104 is aberrant over-expressed in at least some cancers, and thus serves as a useful diagnostic, prophylactic, prognostic, and/or therapeutic target for cancers of the tissue(s) such as those listed in Table I.

The invention provides polynucleotides corresponding or complementary to all or part of the 109P1D4 genes, mRNAs, and/or coding sequences, preferably in isolated form, including polynucleotides encoding 109P1D4-related proteins and fragments of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or more than 25 contiguous amino acids at least 30, 35, 40, 45, 50, 55, 60, 65, 70, 80, 85, 90, 95, 100 or more than 100 contiguous amino acids of a 109P1D4-related protein, as well as the peptides/proteins themselves; DNA, RNA, DNAIRNA hybrids, and related molecules, polynucleotides or oligonucleotides complementary or having at least a 90% homology to the 109P1D4 genes or mRNA sequences or parts thereof, and polynucleotides or oligonucleotides that hybridize to the 109P1D4 genes, mRNAs, or to 109P1D4-encoding polynucleotides. Also provided are means for isolating cDNAs and the genes encoding 109P1D4.

Recombinant DNA molecules containing 109P1D4 polynucleotides, cells transformed or transduced with such molecules, and host-vector systems for the expression of 109P1D4 gene products are also provided. The invention further provides antibodies that bind to 109P1 D4 proteins and polypeptide fragments thereof, including polyclonal and monoclonal antibodies, murine and other mammalian antibodies, chimeric antibodies, humanized and fully human antibodies, and antibodies labeled with a detectable marker or therapeutic agent. in certain embodiments, there is a proviso that the entire nucleic acid sequence of Figure 2 is not encoded and/or the entire amino acid sequence of Figure 2 is not prepared. In certain embodiments, the entire nucleic acid sequence of Figure 2 is encoded and/or the entire amino acid sequence of Figure 2 is prepared, either of which are in respective human unit dose forms.

One embodiment of the invention comprises an isolated polynucleotide that encodes a 109P1 D4 protein, wherein the polynucleotide is selected from the group consisting of: a polynucleotide comprising the sequence of SEQ ID NO:2, from nucleotide residue numbers 846 through 3911; a polynucleotide comprising the sequence of SEQ ID NO:4, from nucleotide residue numbers 503 through 3667; 00

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(N a polynucleotide comprising the sequence of SEQ ID NO:6, from nucleotide residue numbers 846 Sthrough 4889; C) a polynucleotide comprising the sequence of SEQ ID NO:8, from nucleotide residue numbers 846 l. through 4778;

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a polynucleotide comprising the sequence of SEQ ID NO:10, from nucleotide residue numbers 846 through 4778; a polynucleotide comprising the sequence of SEQ ID NO:12, from nucleotide residue numbers 614 Sthrough 3727; a polynucleotide comprising the sequence of SEQ ID NO:14, from nucleotide residue numbers 735 00 through 3881; a polynucleotide comprising the sequence of SEQ ID NO:16, from nucleotide residue numbers 735

C

N through 4757; a polynucleotide comprising the sequence of SEQ ID NO:18, from nucleotide residue numbers 514 through 3627; and a polynucleotide of any one of wherein T can also be U.

00 The invention further provides methods for detecting the presence and status of 109P1D4 polynucleotides and proteins in various biological samples, as well as methods for identifying cells that express 109P1D4. A typical embodiment U) of this invention provides methods for monitoring 109P1D4 gene products in a tissue or hematology sample having or V/j) suspected of having some form of growth dysregulation such as cancer.

SThe invention further provides various immunogenic or therapeutic compositions and strategies for treating cancers that express 109P1D4 such as cancers of tissues listed in Table 1, including therapies aimed at inhibiting the transcription, 0 translation, processing or function of 109P1D4 as well as cancer vaccines. In one aspect, the invention provides Scompositions, and methods comprising them, for treating a cancer that expresses 109P1D4 in a human subject wherein the composition comprises a carrier suitable for human use and a human unit dose of one or more than one agent that inhibits 00 the production or function of 109P1D4. Preferably, the carrier is a uniquely human carrier. In another aspect of the invention, C the agent is a moiety that is immunoreactive with 109P1D4 protein. Non-limiting examples of such moieties include, but are C( not limited to, antibodies (such as single chain, monoclonal, polyclonal, humanized, chimeric, or human antibodies), functional equivalents thereof (whether naturally occurring or synthetic), and combinations thereof. The antibodies can be conjugated to a diagnostic or therapeutic moiety. In another aspect, the agent is a small molecule as defined herein.

In another aspect, the agent comprises one or more than one peptide which comprises a cytotoxic T lymphocyte (CTL) epitope that binds an HLA class I molecule in a human to elicit a CTL response to 109P1D4 and/or one or more than one peptide which comprises a helper T lymphocyte (HTL) epitope which binds an HLA class 11 molecule in a human to elicit an HTL response. The peptides of the invention may be on the same or on one or more separate polypeptide molecules. In a further aspect of the invention, the agent comprises one or more than one nucleic acid molecule that expresses one or more than one of the CTL or HTL response stimulating peptides as described above. In yet another aspect of the invention, the one or more than one nucleic acid molecule may express a moiety that is immunologically reactive with 109P1D4 as described above. The one or more than one nucleic acid molecule may also be, or encodes, a molecule that inhibits production of 109P1D4. Non-limiting examples of such molecules include, but are not limited to, those complementary to a nucleotide sequence essential for production of 109P1D4 antisense sequences or molecules that form a triple helix with a nucleotide double helix essential for 109P1D4 production) or a ribozyme effective to lyse 109P1D4 mRNA.

Note that to determine the starting position of any peptide set forth in Tables VIII-XXI and XXII to XLIX (collectively HLA Peptide Tables) respective to its parental protein, variant 1, variant 2, etc., reference is made to three factors: the particular variant, the length of the peptide in an HLA Peptide Table, and the Search Peptides in Table VII. Generally, a unique Search Peptide is used to obtain HLA peptides of a particular for a particular variant. The position of each Search Peptide relative to its respective parent molecule is listed in Table Vil. Accordingly, if a Search Peptide begins at position one must add the value"X-1"to each position in Tables VIII-XXI and XXII to XLIX to obtain the actual position of the HLA peptides in their parental molecule. For example, if a particular Search Peptide begins at position 150 of its parental molecule, one must add 150-1, i. 149 to each HLA peptide amino acid position to calculate the position of that amino acid in the parent molecule.

One embodiment of the invention comprises an HLA peptide, that occurs at least twice in Tables VIII-XXI and XXII to XLIX collectively, or an oligonucleotide that encodes the HLA peptide. Another embodiment of the invention comprises an HLA peptide that occurs at least once in Tables VIII-XXI and at least once in tables XXII to XLIX, or an oligonucleotide that encodes the HLA peptide.

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(Ni Another embodiment of the invention comprises an isolated 109P1D4 protein, wherein the 109P1D4 protein S comprises a polypeptide sequence as set forth in any one of SEQ ID NOs: 3, 5, 7, 9,11, 13,15,17 or 19.

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)Another embodiment of the invention comprises an isolated antibody or fragment thereof that immunospecifically i) binds to an epitope on a 109P1D4 protein, wherein the 109P1D4 protein comprises a polypeptide sequence as set forth in any one of SEQ ID NOs: 3, 5, 7, 9, 11, 13, 15, 17 or 19.

Another embodiment of the invention is antibody epitopes, which comprise a peptide regions, or an oligonucleotide C encoding the peptide region, that has one two, three, four, or five of the following characteristics: 00 6 00 i) a peptide region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to the full length of that protein in Figure 3, that indudes an amino add position having a value equal to or greater than S0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Hydrophilicity profile of Figure II) a peptide region of at least 5 amino adds of a particular peptide of Figure 3, In any whole number increment up C/J to the full length of that protein in Figure 3, that indudes an amino acid position having a value equal to or less than 0.5, 0.4, V' 0.3, 0.2, 0.1, or having a value equal to 0.0, in the Hydropathicity profile of Figure 6; iii) a peptide region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to the full length of that protein in Figure 3, that includes an amino add position having a value equal to or greater than S0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Percent Accessible Residues profile of Figure 7; iv) a peptide region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to the full length of that protein In Figure 3, that includes an amino add position having a value equal to or greater than 0 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Average Rexibility profile of Figure 8; or 0v) a peptide region of at least 5 amino adds of a particular peptide of Figure 3, in any whole number increment up Sto the full length of that protein in Figure 3, that includes an amino add position having a value equal to or greater than 0.6, 0.7,0.8, 0.9, or having a value equal to 1.0, in the Beta-turn profile of Rgure 9.

BRIEF DESCRIPTION OF THE FIGURES Figure 1. The 109P1D4 SSH sequence of 192 nudeotides.

Figure 2. A) The cDNA and amino add sequence of 109P1D4 variant 1 (also called 109P1D4 v.1" or 109P1 D4 variant is shown in Figure 2A. The start methionine is underlined. The open reading frame extends from nuceic acid 846-3911 including the stop codon.

B) The cDNA and amino acd sequence of 109P1D4 variant 2 (also called '109P1D4 is shown in Figure 2B.

The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 503-3667 induding the stop codon.

C) The cDNA and amino acid sequence of 109P1D4 variant 3 (also called '109P1D4 is shown in Figure 2C.

The codon for the start methionine is underlined. The open reading frame extends from nudeic acid 846-4889 including the stop codon.

D) The cDNA and amino add sequence of 109P1D4 variant 4 (also called "109P1D4 v.4) Is shown in Figure 2D.

The codon for the start methionine is underlined. The open reading frame extends from nudeic add 846-4859 including the stop codon.

E) The cDNA and amino acid sequence of 109P1D4 variant 5 (also called "109P1D4 is shown in Figure 2E.

The codon for the start methionine is underlined. The open reading frame extends from nucleic acd 846-4778 including the stop codon.

F) The cDNA and amino add sequence of 109P1D4 variant 6 (also called '109P1D4 Is shown in Figure 2F.

The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 614-3727 including the stop codon.

G) The cDNA and amino acid sequence of 109P1 D4 variant 7 (also called 109P1D4 v.7) Is shown in Figure 2G.

The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 735-3881 including the stop codon.

H) The cDNA and amino add sequence of 109P1D4 variant 8 (also called "109P1D4 v.8) is shown in Figure 2H.

The codon for the start methionine Is underlined. The open reading frame extends from nucleic acid 735-4757 including the stop codon.

00 1) The cDNA and amino acid sequence of 109P1D4 variant 9 (also called "109P1D4 v.9) is shown in Figure 21.

The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 514-3627 including the C stop codon.

r J) 109PID4 v.1, v.2 and v.3 SNP variants. Though these SNP variants are shown separately, they can also CJ/ occur In any combinations and In any of the transcript variants listed above.

V' K) 109P1D4v.6, v.7 and v.8 SNP variants. Though these SNP variants are shown separately, they can also occur in at combinations and in any of the transcript variants listed above.

Figure 3.

SA) The amino acd sequence of 109P1D4 v.1 is shown in Figure 3A; it has 1021 amino acids.

B) The amino add sequence of 109P1 D4 v.2 is shown in Figure 38; it has 1054 amino acids.

C) The amino acid sequence of 109P1D4 v.3 Is shown In Figure 3C; it has 1347 amino ads.

DC) The amino acid sequence of 109P1D4 v.4 is shown in Figure 3D; it has 1337 amino adds.

The amino ad sequence of 109P1D4 v. is shown in Figure 3; it has 13 amino ads.

SE) The amino acid sequence of 109P1D4 v.5 is shown in Figure 3E; it has 1310 amino acids.

SF) The amino acid sequence of 109P1D4 v.6 is shown in Figure 3F; it has 1037 amino acids.

G) The amino acid sequence of 109P1D4 v.7 is shown in Figure 3G; it has 1048 amino acids.

H) The amino add sequence of 109P1D4 v.8 is shown in Figure 3H; it has 1340 amino acids.

I) The amino acid sequence of 109P1D4 v.9 is shown In Figure 31; it has 1037 amino acids.

As used herein, a reference to 109P1D4 Includes all variants thereof, Including those shown in Figures 2, 3, 10, 11, and 12 unless the context clearly indicates otherwise.

Figure 4. Alignment of 109P1 D4 v.1 Protein with protocadherin-11.

Figure 5. Hydrophilicity amino add profile of 109P1D4 v.1-v.9 determined by computer algorithm sequence analysis using the method of Hopp and Woods (Hopp Woods 1981. Proc. Natl. Acad. Sd. U.S.A. 78:3824-3828) accessed on the Protscale website located on the World Wide Web at (expasy.chicgi-bin/protscale.pl) through the ExPasy molecular biology server.

Figure 6. Hydropathidty amino add profile of 109P1D4 v.1-v.9 determined by computer algorithm sequence analysis using the method of Kyte and Doolittle (Kyte Doolittle 1982. J. Mol. Biol. 157:105-132) accessed on the ProtScale website located on the World Wide Web at (.expasy.ch/cgi-bin/protscale.pl) through the ExPasy molecular biology server.

Figure 7. Percent accessible residues amino acid profile of 109P1 D4 v.1-v.9 determined by computer algorithm sequence analysis using the method of Janin (Janin 1979 Nature 277:491-492) accessed on the ProtScale website located on the World Wide Web at (.expasy.ch/cgi-bin/protscale.pl) through the ExPasy molecular biology server.

Figure 8. Average flexibility amino acid profile of 109P1D4 v.1-v.9 determined by computer algorithm sequence analysis using the method of Bhaskaran and Ponnuswamy (Bhaskaran and Ponnuswamy 1988. Int J. Pept Protein Res. 32:242-255) accessed on the ProtScale website located on the World Wide Web at (.expasy.ch/ogi-bin/protscale.pl) through the ExPasy molecular biology server.

Figure 9. Beta-tum amino acid profile of 109P1D4 v.1-v.9 determined by computer algorithm sequence analysis using the method of Deleage and Roux (Deleage, Roux B. 1987 Protein Engineering 1:289-294) accessed on the ProtScale website located on the World Wide Web at (.expasy.ch/cg-bin/protscale.pl) through the ExPasy molecular biology server.

Figure 10. Structure of transcript variants of 109PiD4. Variants 109P1D4 v.2 through v.9 were transcript variants of v.1. Variant v.2 shared middle portion of v.1 sequence (the 3' portion of exon 1 and 5' portion of exon Variant 00 v.6 was similar to v.2 but added an extra exon between exons 1 and 2 of v.2. V.3 shared exon 1 and 5' portion of exon 2 with v.1 with five additional exons downstream. Compared with v.3, v.4 deleted exon 4 of v.3 while v.5 deleted exons 3 and 4 of LC v.3. Variant v.5 lacked exons 3 and 4. This gene (called PCD11) is located in sex chromosomes X and Y. Ends of exons in Sthe transcripts are marked above the boxes. Potential exons of this gene are shown in order as on the human genome. Poly SA tails and single nucleotide differences are not shown in the figure. Lengths of introns and exons are not proportional.

n Figure 11. Schematic alignment of protein variants of 109P1D4. Variants 109P1D4 v.2 through v.9 were proteins translated from the corresponding transcript variants. All these protein variants shared a common portion of the sequence, 3-1011 of v.1, except for a few amino acids different in this segment resulted from SNP in the transcripts.

Ci Variant v.6 and v.9 were the same except for two amino acids at 906 and 1001. Variant v.8 was almost the same as except for the N-terminal end, and a 2-aa deletion at 1117-8. Single amino add difference was not shown. Numbers in parentheses corresponded to positions in variant v.3.

C Figure 12. Intentionally Omitted.

00 0 Figure 13. Figures Secondary structure and transmembrane domains prediction for 109P1D4 protein C variants 1-9 (v.1 (SEQ ID NO: 31); v.2- (SEQ ID NO: 32); v.3 (SEQ ID NO: 33); v.4 (SEQ ID NO: 34); v.5- (SEQ ID NO: 35); v.6- (SEQ ID NO: 36); v.7 (SEQ ID NO: 37); v.8 (SEQ ID NO: 38); v.9 (SEQ ID NO: The secondary structures of 109P1D4 protein variants were predicted using the HNN Hierarchical Neural Network method (NPS@: Network Protein Sequence Analysis TIBS 2000 March Vol. 25, No 3 [291]:147-150 Combet Blanchet Geoujon C. and Del6age httpl/pbl.lbcp.fr/cgi-bin/npsa_automat.pl?page-npsa_nn.html), accessed from the ExPasy molecular biology server located on the World Wide Web at (.expasy.ch/tools/). This method predicts the presence and location of alpha helices, extended strands, and random coils from the primary protein sequence. The percent of the protein variant in a given secondary structure is also listed. Figures top panels: Schematic representation of the probability of existence of transmembrane regions of 109P1D4 variants based on the TMpred algorithm of Hofmann and Stoffel which utilizes TMBASE Hofmann, W. Stoffel. TMBASE A database of membrane spanning protein segments Biol. Chem. Hoppe-Seyler 374:166, 1993). Figures bottom panels: Schematic representation of the probability of the existence of transmembrane regions of 109P1 D4 variants based on the TMHMM algorithm of Sonnhammer, von Heijne, and Krogh (Erik L.L Sonnhammer, Gunnar von Heijne, and Anders Krogh: A hidden Markov model for predicting transmembrane helices in protein sequences. In Proc. of Sixth Int Conf. on Intelligent Systems for Molecular Biology, p 175-182 Ed J. Glasgow, T.

Littlejohn, F. Major, R. Lathrop, D. Sankoff, and C. Sensen Menlo Park, CA: AAAI Press, 1998). The TMpred and TMHMM algorithms are accessed from the ExPasy molecular biology server located on the World Wide Web at (.expasy.ch/tools).

Figure 14. Expression of 109P1D4 in Lymphoma Cancer Patient Specimens. RNA was extracted from peripheral blood lymphocytes, cord blood Isolated from normal individuals, and from lymphoma patient cancer specimens.

Northern blots with 10pg of total RNA were probed with the 109P1D4 sequence. Size standards in kilobases are on the side. Results show expression of 109P1 D4 in lymphoma patient specimens but not in the normal blood cells tested.

Figure 15. Expression of 109P1D4 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool, and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 109P1 D4, was performed at 30 cycles of amplification. Results show strong expression of 109P1D4 in all cancer pools tested. Very low expression was detected in the vital pools.

Figure 16. Expression of 109PID4 In normal tissues. Two multiple tissue northern blots (Clontech), both with 2 pg of mRNA/lane, were probed with the 109P1 D4 SSH fragment Size standards in kilobases (kb) are indicated on the side.

00 Results show expression of approximately 10 kb 109P1D4 transcript in ovary. Weak expression was also detected in placenta and brain, but not in the other normal tissues tested.

C Figure 17. Expression of 109P1 D4 In human cancer cell lines. RNA was extracted from a number of human Sprostate and bone cancer cell lines. Northem blots with 10 pg of total RNAlane were probed with the 109P1D4 SSH fragment Size standards in kilobases (kb) are indicated on the side. Results show expression of 109P1D4 in LAPC-9AD, V LAPC-9AI, LNCaP prostate cancer cell lines, and in the bone cancer cell lines, SK-ES-1 and RD-ES.

Figure 18. Figure 18A: 109P1D4 Expression in Human Normal Tissues. An cDNA dot blot containing 76 different samples from human tissues was analyzed using a 109P1D4 SSH probe. Expression was only detected in multiple Sareas of the brain, placenta, ovary, and fetal brain, amongst all tissues tested. Figure 18B: Expression of 109P1D4 In 0 patient cancer specimens. Expression of 109P1D4 was assayed In a panel of human cancers and their respective matched normal tissues on RNA dot blots. Upregulated expression of 109P1D4 in tumors compared to normal tissues S was observed in uterus, lung and stomach. The expression detected In normal adjacent tissues (isolated from diseased 0 tissues) but not in normal tissues (isolated from healthy donors) may Indicate that these tissues are not fully normal and that 109P1D4 may be expressed in early stage tumors.

Figure 19. 109P1D4 Expression In Lung Cancer Patient Specimens. RNA was extracted from normal lung, prostate cancer xenograft LAPC-9AD, bone cancer cell line RD-ES, and lung cancer patient tumors. Northern blots with pg of total RNA were probed with 109P1D4. Size standards in kilobases are on the side. Results show strong expression of 109P1 D4 in lung tumor tissues as well as the RD-ES cell line, but not in normal lung.

Figure 20. Expression of soluble secreted Tag5 109P1D4 in 293T cells. 293T cells were transfected with either an empty vector or with the Tag5 secretion vector encoding the extracellular domain (ECD; amino acids 24-812) of 109P1D4 variant 1 fused to a Myc/His epitope Tag. 2 days later, cells and media harvested and analyzed for expression of the recombinant Tag5 109P1D4 protein by SDS-PAGE followed by anti-His epitope tag Western blotting. An arrow Indicates the immunoreactive band corresponding to the 109P1D4 ECD present in the media and the lysate from Tag5 109P1D4 transfected cells.

Figure 21. Expression of 109P1 D4 protein In 293T cells. 293T cells were transfected with either an empty vector or with pCDNA3.1 vector encoding the full length cDNA of 109P1D4 variant 1 fused to a Myc/His epitope Tag. 2 days later, cells were harvested and analyzed for expression of 109P1D4 variant 1 protein by SDS-PAGE followed by anti-His epitope tag Western blotting. An arrow indicates the immunoreactive band corresponding to the full length 109P1D4 variant 1 protein expressed in cells transfected with the 109P1 D4 vector but not In control cells.

Figure 22. Tyrosine phosphorylation of 109P1 D4 after pervanadate treatment 293T cells were transfected with the neomycin resistance gene alone or with 109P1D4 In pSRp vector. Twenty four hours after transfection, the cells were either left in 10% serum or grown in 0.1% serum ovemight The cells were then left untreated or were treated with 200 pM pervanadate (1:1 mixture of Na3V04 and H202) for 30 minutes. The cells were lysed in Triton X-100, and the 109P1 D4 protein was Immunoprecipitated with ant-His monoclonal antibody. The Immunoprecipltates were run on SDS-PAGE and then Western blotted with either antiphosphotyrosine (upper panel) or anti-His (lower panel). The 109P D4 protein is phosphorylated on tyrosine In response to pervanadate treatment, and a large amount of the protein moves to the Insoluble fraction following pervanadate-lnduced activation.

Figure 23. Effect of 109P1D4 RNA on cell proliferation. LNCaP cells were transfected with Upofectamine 2000 alone or with siRNA oligonudeotides. The slRNA oligonucleotides included a negative control, Luc4, specific for Luciferase, a positive control, Eg5, specific for the mitotic spindle protein Eg5, or three siRNAs specific for the 109P1D4 protein, 109P1D4.a, 109P1D4.c and 109P1D4.d at 20 nM concentration. Twenty four hours after transfection, the cells were pulsed 0C) with 3 H--thymidine and incorporation was measured after 72 hours. All three siRNAs to 109PI D4 inhibited the proliferation of LNCaP cells, indicating that 109PI D4 expression Is important for the cell growth pathway of these cancer cells.

DETAILED DESCRIPTION OF THE INVENTION Outline of Sections Definitions I1.) 109P1D4 Polyriuceotidez ll.A. Uses of 109P1D14 Polynucleotides; II.A.1.) Monitoring of Genetic Abnormalities (71 I.A2.) Antlsense Embodiments IIL3.) Primers and Primer Pairs IIA.4.) Isolation of 109P1 04-Encoding Nucleic Acid Molecules 00 Recombinant Nucleic Acid Molecules and Host-Vector Systems NI.) 109P1 04-rellated Proteins (7K1flMA) Motif-bearing Protein Embodiments Expression of 109P1 04-related Proteins III.C.) Modifications of I109P1 D4-related Proteins 11.D.) Uses of 109PI 04-related Proteins IV.) 109P104 Antibodies 109P1 D4 Cellular Immune Responses Vi.) 109PID4 Transgenlc Animals VII.) Methods for the Detection of ID9Pi D4 Vill.) Methods for Monitring the Status of 109P11 04-related Genes and Their Products IX) Identification of Molecules That Interact With 11OSPI D4 Therapeutic Methods and Compositions XA.) Anti-Cancer Vaccines 109P1 D4 as a Target for Antibody-Based Therapy XC.) I09P104 as a Target for Cellular Immune Responses XC.. Minigene Vaccines XC.2. Combinations of CTL Peptides; with Helper Peptides XC.3. Combinations of CTL Peptdes with T Cell Priming Agents XC.4. Vaccine Compositions Comprising DC Pulsed with CfL and/or HTIL Peptides XD.) Adoptive Immunotherapy XE.) Administration of Vaccines for Therapeutic or Prophylactic Purposes Diagnostic and Prognostic Embodiments of I109P1I 04.

X(11.) Inhibition of 109P104 Protein Function X(IIA) Inhibition of 11OOPID4 With Intracellular Anitodies XII.B.) Inhibition of 109P1D4 with Recombinant Proteins XII.C.) Inhibition of 109P1D4 Transcription or Translaton XI.D.) General Considerations for Therapeutic Strategies XIL Identification, Characterization and Use of Modulators of 10913134 XIV.) KITSfArddces of Manufacture 00 1I.)-Definitions: SUnless otherwise defined, all terms of art, notations and other scientific terms or terminology used herein are Sintended to have the meanings commonly understood by those of skill in the art to which this invention pertains. In some V) cases, terms with commonly understood meanings are defined herein for clarity and/or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art. Many of the techniques and procedures described or referenced herein are well understood and Scommonly employed using conventional methodology by those skilled in the art, such as, for example, the widely utilized Smolecular cloning methodologies described in Sambrook et al., Molecular Cloning A Laboratory Manual 2nd. edition (1989) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N. Y. As appropriate, procedures involving the use of 00 commercially available kits and reagents are generally carried out in accordance with manufacturer defined protocols and/or parameters unless otherwise noted.

C Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is solely for the purpose of providing a context for the present invention. It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application.

Throughout this specification the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

The terms "advanced prostate cancer", "locally advanced prostate cancer", "advanced disease" and "locally advanced disease" mean prostate cancers that have extended through the prostate capsule, and are meant to include stage C disease under the American Urological Association (AUA) system, stage CI-C2 disease under the Whitmore-Jewett system, and stage T3-T4 and N+ disease under the TNM (tumor, node, metastasis) system. In general, surgery is not recommended for patients with locally advanced disease, and these patients have substantially less favorable outcomes compared to patients having clinically localized (organ-confined) prostate cancer. Locally advanced disease is clinically identified by palpable evidence of induration beyond the lateral border of the prostate, or asymmetry or induration above the prostate base. Locally advanced prostate cancer is presently diagnosed pathologically following radical prostatectomy if the tumor invades or penetrates the prostatic capsule, extends into the surgical margin, or invades the seminal vesicles.

"Altering the native glycosylation pattem" is intended for purposes herein to mean deleting one or more carbohydrate moieties found in native sequence 109P1 D4 (either by removing the underlying glycosylation site or by deleting the glycosylation by chemical and/or enzymatic means), and/or adding one or more glycosylation sites that are not present in the native sequence 109P1D4. In addition, the phrase includes qualitative changes in the glycosylation of the native proteins, involving a change in the nature and proportions of the various carbohydrate moieties present.

The term "analog" refers to a molecule which is structurally similar or shares similar or corresponding attributes with another molecule g. a 109P1 4-related protein). For example, an analog of a 109P1 4 protein can be specifically bound by an antibody or T cell that specifically binds to 109P1 D4.

The term "antibody" is used in the broadest sense. Therefore, an "antibody" can be naturally occurring or manmade such as monoclonal antibodies produced by conventional hybridoma technology. Anti-109P1D4 antibodies comprise monoclonal and polyclonal antibodies as well as fragments containing the antigen-binding domain and/or one or more complementarity determining regions of these antibodies.

00 O An "antibody fragment" is defined as at least a portion of the variable region of the immunoglobulin molecule that binds to its target, the antigen-binding region. In one embodiment, it specifically covers single anti-109P1D4 antibodies C and clones thereof (including agonist, antagonist and neutralizing antibodies) and anti-109P1D4 antibody compositions with polyepitopic specificity.

The term "codon optimized sequences" refers to nucleotide sequences that have been optimized for a particular host species by replacing any codons having a usage frequency of less than about 20%. Nucleotide sequences that have been optimized for expression in a given host species by elimination of spurious polyadenylation sequences, elimination of i exonlintron splicing signals, elimination of transposon-like repeats and/or optimization of GC content in addition to codon 00

(N

O-

12 00 optimization are referred to herein as an "expression enhanced sequences." A combinatorial library" is a collection of diverse chemical compounds generated by either chemical synthesis or biological synthesis by combining a number of chemical 'building blocks" such as reagents. For example, a linear y combinatorial chemical library, such as a polypeptide mutein) library, is formed by combining a set of chemical building Sblocks called amino acids in every possible way for a given compound length the number of amino acids in a Spolypeptide compound). Numerous chemical compounds are synthesized through such combinatorial mixing of chemical building blocks (Gallop et al., J. Med. Chem. 37(9): 1233-1251 (1994)).

Preparation and screening of combinatorial libraries is well known to those of skill in the art Such combinatorial Schemical libraries Include, but are not limited to, peptide libraries (see, U.S. Patent No. 5,010,175, Furka, Pept Prot.

0 Res. 37:487-493 (1991), Houghton et al., Nature, 354:84-88 (1991)), peptolds (PCT Publication No WO 91/19735), encoded peptides (PCT Publication WO 93/20242), random blo- oligomers (PCT Publication WO 92/00091), benzodiazepines (U.S.

00 Pat No. 5,288,514), diversomers such as hydantoins, benzodiazepines and dipeptides (Hobbs et al., Proc. Nat Acad. Sd.

SUSA 90:6909-6913 (1993)), vinylogous polypeptides (Hagihara et al., J. Amer. Chem. Soc. 114:6568 (1992)), nonpeptidal Speptidomimetics with a Beta-D-Glucose scaffolding (Hirschmann et al., J. Amer. Chem. Soc. 114:9217-9218 (1992)), analogous organic syntheses of small compound libraries (Chen et al., J. Amer. Chem. Soc. 116:2661 (1994)), oligocarbamates (Cho, et al., Science 261:1303 (1993)), and/or peptidyl phosphonates (Campbell et al., J. Org. Chem.

59:658 (1994)). See, generally, Gordon et al., J. Med. Chem. 37:1385 (1994), nucleic acid libraries (see, Stratagene, Corp.), peptide nucleic acid libraries (see, U.S. Patent 5,539,083), antibody libraries (see, Vaughn et al., Nature Biotechnology 14(3): 309-314 (1996), and PCT/US96/10287), carbohydrate libraries (see, Lang et al., Science 274:1520-1522 (1996), and U.S. Patent No. 5,593,853), and small organic molecule libraries (see, benzodiazepines, Baum, C&EN, Jan 18, page 33 (1993); isoprenoids, U.S. Patent No. 5,569,588; thiazolidinones and metathiazanones, U.S.

Patent No. 5,549,974; pyrrolidines, U.S. Patent Nos. 5,525,735 and 5,519,134; morpholino compounds, U.S. Patent No.

5,506, 337; benzodiazepines, U.S. Patent No. 5,288,514; and the like).

Devices for the preparation of combinatorial libraries are commercially available (see, 357 NIPS, 390 NIPS, Advanced Chem Tech, Louisville KY; Symphony, Rainin, Wobum, MA; 433A, Applied Biosystems, Foster City, CA; 9050, Plus, Millipore, Bedford, NIA). A number of well-known robotic systems have also been developed for solution phase chemistries. These systems include automated workstations such as the automated synthesis apparatus developed by Takeda Chemical Industries, LTD. (Osaka. Japan) and many robotic systems utilizing robotic arms (Zymate H, Zymark Corporation, Hopkinton, Mass.; Orca, Hewlett-Packard, Palo Alto, Calif.), which mimic the manual synthetic operations performed by a chemist Any of the above devices are suitable for use with the present invention. The nature and Implementation of modifications to these devices (if any) so that they can operate as discussed herein will be apparent to persons skilled in the relevant art In addition, numerous combinatorial libraries are themselves commercially available (see, ComGenex, Princeton, NJ; Asinex, Moscow, RU; Tripos, Inc., St. Louis, MO; ChemStar, Ltd, Moscow, RU; 3D Pharmaceuticals, Exton, PA; Martek Biosciences, Columbia, MD; etc.).

The term "cytotoxic agent" refers to a substance that inhibits or prevents the expression activity of cells, function of cells and/or causes destruction of cells. The term is intended to Include radioactive isotopes chemotherapeutic agents, and toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof. Examples of cytotoxic agents include, but are not limited to auristatins, auromycins, maytansinoids, yttrium, bismuth, ridn, ridn A-chain, combrestatin, duocarmycins, dolostatins, doxorubicin, daunorubidn, taxol, cisplatin, cc1065, ethldium bromide, mitomydn, etoposlde, tenoposlde, vincristine, vinblastine, colchicine, dihydroxy anthracin dione, actinomyin, diphtheria toxin, Pseudomonas exotoxin (PE) A, PE40, abrin, abrin A chain, modeccin A chain, 13 00 alpha-sarcin, gelonin, mitogellin, retstrictocin, phenomycin, enomycin, curicin, crotin, calicheamicin, Sapaonaria officinalis inhibitor, and glucocorticoid and other chemotherapeutic agents, as well as radioisotopes such as At 211 1131, 125, Y 9 s, Re 1 6 c Re 18 Sm 1s 3 Bi 212 21 3

P

32 and radioactive isotopes of Lu including Lu 7 n. Antibodies may also be conjugated to an anticancer pro-drug activating enzyme capable of converting the pro-drug to its active form.

C) The "gene product Is sometimes referred to herein as a protein or mRNA. For example, a "gene product of the invention" is sometimes referred to herein as a "cancer amino add sequence', 'cancer protein", "protein of a cancer listed in Table a "cancer mRNA', "mRNA of a cancer fisted in Table etc. In one embodiment, the cancer protein is encoded by a nucleic add of Figure 2. The cancer protein can be a fragment, or alternatively, be the full-length protein to the fragment C encoded by the nucleic acids of Figure 2. In one embodiment, a cancer amino add sequence is used to determine 0 sequence Identity or similarity. In another embodiment, the sequences are naturally occurring allelic variants of a protein encoded by a nucleic acid of Figure 2. In another embodiment the sequences are sequence variants as further described l herein.

00 0"High throughput screening" assays for the presence, absence, quantification, or other properties of particular Snudeic adds or protein products are well known to those of skill in the art Similarly, binding assays and reporter gene assays are similarly well known. Thus, U.S. Patent No. 5,559,410 discloses high throughput screening methods for proteins; U.S. Patent No. 5,585,639 disdoses high throughput screening methods for nudeic add binding in arrays); while U.S. Patent Nos. 5,576,220 and 5,541,061 disclose high throughput methods of screening for ligand/antibody binding.

In addition, high throughput screening systems are commercially available (see, Amersham Biosciences, Piscataway, NJ; Zymark Corp., Hopkinton, MA; Air Technical Industries, Mentor, OH; Beckman Instruments, Inc. Fullerton, CA; Precision Systems, Inc., Natick, MA; etc.). These systems typically automate entire procedures, induding all sample and reagent pipetting, liquid dispensing, timed incubations, and final readings of the microplate in detector(s) appropriate for the assay. These configurable systems provide high throughput and rapid start up as well as a high degree of flexibility and customization. The manufacturers of such systems provide detailed protocols for various high throughput systems. Thus, Zymark Corp. provides technical bulletins describing screening systems for detecting the modulation of gene transcription, ligand binding, and the like.

The term "homolog" refers to a molecule which exhibits homology to another molecule, by for example, having sequences of chemical residues that are the same or similar at corresponding positions.

"Human Leukocyte Antigen" or HLA" Is a human dass I or class II Major Histocompatibility Complex (MHC) protein (see, Stites, et al., IMMUNOLOGY, 8TH ED., Lange Publishing, Los Altos, CA (1994).

The terms 'hybridize", "hybridizing', "hybridizes' and the like, used In the context of polynuceotides, are meant to refer to conventional hybridization conditions, preferably such as hybridization in 50% formamide/6XSSC/0.1 SDS/100 pg/ml ssDNA, in which temperatures for hybridization are above 37 degrees C and temperatures for washing in 0.1XSSC/0.1% SDS are above 55 degrees C.

The phrases "isolated' or "biologically pure' refer to material which is substantially or essentially free from components which normally accompany the material as it is found in its native state. Thus, isolated peptides In accordance with the invention preferably do not contain materials normally associated with the peptides in their in situ environment For example, a polynudeotide is said to be "Isolated" when it is substantially separated from contaminant polynudeotides that correspond or are complementary to genes other than the 109P1 D4 genes or that encode polypeptides other than 109P1 D4 gene product or fragments thereof. A skilled artisan can readily employ nucleic add isolation procedures to obtain an Isolated 109P1 D4 polynudeotide. A protein is said to be Isolated," for example, when physical, mechanical or chemical methods are employed to remove the 109P1 D4 proteins from cellular constituents that are normally assodated with the protein. A skilled artisan can readily 00 employ standard purification methods to obtain an isolated 109P1D4 protein. Alternatively, an isolated protein can be prepared by Schemical means.

The term "mammar refers to any organism classified as a mammal, including mice, rats, rabbits, dogs, cats, cows, horses and humans. In one embodiment of the invention, the mammal is a mouse. In another embodiment of the invention, the Smammal is a human.

The terms "metastatic prostate cancer" and "metastatic disease" mean prostate cancers that have spread to regional lymph nodes or to distant sites, and are meant to include stage D disease under the AUA system and stage TxNxM+ under the TNM system. As is the case with locally advanced prostate cancer, surgery is generally not indicated for patients with metastatic disease, and hormonal (androgen ablation) therapy is a preferred treatment modality. Patients with Smetastatic prostate cancer eventually develop an androgen-refractory state within 12 to 18 months of treatment initiation.

Approximately half of these androgen-refractory patients die within 6 months after developing that status. The most common 00 site for prostate cancer metastasis is bone. Prostate cancer bone metastases are often osteoblastic rather than osteolytic resulting in net bone formation). Bone metastases are found most frequently in the spine, followed by the femur, pelvis, S rib cage, skull and humerus. Other common sites for metastasis include lymph nodes, lung, liver and brain. Metastatic prostate cancer is typically diagnosed by open or laparoscopic pelvic lymphadenectomy, whole body radionudide scans, skeletal radiography, and/or bone lesion biopsy.

The term 'modulator" or test compound" or "drug candidate' or grammatical equivalents as used herein describe any molecule, protein, oligopeptide, small organic molecule, polysaccharide, polynucleotide, etc., to be tested for the capacity to directly or indirectly alter the cancer phenotype or the expression of a cancer sequence, a nucleic add or protein sequences, or effects of cancer sequences signaling, gene expression, protein interaction, etc.) In one aspect, a modulator will neutralize the effect of a cancer protein of the invention. By "neutralize" is meant that an activity of a protein is inhibited or blocked, along with the consequent effect on the cell. In another aspect, a modulator will neutralize the effect of a gene, and its corresponding protein, of the invention by normalizing levels of said protein. In preferred embodiments, modulators alter expression profiles, or expression profile nucleic acids or proteins provided herein, or downstream effector pathways. In one embodiment, the modulator suppresses a cancer phenotype, e.g. to a normal tissue fingerprint In another embodiment a modulator induced a cancer phenotype. Generally, a plurality of assay mixtures is run in parallel with different agent concentrations to obtain a differential response to the various concentrations. Typically, one of these concentrations serves as a negative control, at zero concentration or below the level of detection.

Modulators, drug candidates or test compounds encompass numerous chemical classes, though typically they are organic molecules, preferably small organic compounds having a molecular weight of more than 100 and less than about 2,500 Daltons. Preferred small molecules are less than 2000, or less than 1500 or less than 1000 or less than 500 D.

Candidate agents comprise functional groups necessary for structural Interaction with proteins, particularly hydrogen bonding, and typically Include at least an amine, carbonyl, hydroxyl or carboxyl group, preferably at least two of the functional chemical groups. The candidate agents often comprise cydical carbon or heterocydic structures and/or aromatic or polyaromatic structures substituted with one or more of the above functional groups. Modulators also comprise biomolecules such as peptides, saccharides, fatty adds, steroids, purines, pyrimidines, derivatives, structural analogs or combinations thereof. Particularly preferred are peptides. One class of modulators are peptides, for example of from about five to about amino adds, with from about five to about 20 amino adds being preferred, and from about 7 to about 15 being particularly preferred. Preferably, the cancer modulatory protein is soluble, includes a non-transmembrane region, and/or, has an Nterminal Cys to aid in solubility. In one embodiment, the C-terminus of the fragment Is kept as a free acid and the N-terminus Is a free amine to aid in coupling, to cysteine. In one embodiment, a cancer protein of the invention Is conjugated to an 00 immunogenic agent as discussed herein. In one embodiment, the cancer protein is conjugated to BSA. The peptides of the invention, of preferred lengths, can be linked to each other or to other amino acids to create a longer peptide/protein.

CN The modulatory peptides can be digests of naturally occurring proteins as Is outlined above, random peptides, or 'biased" random peptides. In a preferred embodiment peptide/protein-based modulators are antibodies, and fragments thereof, as 7) defined herein.

SModulators of cancer can also be nucleic acids. Nucleic acid modulating agents can be naturally occurring nucleic acids, random nudeic acids, or "biased" random nucleic acids. For example, digests of prokaryotic or eukaryotic genomes can be used in an approach analogous to that outlined above for proteins.

The term "monodonal antibody" refers to an antibody obtained from a population of substantially homogeneous O antibodies, the antibodies comprising the population are identical except for possible naturally occurring mutations that are present in minor amounts.

C A "motif, as In biological motif of a 109P1D4-related protein, refers to any pattem of amino acids forming part of 0the primary sequence of a protein, that is associated with a particular function protein-protein interaction, protein-DNA interaction, etc) or modification that is phosphorylated, glycosylated or amidated), or localization secretory sequence, nuclear localization sequence, etc.) or a sequence that is correlated with being immunogenic, either humorally or cellularly. A motif can be either contiguous or capable of being aligned to certain positions that are generally correlated with a certain function or property. In the context of HLA motifs, "motif refers to the pattern of residues in a peptide of defined length, usually a peptide of from about 8 to about 13 amino acids for a class I HLA motif and from about 6 to about 25 amino acids for a class II HLA motif, which is recognized by a particular HLA molecule. Peptide motifs for HLA binding are typically different for each protein encoded by each human HLA allele and differ In the pattern of the primary and secondary anchor residues.

A "pharmaceutical excipient" comprises a material such as an adjuvant, a carrier, pH-adjusting and buffering agents, tonicity adjusting agents, wetting agents, preservative, and the like.

'Pharmaceutically acceptable" refers to a non-toxic, inert, and/or composition that is physiologically compatible with humans or other mammals.

The term "polynudeotide" means a polymeric form of nudeotides of at least 10 bases or base pairs in length, either ribonucleotides or deoxynudeotides or a modified form of either type of nucleotide, and is meant to include single and double stranded forms of DNA and/or RNA. In the art, this term if often used interchangeably with "ollgonudeotide'. A polynudeotide can comprise a nucleotide sequence disclosed herein wherein thymldlne as shown for example in Figure 2, can also be uracil this definition pertains to the differences between the chemical structures of DNA and RNA, in particular the observation that one of the four major bases In RNA is uracil instead of thymidine The term "polypeptide" means a polymer of at least about 4, 5, 6, 7, or 8 amino acids. Throughout the specification, standard three letter or single letter designations for amino adds are used. In the art, this term Is often used interchangeably with "peptide" or "protein".

An HLA 'primary anchor residue" is an amino add at a specific position along a peptide sequence which is understood to provide a contact point between the Immunogenic peptide and the HLA molecule. One to three, usually two, primary anchor residues within a peptide of defined length generally defines a "motif for an immunogenic peptide. These residues are understood to fit in close contact with peptide binding groove of an HLA molecule, with their side chains buried in specific pockets of the binding groove. In one embodiment, for example, the primary anchor residues for an HLA class I molecule are located at position 2 (from the amino terminal position) and at the carboxyl terminal position of a 8, 9, 10, 11, or 12 residue peptide epitope in accordance with the invention. Alternatively, in another embodiment, the primary anchor residues of a peptide binds an HLA class II molecule are spaced relative to each other, rather than to the termini of a peptide, where the peptide is generally of at least 9 amino acids in length. The primary anchor positions for each motif and supermotif are set forth in Table IV. For example, analog peptides can be created by altering the presence or absence of particular residues in the primary and/or secondary anchor positions shown in Table IV. Such analogs are used to modulate the binding affinity and/or population coverage of a peptide comprising a particular HLA motif or supermotif.

"Radioisotopes' include, but are not limited to the following (non-limiting exemplary uses are also set forth): Examples of Medical Isotopes: Isotope Actinium-225 (AC-225) Actinium-227 (AC-227) Bismuth-212 (Bi-212) Bismuth-213 (Bi-213) Cadmium-109 (Cd-109) Copper-64 (Cu-64) Copper-67 (Cu-67) Dysprosium-166 (Dy-166) Erblum-169 (Er-169) Europium-152 (Eu-152) Europium-154 (Eu-154) Gadolinium-153 (Gd-153) Gold-198 (Au-198) Holmium-166 (Ho-166) lodine-125 (1-125) Iodine-131 (1-131) Irdium-192 (lr-192) Lutetium-177 (Lu-177) Molybdenum-99 (Mo-99) Osmium-194 (Os-194) Description of use See Thorium-229 (Th-229) Parent of Radium-223 (Ra-223) which is an alpha emitter used to treat metastases in the skeleton resulting from cancer breast and prostate cancers), and cancer radioimmunotherapy See Thorium-228 (Th-228) See Thorium-229 (Th-229) Cancer detection Radiation source for radiotherapy of cancer, for food irradiators, and for sterilization of medical supplies A positron emitter used for cancer therapy and SPECT Imaging Beta/gamma emitter used in cancer radioimmunotherapy and diagnostic studies breast and colon cancers, and lymphoma) Cancer radioimmunotherapy Rheumatoid arthritis treatment, particularly for the small joints associated with fingers and toes Radiation source for food irradiation and for sterilization of medical supplies Radiation source for food Irradiation and for sterilization of medical supplies Osteoporosis detection and nuclear medical quality assurance devices Implant and Intracavity therapy of ovarian, prostate, and brain cancers Multiple myeloma treatment in targeted skeletal therapy, cancer radioimmunotherapy, bone marrow ablation, and rheumatoid arthritis treatment Osteoporosis detection, diagnostic imaging, tracer drugs, brain cancer treatment, radiolabeling, tumor Imaging, mapping of receptors in the brain, interstitial radiation therapy, brachytherapy for treatment of prostate cancer, determination of glomerular filtration rate (GFR), determination of plasma volume, detection of deep vein thrombosis of the legs Thyroid function evaluation, thyroid disease detection, treatment of thyroid cancer as well as other non-malignant thyroid diseases Graves disease, goiters, and hyperthyroidism), treatment of leukemia, lymphoma, and other forms of cancer breast cancer) using radioimmunotherapy Brachytherapy, brain and spinal cord tumor treatment, treatment of blocked arteries arteriosclerosis and restenosis), and Implants for breast and prostate tumors Cancer radiolmmunotherapy and treatment of blocked arteries arteriosclerosis and restenosis) Parent of Technetium-99m (Tc-99m) which is used for imaging the brain, liver, lungs, heart, and other organs. Currently, To-99m is the most widely used radioisotope used for diagnostic imaging of various cancers and diseases involving the brain, heart, liver, lungs; also used in detection of deep vein thrombosis of the legs Cancer radioimmunotherapy 00 00 Palladium-103 (Pd-103) Platinum-1 95m (Pt-195m) Phosphorus-32 (P-32) Phosphorus-33 (P-33) Radium-223 (Ra-223) Rhenium-1 86 (Re-186) Rhenium-188 (Re-188) Rhodium-105 (Rh-105) Samarium-145 (Sm-145) Samarium-153 (Sm-i 53) Scandium-47 (Sc-47) Strontium-89 (Sr89) Technetium-99m (TC99m) Thorium-228 (Th-228) Thorium-229 (Th-229) Thulium-170 (Tm-170) Tin-117m (Sn-i 17m) Tungsten-188 (W-188) Xenon-1 27 (Xe-i 27) Ytterbium-175 (Yb-175) Yttrium-91 (Y-91) Prostate cancer treatment Studies on biodistribution and metabolism of cisplatin, a chemotherapeutic drug Polycythemia rubra vera (blood cell disease) and leukemia treatment bone cancer diagnosis/treatment. colon, pancreatic, and river cancer treatment. radiolabeling nucleic acids for in vitro research, diagnosis of superficial tumors, treatment of blocked arteries arteriosclerosis and restenosis), and intracavity therapy Leukemia treatment, bone disease diagnosis/treatment, radiolabeling, and treatment of blocked arteries arteriosclerosis and restenosis) See Actinium-227 (Ac-227) Bone cancer pain relief, rheumatoid arthritis treatment and diagnosis and treatment of lymphoma and bone, breast colon, and liver cancers using radioimmunotherapy Cancer diagnosis and treatment using radioimmunotherapy, bone cancer pain relief, treatment of rheumatoid arthritis, and treatment of prostate cancer Cancer radioimmunotherapy Ocular cancer treatment Cancer radioimmunotherapy and bone cancer pain relief Cancer radioimmunotherapy and bone cancer pain relief Radiotracer used in brain studies, imaging of adrenal cortex by gamma-scintigraphy, lateral locations of steroid secreting tumors, pancreatic scanning, detection of hyperactive parathyroid glands, measure rate of bile acid loss from the endogenous pool Bone cancer detection and brain scans Bone cancer pain relief, multiple myeloma treatment and osteoblastic therapy See Molybdenum-99 (Mo-99) Parent of Bismuth-212 (Bi-212) which is an alpha emitter used in cancer radioimmunotherapy Parent of Actinium-225 (Ao-225) and grandparent of Bismuth-213 (Bi-2 3) which are alpha emitters used in cancer radiolmmunotherapy Gamma source for blood irradiators, energy source for implanted medical devices Cancer immunotherapy and bone cancer pain relief Parent for Rhenium-188 (Re-188) which is used for cancer diagnosticsltreatment, bone cancer pain relief, rheumatoid arthritis treatment and treatment of blocked arteries arteriosdclerosis and restenosis) Neuroimaging of brain disorders, high resolution SPECT studies, pulmonary function tests, and cerebral blood flow studies Cancer radioimmunotherapy Microseeds obtained from irradiating Yttrium-89 (Y-89) for liver cancer treatment A gamma-emitting label for Yttrium-90 (Y-90) which is used for cancer radioimmunotherapy lymphoma, breast, colon, kidney, lung, ovarian, prostate, pancreatic, and inoperable liver cancers) 00

O

SBy 'randomized' or grammatical equivalents as herein applied to nucleic acids and proteins is meant that each Snucleic acid and peptide consists of essentially random nudeotides and amino acids, respectively. These random peptides S(or nucleic acids, discussed herein) can incorporate any nucleotide or amino add at any position. The synthetic process can O be designed to generate randomized proteins or nucleic adds, to allow the formation of all or most of the possible combinations over the length of the sequence, thus forming a library of randomized candidate bioactive proteinaceous agents.

SIn one embodiment, a library is "fully randomized," with no sequence preferences or constants at any position. In another embodiment, the library is a 'biased random" library. That is, some positions within the sequence either are held Sconstant, or are selected from a limited number of possibilities. For example, the nudeotides or amino acid residues are 00 randomized within a defined class, of hydrophobic amino adds, hydrophilic residues, sterically biased (either small or 0 large) residues, towards the creation of nudeic add binding domains, the creation of cysteines, for cross-linking, prolines for c SH-3 domains, serines, threonines, tyrosines or histidines for phosphorylation sites, etc., or to purines, etc.

A "recombinant" DNA or RNA molecule is a DNA or RNA molecule that has been subjected to molecular manipulation in vitr.

Non-limiting examples of small molecules indude compounds that bind or interact with 109P1 D4, ligands including hormones, neuropeptides, chemokines, odorants, phospholipids, and functional equivalents thereof that bind and preferably inhibit 109P1D4 protein function. Such non-limiting small molecules preferably have a molecular weight of less than about kDa, more preferably below about 9, about 8, about 7, about 6, about 5 or about 4 kDa. In certain embodiments, small molecules physically associate with, or bind, 109P1D4 protein; are not found in naturally occurring metabolic pathways; and/or are more soluble In aqueous than non-aqueous solutions 'Stringency' of hybridization reactions is readily determinable by one of ordinary skill in the art, and generally is an empirical calculation dependent upon probe length, washing temperature, and salt concentration. In general, longer probes require higher temperatures for proper annealing, while shorter probes need lower temperatures. Hybridization generally depends on the ability of denatured nucleic acid sequences to reanneal when complementary strands are present in an environment below their melting temperature. The higher the degree of desired homology between the probe and hybridizable sequence, the higher the relative temperature that can be used. As a result, it follows that higher relative temperatures would tend to make the reaction conditions more stringent, while lower temperatures less so. For additional details and explanation of stringency of hybridization reactions, see Ausubel et al., Current Protocols in Molecular Biology, Wiley Interscence Publishers, (1995).

'Stringent conditions" or "high stringency conditions', as defined herein, are identified by, but not limited to, those that employ low ionic strength and high temperature for washing, for example 0.015 M sodium chloride/0.0015 M sodium citrate/0.1% sodium dodecyl sulfate at 50oC; employ during hybridization a denaturing agent, such as formamide, for example, 50% formamide with 0.1% bovine serum albumin/0.1% Ficoll/0.1% polyvinylpyrrolidone/50 mM sodium phosphate buffer at pH 6.5 with 750 mM sodium chloride, 75 mM sodium citrate at 42 oC; or employ 50% formamide, 5 x SSC (0.75 M NaCI, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 0.1% sodium pyrophosphate, 5 x Denhardfs solution, sonicated salmon sperm DNA (50 lpg/ml), 0.1% SDS, and 10% dextran sulfate at 42 oC, with washes at 420C in 0.2 x SSC (sodium chloride/sodium, citrate) and 50% formamide at 55 oC, followed by a high-stringency wash consisting of 0.1 x SSC containing EDTA at 55 oC. "Moderately stringent conditions' are described by, but not limited to, those in Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Press, 1989, and Include 19 00 the use of washing solution and hybridization conditions temperature, ionic strength and %SDS) less stringent than those described above. An example of moderately stringent conditions is overnight incubation at 370C in a solution C comprising: 20% formamide, 5 x SSC (150 mM NaCI, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 5 x Denhardrs solution, 10% dextran sulfate, and 20 mg/mL denatured sheared salmon sperm DNA, followed by washing the Cfilters in 1 x SSC at about 37-50oC. The skilled artisan will recognize how to adjust the temperature, ionic strength, etc. as necessary to accommodate factors such as probe length and the like.

An HLA "supermotif" is a peptide binding specificity shared by HLA molecules encoded by two or more HLA alleles.

Overall phenotypic frequencies of HLA-supertypes in different ethnic populations are set forth in Table IV The non- Slimiting constituents of various supetypes are as follows: A2: A*0201, A'0202, A*0203, A'0204, A* 0205, A*0206, A*6802, A*6901, A*0207 A3, A3, All, A31, A'3301, A'6801, A*0301, A*1101, A*3101 C B7: 87, 8*3501-03, B*51, B*5301, B*5401, B*5501, B*5502, B*5601, 8*6701, B'7801, B*0702, B*5101, B*5602 00 0B44: B*3701, B*4402, B'4403, B*60 (B*4001), 861 (B'4006) A1 A*0102, A*2604, A*3601, A'4301, A'8001 A24: A*24, A*30, A*2403, A*2404, A'3002, A*3003 B27 B*1401-02, B*1503, B*1509, B*1510, B*1518, B*3801-02, B*3901, B*3902, B*3903-04, B*4801-02, 8B7301, B*2701-08 58: 8*1516, B*1517, B*5701, B*5702, B58 862:_B*4601, B52, B*1501 (B62), B*1502 (B75), B*1513 (877) Calculated population coverage afforded by different HLA-supertype combinations are set forth in Table IV As used herein "to treat or 'therapeutic" and grammatically related terms, refer to any improvement of any consequence of disease, such as prolonged survival, less morbidity, and/or a lessening of side effects which are the byproducts of an alternative therapeutic modality; full eradication of disease is not required.

A'transgenic animal" a mouse or rat) is an animal having cells that contain a transgene, which transgene was introduced into the animal or an ancestor of the animal at a prenatal, an embryonic stage. A transgene" is a DNA that is integrated into the genome of a cell from which a transgenic animal develops.

As used herein, an HLA or cellular immune response "vaccine" is a composition that contains or encodes one or more peptides of the invention. There are numerous embodiments of such vaccines, such as a cocktail of one or more individual peptides; one or more peptides of the invention comprised by a polyepitopic peptide; or nudeic acids that encode such individual peptides or polypeptides, a minigene that encodes a polyepitopic peptide. The "one or more peptides" can include any whole unit integer from 1-150 or more, at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39,40,41, 42,43, 44, 45, 46,47,48, 49, 50, 65,70, 75, 80, 85, 90, 95, 100, 105, 110,115, 120,125,130,135,140,145, or 150 or more peptides of the invention.

The peptides or polypeptides can optionally be modified, such as by lipidation, addition of targeting or other sequences. HLA dass I peptides of the invention can be admixed with, or linked to, HLA class II peptides, to facilitate activation of both cytotoxlc T lymphocytes and helper T lymphocytes. HLA vaccines can also comprise peptide-pulsed antigen presenting cells, dendritic cells.

The term "variant" refers to a molecule that exhibits a variation from a described type or norm, such as a protein that has one or more different amino acd residues in the coresponding position(s) of a specifically described protein the 109P1D4 protein shown in Figure 2 or Figure 3. An analog Is an example of a variant protein. Splice Isoforms and single nudeotides polymorphisms (SNPs) are further examples of variants.

00 The "109P104-related proteins" of the invention include those specifically identified herein, as well as allelic variants, conservative substitution variants, analogs and homologs that can be isolated/generated and characterized without undue experimentation following the methods outlined herein or readily available in the art Fusion proteins that combine parts of different 109P1D4 proteins or fragments thereof, as well as fusion proteins of a 109P1D4 protein and a heterologous polypeptide are also induded. Such 109P1D4 proteins are collectively referred to as the 109P1D4-related proteins, the proteins of the invention, or 109P1D4. The term *109P1D4-related protein" refers to a polypeptide fragment or a 109P1D4 protein sequence of 4, 5,6,7, 8,9,10,11,12,13,14,15,16,17,18,19, 20, 21, 22, 23, 24,25, or more than 25 amino acids; or, at least 30, 35, 40, 55, 60, 65, 70, 80, 85, 90, 95, 100, 105, 110,115,120,125,130,135,140,145,150,155, 160, 165,170,175,180,185, 190,195, 200, 225,250, 275, 300, 325, 350, 375, 400, 425,450, 475, 500, 525, 550, 575, or 576 or more amino adds.

II.) 109P1D4 Polynucleotides One aspect of the invention provides polynudeotides corresponding or complementary to all or part of a 109P1 D4 00 gene, mRNA, and/or coding sequence, preferably in isolated form, including polynudeotides encoding a 109P1D4-related protein and fragments thereof, DNA, RNA, DNA/RNA hybrid, and related molecules, polynucleotides or oligonucleotides CKl complementary to a 109P1 D4 gene or mRNA sequence or a part thereof, and polynucleotides or oligonucleoUdes that hybridize to a 109P1D4 gene, mRNA, or to a 109P1D4 encoding polynucleotide (collectively, *109P1D4 polynucleotides). In all Instances when referred to in this section, T can also be U in Figure 2.

Embodiments of a 109P1 D4 polynucleotide Include: a 109P1 D4 polynucleotide having the sequence shown in Figure 2, the nucleotide sequence of 109P1D4 as shown in Figure 2 wherein T is U; at least 10 contiguous nucleotides of a polynudeotide having the sequence as shown in Figure 2; or, at least 10 contiguous nucleotides of a polynucleotide having the sequence as shown in Figure 2 where T is U. For example, embodiments of 109P1 D4 nucleotides comprise, without limitation: a polynudeotide comprising, consisting essentially of, or consisting of a sequence as shown in Figure 2, wherein T can also be U; (II) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in Figure 2A, from nucleotide residue number 846 through nucleotide residue number 3911, induding the stop codon, wherein T can also be U; (111) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in Figure 2B, from nucleotide residue number 503 through nucleotide residue number 3667, including the stop codon, wherein T can also be U; (IV) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in Figure 2C, from nudeotide residue number 846 through nudeotide residue number 4889, including the a stop codon, wherein T can also be U; a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in Figure 2D, from nucleotide residue number 846 through nucleotide residue number 4859, Including the stop codon, wherein T can also be U; (VI) a polynudeotde comprising, consisting essentially of, or consisting of the sequence as shown in Figure 2E, from nucleotide residue number 846 through nucleotide residue number 4778, Including the stop codon, wherein T can also be U; 21 C0 (VII) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in Figure 2F, from nudeotide residue number 614 through nudeotide residue number 3727, including the stop codon, CKl wherein T can also be U; U) (VIII) a polynudeotide comprising, consisting essentially of, or consisting of the sequence as shown In Figure S2G, from nudeotide residue number 735 through nucleotide residue number 3881, including the stop codon, wherein T can also be U; (IX) a polynuceotide comprising, consisting essentially of, or consisting of the sequence as shown in Figure 2H, from nudeotide residue number 735 through nudeotide residue number 4757, induding the stop codon, wherein T can also be U; a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in Figure N 21, from nudeotide residue number 514 through nucleotide residue number 3627, including the stop codon, 00 wherein T can also be U; C (XI) a polynuleotide that encodes a 109P1 D4-related protein that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100% homologous to an entire amino acid sequence shown in Figure 2A-I; (XII) a polynudeotide that encodes a 109P1 D4-related protein that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100% identical to an entire amino acid sequence shown in Figure 2A-I; (XIII) a polynucleotide that encodes at least one peptide set forth in Tables VIII-XXI and XXII-XLIX; (XIV) a polynudeotide that encodes a peptide region of at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acids of a peptide of Figures 3A in any whole number increment up to 1021 that Includes at least 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14,15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having a value greater than in the Hydrophilicity profile of Figure (XV) a polynucleotide that encodes a peptide region of at least 5, 6, 7, 8, 9, 10, 11, 12,13,14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30,31, 32, 33, 34, 35 amino acds of a peptide of Figure 3A in any whole number increment up to 1021 that indudes 1, 2, 3,4, 5, 6, 7, 8, 9,10,11, 12, 13,14, 15, 16,17, 18,19, 20,21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid positiori(s) having a value less than 0.5 in the Hydropathlcity profile of Figure 6; (XVI) a polynucleotide that encodes a peptide region of at least 5, 6, 7, 8, 9,10, 11, 12,13, 14,15, 16,17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino adds of a peptide of Figure 3A in any whole number increment up to 1021 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino add position(s) having a value greater than 0.5 in the Percent Accessible Residues profile of Figure 7; (XVII) a polynucleotide that encodes a peptide region of at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino adds of a peptide of Figure 3A in any whole number increment up to 1021 that includes 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acd position(s) having a value greater than 0.5 in the Average Flexibility profile of Figure 8; (XVIII) a polynucleotide that encodes a peptide region of at least 5, 6, 7, 8, 9, 10, 11, 12, 13,14,15,16, 17, r 22 00 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acids of a peptide of Figure 3A in any whole number increment up to 1021 that includes 1, 2,3, 4, 5, 6, 7, 8, 9,10,11,12,13,14, 15,16, 17, 18,19, rI 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having a value greater than 0.5 In the Beta-turn profile of Figure 9; (XIX) a polynudeotide that encodes a peptide region of at least 5, 6, 7, 8, 9, 10,11, 12, 13,14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acids of a peptide of Figure 3B, 3C, and/or 3D in any whole number increment up to 1054, 1347, and/or 1337 respectively that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having a value greater than 0.5 in the Hydrophilicity profile of Figure (XX) a polynucleoide that encodes a peptide region of at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino adds of a peptide of Figure 3B, 3C, and/or 030 3D In any whole number Increment up to 1054,1347, and/or 1337 respectively tha Includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 13, 14, 15,16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having a value less than 0.5 In the Hydropathlcity profile of Figure 6; (XXI) a polynudeotide that encodes a pepbde region of at least 5, 6,7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 31, 32, 33, 34, 35 amino acids of a peptide of Figure 3B, 3C, and or 3D in any whole number Increment up to 1054, 1347, and/or 1337 respectively that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12,13, 14,15,16, 17,18,19, 20, 21, 22, 23, 24,25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino add position(s) having a value greater than 0.5 in the Percent Accessible Residues profile of Figure 7; (XXII) a polynudeoide that encodes a peptide region of at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acids of a peptide of Figure 3B, 3C, and/or 3D in any whole number Increment up to 1054, 1347, and/or 1337 respectively that includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having a value greater than 0.5 In the Average Fledbility profile of Figure 8; (XXIII) a polynudeotide that encodes a peptide region of at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acds of a pepide of Figure 3B, 3C, and/or 3D in any whole number increment up to 1054, 1347, and/or 1337 respectively that Includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12,13,14, 15,16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having a value greater than 0.5 in the Beta-turn profile of Figure 9; (XXIV) a polynucleotide that encodes a peptide region of at least 5,6, 7, 8, 9, 10, 11, 12, 13,14,15, 16,17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32 33, 34, 35 amino acids of a peptide of Figure 3E, 3F, 3G, 3H and/or 31 in any whole number increment up to 1310, 1037, 1048, 1340, and/or 1037 respectively th at includes 1, 2, 3,4, 5, 6, 7, 8,9,10,11,12,13,14,15,16,17,18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33,34, amino acid positon(s) having a value greater than 0.6 in the Hydrophilicity profile of Figure (XXV) a polynucleotide that encodes a peptide region of at least 5, 6, 7, 8, 9, 10,11, 12,13,14,15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acids of a peptide of Figure 3E, 3F, 3G, 3H and/or 31 In any whole number Increment up to 1310, 1037, 1048, 1340, and/or 1037 respectively that Indudes 1, 2,3, 4, 5,6, 7,8,9, 10, 11, 12,13, 14, 15,16, 17,18,19,20, 21, 22,23, 24,25, 26,27,28, 29, 30, 31,32, 33,34, amino acid position(s) having a value less than 0.5 In the Hydropathicity profile of Figure 6; 23 00 (XXVI) a polynucleotide that encodes a peptide region of at least 5, 6, 7, 8, 9,10, 11, 12,13,14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30,31, 32, 33, 34, 35 amino acids of a peptide of Figure 3E, 3F, 3G, 3H and/or 31 in any whole number increment up to 1310, 1037, 1048, 1340, and/or 1037 respectively that includes 1, d) 2, 3, 4, 5,6, 7, 8, 9, 10, 11,12,13,14,15,16, 17, 18, 19, 20, 21, 22, 23, 24,25, 26, 27, 28, 29, 30, 31, 32, 33, 34, amino acid position(s) having a value greater than 0.5 in the Percent Accessible Residues profile of Figure 7; (XXVII) a polynucleotide that encodes a peptide region of at least 5, 6, 7, 8, 9,10,11,12,13,14,15,16, 17,18, 19, 20,21,22, 23, 24,25, 26, 27, 28,29, 30,31,32, 33,34, 35 amino acids of a peptide of Figure 3E, 3F, 3G, 3H and/or 31 in any whole number increment up to 1310, 1037, 1048, 1340, and/or 1037 respectively that indudes 1, CN 2, 3,4, 5, 6, 7, 8, 9,10, 11, 12,13,14,15,16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, C' 35 amino add position(s) having a value greater than 0.5 in the Average Flexibility profile of Figure 8; C (XXVIII) a polynudeotide that encodes a peptide region of at least 5, 6, 7, 8, 9, 10, 11, 12 13,14,15,16, 17, 18, 00 S19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino adds of a peptide of Figure 3E, 3F, 3G, 3H, 0and/or 31 In any whole number increment up to 1310, 1037,1048, 1340, and/or 1037 respectively that includes 1, 2, 3,4, 5, 6,7, 8, 9,10,11,12,13,14,15,16,17,18, 19,20, 21, 22, 23, 24,25, 26,27,28, 29, 30, 31, 32,33, 34, amino add position(s) having a value greater than 0.5 in the Beta-turn profile of Figure 9; (XXIX) a polynudeotide that is fully complementary to a polynudeotide of any one of (I)-(XXVIII); (XXX) a polynudeotide that is fully complementary to a polynudeotide of any one of (I)-(XXIX); (XXXI) a peptide that is encoded by any of to (XXX); and; (XXXII) a composition comprising a polynudeotide of any of (IHXXX) or peptide of (XXXI) together with a pharmaceutical excipient and/or in a human unit dose form; (XXXIII) a method of using a polynudeotide of any or peptide of (XXXI) or a composition of (XXXII) in a method to modulate a cell expressing 109P1D4; (XXXIV) a method of using a polynudeotide of any or peptide of (XXXI) or a composition of (XXXII) in a method to diagnose, prophylax, prognose, or treat an individual who bears a cell expressing 109P1D4; (XXXV) a method of using a polynudeotide of any (I)-XXX) or peptide of (XXXI) or a composition of (XXXll) in a method to diagnose, prophylax, prognose, or treat an individual who bears a cell expressing 109P1D4, said cell from a cancer of a tissue listed in Table I; (XXXVI) a method of using a polynudeotide of any or peptide of (XXXI) or a composition of (XXXII) in a method to diagnose, prophylax, prognose, or treat a a cancer; (XXXVII) a method of using a polynudeotide of any or peptide of (XXXI) or a composition of (XXXII) in a method to diagnose, prophylax, prognose, or treat a a cancer of a tissue listed in Table I; and; (XXXVIII) a method of using a polynudeotide of any (I)-XXX) or peptide of (XXXI) or a composition of (XXXII) In a method to identify or characterize a modulator of a cell expressing 109P1D4.

As used herein, a range is understood to disdose specifically all whole unit positions thereof.

Typical embodiments of the Invention disclosed herein include 109P1D4 polynuceotides that encode specific portions of 109P1D4 mRNA sequences (and those which are complementary to such sequences) such as those that encode the proteins and/or fragments thereof, for example: O0 4, 5, 6, 7, 8, 9, 10,11, 12, 13,14,15,16,17,18,19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45,50, 55, 60, 65, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 725, 750, 775, 800, 825, 850, 875, 900, 925, 950, 975, 1000, 1010, 1020, and 1021 or more contiguous amino acids of 109P1D4 variant 1; the Smaximal lengths relevant for other variants are: variant 2, 1054 amino adds; variant 3, 1347 amino acids, variant 4, 1337 Samino acids, variant 5, 1310 amino acids, variant 6; 1047 amino adds, variant 7; 1048 amino acids, variant 8; 1340 amino Sacids and variant 9; 1037 amoni adds.

For example, representative embodiments of the invention disdosed herein include: polynudeotides and their O encoded peptides themselves encoding about amino add 1 to about amino acid 10 of the 109P1D4 protein shown in Figure 2 or Figure 3, polynucleotides encoding about amino acid 10 to about amino acid 20 of the 109P1D4 protein shown in Figure C 2 or Figure 3, polynucleolides encoding about amino add 20 to about amino acid 30 of the 109P1D4 protein shown in Figure NC= 2 or Figure 3, polynucleotides encoding about amino add 30 to about amino add 40 of the 109P1D4 protein shown in Figure 00 2 or Figure 3, polynucleoides encoding abou amino acid 40 to about amino add 50 of the 109P1D4 protein shown in Figure 2 or Figure 3, polynucleotides encoding about amino acid 50 to about amino acid 60 of the 109P1 D4 protein shown in Figure 2 or Figure 3, polynucleotides encoding about amino add 60 to about amino acid 70 of the 109P1 D4 protein shown in Figure 2 or Figure 3, polynucleotides encoding about amino acid 70 to about amino add 80 of the 109P1 D4 protein shown in Figure 2 or Figure 3, polynudeotides encoding about amino acid 80 to about amino acid 90 of the 109P1 D4 protein shown in Figure 2 or Figure 3, polynudeotides encoding about amino acid 90 to about amino acid 100 of the 109P1 D4 protein shown in Figure 2 or Figure 3, in increments of about 10 amino adds, ending at the carboxyl terminal amino acid set forth in Figure 2 or Figure 3. Accordingly, polynucleotides encoding portions of the amino add sequence (of about 10 amino adds), of amino adds, 100 through the carboxyl terminal amino acid of the 109P1 D4 protein are embodiments of the invention. Wherein it is understood that each particular amino acid position discloses that position plus or minus five amino acid residues.

Polynucleotides encoding relatively long portions of a 109P1D4 protein are also within the scope of the invention.

For example, polynucleotides encoding from about amino acid 1 (or 20 or 30 or 40 etc.) to about amino acid 20, (or 30, or or 50 etc.) of the 109P1D4 protein "or variant" shown in Figure 2 or Figure 3 can be generated by a variety of techniques well known in the art. These polynucleotide fragments can include any portion of the 109P1D4 sequence as shown in Figure 2.

Additional illustrative embodiments of the invention disclosed herein include 109P1D4 polynucleotide fragments encoding one or more of the biological motifs contained within a 109P1D4 protein 'or variant" sequence, including one or more of the motif-bearing subsequences of a 109P1D4 protein "or variant set forth in Tables VIII-XX and XXII-XLIX. In another embodiment, typical polynudeotide fragments of the invention encode one or more of the regions of 109P1D4 protein or variant that exhibit homology to a known molecule. In another embodiment of the invention, typical polynucleotide fragments can encode one or more of the 109P1D4 protein or variant N-glycosylation sites, cAMP and cGMP-dependent protein kinase phosphorylation sites, casein kinase II phosphorylation sites or N-myristoylation site and amidation sites.

Note that to determine the starting position of any peptide set forth in Tables VIII-XXI and Tables XXII to XLIX (collectively HLA Peptide Tables) respective to its parental protein, variant 1, variant 2, etc., reference is made to three factors: the particular variant, the length of the peptide in an HLA Peptide Table, and the Search Peptides listed in Table VII.

Generally, a unique Search Peptide is used to obtain HLA peptides for a particular variant The position of each Search Peptide relative to its respective parent molecule is listed in Table VII. Accordingly, if a Search Peptide begins at position one must add the value "X minus 1" to each position In Tables VIII-XXI and Tables XXII-IL to obtain the actual position of the HLA peptides in their parental molecule. For example if a particular Search Peptide begins at position 150 of its parental molecule, one must add 150 1, 149 to each HLA peptide amino add position to calculate the position of that amino add 00 in the parent molecule.

IIA.) Uses of 109P1D4 Polynucleotides Cr IIA1.) Monitoring of Genetic Abnormalities The polynuceotides of the preceding paragraphs have a number of different specific uses. The human 109P1D4 gene maps to the chromosomal location set forth in the Example entitled "Chromosomal Mapping of 109P1 D4." For t~ example, because the 109P1D4 gene maps to this chromosome, polynudeotides that encode different regions of the 109P1D4 proteins are used to characterize cytogenetic abnormalities of this chromosomal locale, such as abnormalities that are identified as being associated with various cancers. In certain genes, a variety of chromosomal abnormalities including Srearrangements have been identified as frequent cytogenetic abnormalities in a number of different cancers (see e.g.

0 Krajinovic et al., Mutat Res. 382(3-4): 81-83 (1998); Johansson et al., Blood 86(10): 3905-3914 (1995) and Finger et al., P.NAS. 85(23): 9158-9162 (1988)). Thus, polynudeotides encoding specific regions of the 109P1 D4 proteins provide new CN tools that can be used to delineate, with greater precision than previously possible, cytogenetic abnormalities in the 00 chromosomal region that encodes 109P1D4 that may contribute to the malignant phenotype. In this context, these Spolynudeotides satisfy a need in the art for expanding the sensitivity of chromosomal screening in order to identify more subtle and less common chromosomal abnormalities (see e.g. Evans et al., Am. J. Obstet Gynecol 171(4): 1055-1057 (1994)).

Furthermore, as 109P1 D4 was shown to be highly expressed in prostate and other cancers, 109P1D4 polynucleotides are used in methods assessing the status of 109P1D4 gene products in normal versus cancerous tissues.

Typically, polynucleotides that encode specific regions of the 109P1D4 proteins are used to assess the presence of perturbations (such as deletions, insertions, point mutations, or alterations resulting in a loss of an antigen etc.) in specific regions of the 109P1D4 gene, such as regions containing one or more motifs. Exemplary assays Include both RT-PCR assays as well as single-strand conformation polymorphism (SSCP) analysis (see, Marrogi at al., J. Cutan. Pathol.

26(8): 369-378 (1999), both of which utilize polynudeotides encoding specific regions of a protein to examine these regions within the protein.

IIA.2.) Antisense Embodiments Other specifically contemplated nucleic add related embodiments of the invention disclosed herein are genomic DNA, cDNAs, nbozymes, and antisense molecules, as well as nucleic add molecules based on an alternative backbone; or induding alterative bases, whether derived from natural sources or synthesized, and include molecules capable of inhibiting the RNA or protein expression of 109P1 D4. For example, antisense molecules can be RNAs or other molecules, induding peptide nudeic acids (PNAs) or non-nucleic add molecules such as phosphorothloate derivatives that specifically bind DNA or RNA in a base pair-dependent manner. A skilled artisan can readily obtain these classes of nucleic add molecules using the 109P1D4 polynudeotides and polynuceotide sequences disclosed herein.

Antisense technology entails the administration of exogenous oligonucleotides that bind to a target polynucleotide located within the cells. The term "antisense' refers to the fact that such oligonudeotides are complementary to their intracellular targets, 109P1D4. See for example, Jack Cohen, Oligodeoxynucleotides, Antisense Inhibitors of Gene Expression, CRC Press, 1989; and Synthesis 1:1-5 (1988). The 109P1D4 antisense oligonucleotides of the present invention include derivatives such as S-oligonudeotides (phosphorothioate derivatives or S-ollgos, see, Jack Cohen, supra), which exhibit enhanced cancer cell growth inhibitory action. S-oligos (nudeoside phosphorothloates) are isoelectronic analogs of an oligonucleotide (0-oligo) in which a nonbridging oxygen atom of the phosphate group is replaced by a sulfur atom. The S-oligos of the present invention can be prepared by treatment of the corresponding O-oligos with 3H-1,2benzodithiol-3-one-1,1-dioxide, which is a sulfur transfer reagent See, lyer, R. P. et at, J. Org. Chem. 55:4693-4698 OO (1990); and lyer, R. P. et J. Am. Chem. Soc. 112:1253-1254 (1990). Additional 109P1D4 antisense oligonucleotides of O the present invention include morpholino antisense oligonucleotides known in the art (see, Partridge et al., 1996, C Antisense Nucleic Acid Drug Development 6:169-175).

O The 109P1D4 antisense digonudeotides of the present invention typically can be RNA or DNA that is complementary to and stably hybridizes with the first 100 5' codons or last 100 3' codons of a 109P1 D4 genomic sequence V or the corresponding mRNA. Absolute complementarity Is not required, although high degrees of complementarity are preferred. Use of an oligonucleotide complementary to this region allows for the selective hybridization to 109P1D4 mRNA and not to mRNA specifying other regulatory subunits of protein kinase. In one embodiment, 109P1D4 antisense oligonudeotides of the present Invention are 15 to 30-mer fragments of the antisense DNA molecule that have a sequence that hybridizes to 109P1 D4 mRNA. Optionally, 109P1 D4 antisense oligonucleotide is a 30-mer oligonucleotide that is complementary to a region in the first 10 5' codons or last 10 3' codons of 109P1D4. Alternatively, the antisense molecules C are modified to employ ribozymes in the inhibition of 109P1D4 expression, see, L A. Couture D. T. Stinchcomb; STrends Genet 12: 510-515 (1996).

SII.A.3.) Primers and Primer Pairs Further specific embodiments of these nucleotides of the invention include primers and primer pairs, which allow the specific amplification of polynudeotides of the invention or of any specific parts thereof, and probes that selectively or specifically hybridize to nucleic acid molecules of the invention or to any part thereof. Probes can be labeled with a detectable marker, such as, for example, a radioisotope, fluorescent compound, bioluminescent compound, a chemiluminescent compound, metal chelator or enzyme. Such probes and primers are used to detect the presence of a 109P1D4 polynuceotide in a sample and as a means for detecting a cell expressing a 109P1D4 protein.

Examples of such probes include polypeptides comprising all or part of the human 109P1 D4 cDNA sequence shown in Figure 2 Examples of primer pairs capable of specifically amplifying 109P1 4 mRNAs are also described in the Examples. As will be understood by the skilled artisan, a great many different primers and probes can be prepared based on the sequences provided herein and used effectively to amplify and/or detect a 109P1D4 mRNA.

The 109P1 D4 polynucleotides of the invention are useful for a variety of purposes, incuding but not limited to their use as probes and primers for the amplification and/or detection of the 109P1D4 gene(s), mRNA(s), or fragments thereof; as reagents for the diagnosis and/or prognosis of prostate cancer and other cancers; as coding sequences capable of directing the expression of 109P1 D4 polypeptides; as tools for modulating or inhibiting the expression of the 109P1 D4 gene(s) and/or translation of the 109P1 D4 transcript(s); and as therapeutic agents.

The present invention includes the use of any probe as described herein to identify and isolate a 109P1D4 or 109P1D4 related nucleic acid sequence from a naturally occurring source, such as humans or other mammals, as well as the isolated nucleic add sequence perse, which would comprise all or most of the sequences found in the probe used.

II.A.4.) Isolation of 109P1D4-Encodlng Nucleic Add Molecules The 109P1D4 cDNA sequences described herein enable the isolation of other polynucleotides encoding 109P1D4 gene product(s), as well as the isolation of polynudeotides encoding 109P1D4 gene product homologs, altematively spliced isoforms, allelic variants, and mutant forms of a 109P1D4 gene product as well as polynucleolides that encode analogs of 109P1 D4-related proteins. Various molecular cloning methods that can be employed to isolate full length cDNAs encoding a 109P1D4 gene are well known (see, for example, Sambrook, J. et Molecular Cloning: A Laboratory Manual, 2d edition, Cold Spring Harbor Press, New York, 1989; Current Protocols in Molecular Biology. Ausubel et al., Eds., Wiley and Sons, 1995). For example, lambda phage cloning methodologies can be conveniently employed, using commercially available doning systems Lambda ZAP Express, Stratagene). Phage clones containing 109P1D4 gene cDNAs can be identified by probing with a labeled 109P1D4 00 cDNA or a fragment thereof. For example, in one embodiment, a 109P1D4 cDNA Figure 2) or a portion thereof can be Ssynthesized and used as a probe to retrieve overlapping and full-length cDNAs corresponding to a 109P1D4 gene. A 109P1D4 Sgene itself can be isolated by screening genomic DNA libraries, bacterial artificial chromosome libraries (BACs), yeast artificial chromosome libraries (YACs), and the like, with 109P1 D4 DNA probes or primers.

Recombinant Nucleic Acd Molecules and Host-Vector Systems in The invention also provides recombinant DNA or RNA molecules containing a 109P1D4 polynucleotide, a fragment analog or homologue thereof, including but not limited to phages, plasmlds, phagemids, cosmids, YACs, BACs, as well as various viral and non-viral vectors well known in the art, and cells transformed or transfected with such recombinant DNA or RNA Smolecules. Methods for generating such molecules are well known (see, for example, Sambrook et 1989, supra).

SThe invention further provides a host-vector system comprising a recombinant DNA molecule containing a 109P1D4 polynudeotide, fragment, analog or homologue thereof within a suitable prokaryotic or eukaryotic host cell. Examples of CN suitable eukaryotic host cells include a yeast cell, a plant cell, or an animal cell, such as a mammalian cell or an insect cell 00 a baculovlrus-infectible cell such as an Sf9 or HighFive cell). Examples of suitable mammalian cells Include various Sprostate cancer cell lines such as DU145 and TsuPrl, other transfectable or transduclble prostate cancer cell lines, primary cells (PrEC), as well as a number of mammalian cells routinely used for the expression of recombinant proteins COS, CHO, 293, 293T cells). More particularly, a polynudeotide comprising the coding sequence of 109P1D4 or a fragment, analog or homolog thereof can be used to generate 109P1D4 proteins or fragments thereof using any number of host-vector systems routinely used and widely known in the art A wide range of host-vector systems suitable for the expression of 109P1D4 proteins or fragments thereof are available, see for example, Sambrook et 1989, supra; Current Protocols in Molecular Biology, 1995, supra). Preferred vectors for mammalian expression include but are not limited to pcDNA 3.1 myc-His-tag (Invitrogen) and the retroviral vector pSRatkneo (Muller et al., 1991, MCB 11:1785). Using these expression vectors, 109P1D4 can be expressed in several prostate cancer and non-prostate cell lines, including for example 293, 293T, rat-1, NIH 3T3 and TsuPrl. The host-vector systems of the invention are useful for the production of a 109P1 D4 protein or fragment thereof. Such host-vector systems can be employed to study the functional properties of 109P1D4 and 109P1D4 mutations or analogs.

Recombinant human 109P1 D4 protein or an analog or homolog or fragment thereof can be produced by mammalian cells transfected with a construct encoding a 109P1D4-related nudeotide. For example, 293T cells can be transfected with an expression plasmid encoding 109P1D4 or fragment analog or homolog thereof, a 109P1D4-related protein is expressed in the 293T cells, and the recombinant 109P1D4 protein is isolated using standard purification methods affinity purification using anti-109P1D4 antibodies). In another embodiment a 109P1D4 coding sequence is subdoned into the retroviral vector pSRaMSVtkneo and used to infect various mammalian cell lines, such as NIH 3T3, TsuPrl, 293 and rat-1 in order to establish 109P1 D4 expressing cell lines. Various other expression systems well known in the art can also be employed. Expression constructs encoding a leader peptide joined in frame to a 109P1D4 coding sequence can be used for the generation of a secreted form of recombinant 109P1 D4 protein.

As discussed herein, redundancy in the genetic code permits variation In 109P1D4 gene sequences. In particular, it is known in the art that specific host species often have specific codon preferences, and thus one can adapt the disclosed sequence as preferred for a desired host For example, preferred analog codon sequences typically have rare codons codons having a usage frequency of less than about 20% in known sequences of the desired host) replaced with higher frequency codons. Codon preferences for a specific species are calculated, for example, by utilizing codon usage tables available on the INTERNET such as at URL dnaaffrc.go.jp/-nakamura/codon.html.

Additional sequence modifications are known to enhance protein expression in a cellular host These include 28 00 elimination of sequences encoding spurious polyadenylation signals, exonlintron splice site signals, transposon-like repeats, and/or other such well-characterized sequences that are deleterious to gene expression. The GC content of the sequence is r1 adjusted to levels average for a given cellular host, as calculated by reference to known genes expressed in the host cell.

Where possible, the sequence is modified to avoid predicted hairpin secondary mRNA structures. Other useful modifications include the addition of a translational initiation consensus sequence at the start of the open reading frame, as described in f Kozak, Mol. Cell Biol., 9:5073-5080 (1989). Skilled artisans understand that the general rule that eukaryotic ribosomes initiate translation exclusively at the 5' proximal AUG codon is abrogated only under rare conditions (see, Kozak PNAS 92(7): 2662-2666, (1995) and Kozak NAR 15(20): 8125-8148 (1987)).

IIl.) 109P1D4-related Proteins Another aspect of the present invention provides 109P1D4-related proteins. Specific embodiments of 109P1D4 6 proteins comprise a polypeptide having all or part of the amino add sequence of human 109P1D4 as shown in Figure 2 or 00 0 Figure 3. Alternatively, embodiments of 109P1 D4 proteins comprise variant homolog or analog polypeptides that have Salterations in the amino add sequence of 109P1 D4 shown in Figure 2 or Figure 3.

Embodiments of a 109P1D4 polypeptide indude: a 109P1D4 polypeptide having a sequence shown in Figure 2, a peptide sequence of a 109P1 D4 as shown in Figure 2 wherein T is U; at least 10 contiguous nucleotides of a polypeptide having the sequence as shown in Figure 2; or, at least 10 contiguous peptides of a polypeptide having the sequence as shown in Figure 2 where T Is U. For example, embodiments of 109P1D4 peptides comprise, without limitation: a protein comprising, consisting essentially of, or consisting of an amino acid sequence as shown in Figure 2A-I or Figure 3A-I; (II) a 109P1D4-related protein that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100% homologous to an entire amino add sequence shown in Figure 2A-I or 3A-I; (III) a 109P1D4-related protein that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100% identical to an entire amino acid sequence shown in Figure 2A-I or 3A-I; (IV) a protein that comprises at least one peptide set forth In Tables VIII to XLIX, optionally with a proviso that it is not an entire protein of Figure 2; a protein that comprises at least one peptide set forth in Tables VIII-XXI, collectively, which peptide is also set forth in Tables XXII to XLIX, collectively, optionally with a proviso that it is not an entire protein of Figure 2; (VI) a protein that comprises at least two peptides selected from the peptides set forth in Tables VIII-XLIX, optionally with a proviso that it is not an entire protein of Figure 2; (VII) a protein that comprises at least two peptides selected from the peptides set forth in Tables VIII to XLIX collectively, with a proviso that the protein is not a contiguous sequence from an amino add sequence of Figure 2; (VIII) a protein that comprises at least one peptide selected from the peptides set forth in Tables VIII-XXJ; and at least one peptide selected from the peptides set forth in Tables XXII to XLIX, with a proviso that the protein is not a contiguous sequence from an amino acid sequence of Figure 2; (IX) a polypeptide comprising at least 5, 6, 7, 8, 9, 10,11, 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 26, 27, 28, 29, 30,31, 32, 33, 34, 35 amino adds of a protein of Figure 3A, 3B, 3C, 3D and/or 3E in any whole number increment up to 1021, 1054, 1347, 1337, and/or 1310 respectively that includes at least 1, 2, 3,4, 00 6, 7, 8, 9,10,11, 12,13,14, 15,16,17,18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having a value greater than 0.5 in the Hydrophilicity profile of Figure a polypeptide comprising at least 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acids of a protein of Figure 3A, 3B, 3C, 3D, and/or 3E, In any whole number increment up to 1021, 1054, 1347, 1337, and/or 1310 respectively respectively that includes at least V at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having a value less than 0.5 in the Hydropathicity profile of Figure 6; S(XI) a polypeptide comprising at least 5, 6, 7, 8, 9,10,11, 12,13,14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 26, 27, 28,29, 30, 31, 32, 33, 34, 35 amino acids of a protein of Figure 3A, 3B, 3C, 3D, and/or 3E, in any whole r number increment up to 1021, 1054, 1347, 1337, and/or 1310 respectively respectively that indudes at least at rK least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15,16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 0 33, 34, 35 amino acid position(s) having a value greater than 0.5 in the Percent Accessible Residues profile of SFigure 7; (XII) a polypeptide comprising at least 5, 6, 7, 8, 9,10,11, 12,13,14,15,16,17, 18, 19,20, 21, 22, 23, 24, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acids of a protein of Figure 3A, 3B, 3C, 3D, and/or 3E, in any whole number increment up to 1021, 1054, 1347, 1337, and/or 1310 respectively respectively that indudes at least at least 1, 2, 3,4, 5, 6, 7, 8, 9, 10,11, 12,13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having a value greater than 0.5 in the Average Flexibility profile of Figure 8; (XIII) a polypeptide comprising at least 5, 6, 7, 8, 9,10, 11, 12,13, 14,15,16, 17, 18, 19, 20, 21, 22, 23, 24, 26, 27, 28, 29, 30, 31, 32, 33, 34, amino adds of a protein of Figure 3A, 3B, 30, 3D, and 3E in any whole number increment up to 1021, 1054, 1347, 1337, and/or 1310 respectively respectively that indudes at least at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acd position(s) having a value greater than 0.5 in the Beta-tur profile of Figure 9; (XIV) a polypeptide comprising at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino adds of a protein of Figure 3F, 3G, 3H, and/or 31, in any whole number increment up to 1037, 1048, 1340, and/or 1037 respectively that includes at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12,13,14, 15,16,17,18,19,20,21, 22, 23, 24, 25,26,27,28,29,30,31, 32,33,34,35 amino acid position(s) having a value greater than 0.5 in the Hydrophilicty profile of Figure (XV) a polypeptide comprising at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino adds of a protein of Figure 3F, 3G, 3H, and/or 31 In any whole number Increment up to 1037, 1048, 1340, and/or 1037 respectively that includes at least at least 1, 2, 3, 4, 5, 6,7, 8,9, 10,11, 12,13,14,15,16,17,18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having a value less than 0.5 in the Hydropathicity profile of Figure 6; (XVI) a polypeptide comprising at least 5, 6, 7, 8, 9, 10, 11, 12,13, 14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acids of a protein of Figure 3F, 3G, 3H, and/or 31 in any whole number Increment up to 1037, 1048, 1340, and/or 1037 respectively that indudes at least at least 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino add position(s) having a value greater than 0.5 In the Percent Accessible Residues profile of Figure 7; (XVII) a polypeptide comprising at least 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 00 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 amino acids of a protein of Figure 3F, 3G, 3H, and/or 31 in any whole number increment up to 1037, 1048, 1340, and/or 1037 respectively that includes at least at least 1, 2, 3, 4, 5, 6, 7, C( 8, 9, 10, 11, 12 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23,24,25,26, 27, 28. 29, 30, 31, 32, 33, 34, 35 amino acid position(s) having a value greater than 0.5 in the Average Flexibility profile of Figure 8; (XVIII) a polypeptide comprising at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 26, 27, 28, 29, 30, 31, 32, 33, 34, amino acids of a protein of Figure 3F, 3G, 3H, and/or 31 In any whole number increment up to 1037, 1048, 1340, and/or 1037 respectively that includes at least at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,25,26, 27, 28, 29, 30, 31,32, 33, 34, 35 amino acid C- position(s) having a value greater than 0.5 in the Beta-turn profile of Figure 9; K (XIX) a peptide that occurs at least twice in Tables VIIIXXI and XX)I to XLIX, collectively; 0 (XX) a peptide that occurs at least three times In Tables VIII-XXI and XXII to XLIX, collectively; (XX) a peptide that occurs at least four times in Tables VIII-XXI and XXII to XLIX, collectively (XXI) a peptide that occurs at least five times in Tables VIll-XXI and XXII to XLIX, collectively; (XXII) a peptide that occurs at least once times in Tables VIII-XXI and at least once in tables XXII to collectivelyIX; (XXIV) a peptide that occurs at least once in Tables VIII-XXI, and at least twice in tables XXII to XLIX; (XXV) a peptide that occurs at least once in Tables VIII-XXI, and at least once in tables XXII to XLIX; (XXV) a peptide that occurs at least twice in Tables VIII-XXI, and at least twice in tables XXII to XLIX; (XXV) a peptide that occurs at least twice in Tables VIII-XXI, and at least twice in tables XXII to XLIX; (XXVII) a peptide which comprises one two, three, four, or five of the following characteristics, or an oligonudeotide encoding such peptide: i) a region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to the full length of that protein in Figure 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Hydrophilicity profile of Figure ii) a region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to the full length of that protein in Figure 3, that includes an amino add position having a value equal to or less than 0.5, 0.4, 0.3, 0.2, 0.1, or having a value equal to 0.0, in the Hydropathicity profile of Figure 6; iii) a region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to the full length of that protein in Figure 3, that includes an amino add position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, In the Percent Accessible Residues profile of Figure 7; iv) a region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to the full length of that protein in Figure 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Average Flexibility profile of Figure 8; or, v) a region of at least 5 amino acids of a particular peptide of Figure 3, In any whole number increment up to the full length of that protein in Figure 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Beta-tur profile of Figure 9;; (XXVIII) a composition comprising a peptide of ()-(XXVII) or an antibody or binding region thereof together with a pharmaceutical exciplent and/or in a human unit dose form.

00 (XXIX) a method of using a peptide of (IHXXVII), or an antibody or binding region thereof or a composition of (XXVIII) in a method to modulate a cell expressing 109P1D4,; (XXX) a method of using a peplide of (IHXXVII) or an antibody or binding region thereof or a composition of

S(XXVI

l I) in a method to diagnose, prophylax, prognose, or treat an individual who bears a cell expressing 109P1D4; (XXXI) a method of using a peptide of (I)-(XXVII) or an antibody or binding region thereof or a composition (XXVIII) in a method to diagnose, prophylax, prognose, or treat an individual who bears a cell expressing 0 109P1D4, said cell from a cancer of a tissue listed in Table I; 0 (XXXII) a method of using a peptide of (I)-(XXVll) or an antibody or binding region thereof or a composition of (XXVIII) in a method to diagnose, prophylax, prognose, or treat a a cancer; 00 (XXXIII) a method of using a peptide of (I)-(XXVII) or an antibody or binding region thereof or a composition of (XXVIII) in a method to diagnose, prophylax, prognose, or treat a a cancer of a tissue listed in Table I; and; (XXXIV) a method of using a a peptide of (I)-(XXVll) or an antibody or binding region thereof or a composition (XXVIII) in a method to identify or characterize a modulator of a cell expressing 109P1 04 As used herein, a range is understood to specifically disclose all whole unit positions thereof.

Typical embodiments of the invention disclosed herein Include 109P1D4 polynucleotides that encode specific portions of 109P1D4 mRNA sequences (and those which are complementary to such sequences) such as those that encode the proteins and/or fragments thereof, for example: 4, 5, 6, 7, 8, 9,10. 11, 12, 13, 14,15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45,50, 55, 60, 65, 80,85, 90,95,100, 105,110,115,120,125,130,135,140,145,150, 155, 160,165, 170, 175, 180,185,190, 195, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 725, 750, 775, 800, 825, 850, 875, 900, 925, 950, 975, 1000, 1010, 1020, and 1021 or more contiguous amino acids of 109P1D4 variant 1; the maximal lengths relevant for other variants are: variant 2, 1054 amino acids; variant 3, 1347 amino adds, variant 4, 1337 amino adds, variant 5, 1310 amino acids, variant 6; 1037 amino adds, variant 7; 1048 amino adds, variant 8; 1340 amino adds, and variant 9; 1037 amino acids..

.In general, naturally occurrning allelic variants of human 109P1 D4 share a high degree of structural identity and homology 90% or more homology). Typically, allelic variants of a 109P1 D4 protein contain conservative amino add substitutions within the 109P1D4 sequences described herein or contain a substitution of an amino add from a corresponding position in a homologue of 109P1D4. One class of 109P1D4 allelic variants are proteins that share a high degree of homology with at least a small region of a particular 109P1D4 amino acid sequence, but further contain a radical departure from the sequence, such as a non-conservative substitution, truncation, insertion or frame shift. In comparisons of protein sequences, the terms, similarity, identity, and homology each have a distinct meaning as appreciated in the field of genetics. Moreover, orthology and paralogy can be important concepts describing the relationship of members of a given protein family In one organism to the members of the same family In other organisms.

Amino acid abbreviations are provided in Table II. Conservative amino add substitutions can frequently be made in a protein without altering either the conformation or the function of the protein. Proteins of the Invention can comprise 1, 2, 3,4, 5, 6, 7, 8, 9,10,11,12,13,14,15 conservative substitutions. Such changes include substituting any of isoleudne valine and leudne for any other of these hydrophobic amino acids; aspartic add for glutamic add and vice versa; glutamine for asparagine and vice versa; and serine for threonine and vice versa. Other substitutions

I

00 can also be considered conservative, depending on the environment of the particular amino acid and its role in the threedimensional structure of the protein. For example, glydne and alanine can frequently be Interchangeable, as can S alanine and valine Methionine which Is relatively hydrophobic, can frequently be interchanged with leucine and isoleucne, and sometimes with valine. Lysine and arginine are frequently interchangeable in locations in which the C/significant feature of the amino acid residue is its charge and the differing pK's of these two amino acid residues are not V significant Still other changes can be considered 'conservative' in particular environments (see, e.g. Table III herein; pages 13-15 "Biochemistry" 2nd ED. Lubert Stryer ed (Stanford University); Henikoff et PNAS 1992 Vol 89 10915-10919; Lei et at., J Biol Chem 1995 May 19; 270(20):11882-6).

SEmbodiments of the invention disclosed herein include a wide variety of art-accepted variants or analogs of S109P1D4 proteins such as polypeptides having amino add insertions, deletions and substitutions. 109P104 variants can be made using methods known in the art such as site-directed mutagenesis, alanine scanning, and PCR mutagenesis. Sitedirected mutagenesis (Carter et al., Nuc. Acids Res., 13:4331 (1986); Zoller el al., Nucl. Acids Res., 10.6487 (1987)), 00 Scassette mutagenesis (Wells et al., Gene, 34:315 (1985)), restriction selection mutagenesis (Wells et al., Philos. Trans. R.

SSoc. London SerA, 317:415 (1986)) or other known techniques can be performed on the doned DNA to produce the 109P1D4 variant DNA.

Scanning amino add analysis can also be employed to identify one or more amino acids along a contiguous sequence that is involved in a specific biological activity such as a protein-protein interaction. Among the preferred scanning amino acids are relatively small, neutral amino adds. Such amino adds include alanine, glycine, serine, and cysteine.

Alanine is typically a preferred scanning amino acid among this group because it eliminates the side-chain beyond the betacarbon and is less likely to alter the main-chain conformation of the variant Alanine is also typically preferred because it is the most common amino acid. Further, it Is frequently found in both buried and exposed positions (Creighton, The Proteins, Freeman Co., Chothia, J. Mol. Biol., 150:1 (1976)). If alanine substitution does not yield adequate amounts of variant, an isosteric amino acid can be used.

As defined herein, 109P1D4 variants, analogs or homologs, have the distinguishing attribute of having at least one epitope that is "cross reactive' with a 109P1D4 protein having an amino acid sequence of Figure 3. As used in this sentence, "cross reactive" means that an antibody or T cell that specifically binds to a 109P1D4 variant also specifically binds to a 109P1 D4 protein having an amino acid sequence set forth in Figure 3. A polypeptide ceases to be a variant of a protein shown in Figure 3, when It no longer contains any epitope capable of being recognized by an antibody or T cell that specifically binds to the starting 109P1D4 protein. Those skilled in the art understand that antibodies that recognize proteins bind to epitopes of varying size, and a grouping of the order of about four or five amino acds, contiguous or not is regarded as a typical number of amino acids in a minimal epitope. See, Nair etal., J. Immunol 2000 165(12): 6949-6955; Hebbes et Mol Immunol (1989) 26(9):865-73; Schwartz et J Immunol (1985) 135(4):2598-608.

Other classes of 10SP1D4-related protein variants share 70%, 75%, 80%, 85% or 90% or more similarity with an amino add sequence of Figure 3, or a fragment thereof. Another specific class of 109P1D4 protein variants or analogs comprises one or more of the 109P1D4 biological motifs described herein or presently known in the art Thus, encompassed by the present invention are analogs of 109P1D4 fragments (nucleic or amino add) that have altered functional (e.g.

immunogenic) properties relative to the starting fragment It is to be appreciated that motifs now or which become part of the art are to be applied to the nucleic or amino add sequences of Figure 2 or Figure 3.

As discussed herein, embodiments of the claimed invention indude polypeptides containing less than the full amino acid sequence of a 109P1D4 protein shown in Figure 2 or Figure 3. For example, representative embodiments of the invention comprise peptides/proteins having any 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more contiguous amino adds of a 00 109P1D4 protein shown in Figure 2 or Figure 3.

Moreover, representative embodiments of the invention disclosed herein include polypeptides consisting of about amino acid 1 to about amino acid 10 of a 109P1D4 protein shown in Figure 2 or Figure 3, polypeptides consisting of about S amino acid 10 to about amino add 20 of a 109PID4 protein shown in Figure 2 or Figure 3, polypeptides consisting of about amino acid 20 to about amino acid 30 of a 109P1D4 protein shown in Figure 2 or Figure 3, polypeptides consisting of about Samino add 30 to about amino add 40 of a 109P1 D4 protein shown in Figure 2 or Figure 3, polypeptides consisting of about amino add 40 to about amino add 50 of a 109P1D4 protein shown in Figure 2 or Figure 3, polypeptides consisting of about amino acid 50 to about amino add 60 of a 109P1D4 protein shown in Figure 2 or Figure 3, polypeptides consisting of about amino acid 60 to about amino acid 70 of a 109P1D4 protein shown in Figure 2 or Figure 3, polypeptides consisting of about Samino acid 70 to about amino acid 80 of a 109P1 D4 protein shown in Figure 2 or Figure 3, polypeptides consisting of about Samino add 80 to about amino acid 90 of a 109P1 D4 protein shown in Figure 2 or Figure 3, polypeptides consisting of about Samino acid 90 to about amino acid 100 of a 109P1 D4 protein shown in Figure 2 or Figure 3, etc. throughout the entirety of a 00 S109P1D4 amino acid sequence. Moreover, polypeptides consisting of about amino acid 1 (or 20 or 30 or 40 etc.) to about Samino acid 20, (or 130, or 140 or 150 etc.) of a 109P1D4 protein shown in Figure 2 or Figure 3 are embodiments of the invention. It.ls to be appreciated that the starting and stopping positions in this paragraph refer to the specified position as well as that position plus or minus 5 residues.

109P1D4-related proteins are generated using standard peptide synthesis technology or using chemical cleavage methods well known in the art Altematively, recombinant methods can be used to generate nucleic acid molecules that encode a 109P1D4-related protein. In one embodiment, nucleic add molecules provide a means to generate defined fragments of a 109P1 D4 protein (or variants, homologs or analogs thereof).

IIIA) Motif-bearing Protein Embodiments Additional illustrative embodiments of the invention disclosed herein include 109P1D4 polypeptides comprising the amino acid residues of one or more of the biological motifs contained within a 109P104 polypeptide sequence set forth in Figure 2 or Figure 3. Various motifs are known in the art, and a protein can be evaluated for the presence of such motifs by a number of publicly available Intemet sites (see, URL addresses: pfam.wusU.edul; searchlauncher.bcm.tmc.edulseqsearch/struo-predichtml; psortims.u-tokyo.ac.jp; cbs.dtu.dk/; ebi.ac.ukinterpro/scan.html; expasy.ch/tools/scnpsitl.html; Epimatrix and Epime r Brown University, brown.edu/ResearchB-HIVablepimatrixepimatix.htl; and BIMAS, bimas.dcrtnih.govl.).

Motif bearing subsequences of all 109P1D4 variant proteins are set forth and identified in Tables VIII-XXI and XXII-

XLIX

Table V sets forth several frequently occurring motifs based on pfam searches (see URL address pfam.wustl.edu/).

The columns of Table V list motif name abbreviation, percent identity found amongst the different member of the motif family, motif name or description and most common function; location information Is included if the motif is relevant for location.

Polypeptides comprising one or more of the 109P1D4 motifs discussed above are useful in elucidating the specific characteristics of a malignant phenotype in view of the observation that the 109P1D4 motifs discussed above are associated with growth dysregulation and because 109P1D4 is overexpressed in certain cancers (See, Table Casein kinase II, cAMP and camp-dependent protein kinase, and Protein Kinase C, for example, are enzymes known to be associated with the development of the malignant phenotype (see e.g. Chen et Lab Invest, 78(2): 165-174 (1998); Galddon et al., Endocrinology 136(10): 4331-4338 (1995); Hall et Nucleic Acids Research 24(6): 1119-1126 (1996); Peterziel et al., Oncogene 18(46): 6322-6329 (1999) and O'Brian, Oncol. Rep. 305-309 (1998)). Moreover, both glycosylation and 00 myristoylation are protein modifications also associated with cancer and cancer progression (see e.g. Dennis et al., Biochem.

SBiophys. Acta 1473(1):21-34 (1999); Raju etal., Exp. Cell Res. 235(1): 145-154 (1997)). Amidation is another protein N modification also associated with cancer and cancer progression (see e.g. Treston et al., J. Natl. Cancer Inst Monogr. (13): S169-175(1992)).

In another embodiment, proteins of the invention comprise one or more of the immunoreactive epitopes identified in accordance with art-accepted methods, such as the peptides set forth in Tables VIII-XXI and XXII-XLIX. CTL epitopes can be determined using specific algorithms to identify peptides within a 109P1D4 protein that are capable of optimally binding to specified HLA alleles Table IV; EpimatrixM and Epimer

M

Brown University, URL brown.edu/ResearchfB- HIVLablepimarix/epimatix.htm; and BIMAS, URL blmas.dcrtnih.gov/.) Moreover, processes for identifying peptides that have Ssufficient binding affinity for HLA molecules and which are correlated with being Immunogenic epitopes, are well known in the art, and are carried out without undue experimentation. In addition, processes for Identifying peptides that are immunogenic Sepitopes, are well known in the art, and are carried out without undue experimentation either In vitro or in vivo.

00 SAlso known in the art are principles for creating analogs of such epitopes in order to modulate immunogenidty. For example, one begins with an epitope that bears a CTL or HTL motif (see, the HLA Class I and HLA Class II motifs/supermotifs of Table IV). The epitope is analoged by substituting out an amino acid at one of the specified positions, and replacing it with another amino acid specified for that position. For example, on the basis of residues defined In Table IV, one can substitute out a deleterious residue in favor of any other residue, such as a preferred residue; substitute a lesspreferred residue with a preferred residue; or substitute an originally-occurring preferred residue with another preferred residue. Substitutions can occur at primary anchor positions or at other positions in a peptide; see,,e.g., Table IV.

A variety of references reflect the art regarding the identification and generation of epitopes in a protein of interest as well as analogs thereof. See, for example, WO 97/33602 to Chesnut et al.; Sette, Immunogenetics 1999 50(3-4): 201- 212; Sette et J. Immunol. 2001 166(2): 1389-1397; Sidney et al., Hum. Immunol. 1997 58(1): 12-20; Kondo etal., Immunogenetics 1997 45(4): 249-258; Sidney et J. Immunol. 1996 157(8): 3480-90; and Falk et Nature 351: 290-6 (1991); Hunt et Science 255:1261-3 (1992); Parker et al., J. Immunol. 149:3580-7 (1992); Parker et al, J. Immunol.

152:163-75 (1994)); Kast et al., 1994 152(8): 3904-12; Borras-Cuesta et al., Hum. Immunol. 2000 61(3): 266-278; Alexander etal., J. Immunol. 2000 164(3); 164(3): 1625-1633; Alexander et al., PMID: 7895164, UI: 95202582; O'Sullivan et J.

Immunol. 1991 147(8): 2663-2669; Alexander et al., Immunity 1994 751-761 and Alexander et al., Immunol. Res. 1998 18(2): 79-92.

Related embodiments of the invention include polypeptides comprising combinations of the different motifs set forth in Table VI, and/or, one or more of the predicted CTL epitopes of Tables VIII-XXI and XXII-XLIX, and/or, one or more of the predicted HTL epitopes of Tables XLV-XLIX, and/or, one or more of the T cell binding motifs known in the art Preferred embodiments contain no insertions, deletions or substitutions either within the motifs or within the intervening sequences of the polypeptides. In addition, embodiments which include a number of either N-terminal and/or C-terminal amino acid residues on either side of these motifs may be desirable (to, for example, indude a greater portion of the polypeptide architecture in which the motif is located). Typically, the number of N-terminal and/or C-terminal amino acid residues on either side of a motif is between about 1 to about 100 amino acid residues, preferably 5 to about 50 amino add residues.

109P1D4-related proteins are embodied in many forms, preferably in isolated form. A purified 109P1D4 protein molecule will be substantially free of other proteins or molecules that Impair the binding of 109P1D4 to antibody, T cell or other ligand. The nature and degree of isolation and purification will depend on the intended use. Embodiments of a 109P1D4related proteins include purified 109P1D4-related proteins and functional, soluble 109P1D4-related proteins. In one embodiment, a functional, soluble 109P1D4 protein or fragment thereof retains the ability to be bound by antibody, T cell or 00 other ligand.

The invention also provides 109P1D4 proteins comprising biologically active fragments of a 109P1D4 amino add C sequence shown in Figure 2 or Figure 3. Such proteins exhibit properties of the starting 109P1D4 protein, such as the ability to elidt the generation of antibodies that specifically bind an epitope associated with the starting 109P1D4 protein; to be C)bound by such antibodies; to elicit the activation of HTL or CTL; and/or, to be recognized by HTL or CTL that also specifically V) bind to the starting protein.

109P1D4-related polypeptides that contain particularly interesting structures can be predicted and/or identified using various analytical techniques well known in the art, including, for example, the methods of Chou-Fasman, Gamier-Robson, Kyte- SDoolittle, Eisenberg, Karplus-Schultz or Jameson-Wolf analysis, or based on immunogenicty. Fragments that contain such Sstructures are particularly useful in generating subunit-specific anti-109P1D4 antibodies or T cells or in identifying cellular factors S that bind to 109P1D4. For example, hydrophilidty profiles can be generated, and immunogenic peptide fragments identified, r using the method of Hopp, T.P. and Woods, KR., 1981, Proc. Nail. Acad. Sc. U.S.A. 78:3824-3828. Hydropathicity profiles can be generated, and immunogenic peptide fragments identified, using the method of Kyte, J. and Doolittle, 1982, J.

Mol. Biol. 157:105-132. Percent Accessible Residues profiles can be generated, and immunogenic peptide fragments identified, using the method of Janin 1979, Nature 277:491-492. Average Flexibility profiles can be generated, and immunogenic peptide fragments identified, using the method of Bhaskaran Ponnuswamy P.K, 1988, Int J. Pept. Protein Res. 32:242-255. Beta-turn profiles can be generated, and Immunogenic peptide fragments identified, using the method of Deleage, Roux 1987, Protein Engineering 1:289-294.

CTL epitopes can be determined using specific algorithms to identify peptides within a 109P1D4 protein that are capable of optimally binding to specified HLA alleles by using the SYFPEITHI site at World Wide Web URL syfpeithi.bmiheidelberg.conm; the listings in Table Epimatrix and Epimer", Brown University, URL (brown.edulResearch/TB- HIVLabeplmarixepmatrix.html); and BIMAS, URL blmas.dctnih.gov/). Illustrating this, peptide epitopes from 109P1D4 that are presented in the context of human MHC Class I molecules, HLA-A1, A2, A3, All, A24, B7 and B35 were predicted (see, Tables VIII-XXI, XXII-XLIX). Specifically, the complete amino acid sequence of the 109P1D4 protein and relevant portions of other variants, for HLA Class I predictions 9 flanking residues on either side of a point mutation or exon juction, and for HLA Class II predictions 14 flanking residues on either side of a point mutation or exon junction corresponding to that variant, were entered into the HLA Peptide Motif Search algorithm found in the Bioinformatics and Molecular Analysis Section (BIMAS) web site listed above; in addition to the site SYFPEITHI, at URL syfpelthi.bmiheldelberg.com/.

The HLA peptide motif search algorithm was developed by Dr. Ken Parker based on binding of specific peptide sequences in the groove of HLA Class I molecules, in particular HLA-A2 (see, Falk et al., Nature 351: 290-6 (1991); Hunt et al., Science 255:1261-3 (1992); Parker et J. Immunol. 149:3580-7 (1992); Parker et J. Immunol. 152:163-75 (1994)). This algorithm allows location and ranking of 8-mer, 9-mer, and 10-mer peptides from a complete protein sequence for predicted binding to HLA-A2 as well as numerous other HLA Class I molecules. Many HLA class I binding peptides are 10 or 11-mers. For example, for Class I HLA-A2, the epitopes preferably contain a leucine or methlonine at position 2 and a valine or leudne at the C-terminus (see, Parker et al., J. Immunol. 149:3580-7 (1992)).

Selected results of 109P1D4 predicted binding peptides are shown In Tables VIII-XXI and XXII-XIX herein. In Tables VIII- XXI and XXII-XLVl, selected candidates, 9-mers and 10-mers, for each family member are shown along with their location, the amino acid sequence of each specific peptide, and an estimated binding score. In Tables XLVI-XLIX, selected candidates, 15-mers, for each family member are shown along with their location, the amino acid sequence of each specific peptide, and an estimated binding score. The binding score corresponds to the estimated half time of dissodation of 00 complexes containing the peptide at 37oC at pH 6.5. Peptides with the highest binding score are predicted to be the most tightly bound to HLA Class I on the cell surface for the greatest period of time and thus represent the best immunogenic targets for T-cell recognition.

J) Actual binding of peptides to an HLA allele can be evaluated by stabilization of HLA expression on the antigenprocessing defective cell line T2 (see, Xue et Prostate 30:73-8 (1997) and Peshwa et al., Prostate 36:129-38 (1998)). Immunogenicity of specific peptides can be evaluated in vitro by stimulation of CD8+ cytotoxlc T lymphocytes (CTL) in the presence of antigen presenting cells such as dendritic cells.

It Is to be appreciated that every epitope predicted by the BIMAS site, Epimer T M and EpimatrixT M sites, or specified by the HLA class I or dass II motifs available in the art or which become part of the art such as set forth in Table IV (or Sdetermined using World Wide Web site URL syfpeithi.bmi-heidelberg.coml, or BIMAS, bimas.dcrtnih.gov/) are to be "applied' to a 109P1D4 protein in accordance with the invention. As used in this context 'applied' means that a 109P1D4 protein is Q0 evaluated, visually or by computer-based patterns finding methods, as appreciated by those of skill in the relevant art.

O Every subsequence of a 109P1D4 protein of 8, 9, 10, or 11 amino acid residues that bears an HLA Class I motif, or a subsequence of 9 or more amino acid residues that bear an HLA Class II motif are within the scope of the invention.

III.B.) Expression of 109PiD4-related Proteins In an embodiment described in the examples that follow, 109P1D4 can be conveniently expressed in cells (such as 293T cells) transfected with a commercially available expression vector such as a CMV-driven expression vector encoding 109P1 D4 with a C-terminal 6XHis and MYC tag (pcDNA3.1/mycHIS, Invitrogen or Tag5, GenHunter Corporation, Nashville TN). The Tag5 vector provides an IgGK secretion signal that can be used to facilitate the production of a secreted 109P1D4 protein in transfected cells. The secreted HIS-tagged 109P1D4 in the culture media can be purified, using a nickel column using standard techniques.

III.C.) Modifications of 109P1 D4-related Proteins Modifications of 109P1D4-related proteins such as covalent modifications are induded within the scope of this invention. One type of covalent modification includes reacting targeted amino acid residues of a 109P1D4 polypeptide with an organic derivatizing agent that is capable of reacting with selected side chains or the N- or C- terminal residues of a 109P104 protein. Another type of covalent modification of a 109P1D4 polypeptide included within the scope of this invention comprises altering the native glycosylation pattern of a protein of the invention. Another type of covalent modification of 109P1D4 comprises Unking a 109P1D4 polypeptide to one of a variety of nonprotelnaceous polymers, polyethylene glycol (PEG), polypropylene glycol, or polyoxyalkylenes, in the manner set forth in U.S. Patent Nos. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192 or 4,179,337.

The 109P1 D4-related proteins of the present invention can also be modified to form a chimeric molecule comprising 109P1D4 fused to another, heterologous polypeptide or amino acid sequence. Such a chimeric molecule can be synthesized chemically or recombinantly. A chimeric molecule can have a protein of the invention fused to another tumorassociated antigen or fragment thereof. Altematively, a protein In accordance with the invention can comprise a fusion of fragments of a 109P1D4 sequence (amino or nucleic acid) such that a molecule Is created that is not, through its length, directly homologous to the amino or nucleic add sequences shown In Figure 2 or Figure 3. Such a chimeric molecule can comprise multiples of the same subsequence of 109P1D4. Achimeric molecule can comprise a fusion of a 109P1D4-related protein with a polyhistidine epitope tag, which provides an epitope to which immobilized nickel can selectively bind, with cytokines or with growth factors. The epitope tag is generally placed at the amino- or carboxyl- terminus of a 109P1D4 0 0 protein. In an alternative embodiment the chimeric molecule can comprise a fusion of a 109P1D4-related protein with an immunoglobulin or a particular region of an immunoglobulin. For a bivalent form of the chimeric molecule (also referred to as an "immunoadhesin"), such a fusion could be to the Fc region of an IgG molecule. The Ig fusions preferably include the substitution of a soluble (transmembrane domain deleted or inactivated) form of a 109P1D4 polypeptide in place of at least one variable region within an Ig molecule. In a preferred embodiment, the immunoglobulin fusion includes the hinge, CH2 Sand CH3, or the hinge, CHI, CH2 and CH3 regions of an IgGI molecule. For the production of immunoglobulin fusions see, U.S. Patent No. 5,428,130 Issued June 27, 1995.

111.D.) Uses of 109P1D4-related Proteins SThe proteins of the invention have a number of different specific uses. As 109P1D4 is highly expressed in prostate and other cancers, 109P1D4-related proteins are used in methods that assess the status of 109P1D4 gene products in 0normal versus cancerous tissues, thereby elucidating the malignant phenotype. Typically, polypeptides from specific regions O of a 109P1D4 protein are used to assess the presence of perturbations (such as deletions, insertions, point mutations etc.) in Sthose regions (such as regions containing one or more motifs). Exemplary assays utilize antibodies or T cells targeting 109P1 D4-related proteins comprising the amino add residues of one or more of the biological motifs contained within a 109P1D4 polypeptide sequence in order to evaluate the characteristics of this region in normal versus cancerous tissues or to elicit an immune response to the epitope. Alternatively, 109P1D4-related proteins that contain the amino add residues of one or more of the biological motifs in a 109P1D4 protein are used to screen for factors that interact with that region of 109P1D4.

109P1D4 protein fragments/subsequences are particularly useful in generating and characterizing domain-specific antibodies antibodies recognizing an extracellular or intracellular epitope of a 109P1D4 protein), for identifying agents or cellular factors that bind to 109P1D4 or a particular structural domain thereof, and in various therapeutic and diagnostic contexts, including but not limited to diagnostic assays, cancer vaccines and methods of preparing such vaccines.

Proteins encoded by the 109P1D4 genes, or by analogs, homologs or fragments thereof, have a variety of uses, including but not limited to generating antibodies and in methods for identifying ligands and other agents and cellular constituents that bind to a 109P1D4 gene product Antibodies raised against a 109P1D4 protein or fragment thereof are useful in diagnostic and prognostic assays, and imaging methodologies in the management of human cancers characterized by expression of 109P1D4 protein, such as those listed in Table I. Such antibodies can be expressed intracellularly and used in methods of treating patients with such cancers. 109P1D4-related nucleic acids or proteins are also used in generating HTL or CTL responses.

Various immunological assays useful for the detection of 109P1D4 proteins are used, Including but not limited to various types of radiolmmunoassays, enzymelinked nmmunosorbent assays (EUSA), enzyme-linked Immunofluorescent assays (ELIFA), immunocytochemical methods, and the like. Antibodies can be labeled and used as immunological Imaging reagents capable of detecting 109P1D4-expressing cells in radioscintigraphic imaging methods). 109P1D4 proteins are also particularly useful in generating cancer vacines, as further described herein.

IV.) 109P1D4Antibodies Another aspect of the invention provides antibodies that bind to 109P1D4-related proteins. Preferred antibodies specifically bind to a 109P D4-related protein and do not bind (or bind weakly) to peptides or proteins that are not 109P1D4related proteins under physological conditions. In this context, examples of physiological conditions include: 1) phosphate buffered saline; 2) Tris-buffered saline containing 25mM Tris and 150 mM NaCI; or normal saline NaCI); 4) animal serum 00 such as human serum; or, 5) a combination of any of 1) through these reactions preferably taking place at pH 7.5, alternatively in a range of pH 7.0 to 8.0, or alternatively in a range of pH 6.5 to 8.5; also, these reactions taking place at a temperature between 4°C to 370C. For example, antibodies that bind 109P1 D4 can bind 109P1 D4-related proteins such as the homologs or analogs thereof.

I 09P1 D4 antibodies of the invention are particularly useful in cancer (see, Table I) diagnostic and prognostic Sassays, and Imaging methodologies. Similarly, such antibodies are useful in the treatment, diagnosis, and/or prognosis of other cancers, to the extent 109P1 D4 is also expressed or overexpressed in these other cancers. Moreover, intracellularly expressed antibodies single chain antibodies) are therapeutically useful in treating cancers in which the expression of S109P1D4 is involved, such as advanced or metastatic prostate cancers.

SThe invention also provides various immunological assays useful for the detection and quantification of 109P1D4 and mutant 109P1D4-related proteins. Such assays can comprise one or more 109P1 D4 antibodies capable of recognizing and 0 binding a 109P1 D4-related protein, as appropriate. These assays are performed within various immunological assay formats well 00 0 known in the art, including but not limited to various types of radioimmunoassays, enzyme-linked immunosorbent assays (EUSA), enzyme-linked Immunofluorescent assays (ELIFA), and the like.

Immunological non-antibody assays of the invention also comprise T cell immunogenicity assays (inhibitory or stimulatory) as well as major histocompatibility complex (MHC) binding assays.

In addition, immunological imaging methods capable of detecting prostate cancer and other cancers expressing 109P1D4 are also provided by the invention, induding but not limited to radioscintigraphic imaging methods using labeled 109PiD4 antibodies. Such assays are dinically useful in the detection, monitoring, and prognosis of 109P1D4 expressing cancers such as prostate cancer.

109P1D4 antibodies are also used in methods for purifying a 109P1D4-related protein and for isolating 109P1D4 homologues and related molecules. For example, a method of purifying a 109P1D4-related protein comprises incubating a 109P1D4 antibody, which has been coupled to a solid matrix, with a lysate or other solution containing a 109P104-related protein under conditions that permit the 109P1D 4 antibody to bind to the 109P1 D4-related protein; washing the solid matrix to eliminate impurities; and eluting the 109P1D4-lated protein from the coupled antibody. Other uses of 109P1D4 antibodies in accordance with the invention include generating anti-idiotypic antibodies that mimic a 109P1D4 protein.

Various methods for the preparation of antibodies are well known in the art For example, antibodies can be prepared by immunizing a suitable mammalian host using a 109P1D4-related protein, peptide, or fragment, in isolated or immunoconjugated form (Antibodies: A Laboratory Manual, CSH Press, Eds., Harlow, and Lane (1988); Harlow, Antibodies, Cold Spring Harbor Press, NY (1989)). In addition, fusion proteins of 109P1D4 can also be used, such as a 109P1D4 GST-fusion protein. In a particular embodiment, a GST fusion protein comprising al or most of the amino add sequence of Figure 2 or Figure 3 is produced, then used as an immunogen to generate appropriate antibodies. In another embodiment, a 109P1D4-related protein Is synthesized and used as an immunogen.

In addition, naked DNA immunization techniques known in the art are used (with or without purified 109P1 D4-related protein or 109P1D4 expressing cells) to generate an immune response to the encoded immunogen (for review, see Donnelly et at, 1997, Ann. Rev. Immunol. 15: 617-648).

The amino add sequence of a 109P1D4 protein as shown in Figure 2 or Figure 3 can be analyzed to select specific regions of the 109P1D4 protein for generating antibodies. For example, hydrophobicity and hydrophilicty analyses of a 109P1D4 amino acid sequence are used to identify hydrophlic regions in the 109P1D4 structure. Regions of a 109P1D4 protein that show immunogenic structure, as well as other regions and domains, can readily be Identified using various other methods known in the art, such as Chou-Fasman, Gamier-Robson, Kyte-Doolittle, Eisenberg, Karplus-Schultz or Jameson-Wolf analysis. Hydrophilidty 00 profiles can be generated using the method of Hopp, T.P. and Woods, 1981, Proc. Natl. Acad. Sci. U.S.A. 78:3824- 3828. Hydropathicity profiles can be generated using the method of Kyte, J. and Doolittle, 1982, J. Mol. Biol. 157:105r 132. Percent Accessible Residues profiles can be generated using the method of Janin 1979, Nature 277:491-492.

i Average Flexibility profiles can be generated using the method of Bhaskaran Ponnuswamy P.K, 1988, Int J. Pept C)Protein Res. 32:242-255. Beta-turn profiles can be generated using the method of Deleage, Roux 1987, Protein ^f Engineering 1:289-294. Thus, each region identified by any of these programs or methods is within the scope of the present invention. Methods for the generation of 109P1D4 antibodies are further illustrated by way of the examples provided herein.

Methods for preparing a protein or polypeptide for use as an immunogen are well known in the art Also well known in the art are Smethods for preparing immunogenic conjugates of a protein with a carrer, such as BSA, KLH or other carrier protein. In some Scircumstances, direct conjugation using, for example, carbodiimide reagents are used; in other instances linking reagents such as those supplied by Pierce Chemical Co., Rockford, IL are effective. Administration of a 109P1 D4 immunogen is often conducted C by injection over a suitable time period and with use of a suitable adjuvant, as is understood in the art During the immunization 00 Sschedule, liters of antibodies can be taken to determine adequacy of antibody formation.

S109P1D4 monoclonal antibodies can be produced by various means well known in the art For example, immortalized cell lines that secrete a desired monodonal antibody are prepared using the standard hybridoma technology of Kohler and Milstein or modifications that immortalize antibody-producing B cells, as is generally known. Immortalized cell lines that secrete the desired antibodies are screened by immunoassay in which the antigen is a 109P1D4-related protein. When the appropriate Immortalized cell culture is Identified, the cells can be expanded and antibodies produced either from in vito cultures or from ascites fluid.

The antibodies or fragments of the invention can also be produced, by recombinant means. Regions that bind specifically to the desired regions of a 109P1D4 protein can also be produced in the context of chimeric or complementaritydetermining region (CDR) grafted antibodies of multiple species origin. Humanized or human 109P1 D4 antibodies can also be produced, and are preferred for use in therapeutic contexts. Methods for humanizing murine and other non-human antibodies, by substituting one or more of the non-human antibody CDRs for corresponding human antibody sequences, are well known (see for example, Jones et al., 1986, Nature 321: 522-525; Riechmann etal., 1988, Nature 332: 323-327; Verhoeyen et al., 1988, Science 239: 1534-1536). See also, Carter eta., 1993, Proc Nal. Acad. Sc. USA 89:4285 and Sims et al., 1993, J. Immunol. 151: 2296.

Methods for producing fully human monodonal antibodies include phage display and transgenic methods (for review, see Vaughan et al., 1998, Nature Biotechnology 16: 535-539). Fully human 109P1D4 monoconal antibodies can be generated using cloning technologies employing large human Ig gene combinatorial libraries phage display) (Griffiths and Hoogenboom, Building an in vito immune system: human antibodies from phage display libraries. In: Protein Engineering of Antibody Molecules for Prophylactic and Therapeutic Applications in Man, Clark, M. Nottingham Academic, pp 45-64 (1993); Burton and Barbas, Human Antibodies from combinatorial libraries. Id., pp 65-82). Fully human 109P1D4 monoconal antibodies can also be produced using transgenic mice engineered to contain human immunoglobulin gene loci as described in PCT Patent Application WO98/24893, Kucherlapati and Jakobovits et al., published December 3,1997 (see also, Jakobovits, 1998, Exp. Opin. Invest Drugs 607-614; U.S. patents 6,162963 issued 19 December 2000; 6,150,584 issued 12 November 2000; and, 6,114598 Issued 5 September 2000). This method avoids the in vito manipulation required with phage display technology and efficiently produces high affinity authentic human antibodies.

Reactivity of 109P1D4 antibodies with a 109P1D4-related protein can be established by a number of well known means, including Western blot, immunoprecpitation, ELISA, and FACS analyses using, as appropriate, 109P1D4-related proteins, 109P1D4-expressing cells or extracts thereof. A 109P1D4 antibody or fragment thereof can be labeled with a detectable marker or conjugated to a second molecule. Suitable detectable markers Include, but are not limited to, a 00 radioisotope, a fluorescent compound, a bioluminescent compound, chemiluminescent compound, a metal chelator or an enzyme. Further, bi-specific antibodies specific for two or more 109P1 D4 epitopes are generated using methods generally CK known in the art Homodimeric antibodies can also be generated by cross-linking techniques known in the art Wolff at al., Cancer Res. 53: 2560-2565).

n 109PD4 Cellular Immune Responses The mechanism by which T cells recognize antigens has been delineated. Efficacious peptide epitope vaccine compositions of the invention induce a therapeutic or prophylactic immune responses in very broad segments of the world- Swide population. For an understanding of the value and efficacy of compositions of the invention that induce cellular immune 0responses, a brief review of immunology-related technology is provided.

SA complex of an HLA molecule and a peptidic antigen acts as the ligand recognized by HLA-restricted T cells CN (Buus, S. et al., Cell 47:1071, 1986; Babbitt, B. P. at al., Nature 317:359,1985; Townsend, A. and Bodmer, Annu. Rev.

00 Immunol. 7:601, 1989; Germaln, R Annu. Rev. Immunol. 11:403,1993). Through the study of single amino acid Ssubstituted antigen analogs and the sequencing of endogenously bound, naturally processed peptides, critical residues that correspond to motifs required for specific binding to HLA antigen molecules have been identified and are set forth in Table IV (see also, Southwood, et al., J. Immunol. 160:3363,1998; Rammensee, et al., Immunogenetics 41:178,1995; Rammensee eta., SYFPEITHI, access via World Wide Web at URL (134.2.96.221/scripts.hlaserver.dl/home.htm); Sette, A.

and Sidney, J. Curr. Opin. Immunol. 10:478,1998; Engelhard, V. Cur. Opin. Immunol. 6:13,1994; Sette, A. and Grey, H.

Curr. Opin. Immunol. 4:79,1992; Sinigaglia, F. and Hammer, J. Curr. Biol. 6:52, 1994; Ruppert et al., Cel 74:929-937, 1993; Kondo e al., J. Immunol. 155:4307-4312,1995; Sidney et J. Immunol. 157:3480-3490,1996; Sidney et al., Human Immunol. 45:79-93, 1996; Sette, A. and Sidney, J. Immunogenetics 1999 Nov; 50(3-4):201-12, Review).

Furthermore, x-ray crystallographic analyses of HLA-peptide complexes have revealed pockets within the peptide binding cleft/groove of HLA molecules which accommodate, in an allele-specific mode, residues borne by peptide igands; these residues in turn determine the HLA binding capacity of the peptides in which they are present (See, Madden, D.R. Annu. Rev. Immunol. 13:587,1995; Smith, et Immunity4:203, 1996; Fremont et Immunity 8:305, 1998; Stem et al., Structure 2:245,1994; Jones, E.Y. Cufr. Op/n. Immunol. 9:75, 1997; Brown, J. H. etal., Nature 364:33, 1993; Guo, H. C.

etal., Proc. Natl. Acad. Sc. USA 90:8053,1993; Guo, H. C. eta/., Nature 360:364,1992; Silver, M. L etal., Nature 360:367, 1992; Matsumura, M. et Science 257:927,1992; Madden etal., Ce/l70:1035, 1992; Fremont, D. H. et al., Science 257:919, 1992; Saper, M. Bjorkman, P. J. and Wiley, D. J. Mol. Biol. 219:277, 1991.) Accordingly, the definition of class I and dass II allele-specific HLA binding motifs, or class I or class II supermotifs allows identification of regions within a protein that are correlated with binding to particular HLA antigen(s).

Thus, by a process of HLA motif identification, candidates for epitope-based vaccines have been identified; such candidates can be further evaluated by HLA-peptide binding assays to determine binding affinity and/or the time period of association of the epitope and its corresponding HLA molecule. Additional confirmatory work can be performed to select, amongst these vaccine candidates, epitopes with preferred characteristics in terms of population coverage, and/or immunogenicity.

Various strategies can be utilized to evaluate cellular immunogenicity, including: 1) Evaluation of primary T cell cultures from normal individuals (see, Wentworth, P. A. et al., Mol. Immunol.

32:603, 1995; Cells, E. et al., Proc. Natl. Acad. Sci. USA 91:2105, 1994; Tsai, V. et J. Immunol. 158:1796,1997; Kawashima, I. et al., Human Immunol. 59:1, 1998). This procedure involves the stimulation of peripheral blood lymphocytes (PBL) from normal subjects with a test peptide in the presence of antigen presenting cells in vitro over a period of several 00 weeks. Tcells specific for the peptide become activated during this time and are detected using, a lymphokine- or 1 Cr-release assay involving peptide sensitized target cells.

N 2) Immunization of HLA transgenic mice (see, Wentworth, P. A. et al., J. Immunol. 2697, 1996; Wentworth, P.

A. et al., Int. Immunol. 8:651, 1996; Alexander, J. et al., J. Immunol. 159:4753, 1997). For example, in such methods peptides in incomplete Freund's adjuvant are administered subcutaneously to HLA transgenic mice. Several weeks following Simmunization, splenocytes are removed and cultured in viro in the presence of test peptide for approximately one week.

Peptide-specific T cells are detected using, a 5 1 Cr-release assay involving peptide sensitized target cells and target cells expressing endogenously generated antigen.

C3) Demonstration of recall T cell responses from immune individuals who have been either effectively vaccinated and/or from chronically ill patients (see, Rehermann, B. et al., J. Exp. Med. 181:1047, 1995; Doolan, D. L. ef a., Immunity 7:97, 1997; Bertoni, R. et al., J. Clin. Invest. 100:503, 1997; Threlkeld, S. C. et al., J. Immunol. 159:1648, 1997; O Diepolder, H. M. etal., J. Virol. 71:6011, 1997). Accordingly, recall responses are detected by culturing PBL from subjects Sthat have been exposed to the antigen due to disease and thus have generated an immune response "naturally', or from C patients who were vaccinated against the antigen. PBL from subjects are cultured in vitro for 1-2 weeks in the presence of test peptide plus antigen presenting cells (APC) to allow activation of memory" T cells, as compared to "naive" T cells. At the end of the culture period, T cell activity is detected using assays Including 51 Cr release involving peptide-sensitized targets, T cell proliferation, or lymphokine release.

VI,) 109PD4 Transgenic Animals Nucleic acids that encode a 109P D4-related protein can also be used to generate either transgenic animals or 'knock out" animals that, in turn, are useful in the development and screening of therapeutically useful reagents. In accordance with established techniques, cDNA encoding 109PiD4 can be used to done genomic DNA that encodes 109P1D4. The cloned genomic sequences can then be used to generate transgenic animals containing cells that express DNA that encode 109P1D4. Methods for generating transgenic animals, particularly animals such as mice or rats, have become conventional in the art and are described, for example, in U.S. Patent Nos. 4,736,866 issued 12 April 1988, and 4,870,009 issued 26 September 1989. Typically, particular cells would be targeted for 109P1D4 transgene incorporation with tissue-specific enhancers.

Transgenic animals that include a copy of a transgene encoding 109P1D4 can be used to examine the effect of increased expression of DNA that encodes 109P1D4. Such animals can be used as tester animals for reagents thought to confer protection from, for example, pathological conditions assodated with its overexpression. In accordance with this aspect of the Invention, an animal is treated with a reagent and a reduced Incidence of a pathological condition, compared to untreated animals that bear the transgene, would indicate a potential therapeutic intervention for the pathological condition.

Altematively, non-human homologues of 109P1D4 can be used to construct a 109P1D4 "knock out" animal that has a defective or altered gene encoding 109P1D4 as a result of homologous recombination between the endogenous gene encoding 109P1D4 and altered genomic DNA encoding 109P1 D4 Introduced into an embryonic cell of the animal. For example, cDNA that encodes 109P1D4 can be used to done genomic DNA encoding 109P1 D4 in accordance with established techniques. A portion of the genomic DNA encoding 109P1D4 can be deleted or replaced with another gene, such as a gene encoding a selectable marker that can be used to monitor Integration. Typically, several kilobases of unaltered flanking DNA (both at the 5' and 3' ends) are included in the vector (see, Thomas and Capecchi, Cell. 5:503 (1987) for a description of homologous recombination vectors). The vector is introduced into an embryonic stem cell line by electroporation) and cells in which the Introduced DNA has homologously recombined with the endogenous DNA 0 are selected (see, Li et al., ell, 69:915 (1992)). The selected cells are then injected into a blastocyst of an animal O a mouse or rat) to form aggregation chimeras (see, Bradley, in Teratocarcinomas and Embryonic Stem Cells: A Practical Approach, E. J. Robertson, ed. (IRL, Oxford, 1987), pp. 113-152). A chimeric embryo can then be implanted into a suitable pseudopregnant female foster animal, and the embryo brought to term to create a 'knock out" animal. Progeny Sharboring the homologously recombined DNA in their germ cells can be identified by standard techniques and used to breed l animals in which all cells of the animal contain the homologously recombined DNA Knock out animals can be characterized, Sfor example, for their ability to defend against certain pathological conditions or for their development of pathological conditions due to absence of a 109P1 D4 polypeptide.

0 VIL Methods forthe Detection of 109P1D4 1 Another aspect of the present invention relates to methods for detecting 109P1D4 polynucleolides and 109P1D4- Srelated proteins, as well as methods for identifying a cell that expresses 109P1D4. The expression profile of 109P1D4 makes it 00 00 a diagnostic marker for metastasized disease. Accordingly, the status of 109P1 D4 gene products provides information useful Sfor predicting a variety of factors including susceptibility to advanced stage disease, rate of progression, and/or tumor aggressiveness. As discussed in detail herein, the status of 109P1D4 gene products in patient samples can be analyzed by a variety protocols that are well known in the art including immunohistochemical analysis, the variety of Northem blotting techniques Including in situ hybridization, RT-PCR analysis (for example on laser capture micro-dssected samples), Western blot analysis and tissue array analysis.

More particularly, the invention provides assays for the detection of 109P1D4 polynudeotides in a biological sample, such as serum, bone, prostate, and other tissues, urine, semen, cell preparations, and the like. Detectable 109P1D4 polynudeotides include, for example, a 109P1D4 gene or fragment thereof, 109P1D4 mRNA, altemative splice variant 109P1D4 mRNAs, and recombinant DNA or RNA molecules that contain a 109P1D4 polynudeotide. A number of methods for amplifying and/or detecting the presence of 109P1D4 polynudeotides are well known in the art and can be employed in the practice of this aspect of the invention.

In one embodiment, a method for detecting a 109P1D4 mRNA in a biological sample comprises producing cDNA from the sample by reverse transcription using at least one primer; amplifying the cDNA so produced using a 109P1D4 polynucleotides as sense and antisense primers to amplify 109P1D4 cDNAs therein; and detecting the presence of the amplified 109P1D4 cONA. Optionally, the sequence of the amplified 109P1D4 cDNA can be determined.

In another embodiment, a method of detecting a 109P1D4 gene in a biological sample comprises first isolating genomic DNA from the sample; amplifying the isolated genomic DNA using 109P1D4 polynucleotides as sense and antisense primers; and detecting the presence of the amplified 109P1D4 gene. Any number of appropriate sense and antisense probe combinations can be designed from a 109P1D4 nudeotide sequence (see, Figure 2) and used for this purpose.

The invention also provides assays for detecting the presence of a 109P1 D4 protein in a tissue or other biological sample such as serum, semen, bone, prostate, urine, cell preparations, and the like. Methods for detecting a 109P1D4-related protein are also well known and indude, for example, Immunoprecipitation, Immunohistochemical analysis, Western blot analysis, molecular binding assays, ELISA, ELIFA and the like. For example, a method of detecting the presence of a 109P1 D4-related protein in a biological sample comprises first contacting the sample with a 109P1D4 antibody, a 109P1D4-reactive fragment thereof, or a recombinant protein containing an antigen-binding region of a 109P1 D4 antibody; and then detecting the binding of 109P1D4-related protein in the sample.

Methods for identifying a cell that expresses 109P1D4 are also within the scope of the invention. In one embodiment, 00 an assay for identifying a cell that expresses a 109P1D4 gene comprises detecting the presence of 109P1D4 mRNA in the cell.

Methods for the detection of particular mRNAs in cells are well known and indude, for example, hybridization assays using complementary DNA probes (such as in sftu hybridization usIng labeled 109P1D4 nboprobes, Northern blot and related techniques) and various nucleic acid amplification assays (such as RT-PCR using complementary primers specific for 109P1 D4, and other amplification type detection methods, such as, for example, branched DNA, SISBA, TMA and the like). Alternatively, an assay for identifying a cell that expresses a 109P1D4 gene comprises detecting the presence of 109P1 D4-related protein in the cell or secreted by the cell. Various methods for the detection of proteins are well known in the art and are employed for the detection of 109P1D4-related proteins and cells that express 109P1 D4-related proteins.

109P1D4 expression analysis Is also useful as a tool for identifying and evaluating agents that modulate 109PID4 gene expression. For example, 109P1 D4 expression is significantly upregulated in prostate cancer, and Is expressed In cancers of _the tissues listed in Table I. Identification of a molecule or biological agent that inhibits 109P1D4 expression or overrexpression in cancer cells Is of therapeutic value. For example, such an agent can be identified by using a screen that 00 quantifies 109P1D4 expression by RT-PCR, nucleic acid hybridization or antibody binding.

ViII.) Methods for Monitoring the Status of 109P1D4-related Genes and Their Products Oncogenesis is known to be a multistep process where cellular growth becomes progressively dysregulated and cells progress from a normal physiological state to precancerous and then cancerous states (see, Alers et al., Lab Invest 77(5): 437-438 (1997) and Isaacs et al., Cancer Surv. 23:19-32 (1995)). In this context, examining a biological sample for evidence of dysregulated cell growth (such as aberrant 109P1 D4 expression in cancers) allows for early detection of such aberrant physiology, before a pathologic state such as cancer has progressed to a stage that therapeutic options are more limited and or the prognosis is worse. In such examinations, the status of 109P1 D4 In a biological sample of interest can be compared, for example, to the status of 109P1D4 in a corresponding normal sample a sample from that individual or alternatively another Individual that is not affected by a pathology). An alteration In the status of 109P1D4 In the biological sample (as compared to the normal sample) provides evidence of dysregulated cellular growth. In addition to using a biological sample that is not affected by a pathology as a normal sample, one can also use a predetermined normative value such as a predetermined normal level of mRNA expression (see, Grever et al., J. Comp. Neurol. 1996 Dec 9; 376(2): 306-14 and U.S. Patent No. 5,837,501) to compare 109P1D4 status in a sample.

The term "status' in this context is used according to its art accepted meaning and refers to the condition or state of a gene and its products. Typically, sidlled artisans use a number of parameters to evaluate the condition or state of a gene and its products. These include, but are not limited to the location of expressed gene products (including the location of 109P1 D4 expressing cells) as well as the level, and biological activity of expressed gene products (such as 109P1 D4 mRNA, polynucleolides and polypeptides). Typically, an alteration in the status of 109P1D4 comprises a change in the location of 109P1 D4 and/or 109P1 D4 expressing cells and/or an Increase In 109P1 D4 mRNA and/or protein expression.

109P1D4 status in a sample can be analyzed by a number of means well known In the art, Induding without limitation, Immunohistochemical analysis, In situ hybridization, RT-PCR analysis on laser capture mlao-dissected samples, Western blot analysis, and tissue array analysis. Typical protocols for evauaing the status of a 109P1 D4 gene and gene products are found, for example In Ausubel etal. eds., 1995, Current Protocols In Molecular Biology, Units 2 (Northern Blotting), 4 (Southern Blotting), 15 (Immunoblotting) and 18 (PCR Analysis). Thus, the status of 109P1D4 in a biological sample Is evaluated by various methods utilized by skilled artisans Including, but not limited to genomic Southern analysis (to examine, for example perturbations in a 109P1 D4 gene), Northern analysis and/or PCR analysis of 109P1 D4 mRNA (to examine, for example alterations In the polynudeotide sequences or expression levels of 109P1 D4 mRNAs), and, Western and/or 44 0 immunohistochemical analysis (to examine, for example alterations in polypeptide sequences, alterations in polypeptide localization within a sample, alterations in expression levels of 109P1D4 proteins and/or associations of 109P1D4 proteins Swith polypeptide binding partners). Detectable 109P1D4 polynudeotides indude, for example, a 109P1D4 gene or fragment thereof, 109P1D4 mRNA, alterative splice variants, 109P1D4 mRNAs, and recombinant DNA or RNA molecules containing a 109P1D4 polynudeotide.

SThe expression profile of 109P1 D4 makes it a diagnostic marker for local and/or metastasized disease, and 0provides information on the growth or oncogenic potential of a biological sample. In particular, the status of 109P1D4 provides information useful for predicting susceptibility to particular disease stages, progression, and/or tumor aggressiveness. The 0 invention provides methods and assays for determining 109P1D4 status and diagnosing cancers that express 109P1D4, such as Scancers of the tissues listed in Table I. For example, because 109P1D4 mRNA is so highly expressed in prostate and other C cancers relative to normal prostate tissue, assays that evaluate the levels of 109P1D4 mRNA ranscripts or proteins in a biological Ssample can be used to diagnose a disease associated with 109P1D4 dysregulation, and can provide prognostic information useful 00 in defining appropriate therapeutic options.

The expression status of 109P1D4 provides Infonation including the presence, stage and location of dysplastic, precancerous and cancerous cells, predicting susceptibility to various stages of disease, and/or for gauging tumor aggressiveness. Moreover, the expression profile makes it useful as an imaging reagent for metastasized disease.

Consequently, an aspect of the invention is directed to the various molecular prognostic and diagnostic methods for examining the status of 109P1 D4 in biological samples such as those from individuals suffering from, or suspected of suffering from a pathology characterized by dysregulated cellular growth, such as cancer.

As described above, the status of 109P1D4 In a biological sample can be examined by a number of well-known procedures in the art For example, the status of 109P1D4 in a biological sample taken from a specific location in the body can be examined by evaluating the sample for the presence or absence of 109P1D4 expressing cells those that express 109P1D4 mRNAs or proteins). This examination can provide evidence of dysregulated cellular growth, for example, when 109P1D4-expressing cells are found in a biological sample that does not normally contain such cells (such as a lymph node), because such alterations in the status of 109P1D4 in a biological sample are often associated with dysregulated cellular growth. Specifically, one indicator of dysregulated cellular growth is the metastases of cancer cells from an organ of origin (such as the prostate) to a different area of the body (such as a lymph node). In this context, evidence of dysregulated cellular growth is important for example because occult lymph node metastases can be detected in a substantial proportion of patients with prostate cancer, and such metastases are associated with known predictors of disease progression (see, Murphy et aL, Prostate 42(4): 315-317 (2000);Su etal., Semin. Surg. Oncol. 18(1): 17-28 (2000) and Freeman et J Urol 1995 Aug 154(2 Pt 1):474-8).

In one aspect, the invention provides methods for monitoring 109P1D4 gene products by determining the status of 109P1D4 gene products expressed by cells from an individual suspected of having a disease associated with dysregulated cell growth (such as hyperplasia or cancer) and then comparing the status so determined to the status of 109P1 D4 gene products in a corresponding normal sample. The presence of aberrant 109P1D4 gene products in the test sample relative to the normal sample provides an indication of the presence of dysregulated cell growth within the cells of the individual.

In another aspect, the invention provides assays useful in determining the presence of cancer in an individual, comprising detecting a significant Increase In 109P1D4 mRNA or protein expression in a test cell or tissue sample relative to expression levels in the corresponding normal cell or tissue. The presence of 109P1 D4 mRNA can, for example, be evaluated in tissues including but not limited to those listed In Table I. The presence of significant 109P1D4 expression in any of these tissues is useful to indicate the emergence, presence and/or severity of a cancer, since the corresponding 00 normal tissues do not express 109P1D4 mRNA or express it at lower levels.

O In a related embodiment 109P1D4 status is determined at the protein level rather than at the nudeic add level. For 1 example, such a method comprises determining the level of 109P1 D4 protein expressed by cells in a test tissue sample and comparing the level so determined to the level of 109P1D4 expressed in a corresponding normal sample. In one embodiment Vj) the presence of 109P1D4 protein is evaluated, for example, using immunohistochemical methods. 109P1D4 antibodies or binding partners capable of detecting 109P1D4 protein expression are used in a variety of assay formats well known in the art for this purpose.

In a further embodiment one can evaluate the status of 109P1D4 nudeotide and amino acid sequences in a biological Ssample in order to identify perturbations in the structure of these molecules. These perturbations can indude insertions, deletions, Ssubstitutions and the like. Such evaluations are useful because perturbations in the nudeotide and amino add sequences are Sobserved in a large number of proteins associated with a growth dysregulated phenotype (see, Marrogl et al., 1999, J.

CN Cutan. Pathol. 26(8):369-378). For example, a mutation in the sequence of 109P1D4 may be indicative of the presence or 00 promotion of a tumor. Such assays therefore have diagnostic and predictive value where a mutation in 109P1D4 indicates a 0 potential loss of function or increase in tumor growth.

A wide variety of assays for observing pertrbatons in nudeotide and amino add sequences are well known in the art For example, the size and structure of nucleic add or amino acid sequences of 109P1D4 gene products are observed by the Northern, Southern, Western, PCR and DNA sequencing protocols discussed herein. In addition, other methods for observing perturbations in nudeotide and amino acid sequences such as single strand conformation polymorphism analysis are well known in the art (see, U.S. Patent Nos. 5,382,510 issued 7 September 1999, and 5,952,170 issued 17 January 1995).

Additionally, one can examine the methylation status of a 109P1D4 gene in a biological sample. Aberrant demethylation and/or hypermethylation of CpG islands in gene 5' regulatory regions frequently occurs in immortalized and transformed cells, and can result in altered expression of various genes. For example, promoter hypermethylation of the pi-class glutathione S-transferase (a protein expressed in normal prostate but not expressed in >90% of prostate carcinomas) appears to permanently silence transcription of this gene and is the most frequently detected genomic alteration in prostate carcinomas (De Marzo et al., Am. J. Pathol. 155(6): 1985-1992 (1999)). In addition, this alteration is present in at least of cases of high-grade prostatic intraepithelial neoplasia (PIN) (Brooks et al., Cancer Epidemlol. Biomarkers Prev., 1998, 7:531-536). In another example, expression of the LAGE-I tumor specific gene (which is not expressed in normal prostate but is expressed in 25-50% of prostate cancers) is induced by deoxy-azacytidine in lymphoblastoid cells, suggesting that tumoral expression is due to demethylation (Lethe et al., Int J. Cancer 76(6): 903-908 (1998)). A variety of assays for examining methylation status of a gene are wel known in the art For example, one can utilize, in Southern hybridization approaches, methylation-sensitive restriction enzymes that cannot cleave sequences that contain methylated CpG sites to assess the methylation status of CpG Islands. In addition, MSP (methylation specific PCR) can rapidly profile the methylation status of all the CpG sites present In a CpG island of a given gene. This procedure involves initial modification of DNA by sodium bisulfite (which will convert all unmelhylated cytosines to uracil) followed by amplification using primers specific for methylated versus unmethylated DNA. Protocols involving methylation interference can also be found for example in Current Protocols In Molecular Biology, Unit 12, Frederick M. Ausubel et al. eds., 1995.

Gene amplification is an additional method for assessing the status of 109P1D4. Gene amplification is measured in a sample directly, for example, by conventional Southern blotting or Northern blotting to quantitate the transcription of mRNA (Thomas, 1980, Proc. Natl. Acad. Sci. USA, 77:5201-5205), dot blotting (DNA analysis), or n situ hybridization, using an appropriately labeled probe, based on the sequences provided herein. Alternatively, antibodies are employed that recognize specific duplexes, including DNA duplexes, RNA duplexes, and DNA-RNA hybrid duplexes or DNA-protein

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00 duplexes. The antibodies in turn are labeled and the assay carried out where the duplex is bound to a surface, so that upon the formation of duplex on the surface, the presence of antibody bound to the duplex can be detected.

C Biopsied tissue or peripheral blood can be conveniently assayed for the presence of cancer cells using for example, Northern, dot blot or RT-PCR analysis to detect 109P1D4 expression. The presence of RT-PCR amplifiable 109P1D4 mRNA provides an Indication of the presence of cancer. RT-PCR assays are well known in the art RT-PCR detection assays for tumor cells in peripheral blood are currently being evaluated for use in the diagnosis and management of a number of human solid tumors. In the prostate cancer field, these include RT-PCR assays for the detection of cells expressing PSA and PSM (Verkaik et al., 1997, Urol. Res. 25:373-384; Ghossein et al., 1995, J. Clin. Oncol. 13:1195-2000; Heston et al., 1995, Clin. Chem. 41:1687- S1688).

0 A further aspect of the invention is an assessment of the susceptibility that an Individual has for developing cancer. In one embodiment a method for predicting susceptibility to cancer comprises detecting 109P1D4 mRNA or 109P1D4 protein in a CN tissue sample, its presence indicating susceptibility to cancer, wherein the degree of 109P1D4 mRNA expression correlates to the 00 0 degree of susceptibility. In a specific embodiment, the presence of 109P1 D4 n prostate or other tissue is examined, with the Spresence of 109P1 D4 in the sample providing an indication of prostate cancer susceptibility (or the emergence or existence of a prostate tumor). Similarly, one can evaluate the integrity 109P1D4 nucleotide and amino add sequences in a biological sample, in order to identify perturbations in the structure of these molecules such as Insertions, deletions, substitutions and the like. The presence of one or more perturbations In 109P1D4 gene products in the sample is an indication of cancer susceptibility (or the emergence or existence of a tumor).

The invention also comprises methods for gauging tumor aggressiveness. In one embodiment, a method for gauging aggressiveness of a tumor comprises determining the level of 109P1 D4 mRNA or 109P1D4 protein expressed by tumor cells, comparing the level so determined to the level of 109P1D4 mRNA or 109P1D4 protein expressed in a corresponding normal tissue taken from the same individual or a normal tissue reference sample, wherein the degree of 109P1D4 mRNA or 109P1 D4 protein expression in the tumor sample relative to the normal sample Indicates the degree of aggressiveness. In a specific embodiment, aggressiveness of a tumor is evaluated by determining the extent to which 109P1D4 is expressed in the tumor cells, with higher expression levels indicating more aggressive tumors. Another embodiment is the evaluation of the integrity of 109P1D4 nucleotide and amino add sequences in a biological sample, in order to identify perturbations in the structure of these molecules such as insertions, deletions, substitutions and the like. The presence of one or more perturbations indicates more aggressive tumors.

Another embodiment of the invention is directed to methods for observing the progression of a malignancy in an Individual over time. In one embodiment, methods for observing the progression of a malignancy in an Individual over time comprise determining the level of 109P1D4 mRNA or 109P1D4 protein expressed by cells in a sample of the tumor, comparing the level so determined to the level of 109P1D4 mRNA or 109P1D4 protein expressed in an equivalent tissue sample taken from the same individual at a different time, wherein the degree of 109P1D4 mRNA or 109P1D4 protein expression in the tumor sample over time provides information on the progression of the cancer. In a specific embodiment, the progression of a cancer is evaluated by determining 109P1 D4 expression in the tumor cells over time, where increased expression over time indicates a progression of the cancer. Also, one can evaluate the integrity 109P1D4 nucleotide and amino add sequences in a biological sample in order to identify perturbations in the structure of these molecules such as insertions, deletions, substitutions and the like, where the presence of one or more perturbations indicates a progression of the cancer.

The above diagnostic approaches can be combined with any one of a wide variety of prognostic and diagnostic protocols known In the art For example, another embodiment of the invention is directed to methods for observing a coincidence between the expression of 109P1D4 gene and 109P1D4 gene products (or perturbations in 109P1D4 gene and 109P1D4 gene 00 products) and a factor that is associated with malignancy, as a means for diagnosing and prognosticating the status of a tissue sample. A wide variety of factors associated with malignancy can be utilized, such as the expression of genes associated with Smalignancy PSA, PSCA and PSM expression for prostate cancer etc.) as well as gross cytological observations (see, e.g., S Bocking et al., 1984, Anal. Quant Cytol. 6(2):74-88; Epstein, 1995, Hum. Pathol. 26(2):223-9; Thorson et 1998, Mod.

CF) Pathol. 11(6):543-51; Balsden et al., 1999, Am. J. Surg. Pathol. 23(8):918-24). Methods for observing a coincidence between V. the expression of 109P1D4 gene and 109P1D4 gene products (or perturbations in 109P1D4 gene and 109P1D4 gene products) and another factor that is associated with malignancy are useful, for example, because the presence of a set of specific factors that coincide with disease provides information crucial for diagnosing and prognosticating the status of a tissue sample.

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In one embodiment, methods for observing a coincidence between the expression of 109P1D4 gene and 109P1 D4 0 gene products (or perturbations in 109P1 D4 gene and 109P1D4 gene products) and another factor associated with malignancy Sentails detecting the overexpression of 109P1D4 mRNA or protein in a tissue sample, detecting the overexpression of PSA mRNA C or protein in a tissue sample (or PSCA or PSM expression), and observing a coincidence of 109P1D4 mRNA or protein and PSA 00 mRNA or protein overexpression (or PSCA or PSM expression). In a specific embodiment the expression of 109P1D4 and PSA SmRNA In prostate tissue is examined, where the coincidence of 109P1D4 and PSA mRNA overexpression in the sample indicates the existence of prostate cancer, prostate cancer susceptibility or the emergence or status of a prostate tumor.

Methods for detecting and quantifying the expression of 109P1D4 mRNA or protein are described herein, and standard nucleic acid and protein detection and quantification technologies are well known in the art Standard methods for the detection and quantification of 109P1D4 mRNA include in situ hybridization using labeled 109P1D4 riboprobes, Northem blot and related techniques using 109P1 D4 poynucleotide probes, RT-PCR analysis using primers specific for 109P1D4, and other amplification type detection methods, such as, for example, branched DNA, SISBA, TMA and the like. In a specific embodiment, semiquantitative RT-PCR is used to detect and quantify 109P1 D4 mRNA expression. Any number of primers capable of amplifying 109P1D4 can be used for this purpose, including but not limited to the various primer sets specifically described herein. In a specific embodiment, polyclonal or monoclonal antibodies specifically reactive with the wild-type 109P1 D4 protein can be used in an immunohistochemical assay of biopsied tissue.

IX) Identification of Molecules That Interact With 109P1D4 The 109P1D4 protein and nucleic add sequences disclosed herein allow a skilled artisan to Identify proteins, small molecules and other agents that interact with 109P1D4, as well as pathways activated by 109P1 D4 via any one of a variety of art accepted protocols. For example, one can utilize one of the so-called interaction trap systems (also referred to as the "two-hybrid assay"). In such systems, molecules interact and reconstitute a transcription factor which directs expression of a reporter gene, whereupon the expression of the reporter gene is assayed. Other systems identify protein-protein Interactions in viv through reconstitution of a eukaryotic transcriptional activator, see, U.S. Patent Nos. 5,955,280 Issued 21 September 1999, 5,925,523 Issued 20 July 1999, 5,846,722 issued 8 December 1998 and 6,004,746 Issued 21 December 1999. Algorithms are also available In the art for genome-based predictions of protein function (see, Marcotte, et al., Nature 402:4 November 1999,83-86).

Alternatively one can screen peptide libraries to identify molecules that Interact with 109P1D4 protein sequences.

In such methods, peptides that bind to 109P1D4 are identified by screening libraries that encode a random or controlled collection of amino adds. Peptides encoded by the libraries are expressed as fusion proteins of bacteriophage coat proteins, the bacteriophage particles are then screened against the 109P1D4 protein(s).

Accordingly, peptides having a wide variety of uses, such as therapeutic, prognostic or diagnostic reagents, are thus identified without any prior Information on the structure of the expected ligand or receptor molecule. Typical peptide 00 libraries and screening methods that can be used to identify molecules that interact with 109P1D4 protein sequences are disclosed for example in U.S. Patent Nos. 5,723,286 issued 3 March 1998 and 5,733,731 issued 31 March 1998.

c Alternatively, cell lines that express 109P1 D4 are used to identify protein-protein interactions mediated by 109P1D4. Such interactions can be examined using immunoprecipitation techniques (see, Hamilton et al Blochem. Biophys. Res. Commun. 1999, 261:646-51). 109P1D4 protein can be immunoprecipitated from 109P1D4- V expressing cell lines using anti-109P1 D4 antibodies. Alternatively, antibodies against His-tag can be used in a cell line engineered to express fusions of 109P1D4 and a His-tag (vectors mentioned above). The immunoprecipitated complex can be examined for protein association by procedures such as Western blotting, 5 S-methionine labeling of proteins, protein Smicrosequencing, silver staining and two-dimensional gel electrophoresis.

SSmall molecules and ligands that interact with 109P1D4 can be identified through related embodiments of such screening assays. For example, small molecules can be identified that interfere with protein function, including molecules

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that interfere with 109P1D4's ability to mediate phosphorylation and de-phosphorylation, interaction with DNA or RNA 00 0 molecules as an indication of regulation of cell cycles, second messenger signaling or tumorigenesis. Similarly, small Smolecules that modulate 109P1 D4-related ion channel, protein pump, or cell communication functions are identified and used to treat patients that have a cancer that expresses 109P1D4 (see, Hille, Ionic Channels of Excitable Membranes 2 nd Ed., Sinauer Assoc., Sunderand, MA, 1992). Moreover, ligands that regulate 109P1D4 function can be identified based on their ability to bind 109P1D4 and activate a reporter construct Typical methods are discussed for example in U.S. Patent No. 5,928,868 issued 27 July 1999, and include methods for forming hybrid Ilgands in which at least one ligand is a small molecule. In an illustrative embodiment, cells engineered to express a fusion protein of 109P1D4 and a DNA-binding protein are used to co-express a fusion protein of a hybrid ligand/small molecule and a cDNA library transcriptional activator protein. The cells further contain a reporter gene, the expression of which is conditioned on the proximity of the first and second fusion proteins to each other, an event that occurs only if the hybrid ligand binds to target sites on both hybrid proteins. Those cells that express the reporter gene are selected and the unknown small molecule or the unknown ligand is identified. This method provides a means of identifying modulators, which activate or Inhibit 109P1D4.

An embodiment of this invention comprises a method of screening for a molecule that interacts with a 109P1 D4 amino acid sequence shown in Figure 2 or Figure 3, comprising the steps of contacting a population of molecules with a 109P1 D4 amino acid sequence, allowing the population of molecules and the 109P1D4 amino acid sequence to interact under conditions that facilitate an interaction, determining the presence of a molecule that interacts with the 109P104 amino add sequence, and then separating molecules that do not Interact with the 109P1D4 amino add sequence from molecules that do. In a specific embodiment, the method further comprises purifying, characterizing and identifying a molecule that interacts with the 109P1D4 amino acid sequence. The identified molecule can be used to modulate a function performed by 109P1D4. In a preferred embodiment, the 109P1D4 amino acid sequence is contacted with a library of peptides.

2) Therapeutic Methods and Compositions The Identification of 109P1D4 as a protein that is normally expressed in a restricted set of tissues, but which is also expressed in cancers such as those listed in Table I, opens a number of therapeutic approaches to the treatment of such cancers.

Of note, targeted antitumor therapies have been useful even when the targeted protein Is expressed on normal tissues, even vital normal organ tissues. A vital organ is one that is necessary to sustain life, such as the heart or colon. A non-vital organ Is one that can be removed whereupon the individual is still able to survive. Examples of non-vital organs are ovary, breast, and prostate.

49 00 For example, Herceptin® is an FDA approved pharmaceutical that has as its active ingredient an antibody which is immunoreactive with the protein variously known as HER2, HER2/neu, and erb-b-2. It is marketed by Genentech and has 1 been a commercially successful antitumor agent Herceptin sales reached almost $400 million in 2002. Herceptin is a u treatment for HER2 positive metastatic breast cancer. However, the expression of HER2 is not limited to such tumors. The C)same protein is expressed In a number of normal tissues. In particular, it is known that HER2/neu is present in normal in kidney and heart, thus these tissues are present in all human recipients of Herceptin. The presence of HER2/neu in normal kidney is also confirmed by Latif, et al., B.J.U. International (2002) 89:5-9. As shown in this article (which evaluated whether renal cell carcinoma should be a preferred indication for anti-HER2 antibodies such as Herceptin) both protein and SmRNA are produced in benign renal tissues. Notably, HER2/neu protein was strongly overexpressed in benign renal tissue.

SDespite the fact that HER2/neu is expressed in such vital tissues as heart and kidney, Herceptin is a very useful, FDA approved, and commercially successful drug. The effect of Herceptin on cardiac tissue, "cardiotoxidty," has merely

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1 been a side effect to treatment When patients were treated with Herceptin alone, significant cardiotoxicity occurred in a very 00 0low percentage of patients.

0 Of particular note, although kidney tissue is indicated to exhibit normal expression, possibly even higher expression than cardiac tissue, kidney has no appreciable Herceptin side effect whatsoever. Moreover, of the diverse array of normal tissues in which HER2 is expressed, there is very little occurrence of any side effect Only cardiac tissue has manifested any appreciable side effect at all. A tissue such as kidney, where HER2/neu expression is especially notable, has not been the basis for any side effect Furthermore, favorable therapeutic effects have been found for antitumor therapies that target epidermal growth factor receptor (EGFR). EGFR is also expressed in numerous normal tissues. There have been very fimited side effects in normal tissues following use of anti-EGFR therapeutics.

Thus, expression of a target protein in normal tissue, even vital normal tissue, does not defeat the utility of a targeting agent for the protein as a therapeutic for certain tumors in which the protein is also overexpressed.

Accordingly, therapeutic approaches that inhibit the activity of a 109P1D4 protein are useful for patients suffering from a cancer that expresses 109P1D4. These therapeutic approaches generally fall into two classes. One class comprises various methods for inhibiting the binding or association of a 109P1D4 protein with its binding partner or with other proteins.

Another class comprises a variety of methods for inhibiting the transcription of a 109P1D4 gene or translation of 109P1 D4 mRNA.

Anti-Cancer Vaccines The invention provides cancer vaccines comprising a 109P D4-related protein or 109P1D4-related nucleic add. In view of the expression of 109P1D4, cancer vaccines prevent and/or treat 109P1D4-expressing cancers with minimal or no effects on non-target tissues. The use of a tumor antigen in a vaccine that generates humoral andlor cell-mediated immune responses as anti-cancer therapy is well known in the art and has been employed in prostate cancer using human PSMA and rodent PAP immunogens (Hodge et al., 1995, Int J. Cancer 63:231-237; Fong et 1997, J. Immunol. 159:3113-3117).

Such methods can be readily practiced by employing a 109P1D4-related protein, or a 109P1D4-encoding nucleic acid molecule and recombinant vectors capable of expressing and presenting the 109P1 D4 Immunogen (which typically comprises a number of antibody or T cell epitopes). Skilled artisans understand that a wide variety of vaccine systems for delivery of immunoreactive epitopes are known in the art (see, Heryln et al., Ann Med 1999 Feb 31(1):66-78; Maruyama et al., Cancer Immunol Immunother 2000 Jun 49(3):123-32) Briefly, such methods of generating an Immune response (e.g.

humoral andlor cell-mediated) in a mammal, comprise the steps of: exposing the mammal's immune system to an 00 0 immunoreactive epitope an epitope present in a 109P1D4 protein shown in Figure 3 or analog or homolog thereof) so Sthat the mammal generates an Immune response that is specific for that epitope generates antibodies that specifically recognize that epitope). In a preferred method, a 109P1D4 immunogen contains a biological motif, see Tables VIII-XXI C) and XXII-XUX, or a peptide of a size range from 109P1 D4 Indicated In Figure 5, Figure 6, Figure 7, Figure 8, and Figure 9.

The entire 109P1 D4 protein, immunogenic regions or epitopes thereof can be combined and delivered by various means. Such vaccine compositions can include, for example, lipopeptides (e.g.,Vitiello, A. et al., J. Clin. Invest. 95:341, 1995), peptide compositions encapsulated In poly(DL-lactide-co-glycolide) ("PLG) microspheres (see, Eldridge, et al., Molec. Immunol. 28:287-294,1991: Alonso et al., Vaccine 12:299-306, 1994; Jones et Vaccine 13:675-681, 1995), CKl peptide compositions contained in Immune stimulating complexes (ISCOMS) (see, Takahashi et Nature 344:873- C 875, 1990; Hu et al., Clin Exp Immunol. 113:235-243, 1998), multiple antigen peptide systems (MAPs) (see Tam, J. P., Proc. Natl. Acad. Sci. U.S.A. 85:5409-5413, 1988; Tam, J. Immunol. Methods 196:17-32,1996), peptides formulated as 00 multivalent peptides; peptides for use in ballistic delivery systems, typically crystallized peptides, viral delivery vectors S(Perkus, M. E. et al., In: Concepts in vaccine development, Kaufmann, S. H. ed., p. 379, 1996; Chakrabarti, S. et al., CLN Nature 320:535, 1986; Hu, S. L et al., Nature 320:537, 1986; Kieny, et al., AIDS Bio/echnology 4:790, 1986; Top, F.

H. et J. Infect. Dis. 124:148, 1971; Chanda, P. K. et al., Virology 175:535, 1990), particles of viral or synthetic origin Kofler, N. et J. Immunol. Methods. 192:25, 1996; Eldridge, J. H. etal., Sem. Hematol. 30:16, 1993; Falo, L. Jr. et al., Nature Med. 7:649, 1995), adjuvants (Warren, H. Vogel, F. and Chedid, L A. Annu. Rev. Immunol. 4:369,1986; Gupta, R. K. et al., Vaccine 11:293,1993), liposomes (Reddy, R. et al., J. Immunol. 148:1585, 1992; Rock, K. Immunol.

Today 17:131, 1996), or, naked or particle absorbed cDNA (Ulmer, J. B. et al., Science 259:1745, 1993; Robinson, H. L., Hunt, L and Webster, R. Vaccine 11:957, 1993; Shiver, J. W. et al., In: Concepts in vaccine development, Kaufmann, S. H. ed., p. 423, 1996; Cease, K. and Berzofsky, J. Annu. Rev. Immunol. 12:923, 1994 and Eldridge, J. H. et a., Sem. Hematol. 30:16, 1993). Toxin-targeted delivery technologies, also known as receptor mediated targeting, such as those of Avant Immunotherapeutics, Inc. (Needham, Massachusetts) may also be used.

In patients with 109P1D4-associated cancer, the vaccine compositions of the Invention can also be used in conjunction with other treatments used for cancer, surgery, chemotherapy, drug therapies, radiation therapies, etc.

including use In combination with immune adjuvants such as IL-2, IL-12, GM-CSF, and the like.

Cellular Vaccines: CTL epitopes can be determined using specific algorithms to identify peptides within 109P1D4 protein that bind corresponding HLA alleles (see Table IV; Epimer T and Epimatri, Brown University (URL brown.edu/Research/TB- HIV_Lablepimatrixepimatrixhtml); and, BIMAS, (URL bimas.dcrtnih.govl; SYFPEITHI at URL syfpeithi.bmi-heldelberg.com/).

In a preferred embodiment, a 109P1D4 immunogen contains one or more amino add sequences identified using techniques well known in the art, such as the sequences shown in Tables VIII-XXI and XXII-XUX or a peptide of 8, 9, 10 or 11 amino acids spedfied by an HLA Class I motif/supermotif Table IV Table IV or Table IV and/or a peptide of at least 9 amino adds that comprises an HLA Class II motif/supermotif Table IV or Table IV As is appreciated in the art, the HLA Class I binding groove is essentially closed ended so that peptides of only a particular size range can fit Into the groove and be bound, generally HLA Class I epitopes are 8, 9, 10, or 11 amino acids long. In contrast, the HLA Class II binding groove is essentially open ended; therefore a peptide of about 9 or more amino acids can be bound by an HLA Class II molecule. Due to the binding groove differences between HLA Class I and II, HLA Class I motifs are length spedfic, i.e., position two of a Class I motif is the second amino acid in an amino to carboxyl direction of the peptide. The amino add positions in a Class II motif are relative only to each other, not the overall peptide, additional amino adds can be attached to the amino and/or carboxyl termini of a motif-bearing sequence. HLA Class II epitopes are often 9,10, 11,12,13, 00 14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids long, or longer than 25 amino acids.

O Antibody-based Vaccines C- A wide variety of methods for generating an immune response in a mammal are known in the art (for example as the first step in the generation of hybridomas). Methods of generating an immune response in a mammal comprise exposing the mammal's immune system to an immunogenic epitope on a protein a 109P1D4 protein) so that an immune n response is generated. A typical embodiment consists of a method for generating an immune response to 109P1D4 in a Shost, by contacting the host with a sufficient amount of at least one 109P1D4 B cell or cytotoxic T-ceU epitope or analog thereof; and at least one periodic interval thereafter re-contacting the host with the 109P1D4 B cell or cytotoxic T-cell epitope Sor analog thereof. A specific embodiment consists of a method of generating an immune response against a 109P1D4related protein or a man-made multiepitopic peptide comprising: administering 109P1 D4 immunogen a 109P1 D4 cK protein or a peptide fragment thereof, a 109P1D4 fusion protein or analog etc.) in a vaccine preparation to a human or another mammal. Typically, such vaccine preparations further contain a suitable adjuvant (see, U.S. Patent No.

00 6,146,635) or a universal helper epitope such as a PADREm peptide (Epimmune Inc., San Diego, CA; see, Alexander Set al., J. Immunol. 2000 164(3); 164(3): 1625-1633; Alexander et al., Immunity 1994 751-761 and Alexander et al., Immunol. Res. 1998 18(2): 79-92). An alternative method comprises generating an immune response in an individual against a 109P1D4 immunogen by: administering in vivo to muscle or skin of the individuals body a DNA molecule that comprises a DNA sequence that encodes a 109P1D4 immunogen, the DNA sequence operatively linked to regulatory sequences which control the expression of the DNA sequence; wherein the DNA molecule is taken up by cells, the DNA sequence is expressed in the cells and an immune response is generated against the immunogen (see, U.S. Patent No.

5,962,428). Optionally a genetic vaccine facilitator such as anionic lipids; saponins; lectins; estrogenic compounds; hydroxylated lower alkyls; dimethyl sulfoxide; and urea is also administered. In addition, an antiidlotypic antibody can be administered that mimics 109P1D4, in order to generate a response to the target antigen.

Nucleic Acid Vaccines: Vaccine compositions of the invention include nucleic acid-mediated modalities. DNA or RNA that encode protein(s) of the invention can be administered to a patient Genetic immunization methods can be employed to generate prophylactic or therapeutic humoral and cellular immune responses directed against cancer cells expressing 109P1D4.

Constructs comprising DNA encoding a 109P1D4-related protein/immunogen and appropriate regulatory sequences can be Injected directly into muscle or skin of an individual, such that the cells of the muscle or skin take-up the construct and express the encoded 109P1D4 protein/Immunogen. Alternatively, a vaccine comprises a 109P1D4-related protein.

Expression of the 109P1D4-related protein immunogen results In the generation of prophylactic or therapeutic humoral and cellular immunity against cells that bear a 109P1D4 protein. Various prophylactic and therapeutic genetic immunization techniques known in the art can be used (for review, see information and references published at Intemet address genweb.com). Nucleic acid-based delivery is described, for instance, in Wolff et. al., Science 247:1465 (1990) as well as U.S. Patent Nos. 5,580,859; 5,589,466; 5,804,566; 5,739,118; 5,736,524; 5,679,647; WO 98/04720. Examples of DNAbased delivery technologies include naked DNA', facilitated (buplvicaine, polymers, peptide-mediated) delivery, cationic lipid complexes, and particle-mediated ('gene gun) or pressure-mediated delivery (see, U.S. Patent No. 5,922,687).

For therapeutic or prophylactic Immunization purposes, proteins of the invention can be expressed via viral or bacterial vectors. Various viral gene delivery systems that can be used in the practice of the invention include, but are not limited to, vaccinia, fowlpox, canarypox, adenovirus, influenza, poliovirus, adeno-assodated virus, lentivirus, and sindbis virus (see, e.g., Restifo, 1996, Curr. Opin. Immuno. 8:658-663; Tsang et al. J, Nall. Cancer Inst 87:982-990 (1995)). Non-viral delivery systems can also be employed by introducing naked DNA encoding a 109P1D4-related protein Into the patient intramuscularly or 00 intradermally) to induce an anti-tumor response.

SVaccinia virus is used, for example, as a vector to express nuceotide sequences that encode the peptides of the C- invention. Upon introduction into a host the recombinant vaccinia virus expresses the protein Immunogenic peptide, and thereby elicits a host immune response. Vacdnia vectors and methods useful in immunization protocols are described in, U.S. Patent No. 4,722,848. Another vector is BCG (Bacille Calmette Guerin). BCG vectors are described in Stover et al., Nature 351:456-460 (1991). A wide variety of other vectors useful for therapeutic administration or immunization of the peptides of the invention, e.g. adeno and adeno-assodated virus vectors, retroviral vectors, Salmonella typhi vectors, detoxified anthrax toxin vectors, and the like, will be apparent to those skilled in the art from the description herein.

SThus, gene delivery systems are used to deliver a 109P1D4-related nucleic acid molecule. In one embodiment the full- Slength human 109P104 cDNA is employed. In another embodiment, 109P1D4 nucleic acid molecules encoding specific cytotoxic T lymphocyte (CTL) andlor antibody epitopes are employed.

NC ExVivo Vaccines 0 Various ex vio strategies can also be employed to generate an immune response. One approach Involves the use of Santigen presenting cells (APCs) such as dendritic cells (DC) to present 109P1D4 antigen to a patient's Immune system. Dendritic cells express MHC class I and II molecules, B7 co-stimulator, and IL-12 and are thus highly specialized antigen presenting cells.

In prostate cancer, autologous dendritic cells pulsed with peptides of the prostate-specific membrane antigen (PSMA) are being used in a Phase I clinical trial to stimulate prostate cancer patients' immune systems (Tjoa et al., 1996, Prostate 28:65- 69; Murphy et al., 1996, Prostate 29:371-380). Thus, dendritic cells can be used to present 109P1D4 peptides to T cells in the context of MHC class I or II molecules. In one embodiment autologous dendritic cells are pulsed with 109P1 D4 peptides capable of binding to MHC class I and/or class II molecules. In another embodiment dendritic cells are pulsed with the complete 109P1D4 protein. Yet another embodiment involves engineering the overexpression of a 109P1D4 gene in dendritic cells using various implementing vectors known in the art, such as adenovrug (Arthur et al., 1997, Cancer Gene Ther. 4:17-25), retrovirus (Henderson et al., 1996, Cancer Res. 56:3763-3770), lentivirus, adeno-assodated virus, DNA transfection (Ribas et 1997, Cancer Res. 57:2865-2869), or tumor-derived RNA transfection (Ashley et 1997, J. Exp.

Med. 186:1177-1182). Cells that express 109P1D4 can also be engineered to express immune modulators, such as GM- CSF, and used as immunizing agents.

XB.) 109P1 D4 as a Tarnet for Antibodv-based Therapy 109P1 D4 Is an attractive target for antibody-based therapeutic strategies. A number of antibody strategies are known in the art for targeting both extracellular and intracellular molecules (see, complement and ADCC mediated killing as well as the use of intrabodies). Because 109P1D4 is expressed by cancer cells of various lineages relative to corresponding normal cells, systemic administration of 109P D4-immunoreactive compositions are prepared that exhibit excellent sensitivity without toxic, non-specific and/or non-target effects caused by binding of the immunoreactive composition to non-target organs and tissues. Antibodies specifically reactive with domains of 109P1D4 are useful to treat 109P1D4-expressing cancers systemically, either as conjugates with a toxin or therapeutic agent, or as naked antibodies capable of inhibiting cell proliferation or function.

109P1D4 antibodies can be introduced into a patient such that the antibody binds to 109P1D4 and modulates a function, such as an interaction with a binding partner, and consequently mediates destruction of the tumor cells and/or inhibits the growth of the tumor cells. Mechanisms by which such antibodies exert a therapeutic effect can include complement-mediated cytolysis, antibody-dependent cellular cytotoxicity, modulation of the physiological function of 109P1D4, Inhibition of ligand binding or signal transduction pathways, modulation of tumor cell differentiation, alteration of 00 tumor angiogenesis factor profiles, and/or apoptosis.

Those skilled in the art understand that antibodies can be used to specifically target and bind immunogenic c molecules such as an immunogenic region of a 109P1D4 sequence shown in Figure 2 or Figure 3. In addition, skilled artisans understand that it is routine to conjugate antibodies to cytotoxic agents (see, Slevers et al. Blood 93:11 3678- 3684 (June 1, 1999)). When cytotoxic and/or therapeutic agents are delivered directly to cells, such as by conjugating them V' to antibodies specific for a molecule expressed by that cell 109P1 D4), the cytooxic agent will exert its known biological effect cytotoxicity) on those cells.

A wide variety of compositions and methods for using antibody-cytotoxic agent conjugates to kill cells are known in Sthe art. In the context of cancers, typical methods entail administering to an animal having a tumor a biologically effective Samount of a conjugate comprising a selected cytotoxic and/or therapeutic agent linked to a targeting agent an anti- S109P1 D4 antibody) that binds to a marker 109P1D4) expressed, accessible to binding or localized on the cell surfaces.

SA typical embodiment is a method of delivering a cytotoxic and/or therapeutic agent to a cell expressing 109P1D4, Scomprising conjugating the cytotoxic agent to an antibody that immunospecifically binds to a 109P1 D4 epitope, and, Sexposing the cell to the antibody-agent conjugate. Another illustrative embodiment is a method of treating an individual suspected of suffering from metastasized cancer, comprising a step of administering parenterally to said individual a pharmaceutical composition comprising a therapeutically effective amount of an antibody conjugated to a cytotoxic and/or therapeutic agent Cancer immunotherapy using anti-109P1D4 antibodies can be done in accordance with various approaches that have been successfully employed in the treatment of other types of cancer, including but not limited to colon cancer (Arlen et al., 1998, Crit Rev. Immunol. 18:133-138), multiple myeloma (Ozaki at 1997, Blood 90:3179-3186, Tsunenari et a., 1997, Blood 90:2437-2444), gastric cancer (Kasprzyk et al., 1992, Cancer Res. 52:2771-2776), B-cell lymphoma (Funakoshi eta/., 1996, J. Immunother. Emphasis Tumor Immunol. 19:93-101), leukemia (Zhong eta/., 1996, Leuk. Res. 20:581-589), colorectal cancer (Moun et al., 1994, Cancer Res. 54:6160-6166; Velders et al., 1995, Cancer Res. 55:4398-4403), and breast cancer (Shepard et al., 1991, J. Clin. Immunol. 11:117-127). Some therapeutic approaches involve conjugation of naked antibody to a toxin or radioisotope, such as the conjugation of Y91 or 113 to anti-CD20 antibodies ZevalinTM, IDEC Pharmaceuticals Corp. or BexxarTu, Coulter Pharmaceuticals), while others involve co-administration of antibodies and other therapeutic agents, such as Herceptinm (trastuzumab) with paditaxel (Genentech, Inc.). The antibodies can be conjugated to a therapeutic agent To treat prostate cancer, for example, 109P1D4 antibodies can be administered in conjunction with radiation, chemotherapy or hormone ablation. Also, antibodies can be conjugated to a toxin such as calicheamidn Mylotargm, Wyeth-Ayerst, Madison, NJ, a recombinant humanized lgG4 kappa antibody conjugated to antitumor antibiotic calicheamidn) or a maytansinold taxane-based Tumor-Activated Prodrug, TAP, platform, ImmunoGen, Cambridge, MA, also see US Patent 5,416,064).

Although 109P1D4 antibody therapy is useful for all stages of cancer, antibody therapy can be particularly appropriate in advanced or metastatic cancers. Treatment with the antibody therapy of the Invention is indicated for patients who have received one or more rounds of chemotherapy. Alternatively, antibody therapy of the invention is combined with a chemotherapeutic or radiation regimen for patients who have not received chemotherapeutic treatment Additionally, antibody therapy can enable the use of reduced dosages of concomitant chemotherapy, particularly for patients who do not tolerate the toxicity of the chemotherapeutic agent very well. Fan et al. (Cancer Res. 53:4637-4642, 1993), Prewett et al.

(Intemational J. of Onco. 9:217-224, 1996), and Hancock et al. (Cancer Res. 51:45754580,1991) describe the use of various antibodies together with chemotherapeutic agents.

Although 109P1 D4 antibody therapy is useful for all stages of cancer, antibody therapy can be particularly )4 OO appropriate in advanced or metastatic cancers. Treatment with the antibody therapy of the invention is indicated for patients O who have received one or more rounds of chemotherapy. Alternatively, antibody therapy of the invention is combined with a 1 chemotherapeutic or radiation regimen for patients who have not received chemotherapeutic treatment Additionally, antibody therapy can enable the use of reduced dosages of concomitant chemotherapy, particularly for patients who do not y) tolerate the toxicity of the chemotherapeutic agent very well.

Cancer patients can be evaluated for the presence and level of 109P1D4 expression, preferably using immunohistochemical assessments of tumor tissue, quantitative 109P1D4 imaging, or other techniques that reliably indicate the presence and degree of 109P1D4 expression. Immunohistochemical analysis of tumor biopsies or surgical specimens is 0 preferred for this purpose. Methods for Immunohistochemical analysis of tumor tissues are well known in the art.

Anti-109P1D4 monoclonal antibodies that treat prostate and other cancers include those that initiate a potent immune response against the tumor or those that are directly cytotoxic. In this regard, anti-109P1D4 monoclonal antibodies 1 (mAbs) can elicit tumor cell lysis by either complement-mediated or antibody-dependent cell cytotoxicity (ADCC) 00 Smechanisms, both of which require an intact Fc portion of the immunoglobulin molecule for interaction with effector cell Fc Sreceptor sites on complement proteins. In addition, anti-109P1D4 mAbs that exert a direct biological effect on tumor growth are useful to treat cancers that express 109P1 D4. Mechanisms by which directly cytotoxic mAbs act include: inhibition of cell growth, modulation of cellular differentiation, modulation of tumor angiogenesis factor profiles, and the induction of apoptosis.

The mechanism(s) by which a particular anti-109P1D4 mAb exerts an anti-tumor effect is evaluated using any number of in vitm assays that evaluate cell death such as ADCC, ADMMC, complement-mediated cell lysis, and so forth, as Is generally known in the art In some patients, the use of murine or other non-human monoclonal antibodies, or human/mouse chimeric mAbs can induce moderate to strong immune responses against the non-human antibody. This can result In clearance of the antibody from circulation and reduced efficacy. In the most severe cases, such an immune response can lead to the extensive formation of immune complexes which, potentially, can cause renal failure. Accordingly, preferred monoclonal antibodies used in the therapeutic methods of the invention are those that are either fully human or humanized and that bind specifically to the target 109P1D4 antigen with high affinity but exhibit low or no antigenicity In the patient Therapeutic methods of the invention contemplate the administration of single anti-109P1 D4 mAbs as well as combinations, or cocktails, of different mAbs. Such mAb cocktails can have certain advantages inasmuch as they contain mAbs that target different epitopes, exploit different effector mechanisms or combine directly cytotoxic mAbs with mAbs that rely on immune effector functionality. Such mAbs in combination can exhibit synergistic therapeutic effects. In addition, anti- 109P1D4 mAbs can be administered concomitantly with other therapeutic modalities, including but not limited to various chemotherapeutic agents, androgen-blockers, immune modulators IL-2, GM-CSF), surgery or radiation. The anti- 109P1D4 mAbs are administered in their "naked" or unconjugated form, or can have a therapeutic agent(s) conjugated to them.

Anti- 09P1D4 antibody formulations are administered via any route capable of delivering the antibodies to a tumor cell. Routes of administration include, but are not limited to, intravenous, intraperitoneal, intramuscular, intratumor, Intradermal, and the like. Treatment generally involves repeated administration of the anti-109P1D4 antibody preparation, via an acceptable route of administration such as intravenous injection typically at a dose in the range of about 0.1, .2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or 25 mg/kg body weight In general, doses in the range of 10-1000 mg mAb per week are effective and well tolerated.

Based on clinical experience with the Herceptin

T

mAb in the treatment of metastatic breast cancer, an initial loading dose of approximately 4 mg/kg patient body weight IV, followed by weekly doses of about 2 mg/kg IV of the anti- 00 109P1D4 mAb preparation represents an acceptable dosing regimen. Preferably, the initial loading dose is administered as a 90-minute or longer infusion. The periodic maintenance dose is administered as a 30 minute or longer infusion, provided the initial dose was well tolerated. As appreciated by those of skill in the art, various factors can influence the ideal dose regimen in a particular case. Such factors include, for example, the binding affinity and half life of the Ab or mAbs used, the S degree of 109P1D4 expression in the patient, the extent of circulating shed 109P1D4 antigen, the desired steady-state m' antibody concentration level, frequency of treatment, and the influence of chemotherapeutic or other agents used in combination with the treatment method of the invention, as well as the health status of a particular patient Optionally, patients should be evaluated for the levels of 109P1D4 in a given sample the levels of circulating 109P1D4 antigen and/or 109P1D4 expressing cells) in order to assist in the determination of the most effective dosing 0 regimen, etc. Such evaluations are also used for monitoring purposes throughout therapy, and are useful to gauge therapeutic success in combination with the evaluation of other parameters (for example, urine cytology and/or ImmunoCyt C levels in bladder cancer therapy, or by analogy, serum PSA levels in prostate cancer therapy).

00 O Anti-idiotypic anti- 09P1 D4 antibodies can also be used in anti-cancer therapy as a vaccine for inducing an O immune response to cells expressing a 109P1D4-related protein. In particular, the generation of anti-idiotypic antibodies is well known in the art; this methodology can readily be adapted to generate anti-idiotypic anti-109P1D4 antibodies that mimic an epitope on a 109P1D4-related protein (see, for example, Wagner et at., 1997, Hybridoma 16: 33-40; Foon et al., 1995, J.

Clin. Invest 96:334-342; Herlyn et al., 1996, Cancer Immunol. Immunother. 43:65-76). Such an anti-Idiotypic antibody can be used in cancer vaccine strategies.

109P1D4 as a Target for Cellular Immune Responses Vaccines and methods of preparing vaccines that contain an immunogenically effective amount of one or more HLA-binding peptides as described herein are further embodiments of the invention. Furthermore, vaccines in accordance with the invention encompass compositions of one or more of the claimed peptides. A peptide can be present in a vaccine individually. Alternatively, the peptide can exist as a homopolymer comprising multiple copies of the same peptide, or as a heteropolymer of various peptides. Polymers have the advantage of increased immunological reaction and, where different peptide epitopes are used to make up the polymer, the additional ability to induce antibodies and/or CTLs that react with different antigenic determinants of the pathogenic organism or tumor-related peptide targeted for an immune response. The composition can be a naturally occurring region of an antigen or can be prepared, recombinantly or by chemical synthesis.

Carriers that can be used with vaccines of the Invention are well known in the art, and include, thyroglobulin, albumins such as human serum albumin, tetanus toxoid, polyamino acds such as poly L-fyslne, poly L-glutamic add, influenza, hepatitis B virus core protein, and the like. The vaccines can contain a physiologically tolerable acceptable) diluent such as water, or saline, preferably phosphate buffered saline. The vaccines also typically include an adjuvant Adjuvants such as incomplete Freund's adjuvant aluminum phosphate, aluminum hydroxide, or alum are examples of materials well known in the art. Additionally, as disclosed herein, CTL responses can be primed by conjugating peptides of the invention to lipids, such as tripalmitoyl-S-glyceryfcysteinlyseryl- serine (P3CSS). Moreover, an adjuvant such as a synthetic cytosine-phosphorothiolated-guanine-containing (CpG) oligonudeotldes has been found to increase CTL responses 10- to 100-fold. (see, e.g. Davila and Cells, J. Immunol. 165:539-547 (2000)) Upon immunization with a peptide composition in accordance with the Invention, via Injection, aerosol, oral, transdermal, transmucosal, intrapleural, intrathecal, or other suitable routes, the immune system of the host responds to the vaccine by producing large amounts of CTLs and/or HTLs specific for the desired antigen. Consequently, the host becomes 00 at least partially immune to later development of cells that express or overexpress 109P1D4 antigen, or derives at least 0some therapeutic benefit when the antigen was tumor-associated.

In some embodiments, it may be desirable to combine the class I peptide components with components that SInduce or facilitate neutralizing antibody and or helper T cell responses directed to the target antigen. A preferred Sembodiment of such a composition comprises class I and class II epitopes in accordance with the invention. An alternative Sembodiment of such a composition comprises a class I and/or class II epitope in accordance with the invention, along with a cross reactive HTL epitope such as PADRE T (Epimmune, San Diego, CA) molecule (described in U.S. Patent Number 5,736,142).

SA vaccine of the invention can also include antigen-presenting cells (APC), such as dendritic cells as a vehicle to present peptides of the invention. Vaccine compositions can be created in vitro, following dendritic cell mobilization and harvesting, whereby loading of dendritic cells occurs in vitro. For example, dendritic cells are transfected, 00 with a minigene in accordance with the invention, or are pulsed with peptides. The dendritic cell can then be Sadministered to a patient to elicit immune responses in vivo. Vaccine compositions, either DNA- or peptide-based, can also be administered in vivo in combination with dendritic cell mobilization whereby loading of dendritic cells occurs in vivo.

Preferably, the following principles are utilized when selecting an array of epitopes for inclusion in a polyepitopic composition for use in a vaccine, or for selecting discrete epitopes to be included in a vaccine and/or to be encoded by nucleic acids such as a minigene. It is preferred that each of the following principles be balanced in order to make the selection. The multiple epitopes to be incorporated In a given vaccine composition may be, but need not be, contiguous in sequence in the native antigen from which the epitopes are derived.

Epitopes are selected which, upon administration, mimic immune responses that have been observed to be correlated with tumor cearance. For HLA Class I this includes 3-4 epitopes that come from at least one tumor associated antigen (TAA). For HLA Class II a similar rationale is employed; again 3-4 epitopes are selected from at least one TAA (see, Rosenberg et Science 278:1447-1450). Epitopes from one TAA may be used in combination with epitopes from one or more additional TAAs to produce a vaccine that targets tumors with varying expression patterns of frequently-expressed TAAs.

Epitopes are selected that have the requisite binding affinity established to be correlated with immunogenicity: for HLA Class I an ICso of 500 nM or less, often 200 nM or less; and for Class II an ICso of 1000 nM or less.

Suffident supermotif bearing-peptides, or a sufficient array of allele-specific motif-bearing peptides, are selected to give broad population coverage. For example, it is preferable to have at least 80% population coverage. A Monte Carlo analysis, a statistical evaluation known in the art, can be employed to assess the breadth, or redundancy of, population coverage.

When selecting epitopes from cancer-related antigens it is often useful to select analogs because the patient may have developed tolerance to the native epitope.

Of particular relevance are epitopes referred to as 'nested epitopes." Nested epitopes occur where at least two epitopes overlap In a given peptide sequence. A nested peptide sequence can comprise B cell, HLA class I and/or HLA class II epitopes. When providing nested epitopes, a general objective Is to provide the greatest number of epitopes per sequence. Thus, an aspect is to avoid providing a peptide that is any longer than the amino terminus of the amino terminal epitope and the carboxyl terminus of the carboxyl terminal epitope in the peptide. When providing a multi-epitopic sequence, such as a sequence comprising nested epitopes, It is generally important to screen the sequence In order to insure that It does not have pathological or other deleterious biological properties.

If a polyepitopic protein is created, or when creating a minigene, an objective is to generate the smallest 00 peptide that encompasses the epitopes of interest. This principle is similar, if not the same as that employed when selecting Sa peptide comprising nested epitopes. However, with an artificial polyepitopic peptide, the size minimization objective is balanced against the need to integrate any spacer sequences between epitopes in the polyepitopic protein. Spacer amino C acid residues can, for example, be introduced to avoid junctional epitopes (an epitope recognized by the immune system, not Spresent in the target antigen, and only created by the man-made juxtaposition of epitopes), or to facilitate cleavage between l epitopes and thereby enhance epitope presentation. Junctional epitopes are generally to be avoided because the recipient Omay generate an immune response to that non-native epitope. Of particular concern is a junctional epitope that is a 'dominant epitope." A dominant epitope may lead to such a zealous response that immune responses to other epitopes are Sdiminished or suppressed.

Where the sequences of multiple variants of the same target protein are present, potential peptide cr epitopes can also be selected on the basis of their conservancy. For example, a criterion for conservancy may define that C- the entire sequence of an HLA class I binding peptide or the entire 9-mer core of a class II binding peptide be conserved in a 00 designated percentage of the sequences evaluated for a specific protein antigen.

0 XC.1. Minigene Vacdnes A number of different approaches are available which allow simultaneous delivery of multiple epitopes. Nudeic acids encoding the peptides of the invention are a particularly useful embodiment of the Invention. Epitopes for inclusion in a minigene are preferably selected according to the guidelines set forth in the previous section. A preferred means of administering nudeic acids encoding the peptides of the invention uses minigene constructs encoding a peptide comprising one or multiple epitopes of the invention.

The use of multi-epitope minigenes is described below and in, Ishioka et al., J. Immunol. 162:3915-3925, 1999; An, and Whitton, J. J. Virol. 71:2292,1997; Thomson, S. A. etal., J. Immunol. 157;822, 1996; Whitton, J. L etal., J. Virol.

67:348, 1993; Hanke, R. et al., Vaccine 16:426, 1998. For example, a multi-epitope DNA plasmid encoding supermotifand/or motif-bearing epitopes derived 109P1D4, the PADRE® universal helper T cell epitope or multiple HTL epitopes from 109P1 D4 (see Tables VIII-XXI and XXII to XLIX), and an endoplasmic reticulum-translocating signal sequence can be engineered. A vaccine may also comprise epitopes that are derived from other TAAs.

The immunogenicity of a multi-epitopic minigene can be confirmed in transgenlc mice to evaluate the magnitude of CTL induction responses against the epitopes tested. Further, the immunogenidty of DNA-encoded epitopes in vive can be correlated with the in vitro responses of specific CTL lines against target cells transfected with the DNA plasmid. Thus, these experiments can show that the minigene serves to both: generate a CTL response and that the induced CTLs recognized cells expressing the encoded epitopes.

For example, to create a DNA sequence encoding the selected epitopes (minigene) for expression in human cells, the amino acid sequences of the epitopes may be reverse translated. A human codon usage table can be used to guide the codon choice for each amino add. These epitope-encoding DNA sequences may be directly adjoined, so that when translated, a continuous polypeptide sequence is created. To optimize expression and/or Immunogenicity, additional elements can be incorporated into the minigene design. Examples of amino acid sequences that can be reverse translated and included in the minigene sequence include: HLA class I epitopes, HLA class II epitopes, antibody epitopes, a ubiquitination signal sequence, and/or an endoplasmic reticulum targeting signal. In addition, HLA presentation of CTL and HTL epltopes may be Improved by including synthetic poly-alanine) or naturally-occurring flanking sequences adjacent to the CTL or HTL epitopes; these larger peptides comprising the epitope(s) are within the scope of the invention.

The minigene sequence may be converted to DNA by assembling oligonucleotides that encode the plus and minus strands of the minigene. Overlapping oligonuceotides (30-100 bases long) may be synthesized, phosphorylated, purified 00 and annealed under appropriate conditions using well known techniques. The ends of the oligonucleotides can be joined, for 0example, using T4 DNA ligase. This synthetic minigene, encoding the epitope polypeptide, can then be cloned into a desired expression vector.

Standard regulatory sequences well known to those of skill in the art are preferably Included in the vector to ensure Sexpression in the target cells. Several vector elements are desirable: a promoter with a down-stream doning site for Sminigene insertion; a polyadenylation signal for efficient transcription termination; an E coliorigin of replication; and an E.

coli selectable marker ampicillin or kanamycin resistance). Numerous promoters can be used for this purpose, the human cytomegalovirus (hCMV) promoter. See, U.S. Patent Nos. 5,580,859 and 5,589,466 for other suitable promoter Ssequences.

SAdditional vector modifications may be desired to optimize minigene expression and immunogenicity. In some cases, Introns are required for efficient gene expression, and one or more synthetic or naturally-occurring introns could be C) incorporated into the transcribed region of the minigene. The inclusion of mRNA stabilization sequences and sequences for Sreplication in mammalian cells may also be considered for increasing minigene expression.

C Once an expression vector is selected, the minigene is cloned into the polylinker region downstream of the promoter. This plasmid is transformed into an appropriate E. co/i strain, and DNA Is prepared using standard techniques.

The orientation and DNA sequence of the minigene, as well as all other elements included in the vector, are confirmed using restriction mapping and DNA sequence analysis. Bacterial cells harboring the correct plasmid can be stored as a master cell bank and a working cell bank.

In addition, immunostimulatory sequences (ISSs or CpGs) appear to play a role in the immunogenidty of DNA vacdnes. These sequences may be included in the vector, outside the minigene coding sequence, if desired to enhance immunogenicity.

In some embodiments, a bi-cistronic expression vector which allows production of both the minigene-encoded epitopes and a second protein (included to enhance or decrease immunogenidty) can be used. Examples of proteins or polypeptides that could beneficially enhance the immune response if co-expressed include cytokines IL-2, IL-12, GM- CSF), cytokine-nducing molecules LelF), costimulatory molecules, or for HTL responses, pan-DR binding proteins

(PADRE

M

Epimmune, San Diego, CA). Helper (HTL) epitopes can be joined to intracellular targeting signals and expressed separately from expressed CTL epitopes; this allows direction of the HTL epitopes to a cell compartment different than that of the CTL epitopes. If required, this could facilitate more efficient entry of HTL epitopes into the HLA class II pathway, thereby improving HTL induction. In contrast to HTL or CTL induction, specifically decreasing the Immune response by co-expression of immunosuppressive molecules TGF-P) may be beneficial in certain diseases.

Therapeutic quantities of plasmid DNA can be produced for example, by fermentation in E co/i, followed by purification. Aliquots from the working cell bank are used to inoculate growth medium, and grown to saturation in shaker flasks or a bloreactor according to well-known techniques. Plasmid DNA can be purified using standard bloseparation technologies such as solid phase anion-exchange resins supplied by QIAGEN, Inc. (Valencia, California). If required, supercoiled DNA can be isolated from the open circular and linear forms using gel electrophoresis or other methods.

Purified plasmid DNA can be prepared for injection using a variety of formulations. The simplest of these is reconstitution of lyophilized DNA in sterile phosphate-buffer saline (PBS). This approach, known as "naked DNA," is currently being used for intramuscular (IM) administration in clinical trials. To maximize the Immunotherapeutic effects of minigene DNA vaccines, an altemative method for formulating purified plasmid DNA may be desirable. A variety of methods have been described, and new techniques may become available. Cationic lipids, glycolipids, and fusogenic liposomes can also be used in the formulation (see, as described by WO 93/24640; Mannino Gould-Fogerite, BloTechnlques 6(7): 00 682 (1988); U.S. Pat No. 5,279,833; WO 91/06309; and Feigner, et al, Proc. Nat'l Acad. Sci. USA 84:7413 (1987). In addition, peptides and compounds referred to collectively as protective, interactive, non-condensing compounds (PINC) C could also be complexed to purified plasmid DNA to Influence variables such as stability, intramuscular dispersion, or Strafficking to specific organs or cell types.

C) Target cell sensitization can be used as a functional assay for expression and HLA class I presentation of Smlnigene-encoded CTL epitopes. For example, the plasmid DNA is introduced into a mammalian cell line that is suitable as a target for standard CTL chromium release assays. The transfection method used will be dependent on the final formulation. Electroporation can be used for "naked" DNA, whereas cationic lipids allow direct in vitro transfection. A Splasmid expressing green fluorescent protein (GFP) can be co-transfected to allow enrichment of transfected cells using fluorescence activated cell sorting (FACS). These cells are then chromium-51 (51Cr) labeled and used as target cells for epitope-specific CTL ines; cytolysis, detected by 6 1 Cr release, indicates both production of, and HLA presentation of, Sminigene-encoded CTL epitopes. Expression of HTL epitopes may be evaluated In an analogous manner using assays to 00 0 assess HTL activity.

0 In viv immunogenicity is a second approach for functional testing of minigene DNA formulations. Transgenic mice expressing appropriate human HLA proteins are immunized with the DNA product The dose and route of administration are formulation dependent IM for DNA in PBS, intraperitoneal for lipid-complexed DNA). Twenty-one days after immunization, splenocytes are harvested and restimulated for one week in the presence of peptides encoding each epitope being tested. Thereafter, for CTL effector cells, assays are conducted for cytolysis of peptide-loaded, 5 lCr-labeled target cells using standard techniques. Lysis of target cells that were sensitized by HLA loaded with peptide epitopes, corresponding to minigene-encoded epitopes, demonstrates DNA vaccine function for in vivo induction of CTLs.

Immunogenicity of HTL epitopes is confirmed in transgenic mice in an analogous manner.

Alternatively, the nucleic acids can be administered using ballistic delivery as described, for instance, in U.S.

Patent No. 5,204,253. Using this technique, particles comprised solely of DNA are administered. In a further alternative embodiment, DNA can be adhered to partides, such as gold particles.

Minigenes can also be delivered using other bacterial or viral delivery systems well known In the art, an expression construct encoding epitopes of the invention can be incorporated into a viral vector such as vaccinia.

X.C2. Combinations of CTL Peptides with Helper Peptides Vaccine compositions comprising CTL peptides of the invention can be modified, analoged, to provide desired attributes, such as improved serum half life, broadened population coverage or enhanced immunogenicity.

For instance, the ability of a peptide to induce CTL activity can be enhanced by linking the peptide to a sequence which contains at least one epitope that is capable of inducing a T helper cell response. Although a CTL peptide can be directly linked to a T helper peptide, often CTL epitope/HTL epitope conjugates are linked by a spacer molecule. The spacer is typically comprised of relatively small, neutral molecules, such as amino acids or amino acid mimetics, which are substantially uncharged under physiological conditions. The spacers are typically selected from, Ala, Gly, or other neutral spacers of nonpolar amino acids or neutral polar amino adds. It will be understood that the optionally present spacer need not be comprised of the same residues and thus may be a hetero- or homo-oligomer. When present, the spacer will usually be at least one or two residues, more usually three to six residues and sometimes 10 or more residues. The CTL peptide epitope can be linked to the T helper peptide epitope either directly or via a spacer either at the amino or carboxy terminus of the CTL peptide. The amino terminus of either the immunogenic peptide or the T helper peptide may be acylated.

In certain embodiments, the T helper peptide is one that is recognized by T helper cells present in a majority of a 00 genetically diverse population. This can be accomplished by selecting peptides that bind to many, most, or all of the HLA Sclass II molecules. Examples of such amino acid bind many HLA Class II molecules include sequences from antigens such 1 as tetanus toxoid at positions 830-843 QYIKANSKFIGITE; (SEQ ID NO: 40), Plasmodium falclpatum circumsporozoite (CS) protein at positions 378-398 DIEKKIAKMEKASSVFNWNS; (SEQ ID NO: 41), and Streptococcus 18kD protein at positions 116-131 GAVDSILGGVATYGAA; (SEQ ID NO: 42). Other examples include peptides bearing a DR 1-4-7 supermotif, or l either of the DR3 motifs.

Altematively, it is possible to prepare synthetic peptides capable of stimulating T helper lymphocytes, in a loosely HLA-restricted fashion, using amino acid sequences not found in nature (see, PCT publication WO 95/07707). These Ssynthetic compounds called Pan-DR-binding epitopes PADRE T m Epimmune, Inc., San Diego, CA) are designed, most 0preferably, to bind most HLA-DR (human HLA class II) molecules. For instance, a pan-DR-binding epitope peptide having the formula: xKXVAAWTLKAAx (SEQ ID NO: 43), where is either cyclohexylalanine, phenylalanine, or tyrosine, and a is CN either D-alanine or L-alanine, has been found to bind to most HLA-DR alleles, and to stimulate the response of T helper 00 Slymphocytes from most individuals, regardless of their HLA type. An alternative of a pan-DR binding epitope comprises all S"L' natural amino acids and can be provided in the form of nucleic acids that encode the epitope.

HTL peptide epitopes can also be modified to alter their biological properties. For example, they can be modified to Include D-amino adds to increase their resistance to proteases and thus extend their serum half life, or they can be conjugated to other molecules such as lipids, proteins, carbohydrates, and the like to increase their biological activity. For example, a T helper peptide can be conjugated to one or more palmitic add chains at either the amino or carboxyl termini.

X.C.3. Combinations of CTL Peptides with T Cell Priming Agents In some embodiments it may be desirable to include in the pharmaceutical compositions of the invention at least one component which primes B lymphocytes or T lymphocytes. Uplds have been identified as agents capable of priming CTL in vivo. For example, palmitic add residues can be attached to the e-and a- amino groups of a lysine residue and then linked, via one or more linking residues such as Gly, Gly-Gly-, Ser, Ser-Ser, or the like, to an immunogenic peptide.

The lipldated peptide can then be administered either directly in a micelle or particle, incorporated Into a liposome, or emulsified in an adjuvant, incomplete Freund's adjuvant In a preferred embodiment, a particularly effective immunogenic composition comprises palmitic add attached to e- and a- amino groups of Lys, which is attached via linkage, Ser-Ser, to the amino terminus of the immunogenic peptide.

As another example of lipid priming of CTL responses, E coli lipoproteins, such as tripalmltoyl-Sglycerylcysteinlyseryl- serine (PaCSS) can be used to prime virus specific CTL when covalently attached to an appropriate peptide (see, Deres, etal., Nature 342561, 1989). Peptides of the invention can be coupled to P3CSS, for example, and the lipopeptide administered to an individual to prime specifically an immune response to the target antigen. Moreover, because the induction of neutralizing antibodies can also be primed with P3CSS-conjugated epitopes, two such compositions can be combined to more effectively elicit both humoral and cell-mediated responses.

XC.4. Vaccine Compositions Comprising DC Pulsed with CTL andlor HTL Peptides An embodiment of a vaccine composition in accordance with the invention comprises ex vivo administration of a cocktail of epitope-bearing peptides to PBMC, or Isolated DC therefrom, from the patient's blood. A pharmaceutical to facilitate harvesting of DC can be used, such as Progenipoietin M (Pharmacla-Monsanto, St Louis, MO) or GM-CSFIL-4.

After pulsing the DC with peptides and prior to reinfuslon Into patients, the DC are washed to remove unbound peptides. In this embodiment, a vaccine comprises peptide-pulsed DCs which present the pulsed peptide epitopes complexed with HLA molecules on their surfaces.

The DC can be pulsed ex vivo with a cocktail of peptides, some of which stimulate CTL responses to 109P1 D4.

0 Optionally, a helper T cell (HTL) peptide, such as a natural or artificial loosely restricted HLA Class II peptide, can be 0included to facilitate the CTL response. Thus, a vaccine in accordance with the invention is used to treat a cancer which expresses or overexpresses 109P1D4.

X.D. Adoptive Immunotherapy Antigenic 109P1 D4-related peptides are used to elicit a CTL and/or HTL response ex vivo, as well. The resulting CTL or HTL cells, can be used to treat tumors in patients that do not respond to other conventional forms of therapy, or will Snot respond to a therapeutic vaccine peptide or nudeic acid in accordance with the invention. Ex vivo CTL or HTL Cl responses to a particular antigen are Induced by incubating in tissue culture the patients, or genetically compatible, CTL or 0' HTL precursor cells together with a source of antigen-presenting cells (APC), such as dendritic cells, and the appropriate immunogenic peptide. After an appropriate Incubation time (typically about 7-28 days), in which the precursor cells are 00 activated and expanded into effector cells, the cells are infused back into the patient, where they will destroy (CTL) or Sfacilitate destruction (HTL) of their specific target cell a tumor cell). Transfected dendritic cells may also be used as CN antigen presenting cells.

X.E. Administration of Vaccines for Therapeutic or Prophylactic Purposes Pharmaceutical and vaccine compositions of the invention are typically used to treat and/or prevent a cancer that expresses or overexpresses 109P1D4. In therapeutic applications, peptide and/or nucleic add compositions are administered to a patient in an amount sufficient to elicit an effective B cell, CTL and/or HTL response to the antigen and to Scure or at least partially arrest or slow symptoms and/or complications. An amount adequate to accomplish this is defined as "therapeutically effective dose." Amounts effective for this use will depend on, the particular composition administered, the manner of administration, the stage and severity of the disease being treated, the weight and general state of health of the patient, and the judgment of the prescribing physician.

For pharmaceutical compositions, the Immunogenic peptides of the invention, or DNA encoding them, are generally administered to an Individual already bearing a tumor that expresses 109P1D4. The peptides or DNA encoding them can be administered individually or as fusions of one or more peptide sequences. Patients can be treated with the immunogenic peptides separately or in conjunction with other treatments, such as surgery, as appropriate.

For therapeutic use, administration should generally begin at the first diagnosis of 109P1D4-associated cancer.

This is followed by boosting doses until at least symptoms are substantially abated and for a period thereafter. The embodiment of the vaccine composition including, but not limited to embodiments such as peptide cocktails, polyepitopic polypeptides, minigenes, or TAA-specific CTLs or pulsed dendritic cells) delivered to the patient may vary according to the stage of the disease or the patient's health status. For example, in a patient with a tumor that expresses 109P1 D4, a vaccine comprising 109P1 D4-specific CTL may be more efficacious in killing tumor cells in patient with advanced disease than altemative embodiments.

It is generally important to provide an amount of the peptide epitope delivered by a mode of administration sufficient to stimulate effectively a cytotoxic T cell response; compositions which stimulate helper T cell responses can also be given in accordance with this embodiment of the invention.

The dosage for an initial therapeutic immunization generally occurs In a unit dosage range where the lower value is about 1, 5, 50, 500, or 1,000 pg and the higher value is about 10,000; 20,000; 30,000; or 50,000 pg. Dosage values for a human typically range from about 500 pg to about 50,000 pg per 70 kilogram patient Boosting dosages of between about pg to about 50,000 pg of peptide pursuant to a boosting regimen over weeks to months may be administered depending 00 upon the patients response and condition as determined by measuring the specific activity of 0Th and HTL obtained from 0 the patient's blood. Administration should continue until at least clinical symptoms or laboratory tests indicate that the neoplasia, has been eliminated or reduced and for a period thereafter. The dosages, routes of administration, and dose schedules are adjusted In accordance with methoologies known in the art In certain embodiments, the peptides and compositions of the present Invention are employed in serious disease states, that is, life-threatening or potentially life threatening situations. In such cases, as a result of the minimal amounts of extraneous substances and the relative nontoxic nature of the peptides in preferred compositions of the invention, It is possible and may be felt desirable by the treating physician to administer substantial excesses of these peptide compositions c-i relative to these stated dosage amounts.

The vaccine compositions of the invention can also be used purely as prophylactic agents. Generally the dosage for an initial prophylactic immunization generally occurs In a unit dosage range where the lower value Is about 1, 5, 50, 500, 00 or 1000 pg and the higher value Is about 10,000; 20,000; 30,000; or 50,000 pg. Dosage values for a human typically range from about 500 pg to about 50,000 pg per 70 kilogram patient This Is followed by boosting dosages of between about 1 .0 pig to about 50,000 jig of peptide administered at defined intervals from about four weeks to six months after the initial administration of vaccine. The immunogenicity of the vaccine can be assessed by measuring the specific activity of CTL and Ht obtained from a sample of the patient's blood.

The pharmaceutical compositions for therapeutic treatment are Intended for parenteral, topical, oral, nasal, intrathecal, or local as a cream or topical ointment) administration. Preferably, the pharmaceutical compositions are administered parentally, intravenously, subcutaneously, intradermally, or intramuscularly. Thus, the invention provides compositions for parenteral administration which comprise a solution of the immunogenic peptides dissolved or suspended In an acceptable carrier, preferably an aqueos carder.

A variety of aqueous carriers may be used, water, buffered water, 0.8% saline, 0.3% glycine, hyaluronic acd and the like. These compositions may be sterilized by conventional, well-known sterilizaton techniques, or may be sterile filtered. The resulting aqueous solutions may be packaged for use as is, or lyophilized, the tyophilized preparation being combined with a sterile solution prior to administration.

The ccmpositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions, such as pl-1-adjusting and buffering agents, tonicity adjusting agents, wetting agents, preservatives, and the like, for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate, triethanolamine oleate, etc.

The concentration of peptides of the invention In the pharmaceutical formulations can vary widely, iLe., from less than about 0.1 usually at or at least about 2% to as much as 20% to 50% or more by weight~ and will be selected primarily by fluid volumes, viscosities, etc., in accordance with the particular mode of administration selected.

A human unit dose form of a composition is typically Included In a pharmaceutical composition that comprises a human unit dose of an acceptable cardier, in one embodiment an aqueous carrier, and Is administered in a volumelquantity that Is known by those of skill In the art to be used for administration of such compositions to humans (see, Remington's Pharmaceutical Sciences, 17th Edition, A. Gennaro, Editor, Mack Publishing Co., Easton, Pennsylvania, 1985). For example a peptide dose for initial Immunization can be from about 1 to about 50,000 pig, generally 100-5,000 pig, for a 70 kg patient For example, for nucleic acids an initial Immunization may be performed using an exprssion vector in the form of naked nucleic acid administered IM (or SC or ID) In the amounts of 0.5-5 mg at multiple sites. The nuclec acdd (0.1 to 1000 pig) can also be administered using a gene gun. Following an incubation period of 3-4 weeks, a booster dose Is then administered. The booster can be recombinant fowlpox: virus administered at a dose of 5-107 to 5x10 9 pfu.

0 0 For antibodies, a treatment generally involves repeated administration of the anti-109P1D4 antibody preparation, via an acceptable route of administration such as intravenous injection typically at a dose in the range of about 0.1 to about 10 mg/kg body weight In general, doses in the range of 10-500 mg mAb per week are effective and well tolerated.

Moreover, an initial loading dose of approximately 4 mg/kg patient body weight IV, followed by weekly doses of about 2 Smg/kg IV of the ani- 109P1D4 mAb preparation represents an acceptable dosing regimen. As appreciated by those of skill in the art, various factors can influence the ideal dose in a particular case. Such factors include, for example, half life of a composition, the binding affinity of an Ab, the immunogenicity of a substance, the degree of 109P1 4 expression in the patient, the extent of circulating shed 109P1D4 antigen, the desired steady-state concentration level, frequency of treatment, Sand the influence of chemotherapeutic or other agents used In combination with the treatment method of the invention, as Swell as the health status of a particular patient Non-limiting preferred human unit doses are, for example, 500pg img, img 50mg, 50mg 100mg, 100mg 200mg, 200mg 300mg, 400mg 500mg, 500mg 600mg, 60 0 mg 700mg, 700mg 800mg, 800mg 900mg, 900mg ig, or 1mg 700mg. In certain embodiments, the dose is in a range of 2-5 mg/kg body 00 O weight, with follow on weekly doses of 1-3 mg/kg; 0.5mg, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10mg/kg body weight followed, in two, three or four weeks by weekly doses; 0.5 10mg/kg body weight followed in two, three or four weeks by weekly doses; 225, 250, 275, 300, 325, 350, 375, 400mg m 2 of body area weekly; 1-600mg m 2 of body area weekly; 225-400mg m 2 of body area weekly; these does can be followed by weeldy doses for 2, 3, 4, 5, 6, 7, 8, 9, 19, 11, 12 or more weeks.

In one embodiment, human unit dose forms of polynucleotides comprise a suitable dosage range or effective amount that provides any therapeutic effect As appreciated by one of ordinary skill in the art a therapeutic effect depends on a number of factors, induding the sequence of the polynuceotide, molecular weight of the polynudeotide and route of administration. Dosages are generally selected by the physician or other health care professional in accordance with a variety of parameters known in the art, such as severity of symptoms, history of the patient and the like. Generally, for a polynudeotide of about 20 bases, a dosage range may be selected from, for example, an independently selected lower limit such as about 0.1, 0.25, 0.5, 1, 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400 or 500 mg/kg up to an independently selected upper limit, greater than the lower limit, of about 60, 80, 100, 200, 300, 400, 500, 750, 1000, 1500, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000 or 10,000 mg/kg. For example, a dose may be about any of the following: 0.1 to 100 mg/kg, 0.1 to 50 mg/kg, 0.1 to 25 mg/kg, 0.1 to 10 mg/kg, 1 to 500 mg/kg, 100 to 400 mg/kg, 200 to 300 mg/kg, 1 to 100 mg/kg, 100 to 200 mg/kg, 300 to 400 mg/kg, 400 to 500 mg/kg, 500 to 1000 mg/kg, 500 to 5000 mg/kg, or 500 to 10,000 mg/kg. Generally, parenteral routes of administration may require higher doses of polynudeotide compared to more direct application to the nucleotide to diseased tissue, as do polynudeotides of increasing length.

In one embodiment, human unit dose forms of T-cells comprise a suitable dosage range or effective amount that provides any therapeutic effect. As appreciated by one of ordinary skill in the art, a therapeutic effect depends on a number of factors. Dosages are generally selected by the physician or other health care professional In accordance with a variety of parameters known in the art, such as severity of symptoms, history of the patient and the like. A dose may be about 104 cells to about 106 cells, about 106 cells to about 10 8 cells, about 108 to about 1011 cells, or about 108 to about 5 x 10 o cells.

A dose may also about 106 cells/m 2 to about 101o cells/nm, or about 106 cellsm2 to about 108 cells/m 2 Proteins(s) of the Invention, and/or nucleic acids encoding the protein(s), can also be administered via liposomes, which may also serve to: 1) target the proteins(s) to a particular tissue, such as lymphoid tissue; 2) to target selectively to diseases cells; or, 3) to increase the half-life of the peptide composition. Uposomes include emulsions, foams, micelles, insoluble monolayers, liquid crystals, phospholipid dispersions, lamellar layers and the like. In these preparations, the peptide to be delivered is Incorporated as part of a liposome, alone or in conjunction with a molecule which binds to a receptor prevalent among lymphoid cells, such as monodonal antibodies which bind to the CD45 antigen, or with other 64 00 therapeutic or immunogenic compositions. Thus, liposomes either filled or decorated with a desired peptide of the invention can be directed to the site of lymphoid cells, where the liposomes then deliver the peptide compositions. Lposomes for use Sin accordance with the invention are formed from standard vesice-forming lipids, which generally include neutral and negatively charged phospholpids and a sterol, such as cholesterol. The selection of lipids is generally guided by 7) consideration of, liposome size, add lability and stability of the liposomes in the blood stream. A variety of methods are Savailable for preparing liposomes, as described in, Szoka, et al., Ann. Rev. Biophys. Bioeng. 9:467 (1980), and U.S.

Patent Nos. 4,235,871, 4,501,728, 4,837,028, and 5,019,369.

For targeting cells of the immune system, a ligand to be incorporated into the lposome can include, e.g., Santibodies or fragments thereof specific for cell surface determinants of the desired immune system cells. A liposome Ssuspension containing a peptide may be administered intravenously, locally, topically, etc. in a dose which varies according to, Inter alia, the manner of administration, the peptide being delivered, and the stage of the disease being treated.

ri For solid compositions, conventional nontoxic solid carriers may be used which indude, for example, 00 0pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, cellulose, glucose, Ssucrose, magnesium carbonate, and the like. For oral administration, a pharmaceutically acceptable nontoxic composition is formed by incorporating any of the normally employed exdplents, such as those carriers previously listed, and generally of active Ingredient, that is, one or more peptides of the invention, and more preferably at a concentration of 25%-75%.

For aerosol administration, immunogenic peptides are preferably supplied in finely divided form along with a surfactant and propellant Typical percentages of peptides are about 0.01%-20% by weight, preferably about 1 The surfactant must, of course, be nontoxic, and preferably soluble in the propellant Representative of such agents are the esters or partial esters of fatty adds containing from about 6 to 22 carbon atoms, such as caproic, octanoic, lauric, palmitic, stearic, linoleic, linolenlc, olesteric and oleic adds with an aliphatic polyhydric alcohol or Its cyclic anhydride. Mixed esters, such as mixed or natural glycerides may be employed. The surfactant may constitute about 0.1%-20% by weight of the composition, preferably about 0.25-5%. The balance of the composition is ordinarily propellant A carrier can also be included, as desired, as with, lecithin for intranasal delivery.

XL) Diagnostic and Proanostic Embodiments of 109P1D4.

As disclosed herein, 109P1D4 polynucleotides, polypeptides, reactive cytotoxic T cells (CTL), reactive helper T cells (HTL) and anti-polypeptide antibodies are used in well known diagnostic, prognostic and therapeutic assays that examine conditions associated with dysregulated cell growth such as cancer, in particular the cancers listed in Table I (see, both its specific pattern of tissue expression as well as its overexpression in certain cancers as described for example In the Example entitled 'Expression analysis of 109P1D4 In normal tissues, and patient specimens).

109P1D4 can be analogized to a prostate associated antigen PSA, the archetypal marker that has been used by medical practitioners for years to identify and monitor the presence of prostate cancer (see, Merrill et al., J. Urol. 163(2): 503-5120 (2000); Polasdk etal., J. Urol. Aug; 162(2):293-306 (1999) and Fortier et al., J. Nat Cancer Inst 91(19): 1635- 1640(1999)). A variety of other diagnostic markers are also used In similar contexts including p53 and K-ras (see, e.g., Tulchinsky et al., Int J Mol Med 1999 Jul 4(1):99-102 and Minlmoto et al., Cancer Detect Prey 2000;24(1):1-12). Therefore, this disclosure of 109P1D4 polynuceotides and polypeptides (as well as 109P1D4 polynudeotide probes and anti-109P1D4 antibodies used to identify the presence of these molecules) and their properties allows skilled artisans to utilize these molecules in methods that are analogous to those used, for example, in a variety of diagnostic assays directed to examining conditions associated with cancer.

Typical embodiments of diagnostic methods which utilize the 109P1D4 polynudeotides, polypeptides, reactive T 00 cells and antibodies are analogous to those methods from well-established diagnostic assays, which employ, PSA 0 polynudeotides, polypeptides, reactive T cells and antibodies. For example, just as PSA polynuceotides are used as probes (for example in Northern analysis, see, Sharief et al., Biochem. Mol. Biol. Int 33(3):567-74(1994)) and primers (for L) example in PCR analysis, see, Okegawa et al., J. Urol. 163(4): 1189-1190 (2000)) to observe the presence and/or the Slevel of PSA mRNAs in methods of monitoring PSA overexpression or the metastasis of prostate cancers, the 109P1D4 Spolynudeotides described herein can be utilized In the same way to detect 109P1D4 overexpression or the metastasis of prostate and other cancers expressing this gene. Altematively, just as PSA polypeptides are used to generate antibodies specific for PSA which can then be used to observe the presence and/or the level of PSA proteins in methods to monitor SPSA protein overexpression (see, Stephan et al., Urology 55(4):560-3 (2000)) or the metastasis of prostate cells (see, Se.g., Alanen et al., Pathol. Res. Pract 192(3):233-7 (1996)), the 109P1D4 polypeptides described herein can be utilized to generate antibodies for use in detecting 109P1D4 overexpression or the metastasis of prostate cells and cells of other OC cancers expressing this gene.

SSpecifically, because metastases involves the movement of cancer cells from an organ of origin (such as the lung or prostate gland etc.) to a different area of the body (such as a lymph node), assays which examine a biological sample for the presence of cells expressing 109P1 D4 polynucleotides and/or polypeptides can be used to provide evidence of metastasis. For example, when a biological sample from tissue that does not normally contain 109P1D4-expressing cells (lymph node) is found to contain 109P1D4-expressing cells such as the 109P1D4 expression seen in LAPC4 and LAPC9, xenografts isolated from lymph node and bone metastasis, respectively, this finding is indicative of metastasis.

Alternatively 109P1D4 polynudeotides and/or polypeptides can be used to provide evidence of cancer, for example, when cells in a biological sample that do not normally express 109P1 D4 or express 109P1 D4 at a different level are found to express 109P1D4 or have an increased expression of 109P1D4 (see, the 109P1D4 expression in the cancers listed in Table I and in patient samples etc. shown in the accompanying Figures). In such assays, artisans may further wish to generate supplementary evidence of metastasis by testing the biological sample for the presence of a second tissue restricted marker (in addition to 109P1D4) such as PSA, PSCA etc. (see, Alanen et al., Pathol. Res. Pract 192(3): 233- 237 (1996)).

The use of immunohistochemistry to identify the presence of a 109P1D4 polypeptide within a tissue section can indicate an altered state of certain cells within that tissue. It is well understood in the art that the ability of an antibody to localize to a polypeptide that is expressed in cancer cells is a way of diagnosing presence of disease, disease stage, progression and/or tumor aggressiveness. Such an antibody can also detect an altered distribution of the polypeptide within the cancer cells, as compared to corresponding non-malignant tissue.

The 109P1D4 polypeptide and immunogenic compositions are also useful in view of the phenomena of altered subcellular protein localization In disease states. Alteration of cells from normal to diseased state causes changes in cellular morphology and is often associated with changes in subcellular protein localizationldistribution. For example, cell membrane proteins that are expressed in a polarized manner in normal cells can be altered in disease, resulting In distribution of the protein in a non-polar manner over the whole cell surface.

The phenomenon of altered subcellular protein localization in a disease state has been demonstrated with MUC1 and Her2 protein expression by use of immunohistochemical means. Normal epithelial cells have a typical apical distribution of MUC1, in addition to some supranudear localization of the glycoprotein, whereas malignant lesions often demonstrate an apolar staining pattern (Diaz et al, The Breast Journal, 7; 40-45 (2001); Zhang et al, Clinical Cancer Research, 4; 2669-2676 (1998): Cao, et al, The Journal of Histochemlstry and Cytochemistry, 45:1547-1557 (1997)). In addition, normal breast epithelium Is either negative for Her2 protein or exhibits only a basolateral distribution whereas malignant cells can express

I

00 the protein over the whole cell surface (De Potter, et al, International Journal of Cancer, 44; 969-974 (1989): McCormick, et al, 117; 935-943 (2002)). Alternatively, distribution of the protein may be altered from a surface only localization to include diffuse cytoplasmic expression In the diseased state. Such an example can be seen with MUC1 (Diaz, et al, The Breast S Journal, 7:40-45 (2001)).

SAlteration in the localizationdistribution of a protein in the cell, as detected by Immunohistochemical methods, can S also provide valuable information concerning the favorability of certain treatment modalities. This last point is illustrated by a situation where a protein may be intracellular in normal tissue, but cell surface In malignant cells; the cell surface location makes the cells favorably amenable to antibody-based diagnostic and treatment regimens. When such an alteration of Sprotein localization occurs for 109P1 D4, the 109P1D4 protein and immune responses related thereto are very useful.

SAccordingly, the ability to determine whether alteration of subcellular protein localization occurred for 24P4C12 make the 109P1D4 protein and immune responses related thereto very useful. Use of the 109P1D4 compositions allows those skilled in the art to make important diagnostic and therapeutic decisions.

SImmunohistochemlcal reagents specific to 109P1D4 are also useful to detect metastases of tumors expressing 109P1D4 C when the polypeptide appears in tissues where 109P1D4 is not normally produced.

Thus, 109P1 D4 polypeptides and antibodies resulting from immune responses thereto are useful in a variety of Important contexts such as diagnostic, prognostic, preventative and/or therapeutic purposes known to those skilled in the art Just as PSA polynucleotide fragments and polynudeotide variants are employed by skilled artisans for use in methods of monitoring PSA, 109P1D4 polynucleotide fragments and polynudeotide variants are used In an analogous manner. In particular, typical PSA polynudeotides used in methods of monitoring PSA are probes or primers which consist of fragments of the PSA cONA sequence. Illustrating this, primers used to PCR amplify a PSA polynudeotide must include less than the whole PSA sequence to function in the polymerase chain reaction. In the context of such PCR reactions, skilled artisans generally create a variety of different polynudeotide fragments that can be used as primers in order to amplify different portions of a polynudeotide of interest or to optimize amplification reactions (see, Caetano-Anolles, G.

Biotechniques 25(3): 472-476,478-480 (1998); Robertson etal., Methods Mol. Biol. 98:121-154 (1998)). An additional illustration of the use of such fragments is provided In the Example entitled "Expression analysis of 109P1D4 In normal tissues, and patient specimens," where a 109P1D4 polynudeotide fragment is used as a probe to show the expression of 109P1D4 RNAs in cancer cells. In addition, variant polynucleotide sequences are typically used as primers and probes for the corresponding mRNAs in PCR and Northern analyses (see, Sawai et al., Fetal Diagn. Ther. 1996 Nov-Dec 11(6):407-13 and Current Protocols In Molecular Biology, Volume 2, Unit 2, Frederick M. Ausubel et al eds., 1995)).

Polynudeotide fragments and variants are useful in this context where they are capable of binding to a target polynuceotide sequence a 109P1D4 polynucleotide shown in Figure 2 or variant thereof) under conditions of high stringency.

Furthermore, PSA polypeptides which contain an epitope that can be recognized by an antibody or T cell that specifically binds to that epitope are used in methods of monitoring PSA. 109P104 polypeptide fragments and polypeptide analogs or variants can also be used in an analogous manner. This practice of using polypeptide fragments or polypeptide variants to generate antibodies (such as anti-PSA antibodies or T cells) is typical in the art with a wide variety of systems such as fusion proteins being used by practitioners (see, Current Protocols In Molecular Biology, Volume 2, Unit 16, Frederick M. Ausubel etal. eds., 1995). In this context, each epitope(s) functions to provide the architecture with which an antibody or T cell is reactive. Typically, skilled artisans create a variety of different polypeptide fragments that can be used in order to generate Immune responses specific for different portions of a polypeptide of interest (see, U.S. Patent No.

5,840,501 and U.S. Patent No. 5,939,533). For example it may be preferable to utilize a polypeptide comprising one of the 109P1D4 biological motifs discussed herein or a motif-bearing subsequence which is readily Identified by one of skill in the 0 art based on motifs available in the art. Polypeptide fragments, variants or analogs are typically useful in this context as long Sas they comprise an epitope capable of generating an antibody or T cell specific for a target polypeptide sequence a 109P1D4 polypeptide shown in Figure 3).

As shown herein, the 109P1D4 polynuceotides and polypeptides (as well as the 109P1 D4 polynudeotide probes and anti-109P D4 antibodies or T cells used to identify the presence of these molecules) exhibit specific properties that 'make them useful in diagnosing cancers such as those listed in Table I. Diagnostic assays that measure the presence of 109P1D4 gene products, in order to evaluate the presence or onset of a disease condition described herein, such as prostate cancer, are used to identify patients for preventive measures or further monitoring, as has been done so Ssuccessfully with PSA. Moreover, these materials satisfy a need In the art for molecules having similar or complementary Scharacteristics to PSA in situations where, for example, a definite diagnosis of metastasis of prostatic origin cannot be made on the basis of a test for PSA alone (see, Alanen et al., Pathol. Res. Pract 192(3): 233-237 (1996)), and consequently, 00 materials such as 109P1D4 polynudeotides and polypeptides (as well as the 109P1D4 polynucleotide probes and anti- O 109P1D4 antibodies used to identify the presence of these molecules) need to be employed to confirm a metastases of Cl prostatic origin.

Finally, in addition to their use in diagnostic assays, the 109P1D4 polynucleotides disclosed herein have a number of other utilities such as their use in the identification of oncogenetic associated chromosomal abnormalities In the chromosomal region to which the 109P1D4 gene maps (see the Example entitled "Chromosomal Mapping of 109P1D4" below). Moreover, in addition to their use in diagnostic assays, the 109P1D4-related proteins and polynudeotides disdosed herein have other utilities such as their use in the forensic analysis of tissues of unknown origin (see, Takahama K Forensic Sd Int 1996 Jun 28;80(1-2): 63-9).

Additionally, 109P1D4-related proteins or polynudeotides of the invention can be used to treat a pathologic condition characterized by the over-expression of 109P1D4. For example, the amino acid or nucleic add sequence of Figure 2 or Figure 3, or fragments of either, can be used to generate an immune response to a 109P1D4 antigen. Antibodies or other molecules that react with 109P1 D4 can be used to modulate the function of this molecule, and thereby provide a therapeutic benefit XII.) Inhibition of 109P1 D4 Protein Function The Invention includes various methods and compositions for inhibiting the binding of 109P1D4 to its binding partner or its association with other protein(s) as well as methods for inhibiting 109P1D4 function.

XII.A Inhibition of 109P1D4 With Intracellular Antibodies In one approach, a recombinant vector that encodes single chain antibodies that specifically bind to 109P1D4 are introduced into 109P1D4 expressing cells via gene transfer technologies. Accordingly, the encoded single chain anti- 109P1D4 antibody is expressed intracellularly, binds to 109P1D4 protein, and thereby inhibits its function. Methods for engineering such intracellular single chain antibodies are well known. Such intracellular antibodies, also known as Intrabodies", are specifically targeted to a particular compartment within the cell, providing control over where the inhibitory activity of the treatment is focused. This technology has been successfully applied in the art (for review, see Richardson and Marasco, 1995, TIBTECH vol. 13). Intrabodies have been shown to virtually eliminate the expression of otherwise abundant cel surface receptors (see, Richardson et al., 1995, Proc. Natl. Acad. Sd. USA 92: 3137-3141; Beerli et al., 1994, J.

Biol. Chem. 289: 23931-23936; Deshane et al., 1994, Gene Ther. 1: 332-337).

Single chain antibodies comprise the variable domains of the heavy and light chain joined by a flexible linker 68 0 0 polypeptide, and are expressed as a single polypeptide. Optionally, single chain antibodies are expressed as a single chain variable region fragment joined to the light chain constant region. Well-known intracellular trafficking signals are engineered into recombinant polynudeotide vectors encoding such single chain antibodies in order to target precisely the intrabody to d) the desired intracellular compartment For example, intrabodies targeted to the endoplasmic reticulum (ER) are engineered Sto incorporate a leader peptide and, optionally, a C-terminal ER retention signal, such as the KDEL amino acid motif.

Intrabodies intended to exert activity In the nucleus are engineered to Indude a nuclear localization signal. Upld moieties are joined to intrabodies In order to tether the intrabody to the cytosolic side of the plasma membrane. Intrabodies can also be targeted to exert function in the cytosol. For example, cytosolic intrabodies are used to sequester factors within the cytosol, LC thereby preventing them from being transported to their natural cellular destination.

SIn one embodiment, intrabodies are used to capture 109P1D4 in the nucleus, thereby preventing its activity within the nucleus. Nuclear targeting signals are engineered into such 109P1D4 intrabodies in order to achieve the desired 0 targeting. Such 109P1D4 intrabodies are designed to bind specifically to a particular 109P1D4 domain. In another Sembodiment, cytosolic intrabodies that specifically bind to a 109P1 D4 protein are used to prevent 109P1 D4 from gaining Saccess to the nucleus, thereby preventing it from exerting any biological activity within the nucleus preventing 109P1D4 from forming transcription complexes with other factors).

In order to specifically direct the expression of such intrabodies to particular cells, the transcription of the intrabody is placed under the regulatory control of an appropriate tumor-specific promoter and/or enhancer. In order to target intrabody expression specifically to prostate, for example, the PSA promoter and/or promoter/enhancer can be utilized (See, for example, U.S. Patent No. 5,919,652 Issued 6 July 1999).

XII.B.) Inhibition of 109P1D4 with Recombinant Proteins In another approach, recombinant molecules bind to 109P1D4 and thereby Inhibit 109P1D4 function. For example, these recombinant molecules prevent or inhibit 109P1 D4 from accessing/binding to its binding partner(s) or associating with other protein(s). Such recombinant molecules can, for example, contain the reactive part(s) of a 109P1 D4 specific antibody molecule. In a particular embodiment the 109P1D4 binding domain of a 109P1D4 binding partner is engineered into a dimeric fusion protein, whereby the fusion protein comprises two 109P1 D4 ligand binding domains linked to the Fc portion of a human IgG, such as human IgG1. Such IgG portion can contain, for example, the CH2 and CH 3 domains and the hinge region, but not the CH1 domain. Such dimeric fusion proteins are administered in soluble form to patients suffering from a cancer associated with the expression of 109P D4, whereby the dimeric fusion protein specifically binds to 109P1 D4 and blocks 109P1D4 interaction with a binding partner. Such dimeric fusion proteins are further combined into multimeric proteins using known antibody linking technologies.

XII.C.) Inhibition of 109P D4 Transcription or Translation The present Invention also comprises various methods and compositions for inhibiting the transcription of the 109P1D4 gene. Similarly, the invention also provides methods and compositions for inhibiting the translation of 109P1D4 mRNA into protein.

In one approach, a method of inhibiting the transcription of the 109P1D4 gene comprises contacting the 109P1D4 gene with a 109PiD4 antisense polynudeotide. In another approach, a method of inhibiting 109P1D4 mRNA translation comprises contacting a 109P1D4 mRNA with an antisense polynudeotide. In another approach, a 109P1 D4 specific ribozyme is used to cleave a 109P1D4 message, thereby inhibiting translation. Such antisense and ribozyme based methods can also be directed to the regulatory regions of the 109P1D4 gene, such as 109P1 4 promoter and/or enhancer 69 00 elements. Similarly, proteins capable of inhibiting a 109P1D4 gene transcription factor are used to inhibit 109P1D4 mRNA transcription. The various polynucleotides and compositions useful in the aforementioned methods have been described

C

above. The use of antisense and ribozyme molecules to inhibit transcription and translation Is well known in the art 0) Other factors that inhibit the transcription of 109P1 D4 by interfering with 109P1D4 transcriptional activation are also useful to treat cancers expressing 109P1D4. Similarly, factors that interfere with 109P1D4 processing are useful to treat V' cancers that express 109P1 D4. Cancer treatment methods utilizing such factors are also within the scope of the invention.

XIID.) General Considerations for Theraneutic Strateies SGene transfer and gene therapy technologies can be used to deliver therapeutic polynucleotide molecules to tumor cells Ssynthesizing 109P1D4 antisense, ribozyme, polynudeotides encoding intrabodies and other 109P1D4 inhibitory molecules).

S A number of gene therapy approaches are known in the art Recombinant vectors encoding 109P1D4 antisense polynudeotides, n ibozymes, factors capable of interfering with 109P1D4 transcription, and so forth, can be delivered to target tumor cells using Ssuch gene therapy approaches.

0 The above therapeutic approaches can be combined with any one of a wide variety of surgical, chemotherapy or radiation therapy regimens. The therapeutic approaches of the invention can enable the use of reduced dosages of chemotherapy (or other therapies) and/or less frequent administration, an advantage for all patients and particularly for those that do not tolerate the toxicity of the chemotherapeutic agent well.

The anti-tumor activity of a particular composition antisense, ribozyme, intrabody), or a combination of such compositions, can be evaluated using various in vitro and in viv assay systems. In vtro assays that evaluate therapeutic activity indude cell growth assays, soft agar assays and other assays indicative of tumor promoting activity, binding assays capable of determining the extent to which a therapeutic composition will inhibit the binding of 109P1 D4 to a binding partner, etc.

In vivo, the effect of a 109P1D4 therapeutic composition can be evaluated in a suitable animal model. For example, xenogenic prostate cancer models can be used, wherein human prostate cancer explants or passaged xenograft tissues are introduced into immune compromised animals, such as nude or SCID mice (Klein et at., 1997, Nature Medicine 3: 402-408). For example, PCT Patent Application W098/16628 and U.S. Patent 6,107,540 describe various xenograft models of human prostate cancer capable of recapitulating the developrment of primary tumors, micrometastasis, and the formation of osteoblastic metastases characteristic of late stage disease. Efficacy can be predicted using assays that measure Inhibition of tumor formation, tumor regression or metastasis, and the like.

In vivo assays that evaluate the promotion of apoptosis are useful in evaluating therapeutic compositions. In one embodiment xenografts from tumor bearing mice treated with the therapeutic composition can be examined for the presence of apoptotic foci and compared to untreated control xenograft-bearing mice. The extent to which apoptotic foci are found in the tumors of the treated mice provides an Indication of the therapeutic efficacy of the composition.

The therapeutic compositions used in the practice of the foregoing methods can be formulated into pharmaceutical compositions comprising a carrier suitable for the desired delivery method. Suitable carriers include any material that when combined with the therapeutic composition retains the anti-tumor function of the therapeutic composition and Is generally non-reactive with the patients immune system. Examples include, but are not limited to, any of a number of standard pharmaceutical carriers such as stenle phosphate buffered saline solutions, bacteriostatic water, and the like (see, generally, Remington's Pharmaceutical Sciences 16d Edition, A. Osal., Ed., 1980).

Therapeutic formulations can be solubilized and administered via any route capable of delivering the therapeutic composition to the tumor site. Potentially effective routes of administration indude, but are not limited to, intravenous, parenteral, intraperitoneal, Intramuscular, intratumor, intradermal, intraorgan, orthotopic, and the like. A preferred 00 formulation for intravenous injecton comprises the therapeutic composition in a solution of preserved bacteriostatic water, sterile unpreserved water, and/or diluted In polyvinylchloride or polyethylene bags containing 0.9% sterile Sodium Chloride for Injection, USP. Therapeutic protein preparations can be lyophilized and stored as sterile powders, preferably under vacuum, and then reconstituted in bacteriostatic water (containing for example, benzyl alcohol preservative) or in sterile water prior to injection.

Dosages and adminIstration protocols for the treatment of cancers using the foregoing methods will vary with the method and the target cancer, and will generally depend on a number of other factors appreciated In the art Cl(111.) Identificat Characterization and Use of Modulators of I Q9P1D Methods to Identify and Use Modulators In one embodiment, screening is performed to identify modulators that Induce or suppress a particular expression 00 profile, suppress or Induce specific pathways, preferably generating the associated phenotype thereby. In another 0 embodiment having Identified differentially expressed genes Important in a particular state; screens are performed to identify CK1 modulators that alter expression of individual genes, either increase or decrease. In another embodiment screening Is performed to identify modulators that alter a biological function of the expression product of a differentially expressed gene.

Again, having Identified the Importance of a gene In a particular state, screens are performed to Identify agents that bind andtor modulate the biological activity of the gene product In addition, screens are done for genes that are induced In response to a candidate agent After Identifying a modulator (one that suppresses a cancer expression pattern leading to a normal expression pattern, or a modulator of a cancer gene that leads to expression of the gene as in normal tissue) a screen is performed to Identify genes that are specifically modulated in response to the agent Comparing expression profiles between normal tissue and agent-treated cancer tissue reveals genes that are not expressed In normal tissue or cancer tissue, but are expressed in agent treated tissue, and vice versa. These agent-specific sequences are identified and used by methods described herein for cancer genes or proteins. In particular these sequences and the proteins they encode are used In marldng or Identifying agenttrated cells. In addition, antibodies are raised against the agent-Induced proteins and used to target novel therapeutics to the treated cancer tissue sample.

Modulator-related Identification and Screening Assas: Gene ENmession-related Assays Proteins, nucleic: acids, and antibodies of the Invention are used In screening assays. The cancer-associated proteins, antibodies, nucleic acids, modified proteins and cells containing these sequences are used In screening assays, such as evaluating the effect of drug candidates on a 'gene expression profile," expression profile of polypeptides or alteration of biological function. In one embodiment the expression profiles are used, preferably In conjunction with high throughput screening techniques to allow monitoring for expression proffle genes after treatment with a candidate agent Davis, GF, et al, J Bioll Screen 7:69 (2002); Zlokamik, et al., Science 279:84-8 (1998); Held, Genome Res 6:986- 94,1996).

The cancer proteins, antibodies, nucleic acids, modified proteins and cells containing the native or modified cancer proteins or genes are used in screening assays. That is, the present Invention comprises methods for screening for compositions which modulate the cancer phenotype or a physiological function of a cancer protein of the Invention. This is done on a gene Itself or by evaluating the effect of drug candidates on a "gene expression profile' or biological function. In one embodiment expression profiles are used, preferably in conjunction with high throughput screening techniques to allow 00 monitoring after treatment with a candidate agent, see Zlokamik, supra.

SA variety of assays are executed directed to the genes and proteins of the invention. Assays are run on an individual nucleic acid or protein level. That is, having identified a particular gene as up regulated in cancer, test compounds are screened for the ability to modulate gene expression or for binding to the cancer protein of the invention. 'Modulation' in Sthis context includes an increase or a decrease in gene expression. The preferred amount of modulation will depend on the Soriginal change of the gene expression In normal versus tissue undergoing cancer, with changes of at least 10%, preferably more preferably 100-300%, and in some embodiments 300-1000% or greater. Thus, if a gene exhibits a 4-fold increase in cancer tissue compared to normal tissue, a decrease of about four-fold is often desired; similarly, a C- decrease in cancer tissue compared to normal tissue a target value of a 10-fold increase in expression by the test compound Sis often desired. Modulators that exacerbate the type of gene expression seen in cancer are also useful, as an upregulated target in further analyses.

00 The amount of gene expression Is monitored using nucleic acid probes and the quantification of gene expression Slevels, or, alternatively, a gene product itself is monitored, through the use of antibodies to the cancer protein and C- standard immunoassays. Proteomics and separation techniques also allow for quantification of expression.

Expression Monitoring to Identify Comounds that Modify Gene Expression In one embodiment, gene expression monitoring, an expression profile, is monitored simultaneously for a number of entities. Such profiles will typically involve one or more of the genes of Figure 2. In this embodiment, cancer nudeic acid probes are attached to blochips to detect and quantify cancer sequences in a particular cell. Altematively, PCR can be used. Thus, a series, wells of a microtiter plate, can be used with dispensed primers in desired wells. A PCR reaction can then be performed and analyzed for each well.

Expression monitoring is performed to identify compounds that modify the expression of one or more cancerassodated sequences, a polynucleotide sequence set out in Figure 2. Generally, a test modulator is added to the cells prior to analysis. Moreover, screens are also provided to identify agents that modulate cancer, modulate cancer proteins of the invention, bind to a cancer protein of the invention, or interfere with the binding of a cancer protein of the invention and an antibody or other binding partner.

In one embodiment, high throughput screening methods involve providing a library containing a large number of potential therapeutic compounds (candidate compounds). Such "combinatorial chemical libraries' are then screened in one or more assays to identify those library members (particular chemical species or subclasses) that display a desired characteristic activity. The compounds thus identified can serve as conventional "lead compounds," as compounds for screening, or as therapeutics.

In certain embodiments, combinatorial libraries of potential modulators are screened for an ability to bind to a cancer polypeptide or to modulate activity. Conventionally, new chemical entities with useful properties are generated by identifying a chemical compound (called a "lead compound') with some desirable property or activity, inhibiting activity, creating variants of the lead compound, and evaluating the property and activity of those variant compounds. Often, high throughput screening (HTS) methods are employed for such an analysis.

As noted above, gene expression monitoring is conveniently used to test candidate modulators protein, nucleic acid or small molecule). After the candidate agent has been added and the cells allowed to incubate for a period, the sample containing a target sequence to be analyzed is, added to a biochip.

If required, the target sequence is prepared using known techniques. For example, a sample is treated to lyse the cells, using known lysis buffers, electroporation, etc., with purification and/or amplification such as PCR performed as appropriate. For example, an in vitro transcription with labels covalently attached to the nucleotides is performed. Generally, 00 the nucleic acids are labeled with biotin-FITC or PE, or with cy3 or SThe target sequence can be labeled with, a fluorescent, a chemiluminescent, a chemical, or a radioactive signal, to provide a means of detecting the target sequence's specific binding to a probe. The label also can be an enzyme, such as alkaline phosphatase or horseradish peroxidase, which when provided with an appropriate substrate produces a Sproduct that is detected. Altematively, the label is a labeled compound or small molecule, such as an enzyme inhibitor, that binds but is not catalyzed or altered by the enzyme. The label also can be a moiety or compound, such as, an epitope tag or biotin which specifically binds to streptavidin. For the example of biotin, the streptavidin is labeled as described above, thereby, providing a detectable signal for the bound target sequence. Unbound labeled streptavidin is typically removed prior Sto analysis.

O As will be appreciated by those in the art, these assays can be direct hybridization assays or can comprise "sandwich assays", which include the use of multiple probes, as is generally outlined in U.S. Patent Nos. 5, 681,702; 00 5,597,909; 5,545,730; 5,594,117; 5,591,584; 5,571,670; 5,580,731; 5,571,670; 5,591,584; 5,624,802; 5,635,352; 5,594,118; O 5,359,100; 5,124, 246; and 5,681,697. In this embodiment, in general, the target nucleic acid is prepared as outlined above, and then added to the biochip comprising a plurality of nucleic acid probes, under conditions that allow the formation of a hybridization complex.

A variety of hybridization conditions are used in the present invention, including high, moderate and low stringency conditions as outlined above. The assays are generally run under stringency conditions which allow formation of the label probe hybridization complex only in the presence of target Stringency can be controlled by altering a step parameter that is a thermodynamic variable, including, but not limited to, temperature, formamide concentration, salt concentration, chaotroplc salt concentration pH, organic solvent concentration, etc. These parameters may also be used to control non-specific binding, as is generally outlined in U.S. Patent No. 5,681,697. Thus, it can be desirable to perform certain steps at higher stringency conditions to reduce non-specific binding.

The reactions outlined herein can be accomplished in a variety of ways. Components of the reaction can be added simultaneously, or sequentially, in different orders, with preferred embodiments outlined below. In addition, the reaction may include a variety of other reagents. These include salts, buffers, neutral proteins, e.g. albumin, detergents, etc. which can be used to facilitate optimal hybridization and detection, and/or reduce nonspecific or background interactions. Reagents that otherwise improve the efficiency of the assay, such as protease Inhibitors, nudease inhibitors, anti-microbial agents, etc., may also be used as appropriate, depending on the sample preparation methods and purity of the target The assay data are analyzed to determine the expression levels of individual genes, and changes in expression levels as between states, forming a gene expression profile.

Biological Activity-related Assays The invention provides methods identify or screen for a compound that modulates the activity of a cancer-related gene or protein of the invention. The methods comprise adding a test compound, as defined above, to a cell comprising a cancer protein of the invention. The cells contain a recombinant nucleic acid that encodes a cancer protein of the invention.

In another embodiment, a library of candidate agents is tested on a plurality of cells.

In one aspect, the assays are evaluated in the presence or absence or previous or subsequent exposure of physiological signals, e.g. hormones, antibodies, peptides, antigens, cytokines, growth factors, action potentials, pharmacological agents including chemotherapeutics, radiation, cardnogenics, or other cells cell-cell contacts). In another example, the determinations are made at different stages of the cell cycle process. In this way, compounds that modulate genes or proteins of the invention are Identified. Compounds with pharmacological activity are able to enhance or <0 interfere with the activity of the cancer protein of the invention. Once identified, similar structures are evaluated to identify Scritical structural features of the compound.

In one embodiment, a method of modulating inhibiting) cancer cell division is provided; the method comprises administration of a cancer modulator. In another embodiment, a method of modulating inhibiting) cancer is Sprovided; the method comprises administration of a cancer modulator. In a further embodiment, methods of treating cells or individuals with cancer are provided; the method comprises administration of a cancer modulator.

In one embodiment, a method for modulating the status of a cell that expresses a gene of the invention is provided.

As used herein status comprises such art-accepted parameters such as growth, proliferation, survival, function, apoptosis, Ssenescence, location, enzymatic activity, signal transduction, etc. of a cell. In one embodiment, a cancer inhibitor is an Santibody as discussed above. In another embodiment, the cancer Inhibitor is an antisense molecule. A variety of cell growth, proliferation, and metastasis assays are known to those of skill in the art, as described herein.

00 0 Hih Throuahout Screening to Identify Modulators N The assays to identify suitable modulators are amenable to high throughput screening. Preferred assays thus detect enhancement or inhibition of cancer gene transcription, inhibition or enhancement of polypeptide expression, and inhibition or enhancement of polypeptide activity.

In one embodiment, modulators evaluated in high throughput screening methods are proteins, often naturally occurring proteins or fragments of naturally occurring proteins. Thus, cellular extracts containing proteins, or random or directed digests of proteinaceous cellular extracts, are used. In this way, libraries of proteins are made for screening in the methods of the invention. Particularly preferred in this embodiment are libraries of bacterial, fungal, viral, and mammalian proteins, with the latter being preferred, and human proteins being especially preferred. Particularly useful test compound will be directed to the class of proteins to which the target belongs, substrates for enzymes, or ligands and receptors.

Use of Soft Agar Growth and Colony Formation to Identify and Characterize Modulators Normal cells require a solid substrate to attach and grow. When cells are transformed, they lose this phenotype and grow detached from the substrate. For example, transformed cells can grow in stirred suspension culture or suspended in semi-solid media, such as semi-solid or soft agar. The transformed cells, when transfected with tumor suppressor genes, can regenerate normal phenotype and once again require a solid substrate to attach to and grow. Soft agar growth or colony formation in assays are used to identify modulators of cancer sequences, which when expressed in host cells, inhibit abnormal cellular proliferation and transformation. A modulator reduces or eliminates the host cells' ability to grow suspended in solid or semisolid media, such as agar.

Techniques for soft agar growth or colony formation in suspension assays are described in Freshney, Culture of Animal Cells a Manual of Basic Technique (3rd ed., 1994). See also, the methods section of Garkavtsev et al. (1996), supra.

Evaluation of Contact Inhibition and Growth Density imitation to Identify and Characterize Modulators Normal cells typically grow in a flat and organized pattern in cell culture until they touch other cells. When the cells touch one another, they are contact inhibited and stop growing. Transformed cells, however, are not contact Inhibited and continue to grow to high densities in disorganized foci. Thus, transformed cells grow to a higher saturation density than corresponding normal cells. This is detected morphologically by the formation of a disoriented monolayer of cells or cells in foci. Alternatively, labeling index with 3 H}-thymidine at saturation density is used to measure density limitation of growth, similarly an MTT or Alamar blue assay will reveal proliferation capacity of cells and the the ability of modulators to affect same. See Freshney (1994), supra. Transformed cells, when transfected with tumor suppressor genes, can regenerate a normal phenotype and become contact Inhibited and would grow to a lower density.

00 In this assay, labeling index with 3 H)-thymidine at saturation density is a preferred method of measuring density limitation of growth. Transformed host cells are transfected with a cancer-associated sequence and are grown for 24 hours at saturation density in non-limiting medium conditions. The percentage of cells labeling with 3 H)-thymidine is determined by Incorporated cpm.

)Contact Independent growth is used to Identify modulators of cancer sequences, which had led to abnormal u cellular proliferation and transformation. A modulator reduces or eliminates contact independent growth, and returns the cells to a normal phenotype.

Evaluation of Growth Factor or Serum Dependence to Identify and Characterize Modulators STransformed cells have lower serum dependence than their normal counterparts (see, Temin, J. Natl. Cancer SInst 37:167-175 (1966); Eagle et al., J. Exp. Med 131:836-879 (1970)); Freshney, supra. This is in part due to release of various growth factors by the transformed cells. The degree of growth factor or serum dependence of transformed host cells 0 can be compared with that of control. For example, growth factor or serum dependence of a cell is monitored in methods to 0identify and characterize compounds that modulate cancer-associated sequences of the invention.

Use of Tumor-specific Marker Levels to Identify and Characterize Modulators Tumor cells release an increased amount of certain factors (hereinafter 'tumor specific markers') than their normal counterparts. For example, plasminogen activator (PA) is released from human glioma at a higher level than from normal brain cells (see, Gullino, Angiogenesis, Tumor Vascularizatlon, and Potential Interference with Tumor Growth, in Biological Responses in Cancer, pp. 178-184 (Mihich 1985)). Similarly, Tumor Angiogenesis Factor (TAF) is released at a higher level in tumor cells than their normal counterparts. See, Folkman, Angiogenesis and Cancer, Sem Cancer Biol. (1992)), while bFGF is released from endothelial tumors (Ensoli, B et al).

Various techniques which measure the release of these factors are described in Freshney (1994), supra. Also, see, Unkless et al., J. Biol. Chem. 249:4295-4305 (1974); Strickland Beers, J. Biol. Chem. 251:5694-5702 (1976); Whur et al., Br. J. Cancer 42:305 312 (1980); Gullino, Angiogenesis, Tumor Vascularization, and Potential Interference with Tumor Growth, in Biological Responses In Cancer, pp. 178-184 (Mihich 1985); Freshney, Anticancer Res. 5:111-130 (1985).

For example, tumor specific marker levels are monitored in methods to identify and characterize compounds that modulate cancer-associated sequences of the invention.

Invasiveness into Matrigel to Identify and Characterize Modulators The degree of invasiveness into Matrigel or an extracellular matrix constituent can be used as an assay to identify and characterize compounds that modulate cancer associated sequences. Tumor cells exhibit a positive correlation between malignancy and invasiveness of cells into Matrigel or some other extracellular matrix constituent In this assay, tumorigenic cells are typically used as host cells. Expression of a tumor suppressor gene in these host cells would decrease invasiveness of the host cells. Techniques described in Cancer Res. 1999; 59:6010; Freshney (1994), supra, can be used.

Briefly, the level of invasion of host cells is measured by using filters coated with Matrigel or some other extracellular matrix constituent Penetration into the gel, or through to the distal side of the filter, is rated as Invasiveness, and rated histologically by number of cells and distance moved, or by prelabeling the cells with M1 and counting the radioactivity on the distal side of the filter or bottom of the dish. See, Freshney (1984), supra.

Evaluation of Tumor Growth In Vimo to Identify and Characterize Modulators Effects of cancer-associated sequences on cell growth are tested in transgenic or immune-suppressed organisms.

Transgenic organisms are prepared in a variety of art-accepted ways. For example, knock-out transgenic organisms, e.g., mammals such as mice, are made, in which a cancer gene is disrupted or In which a cancer gene Is inserted. Knock-out transgenic mice are made by insertion of a marker gene or other heterologous gene into the endogenous cancer gene site In 0 0 the mouse genome via homologous recombination. Such mice can also be made by substituting the endogenous cancer 0 gene with a mutated version of the cancer gene, or by mutating the endogenous cancer gene, by exposure to carcinogens.

To prepare transgenic chimeric animals, mice, a DNA construct is introduced Into the nuclei of embryonic Sstem cells. Cells containing the newly engineered genetic lesion are injected into a host mouse embryo, which is re- Simplanted into a recipient female. Some of these embryos develop into chimeric mice that possess germ cells some of which are derived from the mutant cell line. Therefore, by breeding the chimeric mice it is possible to obtain a new line of mice containing the introduced genetic lesion (see, Capecchi et al., Science 244:1288 (1989)). Chimeric mice can be derived according to US Patent 6,365,797, issued 2 April 2002; US Patent 6,107,540 issued 22 August 2000; Hogan et al., Manipulating the Mouse Embryo: A laboratory Manual, Cold Spring Harbor Laboratory (1988) and Teratocarcnomas and Embryonic Stem Cells: A Practical Approach, Robertson, ed., IRL Press, Washington, (1987).

00 Altematively, various immune-suppressed or Immune-deficient host animals can be used. For example, a Sgenetically athymic "nude" mouse (see, Glovanella et al., J. Natl. Cancer Inst 52:921 (1974)), a SCID mouse, a thymedomized mouse, or an irradiated mouse (see, Bradley et al., Br. J. Cancer 38:263 (1978); Selby et al., Br. J.

Cancer 41:52 (1980)) can be used as a host Transplantable tumor cells (typically about 106 cells) injected into isogenic hosts produce invasive tumors in a high proportion of cases, while normal cells of similar origin will not In hosts which developed invasive tumors, cells expressing cancer-associated sequences are injected subcutaneously or orthotopically.

Mice are then separated into groups, including control groups and treated experimental groups) e.g. treated with a modulator). After a suitable length of time, preferably 4-8 weeks, tumor growth is measured by volume or by its two largest dimensions, or weight) and compared to the control. Tumors that have statistically significant reduction (using, e.g., Student's T test) are said to have inhibited growth.

In Vitro Assays to Identify and Characterize Modulators Assays to identify compounds with modulating activity can be performed in vitro. For example, a cancer polypeptide is first contacted with a potential modulator and incubated for a suitable amount of time, from 0.5 to 48 hours. In one embodiment, the cancer polypeptide levels are determined in vitro by measuring the level of protein or mRNA.

The level of protein is measured using Immunoassays such as Westem blotting, ELISA and the like with an antibody that selectively binds to the cancer polypeptide or a fragment thereof. For measurement of mRNA, amplification, using PCR, LCR, or hybridization assays, e. Northem hybridization, RNAse protection, dot blotting, are preferred. The level of protein or mRNA is detected using directly or indirectly labeled detection agents, fluorescently or radioactively labeled nuclec adds, radioactively or enzymatically labeled antibodies, and the like, as described herein.

Alternatively, a reporter gene system can be devised using a cancer protein promoter operably linked to a reporter gene such as luciferase, green fluorescent protein, CAT, or P-gal. The reporter construct is typically transfected Into a cell.

After treatment with a potential modulator, the amount of reporter gene transcription, translation, or activity is measured according to standard techniques known to those of skill in the art (Davis GF, supra; Gonzalez, J. Negulescu, P. Curr.

Opin. Biotechnol. 1998: 9:624).

As outlined above, in vitro screens are done on individual genes and gene products. That Is, having Identified a particular differentially expressed gene as important in a particular state, screening of modulators of the expression of the gene or the gene product itself Is performed.

In one embodiment, screening for modulators of expression of specific gene(s) is performed. Typically, the expression of only one or a few genes is evaluated. In another embodiment, screens are designed to first find compounds 0 that bind to differentially expressed proteins. These compounds are then evaluated for the ability to modulate differentially Sexpressed activity. Moreover, once initial candidate compounds are identified, variants can be further screened to better evaluate structure activity relationships.

Binding Assays to Identify and Characterize Modulators In binding assays in accordance with the Invention, a purified or isolated gene product of the invention is generally used. For example, antibodies are generated to a protein of the invention, and immunoassays are run to determine the amount and/or location of protein. Alternatively, cells comprising the cancer proteins are used in the assays.

C Thus, the methods comprise combining a cancer protein of the invention and a candidate compound such as a Sligand, and determining the binding of the compound to the cancer protein of the invention. Preferred embodiments utilize the human cancer protein; animal models of human disease of can also be developed and used. Also, other analogous 00 mammalian proteins also can be used as appreciated by those of skill in the art Moreover, in some embodiments variant or Sderivative cancer proteins are used.

C= Generally, the cancer protein of the invention, or the ligand, is non-diffusibly bound to an Insoluble support. The support can, be one having isolated sample receiving areas (a microtiter plate, an array, etc.). The insoluble supports can be made of any composition to which the compositions can be bound, is readily separated from soluble material, and is otherwise compatible with the overall method of screening. The surface of such supports can be solid or porous and of any convenient shape.

Examples of suitable insoluble supports Include microtiter plates, arrays, membranes and beads. These are typically made of glass, plastic polystyrene), polysaccharide, nylon, nitrocellulose, or Teflon 1 etc. Microtiter plates and arrays are especially convenient because a large number of assays can be carried out simultaneously, using small amounts of reagents and samples. The particular manner of binding of the composition to the support is not crucial so long as it is compatible with the reagents and overall methods of the invention, maintains the activity of the composition and is nondiffusable. Preferred methods of binding Include the use of antibodies which do not sterically block either the ligand binding site or activation sequence when attaching the protein to the support, direct binding to "sticky" or ionic supports, chemical crosslinking, the synthesis of the protein or agent on the surface, etc. Following binding of the protein or ligand/binding agent to the support, excess unbound material is removed by washing. The sample receiving areas may then be blocked through incubation with bovine serum albumin (BSA), casein or other Innocuous protein or other moiety.

Once a cancer protein of the invention is bound to the support, and a test compound is added to the assay.

Altematively, the candidate binding agent is bound to the support and the cancer protein of the invention is then added.

Binding agents include specific antibodies, non-natural binding agents identified in screens of chemical libraries, peptide analogs, etc.

Of particular interest are assays to identify agents that have a low toxicity for human cells. A wide variety of assays can be used for this purpose, including proliferation assays, cAMP assays, labeled in vitro protein-protein binding assays, electrophoretic mobility shift assays, immunoassays for protein binding, functional assays (phosphorylation assays, etc.) and the like.

A determination of binding of the test compound (ligand, binding agent, modulator, etc.) to a cancer protein of the invention can be done in a number of ways. The test compound can be labeled, and binding determined directly, by attaching all or a portion of the cancer protein of the invention to a solid support, adding a labeled candidate compound a fluorescent label), washing off excess reagent, and determining whether the label Is present on the solid support. Various blocking and washing steps can be utilized as appropriate.

00 0 In certain embodiments, only one of the components is labeled, a protein of the invention or ligands labeled.

0 Alternatively, more than one component Is labeled with different labels, i112, for the proteins and a fluorophor for the compound. Proximity reagents, quenching or energy transfer reagents are also useful.

Comrpetitive Binding to Identiy and Characterize Modulators In one embodiment the binding of the 'test compound' is deternined by competitive binding assay with a 'competitor.' The competitor is a binding moiety that binds to the target molecule a cancer protein of the invention).

Competitors include compounds such as antbodies, peptides, binding partners, ligands, etc. Under certain circumstances, c=KI the competitive binding between the test compound and the competitor displaces the test compound. In one embodiment, the test compound is labeled. Either the test compound, the competitor, or both, Is added to the protein for a time sufficient to allow binding. Incubations are performed at a temperature that facilitates optimal activity, typically between four and 400C.

00 Incubation peuiods are typcaly optimized, to facilitate rapid high throughput screening; typically between zero and one hour will be sufficient. Excess reagent Is generally removed or washed away. The second component is then added, and ~K1 the presence or absence of the labeled component Is followed, to indicate binding.

In one embodiment the competitor Is added first followed by the test compound. Displacement of the competitor is an Indication that the test compound Is binding to the cancer protein and thus Is capable of binding to, and potentially modulating, the activity of the cancer protein. In this embodiment either component can be labeled. Thus, if the competitor is labeled, the presence of label In the post-test compound wash solution indicates displacement by the test compound. Alternatively, if the test compound is labeled, the presence of the label on the support indicates displacement In an alternative embodiment, the test compound is added first, with incubation and washing, followed by the competitor. The absence of binding by the competitor Indicates that the test compound binds to the cancer protein with higher affinity than the competitor. Thus, if the test compound Is labeled, the presence of the label on the support coupled with a lack of competitor binding, Indicates that the test compound binds to and thus potentially modulates the cancer protein of the invention.

Accordingly, the competitive binding methods comprise differential screening to Identity agents that are capable of modulating the activity of the cancer proteins of the invention. In this embodiment the methods comprise combining a cancer protein and a competitor in a first sample. A second sample comprises a test compound, the cancer protein, and a competitor. The binding of the competitor is determined for both samples, and a change, or difference in binding between the two samples Indicates the presence of an agent capable of binding to the cancer protein and potentially modulating its activity. That is, if the binding of the competitor is different in the second sample relative to the first sample, the agent Is capable of binding to the cancer protein.

Alternatively, differentiall screening Is used to identify drug candidates that bind to the native cancer protein, but cannot bind to modified cancer proteins. For example the structure of the cancer protein is modeled and used in rational drug design to synthesize agents that Interact with that site, agents which generally do not bind to site-modified proteins.

Moreover, such drug candidates that affect the activity of a native cancer protein are also Identified by screening drugs for the ability to either enhance or reduce the activity of such proteins.

Positive controls and negative controls can be used in the assays. Preferably control and test samples are performed In at least triplicate to obtain statistilly significant results. Incubation of all samples occurs for a time sufficient to allow for the binding of the agent to the protein. Following Incubation, samples are washed free of non-specifically bound material and the amount of bound, generally labeled agent determined. For example, where a radiolabel Is employed, the samples can be counted in a scintillation counter to determine the amount of bound compound.

00 A variety of other reagents can be included in the screening assays. These include reagents like salts, neutral 0proteins, e.g. albumin, detergents, etc. which are used to facilitate optimal protein-protein binding andlor reduce non-specific or background interactions. Also reagents that otherwise improve the efficiency of the assay, such as protease inhibitors, nudease Inhibitors, anti-microbial agents, etc., can be used. The mixture of components is added In an order that provides Sfor the requisite binding.

Use of Polynudeotides to Down-regulate or Inhibit a Protein of the Invention.

Polynucleotide modulators of cancer can be introduced into a cell containing the target nucleotide sequence by Sformation of a conjugate with a igand-binding molecule, as described in WO 91/04753. Suitable ligand-binding molecules Sinclude, but are not limited to, cell surface receptors, growth factors, other cytokines, or other ligands that bind to cell surface receptors. Preferably, conjugation of the Ilgand binding molecule does not substantially interfere with the ability of the ligand 0 binding molecule to bind to its corresponding molecule or receptor, or block entry of the sense or antisense oligonucleotide 0 or its conjugated version into the cell. Alternatively, a polynucleotide modulator of cancer can be introduced Into a cell C- containing the target nucleic acid sequence, by formation of a polynucleotide-lipid complex, as described in WO 90/10448. It is understood that the use of antisense molecules or knock out and knock in models may also be used in screening assays as discussed above, in addition to methods of treatment Inhibitory and Antisense Nucleotides In certain embodiments, the activity of a cancer-associated protein is down-regulated, or entirely inhibited, by the use of antisense polynudeotide or inhibitory small nuclear RNA (snRNA), a nucleic acid complementary to, and which can preferably hybridize specifically to, a coding mRNA nucleic acid sequence, a cancer protein of the invention, mRNA, or a subsequence thereof. Binding of the antisense polynucleotide to the mRNA reduces the translation and/or stability of the mRNA.

In the context of this invention, antisense polynudeotides can comprise naturally occurring nucleotides, or synthetic species formed from naturally occurring subunits or their close homologs. Antisense polynudeotides may also have altered sugar moieties or inter-sugar linkages. Exemplary among these are the phosphorothioate and other sulfur containing species which are known for use in the art Analogs are comprised by this invention so long as they function effectively to hybridize with nucleotides of the invention. See, Isis Pharmaceuticals, Carlsbad, CA; Sequitor, Inc., Natick, MA.

Such antisense polynudeotides can readily be synthesized using recombinant means, or can be synthesized in vitro. Equipment for such synthesis is sold by several vendors, Including Applied Biosystems. The preparation of other oligonudeotides such as phosphorothioates and alkylated derivatives is also well known to those of skill in the art Antisense molecules as used herein include antisense or sense oligonucleotides. Sense oligonucleotides can, be employed to block transcription by binding to the anti-sense strand. The antisense and sense oligonudeotide comprise a single stranded nucleic acd sequence (either RNA or DNA) capable of binding to target mRNA (sense) or DNA (antisense) sequences for cancer molecules. Antisense or sense oligonucleotides, according to the present invention, comprise a fragment generally at least about 12 nucleotides, preferably from about 12 to 30 nudeotides. The ability to derive an antisense or a sense oligonucleotide, based upon a cDNA sequence encoding a given protein is described in, Stein &Cohen (Cancer Res. 48:2659 (1988 and van der Krol et al. (BioTechniques 6:958 (1988)).

RIbozvmes In addition to antisense polynudeotides, ribozymes can be used to target and inhibit transcription of cancerassociated nudeotide sequences. A ribozyme is an RNA molecule that catalytically cleaves other RNA molecules. Different 00 0 kinds of ribozymes have been described, including group I ribozymes, hammerhead ribozymes, hairpin ribozymes, RNase P, Sand axhead nrbozymes (see, Castanotto et al., Adv. in Pharmacology 25: 289-317 (1994) for a general review of the properties of different ribozymes).

The general features of hairpin ribozymes are described, in Hampel et al., Nud. Adds Res. 18:299-304 (1990); European Patent Publication No. 0360257; U.S. Patent No. 5,254,678. Methods of preparing are well known to V those of skill in the art (see, WO 94/26877; Ojwang et al., Proc. Natl. Acad. Sci. USA 90:6340-6344 (1993); Yamada et al., Human Gene Therapy 1:39-45 (1994); Leavitt et al., Proc. Natl. Acad Sc. USA 92:699- 703 (1995); Leavitt et al., Human Gene Therapy 5:1151-120 (1994); and Yamada et al., Virology 205:121-126 (1994)).

SUse of Modulators in Phenotypic Screening In one embodiment, a test compound is administered to a population of cancer cells, which have an associated 00 cancer expression profile. By 'administration" or 'contacting" herein Is meant that the modulator is added to the cells in such Sa manner as to allow the modulator to act upon the cell, whether by uptake and intracellular action, or by action at the cell C< surface. In some embodiments, a nucleic acid encoding a proteinaceous agent a peptide) is put Into a viral construct such as an adenoviral or retroviral construct, and added to the cell, such that expression of the peptide agent is accomplished, PCT US97/01019. Regulatable gene therapy systems can also be used. Once the modulator has been administered to the cells, the cells are washed if desired and are allowed to incubate under preferably physiological conditions for some period. The cells are then harvested and a new gene expression profile is generated. Thus, e.g., cancer tissue is screened for agents that modulate, induce or suppress, the cancer phenotype. A change in at least one gene, preferably many, of the expression profile indicates that the agent has an effect on cancer activity. Similary, altering a biological function or a signaling pathway is indicative of modulator activity. By defining such a signature for the cancer phenotype, screens for new drugs that alter the phenotype are devised. With this approach, the drug target need not be known and need not be represented In the original gene/protein expression screening platform, nor does the level of transcript for the target protein need to change. The modulator inhibiting function will serve as a surrogate marker As outlined above, screens are done to assess genes or gene products. That is, having identified a particular differentially expressed gene as important in a particular state, screening of modulators of either the expression of the gene or the gene product itself is performed.

Use of Modulators to Affect Peptides of the Invention Measurements of cancer polypeptide activity, or of the cancer phenotype are performed using a variety of assays.

For example, the effects of modulators upon the function of a cancer polypeptide(s) are measured by examining parameters described above. A physiological change that affects activity is used to assess the influence of a test compound on the polypeptides of this invention. When the functional outcomes are determined using intact cells or animals, a variety of effects can be assesses such as, in the case of a cancer associated with solid tumors, tumor growth, tumor metastasis, neovascularization, hormone release, transcriptional changes to both known and uncharacterized genetic markers by Northern blots), changes in cell metabolism such as cell growth or pH changes, and changes in Intracellular second messengers such as cGNIP.

Methods of Identifving Characterizing Cancer-associated Sequences Expression of various gene sequences is correlated with cancer. Accordingly, disorders based on mutant or variant cancer genes are determined. In one embodiment, the invention provides methods for identifying cells containing 00 variant cancer genes, determining the presence of, all or part the sequence of at least one endogenous cancer gene in a cell. This is accomplished using any number of sequencing techniques. The invention comprises methods of Identifying the cancer genotype of an individual, determining all or part of the sequence of at least one gene of the Invention in the Individual. This is generally done in at least one tissue of the individual, a tissue set forth in Table 1, and may include the evaluation of a number of tissues or different samples of the same tissue. The method may Include comparing the sequence of the sequenced gene to a known cancer gene, a wild-type gene to determine the presence of family members, homologies, mutations or variants. The sequence of all or part of the gene can then be compared to the sequence of a known cancer gene to determine If any differences exist This is done using any number of known homology programs, such as BLAST, Bestfit, etc. The presence of a difference in the sequence between the cancer gene of the patient and the known cancer gene correlates with a disease state or a propensity for a disease state, as outlined herein.

In a preferred embodiment, the cancer genes are used as probes to determine the number of copies of the cancer OC) gene In the genomne. The cancer genes are used as probes to determine the chromosomal localization of the cancer genes.

information such as chromosomal localization finds use In providing a diagnosis or prognosis In particular when chromosomal abnormalities such as translocatoris, and the like are Identified In the cancer gene locus.

XIV.) )Wltslicles of Manufacture For use in the laboratory, prognostic prophylactic, diagnostic and therapeutic applications described herein, kits are within the scope of the invention. Such kits can comprise a carrier, package, or container that is compartmnentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used In the method, along with a label or insert comprising Instructions for use, such as a use described herein. For example, the container(s) can comprise a probe that is or can be detectably labeled. Such probe can be an antibody or polyriucleotide specific for a protein or a gene or message of the invention, respectively. Where the method utilizes nucleic acid hybridization to detect the target nucleic acid, the kit can also have containers containing nucleotide(s) for amplification of the target nucleic acid sequence. Kits can comprise a container comprising a reporter, such as a biotinbinding protein, such as avidin or streptavIdin, bound to a reporter mnolecule, such as an enzymatic, fluorescent or radioisotope label; such a reporter can be used With, a nucleic acid or antibody. The kit can Indlude all or part of the amino acid sequences In Figure 2 or Figure 3 or analogs thereof, or a nucleic: acid molecule that encodes such amino acid sequences.

The kit of the invention will typically comprise the container described above and one or more other containers associated therewith that comprise materials desirable from a comm~ercial and user standpoint including buffers, diluents, filters, needles, syrnges; carrier, package, container, vial and/or tube labels listing contents and/or lnstnuclions for use, and package Inserts with Instructions for use.

A label can be present on or with the container to Indicate that the composition is used for aspecifictherapy or nontherapeutic application, such as a prognostic, prophylactic, diagnostic or laboratory application, and can also indicate directions for either ivio or i m use, such as those desc1,edherein. Drections and or other information can also beincluded on an insert(s) or label(s) whichIs Included with or onthe kit The label can be onor assocated with tMecontainer. A label acan be on a container when letters, numbers or other characters formIng the label are molded or etched into the container itself, a label can be associated with a container when it is present within a receptacle or carrier that also holds the container, as a package Insert The label can Indicate that the composition is used for diagnosing, treating, prophylaxing or prognosing a condition, such as a neoplasia of a tissue set fort in Table 1.

The terms Vr and 'article of manufacture' can be used as synonyms.

00 In another embodiment of the invention, an article(s) of manufacture containing compositions, such as amino acid sequence(s), small molecule(s), nucleic acid sequence(s), and/or antibody(s), materials useful for the diagnosis, prognosis, prophylaxis and/or treatment of neoplasias of tissues such as those set forth in Table I is provided. The article of q) manufacture typically comprises at least one container and at least one label. Suitable containers include, for example, Sbottles, vials, syringes, and test tubes. The containers can be formed from a variety of materials such as glass, metal or plastic. The container can hold amino acid sequence(s), small molecule(s), nudeic acid sequence(s), cell population(s) and/or antibody(s). In one embodiment the container holds a polynudeotide for use in examining the mRNA expression profile of a cell, together with reagents used for this purpose. In another embodiment a container comprises an antibody, binding fragment thereof or specific binding protein for use in evaluating protein expression of109P1D4 in cells and tissues, Sor for relevant laboratory, prognostic, diagnostic, prophylactic and therapeutic purposes; Indications and/or directions for such uses can be included on or with such container, as can reagents and other compositions or tools used for these 00 purposes. In another embodiment, a container comprises materials for eliciting a cellular or humoral immune response, 0 together with associated indications and/or directions. In another embodiment, a container comprises materials for adoptive Simmunotherapy, such as cytotoxic T cells (CTL) or helper T cells (HTL), together with associated indications and/or directions; reagents and other compositions or tools used for such purpose can also be Included.

The container can alternatively hold a composition that is effective for treating, diagnosis, prognosing or prophylaxing a condition and can have a sterile access port (for example the container can be an Intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). The active agents in the composition can be an antibody capable of specifically binding 109PiD4 and modulating the function of 109P1D4.

The artide of manufacture can further comprise a second container comprising a pharmaceutically-acceptable buffer, such as phosphate-buffered saline, Ringer's solution and/or dextrose solution. It can further include other materials desirable from a commercial and user standpoint including other buffers, diluents, filters, stirrers, needles, syringes, and/or package inserts with indications and/or instructions for use.

EXAMPLES:

Various aspects of the invention are further described and illustrated by way of the several examples that follow, none of which is intended to limit the scope of the invention.

Example 1: SSH-Generated Isolation of cDNA Fraament of the 109P1D4 Gene To Isolate genes that are over-expressed in prostate cancer we used the Suppression Subtractive Hybridization (SSH) procedure using cDNA derived from prostate cancer tissues. The 109Pi D4 SSH cDNA sequence was from an experiment where cONA derived from LNCaP cells that was androgen-deprived (by growing in the presence of charcoal-stripped serum) was subtracted from cDNA derived from LNCaP cells that were stimulated with mibolerone for 9 hours.

Materials and Methods Human Tissues: The patient cancer and normal tissues were purchased from different sources such as the NDRI (Philadelphia, PA).

mRNA for some normal tissues were purchased from different companies such as Clontech, Palo Alto, CA.

RNA Isolation: Tissues were homogenized in Trizol reagent (Ufe Technologies, Gibco BRL) using 10 ml/ g tissue to isolate total RNA.

Poly A RNA was purified from total RNA using Qiagen's Oligotex mRNA Mini and Midi kits. Total and mRNA were quantified by spectrophotometric analysis 260/280 nm) and analyzed by gel electrophoresis.

00 Oligonuceotides: The following HPLC purified oligonucleotides were used.

DPNCDN (cDNA synthesis primer): 5TTTTGATCAAGCTT 3 o3' (SEQ ID NO: 44) V~ Adaptor 1: 5'CTAATACGACTCACTATAGGGCTCGAGCGGCCGCCCGGGCAG3' (SEQ ID NO: (SEQ ID NO: 46) Adaptor2: S5'GTAATACGACTCACTATAGGGCAGCGTGGTCGCGGCCGAG3' (SEQ ID NO: 47) (SEQ ID NO: 48) 0 PCR primer 1: 00 05'CTAATACGACTCACTATAGGGC3' (SEQ ID NO: 49) SNested primer (NP)1: 5'TCGAGCGGCCGCCCGGGCAGGA3' (SEQ ID NO: Nested primer (NP)2: 5'AGCGTGGTCGCGGCCGAGGA3' (SEQ ID NO: 51) Suppression Subtractive Hybridization: Suppression Subtractive Hybridization (SSH) was used to identify cDNAs corresponding to genes that may be differentially expressed in prostate cancer. The SSH reaction utilized cDNA from LNCaP prostate cancer cells.

The 109P1D4 SSH sequence was derived from cDNA subtraction of LNCaP stimulated with mibolerone minus LNCaP in the absence of androgen. The SSH DNA sequence (Figure 1) was identified.

The cDNA derived from androgen-deprived LNCaP cells was used as the source of the "driver' cDNA, while the cDNA from androgen-stimulated LNCaP cells was used as the source of the tester" cDNA. Double stranded cONAs corresponding to tester and driver cDNAs were synthesized from 2 pg of poly(A) RNA isolated from the relevant xenograft tissue, as described above, using CLONTECH's PCR-Select cDNA Subtraction Kit and 1 pg of oligonuceotide DPNCDN as primer. First- and secondstrand synthesis were carried out as described in the Kit's user manual protocol (CLONTECH Protocol No. PT1117-1, Catalog No.

K1804-1). The resulting cDNA was digested with Dpn II for 3 hrs at 370C. Digested cDNA was extracted with phenol/chloroform and ethanol precipitated.

Tester cDNA was generated by diluting 1 p1 of Dpn II digested cDNA from the relevant tissue source (see above) (400 ng) in 5 pi of water. The diluted cDNA (2 pI, 160 ng) was then ligated to 2 pl of Adaptor 1 and Adaptor 2 (10 pM), in separate ligation reactions, in a total volume of 10 p at 16C overnight, using 400 pl ofT4 DNA ligase (CLONTECH). Ugation was terminated with 1 pl of 0.2 M EDTA and heating at 72°C for 5 min.

The first hybridization was performed by adding 1.5 p (600 ng) of driver cDNA to each of two tubes containing 1.5 pi ng) Adaptor 1- and Adaptor 2- ligated tester cDNA. In a final volume of 4 pJ, the samples were overlaid with mineral oil, denatured in an MJ Research thermal cycler at 980C for 1.5 minutes, and then were allowed to hybridize for 8 hrs at 680C. The two hybridizations were then mixed together with an additional 1 pi of fresh denatured driver cDNA and were allowed to hybridize overnight at 6800. The second hybridization was then diluted in 200 pd of 20 mM Hepes, pH 8.3, 50 mM NaCI, 0.2 mM EDTA, heated at 70oC for 7 min. and stored at -200C.

PCR Amdlfication. Cloning and Seauencing of Gene Fraaments Generated from SSH: 83 00 To amplify gene fragments resulting from SSH reactions, two PCR amplifications were performed. In the primary PCR 0 reaction 1 pl of the diluted final hybridization mix was added to 1 pl of PCR primer 1 (10 pM), 0.5 pl dNTP mix (10 pM), 2.5 pl x reaction buffer (CLONTECH) and 0.5 p 50 x Advantage cDNA polymerase Mix (CLONTECH) in a final volume of 25 pi. PCR 1 was conducted using the following conditions: 75oC for 5 min., 940C for 25 sec., then 27 cycles of 94oC for 10 sec, 66°C for 30 sec, 72oC for 1.5 min. Five separate primary PCR reactions were performed for each experiment The products were pooled and diluted 1:10 with water. For the secondary PCR reaction, 1 pl from the pooled and diluted primary PCR reaction was added to the same reaction mix as used for PCR 1, except that primers NP1 and NP2 (10 pM) were used instead of PCR primer 1. PCR 2 was performed using 10-12 cycles of 94cC for 10 sec, 680C for 30 sec, and 72oC for 1.5 minutes. The PCR products were analyzed C using 2% agarose gel electrophoresis.

S The PCR products were inserted Into pCR2.1 using the T/A vector cloning kit (Invitrogen). Transformed E. coil were Ssubjected to blue/white and ampicillin selection. White colonies were picked and arrayed into 96 well plates and were grown in 00 liquid culture overnight To identify Inserts, PCR amplification was performed on 1 pl of bacterial culture using the conditions of PCR1 and NP1 and NP2 as primers. PCR products were analyzed using 2% agarose gel electrophoresis.

c Bacterial clones were stored in 20% glycerol in a 96 well format Plasmid DNA was prepared, sequenced, and subjected to nucleic add homology searches of the GenBank, dBest, and NCI-CGAP databases.

RT-PCR Expression Analysis: First strand cDNAs can be generated from 1 pg of mRNA with oligo (dT)12-18 priming using the Gibco-BRL Superscript Preamplification system. The manufacturer's protocol was used which included an incubation for 50 min at 42°C with reverse transcriptase followed by RNAse H treatment at 37oC for 20 min. After completing the reaction, the volume can be increased to 200 pl with water prior to normalization. First strand cDNAs from 16 different normal human lissues can be obtained from Clontech.

Normalization of the first strand cDNAs from multiple tissues was performed by using the primers 5'ATATCGCCGCGCTCGTCGTCGACAA3' (SEQ ID NO: 52) and 5'AGCCACACGCAGCTCATTGTAGAAGG 3' (SEQ ID NO: 53) to amplify P-actin. First strand cDNAs (5 pi) were amplified in a total volume of 50 p; containing 0.4 pM primers, 0.2 pM each dNTPs, 1X PCR buffer (Clontech, 10 mM Tris-HCL, 1.5 mM MgCI2, 50 mM KCI, pH8.3) and 1X Klentaq DNA polymerase (Clontech). Five pi of the PCR reaction can be removed at 18, 20, and 22 cycles and used for agarose gel electrophoresis. PCR was performed using an MJ Research thermal cycler under the following conditions: Initial denaturation can be at 94C for 15 sec, followed by a 18, 20, and 22 cycles of 94C for 15, 650C for 2 min, 720C for 5 sec. A final extension at 72°C was carred out for 2 min. After agarose gel electrophoresis, the band Intensities of the 283 base pair p-actin bands from multiple tissues were compared by visual inspection. Dilution factors for the first strand cDNAs were calculated to result in equal p-actin band intensities in all tissues after 22 cycles of PCR. Three rounds of normalization can be required to achieve equal band intensities in all tissues after 22 cycles of PCR.

To determine expression levels of the 109P1 D4 gene, 5 pi of normalized first strand cDNA were analyzed by PCR using 26, and 30 cycles of amplification. Semi-quantitative expression analysis can be achieved by comparing the PCR products at cycle numbers that give light band intensities. The primers used for RT-PCR were designed using the 109P1D4 SSH sequence and are listed below: 109P1D4.1 TGGTCTTTCAGGTAATTGCTGTTG 3' (SEQ ID NO: 54) 109P1D4.2 CTCCATCAATGTTATGTTGCCTGT 3' (SEQ ID NO: 00 0 A typical RT-PCR expression analysis is shown in Figure Example 2: Isolation of Full Length 109PiD4 encoding DNA The 109P1 D4 SSH sequence of 192 bp (Figure 1) exhibited homology to protocadherin 11 (PCDH11), a cell adhesion molecule related to the calcium dependent cadherins. The human cDNA sequence encodes a 1021 amino acid protein with an Nterminal leader sequence and a transmembrane domain. 109P1D4 v.1 of 4603bp was cloned from human prostate cancer xenograft LAPC-9AD cDNA library, revealing an ORF of 1021 amino acids (Figure 2 and Figure Other variants (Transcript and SNP) of 109P1D4 were also identified and these are listed sequentially in Figure 2 and Figure 3.

(1 Example 3: Chromosomal Mapping of 109P1D4 Chromosomal localization can implicate genes in disease pathogenesis. Several chromosome mapping approaches are 00 available including fluorescent in situ hybridization (FISH), human/hamster radiation hybrid (RH) panels (Walter et al., 1994; SNature Genetics 7:22; Research Genetics, Huntsville Al), human-rodent somatic cell hybrid panels such as is available from the Cl Coriel Institute (Camden, New Jersey), and genomic viewers utilizing BLAST homologies to sequenced and mapped genomic clones (NCBI, Bethesda, Maryland).

109P1D4 maps to chromosome Xq21.3 using 109P1D4 sequence and the NCBI BLAST tool: located on the World Wide Web at (.ncbl.nlm.nih.gov/genome/seq/page.cgi?F=HsBlasthtml&&ORG=Hs). 109P1D4 was also Identified on chromosome Yp11.2, a region of 99% identity to Xq21.

Example 4: Expression Analysis of 109P1D4 in Normal Tissues and Patient Specimens Expression analysis by RT-PCR and Northern analysis demonstrated that normal tissue expression of a gene of Figure 2 is restricted predominantly to the tissues set forth in Table I.

Therapeutic applications for a gene of Figure 2 include use as a small molecule therapy and/or a vaccine (T cell or antibody) target Diagnostic applications for a gene of Figure 2 indude use as a diagnostic marker for local and/or metastasized disease. The restricted expression of a gene of Figure 2 in normal tissues makes it useful as a tumor target for diagnosis and therapy. Expression analysis of a gene of Figure 2 provides information useful for predicting susceptibility to advanced stage disease, rate of progression, and/or tumor aggressiveness. Expression status of a gene of Figure 2 in patient samples, tissue arrays and/or cell lines may be analyzed by: immunohistochemical analysis; (ii) In situ hybridization; (iii) RT-PCR analysis on laser capture micro-dissected samples; (iv) Western blot analysis; and Northern analysis.

RT-PCR analysis and Northern blotting were used to evaluate gene expression In a selection of normal and cancerous urological tissues. The results are summarized in Figures 15-19.

Figure 14 shows expression of 109P1D4 in lymphoma cancer patient specimens. RNA was extracted from peripheral blood lymphocytes, cord blood isolated from normal individuals, and from lymphoma patient cancer specimens.

Northern blots with 10 g of total RNA were probed with the 109P1D4 sequence. Size standards in kilobases are on the side. Results show expression of 109P1 D4 in lymphoma patient specimens but not in the normal blood cells tested.

Figure 15 shows expression of 109P1D4 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool, and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 109P1 D4, was performed at 30 cycles of amplification. Results show strong expression of 109P1 D4 in all cancer pools 00 tested. Very low expression was detected in the vital pools.

Figure 16 shows expression of 109P1D4 in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 pg of mRNAJane, were probed with the 109P1 D4 SSH fragment Size standards in kilobases (kb) are indicated on the side. Results show expression of approximately 10 kb 109P1 D4 transcript in ovary. Weak expression was also detected In placenta and brain, but not in the other normal tissues tested.

Figure 17 shows expression of 109P1 D4 in human cancer cell lines. RNA was extracted from a number of human prostate and bone cancer cell lines. Northern blots with 10 pg of total RNAlane were probed with the 109P1D4 SSH fragment Size standards in kilobases (kb) are indicated on the side. Results show expression of 109P1D4 in LAPC-9AD, LAPC-9AI, LNCaP prostate cancer cell lines, and in the bone cancer cell lines, SK-ES-1 and RD-ES.

Extensive expression of 109P1 D4 In normal tissues is shown in Figure 18A A cONA dot blot containing 76 different samples from human tissues was analyzed using a 109P1 D4 SSH probe. Expression was only detected In multiple areas of the brain, placenta, ovary, and fetal brain, amongst all tissues tested.

Figure 18B shows expression of 109P1D4 in patient cancer specimens. Expression of 109P1D4 was assayed in a panel of human cancers and their respective matched normal tissues on RNA dot blots. Upregulated expression of 109PID4 In tumors compared to normal tissues was observed In uterus, lung and stomach. The expression detected in normal adjacent tissues (isolated from diseased tissues) but not In normal tissues (isolated from healthy donors) may indicate that these tissues are not fully normal and that 109P1D4 may be expressed in early stage tumors.

Figure 19 shows 109P1D4 expression in lung cancer patient specimens. RNA was extracted from normal lung, prostate cancer xenograft LAPC-9AD, bone cancer cell line RD-ES, and lung cancer patient tumors. Northern blots with pg of total RNA were probed with 109P1D4. Size standards in klobases are on the side. Results show strong expression of 109PID4 in lung tumor tissues as well as the RD-ES cell line, but not in norma lung.

The restricted expression of 109P1D4 in normal tissues and the expression detected In cancer patient specimens suggest that 109P1 D4 is a potential therapeutic target and a diagnostic marker for human cancers.

Example 5: Splice Vadants of 1 09P1D4 Transcript variants are variants of mature mRNA from the same gene which arise by alternative transcription or alternative splicing. Alternative transcripts are transcripts from the same gene but start transcription at different points.

Splice variants are mRNA variants spliced differently from the same transcript In eukaryotes, when a multi.exon gene is transcribed from genomic DNA, the initial RNA is spliced to produce functional mRNA, which has only exons and is used for translation into an amino acid sequence. Accordingly, a given gene can have zero to many alternative transcripts and each transcript can have zero to many splice variants. Each transcript variant has a unique exon makeup, and can have different coding and/or non-coding or 3' end) portions, from the original transcript Transcript variants can code for similar or different proteins with the same or a similar function or can encode proteins with different functions, and can be expressed in the same tissue at the same time, or In different tissues at the same time, or In the same tissue at different times, or In different tissues at different times. Proteins encoded by transcript variants can have similar or different cellular or extracellular localizations, secreted versus intracelular.

Transcript variants are Identified by a variety of art-accepted methods. For example, alternative transcripts and splice variants are identified by full-length doning experiment or by use of full-length transcript and EST sequences. First all human ESTs were grouped into dusters which show direct or Indirect identity with each other. Second, ESTs in the same duster were further grouped Into sub-clusters and assembled into a consensus sequence. The original gene sequence Is compared to the consensus sequence(s) or other full-length sequences. Each consensus sequence Is a potential splice 00 variant for that gene. Even when a varant is identified that is not a full-length clone, that portion of the variant is very useful

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for antigen generation and for further cloning of the full-length splice variant using techniques known In the art.

c Moreover, computer programs are available in the art that identify transcript variants based on genomic a, sequences. Genomic-based transcript variant identification programs include FgenesH Salamov and V. Solovyev, 'Ab Sinitio gene finding in Drosophila genomic DNA," Genome Research. 2000 April;10(4):516-22); Grail (URL V compblo.oml.gov/Grail-bin/EmptyGrailForm) and GenScan (URL genes.mitedulGENSCAN.html). For a general discussion of splice variant identification protocols see., Southan, A genomic perspective on human proteases, FEBS Lett.

2001 Jun 8; 498(2-3):214-8; de Souza, et al, Identification of human chromosome 22 transcribed sequences with ORF Sexpressed sequence tags, Proc. Natl Acad Sc U S A. 2000 Nov 7; 97(23):12690-3.

STo further confirm the parameters of a transcript variant, a variety of techniques are available in the art, such as full-length cloning, proteomic validation, PCR-based validation, and 5' RACE validation, etc. (see Proteomic Validation: 0 Brennan, et al., Albumin banks peninsula: a new termination variant characterized by electrospray mass spectrometry, SBlochem Biophys Acta. 1999 Aug 17;1433(1-2):321-6; Ferranti P, et al., Differential splicing of pre-messenger RNA produces Smultiple forms of mature caprine alpha(sl)-caseln, Eur J Biochem. 1997 Oct 1;249(1):1-7. For PCR-based Validation: Wellmann S, et al, Specific reverse transcription-PCR quantification of vascular endothelial growth factor (VEGF) splice variants by LightCycler technology, Clin Chem. 2001 Apr;47(4):654-60; Jia, et al., Discovery of new human betadefensins using a genomics-based approach, Gene. 2001 Jan 24; 263(1-2):211-8. For PCR-based and 5' RACE Validation: Brigle, et Organization of the murine reduced folate carrier gene and identification of variant splice forms, Biochem Biophys Acta. 1997 Aug 7; 1353(2): 191-8).

It is known in the art that genomic regions are modulated in cancers. When the genomic region to which a gene maps is modulated in a particular cancer, the alternative transcripts or splice variants of the gene are modulated as well.

Disclosed herein is that 109P1 D4 has a particular expression profile related to cancer. Alternative transcripts and splice variants of 109P1D4 may also be involved in cancers in the same or different tissues, thus serving as tumor-associated markers/antigens.

Using the full-length gene and EST sequences, 8 transcript variants were identified, designated as 109P1D4 v.2, v.3, v.4, v.5, v.6, v.7, v.8 and v.9. The boundaries of the exon In the original transcript 109P1D4 v.1, were shown in Table LI.

Compared with 109P1D4 v.1, transcript variant 109P1D4 v.3 has spliced out 2069-2395 from variant 109P1D4 v.1, as shown in Figure 12. Variant 109P1D4 v.4 spliced out 1162-2096 of variant 109P1D4 v.1. Variant 109P1D4 v.5 added one exon to the 5' and extended 2 bp to the 5' end and 288 bp to the 3' end of variant 109P1D4 v.1. Theoretically, each different combination of exons in spatial order, e.g. exon 1 of v.5 and exons 1 and 2 of v.3 or v.4, is a potential splice variant Tables LII through LV are set forth on a variant-by-variant basis. Tables Lll(a)-(h) show nudeotide sequence of the transcript variants. Tables Lll(a)-(h) show the alignment of the transcript variants with nucleic acd sequence of 109P1D4 v.1. Tables LIV(a)-(h) lay out amino add translation of the transcript variants for the identified reading frame orientation. Tables LV(a)-h) displays alignments of the amino acid sequence encoded by the splice variants with that of 109P1D4 v.1.

Example 6: Single Nucleotide Polymorphlsms of 109P1D4 A Single Nudeotide Polymorphism (SNP) is a single base pair variation in a nucleotide sequence at a specific location. At any given point of the genome, there are four possible nucleotide base pairs: A/T, CIG, GIC and TIA. Genotype refers to the specific base pair sequence of one or more locations in the genome of an Individual. Haplotype refers to the base pair sequence of more than one location on the same DNA molecule (or the same chromosome in higher organisms),

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0 0 often in the context of one gene or in the context of several tightly linked genes. SNP that occurs on a cDNA is called cSNP.

SThis cSNP may change amino acids of the protein encoded by the gene and thus change the functions of the protein. Some

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SNP cause inherited diseases; others contribute to quantitative variations in phenotype and reactions to environmental factors including diet and drugs among individuals. Therefore, SNP andlor combinations of alleles (called haplotypes) have Smany applications, including diagnosis of inherited diseases, determination of drug reactions and dosage, identification of n genes responsible for diseases, and analysis of the genetic relationship between individuals Nowotny, J. M. Kwon and A.

M. Goate, SNP analysis to dissect human traits," Curr. Opin. Neurobiol. 2001 Oct; 11(5):637-641; M. Pirmohamed and B.

K. Park, "Genetic susceptibility to adverse drug reactions,' Trends Pharmacol. Sci. 2001 Jun; 22(6):298-305; J. H. Riley, C.

SJ. Allan, E. Lal and A. Roses, "The use of single nudeotide polymorphisms in the isolation of common disease genes," SPharmacogenomics. 2000 Feb; 1(1):39-47; R. Judson, J. C. Stephens and A. Windemuth, "The predictive power of haplotypes In clinical response," Pharmacogenomics. 2000 Feb; 1(1):15-26).

c SNP are identified by a variety of art-accepted methods Bean, "The promising voyage of SNP target discovery," Am. Clin. Lab. 2001 Oct-Nov; 20(9):18-20; K. M. Weiss, "In search of human variation," Genome Res. 1998 Jul; 8(7):691- 1 697; M. M. She, "Enabling large-scale pharmacogenetic studies by high-throughput mutation detection and genotyping technologies," Clin. Chem. 2001 Feb; 47(2):164-172). For example, SNP can be identified by sequencing DNA fragments that show polymorphism by gel-based methods such as restriction fragment length polymorphism (RFLP) and denaturing gradient gel electrophoresis (DGGE). They can also be discovered by direct sequencing of DNA samples pooled from different individuals or by comparing sequences from different DNA samples. With the rapid accumulation of sequence data in public and private databases, one can discover SNP by comparing sequences using computer programs Gu, L. Hillier and P. Y. Kwok, "Single nudeotide polymorphism hunting in cyberspace,' Hum. Mutat 1998; 12(4):221-225). SNP can be verified and genotype or haplotype of an individual can be determined by a variety of methods Including direct sequencing and high throughput microarrays Y. Kwok, "Methods for genotyping single nucleotide polymorphisms," Annu. Rev.

Genomics Hum. Genet. 2001; 2:235-258; M. Kokoris, K. Dix, K. Moynihan, J. Mathis, B. Erwin, P. Grass, B. Hines and A.

Duesterhoeft, "High-throughput SNP genotyping with the Masscode system," Mol. Diagn. 2000 Dec; 5(4):329-340).

Using the methods described above, SNP were identified in the original transcript 109P404 v.1, and its variants (see Figure 2J and Figure 2K). These alleles of the SNP, though shown separately here, can occur in different combinations (haplotypes) and in any one of the transcript variants (such as 109P4D4 v.4 or v.5) that contains the site of the SNP.

Transcript variants v.4 and v.5 contained those SNP in the exons shared with variant v.3, and transcript variant v.9 contained all the SNP occurred in variant v.6 (see Figure Example 7: Production of Recombinant 109P1D4 In Prokaryotic Systems To express recombinant 109P1D4 and 109P1D4 variants in prokaryotic cells, the full or partial length 109P1D4.

and 109P1D4 variant cDNA sequences are cloned into any one of a variety of expression vectors known in the art One or more of the following regions of 109P1D4 variants are expressed: the full length sequence presented in Figures 2 and 3, or any 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more contiguous amino acids from 109P1D4, variants, or analogs thereof.

A. In vitro transcription and translation constructs: pCRII: To generate 109P1D4 sense and anti-sense RNA probes for RNA in situ investigations, pCRII constructs (Invitrogen, Carlsbad CA) are generated encoding either all or fragments of the 109P1 D4 cDNA. The pCRII vector has Sp6 and T7 promoters flanking the insert to drive the transcription of 109P1D4 RNA for use as probes in RNA in situ hybridization experiments. These probes are used to analyze the cell and tissue expression of 109P1D4 at the RNA level. Transcribed 0 109P1D4 RNA representing the cDNA amino acid coding region of the 109P1D4 gene is used in in vitro translation systems such as the TnTrM Coupled Reticulolysate System (Promega, Corp., Madison, WI) to synthesize 109P1D4 protein.

B. Bacterial Constructs: 3) pGEX Constructs: To generate recombinant 109P1D4 proteins in bacteria that are fused to the Glutathione Stransferase (GST) protein, all or parts of the 109P1D4 cDNA protein coding sequence are cloned into the pGEX family of SGST-fusion vectors (Amersham Pharmacia Biotech, Piscataway, NJ). These constructs allow controlled expression of recombinant 109P1D4 protein sequences with GST fused at the amino-terminus and a six histidine epitope (6X His) at the carboxyl-terminus. The GST and 6X His tags permit purification of the recombinant fusion protein from induced bacteria with C1 the appropriate affinity matrix and allow recognition of the fusion protein with anti-GST and anti-His antibodies. The 6X His tag is generated by adding 6 histidine codons to the cloning primer at the 3' end, of the open reading frame (ORF). A proteolytic cleavage site, such as the PreSdsslon T recognition site in pGEX-6P-1, may be employed such that it permits 00 cleavage of the GST tag from 109P1D4-related protein. The ampicillin resistance gene and pBR322 origin permits selection Sand maintenance of the pGEX plasmids in E. coli.

p DMAL Constructs: To generate, in bacteria, recombinant 109P1D4 proteins that are fused to maltose-binding protein (MBP), all or parts of the 109P1D4 cONA protein coding sequence are fused to the MBP gene by cloning into the pMAL-c2X and pMAL-p2X vectors (New England Biolabs, Beverty, MA). These constructs allow controlled expression of recombinant 109P1D4 protein sequences with MBP fused at the amino-terminus and a 6X His epitope tag at the carboxylterminus. The MBP and 6X His tags permit purification of the recombinant protein from induced bacteria with the appropriate affinity matrix and allow recognition of the fusion protein with anti-MBP and anti-His antibodies. The 6X His epitope tag is generated by adding 6 histidine codons to the 3' cloning primer. A Factor Xa recognition site permits cleavage of the pMAL tag from 109P1D4. The pMAL-c2X and pMAL-p2X vectors are optimized to express the recombinant protein in the cytoplasm or periplasm respectively. Periplasm expression enhances folding of proteins with disulfide bonds. In one embodiment, amino acids 24-419 of 109P1D4 variant 1 was cloned into the pMAL-c2X vector and was used to express the fusion protein.

pET Constructs: To express 109P1D4 in bacterial cells, all or parts of the 109P1D4 cDNA protein coding sequence are cloned into the pET family of vectors (Novagen, Madison, WI). These vectors allow tightly controlled expression of recombinant 109P1D4 protein In bacteria with and without fusion to proteins that enhance solubility, such as NusA and thioredoxin (Trx), and epitope tags, such as 6X His and S-Tag T that aid purification and detection of the recombinant protein. For example, constructs are made utilizing pET NusA fusion system 43.1 such that regions of the 109P1D4 protein are expressed as amino-terminal fusions to NusA. In 2 embodiments, amino acids 24-419 and 24-815 were loned into pET43.1 vector and used to express the fusion protein.

C. Yeast Constructs: pESC Constructs: To express 109P1 D4 in the yeast species Saccharomyces cerevislae for generation of recombinant protein and functional studies, all or parts of the 109P1D4 cDNA protein coding sequence are cloned into the pESC family of vectors each of which contain I of 4 selectable markers, HIS3, TRP1, LEU2, and URA3 (Stratagene, La Jolla, CA). These vectors allow controlled expression from the same plasmid of up to 2 different genes or cloned sequences containing either FagTu or Myc epitope tags in the same yeast cell. This system is useful to confirm protein-protein interactions of 109P1D4. In addition, expression in yeast yields similar post-translational modifications, such as glycosylations and phosphorylations, that are found when expressed in eukaryotic cells.

pESP Constructs: To express 109P1 D4 in the yeast spedes Saccharomyces pombe, all or parts of the 109P1 D4 cDNA protein coding sequence are cloned into the pESP family of vectors. These vectors allow controlled high level of 89 00 expression of a 109P1D4 protein sequence that is fused at either the amino terminus or at the carboxyl terminus to GST

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which aids purification of the recombinant protein. A FlagTepitope tag allows detection of the recombinant protein with anti- C Flag antibody.

C Example 8: Production of Recombinant 109P1D4 In Higher Eukaryotic Systems SA. Mammalian Constructs: To express recombinant 109P1D4 in eukaryotic cells, the full or partial length 109P1D4 cDNA sequences were cloned into any one of a variety of expression vectors known in the art One or more of the following regions of 109P1D4 Swere expressed in these constructs, amino adds 1 to 1021 or any 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 0 24, 25, 26, 27, 28, 29, 30 or more contiguous amino acids from 109P1D4 v.1; amino acids 1 to 1054, 1 to 1347, 1 to 1337, 1 to 1310, 1 to 1037, 1 to 1048, 1 to 1340 of v.2, v.3, v.4, v.5, v.6, v.7, and v.8 respectively; or any 8, 9, 10, 11, 12, 13, 14, c 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more contiguous amino acids from 109P1D4 variants, or analogs 0 thereof.

0 The constructs can be transfected into any one of a wide variety of mammalian cells such as 293T cells.

Transfected 293T cell lysates can be probed with the anti-109P1D4 polyconal serum, described herein.

pcDNA4/HisMax Constructs: To express 109P1D4 in mammalian cells, a 109P1D4 ORF, or portions thereof, of 109P1D4 are cloned into pcDNA4/HisMax Version A (Invitrogen, Carlsbad, CA). Protein expression is driven from the cytomegalovirus (CMV) promoter and the SP16 translational enhancer. The recombinant protein has Xpress M and six histidine (6X His) epitopes fused to the amino-terminus. The pcDNA4/HisMax vector also contains the bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability along with the origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen. The Zeocin resistance gene allows for selection of mammalian cells expressing the protein and the ampicillin resistance gene and ColE1 origin permits selection and maintenance of the plasmid in E coli.

pcONA3.1/Mvclis Constructs: To express 109P1D4 in mammalian cells, a 109P1D4 ORF, or portions thereof, of 109P1D4 with a consensus Kozak translation initiation site was cloned into pcDNA3.1/MycHis Version A (Invitrogen, Carlsbad, CA). Protein expression is driven from the cytomegalovirus (CMV) promoter. The recombinant proteins have the myc epitope and 6X His epitope fused to the carboxyl-terminus. The pcDNA3.1/MycHis vector also contains the bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability, along with the SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen. The Neomycin resistance gene can be used, as it allows for selection of mammalian cells expressing the protein and the ampidllin resistance gene and ColE1 origin permits selection and maintenance of the plasmid in E. cofi.

The complete ORF of 109P1D4 v.1 was cloned into the pcDNA3.1/MycHis construct to generate 109P1D4.pcDNA3.1/MycHis.

pcDNA3.1/CT-GFP-TOPO Construct: To express 109P1D4 in mammalian cells and to allow detection of the recombinant proteins using fluorescence, a 109P1D4 ORF, or portions thereof, with a consensus Kozak translation initiation site are cloned into pcDNA3.1/CT-GFP-TOPO (Invitrogen, CA). Protein expression is driven from the cytomegalovirus (CMV) promoter. The recombinant proteins have the Green Fluorescent Protein (GFP) fused to the carboxyl-terminus facilitating noninvasive, In viv detection and cell biology studies. The pcDNA3.1CT-GFP-TOPO vector also contains the bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability along with the SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen. The 0 0 Neomycin resistance gene allows for selection of mammalian cells that express the protein, and the ampicillin resistance Sgene and ColE1 origin permits selection and maintenance of the plasmid in E. colli. Additional constructs with an aminoterminal GFP fusion are made in pcDNA3.1/NT-GFP-TOPO spanning the entire length of a 109P1D4 protein.

SPAPta: A 109P1D4 ORF, or portions thereof, is cloned into pAPtag-5 (GenHunter Corp. Nashville, TN). This construct generates an alkaline phosphatase fusion at the carboxyl-terminus of a 109P1D4 protein while fusing the IgGi Ssignal sequence to the amino-terminus. Constructs are also generated in which alkaline phosphatase with an aminoterminal IgGic signal sequence Is fused to the amino-terminus of a 109P1D4 protein. The resulting recombinant 109P1D4 Sproteins are optimized for secretion into the media of transfected mammalian cells and can be used to identify proteins such C as ligands or receptors that interact with 109P1D4 proteins. Protein expression is driven from the CMV promoter and the O~ recombinant proteins also contain myc and 6X His epitopes fused at the carboxyl-terminus that facilitates detection and purification. The Zeocin resistance gene present in the vector allows for selection of mammalian cells expressing the 00 recombinant protein and the ampicillin resistance gene permits selection of the plasmid in E coli.

A 109P1D4 ORF, or portions thereof, were cloned into pTag-5. This vector is similar to pAPtag but ri without the alkaline phosphatase fusion. This construct generated 109P1D4 protein with an amino-terminal IgGK signal sequence and myc and 6X His epitope tags at the carboxyl-terminus that facilitate detection and affinity purification. The resulting recombinant 109P1D4 protein was optimized for secretion Into the media of transfected mammalian cells, and was used as immunogen or ligand to identify proteins such as ligands or receptors that interact with the 109P1D4 proteins.

Protein expression is driven from the CMV promoter. The Zeocin resistance gene present in the vector allows for selection of mammalian cells expressing the protein, and the ampicillin resistance gene permits selection of the plasmid in E. coli.

PsecFc: A 109P1D4 ORF, or portions thereof, is also cloned into psecFc. The psecFc vector was assembled by cloning the human immunoglobulin G1 (IgG) Fc (hinge, CH2, CH3 regions) into pSecTag2 (Invitrogen, California). This construct generates an IgG1 Fc fusion at the carboxyl-terminus of the 109P1 D4 proteins, while fusing the IgGK signal sequence to N-terminus. 109P1D4 fusions utilizing the murine IgG1 Fc region are also used. The resulting recombinant 109P104 proteins are optimized for secretion into the media of transfected mammalian cells, and can be used as immunogens or to identify proteins such as figands or receptors that interact with 109P1 D4 protein. Protein expression is driven from the CMV promoter. The hygromycin resistance gene present in the vector allows for selection of mammalian cells that express the recombinant protein, and the ampicilln resistance gene permits selection of the plasmid in E. coi.

pSRa( Constructs: To generate mammalian cell lines that express 109P1 D4 constitutively, 109P1D4 ORF, or portions thereof, were cloned into pSRa constructs. Amphotropic and ecotropic retroviruses were generated by transfection of pSRa constructs into the 293T-10A1 packaging line or co-transfection of pSRa and a helper plasmid (containing deleted packaging sequences) into the 293 cells, respectively. The retrovirus is used to infect a variety of mammalian cell Fnes, resulting in the Integration of the cloned gene, 109P1D4, into the host cell-lines. Protein expression Is driven from a long terminal repeat (LTR). The Neomycin resistance gene present in the vector allows for selection of mammalian cells that express the protein, and the ampidllin resistance gene and ColE1 origin permit selection and maintenance of the plasmid in E coli. The retroviral vectors can thereafter be used for infection and generation of various cell lines using, for example, PC3, NIH 3T3, TsuPrl, 293 or rat-1 cells.

Additional pSRa constructs are made that fuse an epitope tag such as the FLAGT tag to the carboxyl-terminus of 109PiD4 sequences to allow detection using anti-Rag antibodies. For example, the FLAG T sequence 5' GAT TAC AAG GAT GAC GAC GAT AAG 3' (SEQ ID NO: 56) is added to cloning primer at the 3' end of the ORF. Additional pSRa constructs are made to produce both amino-terminal and carboxyl-terminal GFP and myc/6X His fusion proteins of the fulllength 109P1D4 proteins.

1 00 Additional Viral Vectors: Additional constructs are made for viral-mediated delivery and expression of 109P1 D4.

SHigh virus titer leading to high level expression of 109P1D4 is achieved in viral delivery systems such as adenoviral vectors and herpes amplicon vectors. A 109P1D4 coding sequence or fragments thereof are amplified by PCR and subcloned Into the AdEasy shuttle vector (Stratagene). Recombination and virus packaging are performed according to the manufacturers J instructions to generate adenoviral vectors. Alternatively, 109P1D4 coding sequences or fragments thereof are cloned into v the HSV-1 vector (Imgenex) to generate herpes viral vectors. The viral vectors are thereafter used for infection of various cell lines such as PC3, NIH 3T3, 293 or rat-1 cells.

Regulated Expression Systems: To control expression of 109P1 D4 in mammalian cells, coding sequences of 109P1D4, or portions thereof, are cloned into regulated mammalian expression systems such as the T-Rex System S(Invitrogen), the GeneSwitch System (Invitrogen) and the tightly-regulated Ecdysone System (Stratagene). These systems allow the study of the temporal and concentration dependent effects of recombinant 109P1 D4. These vectors are thereafter 00 used to control expression of 109P1D4 in various cell lines such as PC3, NIH 3T3, 293 or rat-1 cells.

SB. Baculovlrus Expression Systems C- To generate recombinant 109P1D4 proteins in a baculovirus expression system, 109P1D4 ORF, or portions thereof, are cloned Into the baculovirus transfer vector pBlueBac 4.5 (Invitrogen), which provides a His-tag at the N-terminus.

Specifically, pBlueBac-109P1D4 is co-transfected with helper plasmid pBac-N-Blue (Invitrogen) into SF9 (Spodoptera frugiperda) insect cells to generate recombinant baculovirus (see Invilrogen Instruction manual for details). Baculovirus is then collected from cell supernatant and purified by plaque assay.

Recombinant 109P1D4 protein is then generated by infection of HighFive insect cells (Invitrogen) with purified baculovirus. Recombinant 109P104 protein can be detected using anti-109P1D4 or anti-His-tag antibody. 109P104 protein can be purified and used in various cell-based assays or as immunogen to generate polyconal and monoclonal antibodies specific for 109P1D4.

Example 9: Antaenlcity Profiles and Secondary Structure Figure(s) 5A-I, Figure 6A-I, Figure 7A-1, Figure 8A-I, and Figure 9A-I depict graphically five amino acid profiles of 109P1D4 variants 1 through 9, each assessment available by accessing the ProtScale website located on the World Wide Web at (.expasy.chlcgi-bln/protscale.pl) on the ExPasy molecular biology server.

These profiles: Figure 5, Hydrophllicity, (Hopp Woods K.R, 1981. Proc. Nail. Acad. Sd. U.S.A. 78:3824- 3828); Figure 6, Hydropathlcity, (Kyte Doolittle 1982. J. Mol. Biol. 157:105-132); Figure 7, Percentage Accessible Residues (Janin 1979 Nature 277:491-492); Figure 8, Average Flexibility, (Bhaskaran R, and Ponnuswamy 1988.

Int J. Pept Protein Res. 32:242-255); Figure 9, Beta-turn (Deleage, Roux B. 1987 Protein Engineering 1:289-294); and optionally others available in the art, such as on the ProtScale website, were used to identify antigenic regions of each of the 109P1D4 variant proteins. Each of the above amino acid profiles of 109P1D4 variants were generated using the following ProtScale parameters for analysis: 1) A window size of 9; 2) 100% weight of the window edges compared to the window center; and, 3) amino acid profile values normalized to lie between 0 and 1.

Hydrophilicity (Figure Hydropathicity (Figure 6) and Percentage Accessible Residues (Figure 7) profiles were used to determine stretches of hydrophilic amino acids values greater than 0.5 on the Hydrophilicity and Percentage Accessible Residues profile, and values less than 0.5 on the Hydropathicity profile). Such regions are likely to be exposed to the aqueous environment, be present on the surface of the protein, and thus available for immune recognition, such as by antibodies.

Average Flexibility (Figure 8) and Beta-turn (Fgure 9) profiles determine stretches of amino adds values 00 greater than 0.5 on the Beta-turn profile and the Average Flexibility profile) that are not constrained in secondary structures O such as beta sheets and alpha helices. Such regions are also more likely to be exposed on the protein and thus accessible to immune recognition, such as by antibodies.

Antigenic sequences of the 109P1D4 variant proteins indicated, by the profiles set forth in Figure 5, Figure 6, SFigure 7, Figure 8, and/or Figure 9 are used to prepare immunogens, either peptides or nucleic acids that encode them, to Sgenerate therapeutic and diagnostic anti-109P1D4 antibodies. The immunogen can be any 5, 6, 7, 8, 9, 10,11,12,13,14, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35,40,45, 50 or more than 50 contiguous amino adds, or the corresponding nucleic adds that encode them, from the 109P1D4 protein variants listed in Figures 2 and 3. In particular, peptide Simmunogens of the invention can comprise, a peptide region of at least 5 amino acids of Figures 2 and 3 in any whole 0 number increment that includes an amino add position having a value greater than 0.5 in the Hydrophilicity profiles of Figure a peptide region of at least 5 amino acids of Figures 2 and 3 in any whole number increment that indudes an amino acid 00 position having a value less than 0.5 in the Hydropathicity profile of Figure 6; a peptide region of at least 5 amino adds of SFigures 2 and 3 in any whole number increment that includes an amino add position having a value greater than 0.5 in the Percent Accessible Residues profiles of Figure 7; a peptide region of at least 5 amino adds of Figures 2 and 3 in any whole number increment that includes an amino add position having a value greater than 0.5 in the Average Flexibility profiles on Figure 8; and, a peptide region of at least 5 amino adds of Figures 2 and 3 in any whole number increment that includes an amino acid position having a value greater than 0.5 in the Beta-turn profile of Figures 9. Peptide immunogens of the invention can also comprise nucleic adds that encode any of the forgoing.

All immunogens of the invention, peptide or nucleic acid, can be embodied in human unit dose form, or comprised by a composition that includes a pharmaceutical exdpient compatible with human physiology.

The secondary structure of 109P1D4 protein variants, namely the predicted presence and location of alpha helices, extended strands, and random coils, are predicted from the primary amino acid sequence using the HNN Hierarchical Neural Network method (NPS@: Network Protein Sequence Analysis TIBS 2000 March Vol. 25, No 3 [2911:147-150 Combet Blanchet Geourjon C. and DelBage httpJ/pbil.ibcp.fr/cgi-binnpsa_automat.pl?page=npsann.html), accessed from the ExPasy molecular biology server located on the World Wide Web at (www.expasy.ch/tools/). This analysis for protein variants 1 through 9 are shown in Figure 13A through 131 respectively. The percent of structure for each variant comprised of alpha helix, extended strand, and random coil is also indicated.

Analysis for the potential presence of transmembrane domains in 109P1 D4 variant proteins was carried out using a variety of transmembrane prediction algorithms accessed from the ExPasy molecular biology server located on the World Wide Web at (www.expasy.ch/tools/). Shown graphically in figures 13J-R are the results of analyses using the TMpred program (top panels) and the TMHMM program (bottom panels) of 109P1D4 protein variants 1 through 9 respectively.

Analyses of the variants using other structural prediction programs are summarized in Table VI and Table L Example 10: Generation of 109P1D4 Polvclonal Antbodies Polyclonal antibodies can be raised in a mammal, for example, by one or more injections of an immunizing agent and, If desired, an adjuvant Typically, the immunizing agent andlor adjuvant will be injected in the mammal by multiple subcutaneous or intraperitoneal injections. In addition to immunizing with a full length 109P1D4 protein variant, computer algorithms are employed In design of Immunogens that based on amino add sequence analysis contain characteristics of being antigenic and available for recognition by the immune system of the immunized host (see the Example entitled "Antigenidty Profiles and Secondary Structure). Such regions would be predicted to be hydrophilic, flexible, in beta-turn conformations, and be exposed on the surface of the protein (see, Figure 5, Figure 6, Figure 7, Figure 8, or Figure 9 for 00 amino acid profiles that indicate such regions of 109P1D4 protein variant 1).

For example, recombinant bacterial fusion proteins or peptides containing hydrophilic, flexible, beta-tur regions of S109P1D4 protein variants are used as antigens to generate polyclonal antibodies in New Zealand White rabbits or monoclonal antibodies as described In the example entitled "Generation of 109PiD4 Monoclonal Antibodies (mAbs)'. For C/1 example, in 109P1D4 variant 1, such regions indude, but are not limited to, amino acids 22-39, amino acids 67-108, amino Sadds 200-232, amino acids 454-499, amino acids 525-537, amino adds 640-660, amino adds 834-880, and amino adds 929-942. It is useful to conjugate the immunizing agent to a protein known to be immunogenic in the mammal being immunized. Examples of such immunogenic proteins include, but are not limited to, keyhole limpet hemocyanin (KLH), serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. In 2 embodiments, peptides encoding amino adds 77-90 0 and amino adds 929-942 of 109P1D4 variant 1 were synthesized, conjugated to KLH, and used to immunize separate rabbits. Alternatively the immunizing agent may include all or portions of the 109P1D4 variant proteins, analogs or fusion proteins thereof. For example, the 109P1D4 variant 1 amino acid sequence can be fused using recombinant DNA 00 Stechniques to any one of a variety of fusion protein partners that are well known in the art, such as glutathione-S-transferase (GST) and HIS tagged fusion proteins. In 1 embodiment, amino adds 24-419 of 109P1D4 variant 1 was fused to NUSa using recombinant techniques and the pET43.1 expression vector, expressed, purified and used to immunize a rabbit Such fusion proteins are purified from induced bacteria using the appropriate affinity matrix.

Other recombinant bacterial fusion proteins that may be employed include maltose binding protein, LacZ, thioredoxin, NusA, or an immunoglobulin constant region (see the section entitled 'Production of 109P1D4 in Prokaryotic Systems" and Current Protocols In Molecular Biology, Volume 2, Unit 16, Frederick M. Ausubul et al. eds., 1995; Lnsley, Brady, Umes, Grosmaire, L, Damle, and Ledbetter, J.(1991) J.Exp. Med. 174, 561-566).

In addition to bacterial derived fusion proteins, mammalian expressed protein antigens are also used. These antigens are expressed from mammalian expression vectors such as the Tag5 and Fc-fusion vectors (see the section entitled "Production of Recombinant 109P1D4 in Eukaryotic Systems'), and retain post-translational modifications such as glycosylations found in native protein. In one embodiment, amino adds 24-812 of 109PID4 variant 1 was cloned Into the mammalian secretion vector, and expressed in 293T cells (See Figure 20). The recombinant protein is purified by metal chelate chromatography from tissue culture supematants of 293T cells stably expressing the recombinant vector. The purified Tag5 109P1D4 protein is then used as immunogen.

During the immunization protocol, it is useful to mix or emulsify the antigen in adjuvants that enhance the immune response of the host animal. Examples of adjuvants include, but are not limited to, complete Freund's adjuvant (CFA) and MPL-TDM adjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate).

In a typical protocol, rabbits are initially immunized subcutaneously with up to 200 pg, typically 100-200 pg, of fusion protein or peptide conjugated to KLH mixed in complete Freund's adjuvant (CFA). Rabbits are then injected subcutaneously every two weeks with up to 200 pg, typically 100-200 pg, of the immunogen in incomplete Freund's adjuvant (IFA). Test bleeds are taken approximately 7-10 days following each immunization and used to monitor the titer of the antiserum by ELISA.

To test reactivity and specificity of immune serum, such as the rabbit serum derived from immunization with the NUSa-fusion of 109P1D4 variant 1 protein, the full-length 109P1D4 variant 1 cDNA is cloned into pCDNA 3.1 myo-his expression vector (Invitrogen, see the Example entitled "Production of Recombinant 109P1D4 in Eukaryotic Systems'). After Iransfection of the constructs into 293T cells, cell lysates are probed with the anti-109PD4 serum to determine specific reactivity to denatured 109P1D4 protein using the Western blot technique. Probing with anti-His antibody serves as a positive control for expression of 109P1D4 in the transfected cells (See Figure 21). In addition, the immune serum is tested 00 by fluorescence microscopy, flow cytometry and immunoprecipitation against 293T and other recombinant 109P1D4- Sexpressing cells to determine specific recognition of native protein. Western blot immunoprecipitation, fluorescent microscopy, and flow cytometric techniques using cells that endogenously express 109P1 D4 are also carried out to test 1) reactivity and specificity.

Anti-serum from rabbits immunized with 109P104 variant fusion proteins, such as GST and MBP fusion proteins, are purified by depletion of antibodies reactive to the fusion partner sequence by passage over an affinity column containing the fusion partner either alone or in the context of an irrelevant fusion protein. For example, antiserum derived from a NUSa- 109P1D4 variant 1 fusion protein is first purified by passage over a column of MBP protein covalently coupled to AffiGel C matrix (BioRad, Hercules, Calif.). The antiserum is then affinity purified by passage over a column composed of a NUSa- S109P1D4 fusion protein covalently coupled to Affigel matrix. The serum is then further purified by protein G affinity chromatography to isolate the IgG fraction. Sera from other His-tagged antigens and peptide immunized rabbits as well as 00 fusion partner depleted sera are affinity purified by passage over a column matrix composed of the original protein Simmunogen or free peptide.

Example 11: Generation of 109PD4 Monodonal Antibodies (mAbs) In one embodiment, therapeutic mAbs to 109P1 D4 variants comprise those that react with epitopes specific for each variant protein or specific to sequences in common between the variants that would disrupt or modulate the biological function of the 109P1D4 variants, for example those that would disrupt the interaction with ligands and binding partners.

Immunogens for generation of such mAbs include those designed to encode or contain the entire 109P1D4 protein variant sequence, regions predicted to contain functional motifs, and regions of the 109P1D4 protein variants predicted to be antigenic from computer analysis of the amino add sequence (see, Figure 5, Figure 6, Figure 7, Figure 8, or Figure 9, and the Example entitled 'Antigenidty Profiles and Secondary Structure). Immunogens include peptides, recombinant bacterial proteins, and mammalian expressed Tag 5 proteins and human and murine IgG FC fusion proteins. In addition, cells engineered to express high levels of a respective 109P1D4 variant, such as 293T-109P1D4 variant 1 or 300.19- 109P1D4 variant 1murine Pre-B cells, are used to immunize mice.

To generate mAbs to a 109P104 variant, mice are first immunized Intraperitoneally (IP) with, typically, 10-50 pg of protein immunogen or 107 109P1D4-expressing cells mixed in complete Freund's adjuvant Mice are then subsequently immunized IP every 2-4 weeks with, typically, 10-50 pg of protein immunogen or 107 cells mixed in incomplete Freund's adjuvant Alternatively, MPL-TDM adjuvant is used in immunizations. In addition to the above protein and cell-based immunization strategies, a DNA-based immunization protocol is employed in which a mammalian expression vector encoding a 109P1D4 variant sequence is used to immunize mice by direct injection of the plasmid DNA. For example, amino adds 24-812 of 109P1D4 of variant 1 Is cloned into the Tag5 mammalian secretion vector and the recombinant vector will then be used as immunogen. In another example the same amino acids are cloned into an Fc-fusion secretion vector in which the 109P1D4 variant 1 sequence is fused at the amino-terminus to an IgK leader sequence and at the carboxylterminus to the coding sequence of the human or murine IgG Fc region. This recombinant vector is then used as immunogen. The plasmid immunization protocols are used in combination with purified proteins expressed from the same vector and with cells expressing the respective 109P1D4 variant Alternatively, mice may be immunized directly into their footpads. In this case, 10-50 pg of protein immunogen or 107 254P1D6B-expressing cells are injected sub-cutaneously into the footpad of each hind leg. The first immunization is given with Titermax (Sigma T M as an adjuvant and subsequent injections are given with Alum-gel in conjunction with CpG oligonudeotide sequences with the exception of the final injection which is given with PBS. Injections are given twice weekly 00 0 (every three to four days) for a period of 4 weeks and mice are sacrificed 3-4 days after the final injection, at which point Slymph nodes immediately draining from the footpad are harvested and the B-cells are collected for use as antibody producing fusion partners.

During the immunization protocol, test bleeds are taken 7-10 days following an injection to monitor titer and specificity of the immune response. Once appropriate reactivity and specificity is obtained as determined by ELISA, Western 0 blotting, immunoprecpitation, fluorescence microscopy, and flow cytometric analyses, fusion and hybridoma generation is then carried out with established procedures well known in the art (see, Harlow and Lane, 1988).

In one embodiment for generating 109P1 D4 monoclonal antibodies, a Tag5 antigen of variant 1 encoding amino C1 acids 14-812 is expressed in 293T cells and purified from conditioned media. Balb C mice are initially immunized C- intraperitoneally with 25 gg of the Tag5 109P1 D4 variant 1 protein mixed in complete Freund's adjuvant Mice are Ssubsequently Immunized every two weeks with 25 pg of the antigen mixed In Incomplete Freund's adjuvant for a total of 00 three immunizations. ELISA using the Tag5 antigen determines the titer of serum from immunized mice. Reactivity and specificity of serum to full length 109P1 D4 variant 1 protein is monitored by Westem blotting, Immunoprecpitation and flow

C

cytometry using 293T cells transfected with an expression vector encoding the 109P1D4 variant 1 cDNA (see the Example entitled 'Production of Recombinant 109P1D4 in Higher Eukaryotic Systems" and Figure 21). Other recombinant 109P1D4 variant 1-expressing cells or cells endogenously expressing 109P1D4 variant 1 are also used. Mice showing the strongest reactivity are rested and given a final Injection of antigen in PBS and then sacrificed four days later. The spleens of the sacrificed mice are harvested and fused to SPO/2 myeloma cells using standard procedures (Harlow and Lane, 1988).

Supematants from HAT selected growth wells are screened by EUSA, Western blot, immunoprecipitation, fluorescent microscopy, and flow cytometry to Identify 109P1D4 specific antibody-producing clones.

To generate monoconal antibodies that are specific for a 109P1D4 variant protein, immunogens are designed to encode sequences unique for each variant In one embodiment, an antigenic peptide composed of amino acids 1-29 of 109P1D4 variant 2 is coupled to KLH to derive monoclonal antibodies specific to 109P1D4 variant 2. In another embodiment, an antigenic peptide comprised of amino acids 1-23 of 109P1D4 variant 6 is coupled to KLH and used as immunogen to derive varaiant 6 specific MAbs. In another example, a GST-fuslon protein encoding amino adds 1001-1347 of variant 3 is used as immunogen to generate antibodies that would recognize variants 3, 4, 5, and 8, and distinguish them from variants 1, 2, 6, 7and 9. Hybridoma supematants are then screened on the respective antigen and then further screened on cells expressing the specific variant and cross-screened on cells expressing the other variants to derive variantspecific monodonal antibodies.

The binding affinity of 109P1D4 variant specific monodonal antibodies are determined using standard technologies. Affinity measurements quantify the strength of antibody to epitope binding and are used to help define which 109P1D4 variant monodonal antibodies preferred for diagnostic or therapeutic use, as appreciated by one of skill in the art The BIAcore system (Uppsala, Sweden) is a preferred method for determining binding affinity. The BlAcore system uses surface plasmon resonance (SPR, Welford K. 1991, Opt Quant Elect 23:1; Morton and Myszka, 1998, Methods in Enzymology 295: 268) to monitor blomolecular interactions in real time. BlAcore analysis conveniently generates association rate constants, dissociation rate constants, equilibrium dissociation constants, and affinity constants.

Alternatively, equilibrium binding analysis of MAbs on 109P1D4-expressing cells can be used to determine affinity.

Example 12: HLA Class I and Class II Binding Assays HLA class I and class II binding assays using purified HLA molecules are performed in accordance with disdosed protocols PCT publications WO 94/20127 and WO 94/03205; Sidney et al., Current Profocols In Immunology 18.3.1 00 O (1998); Sidney, et aL, J. Immunol. 154:247 (1995); Sette, et al., Mol. Immunol. 31:813 (1994)). Briefly, purified MHC molecules (5 to 500 nM) are incubated with various unlabeled peptide inhibitors and 1-10 nM 1 5 -radiolabeled probe o peptides as described. Following incubation, MHC-peptide complexes are separated from free peptide by gel filtration and the fraction of peptide bound is determined. Typically, in preliminary experiments, each MHC preparation Is titered in the presence of fixed amounts of radiolabeled peptides to determine the concentration of HLA molecules necessary to bind of the total radioactivity. All subsequent inhibition and direct binding assays are performed using these HLA concentrations.

Since under these conditions [abel]<[HLA] and ICso.[HLA], the measured ICso values are reasonable approximations of the true KD values. Peptide Inhibitors are typically tested at concentrations ranging from 120 pg/ml to 1.2 r ng/ml, and are tested in two to four completely independent experiments. To allow comparison of the data obtained in C= different experiments, a relative binding figure is calculated for each peptide by dividing the ICso of a positive control for 00 Inhibition by the ICso for each tested peptide (typically unlabeled versions of the radiolabeled probe peptide). For database Spurposes, and inter-experiment comparisons, relative binding values are compiled. These values can subsequently be converted back Into ICso nM values by dividing the ICso nM of the positive controls for inhibition by the relative binding of the peptide of interest This method of data compilation Is accurate and consistent for comparing peptides that have been tested on different days, or with different lots of purified MHC.

Binding assays as outlined above may be used to analyze HLA supermotif and/or HLA motif-bearing peptides (see Table IV).

Example 13: Identification of HLA Supermotif- and Motif-Bearing CTL Candidate Epitopes HLA vaccine compositions of the invention can include multiple epitopes. The multiple epitopes can comprise multiple HLA supermotifs or motifs to achieve broad population coverage. This example illustrates the identification and confirmation of supermotif- and motif-bearing epitopes for the inclusion in such a vaccine composition. Calculation of population coverage is performed using the strategy described below.

Computer searches and algorithms for identification of supermotif and/or molif-bearing epitopes The searches performed to identify the motif-bearing peptide sequences in the Example entitled "Antigenilty Profiles" and Tables VIII-XXI and XXII-XLIX employ the protein sequence data from the gene product of 109P1D4 set forth In Figures 2 and 3, the specific search peptides used to generate the tables are listed in Table VII.

Computer searches for epitopes bearing HLA Class I or Class II supermotifs or motifs are performed as follows.

All translated 109P1D4 protein sequences are analyzed using a text string search software program to identify potential peptide sequences containing appropriate HLA binding motifs; such programs are readily produced in accordance with information in the art in view of known motif/supermotif disclosures. Furthermore, such calculations can be made mentally.

Identified A2-, A3-, and DR-supermotif sequences are scored using polynomial algorithms to predict their capacity to bind to specific HLA-Class I or Class II molecules. These polynomial algorithms account for the impact of different amino adds at different positions, and are essentially based on the premise that the overall affinity (or AG) of peptide-HLA molecule interactions can be approximated as a linear polynomlal function of the type: "AG' ax a2X xa where aV is a coefficient which represents the effect of the presence of a given amino add at a given position along the sequence of a peptide of n amino acids. The crucial assumption of this method is that the effects at each position are essentially independent of each other independent binding of Individual side-chains). When residue j occurs at position I in the peptide, it is assumed to contribute a constant amount ]i to the free energy of binding of the peptide 97 00 irrespective of the sequence of the rest of the peptide.

The method of derivation of specific algorithm coefficients has been described in Gulukota et al., J. Mol. Biol.

C1 267:1258-126,1997; (see also Sidney et al., Human Immunol. 45:79-93,1996; and Southwood et J. Immunol. 160:3363- 3373, 1998). Briefly, for all I positions, anchor and non-anchor alike, the geometric mean of the average relative binding y) (ARB) of all peptides carrying jis calculated relative to the remainder of the group, and used as the estimate ofj For Class SII peptides, if multiple alignments are possible, only the highest scoring alignment Is utilized, following an iterative procedure.

To calculate an algorithm score of a given peptide in a test set, the ARB values corresponding to the sequence of the peptide are multiplied. If this product exceeds a chosen threshold, the peptide is predicted to bind. Appropriate thresholds are Schosen as a function of the degree of stringency of prediction desired.

Selection of HLA-A2 supertvpe cross-reactive peptides C Protein sequences from 109P1D4 are scanned utilizing motif Identification software, to identify 9- 10- and 11- 00 00 mer sequences containing the HLA-A2-supermotif main anchor specificity. Typically, these sequences are then scored using Sthe protocol described above and the peptides corresponding to the positive-scoring sequences are synthesized and tested for their capacity to bind purified HLA-A'0201 molecules in vtro (HLA-A*0201 is considered a prototype A2 supertype molecule).

These peptides are then tested for the capacity to bind to additional A2-supertype molecules (A*0202, A*0203, A*0206, and A*6802). Peptides that bind to at least three of the five A2-supertype alleles tested are typically deemed A2supertype cross-reactive binders. Preferred peptides bind at an affinity equal to or less than 500 nM to three or more HLA- A2 supertype molecules.

Selection of HLA-A3 supenrotif-bearing epitopes The 109P1D4 protein sequence(s) scanned above is also examined for the presence of peptides with the HLA-A3supermotif primary anchors. Peptides corresponding to the HLA A3 supermotif-bearing sequences are then synthesized and tested for binding to HLA-A*0301 and HLA-A*1101 molecules, the molecules encoded by the two most prevalent A3supertype alleles. The peptides that bind at least one of the two alleles with binding affinities of <500 nM, often 200 nM, are then tested for binding cross-reactivity to the other common A3-supertype alleles A'3101, A'3301, and A*6801) to identify those that can bind at least three of the five HLA-A3-supertype molecules tested.

Selection of HLA-B7 supermotif bearing epitoes The 109P1D4 protein(s) scanned above is also analyzed for the presence of 9- 10-, or 11-mer peptides with the HLA-87-supermotif. Corresponding peptides are synthesized and tested for binding to HLA-B80702, the molecule encoded by the most common B7-supertype allele the prototype B7 supertype allele). Peptides binding B'0702 with ICso of :500 nM are Identified using standard methods. These peptides are then tested for binding to other common B7-supertype molecules B'3501, B'5101, B*5301, and 8*5401). Peptides capable of binding to three or more of the five B7supertype alleles tested are thereby identified.

Selection of Al and A24 motif-bearing epitopes To further increase population coverage, HLA-A1 and -A24 epltopes can also be incorporated into vaccine compositions. An analysis of the 109P1D4 protein can also be performed to identify HLA-Ai- and A24-motif-containing sequences.

I 0 High affinity and/or cross-reactive binding epitopes that bear other motif and/or supermotifs are identified using C analogous methodology.

S Example 14: Confirmation of Immunogenicity Cross-reactive candidate CTL A2-supermotif-bearing peptides that are identified as described herein are selected 0to confirm In vitro Immunogenicity. Confirmation is performed using the following methodology: Target Cell Unes for Cellular Screening: The .221A2.1 cell line, produced by transferring the HLA-A2.1 gene into the HLA-A, -C null mutant human B- Slymphoblastoid cell line 721.221, is used as the peptide-loaded target to measure activity of HLA-A2.1-restricted CTL This C cell line is grown in RPMI-1640 medium supplemented with antibiotics, sodium pyruvate, nonessential amino acids and heat inactivated FCS. Cells that express an antigen of interest, or transfectants comprising the gene encoding the 00 antigen of Interest, can be used as target cells to confirm the ability of peptide-specific CTLs to recognize endogenous antigen.

Primary CTL Induction Cultures: Generation of Dendritic Cells PBMCs are thawed in RPMI with 30 pg/ml DNAse, washed twice and resuspended in complete medium (RPMI-1640 plus 5% AB human serum, non-essential amino adds, sodium pyruvate, Lglutamlne and penicillinstreptomycin). The monocytes are purified by plating 10 x 108 PBMC/well In a 6-well plate. After 2 hours at 37°C, the non-adherent cells are removed by gently shaking the plates and aspirating the supematants. The wells are washed a total of three times with 3 ml RPMI to remove most of the non-adherent and loosely adherent cells. Three ml of complete medium containing 50 ng/ml of GM-CSF and 1,000 Ulml of IL-4 are then added to each well. TNFc is added to the DCs on day 6 at 75 ng/mn and the cells are used for CTL induction cultures on day 7.

Induction of CTL with DC and Peptide: CD8+ T-cells are isolated by positive selection with Dynal immunomagnetic beads (Dynabeads@ M-450) and the detacha-bead@ reagent Typically about 200-250x106 PBMC are processed to obtain 24x10s CD8 T-cels (enough for a 48-well plate culture). Briefly, the PBMCs are thawed in RPMI with 30pg/ml DNAse, washed once with PBS containing 1% human AB serum and resuspended In PBS/1% AB serum at a concentration of 20x10cells/ml. The magnetic beads are washed 3 times with PBS/AB serum, added to the cells (1 4 0 pl beads/20x10 6 cells) and incubated for 1 hour at 4 0 C with continuous mixing. The beads and cells are washed 4x with PBS/AB serum to remove the nonadherent cells and resuspended at 100x10 6 cells/ml (based on the original cell number) in PBS/AB serum containing 100pl/ml detacha-bead@ reagent and 30 pg/ml DNAse. The mixture is incubated for 1 hour at room temperature with continuous mixing. The beads are washed again with PBS/AB/DNAse to collect the CD8+ T-cells. The DC are collected and centrifuged at 1300 rpm for 5-7 minutes, washed once with PBS with 1% BSA, counted and pulsed with 40pg/ml of peptide at a cell concentration of 1-2x10s/ml in the presence of 3pg/ml B1- microglobulin for 4 hours at 20°C. The DC are then irradiated (4,200 rads), washed 1 time with medium and counted again.

Setting up Induction cultures: 0.25 ml cytokine-generated DC (at ix10 5 cells/ml) are co-cultured with 0.25ml of CD8+ T-cells (at 2x10 6 cell/ml) in each well of a 48-well plate in the presence of 10 ng/ml of IL-7. Recombinant human is added the next day at a final concentration of 10 ng/ml and rhuman IL-2 is added 48 hours later at 10 IU/ml.

Restimulation of the induction cultures with peptide-pulsed adherent cells: Seven and fourteen days after the primary induction, the cells are restimulated with peptide-pulsed adherent cells. The PBMCs are thawed and washed twice with RPMI and DNAse. The cells are resuspended at 5x106 cells/ml and irradiated at -4200 rads. The PBMCs are plated at 2x10 6 In 0.5 ml complete medium per well and incubated for 2 hours at 37°C. The plates are washed twice with RPMI by tapping the plate gently to remove the nonadherent cells and the adherent cells pulsed with 10pgiml of peptide in the 00 0 presence of 3 pg/ml 62 microglobulin in 0.25ml RPMI/5%AB per well for 2 hours at 37 0 C. Peptide solution from each well is Saspirated and the wells are washed once with RPMI. Most of the media is aspirated from the induction cultures (CD8+ cells) Sand brought to 0.5 ml with fresh media. The cells are then transferred to the wells containing the peptide-pulsed adherent cells. Twenty four hours later recombinant human IL-10 is added at a final concentration of 10 ng/ml and recombinant human 112 is added the next day and again 2-3 days later at 501Ulml (Tsai et al., Critical Reviews in Immunology S18(1-2):65-75, 1998). Seven days later, the cultures are assayed for CTL activity in a 5 'Cr release assay. In some experiments the cultures are assayed for peptide-specific recognition in the in situ IFNy ELISA at the time of the second restimulation followed by assay of endogenous recognition 7 days later. After expansion, activity is measured In both assays for a side-by-side comparison.

CK Measurement of CTL lytic activity by 5sCr release.

Seven days after the second restimulation, cytotoxicity is determined In a standard (5 hr) 5 'Cr release assay by 00 assaying individual wells at a single E:T. Peptide-pulsed targets are prepared by incubating the cells with 10pgIml peptide Sovernight at 37C.

Adherent target cells are removed from culture flasks with trypsin-EDTA Target cells are labeled with 200pCi of s Cr sodium chromate (Dupont, Wilmington, DE) for 1 hour at 37°C. Labeled target cells are resuspended at 106 per ml and diluted 1:10 with K562 cells at a concentration of 3.3x10/ml (an NK-sensitive erythroblastoma cell lne used to reduce nonspecific lysis). Target cells (100 pl) and effectors (100pl) are plated in 96 well round-bottom plates and incubated for 5 hours at 370C. At that time, 100 pl of supematant are collected from each well and percent lysis is determined according to the formula: [(cpm of the test sample- cpm of the spontaneous s 5 Cr release sample)/(cpm of the maximal 5 sCr release samplecpm of the spontaneous s5Cr release sample)] x 100.

Maximum and spontaneous release are determined by incubating the labeled targets with 1% Triton X-100 and media alone, respectively. A positive culture is defined as one in which the specific lysis (sample- background) is 10% or higher in the case of individual wells and is 15% or more at the two highest E:T ratios when expanded cultures are assayed.

In situ Measurement of Human IFNy Production as an Indicator of Peptide-specfic and Endogenous Recognition Immulon 2 plates are coated with mouse anti-human IFNy monoclonal antibody (4 pg/ml 0.1M NaHCO3, pH8.2) overnight at 4°C. The plates are washed with Ca 2 Mg 2 +-free PBS/0.05% Tween 20 and blocked with PBS/10% FCS for two hours, after which the CTLs (100 pi/well) and targets (100 pl/well) are added to each well, leaving empty wells for the standards and blanks (which received media only). The target cells, either peptide-pulsed or endogenous targets, are used at a concentration of lx106 cells/ml. The plates are incubated for 48 hours at 37°C with 5% CO2.

Recombinant human IFN-gamma is added to the standard wells starting at 400 pg or 1200pg/100 microliter/well and the plate incubated for two hours at 37°C. The plates are washed and 100 pJ of blotinylated mouse anti-human IFNgamma monoclonal antibody (2 microgram/ml In PBS/3%FCS/0.05% Tween 20) are added and incubated for 2 hours at room temperature. After washing again, 100 microliter HRP-streptavidin (1:4000) are added and the plates incubated for one hour at room temperature. The plates are then washed 6x with wash buffer, 100 microliter/well developing solution (TMB 1:1) are added, and the plates allowed to develop for 5-15 minutes. The reaction Is stopped with 50 microliter/well 1M H3P04 and read at OD450. A culture is considered positive if it measured at least 50 pg of IFN-gamma/well above background and is twice the background level of expression.

CTL Expansion.

Those cultures that demonstrate specific lytic activity against peptide-pulsed targets and/or tumor targets are expanded over a two week period with ani-CD3. Briefly, 5x1 0 CD8+ cells are added to a T25 flask containing the following: 00 S1x10 6 irradiated (4,200 rad) PBMC (autologous or allogeneic) per ml, 2x10 5 irradiated (8,000 rad) EBV- transformed cells per ml, and OKT3 (anti-CD3) at 30ng per ml in RPMI-1640 containing 10% human AB serum, non-essential amino adds, Ssodium pyruvate, 25pM 2-mercaptoethanol, L-glutamine and pencillin/streptomycin. Recombinant human IL2 is added 24 hours later at a final concentration of 2001U/ml and every three days thereafter with fresh media at 501U/ml. The cells are split if the cell concentration exceeds ix0S/ml and the cultures are assayed between days 13 and 15 at E:T ratios of 30, S3 and 1:1 in the s 5 Cr release assay or at Ixl0O/ml in the in situ IFNy assay using the same targets as before the expansion.

Cultures are expanded in the absence of anti-CD3* as follows. Those cultures that demonstrate specific lytic activity against peptide and endogenous targets are selected and 5x10 4 CD8+ cells are added to a T25 flask containing the following: lx106 autologous PBMC per ml which have been peptide-pulsed with 10 pg/ml peptide for two hours at 37*C and CKl irradiated (4,200 rad); 2x10 5 irradiated (8,000 rad) EBV-transformed cells per ml RPMI-1640 containing 10%(v/v) human AB C serum, non-essential AA, sodium pyruvate, 25mM 2-ME, L-glutamine and gentamicin.

00 Immunoaenicitv of A2 supernotif-bearing peptides SA2-supermotif cross-reactive binding peptides are tested in the cellular assay for the ability to induce peptidespecific CTL in normal individuals. In this analysis, a peptide Is typically considered to be an epitope if it induces peptidespecific CTLs in at least individuals, and preferably, also recognizes the endogenously expressed peptide.

Immunogenicity can also be confirmed using PBMCs isolated from patients bearing a tumor that expresses 109P1D4. Briefly, PBMCs are isolated from patients, re-stimulated with peptide-pulsed monocytes and assayed for the ability to recognize peptide-pulsed target cells as well as transfected cells endogenously expressing the antigen.

Evaluation of A*03/A11 Immunooenicity HLA-A3 supermotif-bearing cross-reactive binding peptides are also evaluated for Immunogenicity using methodology analogous for that used to evaluate the immunogenicity of the HLA-A2 supermotif peptides.

Evaluation of B7 immunoenicity Immunogenicity screening of the B7-supertype cross-reactive binding peptides identified as set forth herein are confirmed in a manner analogous to the confirmation of A2-and A3-supermotif-bearing peptides.

Peptides bearing other supermotifs/motifs, HLA-A1, HLA-A24 etc. are also confirmed using similar methodology Example 15: Implementation of the Extended Supennotf to Improve the Bindina Capacity of Native Epitopes by Creating Analos HLA motifs and supermotifs (comprising primary and/or secondary residues) are useful In the identification and preparation of highly cross-reactive native peptides, as demonstrated herein. Moreover, the definition of HLA motifs and supermotifs also allows one to engineer highly cross-reactive epitopes by identifying residues within a native peptide sequence which can be analoged to confer upon the peptide certain characteristics, e.g. greater cross-reactivity within the group of HLA molecules that comprise a supertype, and/or greater binding affinity for some or all of those HLA molecules.

Examples of analoging peptides to exhibit modulated binding affinity are set forth in this example.

Analooing at Primary Anchor Residues Peptide engineering strategies are implemented to further increase the cross-reactivity of the epitopes. For example, the main anchors of A2-supermotif-bearing peptides are altered, for example, to introduce a preferred L, I, V, or M at position 2, and I or V at the C-terminus.

To analyze the cross-reactivity of the analog peptides, each engineered analog is initially tested for binding to the prototype A2 supertype allele A*0201, then, ifA*0201 binding capacity is maintained, for A2-supertype cross-reactivity.

00 SAlternatively, a peptide is confirmed as binding one or all supertype members and then analoged to modulate O binding affinity to any one (or more) of the supertype members to add population coverage.

The selection of analogs for immunogenicity in a cellular screening analysis is typically further restricted by the 0) capacity of the parent wild type (WT) peptide to bind at least weakly, bind at an ICso of 5000nM or less, to three of more A2 supertype alleles. The rationale for this requirement is that the WT peptides must be present endogenously in sufficient Squantity to be biologically relevant Analoged peptides have been shown to have increased immunogenicity and crossreactivity by T cells specific for the parent epitope (see, Parkhurst et al., J. Immunol. 157:2539, 1996; and Pogue et al., Proc. Natl. Acad. Sci. USA 92:8166,1995).

C In the cellular screening of these peptide analogs, it is important to confirm that analog-specific CTLs are also able N to recognize the wild-type peptide and, when possible, target cells that endogenously express the epitope.

00 Analooing of HLA-A3 and B7-supermotif-bearing eptides Analogs of HLA-A3 supermotif-bearing epitopes are generated using strategies similar to those employed in CN analoging HLA-A2 supermotif-bearing peptides. For example, peptides binding to 3/5 of the A3-supertype molecules are engineered at primary anchor residues to possess a preferred residue S, M, or A) at position 2.

The analog peptides are then tested for the ability to bind A'03 and A*11 (prototype A3 supertype alleles). Those peptides that demonstrate 500 nM binding capacity are then confirmed as having A3-supertype cross-reactivity.

Similarly to the A2- and A3- motif bearing peptides, peptides binding 3 or more 87-supertype alleles can be improved, where possible, to achieve increased cross-reactive binding or greater binding affinity or binding half life. B7 supermotif-bearing peptides are, for example, engineered to possess a preferred residue I, L, or F) at the C-terminal primary anchor position, as demonstrated by Sidney et al. Immunol. 157:3480-3490, 1996).

Analoging at primary anchor residues of other motif and/or supermotif-bearing epitopes is performed in a like manner.

The analog peptides are then be confirmed for immunogenicity, typically in a cellular screening assay. Again, it is generally important to demonstrate that analog-specific CTLs are also able to recognize the wild-type peptide and, when possible, targets that endogenously express the epitope.

Analgilng at Secondary Anchor Residues Moreover, HLA supermotifs are of value in engineering highly cross-reactive peptides and/or peptides that bind HLA molecules with increased affinity by identifying particular residues at secondary anchor positions that are associated with such properties. For example, the binding capacity of a B7 supermotif-bearing peptide with an F residue at position 1 is analyzed. The peptde is then analoged to, for example, substitute L for F at position 1. The analoged peptide is evaluated for increased binding affinity, binding half life and/or increased cross-reactivity. Such a procedure identifies analoged peptides with enhanced properties.

Engineered analogs with sufficiently improved binding capacity or cross-reactivity can also be tested for Immunogenicity in HLA-B7-transgenlc mice, following for example, IFA Immunization or lipopeptide immunization. Analoged peptides are additionally tested for the ability to stimulate a recall response using PBMC from patients with 109P1D4expressing tumors.

Other analoging strategies Another form of peptide analoging, unrelated to anchor positions, Involves the substitution of a cysteine with oamino butyric acid. Due to its chemical nature, cysteine has the propensity to form disulfide bridges and sufficiently alter the 00 0 peptide structurally so as to reduce binding capacity. Substitution of a-amino butyric acid for cysteine not only alleviates this problem, but has been shown to improve binding and crossbinding capabilities in some instances (see, the review by SSette et al., In: Persistent Viral Infections, Eds. R Ahmed and I. Chen, John Wiley Sons, England, 1999).

Thus, by the use of single amino acd substitutions, the binding properties and/or cross-reactivity of peptide ligands for HLA supertype molecules can be modulated.

ExamPle 16: Identification and confirmation of 109P1D4-derived sequences with HLA-DR binding motifs Peptide epitopes bearing an HLA class II supermotif or motif are Identified and confirmed as outlined below using C methodology similar to that described for HLA Class I peptides.

lI .Selection of HLA-DR-supermotif-bearing eitopes.

STo identify 109P1D4-derived, HLA class II HTL epitopes, a 109P1 D4 antigen is analyzed for the presence of 00 sequences bearing an HLA-DR-motif or supermotif. Specifically, 15-mer sequences are selected comprising a DRsupermotif, comprising a 9-mer core, and three-residue N- and C-terminal flanking regions (15 amino adds total).

cN Protocols for predicting peptide binding to DR molecules have been developed (Southwood etal., J. Immunol.

160:3363-3373, 1998). These protocols, specific for individual DR molecules, allow the scoring, and ranking, of 9-mer core regions. Each protocol not only scores peptide sequences for the presence of DR-supermotif primary anchors at position 1 and position 6) within a 9-mer core, but additionally evaluates sequences for the presence of secondary anchors.

Using allele-specific selection tables (see, Southwood etal., Ibid.), it has been found that these protocols efficently select peptide sequences with a high probability of binding a particular DR molecule. Additionally, it has been found that performing these protocols in tandem, specifically those for DR1, DR4w4, and DR7, can efficiently select DR cross-reactive peptides.

The 109P1D4-derved peptides identified above are tested for their binding capacity for various common HLA-DR molecules. All peptides are initially tested for binding to the DR molecules in the primary panel: DR1, DR4w4, and DR7.

Peptides binding at least two of these three DR molecules are then tested for binding to DR2w2 p1, DR2w2 p2, DR6wl9, and DR9 molecules in secondary assays. Finally, peptides binding at least two of the four secondary panel DR molecules, and thus cumulatively at least four of seven different DR molecules, are screened for binding to DR4w15, DR5w11, and DR8w2 molecules in tertiary assays. Peptides binding at least seven of the ten DR molecules comprising the primary, secondary,and tertiary screening assays are considered cross-reactive DR binders. 109P1D4-derived peptides found to bind common HLA-DR alleles are of particular interest Selection of DR3 motif peptides Because HLA-DR3 is an allele that Is prevalent In Caucasian, Black, and Hispanic populations, DR3 binding capacity is a relevant criterion in the selection of HTL epitopes. Thus, peptides shown to be candidates may also be assayed for their DR3 binding capadty. However, in view of the binding specificty of the DR3 motif, peptides binding only to DR3 can also be considered as candidates for inclusion in a vaccine formulation.

To efficiently identify peptides that bind DR3, target 109P1D4 antigens are analyzed for sequences carrying one of the two DR3-speciflc binding motifs reported by Geluk et al. Immunol. 152:5742-5748, 1994). The corresponding peptides are then synthesized and confirmed as having the ability to bind DR3 with an affinity of 1pM or better, less than 1 pM. Peptides are found that meet this binding criterion and qualify as HLA class II high affinity binders.

DR3 binding epitopes identified in this manner are Included in vaccine compositions with DR supermotif-bearing peptide epitopes.

Similarly to the case of HLA class I motif-bearing peptides, the dass II motif-bearing peptides are analoged to 00 improve affinity or cross-reactivity. For example, aspartic acid at position 4 of the 9-mer core sequence is an optimal residue 0for DR3 binding, and substitution for that residue often improves DR 3 binding.

L) Example 17: Immunoaenlcitv of 109P1D4-derived HTL epitopes This example determines immunogenic DR supermotif- and DR3 motif-bearing epitopes among those identified Susing the methodology set forth herein.

Immunogenicity of HTL epitopes are confirmed in a manner analogous to the determination of immunogenicty of CTL epitopes, by assessing the ability to stimulate HTL responses and/or by using appropriate transgenic mouse models.

CK1 Immunogenidty is determined by screening for in vitro primary induction using normal PBMC or recall responses from Spatients who have 109P1D4-expressing tumors.

00 Example 18: Calculation of phenotypic frequencies of HLA-supertypes In various ethnic backgrounds to determine breadth of population coverage C1 This example illustrates the assessment of the breadth of population coverage of a vaccine composition comprised of multiple epitopes comprising multiple supermotifs and/or motifs.

In order to analyze population coverage, gene frequencies of HLA alleles are determined. Gene frequencies for each HLA allele are calculated from antigen or allele frequencies utilizing the binomial distribution formulae gf=1-lSQRT(1af)) (see, Sidney et al., Human Immunol. 45:79-93, 1996). To obtain overall phenotypic frequencies, cumulative gene frequencies are calculated, and the cumulative antigen frequencies derived by the use of the inverse formula [af=--(1-Cgf) 2 Where frequency data is not available at the level of DNA typing, correspondence to the serologically defined antigen frequencies is assumed. To obtain total potential supertype population coverage no linkage disequilibrium is assumed, and only alleles confirmed to belong to each of the supertypes are included (minimal estimates). Estimates of total potential coverage achieved by inter-lod combinations are made by adding to the A coverage the proportion of the non-A covered population that could be expected to be covered by the B alleles considered total=A+B*(1-A)). Confirmed members of the A3-like supertype are A3, All, A31, A*3301, and A*6801. Although the A3-like supertype may also include A34, A66, and A'7401, these alleles were not included in overall frequency calculations. Ukewise, confirmed members of the A2-lke supertype family are A*0201, A*0202, A*0203, A*0204, A*0205, A*0206, A*0207, A*6802, and A*6901. Finally, the 87-like supertype-confirmed alleles are: B7, B'3501-03, 851, B*5301, B*5401, B*5501-2, B*5601, 8*6701, and B7801 (potentially also B'1401, B*3504-06, B*4201, and B*5602).

Population coverage achieved by combining the A2-, A3- and B7-supertypes is approximately 86% in five major ethnic groups. Coverage may be extended by including peptides bearing the Al and A24 motifs. On average, Al is present in 12% and A24 in 29% of the population across five different major ethnic groups (Caucasian, North American Black, Chinese, Japanese, and Hispanic). Together, these alleles are represented with an average frequency of 39% in these same ethnic populations. The total coverage across the major ethnicities when Al and A24 are combined with the coverage of the A2-, A3- and B7-supertype alleles is see, Table IV An analogous approach can be used to estimate population coverage achieved with combinations of class II motif-bearing epitopes.

Immunogenicity studies in humans Bertoni et al., J. Clin. Invest. 100:503,1997; Doolan et al., Immunity 7:97, 1997; and Threlkeld et al., J. Immunol. 159:1648,1997) have shown that highly cross-reactive binding peptides are almost always recognized as epitopes. The use of highly cross-reactive binding peptides is an important selection criterion in identifying candidate epitopes for inclusion in a vaccine that is immunogenic in a diverse population.

With a sufficient number of epitopes (as disclosed herein and from the art), an average population coverage is 00 0 predicted to be greater than 95% in each of five major ethnic populations. The game theory Monte Carlo simulation analysis, Swhich is known in the art (see Osbome, M.J. and Rubinstein, A. "A course in game theory' MIT Press, 1994), can be Sused to estimate what percentage of the individuals in a population comprised of the Caucasian, North American Black, S Japanese, Chinese, and Hispanic ethnic groups would recognize the vaccine epitopes described herein. A preferred percentage is 90%. A more preferred percentage is Example 19: CTL Recoanition Of Endogenously Processed Antigens After Priming This example confirms that CTL induced by native or analoged peptide epitopes identified and selected as Sdescribed herein recognize endogenously synthesized, native antigens.

CK Effector cells isolated from transgenic mice that are immunized with peptide epitopes, for example HLA-A2 Ssupermotif-bearing epitopes, are re-stimulated in vitro using peptide-coated stimulator cells. Six days later, effector cells are 00 assayed for cytotoxicity and the cell lines that contain peptide-specific cytotoxic activity are further re-stimulated. An additional six days later, these cell lines are tested for cytotoxic activity on s 5 Cr labeled Jurkat-A2.1/Kb target cells in the absence or presence of peptide, and also tested on s 1 Cr labeled target cells bearing the endogenously synthesized antigen, i.e. cells that are stably transfected with 109P1D4 expression vectors.

The results demonstrate that CTL lines obtained from animals primed with peptide epitope recognize endogenously synthesized 109P1 D4 antigen. The choice of transgenic mouse model to be used for such an analysis depends upon the epitope(s) that are being evaluated. In addition to HLA-A*0201/K 1 transgenic mice, several other transgenic mouse models including mice with human All, which may also be used to evaluate A3 epitopes, and B7 alleles have been characterized and others transgenic mice for HLA-A1 and A24) are being developed. HLA-DR1 and HLA- DR3 mouse models have also been developed, which may be used to evaluate HTL epitopes.

Example 20: Activity Of CTL-HTL Conjuaated Epitopes In Transaenic Mice This example Illustrates the induction of CTLs and HTLs in transgenic mice, by use of a 109P1D4-derived CTL and HTL peptide vaccine compositions. The vaccine composition used herein comprise peptides to be administered to a patient with a 109P1D4-expressing tumor. The peptide composition can comprise multiple CTL and/or HTL epitopes. The epitopes are identified using methodology as described herein. This example also illustrates that enhanced immunogenicity can be achieved by inclusion of one or more HTL epitopes in a CTL vaccine composition; such a peptide composition can comprise an HTL epitope conjugated to a CTL epitope. The CTL epitope can be one that binds to multiple HLA family members at an affinity of 500 nM or less, or analogs of that epitope. The peptides may be lipidated, if desired.

Immunization procedures: Immunization of transgenic mice is performed as described (Alexander et al., J.

Immunol. 159:4753-4761, 1997). For example, A2/Kb mice, which are transgenic for the human HLA A21 allele and are used to confirm the immunogenicity of HLA-A*0201 motif- or HLA-A2 supermotif-bearing epitopes, and are primed subcutaneously (base of the tail) with a 0.1 ml of peptide in Incomplete Freund's Adjuvant, or If the peptide composition is a lipldated CTUHTL conjugate, in DMSO/saline, or if the peptide composition is a polypeptide, in PBS or Incomplete Freund's Adjuvant Seven days after priming, splenocytes obtained from these animals are restimulated with syngenic irradiated LPSactivated lymphoblasts coated with peptide.

Cell lines: Target cells for peptide-specific cytotoxicity assays are Jurkat cells transfected with the HLA-A2.1/Kb chimeric gepe Vitiello et al., J. Exp. Med. 173:1007,1991) Ijnvft CTL activation: One week after priming, spleen cells (30x106 cells/flask) are co-cultured at 37"C with syngeneic, irradiated (3000 rads), peptide coated lymphoblasts (10x10 6 cells/flask) In 10 ml of culture medium/T25 flask.

00 0 After six days, effector cells are harvested and assayed for cytotoxic activity.

L' Assay for cytotoxic activity: Target cells (1.0 to 1.5x10 6 are incubated at 37"C in the presence of 200 pl of 51 Cr.

L After 60 minutes, cells are washed three times and resuspended in R10 medium. Peptide is added where required at a concentration of 1 pgAnl. For the assay, 10 4 sCr-labeled target cells are added to different concentrations of effector cells S(final volume of 200 pl) in U-bottom 96-weU plates. After a six hour incubation period at 37"C, a 0.1 ml aliquot of Ssupematant is removed from each well and radioactivity is determined in a Micromedic automatic gamma counter. The percent specific lysis is determined by the formula: percent specific release 100 x (experimental release spontaneous release)/(maximum release spontaneous release). To facilitate comparison between separate CTL assays run under the same conditions, s 5 Cr release data is expressed as lytic unlts/106 cells. One lytic unit is arbitrarily defined as the number C of effector cells required to achieve 30% lysis of 10,000 target cells in a six hour 5 6 Cr release assay. To obtain specific lytic units/1 06, the lytic units/10 6 obtained in the absence of peptide is subtracted from the lytic units/10 6 obtained in the presence 00 of peptide. For example, if 30% 6sCr release is obtained at the effector target ratio of 50:1 5x10 5 effector cells 0for 10,000 targets) in the absence of peptide and 5:1 5x10 4 effector cells for 10,000 targets) in the presence of peptide, the specific lytic units would be: [(1/50,000)-(1/500,000)] x 106 18 LU.

The results are analyzed to assess the magnitude of the CTL responses of animals injected with the immunogenic CTUHTL conjugate vaccine preparation and are compared to the magnitude of the CTL response achieved using, for example, CTL epitopes as outlined above In the Example entitled 'Confirmation of Immunogenicity." Analyses similar to this may be performed to confirm the Immunogenicity of peptide conjugates containing multiple CTL epitopes and/or multiple HTL epitopes. In accordance with these procedures, it is found that a CTL response is induced, and concomitantly that an HTL response is induced upon administration of such compositions.

Example 21: Selection of CTL and HTL epitopes for Inclusion In a 109P1D4-speclfc vaccine.

This example illustrates a procedure for selecting peptide epitopes for vaccine compositions of the Invention. The peptides In the composition can be in the form of a nucleic acid sequence, either single or one or more sequences minigene) that encodes peptide(s), or can be single and/or polyepitopic peptides.

The following principles are utilized when selecting a plurality of epitopes for inclusion in a vaccine composition.

Each of the following principles is balanced in order to make the selection.

Epitopes are selected which, upon administration, mimic immune responses that are correlated with 109P1D4 clearance. The number of epitopes used depends on observations of patients who spontaneously cear 109P1D4. For example, If it has been observed that patients who spontaneously dear 109P D4-expressing cells generate an Immune response to at least three epitopes from 109P1D4 antigen, then at least three epitopes should be included for HLA class 1. A similar rationale is used to determine HLA dass II epitopes.

Epitopes are often selected that have a binding affinity of an ICso of 500 nM or less for an HLA class I molecule, or for class II, an ICso of 1000 nM or less; or HLA Class I peptides with high binding scores from the BIMAS web site, at URL bimas.dcrtnih.gov/.

In order to achieve broad coverage of the vaccine through out a diverse population, sufficient supermotif bearing peptides, or a sufficient array of allele-specific motif bearing peptides, are selected to give broad population coverage. In one embodiment, epitopes are selected to provide dt least 80% population coverage. A Monte Carlo analysis, a statistical evaluation known in the art, can be employed to assess breadth, or redundancy, of population coverage.

When creating polyepitoplc compositions, or a minigene that encodes same, it Is typically desirable to generate the smallest peptide possible that encompasses the epitopes of Interest The principles employed are similar, if not the same, as 00 0 those employed when selecting a peptide comprising nested epitopes. For example, a protein sequence for the vaccine Scomposition is selected because it has maximal number of epitopes contained within the sequence, it has a high concentration of epitopes. Epitopes may be nested or overlapping frame shifted relative to one another). For example, with overlapping epitopes, two 9-mer epitopes and one 10-mer epitope can be present in a 10 amino acid peptide. Each epitope can be exposed and bound by an HLA molecule upon administration of such a peptide. A multi-epitopic, peptide can be generated synthetically, recombinantly, or via cleavage from the native source. Alternatively, an analog can be made of this native sequence, whereby one or more of the epitopes comprise substitutions that alter the cross-reactivity and/or Sbinding affinity properties of the polyepitopic peptide. Such a vaccine composition is administered for therapeutic or prophylactic purposes. This embodiment provides for the possibility that an as yet undiscovered aspect of immune system C processing will apply to the native nested sequence and thereby facilitate the production of therapeutic or prophylactic immune response-inducing vaccine compositions. Additionally such an embodiment provides for the possibility of motif- 00 bearing epitopes for an HLA makeup that is presently unknown. Furthermore, this embodiment (absent the creating of any F analogs) directs the immune response to multiple peptide sequences that are actually present in 109P1D4, thus avoiding the C need to evaluate any junctional epitopes. Lastly, the embodiment provides an economy of scale when producing nucleic add vaccine compositions. Related to this embodiment computer programs can be derived in accordance with principles in the art, which identify in a target sequence, the greatest number of epitopes per sequence length.

A vaccine composition comprised of selected peptides, when administered, is safe, efficacious, and elicits an immune response similar in magnitude to an immune response that controls or clears cells that bear or overexpress 109P1D4.

Example 22: Construction of "Minlaene" Muti-Epitope DNA Plasmids This example discusses the construction of a minigene expression plasmid. Minigene plasmids may, of course, contain various configurations of B cell, CTL and/or HTL epitopes or epitope analogs as described herein.

A minigene expression plasmid typically includes multiple CTL and HTL peptide epitopes. In the present example, HLA-A2, -A3, -B7 supermotif-bearing peptide epitopes and HLA-A1 and -A24 motif-bearing peptide epitopes are used in conjunction with DR supermotif-bearing epitopes and/or DR3 epitopes. HLA class I supermotif or motif-bearing peptide epitopes derived 109P1D4, are selected such that multiple supermotifs/motifs are represented to ensure broad population coverage. Similarly, HLA class II epitopes are selected from 109P1 D4 to provide broad population coverage, I.e. both HLA DR-1-4-7 supermotif-bearing epitopes and HLA DR-3 motif-bearing epitopes are selected for inclusion in the minigene construct The selected CTL and HTL epitopes are then incorporated into a minigene for expression in an expression vector.

Such a construct may additionally include sequences that direct the HTL epitopes to the endoplasmic reticulum.

For example, the li protein may be fused to one or more HTL epitopes as described in the art, wherein the CLIP sequence of the li protein is removed and replaced with an HLA dass II epitope sequence so that HLA class II epitope Is directed to the endoplasmic reticulum, where the epitope binds to an HLA dass II molecules.

This example illustrates the methods to be used for construction of a minlgene-bearing expression plasmid. Other expression vectors that may be used for minigene compositions are available and known to those of skill in the art.

The minigene DNA plasmid of this example contains a consensus Kozak sequence and a consensus murine kappa Ig-light chain signal sequence followed by CTL and/or HTL epitopes selected in accordance with principles disclosed herein. The sequence encodes an open reading frame fused to the Myc and His antibody epitope tag coded for by the pcDNA 3.1 Myc-His vector.

Overlapping oligonucleotides that can, for example, average about 70 nucleotides in length with 15 nucleotide 0 overlaps, are synthesized and HPLC-purified. The oligonudeotides encode the selected peptide epitopes as well as appropriate linker nudeotides, Kozak sequence, and signal sequence. The final multiepitope minigene is assembled by extending the overlapping oligonudeotides in three sets of reactions using PCR. A Perkin/Elmer 9600 PCR machine is used and a total of 30 cycles are performed using the following conditions: 950C for 15 sec, annealing temperature below the lowest calculated Tm of each primer pair) for 30 sec, and 72C0 for 1 min.

For example, a minigene is prepared as follows. For a first PCR reaction, 5 pg of each of two oligonudeotides are annealed and extended: In an example using eight oligonucleotides, four pairs of primers, oligonudeotides 1+2, 3+4, 5+6, and 7+8 are combined in 100 pl reactions containing Pfu polymerase buffer (1x= 10 mM KCL, 10 mM (NH4)2SO4, C mM Tris-chloride, pH 8.75, 2 mM MgSO4, 0.1% Triton X-100, 100 pg/ml BSA), 0.25 mM each dNTP, and 2.5 U of Pfu CK polymerase. The full-length dimer products are gel-purified, and two reactions containing the product of 1+2 and 3+4, and Sthe product of 5+6 and 7+8 are mixed, annealed, and extended for 10 cycles. Half of the two reactions are then mixed, and 00 5 cycles of annealing and extension carded out before flanking primers are added to amplify the full length product The fulllength product is gel-purified and cloned into pCR-blunt (Invitrogen) and individual clones are screened by sequencing.

Example 23: The Plasmid Construct and the Degree to Which It Induces Immunoaenlcty.

The degree to which a plasmid construct, for example a plasmid constructed in accordance with the previous Example, is able to induce immunogenicity is confirmed in vitro by determining epitope presentation by APC following transduction or transfection of the APC with an epitope-expressing nucleic acid construct Such a study determines 'antigenidty" and allows the use of human APC. The assay determines the ability of the epitope to be presented by the APC in a context that is recognized by a T cell by quantifying the density of epitope-HLA class I complexes on the cell surface.

Quantitation can be performed by directly measuring the amount of peptide eluted from the APC (see, Sljts et al., J.

Immunol. 156:683-692, 1996; Demotz et al., Nature 342:682-684, 1989); or the number of peptide-HLA class I complexes can be estimated by measuring the amount of lysis or lymphokine release induced by diseased or transfected target cells, and then determining the concentration of peptide necessary to obtain equivalent levels of lysis or lymphokine release (see, Kageyama et J. Immunol. 154:567-576, 1995).

Altematively, immunogenicity is confirmed through in vivo injections into mice and subsequent in vitro assessment of CTL and HTL activity, which are analyzed using cytotoxicity and proliferation assays, respectively, as detailed in Alexander et al., Immunity 1:751-761, 1994.

For example, to confirm the capacity of a DNA minigene construct containing at least one HLA-A2 supermotif peptide to induce CTLs in vivo, HLA-A2.1/K b transgenic mice, for example, are immunized intramuscularly with 100 ipg of naked cDNA. As a means of comparing the level of CTLs induced by cDNA immunization, a control group of animals is also immunized with an actual peptide composition that comprises multiple epitopes synthesized as a single polypeptide as they would be encoded by the minlgene.

Splenocytes from immunized animals are stimulated twice with each of the respective compositions (peptide epitopes encoded in the minigene or the polyepitopic peptide), then assayed for peptide-specific cytotoxic activity In a slCr release assay. The results indicate the magnitude of the CTL response directed against the A2-restricted epitope, thus indicating the In vivo immunogenicity of the minigene vaccine and polyepitopic vaccine.

It is, therefore, found that the minlgene elicits immune responses directed toward the HLA-A2 supermotif peptide epitopes as does the polyepitopic peptide vaccine. A similar analysis is also performed using other HLA-A3 and HLA-B7 transgenic mouse models to assess CTL induction by HLA-A3 and HLA-B7 motif or supermotif epitopes, whereby It is also found that the minigene elicits appropriate immune responses directed toward the provided epitopes.

00 0 To confirm the capacity of a class II epitope-encoding minigene to induce HTLs in vivo, DR transgenic mice, or for those epitopes that cross react with the appropriate mouse MHC molecule, I-Abrestricted mice, for example, are immunized a intramuscularly with 100 pg of plasmid DNA. As a means of comparing the level of HTLs induced by DNA immunization, a group of control animals is also Immunized with an actual peptide composition emulsified in complete Freund's adjuvant Cj) CD4+ T cells, l.e. HTLs, are purified from splenocytes of immunized animals and stimulated with each of the respective 0 compositions (peptides encoded in the minigene). The HTL response is measured using a 3 H-thymidine incorporation proliferation assay, (see, Alexander et a. Immunity 1:751-761, 1994). The results indicate the magnitude of the HTL response, thus demonstrating the in vivo immunogenidty of the minigene.

SDNA minigenes, constructed as described in the previous Example, can also be confirmed as a vaccine in N combination with a boosting agent using a prime boost protocol. The boosting agent can consist of recombinant protein Bamett et al., Aids Res. and Human Retroviruses 14, Supplement 3:S299-S309, 1998) or recombinant vaccinia, for 00 example, expressing a minigene or DNA encoding the complete protein of interest (see, Hanke et Vaccine 16:439- 445, 1998; Sedegah et al., Proc. Natl. Acad. Sl USA 95:7648-53, 1998; Hanke and McMichael, Immunol. Letters 66:177- S181, 1999; and Robinson et al., Nature Med. 5:526-34, 1999).

For example, the efficacy of the DNA minigene used in a prime boost protocol is initially evaluated in transgenic mice. In this example, A21/Kb transgenic mice are immunized IM with 100 ig of a DNA minigene encoding the immunogenic peptides including at least one HLA-A2 supermotif-bearing peptide. After an incubation period (ranging from 3- 9 weeks), the mice are boosted IP with 107 pfulmouse of a recombinant vaccinia virus expressing the same sequence encoded by the DNA minigene. Control mice are immunized with 100 pg of DNA or recombinant vaccinia without the minigene sequence, or with DNA encoding the minigene, but without the vaccnia boost After an additional incubation period of two weeks, splenocytes from the mice are immediately assayed for peptide-specific activity in an ELISPOT assay.

Additionally, splenocytes are stimulated In vitro with the A2-restricted peptide epitopes encoded in the minigene and recombinant vaccinia, then assayed for peptide-specific activity in an alpha, beta and/or gamma IFN EUSA.

It is found that the minigene utilized in a prime-boost protocol elicits greater Immune responses toward the HLA-A2 supermotif peptides than with DNA alone. Such an analysis can also be performed using HLA-A11 or HLA-B7 transgenic mouse models to assess CTL induction by HLA-A3 or HLA-B7 motif or supermotif epitopes. The use of prime boost protocols in humans is described below in the Example entitled 'Induction of CTL Responses Using a Prime Boost Protocol." Example 24: Peptide Compositions for Prophylactic Uses Vaccine compositions of the present invention can be used to prevent 109P1D4 expression in persons who are at risk for tumors that bear this antigen. For example, a polyepitopic peptide epitope composition (or a nucleic acid comprising the same) containing multiple CTL and HTL epitopes such as those selected In the above Examples, which are also selected to target greater than 80% of the population, is administered to individuals at risk for a 109P1D4-associated tumor.

For example, a peptide-based composition is provided as a single polypeptide that encompasses multiple epitopes. The vaccine Is typically administered in a physiological solution that comprises an adjuvant, such as Incomplete Freunds Adjuvant The dose of peptide for the initial immunization is from about 1 to about 50,000 pg, generally 100-5,000 pg, for a 70 kg patient The Initial administration of vaccine is followed by booster dosages at 4 weeks followed by evaluation of the magnitude of the immune response in the patient, by techniques that determine the presence of epitopespecific CTL populations in a PBMC sample. Additional booster doses are administered as required. The composition Is found to be both safe and efficacious as a prophylaxis against 109P1D4-assocdated disease.

Altematively, a composition typically comprising transfecting agents is used for the administration of a nucleic add- 109 00 0 based vaccine in accordance with methodologies known in the art and disclosed herein.

Example 25: Polvepitoplc Vaccine Compositions Derived from Native 109P1D4 Sequences L) A native 109P1D4 polyprotein sequence is analyzed, preferably using computer algorithms defined for each class I and/or class II supermoif or motif, to identify 'relatively short' regions of the polyprotein that comprise multiple epitopes. The "relatively short regions are preferably less in length than an entire native antigen. This relatively short sequence that contains multiple distinct or overlapping, 'nested" epitopes can be used to generate a minigene construct The construct is Sengineered to express the peptide, which corresponds to the native protein sequence. The "relatively short" peptide is C generally less than 250 amino acids in length, often less than 100 amino acids in length, preferably less than 75 amino acids Sin length, and more preferably less than 50 amino acids in length. The protein sequence of the vaccine composition is selected because it has maximal number of epitopes contained within the sequence, it has a high concentration of 00 epitopes. As noted herein, epitope motifs may be nested or overlapping frame shifted relative to one another). For example, with overlapping epitopes, two 9-mer epitopes and one 10-mer epitope can be present in a 10 amino acid peptide.

CN Such a vaccine composition Is administered for therapeutic or prophylactic purposes.

The vaccine composition will include, for example, multiple CTL epitopes from 109P1D4 antigen and at least one HTL epitope. This polyepitopic native sequence is administered either as a peptide or as a nucleic add sequence which encodes the peptide. Alternatively, an analog can be made of this native sequence, whereby one or more of the epitopes comprise substitutions that alter the cross.reactivity and/or binding affinity properties of the polyepitopic peptide.

The embodiment of this example provides for the possibility that an as yet undiscovered aspect of immune system processing will apply to the native nested sequence and thereby facilitate the production of therapeutic or prophylactic immune response-inducing vaccine compositions. Additionally, such an embodiment provides for the possibility of motifbearing epitopes for an HLA makeup(s) that is presently unknown. Furthermore, this embodiment (excluding an analoged embodiment) directs the immune response to multiple peptide sequences that are actually present in native 109P1D4, thus avoiding the need to evaluate any junctional epitopes. Lastly, the embodiment provides an economy of scale when producing peptide or nucleic acd vaccine compositions.

Related to this embodiment, computer programs are available in the art which can be used to identify in a target sequence, the greatest number of epitopes per sequence length.

Example 26: Polvepitopic Vaccine Compositions from Multiple Antiaens The 109P1 D4 peptide epitopes of the present invention are used in conjunction with epitopes from other target tumor-associated antigens, to create a vaccine composition that is useful for the prevention or treatment of cancer that expresses 109P1D4 and such other antigens. For example, a vaccine composition can be provided as a single polypeptide that incorporates multiple epitopes from 109P1D4 as well as tumor-associated antigens that are often expressed with a target cancer associated with 109P1D4 expression, or can be administered as a composition comprising a cocktail of one or more discrete epitopes. Altematively, the vaccine can be administered as a minigene construct or as dendritic cells which have been loaded with the peptide epitopes in vitro.

Example 27: Use of peptides to evaluate an immune response Peptides of the invention may be used to analyze an immune response for the presence of specific antibodies, CTL or HTL directed to 109P1D4. Such an analysis can be performed in a manner described by Ogg et el., Science 279:2103-2106,1998. In this Example, peptides in accordance with the invention are used as a reagent for diagnostic or 00 0 prognostic purposes, not as an immunogen.

SIn this example highly sensitive human leukocyte antigen tetrameric complexes (tetramers) are used for a cross- Ssectional analysis of, for example, 109P1D4 HLA-A'0201-specific CTL frequencies from HLA A*0201-positive Individuals at L) different stages of disease or following immunization comprising a 109P1D4 peptide containing an A*0201 motif. Tetrameric complexes are synthesized as described (Musey et al., N. Engl. J. Med. 337:1267, 1997). Briefly, purified HLA heavy chain O (A*0201 in this example) and p2-microglobulin are synthesized by means of a prokaryotic expression system. The heavy chain is modified by deletion of the transmembrane-cytosolic tail and COOH-terminal addition of a sequence containing a 0BirA enzymatic biotinylation site. The heavy chain, p2-microglobulin, and peptide are refolded by dilution. The refolded product is isolated by fast protein liquid chromatography and then blotinylated by BirA in the presence of biotin CN (Sigma, St Louis, Missouri), adenosine 5' triphosphate and magnesium. Streptavidin-phycoerythrin conjugate is added in a C 1:4 molar ratio, and the teframeric product is concentrated to 1 mg/ml. The resulting product is referred to as tetramer- 00 phycoerythrin.

SFor the analysis of patient blood samples, approximately one million PBMCs are centrifuged at 300g for 5 minutes and resuspended in 50 0p of cold phosphate-buffered saline. Tri-color analysis is performed with the tetramer-phycoerythrin, along with anti-CD8-Tricolor, and anti-CD38. The PBMCs are incubated with tetramer and antibodies on ice for 30 to 60 min and then washed twice before formaldehyde fixation. Gates are applied to contain >99.98% of control samples. Controls for the tetramers include both A*0201-negative individuals and A'0201-positive non-diseased donors. The percentage of cells stained with the tetramer is then determined by flow cytometry. The results indicate the number of cells in the PBMC sample that contain epitope-restricted CTLs, thereby readily indicating the extent of immune response to the 109P1D4 epitope, and thus the status of exposure to 109P1D4, or exposure to a vaccine that elicits a protective or therapeutic response.

Example 28: Use of Peptide Epitopes to Evaluate Recall Responses The peptide epitopes of the invention are used as reagents to evaluate T cell responses, such as acute or recall responses, in patients. Such an analysis may be performed on patients who have recovered from 109P1D4-associated disease or who have been vaccinated with a 109P1D4 vaccine.

For example, the class I restricted CTL response of persons who have been vaccinated may be analyzed. The vaccine may be any 109P1 D4 vaccine. PBMC are collected from vaccinated individuals and HLA typed. Appropriate peptide epitopes of the invention that, optimally, bear supermotifs to provide cross-reactivity with multiple HLA supertype family members, are then used for analysis of samples derived from individuals who bear that HLA type.

PBMC from vaccinated individuals are separated on Ficoll-Histopaque density gradients (Sigma Chemical Co., St Louis, MO), washed three times in HBSS (GIBCO Laboratories), resuspended in RPMI-1640 (GIBCO Laboratories) supplemented with L-glutamine (2mM), penicillin (50U/ml), streptomycn (50 pg/ml), and Hepes (10mM) containing heat-inactivated human AB serum (complete RPMI) and plated using microculture formats. A synthetic peptide comprising an epitope of the invention is added at 10 pg/ml to each well and HBV core 128-140 epitope is added at 1 pg/ml to each well as a source of T cell help during the first week of stimulation.

In the microculture format, 4 x 105 PBMC are stimulated with peptide in 8 replicate cultures in 96-well round bottom plate in 100 pi/well of complete RPMI. On days 3 and 10, 100 pl of complete RPMI and 20 U/ml final concentration of rlL-2 are added to each well. On day 7 the cultures are transferred into a 96-well flat-bottom plate and restimulated with peptide, rlL-2 and 105 irradiated (3,000 rad) autologous feeder cells. The cultures are tested for cytotoxic activity on day 14. A positive CTL response requires two or more of the eight replicate cultures to display greater than 10% specific 5 Cr release, based on comparison with non-diseased control subjects as previously described (Rehermann, et Nature Med.

00 0 2:1104,1108, 1996; Rehermann et al., J. Clin. Invest. 97:1655-1665, 1996; and Rehermann et a. J. Clin. Invest. 98:1432- S1440, 1996).

Q_ Target cell lines are autologous and allogenelc EBV-transformed B-LCL that are either purchased from the SAmerican Society for Histocompatiblity and Immunogenetics (ASHI, Boston, MA) or established from the pool of patients as described (Guilhot, etal. J. Viol. 66:2670-2678, 1992).

SCytotoxicity assays are performed in the following manner. Target cells consist of either allogenelc HLA-matched or autologous EBV-transformed B lymphoblastoid cell line that are incubated ovemight with the synthetic peptide epitope of the Invention at 10 pM, and labeled with 100 tpCi of 5 'Cr (Amersham Corp., Arlington Heights, IL) for 1 hour after which they are washed four times with HBSS.

CK Cytolytic activity is determined in a standard 4-h, split well 5 Cr release assay using U-bottomed 96 well plates containing 3,000 targets/well. Stimulated PBMC are tested at effector/target ratios of 20-501 on day 14. Percent 00 cytotoxlcity is determined from the formula: 100 x [(experimental release-spontaneous release)/maximum release- 0spontaneous release)j. Maximum release is determined by lysis of targets by detergent Triton X-100; Sigma Chemical Co., St Louis, MO). Spontaneous release is <25% of maximum release for all experiments.

The results of such an analysis Indicate the extent to which HLA-restricted CTL populations have been stimulated by previous exposure to 109P1D4 or a 109P1D4 vaccine.

Similarly, Class II restricted HTL responses may also be analyzed. Purified PBMC are cultured in a 96-well flat bottom plate at a density of 1.5x10 5 cells/well and are stimulated with 10 pg/ml synthetic peptide of the invention, whole 109P1D4 antigen, or PHA. Cells are routinely plated in replicates of 4-6 wells for each condition. After seven days of culture, the medium is removed and replaced with fresh medium containing 10U/ml IL-2. Two days later, 1 pCi 3H-thymidine is added to each well and incubation is continued for an additional 18 hours. Cellular DNA is then harvested on glass fiber mats and analyzed for 3 H-thymidine incorporation. Antigen-specific T cell proliferation is calculated as the ratio of 3Hthymidine incorporation In the presence of antigen divided by the 3 H-thymidine incorporation In the absence of antigen.

Example 29: Induction Of Specific CTL Response In Humans A human clinical trial for an immunogenic composition comprising CTL and HTL epitopes of the Invention is set up as an IND Phase I, dose escalation study and carried out as a randomized, double-blind, placebo-controlled trial. Such a trial is designed, for example, as follows: A total of about 27 Individuals are enrolled and divided into 3 groups: Group I: 3 subjects are injected with placebo and 6 subjects are injected with 5 pg of peptide composition; Group II: 3 subjects are injected with placebo and 6 subjects are Injected with 50 pg peptide composition; Group II: 3 subjects are injected with placebo and 6 subjects are injected with 500 pg of peptide composition.

After 4 weeks following the first injection, all subjects receive a booster inoculation at the same dosage.

The endpoints measured In this study relate to the safety and tolerability of the peptide composition as well as Its immunogenlcity. Cellular immune responses to the peptide composition are an index of the intrinsic activity of this the peptide composition, and can therefore be viewed as a measure of biological efficacy. The following summarize the clinical and laboratory data that relate to safety and efficacy endpoints.

Safety. The incidence of adverse events Is monitored in the placebo and drug treatment group and assessed in terms of degree and reversibility.

Evaluation of Vaccine Efficacy: For evaluation of vaccine efficacy, subjects are bled before and after injection.

Peripheral blood mononuclear cells are Isolated from fresh heparinized blood by Ficoll-Hypaque density gradient Scentrifugation, aliquoted in freezing media and stored frozen. Samples are assayed for CTL and HTL activity.

The vaccine is found to be both safe and efficacious.

Example 30: Phase If Trials In Patients Expressing 109P1D4 Phase II trials are performed to study the effect of administering the CTL-HTL peptide compositions to patients Shaving cancer that expresses 109P1D4. The main objectives of the trial are to determine an effective dose and regimen for inducing CTLs in cancer patients that express 109P1D4, to establish the safety of inducing a CTL and HTL response in these patients, and to see to what extent activation of CTLs improves the clinical picture of these patients, as manifested, Se.g., by the reduction and/or shrinking of lesions. Such a study is designed, for example, as follows: CThe studies are performed in multiple centers. The trial design is an open-label, uncontrolled, dose escalation Sprotocol wherein the peptide composition is administered as a single dose followed six weeks later by a single booster shot 00 of the same dose. The dosages are 50, 500 and 5,000 micrograms per injection. Drug-associated adverse effects (severity and reversibility) are recorded.

There are three patient groupings. The first group is injected with 50 micrograms of the peptide composition and the second and third groups with 500 and 5,000 micrograms of peptide composition, respectively. The patients within each group range in age from 21-65 and represent diverse ethnic backgrounds. All of them have a tumor that expresses 109P1D4.

Clinical manifestations or antigen-specific T-cell responses are monitored to assess the effects of administering the peptide compositions. The vaccine composition is found to be both safe and efficacious in the treatment of 109P1D4associated disease.

Example 31: Induction of CTL Responses Using a Prime Boost Protocol A prime boost protocol similar in its underlying principle to that used to confirm the efficacy of a DNA vaccine in transgenic mice, such as described above in the Example entitled "The Plasmid Construct and the Degree to Which It Induces Immunogenicity,' can also be used for the administration of the vaccine to humans. Such a vaccine regimen can include an initial administration of, for example, naked DNA followed by a boost using recombinant virus encoding the vaccine, or recombinant protein/polypeptide or a peptide mixture administered in an adjuvant For example, the initial immunization may be performed using an expression vector, such as that constructed in the Example entitled "Construction of Minigene" Multi-Epitope DNA Plasmlds" in the form of naked nucleic acid administered IM (or SC or ID) in the amounts of 0.5-5 mg at multiple sites. The nucleic acid (0.1 to 1000 pg) can also be administered using a gene gun. Following an incubation period of 3-4 weeks, a booster dose Is then administered. The booster can be recombinant fowlpox virus administered at a dose of 5-10 7 to 5x109 pfu. An altemative recombinant virus, such as an MVA, canarypox, adenovirus, or adeno-associated virus, can also be used for the booster, or the polyepitopic protein or a mixture of the peptides can be administered. For evaluation of vaccine efficacy, patient blood samples are obtained before immunization as well as at intervals following administration of the initial vaccine and booster doses of the vaccine.

Peripheral blood mononudear cells are isolated from fresh heparinized blood by Ficoll-Hypaque density gradient centrifugation, aliquoted in freezing media and stored frozen. Samples are assayed for CTL and HTL activity.

Analysis of the results indicates that a magnitude of response sufficient to achieve a therapeutic or protective immunity against 109P1D4 is generated.

Example 32: Administration of Vaccine Compositions Using Dendritic Cells (DC) 00 0Vaccines comprising peptide epitopes of the invention can be administered using APCs, or "professional" APCs Ssuch as DC. In this example, peptide-pulsed DC are administered to a patient to stimulate a CTL response in vivo. In this Smethod, dendritic cells are isolated, expanded, and pulsed with a vaccine comprising peptide CTL and HTL epitopes of the invention. The dendritic cells are infused back into the patient to elicit CTL and HTL responses in vivo. The induced CTL and HTL then destroy or facilitate destruction, respectively, of the target cells that bear the 109P1D4 protein from which the epitopes in the vaccine are derived.

For example, a cocktail of epitope-comprising peptides is administered ex vivo to PBMC, or isolated DC therefrom.

A pharmaceutical to facilitate harvesting of DC can be used, such as ProgenipoietinM (Monsanto, St. Louis, MO) or GM- C N CSF/IL-4. After pulsing the DC with peptides, and prior to reinfusion into patients, the DC are washed to remove unbound S peptides.

SAs appreciated clinically, and readily determined by one of skill based on clinical outcomes, the number of DC 00 relnfused Into the patient can vary (see, Nature Med. 4:328, 1998; Nature Med. 2:52, 1996 and Prostate 32:272, 1997).

Although 2-50 x 106 DC per patient are typically administered, larger number of DC, such as 107 or 108 can also be provided.

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Such cell populations typically contain between 50-90% DC.

In some embodiments, peptide-loaded PBMC are injected into patients without purification of the DC. For example, PBMC generated after treatment with an agent such as Progenipoletin' M are injected into patients without purification of the DC. The total number of PBMC that are administered often ranges from 108 to 1010. Generally, the cell doses injected into patients is based on the percentage of DC in the blood of each patient, as determined, for example, by immunofluorescence analysis with specific anti-DC antibodies. Thus, for example, if Progenipoletin T mobilizes 2% DC in the peripheral blood of a given patient, and that patient is to receive 5 x 106 DC, then the patient will be injected with a total of 2.5 x 108 peptide-loaded PBMC. The percent DC mobilized by an agent such as Progenipoletin T M is typically estimated to be between 2-10%, but can vary as appreciated by one of skill in the art.

Ex vive activation of CTUHTL responses Alternatively, ex vivo CTL or HTL responses to 109P1D4 antigens can be induced by incubating, in tissue culture, the patient's, or genetically compatible, CTL or HTL precursor cells together with a source of APC, such as DC, and immunogenic peptides. After an appropriate incubation time (typically about 7-28 days), In which the precursor cells are activated and expanded Into effector cells, the cells are Infused into the patient, where they will destroy (CTL) or facilitate destruction (HTL) of their specific target cells, tumor cells.

Example 33: An Alternative Method of Identifying and Confirming Motif-Bearing Peptides Another method of Identifying and confirming motif-bearing peptides Is to elute them from cells bearing defined MHC molecules. For example, EBV transformed B cell lines used for tissue typing have been extensively characterized to determine which HLA molecules they express. In certain cases these cells express only a single type of HIA molecule.

These cells can be transfected with nudeic acids that express the antigen of interest, e.g. 109P1D4. Peptides produced by endogenous antigen processing of peptides produced as a result of transfection will then bind to HLA molecules within the cell and be transported and displayed on the cell's surface. Peptides are then eluted from the HLA molecules by exposure to mild add conditions and their amino acid sequence determined, by mass spectral analysis Kubo et al., J.

Immunol. 152:3913, 1994). Because the majority of peptides that bind a particular HLA molecule are motif-bearing, this is an alternative modality for obtaining the motif-bearing peptides correlated with the particular HLA molecule expressed on the cell.

Alteratively, cell lines that do not express endogenous HLA molecules can be transfected with an expression 00 0 construct encoding a single HLA allele. These cells can then be used as described, they can then be transfected with nucleic acids that encode 109P1D4 to isolate peptides corresponding to 109P1D4 that have been presented on the cell surface. Peptides obtained from such an analysis will bear motif(s) that correspond to binding to the single HLA allele that is S expressed in the cell.

As appreciated by one in the art, one can perform a similar analysis on a cell bearing more than one HLA allele 0and subsequently determine peptides specific for each HLA allele expressed. Moreover, one of skill would also recognize that means other than transfection, such as loading with a protein antigen, can be used to provide a source of antigen to the Scell.

CK Example 34: Complementary Polvnudcleotides ,Sequences complementary to the 109P1D4-encoding sequences, or any parts thereof, are used to detect, 00 decrease, or Inhibit expression of naturally occurring 109P1D4. Although use of oligonudeotides comprising from about Sto 30 base pairs is described, essentially the same procedure s used with smaller or with larger sequence fragments.

l Appropriate oligonuceotides are designed using, OLIGO 4.06 software (National Biosdcences) and the coding sequence of 109P1D4. To inhibit transcription, a complementary oligonucleotide is designed from the most unique 5' sequence and used to prevent promoter binding to the coding sequence. To inhibit translation, a complementary oligonudcleotbde is designed to prevent ribosomal binding to a 109P1D4-encoding transcript.

Example 35: Purification of Naturally-occurring or Recombinant 109P1D4 Uslng 109P1D4-Spedfic Antibodies Naturally occurring or recombinant 109P1 D4 is substantially purified by Immunoaffinity chromatography using antibodies specific for 109P1D4. An Immunoaffinity column is constructed by covalently coupling anti-109P1 D4 antibody to an activated chromatographic resin, such as CNBr-activated SEPHAROSE (Amersham Pharmadca Biotech). After the coupling, the resin is blocked and washed according to the manufacturer's instructions.

Media containing 109P1 D4 are passed over the immunoaffinity column, and the column is washed under conditions that allow the preferential absorbance of 1 09P1D4 high ionic strength buffers in the presence of detergent).

The column is eluted under conditions that disrupt antibody/109P1D4 binding a buffer ofpH 2 to pH 3, or a high concentration of a chaotrope, such as urea or thiocyanate ion), and GCR.P is collected.

Example 36: Identification of Molecules Which Interact with 109P1D4 109P1D4, or biologically active fragments thereof, are labeled with 1211 Bolton-Hunter reagent (See, Bolton et al. (1973) Biochem. J. 133:529.) Candidate molecules previously arrayed in the wells of a multi-well plate are incubated with the labeled 109P1D4, washed, and any wells with labeled 109P1D4 complex are assayed. Data obtained using different concentrations of 109P1D4 are used to calculate values for the number, affinity, and association of 109P1D4 with the candidate molecules.

Exampe 37: In Vivo Assay for 109P1D4 Tumor Growth Promotion The effect of a 109P1D4 protein on tumor cell growth is evaluated in vivo by gene overexpresslon In tumor-bearing mice. For example, SCID mice are Injected subcutaneously on each flank with 1 x 106 of either PC3, DU145 or 3T3 cells containing tkNeo empty vector or a nucleic acid sequence of the invention. At least two strategies can be used: (1) Constitutive expression under regulation of a promoter such as a constitutive promoter obtained from the genomes of viruses such as polyoma virus, fowIpox virus (UK 2,211,504 published 5 July 1989), adenovirus (such as Adenovirus bovine

I

0 papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and Simian Virus 40 (SV40), or from 0 heterologous mammalian promoters, the actin promoter or an immunoglobulin promoter, provided such promoters are compatible with the host cell systems, and Regulated expression under control of an Inducible vector system, such as ecdysone, tet, etc., provided such promoters are compatible with the host cell systems. Tumor volume is then monitored at the appearance of palpable tumors and followed over time to determine if the cells expressing a gene of the invention grow at a faster rate and whether tumors of a 109P1 D4 protein-expressing cells demonstrate characteristics of altered aggressiveness enhanced metastasis, vascularization, reduced responsiveness to chemotherapeutic drugs).

Additionally, mice can be implanted with 1 x 10 5 of the same cells orthotopically to determine if a protein of the invention has an effect on local growth in the prostate or on the ability of the cells to metastasize, specifically to lungs, lymph nodes, and bone marrow.

The assay is also useful to determine the inhibitory effect of candidate therapeutic compositions, such as for OO example, 109P1D4 protein-related intrabodies, 109P1D4 gene-related antisense molecules and ribozymes.

0 NC Example 38: 109P1D4 MonoclonalAntibody-mediated Inhibitton of Tumors In Vvo The significant expression of 109P1D4 proteins in the cancer tissues of Table I and its restrictive expression in normal tissues, together with its expected cell surface expression, makes 109P1D4 proteins excellent targets for antibody therapy. Similarly, 109P1D4 proteins are a target for T cell-based Immunotherapy. Thus, for 109P1D4 genes expressed, in prostate cancer, the therapeutic efficacy of anti-109P1 D4 protein mAbs in human prostate cancer xenograft mouse models is evaluated by using androgen-independent LAPC-4 and LAPC-9 xenografts (Craft, et Cancer Res, 1999.

59(19): p. 5030-6) and the androgen independent recombinant cell line PC3-of 109P1D4 (see, Kalghn, etal., Invest Urol, 1979. 17(1): p. 16-23); analogous models are used for other cancers.

Antibody efficacy on tumor growth and metastasis formation is studied, in a mouse orthotopic prostate cancer xenograft models and mouse kidney xenograft models. The antibodies can be unconjugated, as discussed in this Example, or can be conjugated to a therapeutic modality, as appreciated in the art Anti-109P1 D4 protein mAbs inhibit formation of both the androgen-dependent LAPC-9 and androgen-independent PC3-109P1 D4 protein tumor xenografts. Anti-109P1D4 protein mAbs also retard the growth of established orthotopic tumors and prolonged survival of tumor-bearing mice. These results indicate the utility of anti-109P1D4 protein mAbs in the treatment of local and advanced stages of prostate cancer.

(See, (Saffran, et al., PNAS 10:1073-1078 or World Wide Web URL ww.pnas.org/cgi/doll/0.1073/pnas.051624698).

Administration of the anti-109P1D4 protein mAbs lead to retardation of established orthotopic tumor growth and Inhibition of metastasis to distant sites, resulting in a significant prolongation in the survival of tumor-bearing mice. These studies indicate that proteins of the invention are attractive targets for immunotherapy and demonstrate the therapeutic potential of anti-109P1D4 protein mAbs for the treatment of local and metastatic cancer. This example demonstrates that unconjugated 109P1D4 protein-related monoclonal antibodies are effective to inhibit the growth of human prostate tumor xenografts and human kidney xenografts grown in SCID mice; accordingly a combination of such efficacious monoclonal antibodies is also effective.

Tumor inhibition using multiple unconjugated mAbs Materials and Methods 109P1D4 Protein-related Monodonal Antibodies: Monoclonal antibodies are raised against proteins of the invention as described in the Example entitled "Generation of 109P1D4 Monoclonal Antibodies". The antibodies are characterized by ELISA, Western blot, FACS, and 1 00 immunoprecipitation for their capacity to bind to the respective protein of the invention. Epitope mapping data for, the Santi-109P1D4 protein mAbs, as determined by ELISA and Western analysis, indicate that the antibodies recognize epitopes on the respective 109P1 D4 protein. Immunohistochemical analysis of prostate cancer tissues and cells with these S antibodies is performed.

Cj) The monoclonal antibodies are purified from ascites or hybridoma tissue culture supematants by Protein-G SSepharose chromatography, dialyzed against PBS, filter sterilized, and stored at -20°C. Protein determinations are performed by a Bradford assay (Bio-Rad, Hercules, CA). A therapeutic monoclonal antibody or a cocktail comprising a mixture of Individual monoclonal antibodies is prepared and used for the treatment of mice receiving subcutaneous or Sorthotopic injections of LAPC-9 prostate tumor xenografts.

Nl Cancer Xenorafts and Cell Lines SThe LAPC-9 xenograft, which expresses a wild-type androgen receptor and produces prostate-specific antigen 00 (PSA), is passaged in 6- to 8-week-old male ICR-severe combined immunodeficient (SCID) mice (Taconic Farms) by s.c.

trocar implant (Craft. et al., supra). The prostate carcinoma cell line PC3 (American Type Culture Collection) is

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maintained in RPMI supplemented with L-glutamine and 10% FBS.

Recombinant PC3 and 3T3- cell populations expressing a protein of the invention are generated by retroviral gene transfer as described in Hubert, et al., STEAP: a prostate-specific cell-surface antigen highly expressed in human prostate tumors. Proc Natj Acad Sd U S A, 1999. 96(25): p. 14523-8. Anti-protein of the invention staining is detected by using an FITC-conjugated goat anti-mouse antibody (Southem Biotechnology Associates) followed by analysis on a Coulter Epics-XL flow cytometer.

Xenoaraft Mouse Models.

Subcutaneous tumors are generated by injection of 1 x 10 6 LAPC-9, PC3, recombinant PC3-protein of the invention, 3T3 or recombinant 3T3-protein of the Invention cells mixed at a 1:1 dilution with Matrigel (Collaborative Research) in the right flank of male SCID mice. To test antibody efficacy on tumor formation, i.p. antibody injections are started on the same day as tumor-cell injections. As a control, mice are injected with either purified mouse IgG (ICN) or PBS; or a purified monoclonal antibody that recognizes an irrelevant antigen not expressed in human cells. In preliminary studies, no difference is found between mouse IgG or PBS on tumor growth. Tumor sizes are determined by vernier caliper measurements, and the tumor volume is calculated as length x width x height. Mice with s.c. tumors greater than 1.5 cm in diameter are sacrificed. PSA levels are determined by using a PSA ELISA kit (Anogen, Mississauga, Ontario). Circulating levels of, anti-109P1D4 protein mAbs are determined by a capture ELISA kit (Bethyl Laboratories, Montgomery, TX).

(See, Saffran, et al., PNAS 10:1073-1078 or www.pnas.org/cgi/doli10.1073/pnas.051624698) Orthotopic injections are performed under anesthesia by using ketamine/xylazine. For prostate orthotopic studies, an incision is made through the abdominal muscles to expose the bladder and seminal vesicles, which then are delivered through the incision to expose the dorsal prostate. LAPC-9 or PC3 cells (5 x 10 s mixed with Matrigel are injected into each dorsal lobe in a 10-pl volume. To monitor tumor growth, mice are bled on a weekly basis for determination of PSA levels.

The mice are segregated into groups for the appropriate treatments, with anti-protein of the invention or control mAbs being injected i.p.

Anti-109P1D4 Protein mAbs Inhibit Growth of Respective 109P1D4 Protein-Expressing Xenograft-Cancer Tumors The effect of anti-109P1 4 protein mAbs on tumor formation Is tested by using LAPC-9 and recombinant PC3protein of the invention orthotopic models. As compared with the s.c. tumor model, the orthotopic model, which requires injection of tumor cells directly in the mouse prostate or kidney, respectively, results In a local tumor growth, development of metastasis in distal sites, deterioration of mouse health, and subsequent death (Saffran, et al., PNAS supra; Fu, et al., 00 O Int J Cancer, 1992. 52(6): p. 987-90; Kubota, J Cell Biochem, 1994. 56(1): p. The features make the orthotopic Ci model more representative of human disease progression and allowed us to follow the therapeutic effect of mAbs on clinically relevant end points.

Accordingly, tumor cells are injected into the mouse prostate or kidney, and 2 days later, the mice are segregated into two groups and treated with either a) 200-500pg, of anti-109P1 4 protein Ab, or b) PBS three times per week for two to Sfive weeks.

A major advantage of the orthotopic prostate-cancer model is the ability to study the development of metastases.

Formation of metastasis in mice bearing established orthotopic tumors is studied by IHC analysis on lung sections using an

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antibody against a prostate-specific cell-surface protein STEAP expressed at high levels in LAPC-9 xenografts (Hubert, R.S., C et al., Proc Natl Acad Sd U S A, 1999. 96(25): p. 14523-8).

SMice bearing established orthotopic LAPC-9 or recombinant PC3-109P1D4 protein tumors are administered 00 1000pg injections of either anti-109P1D4 protein mAbs or PBS over a 4-week period. Mice In both groups are allowed to establish a high tumor burden (PSA levels greater than 300 ng/ml for IAPC-9), to ensure a high frequency of metastasis

C

formation in mouse lungs. Mice then are killed and their prostate and lungs are analyzed for the presence of tumor cells by IHC analysis.

These studies demonstrate a broad ant-tumor efficacy of anti-109P1 D4 protein antibodies on initiation and progression of prostate cancer in xenograft mouse models. Anti-109P1 D4 protein antibodies inhibit tumor formation of both androgen-dependent and androgen-independent tumors, retard the growth of already established tumors, and prolong the survival of treated mice. Moreover, anti-109P1D4 protein mAbs demonstrate a dramatic inhibitory effect on the spread of local prostate tumor to distal sites, even in the presence of a large tumor burden. Thus, anti-109P1D4 protein mAbs are efficacious on major clinically relevant end points (tumor growth), prolongation of survival, and health.

Example 39 Therapeutic and Dlaanostic use of Ant-1 09P D4 Antibodies In Humans, Anti-109P1 D4 monodonal antibodies are safely and effectively used for diagnostic, prophylactic, prognostic and/or therapeutic purposes in humans. Western blot and immunohistochemical analysis of cancer tissues and cancer xenografts with anti-109P1D4 mAb show strong extensive staining in carcinoma but significantly lower or undetectable levels in normal tissues. Detection of 109P1D4 in carcinoma and in metastatic disease demonstrates the usefulness of the mAb as a diagnostic and/or prognostic indicator. Anti-109P1D4 antibodies are therefore used in diagnostic applications such as immunohistochemistry of kidney biopsy specimens to detect cancer from suspect patients.

As determined by flow cytometry, anti-109P1D4 mAb specifically binds to carcinoma cells. Thus, anti-m09P1D4 antibodies are used in diagnostic whole body imaging applications, such as radiolmmunosdntigraphy and radioimmunotherapy, (see, Potamianos et al. Anticancer Res 20(2A):925-948 (2000)) for the detection of localized and metastatic cancers that exhibit expression of 109P1D4. Shedding or release of an extracellular domain of 109P1D4 into the extracellular milieu, such as that seen for alkaline phosphodiesterase B10 (Meerson, N. Hepatology 27:563-568 (1998)), allows diagnostic detection of 109P1D4 by anti-109P1D4 antibodies In serum and/or urine samples from suspect patients.

Anti-109P1D4 antibodies that specifically bind 109P1D4 are used in therapeutic applications for the treatment of cancers that express 109P1D4. Anti-109P1D4 antibodies are used as an unconjugated modality and as conjugated form in which the antibodies are attached to one of various therapeutic or imaging modalities well known in the art, such as a prodrugs, enzymes or radloisotopes. In preclinical studies, unconjugated and conjugated anti-109P1D4 antibodies are tested for efficacy of tumor prevention and growth inhibition In the SCID mouse cancer xenograft models, kidney cancer models AGS-K3 and AGS-K6, (see, the Example entitled "109P1D4 Monoclonal Antibody-mediated Inhibition of O Bladder and Lung Tumors In Vivo). Either conjugated and unconjugated anti-109P1 D4 antibodies are used as a therapeutic modality in human clinical trials either alone or in combination with other treatments as described in following Examples.

Example 40: Human Clinical Trials for the Treatment and Diagnosis of Human Carcinomas through use of Human Anti-109P1D4 Antibodles In vvo Antibodies are used in accordance with the present invention which recognize an epitope on 109P1D4, and are used in the treatment of certain tumors such as those listed in Table I. Based upon a number of factors, including 109P1D4 CK1 expression levels, tumors such as those listed in Table I are presently preferred indications. In connection with each of these S indications, three clinical approaches are successfully pursued.

Adjunctive therapy: In adjunctive therapy, patients are treated with anti-109P1D4 antibodies in 00 combination with a chemotherapeutic or antineoplastic agent and/or radiation therapy. Primary cancer targets, such as Sthose listed in Table I, are treated under standard protocols by the addition anti-109P1D4 antibodies to standard first and l 'second line therapy. Protocol designs address effectiveness as assessed by reduction In tumor mass as well as the ability to reduce usual doses of standard chemotherapy. These dosage reductions allow additional and/or prolonged therapy by reducing dose-related toxicity of the chemotherapeutic agent Anti-109P1D4 antibodies are utilized in several adjunctive clinical trials in combination with the chemotherapeutic or antineoplastic agents adriamydn (advanced prostrate carcinoma), cisplatin (advanced head and neck and lung cardnomas), taxol (breast cancer), and doxorubicin (predinical).

II.) Monotherapy: In connection with the use of the anti-109P1D4 antibodies in monotherapy of tumors, the antibodies are administered to patients without a chemotherapeutic or antineoplastic agent In one embodiment, monotherapy is conducted clinically in end stage cancer patients with extensive metastatic disease. Patients show some disease stabilization. Trials demonstrate an effect in refractory patients with cancerous tumors.

III.) Imaging Agent Through binding a radionudide iodine or yttrium (1131, Y9) to anti-109P1D4 antibodies, the radiolabeled antibodies are utilized as a diagnostic and/or imaging agent In such a role, the labeled antibodies localize to both solid tumors, as well as, metastatic lesions of cells expressing 109P1D4. In connection with the use of the anti-109P1D4 antibodies as imaging agents, the antibodies are used as an adjunct to surgical treatment of solid tumors, as both a pre-surgical screen as well as a post-operative follow-up to determine what tumor remains and/or returns.

In one embodiment, a 11 In)-109P1D4 antibody is used as an Imaging agent in a Phase I human clinical trial in patients having a carcinoma that expresses 109P1D4 (by analogy see, Divgi et al. J. Natl. Cancer Inst. 83:97-104 (1991)).

Patients are followed with standard anterior and posterior gamma camera. The results indicate that primary lesions and metastatic lesions are Identified.

Dose and Route of Administration As appreciated by those of ordinary skill in the art, dosing considerations can be determined through comparison with the analogous products that are in the clinic. Thus, anti-109P1 D4 antibodies can be administered with doses in the range of 5 to 400 mg/m 2 with the lower doses used, in connection with safety studies. The affinity of anti-109P1 D4 antibodies relative to the affinity of a known antibody for its target is one parameter used by those of skill in the art for determining analogous dose regimens. Further, anti-109P1D4 antibodies that are fully human antibodies, as compared to the chimeric antibody, have slower clearance; accordingly, dosing in patients with such fully human anti-109PID4 antibodies can be lower, perhaps in the range of 50 to 300 mg/m 2 and still remain efficacious. Dosing in mg/m 2 as opposed to the conventional measurement of dose In mg/kg, is a measurement based on surface area and is a convenient dosing measurement that is designed to include patients of all sizes from Infants to adults.

I

00 0 Three distinct delivery approaches are useful for delivery of anti-109P 04 antibodies. Conventional intravenous 0 delivery is one standard delivery technique for many tumors. However, in connection with tumors in the peritoneal cavity, such as tumors of the ovaries, biliary duct, other ducts, and the like, Intraperitoneal administration may prove favorable for obtaining high dose of antibody at the tumor and to also minimize antibody clearance. In a similar manner, certain solid tumors possess vasculature that is appropriate for regional perfusion. Regional perfusion allows for a high dose of antibody Sat the site of a tumor and minimizes short term clearance of the antibody.

Clinical Development Plan (CDP) Overview: The CDP follows and develops treatments of anti-109P1D4 antibodies in connection with adjunctive C therapy, monotherapy, and as an imaging agent Trials initially demonstrate safety and thereafter confirm efficacy in repeat N doses. Trails are open label comparing standard chemotherapy with standard therapy plus anti-109P104 antibodies. As will be appreciated, one criteria that can be utilized in connection with enrollment of patients is 109P1D4 expression levels in 00 their tumors as determined by biopsy.

As with any protein or antibody infusion-based therapeutic, safety concerns are related primarily to cytokine CK release syndrome, hypotension, fever, shaking, chills; (ii) the development of an immunogenic response to the material development of human antibodies by the patient to the antibody therapeutic, or HAHA response); and, (iii) toxicity to normal cells that express 109P1D4. Standard tests and follow-up are utilized to monitor each of these safety concerns.

Anti-109P1D4 antibodies are found to be safe upon human administration.

Example 41: Human Clinical Trial Adiunctive Therapy with Human Anti-109P1D4 Antibody and Chemotherapeutic Agent A phase I human clinical trial is initiated to assess the safety of six intravenous doses of a human anti-109P1 D4 antibody in connection with the treatment of a solid tumor, a cancer of a tissue listed in Table I. In the study, the safety of single doses of ant-109P1 D4 antibodies when utilized as an adjunctive therapy to an antineoplastic or chemotherapeutic agent as defined herein, such as, without limitation: cisplatin, topotecan, doxorubicin, adriamycin, taxol, or the like, is assessed. The trial design includes delivery of six single doses of an anti-109P1D4 antibody with dosage of antibody escalating from approximately about 25 mg/m 2 to about 275 mg/m 2 over the course of the treatment in accordance with the following schedule: Day Day 7 Day 14 Day21 Day28 mAb Dose 25 75 125 175 225 275 mg/m 2 mg/m 2 mg/m 2 mg/m 2 mg/m 2 mgm 2 Chemotherapy (standard dose) Patients are closely followed for one-week following each administration of antibody and chemotherapy. In particular, patients are assessed for the safety concerns mentioned above: cytokine release syndrome, hypotension, fever, shaking, chills; (ii) the development of an immunogenic response to the material development of human antibodies by the patient to the human antibody therapeutic, or HAHA response); and, (iii) toxicity to normal cells that express 109P1 D4. Standard tests and follow-up are utilized to monitor each of these safety concerns. Patients are also assessed for clinical outcome, and particularly reduction In tumor mass as evidenced by MRI or other Imaging.

00 0 The anti-109P1D4 antibodies are demonstrated to be safe and efficacious, Phase II trials confirm the efficacy and Srefine optimum dosing.

C) Example 42: Human Clinical Trial: Monotherapy with Human Anti-109P1D4 Antibody SAnti-109P1D4 antibodies are safe in connection with the above-discussed adjunctive trial, a Phase II human Sclinical trial confirms the efficacy and optimum dosing for monotherapy. Such trial is accomplished, and entails the same safety and outcome analyses, to the above-described adjunctive trial with the exception being that patients do not receive chemotherapy concurrently with the receipt of doses of anti-109P1D4 antibodies.

Example 43: Human Clinical Trial: Diagnostic Imagine with Anti-109PID4 Antibody Once again, as the adjunctive therapy discussed above is safe within the safety criteria discussed above, a human cl clinical trial is conducted concerning the use of anti-109P1D4 antibodies as a diagnostic imaging agent The protocol is 00 0 designed in a substantially similar manner to those described In the art, such as in Divgi et al. J. Natl. Cancerlnst. 83:97-104 0 (1991). The antibodies are found to be both safe and efficacious when used as a diagnostic modality.

Example 44: 109P1D4 Funcional Assays 1. Phosphorylation of 109PiD4 on tyrosine residues One hallmark of the cancer cell phenotype is the active signal transduction of surface bound receptor molecules, such as the EGF receptor, through tyrosine phosphorylation of their cytoplasmic domains and their subsequent interaction with cytosolic signaling molecules. To address the possibility that 109P1D4 is phosphorylated on its cytoplamsic tyrosine residues, 293T cells were transfected with the 109P1D4 gene in an expression plasmid such that the 109P1D4 gene was fused with a Myc/His tag, and were then stimulated with pervanadate (a 1:1 mixture of Na3VO4 and H202). After solubilization of the cells in Triton X-100, the 109P1D4 protein was immunopredpitated with anti-His polyclonal antibody (pAb), subjected to SDS-PAGE and Western blotted with anti-phosphotyrosine. Equivalent immunopredpitates were Western blotted with anti-His antibody. In Figure 22, 109P1D4 exhibits tyrosine phosphorylation only upon cell treatment with pervanadate and not without treatment This suggests that pervanadate, which inhibits intracellular protein tyrosine phosphatases (PTPs), allows the accumulation of phosphotyrosine (tyrosine kinase activity) on 109P1D4. Further, a large amount of the 109P1D4 protein is sequestered into the Insoluble fraction upon pervanadate activation, suggesting its association with cytoskeletal components. Similar effects of partial insolubility in Triton X-100 have been observed for cadherins, proteins that are related to protocadherins based on homology of their extracellular domains. Cadherins are known to interact with cytoskeletal proteins including actin, which are not readily soluble in the detergent conditions used in this study. Together, these data indicate that 109P1D4 is a surface receptor with the capacity to be phosphorylated on tyrosine and to bind to signaling molecules that possess SH2 or PTB binding domains, including but not limited to, phospholipase-Cyl, Grb2, Shc, Crk, PI-3-kinase p85 subunit, rasGAP, Src-family kinases and abl-family kinases. Such Interactions are important for downstream signaling through 109P1D4, leading to changes in adhesion, proliferation, migration or elaboration of secreted factors. In addition, 109P1D4 protein interacts with cytoskeletal components such as actin that facilitates its cell adhesion functions. These phenotypes are enhanced in 109P1D4 expressing tumor cells and contribute to their increased capacity to metastasize and grow in vivo.

Thus, when 109P1D4 plays a role in cell signaling and phosphorylation, it is used as a target for diagnostic, prognostic, preventative and/or therapeutic purposes.

Example 45: 109P1D4 RNA Interference (RNAI 00 0 RNA interference (RNAi) technology is implemented to a variety of cell assays relevant to oncology. RNA is a Spost-transcriptional gene silencing mechanism activated by double-stranded RNA (dsRNA). RNAi induces specific mRNA degradation leading to changes in protein expression and subsequently in gene function. In mammalian cells, these dsRNAs S caged short interfering RNA (siRNA) have the correct composition to activate the RNAi pathway targeting for degradation, specifically some mRNAs. See, Elbashir ef. Duplexes of 21-nucleotide RNAs Mediate RNA interference in 0Cultured Mammalian Cells, Nature 411(6836):494-8 (2001). Thus, RNAi technology is used successfully in mammalian cells to silence targeted genes.

Loss of cell proliferation control is a hallmark of cancerous cells; thus, assessing the role of 109P1D4 in cell r survival/proliferation assays Is relevant Accordingly, RNA was used to investigate the function of the 109P1D4 antigen. To rC generate siRNA for 109P1D4, algorithms were used that predict oligonudeotides that exhibit the critical molecular Sparameters (G:C content, melting temperature, etc.) and have the ability to significantly reduce the expression levels of the 00 109P1D4 protein when introduced into cells. Accordingly, three targeted sequences for the 109P1D4 siRNA are: AAAGAGGATACTGGTGAGATCT 3' (SEQ ID NO: 57)(oligo 109P1D4.a), 5' AAGAGCAATGGTGCTGGTAAA 3' (SEQ ID s NO: 58)(oligo 109P1D4.c), and 5' AACACCAGAAGGAGACAAGAT 3' (SEQ ID NO: 59)(oligo 109P1D4.d). In accordance with this Example, 109P1D4 siRNA compositions are used that comprise sIRNA (double stranded, short interfering RNA) that correspond to the nucleic add ORF sequence of the 109P1 D4 protein or subsequences thereof. Thus, slRNA subsequences are used in this manner are generally 5, 6, 7, 8, 9, 10, 11, 12,13,14, 15,16,17, 18, 19, 20, 21, 22, 23, 24, 26, 27, 28, 29, 30,31, 32, 33, 34, 35 or more than 35 contiguous RNA nucleotides in length. These siRNA sequences are complementary and non-complementary to at least a portion of the mRNA coding sequence. In a preferred embodiment, the subsequences are 19-25 nuceotides in length, most preferably 21-23 nucleotides in length. In preferred embodiments, these siRNA achieve knockdown of 109P1D4 antigen in cells expressing the protein and have functional effects as described below.

The selected siRNAs (109P1D4.a, 109P1D4.c, 109P1D4.d oligos) were tested in LNCaP cells in the 3 H-thymidine incorporation assay (measures cellular proliferation). Moreover, the oligonudeotides achieved knockdown of 109P1D4 antigen in cells expressing the protein and had functional effects as described below using the following protocols.

Mammalian siRNA transfections: The day before siRNA transfection, the different cell lines were plated in media (RPMI 1640 with 10% FBS wlo antibiotics) at 2x10 3 cells/well in 80 p (96 well plate format) for the proliferation assay.

In parallel with the 109P1 D4 specific siRNA oligo, the following sequences were included in every experiment as controls: a) Mock transfected cells with Upofectamine 2000 (Invitrogen, Carlsbad, CA) and annealing buffer (no siRNA); b) Luciferase-4 specific siRNA (targeted sequence: 5'-AAGGGACGAAGACGAACACUUCTT-3) (SEQ ID NO: 60); and, c) Eg5 specific slRNA (targeted sequence: 5'-AACTGAAGACCTGAAGACAATAA-3') (SEQ ID NO: 61). SiRNAs were used at 10nM and pg/ml Upofectamine 2000 final concentration.

The procedure was as follows: The siRNAs were first diluted In OPTIMEM (serum-free transfection media, Invitrogen) at 0.1 pM (10-fold concentrated) and Incubated 5-10 min RT. Upofectamine 2000 was diluted at 10 pg/ml fold concentrated) for the total number transfections and incubated 5-10 minutes at room temperature Appropriate amounts of diluted 10-fold concentrated Llpofectamine 2000 were mixed 1:1 with diluted 10-fold concentrated siRNA and Incubated at RT for 20-30" (5-fold concentrated transfection solution). 20 pis of the 5-fold concentrated transfection solutions were added to the respective samples and incubated at 37oC for 96 hours before analysis.

H-Thymidine incorporation assay: The proliferation assay is a H-thymidine incorporation method for determining the proliferation of viable cells by uptake and incorporation of label into DNA.

The procedure was as follows: Cells growing in log phase are trypsinized, washed, counted and plated in 96-well

OO

0 plates at 1000-4000 cells/well in 10% FBS. After 4-8 hrs, the media is replaced. The cells are incubated for 24-72 hrs, pulsed with 3 H-Thy at 1.5 pCi/ml for 14 hrs, harvested onto a filtermat and counted in scintillation cocktail on a Microbeta trilux or other counter.

L) In order to address the function of 109P1D4 in cells, 109P1D4 was silenced by transfecting the endogenously expressing 109P1D4 cell line (LNCaP) with the 109P1D4 specific siRNAs (109P1D4.a, 109P1D4.c, and 109P1D4.d) along with negative siRNA controls (Luc4, targeted sequence not represented in the human genome), a positive siRNA control (targeting Eg5) and no siRNA oligo (LF2K) (Figure 23). The results indicated that when these cells are treated with siRNA specifically targeting the 109P1D4 mRNA, the resulting '109P1D4 deficient cells' showed diminished cell proliferation as C measured by this assay see oligo 109P1D4.a treated cells).

SThese data indicate that 109P1 D4 plays an important role in the proliferation of cancer cells and that the lack of 109P1D4 dearly decreases the survival potential of these cells. It is to be noted that 109P1D4 is constitutively expressed in 00 many tumor cell lines. 109P1D4 serves a role in malignancy; Its expression is a primary indicator of disease, where such Sdisease is often characterized by high rates of uncontrolled cell proliferation and diminished apoptosis. Correlating cellular C phenotype with gene knockdown following RNA treatments is important, and allows one to draw valid conclusions and rule out toxicity or other non-specific effects of these reagents. To this end, assays to measure the levels of expression of both protein and mRNA for the target after RNA treatments are important, including Western blotting, FACS staining with antibody, Immunopredpitation, Northern blotting or RT-PCR (Taqman or standard methods). Any phenotypic effect of the siRNAs in these assays should be correlated with the protein and/or mRNA knockdown levels in the same cell lines.

109P1D4 protein is reduced after treatment with siRNA ollgos described above 109P1D4.a, etc.) A method to analyze 109P1 D4 related cell proliferation is the measurement of DNA synthesis as a marker for proliferation. Labeled DNA precursors 3 H-Thymidine) are used and their incorporation to DNA is quantified.

Incorporation of the labeled precursor into DNA is directly proportional to the amount of cell division occurring in the culture.

Another method used to measure cell proliferation is performing clonogenic assays. In these assays, a defined number of cells are plated onto the appropriate matrix and the number of colonies formed after a period of growth following siRNA treatment is counted.

In 109P1D4 cancer target validation, complementing the cell survival/proliferation analysis with apoptosis and cell cycle profiling studies are considered. The biochemical hallmark of the apoptotic process is genomic DNA fragmentation, an irreversible event that commits the cell to die. A method to observe fragmented DNA in cells is the immunological detection of histone-complexed DNA fragments by an immunoassay cell death detection ELISA) which measures the enrichment of histone-complexed DNA fragments (mono- and oligo-nudeosomes) in the cytoplasm of apoptotic cells. This assay does not require pre-labeling of the cells and can detect DNA degradation in cells that do not proliferate in vitro freshly isolated tumor cells).

The most important effector molecules for triggering apoptotic cell death are caspases. Caspases are proteases that when activated cleave numerous substrates at the carboxy-terminal site of an aspartate residue mediating very early stages of apoptosis upon activation. All caspases are synthesized as pro-enzymes and activation involves deavage at aspartate residues. In particular, caspase 3 seems to play a central role in the initiation of cellular events of apoptosis.

Assays for determination of caspase 3 activation detect early events of apoptosis. Following RNAI treatments, Western blot detection of active caspase 3 presence or proteolytic cleavage of products PARP) found In apoptotic cells further support an active induction of apoptosis. Because the cellular mechanisms that result in apoptosis are complex, each has its advantages and limitations. Consideration of other criteria/endpoints such as cellular morphology, chromatin condensation, membrane blebbing, apoptotic bodies help to further support cell death as apoptotic. Since not all the gene targets that 00 00 regulate cell growth are anti-apoptotic, the DNA content of permeabilized cells is measured to obtain the profile of DNA content or cell cycle profile. Nuclei of apoptotic cells contain less DNA due to the leaking out to the cytoplasm (sub-G1 population). In addition, the use of DNA stains propidium iodide) also differentiate between the different phases of the cell cycle in the cell population due to the presence of different quantities of DNA In GO/G1, S and G2/M. In these studies the subpopulations can be quantified.

For the 109P1 D4 gene, RNAi studies facilitate the understanding of the contibution of the gene product in cancer pathways. Such active RNAi molecules have use in Identifying assays to screen for mAbs that are active anti-tumor therapeutics. Further, sIRNA are administered as therapeutics to cancer patients for reducing the malignant growth of Cseveral cancer types, Including those listed In Table I. When 109P1D4 plays a role in cell survival, cell proliferation, tumorigenesis, or apoptosis, it is used as a target for diagnostic, prognostic, preventative and/or therapeutic purposes.

00 Throughout this application, various website data content, publications, patent applications and patents are referenced. (Websites are referenced by their Uniform Resource Locator, or URL, addresses on the World Wide Web.) cThe present Invention Is not to be limited in scope by the embodiments disclosed herein, which are intended as single illustrations of individual aspects of the Invention, and any that are functionally equivalent are within the scope of the Invention. Various modifications to the models and methods of the invention, in addition to those described herein, will become apparent to those skilled In the art from the foregoing description and teachings, and are similarly intended to fall within the scope of the Invention. Such modifications or other embodiments can be practiced without departing from the true scope and spirit of the invention.

00 00

TABLES:

TABLE I: Tissues that Express 1O9PD4 when malignant: Prostate Bladder Ildney Colon Lymphoma Lung Pancreas Ovary Breast Uterus Stomach Rectum Cervix Lymph Node Bone TABLE i: Amino Acid Abbreviations SINGLE LETTER THREE LETTER FULL NAME F Phe phenylalanine L Leu leucne S Ser serine Y Tyr tyrosine C CYS cysteine W Trp tytophan P Pro proline H His hisfidine Q Gin glutamine R Arg arginine lie isoleucine M Met methionine T Thr threonine N Asn asparagine K Lys lysine V Val valine A Ala alanine D Asp aspartic acid E Gu glutamic acid G I Gly glycIne 00 TABLE III: Amino Acid Substitution Matrix Adapted from the GOG Software 9.0 BLOSUM62 amidno adid substitution matrix (block substitution matrix). The a) higher the value, the more likely a substitution is found in related, natural proteins. (See world wide web URL ikp.unibe.ch/manual/blosum62.html) A C D E F G H I1K L M N P Q R S T V W Y.

4 0 -2 -1-2 0 -2-1 -1-1 -1 -2 -1-1-1 1 0 0 -3 -2 A 9 -3 -4 -2 -3 -3 -1 -3 -1 -1 -3 -3 -3 -3 -1 -1 -1 -2 -2 C 6 2 -3-1 -1-3 -1 -4-3 1 -1 0 -2 0 -1-3 -4-3 D -3 -2 0 -3 1 -3 -2 0 -1 2 0 0 -1-2 -3-2 E 6 -3 -1 0 -3 0 0 -3-4 -3-3 -2 -2-1 1 3 F 6~ 6-2 -4-2 -4-3 0 -2 -2-2 0 -2-3 -2-3 G 8 -3 -1-3 -2 1 -2 0 0 -1 -2-3 -2 2 H 4~ 4-3 2 1-3 -3-3 -3 -2-1 3 -3-1 1 005-2- 0- 1 2-3-2 4 2 -3 -3-2 -2 -2-1 1 -2 -1 L 5-2 -2 0 -1-1 -1 1 -1 -1M 6 -2 0 0 1 0 -3 -4 -2 N 7 -1 -2 -1 -1 -2 -4 -3 P 1 0 -1-2 -2-1 Q -1 -1 -3 -3 -2 R 4 1 -2 -3 -2 S 0 -2 -2 T 4 -3 -1 V 11 2 W 7 Y TABLE IV: HLA Class 11 Motfs/Supermofifs TABLE IV HLA Class I Supermotgsllotifs SUPERMOTIF POSITION POSITION POSITION 2 (Primary Anchor) 3 (Primary Anchor) C Terminus (Primary Anchor) Al TILVMS FWY A2 LWIATQ VMATL A3 VSMATU_ RK A24 YFWIVLMT FIYXMI B7 P VILFMWYA 827 RHK FYLWMIVA 844 ED FWYUMVA 858 ATS FWYUVMA 862 QLVMP FWYMIVLA

MOTIFS

Al TSM Y Al IDEAS Y A2.1 LMVQIAT VIMAT A3 LMVISATFCGD KYRHFA All VTMLISAGNCDF KRYH A24 YFWM_ FLIW A'3101 MTAUS RK A'3301 MVALFIST RK

A

t 6801 AVTMSL_ RK 8*0702 P LMFWYAIV 8*3501 P LMFWY/VA B51 P LIVEWYAM B'5301 P IMFWYALV 5*5401 P ATIVLMFWY Bolded residues are preferred, Italicized residues are less preferred: A peptide is considered motif-bearing if it has primary anchors at each primary anchor position for a motif or supermotif as specified In the above table.

TABLE IV HLA Class II Superoxtif 1 6 9 W, F, Y. L A V, 1. L, P, C, S, T AV, 1, L, C, S, T, M, Y 00 TABLE IV HLA Class II Motifs MOTIFS 1l*anchor 1 2 3 4 5 1 anchor 6 7 8 9 DR4 preferred FIfYUVW M T I VSTCPALJM MH MH deleterious W R WDE DRI preferred MFLJVWY PAMQ VMATSPLIC M AVM deleterious C CH FD CWD GDE D DR7 preferred MFIWY M W A IVMSACTPL M IV deleterious C G GRD N G MOTIFS 1l anchor i 2 3 1*anchor4 5 1l 0 anchor 6 Motif a preferred UVMiFY D Motif b preferred LIVMFAY DNQEST KRH DR Supermotif MFUVWY VMSTACPU Italicized residues Indicate less preferred or tolerated' residues TABLE IV HLA Class I Supermotfs POSITION: I 3 4 5 6 7 8 C-terminus

SUPER-

MOTIFS

Al V Anchor i2A9r TILVM4S FWY A2 1 Anchor 1 Anchor LNMATQ UVMAT A3 Preferred I'Anchor YFW YFW YFW P IQ Anchor VSMATLI RK deleterious DE DE (415) A24 I*Ancho, 1*Anchor YFMVLM T FIYWLM B7 Preferred FWY 1 hlr FWY FWY I Anctor UVM P VLFMWYA deleterious DE DE G ON DE (315) (415) B27 10 An1And~ RHK FYLWMIVA 844 1*Anchor I ED FWYIUMVA 858 I Pch 1 0 Anchor ATS FWYLIVMA 862 10 r 1Anchor QUVMP FWYMIVLA Italicized residues indicate less preferred or tolerated" residues 00 TABLE IV HLA Class I Motifs POSITON I 3 4 5 6 7 8 9 00 term dnus or C-termrinus Al preferred GFYW Il 0 Andio OEA YFW P DEON YFW I An 9-nier STM y deleterious DE RHKUVMP A G A Al preferred GRHK ASTCUVM l 0 Andior GSTC ASTO UVM DE l 0 Anchor 9-mner DEAS

Y

deleterious A RHKDEPYFW DE PQN RHK PG GP Al preferred YFW I -Anhor DEAQN A YFWQN PASTC GDE P 1 oAnchor STh4 y mer deleterious GP RHKGUVM DE RHK QNA RHKYFW RHK A Al preerd YFW STCUVM 11 Aa~a A YFW PG G YFW l 0 Anchc DEAS

Y

mer deleterious RHK RHKDEPYFW P G PRHK QN A21 preferred YFW I Aco YFW STC YFW A P I 0,kr 9-4mr LMJVQAT

VUIMAT

deleterious DEP DERKH RKH DERKH POSITION:l1 2 3 4 5 6 7 8 9 c- Terminus A21 preferred AYFW I Anchor LVIM G G FYWL 1*Anch2 LMIVQP4T vim VUIMAT mer deleterious DEP DE RKH-A P RKH DERKHRKH A3 preferred RHK l~I*Aor YFW PRHKYF A YFW P I *Anchor LMVISATFCGD W KYRHFA deleterious DEP DE All preferred A I!nco YFW YFW A YFW YFW P P*Azchor VTLMISAGNCD

KRW-I

F

deleterious DEP A G A24 preferred YPVVRHK I An c- STC YFW YFW I2!l2 9-mer YFWM

FLIW

deleterious DEG DE G QNP DERI-KG AQN A24 Preferredmc P, YFWP P YFWM

FLAW

mer Deleterious GDE QN RHK DE A QN DEA A3101 Referred RH-K J*nc YFW P YFW YFW PP I!AncQr MVTAIJS

RK

Deleterious DEP DE ADE DE DE DE A3301 Preferred 1rAnho YFW AYFW onh MVALFIST

RK

Deleterious GP DE A6801 Preferred YFWSTC l 0 Anchor YFWLIV YFW P l 0 Pnchor AVTMSLI M RK deleterious GP DEG RHK A B0702Preferred RHKFWY l*Arichor RHK RHK RHK RHK PA l 0 Pnchor P

LMFWYAJ

V

deleterious DEQNP DEP DE DE GDE QN DE 00 00 POSITION 1 3 4 5 6 7 8 9 Cor C-termninus All prefbrred GEYW j I IDEA YFW P DE(N YFW 1 Anho 9-mer STM

Y

deleterious DE Rin1VMP A G A Al preferred GRHK ASTCUVM I *Anchor GSTC ASTC LIVM DE I 0 Anchor 9-mr IDEAS

Y

deleterious A RHKDEPYFW DE PQN RHK PG GP B350lPrefffred FWYLIVM I &nch FWY FWY lAnchor P

LMFWYIV

A

deleterious AGP G G 651 PreWeWe LMIFWY IJonh FWY STC FWY G FWY' PAnchor P

UVFWYA

M

deleteios AGPIDER DE G DEQN GDE

HKSTC

85301 preferre UVMFWY 1 *Anchor FWY STC FWY LIVMFWY'FWY I 0 Anchor P

IMFWYAL

V

deleterious AGPQN G RHKQN DE B5401 prefere FWY loAnw FWY'LIVMV LIVM ALIVM F'WYA l Andior P P A1WLMF deLteiou GlPQNDE GDESTC RHKDE DE QNDGE DE 00 00 TABLE IV ummary of HLA-supertyps rail phienotypic frequencies of IILA-supertypes In different ethnic populations ____Specificity Phenotyic fireuen___- pryeosition 2 erinu caslanINA Bla apanese ~in I s an erage 7 3.2 5.1 1 3.0 9.3 _____ILMvVST 7.5 2.1 5.8 2. 3.1 .2 1 ____ILMVTr LMVTr 5.8 19.0 12.4 5.9 3.0 2.2 4 F (WIL I 3.9 .9 .6 .1 8.3 0.0 IMVA 3.0 1.2 42.9 9.1 9.0 I LVMS 47.1 16.1 21.8 14.7 6.3 5 .2 27 HK L 1 28.4 .1 13.3 3.9 5.3 3.4 62 LIVMP) MI 12. .8 36.5 5.4 11.1 8.1 58 TS LIV 10.0 5.1 1.6 .0 .9 0O.3 TABLE IV HLA-supertypes Phenotypic frequency ucasian .A Blacks Japanese Phinese ispanic: vea .0 .1 87.5 08.4 .3 .2 A2, A3 and B7 r9.5 8.1 100.0 9.5 .4 .3 A2, A3, B7, A24, .9 .6 100.0 9.8 9.9 .8 and Al A2, A3, B7, P24, B44, Al, 827, 862, and B 58 Motifs indicate the residues defining supertype spectficites. The motifs incorporate residues determined on the basis of published data to be recognized by multiple alleles within the supertype. Residues within brackets are additional residues also predicted to be tolerated by multiple alleles within the supertype.

Table V. Frequently Occurring Motifs Name dentit Dscription Potential Function qucielc acid-binding protein functions as Yanscription facor, nuclear location Zin finger, C21-2 type obable Cytochrome b(N- mmbrane bound oxidase, generate L68% erminal)1b6/etB sperodde morains; are one hundred amino acids ong and include a conserved IQ 19% Immunoglobulln domain ntradomaln disulfide bond.

:andem repeats of about 40 residues, Dach containing a Trp -Asp motif.

:unction In signal transduction and 18% WDdomain, G-beta repeal rotein interaction may function in targeting signaling 23% PDZ domain molecules to sub-membranous sites -R 8% Leudne Rich Repeat shart sequence motifs Involved In Drotein-protein interactions nserved catalytic core common to lI serine/threonine and tyrosine rotein kinases containing an ATP knasa 3% Protein kInase domain inding site and a catalytic site 00 00 pleckstrin homology involved in ntracelluiar signaling or as constituents P16% PH domain )f the cytoskeleton 30-40 amino-acid long found in the axlraceliuiar domain of membrane- 34% 1GF-like domain bond proteins or in secreted proteins Reverse transcriptase (RNA-dependent DNA 49% polymerase) ,ytoplasmic protein, associates integral An 5% jk repeat -embrarie proteins to the cytoskeleton NAH- nmbrane associated. Involved in Ubiquinonelplastoquinone xton translocation across the xiddoredgql 32% (complex various chains Timbrane xdcum-binding domain, consists of a12 esidue loop flanked on both sides by a 24% 12 residue alpha-helical domain Retroviral aspartyl ksatlor acid proteases, centered on 79% )rotease 3catalytic aspartyl residue )xtraceiiuiar structural proteins Involved n formation of connective tissue. The Colagen triple helix repeat ieuence consists of the G-X-Y and the Collagen 42% (20 copies) olypeptide chains forms a triple helix.

-ocated in the extracellular ligand- Anding region of receptors and is about amino acid residues long with two )airs of cysteines Involved in disulfide 20% ribronectin type IIl domain 3nds seen hydrophobic transmembrane ions, with the N-terminus located 7transmembrane receptor xtraceliularly while the C-terminus is 7tm_1 19% (rhodopsin family) opasmic. Signal through G proteins Table VI: Post-translational modifications of 109P1D4 Ooiyco~aion sites 231 S 238 S 240 T 266 T 346 T 467 T 551 T 552 S 555 T 595 T 652 S 654 S 660 T 790 T 795 T 798 T 804 S 808 S 923 T 927 T 954 T 979 S 982 S 983 8 00 985 S 986 S C~1 990 S 999 T IODO T 1006 S 1017 S 1020 T Serine poshorvation sites DLNLSLIPN (SEQ ID NO: 62) 147 VINISIPEN (SEQ ID NO: 63) 152 IPENSAINS (SEQ ID NO: 64) 238 ILQVSVTDT (SEQ ID NO: 257 EIEVSIPEN (SEQ ID NO: 66) 428 LDYESTKEY (SEQ ID NO: 67) 00 480 PENNSPGIQ (SEQ ID NO: 68) 489 LTKVSAWDA (SEQ ID NO: 69) 495 MDADSGPNA (SEQ ID NO: 559 TVFVSIIDQ (SEQ ID NO: 71) 567 QNDNSPVFT (SEQ ID NO: 72) 608 AVTLSILDE (SEQ ID NO: 73) 630 RPUISFDRE (SEQ ID NO: 74) 638 EKQESYTFY (SEQ ID NO: 652 GGRVSRSSS (SEQ ID NO: 76) 654 RVSRSSSAK (SEQ ID NO: 77) 655 VSRSSSAKV (SEQ ID NO: 78) 656 SRSSSAKVT (SEQ ID NO: 79) 714 EVRYSIVGG (SEQ ID NO: 789 LVRKSTEAP (SEQIDNO: 81) 805 ADVSSPTSD (SEQ ID NO: 82) 808 SSPTSDYVK (SEQ ID NO: 83) 852 NKQNSEWAT (SEQ ID NO: 84) 877 KKKHSPKNL (SEQ ID NO: 898 DDVDSDGNR (SEQ ID NO: 86) 932 FKPDSPDLA (SEQ ID NO: 87) 941 RHYKSASPQ (SEQ ID NO: 88) 943 YKSASPQPA (SEQ ID NO: 89) 982 ISKCSSSSS (SEQ ID NO: 983 SKCSSSSSD (SEQ ID NO: 91) 984 KCSSSSSDP (SEQ ID NO: 92) 985 CSSSSSDPY (SEQ ID NO: 93) 990 SDPYSVSDC (SEQ ID NO 94) 1006 EVPVSVHTR (SEQ ID NO: Threonine Dhosohordeadon sites 29 EKNYTIREE (SEQ ID NO: 96) 81 IEEDTGEIF (SEQ ID NO: 97) 192 DVMETPEGD (SEQ ID NO: 98) 252 VFKETEIEV (SEQ ID NO: 99) 310 TGLITIKEP (SEQ ID NO: 100) 320 DREETPNHK (SEQ ID NO: 101) 551 VPPLTSNV (SEQ ID NO: 102) 790 VRKSTEAPV (SEQ ID NO: 103) 856 SEWATPNPE (SEQ ID NO: 104) 924 NWVTrTPTTF (SEQ ID NO: 105) 927 TTPTTFKPD (SEQ ID NO: 106) 999 GYPVTFEV (SEQ ID NO: 107) 1000YPVTTFEVP (SEQ ID NO: 108) Twfosine phospholallon sites 67 FKLVYKTGD (SEQ ID NO 109) 00 00 158 INSKYTLPA (SEQ ID NO: 110) 215 EKDTYVMKV (SEQ ID NO: I111) 359 IDIRYIVNP (SEQ ID NO: 112) 423 ETMAYLDYE (SEQ ID NO: 113) 426 AYLDYESTK (SEQ ID NO: 114) 432 STKEYAIKL (SEQ ID NO: 115) 536 KEOKYLFTI (SEQ ID NO: 116) 599 TDPDYGDNS (SEQ ID NO, 117) 642 SYTFYVKAE (SEQ ID NO: 118) 682 SNCSYELVL (SEQ ID NO: 119) 713 AEVRYSIVG (SEQ ID NO: 120) 810 PTSOYVKIL (SEQ ID NO: 121) 919 TMGKYNWVT (SEQ ID NO: 122) 989 SSDPYSVSD (SEQ ID NO: 123) 996 SDCGYPV1T (SEQ ID NO: 124) Table VII: Search Peodes 109P1D4 v.1 9-mers, lO-mers and 15-mrs (SEQ ID NO: 125) MDLLSGTYIF AVLLACVVFH SGAQEKNYTI MQFKLVYKTG DVPLIRIEED TGEIE'rTGAR RLVKIRFLIE DINDNAPLFP ATVINISIPE SQNIFGLDVI ETPEGDKMPQ LIVQRELDRE DTNDNHPVFK ETEIEVSIPE NAPVGTSVTQ HLNATTGLIT IKEPLDREET PNHKLLVLAS VNPVNDTVVL SENIPLNTKI ALITVTDKDA AAYLDYESTK EYAIKLLAAD AGKPPLNQSA PGIQLTKVSA MDADSGPNAI( INYLLGPDAP

REEMPENVLI

IDREKLCAGI

NSAINSKYTL

EKDTYVMKVK

LHATDADIGE

DGGU4PARAM

DHNGRVTCFT

MLFIKVKDEN

GDLLKDLNLS LIPNKSLTTA PRDEHCEfYEV EVAILPDEIF PAAVDPDVGI NGVQNYELII< VEDGGFPQRS STAILQVSVT NAKIHFSFSN LVSN4IAPRRLF VLVNVTDVND NVPSIDIRYI DHEIPFPLRP VFSNQFLLET DNAPV M SF VTVSIPENNS LAKDKGVPPL TSNVTVFVSI DNSAVTLSIL DENDEIDS INVVDVNDNK PVFIVPPSNC RDLFAIDQET GNITLMEKCD LINELVRXST EAPVTPNTEI HLKAAQKcNKQ NSEWATPNPE NRVTLDLPID LEEOTMGKYM HIIQELPLDN TFVACDSIS(

F

IDQNDNSPVF

QTGVIRPNIS

SYELVLPSTN

VTDLGLHRVL

ADVSSPTSDY

NRQMIMMKKK

WVTTPTTFKP

CSSSSS5DPYS PEFSLDCRTG MLTVVKKLDR EKEDiCYLFTI THNEYNFYVP ENLPRHGTVG LITVTDPDYG FDREKQESYT FYVKPIEDGGR VSRSSSAKVT PGTVVFOVIA VDNDTGMNAE VRYSIVGGNT VKANDLGQPD SLFSVVIVNL FVNESVTNAT VKILVAAVAG TITVVVVIFI TAVVRCRQAP KKKKKHSPRN LLLNFVTIEE TKADDVDSDG DSPDLAD.HYK SASPQPAFQI QPETPLNSKH VSDCGYPVTT FEVPVSVHTR PVGIQVSNTT 540 600 660 720 780 840 900 960 1020 1021 109P1 D4 v.2 (both ends diff frm v. 1) N' terminal 9-mers aa -30 to 8 MRTERQWVIJIQIFQVLCGLIQQTVTSVPGMDLLSGTY (SEQ ID NO: 126) aa -30 to 9 MRTERQWVLIQIFQVLCGLIQ-QTVTSVPGMDJLSGTYI (SEQ ID NO: 127) aa-30Oto 14 MRTERQWVLIQIFQVLCGLIQQTVTSVPGMDLLSGTYIFAVLL (SEQ ID NO: 128) 109P1D4 v.2 C' Termninal 9 mes: aa 1004 to 1025 PVSVHTRPTDSRTSTIEICSEI (SEQ ID NO: 129) mers: aa1003 to 1025 VPVSVHTRPTDSRTSTIEICSEI (SEQ ID NO: 130) mers: aa 997 to 1025 VTTFEVPVSVHiTRPTDSRTSTIEICSEI (SEQ ID NO: 131) 109P1 D4 v.3 9 mers: aa 1004 to 1347 (SEQ ID NO: 132)

PVSVHTRPPMKEVVSCTPMESTTMEIWIHPQPQJCSEGKVAGRSQPJRVTFHLPEGSQESSSDG

GLGDHDAGSLTSTSHGLPLGYPQEEYFDRATPSNRTEGIGNSDPESTFI

PGLKKAAEITVQPTVE

EASDNCTQECLIYGHS DACWMPASLDHSSSSQAQASALCHSPPLSQASTQHHSPRVTQTIALCHS

PPTTACSPIVAHSPVAAHSPAAACSPAAASSPP

VIALHRSQAQSSVSLQQGWVQGADGLCSVDQGVQGSATSQFYTMSERLHPSDDSIKVI

PLTTFTP

RQQARPSRGDSPMEEHPL

mers: aa 1003 to 1347 (SEQ ID NQ 133) VPVSVHTRPPMKEVVRSCTPMKESTTNEIWIHPQPQRKSEGECVAGKSQRRVTFHILPEGSQESSS 0

GGLGDHDAGSLTSTSHGLPLGYPQEEYFDRATPSNRTEGDGNSDPESTFIPGLKKAAEITVQPTV

EEASDNCTQECLIYGHSDACWMPASLDHSSSSQAQASALCHSPPIJSQASTQHHSPRVTQTIALCH

SPPVTQTIALCHSPPPIQVSALHHSPPLVQATALHHSPPSAQASALCYSPPLiAQAAAISHSSPLP QVIALHRSQAQSSVSLQQGWVQGADGLCSVDQGVQGSAESQFYTMSERLHPSDDS IKVI PLTTFT PRQQARPSRGDS PMEEHPL mets: aa 998 to 1347 (SEQ ID N0r 134) VTTFEVPVSV HTRPPNKEVV RSCTPMKEST FEGSQESSSD GGLGDHDAGS LTSTSHGLPL PGLKKAAEIT VQPTVEEASD NCTQECLIYG SQASTQHHSP RVTQTIALCH SPPVTQTIAL AQASALCYSP PLAQAAAISH SSPLPQVIAJ GSATSQFYTM SERILHPSDDS IKVIPLTTFT

TMEIWIHPQP

GYPQEEYFDR

HSDACWMPAS

CHSPPPIQVS

HRSQAQSSVS

PRQQARPSRG

QRKSEGKVAG

ATPSNRTEGD

LDHSS SSQAQ

ALHHSPPLVQ

LQQGWVQGAD

DSPMEEHPL

KSQRRVT FHIL

GNSDPESTFI

ASALCHSPPL

ATALHHS PPS

GLCSVDQGVQ

109P1D4 v.4 (deleting 10 aa, 1039-1048, from v.1) 9-mets aa 1031-1056 (deleting 10 aa, 1039-1048, from v.1) IWIHPQPQSQRRVTFH (SEQ ID NO: 135) aa 1030- 1057 (deleting 10 aa, 1039-1048, from v.1) EIWIHPQPQSQRRVTFHL (SEQ ID NO: 136) aa 1025- 1062 (deleting 10 aa, 1039-1048, from v.1) ESTTMEIWIHPQPQSQRRVTFHLPEGSQ (SEQ ID NO: 137) 109P1 D4 v.5 (deleting 37 aa, 1012-1048, from v.1) 9-mets aa 1004-1056 (deleting 37 aa, 1012-1048, from v.1) PVSVHTRPSQRRVTFH (SEQ ID NO: 138) aa 1003-1057 (deleting 37 aa, 1012-1048, from v.1) VPVSVHTRPSQRRVTFHL (SEQ ID NO: 139) aa 998-1062 (deleing 37 aa, 1012-1048, from v.) VTTFEVPVSVHTRPSQRRVTFHLPEGSQ (SEQ ID NO: 140) 109P1 D4 v.6 (both ends diff from v. 1) N' terminal 9-mets: aa -23 to 10 (excluding I and 2) MTVGFNSDISSVVRVNTTNCHKCLLSGTYIF (SEQ ID NO: 141) 1 0-mets: aa -23 to 11 (excluding I and 2) MTVGFNSDISSVVRVNTTNCHKCLLSGTYIFA (SEQ aa -23 to 17 (excluding 1 and 2) MTVGFNS DI SSVVRVNTTNCHKCILSGTYI FAVLLVC ID NO: 142) (SEQ ID NO: 143) 109PI1D4 v.6 C' terminal 9-mets: aa 1004-1016 PVSVHTRPTDSRT (SEQ ID NO: 1 0-mars: aa 1003-1016 VPVSVHTRPTDSRT (SEQ ID NO: 144) 145) VTTFEVPVSVHTRPTDSRT (SEQ ID NO: 146) 1 09131 D4 v.7 (N-terminal 21 aa duff from tose in v.6) 00 N' terminal 9-mers aa -21 to 10 (excluding 1 and 2) rFRVGFLIISSSSSLSPLLLVSVVRVNTT (SEQ ID NO: 147) aa -21 to 11 (excluding 1 and 2) MFRVGFLIISSSSSLSPLLLVSVVRVNTTN (SEQ ID NO: 148) aa -21 to 16 (excluding I and 2) MFRVGFLIISSSSSLSPLLLVSVVRVNTTNCHKCL (SEQ ID NO: 149) 109P1D4 v.8 9-mers aa 1099-1126 (excuding1117 and i 11) (71 TFIPGLKKEITVQPTV (SEQ ID NO: 150) l-mers aa 1098-1127 (excluding 1117land 1118) STFIPGLKKEITVQPTVE (SEQ ID NO: 151) 00 15-mers aa 1093-1131 (excluding 1117 and 1118) NSDPESTFIPGLKKEITVQPTVEEASDN (SEQ ID NO: 152) 109P10D4 v. 1, v.2 and v.3 SNP variants 9-fners TYIFAVLLVCVVFHSGA (SEQ ID NO; 153) lO-mers GTYIFAVLLVCVVFHSGAQ (SEQ ID NO: 154) MDLLSGTYIFAVLLVCVVFHSGAQEKNYT (SEQ ID NO: 155) 109P1D4 v.1, v.2 and v.3 SNP variants M341 9-mers KNYTIREEIPENVLIGD (SEQ ID NO: 156) 1 0-mets EKNYTIREEIPENVLIGDL (SEQ ID NO: 157) I HSGAQEKNYTIREEIPENVLIGDLLKDLN (SEQ ID NO: 158) 109P1D4 v.1, v.2 and v.3 SNP variants M341 and D42N 94ms KNYTIREEIPENVLIGN (SEQ ID NO: 159) -mers EKNYTIREEIPENVLIGNL (SEQ ID NO: 160) HSGAQEKNYTIREEIPEWVLIGNLLKDLN (SEQ ID NO: 161) 109P1D4 v0, Y.2 and v.3 SNP variants D42N 9-mrs MPENVLIGNLLKDLNLS (SEQ ID NO: 162) I 0-mts EMPENVLIGNLLKDLNLSL (SEQ ID NO: 163) YTIREEMPENVLICGNLLKDLNLSLIPNKS (SEQ ID NO: 164) 10991D4 v.1, v.2 and v.3 SNP variants D42N and M341 94mets IPENVLIGNLLKDLNLS (SEQ ID NO: 165) 104-m EIPENVLIGNLLKDLNLSL (SEQ ID NO: 166) 00 YTIREEIPENVLIGNLLKDLNLSLIPNKS (SEQ ID NO: 167) I109PI1D4 v.1, v.2 and v.3 SNP variants 9-mers IPNKSLTTTMQFKLVYK (SEQ ID NO: 168) 1lO-mers LIPNKSLTTTMQFKLVYKT (SEQ ID NO: 169) DLNLSLIPNKSLTTTMQFKLVYKTGDVPLI (SEQ ID NO: 170) ri 109P1 D4 v.1, v.2 and v.3 SNP variants 1154V C1 9-mers ISIPENSAVNSKYTLPA (SEQ ID NO: 171) 001 0-mers 00 NISIPENSAVNSlKYTLPAA (SEQ ID NO: 172) PATVINISIPENSAVNSKYTLPAAVDPDV (SEQ ID NO: 173) 109P1D4 v.1, v.2 and v.3 SNP variants V2921 9-mars IHFSFSNLISNIARRLF (SEQ ID NO: 174) 1O-mers KIHFSFSNLISNIARRLFH (SEQ ID NO: 175) IGENAKIHFSFSNLISNIAP.PLFHLNATT (SEQ ID NO: 176) 109P1D4 v.1, v.2 and v.3 SNP variants T420N 9-mers FSNQFLLENAAYLDYES (SEQ ID NO: 177) lO-mers VFSNQFLLENAAYLDYEST (SEQ ID NO: 178) FRLRPVFSNQFLLENAAYL DYES TKE YAI (SEQ ID NO: 179) 109P1D4 v.1, v.2 and v.3 SNP variants T486M 9-mers NNSPGIQLMKVSAMDAD (SEQ ID NO: 180) lO-niers ENNSPGIQLMKVSAMDADS (SEQ ID NO: 181) TVS IPENNS PGIQLMKVSANDADSGPNAC (SEQ ID NO: 182) 109PlD4 v.1, v.2 and v.3 SNP variants T486M and M491T 9-mmr NNSPGIQLMKVSATDAD (SEQ ID NO: 183) iO-mers ENNSPGIQLMKVSATDADS (SEQ ID NO: 184) TVSI PENNS PGIQLMKVSATDADS GPNAK (SEQ ID NO: 185) 109P D4 v.1, v.2 and v.3 SNP vajiants T486M and M491T and TVSI PENNS PGIQLMKVSATDADSGPNAE (SEQ ID NO: 186) 00 00 109P1D4 v.1, v.2 and v.3 SNP variants T486M and K500E C-i TVSIPENNSPGIQLKVSAIDADSGPNAE (SEQ ID NO: 187) 109P1D4 v.1, v.2 and v.3 SNP variants M491T 9-mers IQLTKVSATDADSGPNA (SEQ ID NO: 188) GIQLTKVSATDADSGPNAK (SEQ ID NO: 189) IS-mers ENNSPGIQLTKVSATDADSGPNAKINYLL (SEQ ID NO: 190) 109P1D4 v.1, v.2 and v.3 SNP variants M491T and T486M C 9-mers 00 IQLNKVSATDADSGPNA (SEQ ID NO: 191) GIQLNKVSATDADSGPNAK (SEQ ID NO: 192) ENNSPGIQLNKVSATDADSGPNAKINYLL (SEQ ID NO: 193) 109P1 D4 v.1, v.2 and v.3 SNP variants M491T and T486M and K500E GIQLNKVSATDADSGPNAE (SEQ ID NO: 194) ENNSPGIQLNKVSATDADSGPNAEINYLL (SEQ ID NO: 195) 109Pi D4 v.1, v.2 and v.3 SNP variants M491T and K500E i-mers ENNSPGIQLTKVSATDADSGPNAEINYLL (SEQ ID NO: 196) 109P1D4 v.1, v.2 and v.3 SNP variants K500E 9-mers DADSGPNAEINYLLGPD (SEQ ID NO: 197) MDADSGPNAEINYLLGPDA (SEQ ID NO: 198) TKVSAMDADSGPNAEINYLLGPDAPPEFS (SEQ ID NO: 199) 109P1D4 v,1, v.2 and v.3 SNP variants K500E and M49iT TDADSGPNAEINYLLGPDA (SEQ ID NO: 200) TKVSATDADSGPNAEINYLLGPDAPPEFS (SEQ ID NO: 201) 109P1D4 v.1, v.2 and v.3 SNP variants K500E and M491T and T486M MKVSATDADSGPNAEINYLLGPDAPPEFS (SEQ ID NO: 202) 109P1D4 v.1, v.2 and v.3 SNP variants K500E and T486M MKVSAMDADSGPNAEINYLLGPDAPPEFS (SEQ ID NO: 203) 109P1D4 v.1, v.2 and v.3 SNP variants 00 C517R 9-mets rN2~ APPEFSLDRRTGMLTVV (SEQ ID NO: 204) DAPPEFSLDRRTGMLTVVK (SEQ ID NO: 205) INYLLGPDAPPEFSLDRRTGMLTVVKKLDRE (SEQ ID NO: 206) 109PI04 vM, v.2 and v.3 SNP variants N576K 9Mers PVFTHNEYKFYVPENLP (SEQ ID NO: 207) SPVFTHNEYKFYVPENLPR (SEQ ID NO: 208) DQNDNSPVFTHNEYKFYVPENLPRHGTVG (SEQ ID NO: 209) 00~1 09PI1D4 v.1i, v.2 and v.3 SNP variants S678Y c-i 9-mers KPVFIVPPYNCSYELVLPS (SEQ ID NO: 210) lO-mers NKPVFIVPPYNCSYELVLPST (SEQ ID NO: 211) 1 VDVNDNKPVFIVPPYNCSYELVLPSTNPG (SEQ ID NO: 212) 109P1D4 v0, v.2 and v.3 SNP vadants S678Y and C680Y 9-mners KPVFIVPPYNYSYELVLPS (SEQ ID NO: 213) NKPVFIVPPYNYSYELVPST (SEQ ID NO: 214) VDVNDNKPVFIVPPYNYSYELVLPSTNPG (SEQ ID NO: 215) 109P1D4 v0, v.2 and v.3 SNP variants C680Y 9-mers VFIVPPSNYSYELVLPS (SEQ ID NO: 216) PVFIVPPSNYSYELVLPST (SEQ ID NO: 217) VNDNKPVFIVPPSNYSYELVLPSTNPGTV (SEQ ID NO: 218) 109P1D4 v.1, v.2 and v.3 SNP variants C680Y and S678Y 9-mets VFIVPPYNYSYELVLPS (SEQ ID NO: 219) PVFIVPPYNYSYELVLPST (SEQ ID NO: 220) VNDNKPVFIVPPYNYSYELVLPSTNPGTV (SEQ ID NO: 221) 109M1D4 v. 1, v.2 and v.3 SNP variants T7901 9-mets INELVRKSIEAPVTPNT (SEQ ID NO: 222) LINELVRKSIEAPVTPNTE (SEQ ID NO: 223) 154mets VTNATLINELVRKSIEAPVTPNTEIADVS (SEQ ID NO: 224) 00 109PlD4 v.1, v.2 and v.3 SNP variants K846M CI 9-mers HLKAAQKNMQNSEWATP (SEQ ID NO, 225) PHLKAAQKNMQNSEWATPN (SEQ ID NO: 226) RCRQAPI{LKAAQKNMQNSEWATPNPENRQ (SEQ ID NO: 227) 109PI D4 v. 1, v.2 and v.3 SNP variants F855V 9-mers CI SPKNLLLNVVTIEETKA (SEQ ID NO: 228) 1lO-mers HSPKNLLLNVVTIEETKAD (SEQ ID NO: 229) 00 KKKKKHSPKNLLLNVVTIEETKADDVDSD (SEQ ID NO: 230) IO9PlD4 v.1, v.2 and v.3 SNP variants c~K1 S958L 9-mers IQPETPLNLKHHIIQEL (SEQ ID NO: 231) 1 0-mers QIQPETPLNLKHHIIQEJP (SEQ ID NO: 232) 1 PQPAFQIQPETPLNLKHHIIQELPJDNTF (SEQ ID NO: 233) 109PI D4 v.1, v.2 and v.3 SNP variants K980N 9-niers FVACDSISNCSSSSSDP (SEQ ID NO: 234) 1 0-mars TFVACDSISNCSSSSSDPY (SEQ ID NO: 235) LPLDNTFVACDSISNCSSSSSDPYSVSDC (SEQ ID NO: 236) 00 00 Tables Vill -)MX: Table VII 0PD4v.1 Al- Each peptideIs a portion of1 SEQ ID NO: 3; each start Iposition is specified, the length of peptide Is 9 aino acids, and the end position forl each peptide is the start position plus eightj FI[l _61 DLEEQTmGK'I 96.666 189 LVIETPEGDK ,800 [FJ C94TFDPDYGR~J 1250 278 fGNAIF 11.250 [275]R1q G=ENAK 1.0 [FiIq 11. sGPNAK J[1Q9.-o DM~l FLSENIPLNT U .50] [8_[STNPqTVF 11. 5.000J- [67]4IVPPS NCSj I P13I 2iL DhP FYi 5.000 122011 KVEDGGfQ][500] 807 S DY V9LV3.0 591TMFKVI 2.500j SIDIRYV 2?.500 19_32 SDARY 2.5003 [78911 TEAPEVTPNJ[.25 .125 _.1EVIEN{.800 5187 DSDNRVr 'iI1.500 1[4_?9 IJ GIqLT IFT15001 9851spxy~i~o (991 rVSOCyPW1 1.5R7 F VTD,[gLGU- IR Y*_1.259 F273] GE 1.250 Fq-jf±! F 1.250 I Tale Vill 1O9PID4v.1 Al- 9Mers -Each peptide Is a portion of ISEQ ID NO: 3; each start position Is specified, the Ilength of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 521L1VVKKLDR 11 1.250 F1TTGAR1DR 1112 779f [J]UELVR =.250.

[2E9 ffTDqKMPQJ1.2 [jLPNSA[ Sj1.2 F59_ffLff!MRPrDY_ 11.00o LqR4[IDVSPT 11.00 972][ ffACDSISKIL.0 518 LGL 1000.o D85L4 t wENR [1000 QKEDREEK .966 FL 61j DRETPNK .900 I2[.L!EDNDA 0.j90 F98J11CSSDPYf 0.7501 FWL[KQESYFV 1LJ 172711 D(;TGNlT!F:6751 ji 61 TDVPLnIR II 9.TL57I 1612 FI NDDFTIDs j 0.62 ~804 il SPTSDYVI If 6 :1T [v -qtP6 Fdr~R 0.50 (io~li~uVQ Eol i 6jj 0 DE91 F 0.500 [§2J!P pysl 0_50011 [Table VIII 1 9PlD4.1 Al-] 9-mers; Each peptide is aportion of1 SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino 'acids, and the end positionfo each peptide is the start _p2osition pus eight. Io F895 ]DVSDGNRVI .0 EijI1 AEDGGVSH050 164251 ADEGI[so [E40] ArU I LO500] U17 JLIISTGY __VF.9O 3 18J6EPGT IL =.4501 LP09.jf VV Ilq 0.450 78-0]TIEVI .0 I25ffLY§!?.Ej4APVJ 0.300 [940 K=SQP 0.300 851 NSqEN~?J0.7 0.250] 6=6 [W _NKI'L .2501 .F38_7 IIKDADHGR JP2;; J K9 iLP§_N Vt I0.2501 Table IX- 109PID4v.1-

I

00 00 LEach peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 10 amino acids, and the end position for Leach pepfide is the start 'FJN-- LLETaAYL-DY (225-.0c 14k82 45~PNT (2.000 '(19511 YLDYeSTKEY lL2.o] 416 K NgGRVSRJ 18.0~cE 366 IVDTdYGD 1L!50 389 TID qTGV1R 10.0007O 1757 1S~SVD 7.500 12211 DNVPsIDIRY I~6.5 15751 VS S t~YVj6.00 F4Oj7Lo §-yvM ILS4001 445 FVpN .0 568-1 [~VP~I 4.7-1G 13081 LDEnDDqj? I 7 EWV-NdTGMNAL.500J 286 f S)CrgMQLT K =2.5001 11RL[DNiVP, 2500 1 4 76 [IDT G A V RYJ L2.500.J 2761 LLGPCIAPPE-J200 763 v~sD YPViT L P I751IE1) 2!:PT 111.350 115 Tq~dIGENA I 1.250 630U NPENrqlMMMIL I.IM5 2031 ETIeVE 1 11.25 T iF-54TIDADPoooI 515(p[G hRV=LVK 1.000 I TbleIX-1OP1D4v.1- All-ers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for, each peptide is the start posionpunne Efto]LLbsequencej~iscorej [47]=1A RTGM-K

OJ

A~tLo EVI !Lj 2 QY 0 1 DATNLnHPFj1.00 D A =DHnGRVTC gjEIEIEFsIPEAJ .0 F3NSPVITHNEY =.750 I1tSNvsiA =~.750i F PD-20]S I0.625- E331 E@DPVFTH{0E.6251 438V NDkPV FIV 1065 Igfg lk j _L~K OjI 0.500 M~j QlQfeTPLNSJ 0.5001 28q EAPte SD C ff. 1-oOI PL69F1-FTsK-EPJ1 0.500 !110I.CD~DLL i0.500O ~I4~il IIGN]DO !I0.00 L721 Ks~qsPj[E 00; 15501NATi5NELVR Fll5-M-11 F Table IX- 1O9P1D4v.1- AI-lO-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position forl each peptide Is the start position plus nine.

F83L[ KE PIDR ET =9A.4q 28l VSIPeN APVG 10.3001 [6E]&ADW, SGNR02 Pl21FJILQ-saMLR-K5LO2S] R144 NDgM NAEV (530I PDS IF§VVI Fl-.250] [23]3 0.250 FL6 IFLE IyG1~L0Z25Ej lt33EE :=5YLFI =0.225 351 VPENIPRHGT ff225 I 21 TEv aKLA 0.2 EkflLW FAIYRi 0.200 JYLsNfGT (L?21] STable X- 1O9P1D4v.1- 1 .A0201-9- nrs.

Each peptide Is a portion of SEQ ID NO, 3; each start position is specified, the length of peptide, Is 9 amino ,acids, and the end positIon for each peptide is the start position plus eight.

FoEi jLS ue n ce s i IFjLS FIITAAYL 8198.910 F541IIkP-DElFRj[516.2721 -7 00 00 Tablex- 109P1D4v.1- A0201-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 9 amino i acids, and the end position fori each peptide is the start F6]71 GQPDSLFSj 385.691 Ef7]GLMPARAM jy ~A I I F 9: NNVTI 73.343 [6 jRVIRFLI.. 60.510 ILD1FENDT I 5.992.J 598 j KVT DJ48.9911j [23] NIRRFH L39.16 47-91TLKD 35.385 FTO 4ESvv:lN vNLUJ 3.=7 F4KLYKG1 31.646 FIQTrMGKYNW,[ 29487 F17AL9Iq rJL40.117 J 75-3]EILVAGTIL29.137 1 NTr-yI 28.690 23-8 RLHN~ 7572 [121SQNFGLV j26.797 930JSVSOCGYP 2 4.5 E74 EKCD 2271 ~I7 IIFNSNA 1.219 ![jTMGK 7 NWV 1-6.55 931LwyvI 14 V VF .654 F 2] NVTDVNDN F-1i VVLSE-N PL II 1-1. 757-1 rTable X- 1O9P1D4v.1- A0201-9-mersJ Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start I _pos~ Se uenc hLSc QFKLVYK 10.93 E[99 L vPINoj IL84j 247 LMGARAKEPI 10.746 21-01 ~433] I490ILTSVTV~jL952 1) 1 RV=KADL L E hNEYNFYI6.1

SNPGTVVFQI

6]3 v 60 5 7

J

F7-5] AVAGTffTVVJ.3 VV7F66] =4.242] 116 IESN Lj[ P3 ~F4VLmz7! 17§?JLEYWl=l lLIj 67 F i90.I..I T EVS F -2.911 T] APVFQSF 2.497.

AFS .WCRTG 2L.263 662FmDLFAiDQET I6 ~Tabe X- 109PD40.- A0201-9-mers Each peptide is a portion of SEQ ID NOr. 3; each start position is specified, the length of peptide Is 9 amino acids, and the end position foi each peptide is the start position plus eight 4PlosjL Sequence] Score- I!8A5J[i:ijL2000 !F[3521 FN FLL E956 q35 j[NaFLLrA-A[ -1864 89Jf~~ 1.85 P275. MPARAMVLJ .7 1266 MP9L 1.2 F!L00_ vELN iLI.195 D~kF -SENI( 1.163lI -A2r1 DKDAi116 [77] RQAPHLJ 1.159_ FI913 FEC=DSISKC1LI9 27VLASDGGL f 7 37-0 KYAKA 1.8 M120 [SPGI-QLTIKY I [T044l EI71IISATLQ-v-sv]F-T1 06 Eq6jJTRPvGI L. 0.913J 658GTDFA .0 i35OIIPVFSNQFLL 1 0.882 1 00 A0201-9-mers Each peptide is a portion of SEQ ID NO. 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start FEo-sliu] LSore 314]IPL NTKIAL [0C.877: [Table AI 109PI D4v. I-A0201 1 10-mers Each peptide Is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is amino adids, and the end position for each peptide is the start position plus nine I 274E PNAMYj- 196 4071 701 [J:SvvVN~j181794J IJAJPdEJ J144.981 223j (fsFSYLAV ll1273 Pii jiis34 [K7] MaRMLL3A7 L[2:9f IVNpV@TVJLI 2.56 WI2 NLrhIV 1I362I 176.1 T!TVvYfl .118.147- [§-iTLLNFvTiEET P114.2771 f NQAT!LFI I~ 13398 [L-Tfl FVNE'sVrNAT !j 12.298 [ii] FSVVLFV I 1.4871 F6-'6 LG=SFS 0.2981 Table~~ X-1OPD4v. 1-A0201 _____lO-ers Each peptide Is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 10 amino adds, and the end position for each peptide Is the start position plus nine.

P~ ubslr -eence =~r [6721 N7rLxEKCPvjI956 D1731L ILWvS-V 8I.72 Y NFGIDI 8.720 93-41[CGYPvnMY. Ij847 4891LLT n3Y i 16i F21 i QEIP LDNT j8 04 =936YE tfVP j =7.936I 1=45KL =EKTI7.693I LDO iLGM:DevR-sl j[j.

f~ =~[TPtFV[6.057 126 KSQM4FGLD 1608 402 LN:PVf QSFV 707 FVNUVEV[~9 321 AIvDDAJ49 11F4.:24] 478 (478][y IZ E23] LV1IARRL IAh F62]J L VLPsTN PGT 4.0 6385 FIMEaVDD 3.46 Tiable Xl 1O9P1D4v.1-A0201- 1O-niers Each peplide Is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acdds, and the end position for each peptide Is the start psition plus nine., [4-5j] SLDrT L =2.98'i 3130lnn1[2 10 91 EIG NqvQN YLJI 937 I [~fF=903j 1[7j 5=7 nLSN ]=2.495j ,0370JKYILA E, 1781 R~i 2.31 F(--9[YSVSdCGYP =2.0§8 7fl[ LAAaGK PPL1 _2.0681 M Nl 1.953 =2~~Ll 1.869 [307 IIJW NIPL =1869] 233]SIrLH .6 43-51SPaIY .6 Le9A1 ILW~kVFIV. .11.689] 2-721 GG~AAV 1.680 181 IIKLUNWy EI.[467 193011 SVeS~cV =.6442 [ijIVA~aT 1j.642 q_ 1.578.1 .y104 i VVDNVJ =j.549J FL Tip ,KPVJj 1.5449 Wi =M~S~~[1435 NI-E8A .41351 00 00 V Table XII 109PID4v.1- LA3-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is The start _position plus eight -Pos LSubsequence Iscore F-751 90.0001 r(W-1J DLEEQTMGK 18.000 F803 QMIMMKKKK 15.000 781 JLKAKNK =10O.W Di]D7..GurniKPL IF4.050~ ILPFEIF 7 500J L66=3 ]VRNS 2.700 I387j NQSALFI J[ioo 4=4Lr~ 2.2-50 I F767-1 VIFrTAwR T -66-6-1 F590-1 RVSRSSSAKIM Li F 8-11 KTGDVPLIR 18600 53- AIPDIF 8[ i00 mimmKKK GLHRVL11VKA 11.3501 274j'1 i LMAAMLj1.200 4-58 j RTPMLTWK 1FO0 129 V1ETPG 9K [061 855J1 Th0-901 Table XII 109P1 D4v.1 A3-9-mers Each peptide Is a portion of SEQ ID NO: 3; each start position is specified, the length of pepfide Is 9 amino acids, and the end position for each peptide is the start I posito lubseigh P Ti-i -IT I O F--2-1 IAI1VD 0.900 [iI L.-R v0 O00 =802-f StIFIKV 06751 j~ I1 J I F719- LATLINELVR II 61=4 VPSCSY ][E00j LTWKKLDR I]=O 0 709 L 1- -i1~so =~LL J0.000 F4-9-3 IAGT 10.! 0 895761 QETn P .540 [i41 '11KPSPD~ 0o.3601 F05.3TH6!0F 54 ILENDF W0 18_21 i LLLNFVTI-E -0.270 F11 KLVWKGDVF -70 Table XII IO09P1 D4v.1 A3-9-mers Each peptide is a portion of SEQ ID) NO: 3; each start position I specified, the length of pePtide s9 amino acids, and the end position for each peptide Is the start position plus eight.

Poe LS euence I Score [T7-1L- YyMlEA F10.-270 E jii' NIRR FHL 1.270 7D IL KIRFLIEDI f10.270 680 -i i TDGLH [.-240 F4i7i11 YTLMEKCV iI 7662I1 DLFADE J~02-25 787-1SPDW HYK-1 200i 7775JLC L LK 2O1 r..TI LFtHEYNff j0200] FL84~ R K J[Q0.200 =633 -DCTG7 110.180 KVTINYIIJ0180 [4=2 E[DVSSPTSDYj[ 0180 [2-4jTj HLNATTGLI_110.180 [3-1 KQ Y F O.V 1-02 [l9i5-1 NTFVACDSI 11-.150 62=8 7G 1vFO50 FL Table XII 109P1D4v.1 -A3-J lO-mers .1j Each peptide is a portion of SEQ 1 ID NO: 3; each start position is specified, the length of peptide i amino acids, and the end position for each peptide Is the s__tat psi!i Fftosli..4bserice Score 16831DGiVV 36.000 [319 IKAV1K E501J 1~rTPj18.000 00 00 ITable XIII 109PlD4v.1-A3- 1O-mers jEach peptide is a portion of SEQ ID NO: 3; each start position is jspecified, the length of peptide Is amino adds, and the and Iposition for each peptide Is the L sart posit plus nine.

07[s I Subsequence LnEs-yi Iri6. 9p 1115.000 14@]l QLIVqKELDR ~Z0 E467l1ALPReKEDKYJ[2.00 F347 MR KKKKKF 1.000 Fr~ffGMNeRys, F8. 100-1 461IMLTVvKKLDR =8_00 160 KVEDgGfflR ;I !.60- WI AY~dE~3-.00O] jRRI LLGdAPPEF 1 [N7L--pn J2 2700 ED7 PLF~aTVII !L7001 L VIRPnISFDR_1L .7 89-0] IQPEtPLNSK ]2.025- =766 I GTIrvvvvIF 205 [F3831 YLDYeSTKEY 112.0 1~Z~LLM~cD~DL] 1.800- F57 LPDEIFRLVK .0 WL9-1GI~EHCFY ]1T2001 63 I 7 ftiV!pSNC =0.900 ,5~F11FrJVKiRF [P900.

t-6j GINjvoEL.Ky 0.810 124811 UTIkEPLDR I10.866 FEi~]I FI~dNN I 0.675 Table XIII 109P1D4v.1-A3-J 10-mlersI Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end 1position for each peptide is the start position Plus nine.

FPosl Subse uenceI.j LiIAILPdEIFRL [o.Los] i 7 18 1- Tyy-JI .600j F5272 5L.:TG I 600] F3[-4 5 UEM AY 0.600W [8=0]RQMIm=MKKKK 0. =5 5-2-31 IFGIDV IET =450so ff51F EVss tsVj =0.40 L5-57] DT vPI 7S _{o.400] !11~LV-GlnGVQN ~j 6 [111 LVY NtGDVPL ii 0.300] FiijflDiT:nHPVK

O.

Ff5[IVGkLDREK .300] 5 D2IENPMavoPDV .0.3001 f4 LDGL ~k 0A7 MjV F9SAMFIK I 0.35j Sal II- 1O9PID4v.1-A3lO-mers Each peptide Is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide Is 10 amino adds, and the end position for each peptide is the 1, s tart position plus nine.- 7 LTcF Rtd IP o .200i LD~TGML IL.2001 L38-jJ[LQSaM LFIK rI 148 NSgILT j0.180] D 11 N GV LIK Lii FgP-LaPLdNTFV-A 0. 1= =0.180 II AqF 0.1N80] ,j 866]ETT=FK DSPDL[0.01 '1Ai[ LLNFv11 j[o AIJ TvT KDA IL F -0135 L ~Is~R~ =01351 [R JL _YVIFTAV J0.135 1Table XIV-109P1 D4v.1-AI 101n 9-mers Each peptide Is a portion of SEQ ID NO. 3; each start position Is specified, the length of peptide Is 9 amino adids, and the end position for each peptide is the start position plus; 475 ]KY FTIA -3.6001 45 (rMTw riK 00 00 TableAXV-1O9P1D4.1-A1 101- 9-mers Each pepfide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 9 amino acids, and Ithe end position for each peptide Is the start position plus eight j S]fubseqence j cr I[lJ[IffYDVNDK II 3.000 1 LF80 NQAMFI2( t]0 1=37J _LRKD =.200 =8[TGVL~ =1.2001 A4-f T--G-LTIKf 1.6000 F720 -][1N=ELVRK F-~o7 F1?49?j[ ITIKEPLOR] ]f 0.6001- I 77-5 1 R0.AHL] 6=00 7 19j [ATLI NELVR =0.600j 362 1AYLDYESTK[O60 =W3 I MMMKKj[] 824 LFKMMEETK L9E400j 5-5i1 PJEIFR II 0240 3A 1841 LYRSQKED I1[i] SAMUIKVKj =0.2400 J9I 1I SDLAR~ 0.20 [D 79AHK K *0.20 I PTLISj.20 ~J TOHIPr0.-2001 Table XIV-109PW1 D1-A 1- 9-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide Is 9 amino acids, and the end position for each peptide Is the start position plus eight.

1F!Eos1subsee Score 4=64I WKKDREKJ 0.20 Lz~JI~u:r Jvv =0.160 5=6I QEs-y-Th2S][9A .L [~]Evvs~r 0.120 68-8-1[ R=VAL[0-090 _31= SENPLNTKU[66 F41W Irr =0.060- I704ID DTGMNAEVR[ .60] F4327 ED DA PNAK I 0.060 F-LfT VTFQIA 0.060 6= EKf jj) KVEDG 0060 F6v971j =0.050] Izilli GEIFGAR((O054] =0.04 k~~f GIL EKf000 i182 TN DNHPVF-K 10.040 I ATPNPNRI10.040 Tbe XIV-109P1D4v.1-A11O1- I 9-mers Each peptide Is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each petide Is the start position plus folfs( Sbse quencej Soe] 4 59 ILTqMvLWMKJ .4 152- KDTYVMKVK 1~ 0.030]l 766]WFTW I000 I 280i IIwmVLNV 6(0536 26=] LVADG]0.03 0 I 6] TWFI 0.030] F30JI IRKLAJ 0.024] (273 GM fAvVj0.024] Fj0:JI ENRQ-MMMK If 0.024 L-11. RQAPHl .2 L SFVHATRPVI =0.020 750 YVILVMV 0.020 Table XV 109131D4v.1 AlI101-1lO-mers Each peptide is a portion of SEQI ID NO: 3; each start position Is specified, the length of peptide Is 10 anmino acids, and the end position for each peptide is the start position plus nine..

11 Po s ubseqce Score- 7 19 ATLFnLVRK 00 i 00 00 T-a-ble XV- 109PI D4v.1 Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide Is amino acids, and the end Iposition for each peptide is the start. position plus nine.

ubs e LL uence Score F wLKVyEDgGFPQR I im 1128 IDV IE WPEG-DK 090 1Z6!.. f[VFITAVVR j[ 0600 [69j ETGNMUMEK 0q.600 1I43 IVQKID REEKJ060 [K6] FQ~PNIJ 0.600 J] [E[fMq QKK] 0.400] [=61 LIMMEkKKK] 0j.4001 1[61 L Dff L[ 0.4003 i[I~JJ~mnH~v~jD=.300 463p1l KKDEjI 0.300] 4 F R~ 0.30 If54 IR ISFD I0240 5D !LYS vG N j 0.40j L~JL~4yi 0240 6IL:G VAKf 0.200 8=06~ N i'l 0.200 21439 NATT 8TK_ 0.2 1 F247 T-10.160 [3-6I LNQSaMLFIK 0.120 27A I ?F.dLD 10.120 1 524 j VAI@pDEIFRL.120 I[ qi[ KKLDrEKED1 0.0960J TYVIAV I 0090J [718 NA NLR 0.0 241 IFTgA RIDR I i F Table XV 109P1 D4v.1 I- All10-lOMers Each peplide is a portion of SEQ ID NO: 3; each start position is specified, the length of pepfide Is 10 amino acids, and the end position for each peptide Is the sI-_.tart position plus nine.

I PosTSubsequence[ 1 111 N l~GVliK 0.060 849 LILRM~ .060j [Ljfl[wwdvtD Kj 0.060 FI1-li GV E I 81= i YESPK L0O: 1 E5UE~cvv-PLI 0.060] 1.760 3i GTIDvVVFWI 04 [LF9:s0tYVKJISAK 0.060] 76=0 CFThIPFR 0.0405 VtPWILO= [Zi1[]viFAv .11 0.030 J4- E[C, FLVLASDGGL][ 0.03 ]LE nP-h IPF 0,040- '(34-1~V~NFL~ 0] I~QYE~ .2 F51fl MDFsPNK 000 Pqi 214j DA frEAJ L:I63 Tbe XV 109P1 D4v.1- All0l-1O-mers Each peptide is a portion of SEQ 10 NO: 3; each start position Is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine.

o ~Subsequence cr 7570 A VNAvNtW 0.020 Z74j]VRC qPLl0.020 I Zo7J ILLIvNES-vJ 0.020 S750 F I YIVVA, 0.0201 255flDRePH .2 ET- wkD

PDL

[3j[SVT(- CHATA 0.020] S93911 E ySyHJL0.0 45=7[CRTGnLTVVK =0.020 725 sTEPV L f6551] IVG-GnTRDLF i,1_0.020 P3 I~VVRQffLI 0020 !K~sFVV' 0.018] 55@IDQG V 0.015le XI091 f DffV.1A2 KI-[AI~ 9-rs 08 ofpeti e s m noadsan tedposition f eth peptide is t start position plus 1 F4 F± L paoooj [iVYKTGL ~2on~~ F3-4PFSNQ1 14.400 00 00 Table XVI-109P104v.l-A24 Each peptide Is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus.

Pos L-i Lc ece[ r-t 59 K] SYEVPT H I 102.01 [=21]_§YEI v l1.50 6 RHNEYNFf1001 [2=23 KIHFSFSNLI9.01 i V~EP l 8.400 F514 EN FYVPEN8.5 67=8 KC DVTQLGL rj 8.000] =jGPN'=tI .200 LIL=4 IR .200 717 I2TINELJ6.3 667 F2 -2E T GN IJ 6.00 [274 =MA~tI 6.000 [417] [N W GIL E 000] 314 =PNXI 6.000 1302 NPVNT =6.000] L U IjSNYIL 11 600 WffPNHMkLVL I~q6.000 Lgi]1F=QIQPETPL I F27ISFSNLVSNI iF 6.00 1 F231 iLSIRL 5.0 148 Table XV-109P1D4v.1-A24 9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position plus eight Fqsji !!bseguencel[oej 7i1If IN GQYELII E To J EKL][4.400j 61LRLVKIRFLI =4.200] 37= FAD GKPPL1 .0 88=0 KSASPQPAF [IV GNTRDL I SAVRFL 4T0=00 618 ]SCYLLIA 752] LSIj1 4.0001 5231 LPRHGTVGL 4000] 44=5I RI E 1=jL3960j Sfl DND] =3.600 11:7]ELISQNIFI360 605 NAVFQS 3.600 1 T DNI-113W .600 211 D INDNAI.F .6001 6 2811 F*PGrTV] =3.600 52JE YILPDEULaOOO I -iU GVIRP NISF FI 3000 95=3 NIRLF 3.0 212=0 9N!U :9 76= IGQVNT 1 3.0 !I Table XVI-109P1D4v.1-A24 9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptIde is 9 amino acids, and the end position for each peptide Is the start position plus IF[osjL6 biequencnJI-cre-J' .817 I SKNLLLtFIL2.4001 [3121EILTI L 762f [jITVVVnIFj 2.100 695 IDLGQPDsvLFILw-01 93-3] DCGYPV1TF2.0 j86 j ISPENSAI EL~ 306 DTWLSENI 287 Loy!NPNVPSI I±~ 1-62 E!MvDPpyGI 180 ii 647 I MNAEVRY SI L FPQRSTA =.600 Table XVII 109PI D4v.1 A24 IlO-mers

I

Each peptide is a portion of SEQ ID NO: 3; each start positWon Is specified, the length of peptide is 10 amino acids, and the end position for each peptide Is the start Los ipus nine.

bsequence JI cr ~514 INFXYEN L 000 LA I f O yL~I yEKqg I l FeL87 W DPI 887l AQqETPL j 00 00 Table XVII 10931D4v.1- A24 10-mars Each pepfide Is a portion of SEQ ID NO: 3; eadh start position Is specified, the length of peptide, is amino acids, and the end position for each peptide Is the start position plus nine.

PoS subs euencei Scr 23911[LFHLnATTGLI 20.000 F59-1 IF-RLVFIR-FL iL 2.000 702]LFSWIVNLF Jj i800 Pr]KPPnQ L 120900 8042LV=TLdLPIDL J .0 1716 ILrNAItLINEL iL .Nj- GMUVtWKI91L F-.i4 i111SYEvPSTN1[ 9.000 1749 FDWKGLVMV][- F 9!.000_J- 1024l T _GLIIIKEPL] 8.0 436 GFN INYL L.4 D165j FPTqrSSTJ 7.500 I 897 LsKHh-IIQEI, 7.392 [j~JRIE~TGEI J[ .200 ii-1I AILUdEIFRLJ7.o 14035 sGPNaKIY1 L =7.2 f2-7-3] =L~RML .200 1~]IF LGMLI .0 8615VPnSE .0 10-9 =6NvQNE 600~ 31-31NPnKA .0 [878]RKS8LSPQPAL 6.000 17121 VNESJvTLI .0 ,IPIi:RhGTVGLf F6.000 j F6.000q 11675 1 LMEKcDV TDr600 I r 202F G!SVT9LJ[o6.(00 F233 SN*~R LFHL 6.000 301 IVPDTY47F90:7 Table XVII 1O9P1D4v.1- A24 10G-mers Each peplide is a portion of SEQ ID NO, 3; each start position Is specified, the length of peptide is 10 amino acids, and the end *position for each peptide is the sta.yosIon plus nine.

Pos iFSub-seque-nce Score I .00 132TPE~d KMPQL600 55Aj sIV-G g NT-R-DLjj 6.000 347 N[ 5J6_ 701 I LFSMVIVNK6[T-Q 481 [LKn G V PP LI 80 13771IA~GPPI 681 [VTD gLHRVL I .0 k0zJI .ESTKYqI!KLJ FiA.4L 90 [L D Ln TFiiFA .9 I?- Fw--1IE!rEP -SPDL 4. 000 O 11J811 FYKSq@lFGLI [~JI~D~gQPSEL..000 1 M E jDNAvTLSL 1[ 400 5-1 LV-YK tGD VPL[4.0 DEG~DHC .0 L zJI LSB..dENDD)Ff ODO~0 D952IYGIQvSNUFII3.0 562, TG~r-PNISF II 3.00.

41 IDNAPYFTOSFL -?88o.

-5 '1I. EIJVKIRF ]IF2.800 49j TNtf VS j .52 217]DIr nKIF I ,0 87L IDvPLIRI F7j 2400 Table XVII 1O9P1D4v.1- A24 Each pepfide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide Is the ilstart position plus nine._._ P05 I F subsequence 1[ Scre [47'511 KLtILAKD F-2.31071 796 LfrNPeN o9v]L- 2.160] L NVRYSlI210 630 I N _PGTvVFQV1j =2.QIJ 655 LKGGnTDLJ E2.(00 F3fl _TCF 859 LY t EE0L .00 U31 F11DsQG"R oo 165 I NnK F L .800]O: F85] FAUy~qETN ILY9L F=IEDnNGI 1.8001 17731 II PiI [:650_ I =1.650 Table XVIII 10913D1M.1-137 9Mers Each peptide Is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus eight.

FP1 Su eo LIL??2 Ti jF~ 128 .F GAIREL 180 00 I4 .iI GPNAXINYL I 000 if 260I iNH~lA: 80.000] Fl3.211 NPVNDTVVL IF§a~o [732AVPTI3.0 7261D APLFPTnm[9.

00 00 TbeXII- 1O9PD4.1-B 1 Each peptide is a portion of SEQ ID NO, 3; each start position is specified, the length of peptide Is 9 amino acids, and the end position for eac petie is the start psition Plus eight [P-jsSubsequence 1[ cor 796 ITPNPENMJI 2.000 pq[1VLVKNDL 20.000 231 II LV IA RRLJ2.0J 9§AM QSAM§fI l[26=6 DGL[2.00 W iAPFQSFyV 2.0 3-7-8DGKPPL LlO.800 166 I QRSS§TAI 8.

745SPTD YVKI 0001 341PPLNQSAML 8.001 186 I RKEEI[ 00 ff"][JLTPLNSKHHL I616ESN-CSYELJ =8.000 FAi EFQIQPEFLj6.000I 449 APFL .0 41-71 NNSG IQ! 600 798] NPNQI 6.000J I 102 JLVqPDVGI ~50 D63 0 TNPNTEIADV [4.000 2751 MPRMVV 4.000j 46-0GMLTWKKL [=.0001 L4z v.I £.009j 167 I PQRSSTAIL- =I4.000] T~i LDEIFRI I 420lF QLJ 4.000] F6-41 KIR!FLIIEDI j4.0 Table XVIII 109PID401B71 9-mets Each peptide Is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position pls elghtj E~Fui Sue c oe I 62RILPsTNpGT-vII400 5421 NSAVTLLJI4.000 [RI]PROEHO: r515jEYRFYPENLj 4-.000] 7-17 LT!TINEL]4.000 10 Kv--SLC- F10001 F97076YI UNAVAGiVL3OJ F 2.000 2=96. rIVP 2.000-1 [87DNNYSJ 2l.0-00 1 [J LVMA.VA9TIJ I49~F i~iI 2.000 948 [Ht!PV-G IQj_.

847 _PDEQ =2.000I D882 AS§PQPAFQIII 1=.800 1[7i5611 mYA-IT 91 l2-7?lIGd[MPA Mi 1.5700 J[45] SLTGM Il poj 243] NATTGLrFI ?0 I [F]DPV05GVI .2001 698QPDSLSW F1.20-01 Table XVIII -109P1D4171 Each peptide is a portion of SEQ ID NO:* 3; each start position is specified, the length of peptide Is 9 amino acids, and the and position for each peptide Is the start L positionplus eight.

7661[qiLq T =PEFRV.2001 28=0 M IVNVTDV] I 757 WiFIA-TA J=.000I 93=0 SVS CGYPV[4J 7300 IVN1.000 398! FKVTSNVL__ jj qL=MPARAMV 27=8 LVNVE II Table XIX 109 P1 M. 1-137 lO-mers Each peptide Is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 10 amino acids, and the and position for each peptide Is the start position plus nine.

pSubsequence]I cr FAPVGtSVTQL!;iW ;7-77![WVVRCTQAPHL!. 200.001 .IGPN LidNYLLD1~00 5JPVsN23]18.00 00 00 Table XIX- 1091M4v.1-137 !!O-mers I Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 10 amino Iadds, and te end position for Ieach peptideis the start -position plus nine.

-P-OA Subsequence]I Score KPPLnQSAML L80=00 16-6]FQsTL 446 GD PFL3.0 13 TEGdimp!1 24.000 842 RTdPDL 2.0 7] TVsNP0.0001 W PIIEQT 20.000 Kv'T-yiIqDFF C2000.] '148! LAnGVPPL 12.000] LS!JI JdEffRL P=12000] 1377]LLAA aGKPPL jjI2900i LEach peptide Is a portion of1 SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the startj position plus eight.

ofts =Subequence ~Sore1 FEPRIEH'II36.00 ~~PRH=TVGL 1817 PNLLF16Q0001 I507T S PVFHNE Y: 4 -O-000 IFIIRYFN l171I 40.000 L87!INHLtsP Ljlap I 45-31FLCTM 2.0 7436J1_GPNAKINYLI LOP_00 Table XX- 1O9P1D4v.1- B3501-9-mers Each peptide Is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 9 amino adds, and the end position for Ieach peptide Is the start position plus eight L3441 IPFRLPVF~I~00 Pi4lISPTSTy!ij[ 2.0001 I2092~ FVPSIDIRYII 200 F6 39]_IA;DN DTG M_120001 P [SASPQPA7J1.0 E92 KVVDG 9.000 8094 y~~i =8.000j 186 HKfPV1F .0 00 F1661 FPQSSTA j8.000 ~J2 VSNIAR RLF 1500 542 NSAVT0S L '630 N-s~PGV V =4000 L61901 KPVF I VPPS 4. 00 0 D593I y1 J. 4.000\T- D44 =PILTVI4.000 847 L PIDLEQT [T209 ,275,, MPARAMVLV! F4.0 007 54-jJLPEDEIF [EO00j I Z NS!KTL I F3.0O- 5 101f FtN EYN E3. LK 591 VSRSSAKVI 3.00 0 RE]2 NAPFQSF" L I] Table XI I09P1 D4v.l- 833501-9-mars Each pepfide is a portion of SEQ ID N0. 3; each start position is specified, the length of peptide Is 9 amino acids, and the end position foi each peplide is the start position plus eight.

E[~sJ Fsubsequence]L core 'E548]LEN=DDF ]L3.00 502[ JILPD-F ,D K R*2.I IE2.PG] 2A00 RIflIIE EIR 1 2.EI 400 I55IL: _DEFRLV[240 LiO2 L VirpVq! =[2409L 2]JLKIHFFSNL][ 000 E72]FVSPT-j 200 13561 FLLETAYL I[.0 ~3 L2DIDL 2 E88]2AS QPAFQI F 2.000 qIM16 LPPNSE[ 2000 7F1 IESVATL 120 gEL IDQNDPFlI2.000 53]1 [-TUVDD I 2000 EWFsIMN YR 2--000 I~4~i L P F!I 2.000 4P64j Tm1FKPDSI1: 6:6 F68- IRT 2.000- 7119811VGINGVqNY 200 F!in[ NP-ff] E2SAINK IR147FLD-ii 1T I1.800 00 00 Tle XX- I O9PID4v.1- B!13501-9-mers Each peptide is a portion of SEQ ID Na. 3; each start position is specified, the length of peptide Is 9 amnino acids, and the end position for each peptide is the start -position plus eight 1Ej[]Su-bweuencjore iI47]0lE KEDKYjJ1.0 D-ENDNA1 .0 .3 DRyrL tKUA500J I903 [SSSDPYSV 1.500 [308]EWLENIPLJ =.500j F918 1.i 1.500] 39 DIREO 1.500 196- FI-R VSIPENAVY 1.500O 1 TGLITI[120 tr be X 109P1 D4v.1 Each peptide is a portion of SEQ ID NO: 3; each staut position is specified, the length of peptide is 9 amnIno adds, and the end position for each peptide is the start Itio[n plus eight LPR~TVGL 2400 9 00 Z]6GPNARINYLL_120 13] mpQuvQKL%0 FTable XXI 109P1 D4v.1I 83501-10-mners Each peptide Is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight [745 FPTYVKIL L16J ff~SAIJ2oo HL~20L1~001 jFl 87 NSKHhIIQEL 150 NPENIQIMM J12.001 DSG~nKINY10.00! [50611_NSPVTHNEY 1000J 63 NPGTVVFQV 1181 E~ITLNsKHHI!J 8.000 1547 LSIWENDDF 7.500 TKEyAlLLj 6000 F461GPO PEFSL =6.0001 F-2 TjPEGdKMPQ-:L 11:9-00 1F16I[ HS-PK-nLLL NF 7.90 El iI NS in lTj~ r9-2 KCS:SDPY19JF Table XXI 10911D4v.1 B3501-10-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide Is 9 amnino acids, and the end position for each peptide Is the start _posltion plusj eiht 6101UL \FLPP§NjiRIo F487 I FVPL t N-VTVI4000 626' 4LP.nG 6wI qoo F906Lq F=VAC [0:00] 6064i [7-4]S =PTsDYVKI [3.000 1D18 LKNFL[~ 266 [LVLAs=GGLM ERI~ 7795 I-98i Q PRSIFSWI~ =2.400J 226]LFESu7 LVSNI JIO999 61311 FI SNCSY DMEPLFpTVN =00 945 EI VSV _PG j F 530 JLT-vTPDY =2.000 1107! V \Ts-pRl]Pz99I 172-811 KTEaPVPNI(.-M [302 EN dTW7 L-LS-JF? 7 0 FWTGnAVR 21 E nAKIHF 00 00 T~able XXI 109P104v.1 B3501-10-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start 107 psLbon pIlus eight j P~LVALoo IF73-51 PV 12.0001 491 TSNVtVFVSI F -0 I~~I PL~sVT 2.000 KDv PLIRI 1.7600 F53 I~EFR ~o F307 L sENIPLIi I 1785 LF9KkNSE I&I~ L22] SSDPYISYF5-0 JF91 Y IPVnDTWLj iIl50 "5911 Il 5Y-iL50I FVFI DREKL 1.500 4Oi] 0IRDeHqFY I[1.001 69-2E] KADIGQPDS 1200 171-6 I REdTGEIF 11.20 [~Z~IREK~eYTF =1.2001 LTable IX- IO9P1D4v.1 Al-i 0-mers Each peptide Is a portion of SEQ ID NO: 3; ac start position Is speciie, the length of peptide Is 10 amino acids, Iand the end position for each peptide is the start position plusJ nine.

F~ols n Subs LLETaAYLDY j7. 0[ Table IX 109PI D4v.1I Al-i 0-mars Each peptide Is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 10 amino acids, and the end position for each .peptie is the start position plus I nine.

DLEqTMGKY i[o [43 3J] LSENiPLNTI(]270 -1 1 103]J AVJdGJ j2 160f KEDgGFQ 1 8 001I KAEDgGRVSFR8.5Y IS ASOGgIMPAUR=lf iioo LPDEiFRLVK i 25 5 IL H~ D6p-[.2.50 t~IL DNSD j 620] _]43 VISSqTGDVK IT~~ KQESyTFYVK _i5-40-j 6103 FIVPpSNCSY- 5.000 l IENsAINSK_ P.5001 TSDfvK ILVA, 1 3.750: F4 _NNSPgIQlLTK. 11 :146' I EL-DReEKDTY 2500 IEnDD I E644 SLTG !4Lj2.II 285 VTDVnDNVPS7 250 Table IX 109PI D4v.1I Al-i 0-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 10 amino acids, and the end position for each peptide is the start position plus nine. f~IlAVDdTGNAF 2.5001 193-111 VSDCgYPVT1T W 1213 ATI)dIGENA 15.25 RN9 IT~DHY 1.25-01 F8 iIvTDLgL-RvL- I1l.25-j F7-1 NPENrQMIMM ,1 1251 ~15-91_L ETEMevSIPE =.1251 D1811 DjTNqnHPF i0 F37-81 ALD~iETPEGN 1.000 1683 DLGhRLKI1 .000] F2151 I gMJTWKItOOgj .719 ATLInELVRK 1. 000 1 0530[L GLrvTDPDY J]~ 329 DADHnGRVTC 1.000J I[8:1 TIEEUADDV ][0.90 1i903l ELEVslPENj 0j 5006L4PMVTHNEY 0f.750i Fqq__ NDNkPVFIV_ -10.6q251 P l-9 1 ENDNaPVFTQ- 0P.2i IND] WNaPlFP A fP.6251 F§23. QNjsVFH 10625] 1P-611 qAC ESIS I 0.625 lE781 =C~DGH 0.500 718 NATNLV ojF000 00 00 [Table IX 1O9P1D4v. 1 Al-1Omr Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 10 amino acids, and the end position for each !Ipeptide is the start position Plus: nine.

1153 Id FRL][0.5 1248 1q:50-E-DR 07".0 1430 AMDMdSGPNA7F- ]6:-o 832 0.DVSDN]50 1273 L~RAy 0.5001 VIRPnISFDR7 0.500 [3371[ TCFTdHEPf_][Q-i 1 1[LIIDWDNSPV 10.500[ 448 I DPFOp 05001 1140 1 FO ELD 0500 I1071I DVGInGVQNY] 0.500 __-601 GTi vV Fo -[.500-6 f19201 KCSSSSDPY =0.500~ F8-1] SASqPAFQI 10.5001 VDNPVf 0.5001 F2-61 TTGAfIDREK [0.501 )~DT FO.RV 415-0 721 Y~SVN~ 0.450 196 LXSIPeNAPVG_ DO0J *lejI .9SAYIIKYKII..00 N1811.- 1 DDIARHYK jEo.3-j .161 ISIe NAIN[0.00[ [83[ 5ADDYdSDGN] 0501 Table VIII- I9PID4v.2 C' Terminal-Al -9-mers Each peptide is a portion of SEQ ID NO: 5; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight. -Posi Subsequence Iscorel i~1LRTTIE 0 =12j 7il DTLI 9J [III VHTRPDS1[.010 1 _j RPT-1DSRST 0.003 [1j STCE F-4-11VHTRPTS 00i1 o[Eq ooi0- FEL_7TDSRT FoEdo=6 Table Vill 109PI1D4v.2- N' terminal A1-9-mers TalVI 1 09131 D4v.2- NtrIna AI-9-mers Each peptide Is a portion of SEQ ID Nor. 5; each start position Is specified, the length of peplide is 9 amino acids, and I the end position for each peptide Is the start position plus IF1 0 L-1 F-0 0 G I2II~TVSVG 0 .003 2[39 GLTVqt]L90T1 IQQVTSP =0.003 Tabl VII- OPI LFk-er ID O:FgI 7; eahst oos poiE o ach peptide is th e start position plus eight uence S 7o re_1 11061 NSDPESTFI 78 11=1 [l~ jqF ~t7 NQELI] I ~9Ki 0 900 go,[ 0.900 F11 32ILKS NC T qEC q]E.50 7Q 288SDGVG 15411 SLDHSSSSQ IF 0500 Each peptide is a portion of SEQ ID NO: 5; each start position Is specified, the length of peptide is 9 amino adids, and the end position for each peptide is the start position plus eight [POS bsqence =[Swe R=QWVLl =0450j ET TSVPGM DL LJ.5 =!VrPGMDLJ[015 I EPI V PGMDLLS FI.:959.

[.TII Fv:L:.LQQV f0.05L7 FI D11 LQQvTSY I[ 00 PFT VIQIQL!I 0.010 Fi- I FQCL 01 q 010 00 00 Table VIII 109PI D40.

Al .9-mers Each peptide Is a portion of SEQ 11) NO: 7; each start position is specified, the length of peptide Is 9 amino adds, and the end position for each peptide Is the star posiiion plus eight 1l3-71 TQCLIYGH 4 .201 LEGQESS~ 0.I2[:251 [TI]E G NS 0 225 P-MJi SPPVAI [.-150' 170-N HS, PsQA s o U18211 Y SPP Q 0. 0150j .S ffSQESSDG9j 0.135 III1 QGS VATsQF 0.125.

1 36 SPPVILJ 0. 125 F25861IPEYDI 0. 12 5 69] DGLT .2 Fl198 .TTII.C 2=5..9? 258 DQVIAL-H RJ012 1333 RGSMEHf_025 TL! CTPMKS IL_.10 f3-16 KVPTF .100 13q7 I RLHPSDDSI 0?900 1241VPEA 000 J 'i4-1 KVAGKSQR .100-- PS§DO-SIKVI. DL 0.075- .iF6JLTST§HPLFk. oQ6oi FP-ls:TmEw- !L 29-5 yj0.7 25LSPLQII007 Table -VI- 19P1D4v.31 Al-9-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, t length of peptide Is 9 amino acids, and the end position for each peplide is the start posi tion plus eight h22]LVQATALHH IL.0~ 17- STSHGL5PL 1 0.050 [1j[LTH: PlPLQ~ 0050 I _j [07~ ffRLPLGYflF 0.50 2!~ITPHHSPRVI0.0 246 LIAA6JSH ILE9700iOF L162E QQASALCHE L E6 6i D~LL -YE-J1 o-osoI [1Ls][kP PIA II.0~50 I [0]L YYRSCT 0o4 D88 FEEYF=RT 9.045_ 157 FHSSSSQAqJ =0030 255][SSPLPQVIA 1_ 0.030 169 E OSSQAQA SA 0. =3 W34[M-SERLHPSD 0.027 1271ASQFYTMSJ[ 0.025 149 2 J005 lii TPKEV 0.2 L1O5ILGNDPESTfl 0.025 F9-51TP=SNRTEG.2 275CH-, SPPPIqV ij .0?5 jFIJJ17 'LHS 11 0.025 17j MES1 1J 0.025 215 ALHSPPLV 0.2 167 C9SPP L .2 IjTable Vill 109PI D4v.3 A1-9-ms Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the ~I ubse ~nce LSqoreI I 190[~CHSy=.=020 kE49 LHSPSJ 0.0201 J SjQWQ .020 1-9-21 LCSPT L 0.020 [2&jj~qHSPPP-1ji 0.020] 661 K GLGDH DA Gj0.2 I F47DAcWMPASLJ020 Table 'All -IO9PID4v.4 A1-9-mers_ Each peptide is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide is 9 amino acids, and te end position for each peptide is the start position plus eight.

os {Su bsequen HPPQSQR 250 ULL tIHQPaSQR DDt ]F 001 PQPQSQRRV 00=0 ji Table IX 1O9PID4v.4 1-1 Ilers Each peptide is a portion of SEQ ID NO: 9; each start position Is specified, the length of peptide is 10 amino acids, and the end Iposition for each peptideIs the start position plus nine.

=PsL bsLec jF-3-1 WIHP PQSQR ,-Oo F7 _f_9_k Ry F FO050 00 I Table IX 109P1 D4v.4 1Table XI 109PI D4v.4 L Al-1O-mers J A0201-10-mers CK1Each peptide is a portion of SEQI Each peptide is a portion of SEQ ID NO: 9each start positionIsl ID NO: 9; each start positionis specified, the length of peptide is specified, the length of peptide is aino acids, and the end 10 amino acids, and the end position for each pepiei h position for each peptide Is the trtpsiinplus nine. jI satoion lus nine.- [jF~P qSQfl j lSor F sj qRRVT IFoGro F.I H QSQRR FTFHLI 1oa w 1( IH Pt SQRR Lo.Km- ELLIwh~o~ 0.002 DD7i][ HPQnQ Ir~o]Table Xii 1O9PID4v 4 F6-j PQPQsQRRVT J(-9mei =8 F _PQSQRVTFHJL900I0 Each peptide is a portion of SEQ ID NO, 9; each start position is specified, the length of peptIde is 9 Tabl X-19P1Dv.4amino acids, and the end position A020 -9-nersfor each peptide Is the start Eac petd-saprino E posftoiplus eight.

specified, the length of peptide is9. s L §!9PseqRR1[-F-GI amino acids, and the end [ositio 1HQQQ for each peptide Is the start =3 F9__ p~gan plus eight. j7[PSRVFJ FPo0s I ub uenc e I[h[IHQQS3 =2iii WIHQQSQ 7-E- [010- 7QpQsQRRVT I PI 8I QSRTF 5[ 1 Lj II[ IW1HPQPQS JE00 LEJ[_QPSQRRVT_1L00=4 IEL R±J-o000 [jj1 PQsQRRVTrF 1F55 6.666___ F2_:1 1]L Table XII]I- 1O9PID4v.4 __PP SQ f Each peptideIs a portion of SEQ specified, the length of peplide is Table XI- 109P1 D4v.4 10 amino acids, and the end A0201-10-niers posito for each peptide Is the Each peptide is a portion of SEQ* start position plus nine.

ID NO: 9; each start position is j=Squen e specified, the length of peptIde is i~ SRE 0[70o amino acids, and the end position for each peptide Is the I F7 Lqiqvr1 1(0.020 sta!rt position plus nine. I VTFL!013 fsIF Subsequence--F- [(-TeJ r71 I~P S Ilfi957 LT] Ei~hPQP§J(0.06j I EIWqhPQPQ FTI[ HP~qSQRR~ 0.0031PgqqsRRVj[00 F-8j PQsQrRvTFH 0.002 tw IMHpPQSQ]F oooI-0

'I

Table XVI 109PI DUv.

A24-9-niers Each peptide Is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide Is 9 amino adids, and the end position for each peptide Is the start position plus eight___ 1 IWIHPQQ Fo-1-o( [AhHQPQSQRR [02 =6IPLP~s§@ yVI ko-1 00- Tal eV XXI9P-Dv. 109P1D4v.4 A24-9-mers JTable XIX 109131 D4v.4 851-0-mers Each peptide is aporion of SEQ 137-1 0-niers i Each peptide is a portion of SEQ ID NO: 9; each start position is Each peptide Is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide Is 9 ID NO: 9; each start position Is specified, the length of peptide Is amino acids, and the end position specified, the length of peptide Is 9 amino acids, and the end for each peptide is the start 10 amino adds, and the end position for each peptide Is the posiion lusei~h __position for each peptide is the start position plus eight [FP ]S ubeuec I Score start positio pus nine.JPo [ueqejcrI 015 Subequnc L Subsequence_ c-i s IPQPQQRRVi 0.015j F-L o II QSbrTs H ]4.000]r tWhQPQ .0 rarT1 I ETII f pso _UTj i 1 [3 ]IHQQSR~fJ IFT-1 D=-qRVF1a00 LIj WIHQPQS Ej0.1o F-5--]FCi~-q P .1 00 E=WhPPQ E~I LIi[E i1o(ooi ITable XVII 10913134.4 1 IT] IH PSR05] F2 IW IH Q00011 I A24-10-niers L~L QQR RVT 10.015IP QQR .01 Each peptide is a portion of SEQ jK I El -QS Q r R V TFH J 0 oD N0r 9; each start position is 2[IH.iQQ 11 U001 1 Table VIII-IO9PID1v.5 secified, the length of peptide is _M 0.001_ amrdno adids, and the end [=4LHQ p SQ RR j~~~TI IAI-9-mers positon for each peptide is the -Each peptide is a portion of SEQ str Tablen nX IOPD~. D DNO: 11; each start position is Su uenc Tal___ 19I specified, the length of peptide Is L=9 L.9 RrvFHL [8A99J0 Each peplide is a portion of SEQ position for each peptide is the 1 L IEwmhPQPQKSF 1010j for each peptide Is the start [3ISVHTRPSQR] 0=00O III L-Wj L i pq~g Lop-0i s pston plus eight 3ZLQRVT IIZPQPQsQRRV 0. 015 jube c SC;[T VSVHTRPSQ 6 .3 OLDWI~PQQ 11.0121 _7.1 gPQSQRRV !fi Fj[ TRPQR 002 FT]1 IHOQSR I0002 A [71112n H~PSR .200j' L i PVSVHTRPS001 LLE 1[IQ T oj~I L71..PQSqRRVTF 10"100] L ILTR9.R ].601 1 [8 J[QSQRRVTFH 10.0501 [7K PSq3RVT.jooj Tale XVI 11- 10PID4v.4 5[ PQPQSQRRV 0F.020 IL QRVF I[J L B7-9-mners j 1 IWIHPQPQS 0.0101 Each peptide Is a portion of SEQ 1 I iiLVHQ QI =0.0101 1 Table IX-IO9PID4v.5 ID NO: 9; each start position is IHQQQ r specified, die length of peptide I 9 ]KFl LHQQQ..01 lI0m amino acids, and the end position Each peptide is a portion of SEQ for each peptide Is the start jTable XXI IO09P1ID4v.4 ID NO: 11; each start position is po ionpIy .hl4 II ____o-10-niers Ispecified, the length of peptide Is ,Foi['-be -iB50 10 amino acids, and the end pep-tde is a'portion of SEQ position for each peptide is the 7JL99~g~r ~ID NO: 9; each start position is start posLion plus nine.

If 2.7 specified, the length of p~ptide Is; 1117PPQSR 0.020J i position for each peptide Is the g L o~oi 01 L start position plus eight I pQ f0.01 ;[71i IHPqQsQ 1L0.olols~q~no JsoT IHTRPsQRRVF 7X1[2oRri [_6jisqece~cr~ L. T I 0025 11f WHPQPQS 7~~o IL TR~RVF0aoio 1311 I PDPO5 oR 1-1-QQ0V F L 01I3 ~i I 00 3 00 00 Table IX-109PiD4.5 Al-I 0-mers Each peptide Is a portion of SEQ ID NOr 11; each start position Is specified, the length of peptide is amino adds, and the end position for each peptide is the _.start osniplus nine.

i[ I bLSjequence Score LX1 SQrRvrF~III 9.0 L fIPS?~QR SVHTRpSQRR .0 Table X-1O9PlD4.5 A0201-9-mers _1 Each peptide is a portion of SEQ ID NO: ii; each start position Is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start I _p plus eight SVrRPS(R 0.001 D I RPSQRRV I00 PSQRRVFH f 6 I i1L QRRVioo 1 PVSVHTRPS 0.000 Sii] IP E0SQRR i001 Table XJ-lO9PID4v.5 A0201-10-mers Each peptide is a portion of SEQ ID NO: 11; each start positlon Is specified, the length of peptide is amino acids, and the end position for each peptide Is the start position plus nine.

9Pos Subsequence s-ore 9 QR FHL i0.018 ;[Tii VHTpSRV 10.016 18 LILP _WVLF F 0.006] 1!177l SVTrPSQRR IF O.ool1 =F1_ VPVSvHTRPS j[0 00] I 3 1 VSVHIRPSQR 3[60] 1PVSVhTRPSQ 0.000 Table XI-1O9P1D4v.5 A0201-I 0-mers Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the startpsition plus nine.

[PosI SubsequenJ[Score Li6 Hiri ji '96-61 Table XII109P1D4v.5 A3-9mers Each peptide Is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight PosI Subsuence 7 Score I13 SVHTRPSQR I0A00I Lii RPSQRRVF 0920f SHTRPSQRRV [5Oi SpsoRRvrFH 6.626 =8 =SVTR~a10.000 1iPSVTP 0.000 ILELTf9RRVT 0.000 Table XIV-109P1D4v.5 A11O1-9-mers- j Each peptide Is a portion of SEQ ID N0. 11; each start position is speified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight FPossjsbeu _TL R06 [1F-PSQR .030 877 RPSQrRrI 9I0.002 j[7 TRqRRVTF]O.002~ F'F11PSQRrvTFHL 110.0011 61 L HTRPsQRRV J 0.001 j.S._!v!HTR S 1 0.0001 ID 7L\TIVIvHp9R ELT][ Me RV .00 Table XV-109P1 Al101-10-mers Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amIno acids, and the end position for each peptide is the start position plus nine.

[Pos i iSus e[ score SVHTrPSQRR 0.400 3 7F_0_SR 00076 MLAPAQrRVTFH..

JU.OQ]

M PSQRFHLJ[ 0.000 02( VPVSVHTR 0.00] L TRpSQRRV =0.000 Table XVI-1O9P1D4v.5 ~A24-9.-me___ers Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus eight- F o i o Susequncel Scre 7 RPSQRRVTF 14.000

ED

7 HTRPSMRRVJ[0 12 WLI1 RPPYLJ 1 0.015 L ilsv.tPsQ iLo~i_ L7I]LPvsvHTRPSI~sI 0.010 WL..l o1~PlRJ W I PSQRVWH JL002 EKLl VHTRPSQRRJI01 00 00 Table XVII-IO9PI D4v.5 1 A24-10-mers Each peptide Is a portion of SEQ ID NO: 11; each start position Is specified, the length of peptide is amino acids, and the end position far each peptide is the start position plus nine.

-t1 ESubse q Iencrel II RPSQrRVTF 0[? 3 ]Ly TrPSQR 0.015 [ILv vspsq-1 iool L2 PVSVhTRPSQ 110 ooij L Table XVIII-1O9PI D4v.5 137-9-mers Each peptide is a portion of SEQ ID NO: 11; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus eight.

Ijojj Subsequence JIScore] 1 HTRPSQRRV I2.j 7 RSQRVIF 0. 600 WVHTRPSQR~j 0.050 E2 Ln VHTRPSQ IE 15J 16] TRPSQRRVT 0.015 L i[_PvSRPSl0.010 4 VHTRPSQRR j 0.002J R~H .00, Table XIX-109P1D4v.5 B7-10-mers Each peptide is a portion of SEQ ID Na. 11; each start position Is specified, the length of peptide Is 10 amino acids, and the end position for each peptide Is the start position plus nine.

_L.LI.J HT~RPsQRRVT 11.500! 1 9 JPSQRNrFHL -104 VPVSvHTRPS J[0Oi VHT110.02 VSVHRP .010 I =2 LP\SVI Table XIX-1 09P1 D4v.5 B7-1 0-mars Each peptide is a portion of SEQ ID NO: 11; each start position Is specified, the length of peptide is 10 amino adds, and the end position for each peptide Is the start position plus nine.

[Pos DI -bsequence LI TRSqRRVTF13 Table XX-109P1 D4v.5 3501-9-mers Each peptide is a portion of SEQ ID NOr 11; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start p on plus eight Pos Subsequence Score, 77 RPSQRRVTF 5 jjt TR YJRV .0 SVSVIHTRPSQ 22050 [7[PVSVHTRPS 0.010 ;8][PSQRRVrF 0jl0I E -=1LY QRPSQRR 0.001 Table XXI-1 09PI D4v.5 83501-10-mers Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino adds, and the end position for ea peptide is the start position plus nine.

=Po ubsequencei rScore W VPVSVHTRPS 2.000 W PSQRrVTFHL LP.5P0 RPSQRVTH 0.400 17 Rr9R~RV 10.3001 0.050 i~5JL~i7pLJYV10ff21 010 IL PVSvhTRPSQ 0.001j Table VIII-109P1D4v.6 C' terminal-Ai-9-ers Each peptide is a portion of SEQ ID NO: 11; each start position is specifed, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus eight.

Jjuos=L ScenT] core HTRPTDSRT [Lp.025 FL 1i_ SVHTRPTDS 0.010 F. T L\ SffRff 0.003 [f PVSVTRPT ][0.01 S I VHTRPTDSRIR -67 Table IX-i 09PI D4v.6 CUerminal-A-lO-mers Each peptide is a portion of SEQ ID NO: 13; each start position Is specified, the length of peptide is amino acids, and the end position for each peptide is the start position plus I nine.

Vf- EL -Subsequence II jrej 4 i SVHTrPTDSR 0.100 [I7LYVYVPIP.DSI~:01 IS~RPD =1 VPVSvHTRPT 0.003 271 PVSVwn TD =0.000 VHTRpTDSRT I[ O]00 Table X-109P1D4v.6 C' terminal-A02011-9-mars Each peptide is a portion of SEQ ID NO: 13; each start position Is specified, the length of peptide is 9 amino adds, and the end position for each peptide Is the start position plus eight -Po Ii subs]que] Score =3 SVHTRPTDS 0.007 =i PVSVHTRPT 0.003 j 1 HTRPTDSRT D0O] I 2. VSVIITRPTD 0.000 [1.4 i VHTRPTDSR II 0.000p 1 Table XI-1O9PID4v.6 UC terminal-A0201-110-mers Each peptide Is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide Is 10 amino adds, and the end position for each peptide Is the start position pius nine.

00 00 1 ubsequence IFScare T][1 VPVSvRPT IEEE17 2 1[ VSVHtRPTD 16.667 Table XII-109P1D4v.6 C' terrnlnal-A3-9-mers Each peptide is a portion of SEQ1 ID NO: 13; each start position Is specified, the length of peptide Is 9 amino adds, and the end position for each peptide Is the start -position plus eight__ FP- -1

TPTSR

74 VHRPDS =3Fh svl-nRrnlS]F( .004 72]v-- m[~oo [7I7JLv~sffx-PT ][o-oLK Table XIV-1O91PID=v.6 Cterminal-All 01-9-mers Each peptide is a portion f -SEQ ID NO: 13; each start position Is specified, the length of peptide is 9 amino adids, and the end position for each peptide is the startj __pqsItionylus eight.__ Table XV-109P1D4v.6 L CtermInal-Al 101-110-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino adids, and the end position for each peptide is the start position plus I_ nine.

[4 SVHrPTSR 0.[ 1FfiY-LvfHTR7T oo C' termlnal-A24-9-mers Each peptide is a portion of SEQ I D NO: 13; each start positioni I specified, the length of peptide Is 9, amino acids, and the end position for each peptide is the start position plus eight DP j]i S bsequenceIL =12 F-3ISVTRTD 0.100 21 FVSVHTRPTD 0.015] Table XVII-1 09P1 D4v.6I C' terminal-A24-10-mersj Each peptide is a portion of SEQ1 ID NO: 13; each start position is specified, the length of peptide is amino acids, and the end position for each peptide Is the satpsto plus nine.

jj__utec e! Scorel 1=1I] VSVHR~Tl.-s 11.150] Table XVII-1 09P1 04v.6 C' terminal-A24-1 0-mers SEach peptide, Is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino adids, and the end position for each peptide Is the start positionypu ie =DoS~lusqe ]i crel 11271 S[VH TrPTDSR j 0.01 0.001 Tale X~I1 09P1 D4v.6 C'tria-B7-9-mers Each peptide Is a portion of SEQ ID NO: 13; each start position Is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start pjositon pus eight.

[PqJ Suseuence liscorel =_5211 SH TRPTDS I-i1001O Tabe YlX-19PDa6I C, terminal-87-i 0-iers] Each peptide Is a portion of SEQ ID NO: 13; each start position Is specified, the length of peptide is 10 amlno adids, and the end position for each peptide Is the I start position plus nine.

EP- F[Suseuece Scre] 1 VPVSvHTRPT zo] =4 l vrnR j =0.075 3J PVhTRPT o-.00 dTable XX-1 09P1 D4v.6 CI tri laM319-ner.. Each peptide is aportion of SEQ IDN:1;each start position Is specified, the length of peptide is 9 [amnino adds, and thue end position for each peptide Is the start postio plu eight- Table XIV-IO9P1!D4v.6 1 C'terminal-All1 01-9-mars Each peptide is a portion of SEQi ID NO: 13; each start position is specified, the length of peptide Is 91 amino acids, and the end position for each peptidelIs the start poiinplus eight__ IL4 j VHTRPTSI .f0.0 3 IfSVHRPTDS 10.002] I-ITRPTDsRT F65601] 00 00 PFi 11Subsequence Score W HTRPTDSRT 0.300 WISVKTRPTDJF01J0 2 VSVHTRPTD J.as M L tv-§ y 0.0101 LWLY!LRTDR 00II oof Table XXI-109P1D4v.6 C' terminal-63501-10-mers Each peptide is a portion of SEQ ID NO: 13; each start position Is specified, the length of peptide is amino acids, and the end position for each peptide is the start position plus nine. Fos Subsequence jScore muVPVSVHTRPTJI00J VSVHIRPTDS l.soo WLSVA!IPTDSR j0o0o] VHTRpTDSRT IfFo] V PS~TRPTT 11]0 Table VIII-lO9PID4v.6 N' terminal-Al-9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight fPos aSuceqe L Score W NSDISSWR 15.000 23 L.U!!GTYIF 10.200 14 RVN1TNCHK 0.200 LL!LsswRv\l 0 .030 16 INTNCHKCL I[P51 17ZLTVGFNSDI 0.025 L2 11 HKCLLSGTY 10.025 17JITNCHKCLUjI.025j LJO[SSWRVNU 0.015 VGFNSDISS 0.0131 8 INCHKCLLS 0.013 {T ITGFNSDIS j.010j .L J KCLLSGTYl 0qi10 19 ii NCHKCLLSG 0.005 SFNSDISSW I T003 VNITNHKC _0.003 Table VIII-109P1D4v.6 N' terminal-Ai-9-mers Each peptide is a portion of SEQ I) NO: 13; each start position Is specfied, the length of peptide is 9 amino adds, and the end postion for each peptide is the start _.position plus eIght I =O sI reubsequence IIaI F SDISSVVRV 10.001 F 11 SvvRVNTN [06001 [12 VV-~~i~RrNFN~ J[0.ooi L GFNSDSSV~ o 0.001 L713-J VRVNrTNCH JL(P Lo 20 CHKCLLSGT 0.000 Table IX-1 09P1 D4v.6 N' terminal-Al-i 0-mners Each peptide Is a portion of SEQ! ID NO: 13; each start posItion is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine.

Pos Pi SubsequencelScore [6][NSDlsSMV 22i[1LCsGTYF gq99i D7 1NChKCLLS Jj.I- 2 1NSDISWR K0--%Ih I LTYG-- 1sR! s- Table X-1 09P1 D4v.6 N' terminal-A0201-9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus eight.

Pos L Dsu iiie l c Di KCLLSGTYI 1 -M iI L MVGFNSDI 0.936 16 NITCHFO 297 17 CHKCLL 297 LriLISSWRV I23 IL.SGTYIF I0113 I J~IL _Y.YL 011 =3 I ISSV 10.11 79JISSWRVNTI~ LYfj12 .056JMM cHK E66 51 SVVt SRVNTTN 0.0 f 2 TVGFNSDIS 0.001 LiLI NCHKCLSG 0.001 =9 1 NCHKCLLS 1 CHKCLLSGT 0.000 8 IV E000 13 VRVN1TNCH 21 HKCLLSGTY Table XI-1 09P1 D4v.6 N' terminal-A0201-10n-mers Each peptide is a portion of SEQ ID NO: 13; each start position Is specified, the length of peptide is amino acids, and the end position for each peptide is the start position plus' nine. Irw~l'i sbsence 1l LScor2i 1F. 23 1151.6481l~!!.( B lLJLYVG F!0LvJI 6.568 1 i RVNTtNCHKC JIP.435i jil SVVRvN1TNC 0.435 _S I yyRV 0.4181 L1iJLVYKTN K0. 10.01 4 R RVNTCHKC lml1 20 F CHKCISGTY o7o3I [Vl'NTnHKC 0.3! ?.LiIdNCHKcLsG.0o1 LIWRnTNCH Q t ii il(Svv~NrNCllo0.001j F_7_ SDISsWRVN 1 0.001 GFNSdISSwyJ 0.001

I

00 00 Table XI-109P1D4v.6 N' terminal-A0201-1 0-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide Is 10 amino acids, and the end position for each peptide Is the start position plus nine.

Pos Subsequence' re 16 i NTTNcHKCL L .297, VNTTnCHKCL 3 9 LjDSVVRVNTT L[O7 jNCHcCL l sGT~ L P -is vnI f =-on [7J GFNSdISSW I[ 0.020 1 [Lii TVGFnSOISS I 1(07J0 21 [jCUSG1Yl ]j 0.003 I 22 KCLLsGT'IFi 0.0031 TC~ltCLLSGjj 0.001 [17 11 rrNhKCLHS!o.011 [12 iWRVnTCH 0.001 LI~I71LSDISSWR O19. lI 1 MTVGINSDISf 0.000 12I KDE00:01 3 LIV0TNCHK 11 0.00 Table XII-10911)4v.6 N' teinal-A-9mersf Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start L sit onypus eight Pos Subsequence Score F23-1 I TY F 9.0- 0' RVNTTNCHK I2.000 i 1 j[ MVGFNSDI 0.203 F 17 11.TTNCHKCLL 0.030 1 t2tCU~SGmYI 0.027 =2L7. NSDISSWR flo020I f2W RVNTNrC 0.0201 16 !I NTTNCHKCL F 0.015 if SMWh Ii 0.005] STVGFNSDIS F05W4 ISSWRVNTT .0 Table X11-1O9PID4v.6 N' terminal-A3-9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight.

I

Pos L bseence1L~scr2 [LLJIjLY DLO90l W SDISSWRV 0.001 1 L 1 F!SPlSS. j[ 0.001 I =4L 11 ISSWRVNT ]L F. 1 F IIL1NKCLLSG o.0j 1 FNSDISSW l.001 L 15i.yfVNTTNHKC 1f 0.000 13 YRVNT~h9!iJI0.OOJ1 L8 DISSVRVN 0.000] 18 PCqHKCLLS 0.000 10[ CHKCLLSGT J100P Table XIII-1 09P1 D4v.6 N' terminai-A3-10-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is amino acids, and the end position for each peptide is the startposition plus nine.

I Subsequence jScre L 1. _LCsLL yIFA o10.600 o L KCLLsGTYIF .279J 13 VRVNtTNCHK 1I0_30i [1W INTcHKLL 030 ii 1SVRVNTRNC 0.030 SRVNTCHKC .020 12 WRVnTTCH 0.020 EI~LF i!1 R0 00 2:_i i _TVGFnSDISS 0.008 S_ 1 !I MVGfNSDIS 0.005 1. 8 [1ISVVRVNT 110.

17 -L P~~i OP~ F F7_ J_11 L.TTNChKCLLS.0.0 I E LNSIIIsq\ V 3 11 VGFNsDISSV~ 110.002 1 71] ISSVvRVNT 0.002 19 1 _NCHKcLLSGT 0.002 SCHKCLSGTY .001 Table XIII-19P1D4v.6 N' terminat-A3-10-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine.

JPos[ usqec 4L i GFNSdSSW JLO.0011 F[15][ VNTCHKCL 0.0011 l7 7[_HKCUSGTYI j pq EE] 10-NT~ 0000 18 TNCHkCLLSG SDISsVRVN 0.000 Table XIV-109P1 D4v.6 N' terminal-A1IO1-9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is I specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight Po Score L I4JLRY! U CHK IL6.000 ;LiI] MWGFNSDI jL0.015 I IFiL1_ 1j fiL, Eo E2. I [1711 TTNCHKCLL 10.010 I 2=2 KCLLGqTI j=Nl r4 GFNSDISSV 0.006 16 -1 NTTNCHKCL I 0.005 [6 NSDISSWR [0.04 11 [SRVNJ ]L00i 12 [1 VNTNC 0.002 2 LTRNSDIS LF002 19 NCHKCLLSG ]FO00J 5 L LSD_ff= C 0 0=0 SDISSVVRV ]0.000 1311. VRVNTTNCH .1 0.000 [5 IKOSG1Y I0.000 1 [1~7JI..vGN~ls I [o.oooj .LIIJI STNCHKCLSIIO.00i 9 SSWRVNT 0.000 20 I CHKCLLSGT F 0.o 000 lii DISSVVRVN ]I 0.000 00 00 Table XV-109P1D4v.6 N' terminal-Al 101-10-mere Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino adds, and the end position forl each peptide Is the start position plus nine.

LP]Ls ubsequence Score E-3 VRVNTC HK .030 1271 WRVnTCH- 0.020 IF'1 KCLLsGTYIF 0.:1 EFLJ1CL-LsgDYFA (.F1: DE,,7_NTTNcHKCqL 0.010 i FNSDiSSWR ]a008 [Th7[ RVNTNCHKC iL.I0i F 71NSdlSSW [0.006 L2 1VGFnSDISS {05j _7 1 SWRVN -TNC 10.003 7 TrN >N~KCLS '10.002 LI MTGfNSDIS 0.002 L- i VGFNsDISSV j[959] =9 NCHKcLLSGT J o0.0 WI NSDsSWRV j 0.000 KCILSG Y 1[9- O 115 LV1TnCHKCL [0.000 T DISSVVRVT j 0.000 [T 1[ TNCHkCLLSG [i O9' io I SSWrVNTTNW WI_ ISSVVRVNT 0.000 SOISsWRVN fj0] Table XVk-O9PID4v.6 N' termnal-A24-9-mers Each peptide Is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus I eight PobsequenceII Score i 17 1qNttN q! I 6.000 1 [i31 CLSGflIF j3.000 F2KCLLSGTYI 3.000 MGI FNSD I L Table XVI-1 09P1 D4v.6 N' terninal-A24-9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus eight FI4I GFNSDISSV L L.m5J 'iLF-N§vI§§w AT16 8i ISsRT 0.140 10ISVN1TNCK 0.110 ffV[SDISJI O.10 18 ITNCHCLL 0.100 ]GFh[SQSS! o.io 2 1 F R)N mcLIo.o F4-I RvN T- EomL [l iSDISSWRV 0.015 ~TJ HKCLSG1Y001 20 ICHKCSGTJ.12 I1 NSDISSR 0.01 FINRI Ot "7' 19 0.0 10 Table XVII-1 09P1 D4v.6 N' terminal-A24-10-ers Each peptide is a portion of SEQ ID NO: 13; each start position is I specified, the length of peptide is amino adds, and the end position for each peptide Is the start position plus nine.

L j i; e n e11scoreI I KCLLsGIF 6. 0 VNTKL :f4:O L ;L FNdviY I. jOS*j L i RVDwn-NC K 6.336' 21 i L a'rvSrNJ 01oI fiilj TNChKCLLS :0.~W 11 SVRVNTNC 10.10 Table XVII-i09P1D4v.6 N' terminal-A2410-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amIno acids, and the end position for each peptide Is the s position plus nine.

SPos]I Subsequence].Sore LF.I -DvISSvVRVNT 55O.140 ISSVvRVNTT j0.120 [7isl .NCHKLGT 110.120 L .I 1VGF =1009 Lull__ NSDsSWRV .1 LI[_VGFNsD ISSV J0.100 LSDIsvRVN 110.0211 2= 0 CHKLSGTY l0.012 FNSDISSWR Fi.012 [uh WRVnTRmCH 1.0121 [18 NTNCHkCLLSG IFaoi I3_1 jIR~Th9HiI0002I r Table XVIII-109P1D4v.6 N' terminal-87-9-mersJ Each peptide is a portion of SEQ1 ID NO: 13; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position plus eight D5 sl SuI qec I[ScoreJ 12 1 WRVN=6NC-000 (JFILNUNCI-CL] 4.0001 L7 IL± 2 1 G: 0.400 =L _FNSDISSWJ0 L 1ISSWR v T 1II I101 SSWRVNTT IO109j IEi 1. imP IWNTN- Eno *0 _.15 I 1 L[RVNTTNH jg Plsq TVG_ N N 0.10 I!al DISSVVRVN__j0.?O0 4 GFNSDISSV .020 F TCHKCLLS j=.020 I7 ISDISSWRV .020 00 00 Table XIX-1 09131 D4v.6 N' terminal-B37-1O-mers Eachi peptide Is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is amino acids, and the end position for each peptide Is the start.positlon plus nine.- Pos [I7§!LuenceJ[cr J1 VRVNtTNCHK- .0 rf Table XX-1O9PID4v.6 L-N' termlnal-B33501-9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of pepfide is 9 amino acids, and the end position for each peptide is the start ___position plus eight.___ =616j NUTNCHKCL F.000 IL]L=1KOLL00o Fi97 IL 5P~'~[.00 F JI~ I 1 2] TVGFNSDI-S] 0.100~ 1=8 L MqtKCLS 0.100

LYNTTNHKI~

Li]IVGFNSDISJ 0.-100] L iKCLLSGTLQO =4IFNsIssv [_lo NSDISSWyR (0.015; STable XXI-1O9P1D4v.6 mna-351- 0 r J1 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide Is 10 amino acids, and the end position for each peptide is the s -tart position pus nine. Ijp~ J Sbsequence ',Lcoej L~i~ 19CL~GTY F 2.0001 116 N 1T cHKCL{70 15 VNTTnCHKCL '11IR0 D E2 1 1. PCHKsGTy j.6 I CE §V~n1-TN 171111 VGFsDSV [0.300 L I4 J0.200HK I 19II NH~cLSGT] 0.100 =2~j TV]SDS Eoio =81 ILRssv VvNT oEioo[ L=1 _MVGfNSDIS IFpOOI0 11723 IT CLL~g1YIFA 10.1001 ~L2L IL~rN~hK L L ±I §WljvNTTNc Ej. 49o 6 21 HKCUGT] 0.040) '[_Mn~tCH L FN SSW 2 0 18 !ITNCHkCLLSG 001 ;LDsWV iio 13 [1 VRVNtTNCHK IF0.0011I Tabele II-1O9PID40.

N'ternmalkAi -9-mers Each peptide Is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide Is the start _positkon plus eight L sI Sbsequence [1[Scorej .14 SLSPLLV [0.5w_ I 1 [LSPLLLVSVy 0.030[ 11 SSSSLSPLL 0.030 PLLLVSR JO0020 [.18 1 =LVVR .920 00 00 I Table AI11-1O09P1 D4v.7 N' terminal-Al -9-mers Each peptide is a portion of SEQ ID NOr. 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position puseighL I~yjL~ jFJj ss-1j F0.0151 F 1-9 1L y-t-1Lo 0.010 F-7 I FL0 sSs] 001 L7111 vGftiSsJ =0.0031 '[Z16 0.003] EI EL .L!I ss...1 ii 1 MFRVGFUI i=.o.0I9 L Table D- 09P1 D4v.7 N' terminai-A-10-mersj Each peptide is a portion of SEQ ID NO: 15; each start position Is Ispecified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine. 1=2l sLs- wv lEq..~ 111 SSS sSPLLI 0. 0=7 16i SPUVSW I .05 EI9- ]I LVSVRVNT ]a.07:20, 1fF4 RVGIISSS 5 o-919 RVGIISSS =~0.010 W I IIS~SSLP01 0.

Table IX-109P1D4v.7 Each peptide is a portion of SEQ ID NO: 15; each start position Is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus GFRVIiSS I0.001 S Table X-1O9PID4v.7 N'termlnai-A0201-9-mers; Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 ano acids, and the end position fo ahpeptide Is the start ___position pius eight.

Poe T if Ebsequene fl~]L Rs IFi L~iLL ySx 3 ILL. 8.J [10\EN' IL s C1L.1i] L2LIEC VW N1TJ .190 L1i01L SSSLP~ i 0.3 1 F1-2-1[ SSLSP LLL ][0.139 I 14J[SPLLL--V:LO070I 19i ELLVSRY!R I =0.024 I LIS§SSi LS O 1 WL li 0.015 I 7ly-GFLI§sSS 7FF001 [TTI MFRvGFLI 09.661 I I J5 i VS [7F§O Ta[le 1 09P1IISSvf.7 IF NtrinalF3GuIAO j~ Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide Is the stitositio plus nine.

L I_ SLSPILLVSV- 1I599701 [i~i~li F~sSSSL 198.2767 I- 17 IP LLLvII3.022 2=0 1_V vRVN1T =I 2550] L1=L yLvw JToJ28I 11 I SSSISPL~jI .139 F-1]8[ LvVRN o-0088 121][ UI§SsSSLSP IIJ-0 j 1111SL~SLLPL O.00-2] VS2 rVNT[ J[001 =[_FLfSqs-sS][ OOOOJ 1=6]1_SPuWVSVVR][0.000] LT1LFRVG -!iSS FOL0] [i[MFRVgFUJIS (0.000 1 Table YI -109P1 D4v.7 N' terminal-A3-9-mners Each peptide is a portion of SEQ ID NO: 15; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start P okcsItlon plus eight. [Ps[Subseuence Scre] f 17~[ LLLVSVVR 1.900] S77 PLSPLLVS108] FLISS L-0-LLVSWRVNT.. KO [RVGFLIISS 0.0 12 1 ][SPLLLVSW I 0.009 13i i[ ]SSLL 0. 00 7 00 00 Table XII-1-09P1 D4v.7 N' terminal-A3-9-mers Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptlde Is 9 amino acids, and the end position for each peplide is the start position plus eight L osiI Subseq jeII ]x Lb =0 SSSSSLSPL FC L8_j1L_ lySySSJL~Jo CCLMF~~!~ 0.004 11 LSPLL F .90 3] L ILFR FIIS 0.001 FT7I VGFLIISSSJ 0.00-0 JL iGFUIISSS 1=.9-6 Table XIII-19PID4v.7 N' terminal-A3-10-mers Each peptide Is a portion of SEQ ID1 NO: 15; each start posItion is specified, the length of peptide is 10 amino acids, and the end position for ch peptide Is the start position plus nine.

L Lriub ce Score FLIIsSSSSL f 0.9 14 SLSPILLVSV jj 0.450J Ef 9 flVRVT0.2 E17 APW S.0 9 =1 j spw sw j[of 0 iLVSVVRVNTT ]0O3 is II L i 1 0.013 I3 itRVGIISSS ][j0j9 L0SsSSSLS ]0.06 ii SSISPL .006 F[1!2 LSSSLsPLLLV J[ 0.05 ISsLSP E 00I IL0i SSSSsLSPLL {i U03 ii LSPULVSW 0.003 13 SSLSpLLLVS E 0.00- 1 MFRVgFLIIS 0.000 SVFUISSSS 0.000 L Table XIII-1 09PI D4v.7 N' terminal-A-10-M rs Each peptide Is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide Is 10 amino acids, and the end position for each peptide Is the start position plus nine.

Pos sorSubseuen j S [1111 FRVGfLIISS l.~Oi 21 VSWrVNTTN 0.000 GLBSSS0. 00-0 Table XIV-109P1 D4v.7 N' terminalAl 101-9ers Each peptide Is a portion of SEQ ID NO, 15; each start posItion Is specified, the length of peptide Is 9 amino acids, and the end position for each peptide Is the start position plus eight VITL PLLLVSWR 0.012 E-1 IYFL _I9 18 LLLVSWRV F666 1 MFRV-10 004 PLLL Sy .003 20 LVSWRVNT[002 5 GFUISJ 0.001 14 SLSPLLLVS 0.001 3 ESLanLL 0x F6 =0.9SSS 01 I_ 11SSS SLP9 10SSSSSLSPLjf =5 LSP ].LVSV =2-L FRVGFLIIS If-jo FI19I LLVSWRVN 00 FT7 ISSSSSLSP iFo0070 .e iIi vFISss_ j1.00J LL YVSRVNTT '0.000] Table XV-109P1 D4v.7 N' terminal-AiiO1-10-mers Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine.

Pos Subsequence j re IT1. jiP Y§ i T.060 iC]LOSSav 1=0 jLyq YR\1N!T][10.002 71RVGF I DoT 6-671i 1=9 10-.LLVV 0071 =YGFLIISSSSS J( O.99~j I !ISSsSSLSPI 0.001 17 LPUL S~RiI~oo Lf IS LS 0.001 Li9 UVY YR!Tf0.0T I= 1 SSSSISPLL ][66 FRVGfLIISS 1.-K E =SS S E6:0:()70, 2=1 11 VSWrVN1TN] U Tb XVIS L109PI D4v.7 L N'termnalA24-9-mers LEach peptide Is a portion of SEQ ID NO, 15; each start position is specified, the length of peptide is 9 amino adids, and the end position Lfor each peptide is the start position plus eight.

Po-sl subsequence, sco-re] I LLVsVRVN F 6.000 Li! MFRVGfLII ]{fi.oo [jl GFUISSSsIL1.00 L VSWRVNT 0.0 f17 1 LSPLLLVSV 10.180 S]LL S 0 1 8 0 00 00 Table XVI-1 09P1 D4v.7 N' terminal-A24-9-mers Each peptide is a portion of SEQ ID NO: 15; each start position Is specified, the length of peptide Is 9 amino adids, and the end position for each peptide Is the start position plus. eight_ 'L~SubequnceT [Score]q- JIRLy" v I I-q.1 -1SSLLVJ 0.140] 1_ ISSSSS11 0.100] =2 Fini-2 0 0-s 1 TaleXIII9PID4v.7 Each peptide Is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide Is amino acids, and the end position for each peptide is the _taoslton plus nine.

j] FLIIsSSSSL 16.000' [I]~ZiSSSLP1 1]48001 1L L PL=L4FOO-01 L]S S LSPL 4.000J [~1Ls~i0.5001 19 LL -SvR=W 0.10 L21 I _Y§W yV=MN 1.210 L 18 J[ U..VsVVRVN Fq100 Fis LPULVw 70 EC3~ -JLcLLJVS 110.1801 [T7.I Fo- TR I5 L.I LPI!IV 1'01441 [-T]VGFUiSSSSlo14o! l[ ili LVVRVi 10120 K2SSSLSPLLLV] 116K LswsY±ll =Table XVIIl-109P1 D4v.7 N' terminal-A24-1 0-mers Each peptide is a portion of SEQ 10 NO: 15; each start position Is specified, the length of peptide Is 10 amino acids, and the end position for each peptide is the .Start position plus nine.

PbsequenceSore 1z1lPLLLvSVVyj 015 Table XVIII-1O9P1D4v.7 N' termnal-B37-9-mers Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino adids, and the end position for each peptide is the start position plus eig;ht.

Ko:] Subsequnc ej ~L!6 SPLIISW 4.000j LgiI:1 sssLsPLLL3(4f00 7= 1 IISSSSS] .00 =io L -SSSNSLSPL JE§K1 D731L ssispwxkv j 3 1 ILkSPLLLVSV :j F iL \C JIV0.2001 f~ i 2 SW1-1 10.100] WLSuiYl[ 00201 WIL I S S S F120.:0:0 DL IsS§9sLP q-0010 GF5 1 FLIISSSS Fl 0.002 F-'1.1i L RVG FLD -6-6 O2j II Table XIX-109P1D4v.7 N' termlnal-B37-10-mers IEach peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start positionplus nine._ -LEijj._. ISSsSLSPL LI4.00o0 Th L SSSSISPLLL 401 JL mVmmn 0.500j F16 1 SPWVSVRIL.200,I V 14 ISLSPILLVSV j 0.2DI L18 L I-VsVViLNoJo2oJ IDT] _S1sp 0.J020J LiiLY~r~il0.020 1 1IISSsSSLSP Dr 001 1= GFLIISSSS 0.0021 Table )C-109P1D4v.7 terminal-833501-9-mers Each peptide is a porton of SEQ ID NO: 15;each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide Is the start position plus eIght J useuec .1 F score] [Th~[SSSSL -FLs-q-o 01 K~~IIJ[y SSSSLSP -I R.9O9 13F-.- 5SLI!V ~1.000 1= VSRV [o ooI 00 00 Table XX-1O9PI D4v.7 N' terminal-B3501-9-mers Each peptide is a portion of SEQ ID NO: 15; each start position Is specified, the length of peptide Is 9 amino adds, and the end position for each peptide Is the start position pluseighLt L?-o Subsequence J[or 167L _vsRrGr 1 0] [i II1LyGFLs t[100 I0 F19 ILLVS 0100 E-i l- LPLPS U7109- D 7 7 L S-LSIFO-100 F '6 I FUSSSSS jF051-0 L EIi a iFsss Io.oo 4 I =0SS 0100 i~L sss-ssLLs:P1 o=65 7 GFLIISSSS 0.010 2 LRVGFu lIoI-Oi1f =7 1j__LULSVR 0v001 Table XXI-109PI D4v.7 N' termlnal-83501-10-mers Each peptide is a portion of SEQ ID NO: 15; each start position Is specified, the length of peptide is amIno acids, and the end position for each peptide Is the I start position plus nine.

[P osjfsuseq 5;cj 9 -I1[ ISSSsSLSPL 5J =6iFJ SSSSsLSPLL FJ 12 1.000 ]I VSVVrVNTTN smLLVSI0.=500 .7:j1 SPLLIVSVVR 10.2001 14 IjSLSPILLVSY 10.00 3 7- RVGFIISSS 0.200i 18 jJLLLsRNJ 00i 19 UjLVSVVRVNT j.iO~ I LVSVVRVNT'3 100 4 1 YGFLIISSSS 10.1001 SLIISsSSSLS Fi i -T MFRVaFLIIS IFOI Table XXI-IOP1D4v.7 N' termlnal-B3501 -lO-mers Each peptide Is a portion of SEQ ID NO: 15: each start position Is specified, the length of peptide Is 10 amIno acids, and the end position for each peptide is the otas position plus nine.

ISubsequence score L D E FRVGfLIISS '!Ej I I jjSsSSSP ~io]0 F lisssss 11001 Table VIII-1 09P1 D4v.8 A1-9-mers Each peptide is a portion of SEQI ID NO: 17; each start position is specified, the length of peptide Is 91 amino acids, and the end position I for each peptide Is the start position plus eight r~JIK us! I1 Score1 II ujLInVQPT ][045j [J F( K=KEI 271 qq F 3 1 IPGLKKEIT 0.003 8 K~rVPT =0-001 WL P TKERV .000 1 GLKKEITVQ 000 Table X-1O9P1D4v.8 Each peptide Is a portion of SEQ ID NO: 17; each start position Is specified, the length of peptide Is 10 amIno acids, and the end position for each peptide Is the start position plus nIne.

Pos I[ Subsequence I[Score I 8Y!] VQP 0.090 L7 II[ PGIkKEIT V 0AP01 7TFIPgU(KEI 1[01 i[STFPGU(K E ][Iij LKaEIVQPT f.000J L5~ PG~kEI1VQ iL0oJ Li GLKeTVQP J0.ooo0 Table X-109P1 D4v.8 A0201-9-mers Each peptide Is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide Is the start position plus eight_ Pos I Subsequence IScorel 2 FIPKEI l"16.599 8 Ql 14.' L -LPG EITV _03? [3 IPGKKEIT ]0.017 7_!1KKETVGIT J0. 0=0 [-57 GU(KEITVQ 0.0001 i TFIPG KEJI 0.000 I 1 LKKEITVQP 0.000 Table XI-1 09P1 4v.8 A0201-10-mers Each peptide Is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide Is 10 amino adds, and the end i position for each peptide Is the startpostion plus nine.

EPos ubsequence Icore I. FIPGIKKEIT~ l .94 L 1[F IPG~tKE[rTVio.o7I iL RKQP 0.022 [21] TFIPgLKKEI EI §fl [2Z]L!ETVQPT IW.p0I [iJ[ STFIpG F0 o.002j KG GLKefVQP 0.j PGU(kEPVQ Y .0j[ Table XII-IO9PID4v.8 A3-9-mers Each peptide Is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino adds, and the end positoni for each peptide Is the start psition plus eight I Subsequence 1[ re [T2[ GKKETrVQ LiI009 8j FIPG KEI 0.045J LT Ti 1 0i 00 00 Table X1I-1 09P1 D4v.8 A3-9-mers Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptidle is the start -osition plus eight ~FTIL rny9T :p-i-.001 1=Y1[ PGKKEIT 00C -Table XI-iO09P1 D4v.8 A3-1 0-mers Each peptide is a portion of SEQ IID NO: 17; each start position is specified, the length of peptide Is position for each peptide is the sta!rtposiion plus nine.

FIPGIKKEIT ILJ i5 FTable XV-109P1 D4v.8 A1l1-O-mers Each peptide Is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide Is 10 amino acids, and the end position for each peptide is the lL F-!ubsequence [re 1ET PG kKE[rViLO-.004 E-T] T FPg 0.001 LIjIII[ KKEltVq TV 10.0 E~IEIT=VQPTVE FO-0000 IA QPT =0-000o S Table XVIA109P1 D4v.8 A24-9-mers Each peptide Is a portion of SEQ' ID NO: 17; each start position is specified, the length of peptideIs 9 amino acids, and the end position for each peptide is *the sta rt positon plus eight.

Pos, -ssln i-F"e [A 1 IPGLKEIT F 0.0] KE8ITV1 P1V 0.0421 I T~J PGLKKEIT 0.151 =5 1~ G LK KE ITV Q I .:1O LTable XVII-109P1 D4v.8] A24-1 0-mers I1 4 L ~k 6F9Yj~L0] If Tble XVII-109PI D4v.8 A24-1 0-rners Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptidleis 10 amino acids, Iand the end position for each 1peptide is the start position plus nine. j [To J[b eue1 core] [iIIIP-GLkKEITV 11jO.1001 WLtV~ffV .0421 LLI1STFILpGJAE10.011 Ei iJ KEITvQPTEJ[O0031 -Table XVIIl-109P1D4v.8 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide Is the start position p us eight.

uencD] Score] L- :UL:fE[ L2. 00 0 F l P- Wi oo: j [iii1_GE I1 jJk)L 0.020 W[ U(TfVQ]IO 1 fTPL KKE j .0 19ID4v.8 Each peptide Is a portion of SEQ ID NO: 17; each start position Is specified, the length of peptide Is 10 amino acids, and the end position for each peptide Is the stat ostio pusnine.

_Subsequence.Iscore] 7F7TPGLKKEIT. 7 ~LKKEr1VQPT 000 F1[1 STFIrtGLKE 1FO50i LLI[STFpGU(E 100041 L ElVqPV p~ Fiii J 9~ LKS i I*] IFi9i11 KEITVQPTVE J0o.000 Table XIV-109P1 D4v.8 1 Al 101 -9-mers Each peptide is a portion of SEQ ID NO: 17; each start position Is specified, the length of peplide Is 9 amino acids, and the end position for each peptide is the start I position plus eight [P os'.1Sbeune~[~ T .IKKEITP IF 00 00 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide Is the L.start position plus eight I Pos L -LL G LKKEI_ 0400 L GtKErrVQil.045 =8 LErnVQPw 0.040 LiLPG!IV_0.020.

51-mers Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide Is the start psion plus nine.

[~[Subsequence JJSccorl 7 1_FIPGIKKEIT 0.100 EKT]L K rV P J.0.~ L7 sZL xErti2I~~ I1 STFpGLKKE g 9 KEITV1P yj[0002 1 PGLKkEITVQ .021 Table XX-109P1 D4v.8 B3501-9-mers 00 Tables XXII XLIX: Table XXII -IO9P1D4v.1 Al-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eighL 91LETGY2 I51 TMQFKLYj 1807) TSDYVKLV i 202 4181 LTAAYLDY 495[ SPNAKNY 1 [594 IVtDPDYGDN 21 985]ISSDPYSVSD11 [3641[VDTLSE 20 F3-511 LE ILTI I IPPSN~SY IJ (789I(SIEAPIPNI F4-116 INGVNY T 351 NPSID!RY 1 [741VDLGLRV I 931 JDSPDLAHY [ml [9811 c~ssssQPY [9 1116 PREIFRLVK JI [21 IADGGLPA 111 345 IVDVP 118 [991 IVDCGYVT 118 IVDGGFQR 11~I [239 IVTDThDINHP [7!1 [251 EIEEVP J1i7 [273 I AIDADIGEN li7l 3jj SIDIRYIVN 17 3851VDD HNW 399 FTDHEIFR 11 528 ILDREKEKY [17j 567 1)SEVT1EY 117 727 DETGNITL [i 929 (KPDSPOAR 17 Table XXII -1O9PID4v.1 A1-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amIno acids, and the end position for each peptide is the start position plus eight [osHIRPVG!QV[] 34 MEENVLIGD 1±i 78 EEDTGEIFT[m [9j R1DREKCA 16 109 EEVAILPD 1 11321 IDNAFP 16 I 3J ADPDVIN W 401 DHEIPFRLR16 531 EKEDKY1LFT 1 631 F E SY 738 KCDVTD GL 802 DVSSPTSDY 1 j897J IDDGNRMTl ft6 11091 IEROEHC.FY F15 LPDEIFLV I 20fl REKT 423 42 )LDYEST~EY 428 SThEY1IK i 591 LrTDDY 16EQESYFY O i645IADGGRXSRI] 6881SNPGTVF15 1705 ITGMNAEYRY 988PYSVSDGY 11j 1 K8IG~DVPIR 14 11481 IENSAINS [I4 211 EKDTY~VMI(\ j 278 IENAIF-F j)4 (3111 IEPLBEE [I4 Tble Xii -109PD4v.I Al-9-ners Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight El I 317 RRETPNL L 319l EIPNHKkLV )O 411 VESNQFE IO 1514 ISDCRTgML j 54 lAKNVPL j (52 (HliENFVP jI~ I612 1Et40FT!DS I~ E 54 1KEDGGVS 1~ I] IIOKPVFVP [71 1681 SELVLEST 17 1720 1(TDLFADQ [71 S758 QIEDSLFW [7 851 NEWATNP [7 TkDLPILE [97PLONTL4AC [71 00 00 Table XXIII 109P1D4v1 A0201-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 9 amIno acids, and the end position for each peptide Is the start position plus eight 18171 AVAGTTVV[4 188o F NLLLNEVTI

I

L6i KLKGDV 3 1231 STALVSV 3071 GUTISEPL 11LAKDNGV23 [745] GLHRVVKA J 810 YVKILAAV 2 13~ ILVAAYAGTfI L3Ii VUGMIGtLKD J 741 VTD3LGHRV J 816 MAVAGIITV W IFAVI4ACV 2 76 (1RIEEDGEI 2 124 KIRFLEDI 2 152 SAINSKTL 2 301 HLNATIGLI 2 a1 DIRYIPV[2 360 IVNP~fDTV [1 $536 YLFTJ[21 743 DLGLHVLV2 820 GTITWVVI j* WVIFITAV 21 jso II SLIPNSLT Lo 127 FUEDNDN 23 ILQVSTDT I 270 QLHATDADI 2 Ij9I RLFHLAT I [33] LMPARMV 20 337 1ARAMVLVNV 2 34] MVLVNVTDV 2 347 DVNDNMPSI 2 F359YIVNP~NDT 1421 STKEYKL 2 Table XXIII 1O9P1D4v.1 A0201-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position pluseght.

54 GVPPLSNV 20 550 LTSNVTVFV 20 656 SAKVTIN 20 658 KvTINMDv [I] 715 IVGGNIRDL j 725 AIDQEGNI t~jTNATLINEL 78111 LINELRKS

$IP

826) WIFITAW 20 Lii GIYIFL 19 LACWFH 19 L2 iI GAQEKWITI W91 135 NAPLFPATV 1 162 MAVDPDVGI 1 303] NATTGLITI 1 326 LLASDGGL 19 1377) NTK(IAJLITV 19~ 1438 AADAG!PPL L9 (50-31IGPAPP 1 5421 AKDNGVPPL 19 5831 LPRHGTVGL 616 818al VAGTITVV 19 8811 LLLINTIE 19 903] VTlEiDL 19 914 QTMGKN 1 W LSGTIFA 1 IILSGYFAV 18 S13 )ILLACVFI-S JI1 51 )1LIPNKSLTT 181 Ijr KLCAGPRD 18 11201 FRLVKRFL 18 1211 RLVKIFLI 18 213 1j 18 276 ADIGENAKI [263J KIHFSfSNL 1!§J /Table )(III 109P1D4v.1 A0201-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eIght.

F I 3691 VLSENIPLN jj 381 ((ALTVDKD [I1 4031 EIPFRRPV 78011 SPGIQIV $ij 1496 11GPNAKNL Ij~ [Ijo ll DENDFT $jI1 617( TIDSQIIGVI Ij 1693 11TVFQ~AV $jHJ S7331 ITLME~CDV [I] 1734 ITLMEDVT I] 57481RVVKNDLI] 757 lGQPDSFSV 17621 IFS VIVNL JEH [7801 TLINEVRK 18141 LVAAVGTI (I~ 18221 ITVWIFI I~ 19 I1 PNSKHII [95jJ SKHHI!QEL j)J 990 1svsDc~Pv i I8E YIFAVLAC J [31 LAMFKL W71 88 (si GARIDEEKL j(J [i*3)1 VINISPEN jjO 1561 SKYTLeAAV ~I] 165 JDPDVGNGV 1] 1791 IKSQNIFGL [E] 2561 VSIPENAPV 1320 I TPNHIL (Iil S327 VLMASDGLM 1] 368 WLSE!IPL 13791 KIAlIJTDr II i482fl GQLTVSA I 493 1ADSGPMKI 61 HGTVGU1V jO I6 VLIPSTPGT 7611 SLFSVVNI11 00 00 Table XXIII 19P1D4v.l A0201-9-rers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptde Is 9 amino acids, and the end position for each peptide is the start sition lus eight 882 LLNFVIIEE 116 19341 DLARHYKSA 081 HTRPVGIQV 16 41 GDLLKDLNL 15 i1 UAMQKLV Vj [ma Ij LPMAV~PDV [Ps F61 INGVQ 181 SQNIFG EV 5 182F 3NIFGDVI 15 229 RSSTA!LQV Ls [F3 PVGTS96QL] II1FSFNLV [s I2I[ SFSNLSNI [s 338RAMVNVT [3Z4 IPL~TKIAL 1 396 VTCFDHEI15 F6 QSAMFIKV 15 8 AMLFIKVKD 15 [451 MLFIKKDE i! Ff 15GPDAPPE 517 CRTGMTVV1 590 GLIVDPD 1 624 V1RPNSFD 643 1VKAEDGGRV 1 651 VSRSSSAKV 68 STNPGTF 1 707 MNAEVRYSI15 742 TDLGFRVL g1 767 IVNLFVNES 1 769 NLFVNESVT 1 875 KHSPNLLL 1 897 D1 SDGNRvT I15 TLDLPDLE 15 [0J DLPIDjEEQ 1 Table XXIII 1O9P1D4v.1 A0201-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 9 amino adds, and the end position for each peptide is the start position plus eight 1961 HIIQELPLD ifsH 970 NTFVACDSI [9]l L4IJ LKDLNLSLI [4 [ij6 DLNLSLIPN 66 IVYKGDVPL [4 [111 EVAILPDEI 14 113 AILPDEIFR 14 115 LPDEIfRLV 1 128 LIEDINNA 1 137 PLFPAIVN 1 138 LFPATVINI 1 [1zI SIPENSAIN 1 [59ITnPAMPD [4 183 NIFGLDVIE jj 211 EKDTYMKV J~ 232 TAILQSVT 248 VFKETIEV 250 KETEIEVSI 31 TIKEPLORE 132IIKLV9SDG 1329! IASDGGMPA 339 AMLVVTD 34 J NVDVDNV 1 362 NPVNDIVL L3AJ KDADHNGRV [412 FSNQFLET 4 VFTQSfVT jN SOO] KINYLGPD j4 5071( PDAPP~FSL 14 516! DCRTGLTV 1 5401 ILAKDiNGVP [4 00 Table XXIII 1O9P1D4v.1 Table XXV- Table XXV- A0201-9-mers [109P1O4v.1-A3-9-nersj 109P1 D4v.-A-9-mers Each peptide is a portion Each peptide is a Each peptide Is a of SEQ ID NO: 3; each portion of SEQ ID NO: portion of SEQ ID NO: start position Is 3; each start position is 3; each start position Is specified, the length of specified, the length of specified, the length of peptide is 9 amino acids, peptide is 9 amino peptide is 9 amino and the end position for acids, and the end adds, and the end each peptide is the start position for each position for each position plus eight peptide Is the start peptide is the start Sposition plus eight. position plus eight.

552 SNVTVEVSI 14 rI 571 THNEYNFYV 14 780 TLNELVRK 28 [1 IVF3P6t1TV 6781527 KLDREKEDK 26[748] RVLVKANDL 00 [686LPSTNPGTV 14 172 GVNYELK 24 18261[ W!FIIAW 407 IRLPVSNQ 124 J [iii WEHSQE jII 0 14 F4-0-71 5901 [NPG-AwFoV-0 F8-1JvErv j4 [nf PDEIF&LV _HSG 9] C-I 706 14GMNAERYS 1 714 SIVGGNTRD8149 1 APHLKMI(24 [1891 VIETPEGDK 768 VNLFVIAESV 14 f AYkDYESTK 23 218 77] NESTATh jfJJ !674 1)IVEPSNSY 12311( 220KgD EQ [81 ELVRKSTEA 41 )(841 HLAA~NK 234 ITVDKADH 1 812 KILV4VAG 14 972 38CDSISK 23 14161 FLLETYL Ii11 878J PKNLLNFV 912 VLg2ACWFH 433 AI 9DA 11 DVDSDGNRV( 233 1 AILQV:79TD ]2 [141 NSEGITK Ii 941141 518 RT2ML2VK 2[j KYFTLLAK 19 948] FIQETPL [1 11623 lrGVIRPISF 22j 1549 ((PLSN~{VF[M r662 NVDVNDNK 22 1 E9GITVTD1 TableXXIV 814 gLVAVT I [2J 665 DNDNV F 109P1D4v.1 F8 -33 CR] APH 22 802 DVSSPTSDY A0203-9mers r56 SLFK [1 2 D1SG1YIF (81 [65 jLKrGQVP [21 [j M GDL!KD 1 No Results [6 DIV 28 Found. Fl-6-71 219 298 RLH WlT 21 90 RIHRECA Table XXV- 324 EfLLAD 21 21218 109P1D4v.1-A3-9-mers F379 18A~ffD 211 SVIQCjT D Each peplide is a 1524 V1lKLEK 211 F3 331 portion of SEQ ID NO: 3; each start position Is 1582 1NLRHYG 11 445 1 QS LF specified, the length of 740 DV8LGHR 21 407 peptide is 9 amIno __FIE__ 74-41 R~K 21 1814 IV acids, and the end 2a] position for each 812 KIVAAAG 21 642 1YVAEDGR 1 peptide is the start 1817 F6AVAGTL i I 645A position plus eight WVTTPFK 21 688 STPGYF RV01 RSSSAK 31 .j 18 2 435 1Nfl 18G 113 AILPD1FR (20 [697 fQVflV4LDT l 11 IAV1JAVVF 2 37 NLIGII llRl197 KMPQWLQK 119f745 GLRWYKA f 00 00 Table XXV- 1109P1D4v.1-A3-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amIno adds, and the end position for each peptide is the start 1position plus eigh 1832AVIRC 1AP L8 F83 RC QA [18 871 RK19(K8SPK L~ I1002 EvEvsLUR 8 1006 SVjTRGI 3 I KDL~SL [7I 951 KL1AGIPRD17 1221 LVIRFLIE 17 137 PLE7PAW1N 17 1631 AE7PDVIN 17 177 ELKS9F 17 2101 EED K17 2571 SeENVG 17 270] QLEAT7DI f1i 290] NLSNIARR 17 3811 AL7EKD 17 W6] E7 436 17 48] QLV~qMD 17i 503 YLGPDAPP 17 604] 'AVILSlE17 624) V1PNFD [7E 710 1 E7VG 7551 DL EPL 7 [L 76W1(VN!~N[7 7691 NLFVNVT TJ ATN~VR E7 8131 IL17EGT l 8211 TIIWIF [7 1013] G1qVS! TF 17 [6 73i[ PL!RIEDT [61 741 LIIE TG l 1311J DIDNALF Le 20111 MQKDR 16] Table 1V- 1 09P1D4v.1-A3-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amIno adds, and the end position for each peptide is the start position plus eight IE 6 2I :SVIDNH16 i 242J 1TNDNHEFK [6 a77II DIENAKIH [161 :93:S4A E 30 fl AU~GLIIK [e~ 3411 E~~V 16 1 351] E6IIR I 371 1SE6IPTK )e 38D IAIIDK [16 4491 SAMLFIVK 16 5041 E1PD6EPE [6 54:GVEPLV E so fl SIjDEfDDF [6~ 63II E6I 700VNDLMN 1 713 YSIVGTR 16 734 TLMEKVT 16 743 1(DLGLH~LV 161 750 ILVIANGQ 16 761 SLSVVN 16 764( SWIV ~FV 16 810 YVIjAV 1 934 DLRHYISSA 16 96zJ PLDNAC Table XXVI- 1O9PD4v. 1 A26-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide 1s 9 amino adds, and the end position for each peptide Is the start siinplus ei h

I

[802 11DVSSPTSDY [65 IDVNDNKPVF I8 2411 DTNDNPVF I 61 ENVUGDLL [I [IEVEVAPD 3471 DVNNVPSI [1J 10021EPVSVHTR 11501 ENSAINSKY j(J 188 )1DVETPEGD j1 351 NVPSIDIRY [422I PVSNQFLL jIJ [2jJ[ GV1RPNISF 11j4 [F4h[EVRYSIVGG [ilh EIFRLVKIR I 21 ETEIEVSIP (3 2631PVGTSVTQL [741 DVTDGLIR ~Jj 131 i DINDNAPLF [I] 177( ELIKSQNIF (22] 419 11ETMYLYE [O 477 ENNSPGIQL j(j f3 EKQESYTY 674 PPSNCSY jJ 1729 ))ETGNITLME 1J S71 11DVPIRIEE j~J ~I DTGEIFTTG ft0 111 E AILPOEI jIj I167 (DVGINGVN (O 9191 JEPEGDKP 1 255 EV RSIPENAP 2801 ENAKIHFSF I [18 J EETPNHKLL 0 [42][1 STKEYAIKL 1 693 )1VWFVIAV [I] 8061 PTSDYVKIL 1993 1)DCGYPV~TFJ LVSNIARRL WLSENIPL I [9i ][DHNGRVTCF o] a][w VVKoURE F2551TwVSllDo f aJ IDVDSDGNRV J 00 00 Table XXVI- 109P1 04v.1 A26-9-mers Each peptide Is a portion of SEQ ID NO, 3; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eiht [931 OSPDLARHY j0: j3 1 EIFTTGARI 19 [218 KVVDG i 319 ETPNHKLLV9 F53 LV1ASDGGL9h F751-51 E19I i 748 RE9VKADL 765 WIVNLN 19 809 DKLVAA 19 823 TWVIFT 8251 VFITAV 1 903 VTLDLPIDL L 953 ETPLNSH 9 11AVLACWF 18 33 EMPENG 18 [39 LIGDLLDL 8 FlLT8AMQFKL 141 1 E18IP 142 N R8IP F253 EIEVSIPEN 3 6 DIRYIVNPV [I 403 EIPFRLRPV 458 E8DENDNAPV 56fl

F

570 E18NP F6881HSTNPGTWF 8 F694I E18vvDI lZiIlDEGNITh [8~ 7631 E8INL IZ6I FSWIVNLF [82ll TIIWVIF j \N WIFA E8~ F8I[ 1 8 F8-2-41 Rvosl Table XXVI- 109P1 D4v.1 A26-9-mers Each peptide Is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 9 amino adds, and the end position for each peptide Is the start position plus e F89 7DS1GNRVTL DLLSGYIF117 E DEIFRLVKI 117 [13 [DTYVKVKV I117 3 DNVPSIDIR 17 372 ENIPLNTKI 17 413 EYAIKLLAA 17 [7J IYVPENLPRH 1 [81 GVGLIT 17 704 IDTGMNAEVR 117 17 [LGQPDSLF 7 1899] DGNRVTLDL WI LII 1 GTYIFAVL 16 16 lCWFHSGAQ 16s 17 1WFHSGAQE 16 79 EDTGEIFT 163 AVDPDVGIN g 29 NIARRLFHL 6 (529 REKEKYL1 S53 NVTVFVSII 116j ~IATLSILDE 16 614 1 DDFTIDSQT 16is F658 KVTNVVDV 16 659 VTINDVN 16 764 SWIVNLV 16 771 I FVNESVTNA 16 791 EIADVSSPT 16 810 VILVV 16 I DSISKCSSS I~h MJ TTFEVPVSV 211 EK1 Table XXVI- 109P1D4v.1 A26-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus eight 277 DIGENAKIH 1 3201 TPNHKLVL 1(151 j~jM VLVNVDV 11 363 PVNDTWLS I 367 TWLSENIP 470i~ FVTSIPEN )O 47 V1 SIPENN 549l PLTSNVVF 1 567 SPvFINEY 591 LI1T9DPDY 605 vTLEN E1 [9 662 INWDVNDNK ji Kil EPRIPS 15 LfIPVIVPPSN i F-n1i l 1 71784 [V ELVRKSTE71 832AWRCRAP E1 1 [E IRQMIMMK [5nJ SPKNLLNF E0 [ss][VIEETKAD Ifs r902 1 RVTLDPID I~h [958I SKHHIIEL 110111 PGI~VSNT I STable XXV1I-10P1D4 Sv.1 -B0702-9-mers Each peptide Is a porion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start poiinplus eight 1 583 ILPRHGTVGLE 583111RHGE15 00 00 Table XXVII-109P1D4 v.1-B0702-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus 2!ght 1362 FlNPVNDTVVL IE2 F36HAPFPAT 23 F374( IPNTKIA 22E [09 J RPVFSNQFL f~ [676 PPSNCSYEL 2 [792 1 APVTPNTEI j~i 441 PPLNQSAML21 496 2GPNAKINYL 2 4041 IPFRLRPVF 20 52 IPNKSLTTA 19 160 LPAAVDPDV 19 258 IPENAPVGT 19 3351 M PARAMV 19 5463 EAPVTQSF 19 S758 IQPDSLFSW 19 115 LPDEIFRLV 18 226 1 FPQRSSTAIg 13521 VPSIDIRYI 18 443 RKPPLNQSAM18 4751 IPENNSPGI8 F 40]1 SPGIQLTIQI S548 )1PPLTSNVV 1[8 686 I LPSTNPG1V [a 690j NPGTFQV 18 541 SPTSDYVKI18 877][ SPKNUNF 18 6901 RKPDSPDA1 9LPLDNTFVA 18 ]fDPDVGINGV 17 8771 HPVKETEI [f7 547 VPPLTSNVT 1 596E DPDYGDNSA7 7951 TTEIA7V 117 NPENRQM11 Table XXVII-109P1D4 v.1-80702-9-mers Each peptide is a portion of SEQ ID NO: 3; each start posItion Is specified, the length of peptide is 9 amIno acids, and the end position for each peptide is the start position plus elh 262 APVGTSVTQ [6 4jj JAADAGKPPL [6 493 HADSGPNAKI [6 506 HGPDAPPEFS1 542 1AKDNGVPPL [1 82] 9NPENRQMIM[6 875 K SPKNLL [61 897 IDSDGNRV [6 9071 LPIDLEEQT [6 95ILTPNSKHHI_[61 I ]fEMPENVLI[I 4ZII) iNNSPGlQL I~ 5i0 1PAPPEFSL jIO 71511INR L( 10,01 FQIQPETPLI5 101011RPVGIQVSN Ii l I~IIPRDEHOFY L41 15411 NSILPA E41 227 1 PRSSTAIL [4 317 E1REETPN4KL[4 5jj (APPEFSLC 14! 670] KPVFIVPPS (4 7381 KCDVTDLGL 14 76:WIVNL F4 S Jf SGTYIFAVL1 i4i If LSLIPNKSL [6] js I[WKTGDVPL I 68 1f GARIDREKL Ift~ 13011 EDINDNAPL 162 E3VDPDVGI 1 1791KSQNIFGL 13 192 ITPEGDKMPQ [6 263 IPVGTSVTQL [3 :1EDKYLFL [3 [Table XXVII-109PI04 1 v.1-B0702-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 9 amino adds, and the end position for each peptide Is the start osition lus eight I I YGDNSAVfL [6] [ZISNCSYELV 1 [74I TDLGLHRVL 17 NESVTNATh [s11 JPTSDYVKJL I 181H IIAVAGThW Iii 1839 APHLKMAQK [6 I9IIDGNRVThDL I11 fl 9KSASPQPAF 3 8951 IIE3PETPNSK 196011 HHIIQELPL 13 ITable XXVIII-109P1D4 1 v.1-B08-9-mers Each peptide Is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus eight.

496 1GPNAKINYL l~ F43 LELNLSL 320 1)TPNHKLLVL I~ 453 FIKVKDEND 514 2 2j IGAQEKNYI 246 HPVFKETEI 428 STKEYAIKL 877 SPKNLNF 1201 FRLVKIRFL I 2123 [3sJ PLNTKAU [19 EDKYLFTIL LPRHGTVGL 3 00 Table XXVIII-109PI D4 v.1-808-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eih 41 GDLLKDLNL 66 YKTGDVPLlg 294 NIARRLFHL g 955 USHI 88 I(GARIDREKL 736] MEKCDVDL 748 RlVKANDL 8662 8672 868 KKKKKKKKH 869 EMQOIJ21S 873 KKKSPKNL 875 KSPKNf L I l IDREKLCAG 1193P 20 F8- 5KKQS 20 870 11M J20J 871 KKKK 20P 927 TFKPSPDL0 416 FLTMYL1 631 FDREKQESY 784 ELVRKSTEA 1 114 ILPDEIFRL18 122 LVKIRFE 18 334 LMPARAMVLI18 374 IPLNTKALE 451 MLFIKVKDE 18 5281 LDREKEDKY 1 530 REKEDKYLF 1 656 SAKVTINW 666 VNDNKPVFI18 734 ITTLMEKCDVr 1 F741 64KLVYKGDV [72 VPURIEEDF7 Table XXVIII-109P1D4 v.1-B08-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amIno ads, and the end position for each peptide is the start position plus eigt [124 KIRFLIEDI ff17 218 KVKVEDGGF 17 307 GLITIKEPL 1 7 362 NPVND1VL 17 RPVFSNQFL 17 I426l YESTKEYAJ 676 PPSNCSYEL 8391 APHLKMKf7 jIjo~s SVHTRPVGI ff7 1152)1 SAINSKYL J[i6 176 YELIKSNI 16 )227] PQRSSTAIL 16 310 I TIKEPLDRE 16i 313 EPLREETP 16 405 PFRLRPFS 16 31 E6PPLNQSAML1 I 633 1REKQSYTF ff6 13KMQKNKQNft~ 3] LIGDLLKDL fi0 [1[1 DEIFRLVKI fiO i 391 IDHNGRVTCF I~j 433 1AKLLAADA 15 541 LDNGVPP 15 E3rsfl 5 I e3] IWROROIAPH [O 864f IMIMMKKWKK1 Sill LIPNKSLT ff4 !!rI IFRLVKIRF ff4 131 AINSKLP 4 ~Il INGVQNYEL ff4 177 ELIKSQNIF 14 201 LIVQKELR 1 203 IVQKELDREE 226 FPQRSSTAI Table XXVIII-109P1D4 v.I-808-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position plus eight 2481 VKETEIEV f4 281 NAKIHFSFS F283 KIHFSFSNL 14 13081 UTIKEPL I~ 352 VPSIDIRYI ffj SIDIRYIVN ff EIPFRLRPV f4 [3](AADAGKPPL fj4 4 NAKINYLLG pep[TILAKDNGV [0J SDYVKILVA 4 [8581NE QMM 4 880 NLLLNVI1 f4 9581 SKHHIIQEL f4 Table XXIX-0 P1D4 v.1i-B1 51 0-9-mersJ Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino adids, and the end position for each peptide is the start position plus eight 875 23 3Th0I FHLNAGL f0 960 HHIIQELPL 391 NR C 1 114 ILPDEIFRL g 179 IKSQNIFGL 715 IVGGNTRDL 742 TDLGLHR 1 897] 291 LVSNIARRLj 00 00 Table XXIX-109P1D4 v.1-B1510-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide Is the start position plus eight 4 TDHEIPFRL [s 7621 LFSWV1VNL 5 31 REEMPENVL [14 104 EHC HVEV 120 FRLVKIRFL 170 INGVQNYEL 4] 1318 IEETPNHKLL i4 3621 NPVNDTVLI 374 IPL KIA 1 401 DHEJPFRLR 1 507 POAPPEFSL 1 599 YGONSAT 14 777 TNATLINEL 4 927 TFKPDSP[ SGTYIFAVLL J1 E6I VYKTGDVPL 13 107 FYVEVAIL 1 (193 PE KM L 3 (4j NHPVFKEE [3 (2(JTPNH(LW[3 42 TKEYAIKLL 4 IJAADAGKPPL 1 542 IAKDNGVPPL 113 ILPRHG1GL 1 688 STNPGTWF 113 7 27 DQ ETGNT 13 [740] LHRVVKAN 13 [n7][NESVTNAmIi PTSDVKIL J3 [li SGTYIFAVL(1 F-6- 35PENVUGOL[2 [2][GARIDREKL_2 F7 SASKYTLE 284 IHFSFSNLV 12 Table XXIX-109P1D4 v.1-Bi510-9mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide Is the start position plus eight 307 GLITIKEPL 112 317 REETPHKL 322 NHKLLVASI12 334 12 404 IPFRIRP 1 477 ENNSPGIQL 1 496 GPNAKINYL 1 497 PNAKINYLL 1 520 [NIL] 12K 529 DREKEOKYL 1 [571 I TNEYNY [2 575 IYNFYVPENL 2 [602 INSAVTLSIL g r665 I DVNDNKPVF 1 676 PPSNCSYEL 12 678 SNCSYELL[I1~ 754 NLQ SL12 (874 KI<HSPKNL jI2 19031 VTLLPIDL1 94] FQIQPETPL 1 19g51 SKHHIIQEL 12 110071 VHTPVGQ 12 Table XXX- 109P1 D4v.- B2705-9-ers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptlde Is 9 amino adds, and the end position for eachpetd is the start position ptus 1201 FRL IRFL Table XXX- 109P1D4v.1- B2705-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight F94 GRVTCFTDH 5291 DREKEKYL (861 NRQMIMMKK k0IILRPVFSNQF 625( IRPNISFDR 1316 DREETPNHK 83VRCRQAPHL J I41 1 GDLUCDLNL E 92a I DREKLCAGI IRKCc I (197iKMPQUVQK

IJ

6 REKQESYTF 1901 I NRVT LDLPI j~ (47 I1 LNLSLIPNK I 3041 ATTGLITIK 520 GMLTWKKL 58j4 PRHGTVGLI j 1623( GV1RPNISF (~J 7481(RAM 748 RVLVKANDL 175( IRIEEDTGE Ii ]177I EUKSNIF Ij~ 297l RRLFHLNAT [IJ 317 IREETPNHKL jI] 496

GPNAKINYL

535KLFfLAK LI G1YIFAVL S31 REFEMPENVI F1 9 55 KSLT4 AM1FO 11141 ILPDEIPRL Ii0 119 IFRLV1(IRF [17 290 NLVSNIARR (7~ 307 GLITKEPL 7 I1 KEPLR f7( [35J IRYIVNPVN 11 00 00 Table XXX- 1O9P1D4v.1- B2705-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus eight.

404 IPFRLRPV 409RPSNQFL1 479 NSPGIQLTK F7 [518! RTGMLTVVK [17 REKED C'F J1 649 GVRSA1 6501 RVSRSSSAK L 1747 HRVLVKAND 17 762 LFSWIVNL 1 780 TLINELVRK 1 885 IMMKKKKKK 1 94J FQQPTPL 1 f61 QELPLDNTF 1 11 IAVLACWF 1 [i3i] NVLIGDLLK 16 125 IRFLIEDIN 16 152 SAJNSKYTL 16 179 IKSQNIFGL 1 199 PQLIVQKEL 1 IREEKDTYVM 16 221 VEGFQ 6 1 2711 ADIGENAKI [j6 W8 KIHFSFSNL 1 337 IARAMVLNV1 DNVPSIDIR 1 380 IALITVK 1 435 KLLAADAGK 1 [5i7]CRTGMLTW 16 [71 YSIVGGNTR 1 742 TDLGLHRVL 16~ 777 TNATUNEL 1 827! VIFITAVVR 6 Table XXX- 109PID4v.1- 82705-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus eight 4161 FLLETAAYL E 422 A DYESTK E j2I STKEYAIKL 438 AADAGKPPL E 445 PLNQSAMLF 51 449 SAMFIKVK 4971 PNAKINYLL [5Th]TGMLTWKK 1 524 WKKLDREK SAKDNGVPPL E1 57][FPENLPR 66(2NWDVNDNK 1 F688STNPGTWFIlI 728 DQETGNITL SQETGNITLM r744 LG-IRVVK 754 NOLGQPDSL I] 755 DLGQPDSLFfj 7 ATUNELVR1 82 GTIWI 863 1QMIMMKKKK[O 873 KKKHSPKNL )O 8941 KKHSPKN 877I I SPKNLLLNF [1] (894 DDVDSDGNR1IJ F9-291PDA( KPDSPDLAR jJIAYSASP 958! SKHHIIQEL 1DlCG n VFSGAQEK j~4 LI GAE(NYI j1 1I~i KNYTIREEM I4] 30jIREEMPENV [O [11 PENVUGDL 00 00 Table XXXi 09P1 D4v.1- B2705-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 9 amino acids, and the end posiion for each peptide is the start position plus eight 43 L s-j4O-NL 57 LTAMQFKL AMQFKLVYK14 66 WKTGDVPL1 68 KTGDVPLIR 121 RLVKIRFLI 130 EDINDNAPL F36APLFPA 1701 INGVQNYEL 11721 GVQNYELIK 14 212 KD1YVMKVK 218 KVKVEDGGF 280 ENAIHFSF 14 291 LVSNIARRL 14 E0 FHA0TGL 4 320] TPNHKLLL4 326 LVLASDGGL14 371 SENIPLNTK KI TDHEIPFRL14 14271 ESTKEYAIK 1 I54J KPPLNQSAMI14 44 1 PPLNQSAML f 48 IQLmVSA 1 fl ADSGPNAKIH [27 KLDREKEDK 1 [4J PLTSNVF 1 5991 YGDNSAV 14 6081 SILDENDDF14 618 IDSQTGVIR 627 PNISFDREK14 711 VRYSIVGGN14 KCDVTDLGL1 Table XXX- 109P1D4v.1- B2705-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amIno acids, and the end position for each peptide is the start position plus eight [7631 FSVVIVNLF 11z SSSPTSDYVK 14 83611 CRQFPHLKA 14 8451 HLKAAQKK1 86 ][MIMMKKKKK I4 [~]DSDGNRVL ~J IvTLDLPL I4] F51I[QPETPLNSK ff4] ~][PETPLNSKH I4] SGTYIFAVLIi 36] ENVIGDLL 13 59 ITAMQFLW 13 8IFTTGARIDR 13 87 (TGARIOREK 13 89 IARIDREKLC 13 ~I EKLCAGIPR 13 j~j GIPROEHF 13j 14JJ1 ISIPENSAI 13 ENSAINSKY 1 ~iI IETPEGDKMn 13 j93j(PEGDKMPQ. IO ~ZI DADIGENAK [i0 2~ IGENAKIHF I131 315 11 LDREETPNH 11 33j LMPARAMVL F3 351 INVPSIDIRY 13 36J NPVNDTL L3 4151 QFLLETAAY 13 424 LDYESTKEY 13 429 TKEYAIKLL 458J DENDNAPVF ~4ENNSPGIQL Table XXX- 109PID4v.l- 82705-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 492 DADSGPNAK [3 507 PDAPPEFSL1[ 533 EDKYLFTILH 562 (DQNDNSPVF 13 15691 VFNEYNF 578 1 PENLPR 1583 LPRHGTVGL13 63 FDREKQESY 632DREKQES 652SRSSSAK/T 653RSSSAKVI 1 665 DVNDNKPV 16741 IVPPSNCSY 1 Sg1PPSNCSYEL 3 699 HAVDNDTGM (i I IVGGNTRDL 720 TRDLFAIDQ 730 TGNITLMEK13 736 HEKCDVTDL 773 NESVTNATL 1 792 APVTPNTEI 821 TTWVIF WA4TPNPENR 884 NFVTIEETK 921 WV1TPTFK 927 TFKPDSPDL 930 PDSPDLARH 960 HHHQELPL 972 FVACDSISK 102EVPVSV-TR fl 13 8209-ersrs 182 00 Each peptide is a able XXXI-IC portion of SEQ ID NO: B2709-9- 3; each start position is apetI specified, the length of Eacf pepU peptide is 9 amino portion of SE, adds, and the end 3; each start I position for each peptide specified, the V')Is the start position plus peptide s 9 acids, and t! position for ea( is the start po 120 FRLVKIRFLJ J eight, C1 834 VRCRQAPHL 2 337 JARAMVLVNV 21 76 RJEED1 I IREEMP ENV j1] 102 RDEHCI 250 1 00 529 DREKEDKYLI20250 901 NRVTLDLPI[J KI2-IFSF S408 LRPVSNQF 19 291 LvsNIA 517 19CRTGML19 ARRLF 584 PRHGTVGU 19 362 NPVND1 786 VRKSTEAPV 1 68 19v 36 92 7DREKLCAG I 8 4 208 DREEKDTYV 18J 406 FRLRPV 6] G1YIFAVLL 17 410 PVFSNQ 41 NGDJDLNL 7416 FLLETA 748 RVLVKANDL 49 GPNAKI RRLFHLNAT 16 542 ADNGV 520 GMLTVVKKL 16 546 GVPPLT 307 GLTIKEP1 5 575 FYN11Pi 409 RPVFSNQFL 1 [IIi REKQES' 649 9GRVSRSSSA IS 658 KVTINV 711 VRYSiVGG 5iS 718 GNTRD 31 I REEMPENVL 4 738 KCDVFD[ 1 KSL1TAMQ1I41 R873 88 GARIDREKL 4 874 KKHSPK 121 RLVKIRF 14 9 TFKPDSF 125 IRFLIEDIN 14 2 DLLSGf 209 REEKDTYM 14 5 SGYIFA 229 RSSTAILQV 14 11 AVLACV 317 REETPNHK 1 22 gfl GGLMPARAM I.7 J 136 ENVLIGD 357 IRYIVNPVN 4 49 LSLIPNKI 394 GRVTCFTD)H 1r67 IYKTGDVF 530 REKEDKYLF 4 75 IRIEEDTG 653 RSSSAKVI 14 F89 ARIDREKI 820 GTITVVI 14 [99 GIPRDEH 875 KHSPKNLLL 14 114 ILPDEIFR 26 L0KNYTIREEE 13 EDINDNAF 00 00 Table XXXI-1O9PID4v.1 B2709-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide Is the start position plus eight 960 HHIIQELPL D 2 43 L DLNLSL 1 57 1TTAMQFK-L ii 64K[WKGDV J F66 IWKTGDvPL IEl F8 EITGARI 1 106 CYEVEVAIl 107 FYVEVAIL 11 j140 ISIPENSAI i J162j MVDPDVGI li 176 YELIKSQNI 11 179 IKSQNIFGL 1 190 IEPEGDKM iI [213 DTVKK 22 1 PQRSSTAIL ~IJ 320 TPNHKUNVL 1 334 II.1I ii 34 NMVNVTDV 11 353 PSIDIRYIV 11 42 STKEYAIKL 1 457 KDENDNAPV 11 507 PDAPPEFSL ~1 540 PPLTSNVTV 1 549 PLTSNVTF 1 569 VFTHNEYNF 1 F1 ENLPR62 G PDYGDNSAV iii [62111 QTGVIRPNI Iii [3J KQESYTFYV Jul 676 PPSNCSYEL 1 715 I/GGNTRDL 1 [no] TRDLFAIQ 1 7331 EKCDV ll Table XXXI-19P1D4v.1 82709-9-mer Each peptide Is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position plus eight.

757 GQPDSLFSV 11 763 FSWIVNLF 11 806 PTSD EKIL 1 821 TIWIF 822 ITWIFI li F836 CRQAPLKA 1 86 1NRQMI MKI F8801 NLLFVI 11 D 9 IVDSOGNRV [IJ 187 IDSDGNRVL L 971 899 HGNRVTLD1 J ARHYKSASP 11 L~z ASPPAFI 11 FQIQCPETPL 11 958l sKHHQL 11 964 Q1 ELPLDNTF 11T 983 SSSSDPYSV 11 995 11 9 TTFEVPVSV i 1013 GQVSNTF Table XXXII-109P1D4 v.1-B4402-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position Plus eight.

EETPNHKLL

32 EEMPENVI 964QELPLDNTF 117 DEIFRLVKI 2 58DENDNAP Table XXXII-109P104 v.1-B4402-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino ads, and the end position for each peptide is the start position plus eight 35 PENVLIGDL 23 37REETPNHKL 23 73NESVTNATLn 1 REEMPENL 26YESTKEYA j KEDKYLFTI2 SIEEDTGEIF 21 250KEEIEVSI 418 LETYLDY[]

I~JREKEDKYLF

I]REKQESYTF J

MEK(CDVDL

1176 YELIKSNI [I HEIPFRLRP E81 [ii AVLLACWF 1! 372 ENIPLNKI 1 ~JSTNPGTWF 7j~ 75KHSPKNL 7j~ 82GIFTA [JEDINDNAPL 152l SAINSKYTL I'i BIJEIKSQNIF J~ 26ADIGENAKI iI 49TKEYAIKL Ui~ 52!GMLTKKL 1 54JAIDNGVPPL j 709 AVRYSIVG 72!QETGNITLM i1 SDSDGNRVTL EGI 36 ENVIGDL 5:20KSLn4A] 00 00 Table XXXII-109P1D4 v.1-B4402-9-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 78 EEDTGEIFT 1 1141F ILPDEIFRL 120 FRLVKIRFL 9 [i2 IEDINDNAP 5 [i510ENSAlNSK' F 18VGINGVQNY 15 17 IKSQNIFGL L1 125KELDREEKD [s1 21LVSNIARRL Ws0 GUTIKEPL 15 32NPVND1VL 1 34 IPLNTKIAL 15 SIPFRLRPF 15 415 QELLETY [51 ~59YGDNSAVTL[s 63GVIRPNISF 1

LFSWVNL

TNATUNEL 15 861PTSOYVKIL i1 (8GTI1VVVVI (8JNLLLNFVTI F3912 EQMKNjs Table XXXIII-1 0P 4 1 Sv.1-85101-9mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start pitoplseight 136 APLFPA1VI I)I7

GAQEKNYI

M599 EKMIIIJ Table XXXIIII-109P1D4 v.1-B5101-9-mets Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start psition plus eight [Th-][CFYEVEVAI 9 [152[SAINSKYTL [9 FI-[4EGDKMPQLI [1 63APTQSFV 583 LPRHG1VGL 599 YGDNSAVTL 08 NAEVRYSIV (8201 GTITWI 899 DGNRVDL1 fl IPNKSLTTA [88 IGARIDREKL[8 11171 DEIFRLVKI[9 138 LFPATVNI[J 336 1PARAMVVN 380 IALITVK [9 F3 8 0 40 RPVFSNQFL 44 IPPLNQSAML [9 586 HG1 VGLT 1 601 DNSAVTSI 780 DSLFSWIV [9 814 LVMAVAGTI [1 9 LPLDNTFVA[9 [9961 YPVTEVP 171 NGVQNYELIE[7 347 DVNDNVPSI [7j (43J IAADAGKPPL [7 44] IDAGKPPLNQ [7( I4([VPPLTSNVr 7 [8J NLLLNFVI SSGTYIFA 16 139 FPATINIS I?98 DREEKDTV1 [232 TAILQVSVT1 00 Table XXXJIII-109P1D4 Table XXXIIII-109P1D4 Table MI111-11O91PD D v.1-B5101-9-mers 15i ars v.1B5101-9-mers Each peptide Is a Each peptide Is a Each peptide is a portion of SEQ ID NO: portion of SEQ ID NO: portion of SEQ ID NO: 3; each start position is 3; each start position is 3; each start position I, specified, the length of specified, the length of specified, the length ol peptide is 9 amino peptide is 9 amino peptide Is 9 amino acids, and the end adds, and the end acids, and the end position for each position for each position for each peptide is the start peptide is the start peptide is the start psition hius el t.ositio u e h position plus eight F3 1RAM6VNVT 1[e 18001 IADVSSPTS 15 [F742 TDLGU-HRVL F IPFRLRPVF 16 817] AVAGT I W M759 PD SLFSW I F 00 492 DADGPNAI F1oo311 VPVSVHTRP 15 1768 VNLFVNESV F 508 DAPPEFSLID 16 [301 IREEMPENV 1[i41 84] DVSDGNR 14 516 [DCRTGMLTV1 72 VPLIRIEED 14 897 OJSDGNRVTL 1A 520 GLTVKKL 16 83 1 EIFTTGARI 14 676 PPSNCSYE d61 156 ES16LPV 14 Table XiTV-1 09PI D4 744 LGLHRVLVK 16 161 PAAVDPDVG 14 v.1-A-10-mlers 791 EAPVTPNTE1H 162 F QNIFGLDVI 5nl Each peptide is a portion of SEQ ID NO: S EKDTYVMKV E 3; each start position is 999 TTEVPVSV 16 IPENAPVGT 14 specified, the length of DF E peptide is 10 amino MDSTI 15 276AIGENAKI acids, and the end 14F LACWFHSG 1I5 1 L MP 14 position for each peptide 34] MPEN GD 15 14 3 I w Is the start position plus 59 TAMQFKLWY 15 nine M.VNVTDV 1 67 YKTGDVPLI 15 [311 VNPVNDTVV LI23DREKLCAGI 15 [1 DE]LSENI1 148 1 P j ENIPLNTKI 14 58 AMQF28 46 15 43YLDYESTKEY 176 YELIKSQNI 151 (421 AAYDYEST 14 185 FGLDVIETP L(1I U426 YESTKEYAI 14~i 15271 KJDREKEDKY 198 F9i~ 1-0Q1 yGK 1981 MPQUVQKE 15 432 YAIKLLAAD4 261 NAPVGTSVE 1FJ 27LAADAGKPP 1141 1J DREKAESY 262 7APVGSTQ 1 465 VFTQSFVV I L29l qEDREEKOY 275 DADIGENA J 15 [HZ]_TQSFSI 350 6 2D6R Y [O j 313 EPLDREETP 15 493 ADSGPNAKI 14 _35-01__E3 3561 DIRYIVNPV L1 W509 APPEFSWC 14 67 E 360 IVNPVNDTV 15 1531 TILADNGV F53-91 14 673 _21 449 SAMLFIKVK 15 [541 LAKDNGVPP 14 7 E 517 CRTGV T 15 579] 0721VPENLPRHG 14 807 TSD KJLVA 532 KEDKYLFTI 11AVPVG 2 EKE7D M7FVS -T-1j 149 11 ETEISIPE 596 IPDPDYGDNSAY1 610 LENDFTIF[4 644 610 E4F5-66 644 ER [617 TIDSQTGV14 930 707 MNAEVRYSI1 666 VNDNKPVI 1[ 727 DQETGNITh [69 IAVDNDTGM 1 F-1 N 1 P 699 14K 00 Table XXXJV-109P1D4 v.1-Al-10-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine.

C-I [1239 1VITNNHV II 273FATDADIGENA 118 00 1345VDNkVS1 9FTKEYAIKUA 8 789STEAPVTPNT 8 [81DSDGNRVTLD 18 19 FSA KN 17 [1071 FYEVEVAILP 7 [385VDDDHG1 39 DHEIPFRL 7 1DEIPFLRP 17 797NTEIADVSSP 7 ~JTDLPIDLEE 17 IDLLKDLNL 6 44] LD LSLIP 6 17DVGINGVQNY 16 1 EGDKMPLIV 16 329ADGMPR1 514 SLDRTGMLT [I VTHNEYNFY 16 590 GITVDPDY 16 lIJADVSSPISDY I6 Table XXXV-IO0PI D4 v. 1-A0201-10-mers Each peptide Is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 10 amino acids, and the end position for each peptide is the start position plus nine I-I LLSG1YIFAV F29

SLFSVIVNL

00 00 Table XXV-0PD v.1-AJ 00 00 STable XXXVI-109P1D4 v.1-A0203-10-mers I Each peptide Is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position pus nine L 291 TREYAIL LLA ho] 14l AKPPLNQSA 10 14]1 IVKDENDNA jal 148j1 PGIQL11VSA 101 /4]1 QIKVSMDA 101 I490 MDADSGPNA 10 15001 K1!NLPDA 101 533 EDKYLILA i 59511PDYGDNSA 10 16361 QESYTFVKA 101 16481 GRVSRSSSA 10 l9jjlJ P~1VFVIA J101 ~1J 1ADNDTMNA ~71l VGGNTRDLFA 1ii01 744 LLHRVKA 10 771 LFNESXNA 10P E781 NLVRKSTEA 10 792 AErPNTEL6 10 807 TDYV1!LVA 10 '8231 TVVVIFITA 10 830 IAWRCRQA 10 83 RRQAPKA[0 846 KNKQ'EWA Wo 88: NEVTIEEKA 19271 TEKPDSPDLA 10j 933 PDLARHYKSA 10 938 HKSASPQPA 10 9651 E~LDNTVA 10i TI1 LLSGYIFAV [1] j 1YI5FA1 LAC ]9 IACWFHSGAQ W 5211 INKSLAM W~ 81 I TGEIFIGAR 1I~I 90:1 1 R!DREKCAG I] ijlJ H~FYEVVAI [91 Table XXXVI-1 09P1 D4 v.1-A0203-10-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide Is 10 amino acids, and the end position for each peptide is the start position plus nine 121LEOINNAP fl9 113 NDNAPLFPAT [E] 115NjSIPE~SAI 9 IJGEPQRSSTAI [9 I2IIVSIPENAP 9 ~JGISvrQHAT 119 F145VIQ AD '9 4]TADIGENAK jjFNLVSHIAR i9 J261 ARRLFHNAT I~l 13211 P1HKLLVLAS E[] 373N!PLNT 382LTVTDKDAD 425DYESTKEYAI lAIKLLADAG IW 21 P LNsSAM F49-11 DENeNAP IGIQLKYSAM E9 OLKVSNDAD 11 j491J MIDADSGENAK [IJ [501]1 IYLLGPDAP [I] F5961 DEDYGDSAV]I9 I6321 ESYFFYYKE][9 R911 GBVSRSSSAKI 16921 GIWVFQLW]I 1701 IVRNDTGMNAE 17171 GNTRDFAI II~I 17451( GkRVLAN I~ 771 I FNESVNAT I hi 1 EVRKSIEAP W 17931 FTT 1 E81 VKILVMVAG 91 00 00 Table XXXVI-1 09PI D4 v.1-A0203-1 0-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine

I

824 WVVIFTAV 9 311TAwRceQAP 9 8 NKQNSf-WAT 9 NTIEJMJA 9 98F! PDSPDLAR fI [jDLARHY1SAS 19 I~i1YSASPQPAF 19 L8J LDNTEVAC Table XXXVII-109PID4 v.1-A3-1 0-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 1 amino acids, and the end position for each peptide is the start position plus nine 743 DLfLH LVK 28 826 2IFIIWR 8 407 1RLPVESNQF 188 DV1ETPGDK 421 AAYLD STK[ [I1I AV '/VH F 4 1E SL2PN4LT2 F39 K7-912DK 4 817 \VBGTITVW 24 [17 (WHSAQEK 13 JELREKDTY23 832 2WRCEAPH 200 Q2VQ2LDR 298 RLEHLNATTG 5271 KQRE DKY 8101 22LYVAVA 813 IL Ali[22 Table XXXVII-09P10D4 v.1-A3-10-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acds, and the end position for each peptide is the start position plus nine [KJ DLLSPNK [1 F20- I 333 G j 435 KLLAqGKP 697 QV1AvprG 838 21QAPHLJ4QK 64 E2KLYKIDVP 73][ PL2RIEDTG 76 RIEEDTGEIF2 196 DKMPGVQK 136 IPVN T 478 11NNSPG~LTK o [487 KVLAM 5271CRIGlTVli( 20 523 ()TVK1REK J F6 I50 PP 2 65 1 RVSRSSSAKV 2 779 A20N~RK 1I CVWFHSAQE1 1115 LPEIFRLVK 1 163 AVDPDING 1 209 REEKDTYVKI19 [1 LLETAAlDY 1 I3J DKLF([11 590 GLTV1PDY9 617 TIRSQGVIR 19 623 GV!RP'~SFD 1 [673 FIRPPCSY 715J IVGNRDLF I [7 J (ThEKVD [9 [65 LVtXKTVPL 8 [218 KVWEGF P~ [01 HL~jAT{LTT(i~ *21 JLVJAS~GLM 1J 327VLSLM18 1GK Table XXXVII-109P1D4 v.1-A3-10-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 1 amino acids, and the end position for each peptide is the start position plus nine 464 1 PVETOQEVT18 504 1LLGPD&PEF 518 RTGMLTVKK J 624 VIPNjFDR 1 658 KV[INDVN 18 16741 IVEPSSYE L81 769 1NVNES 8t F8251 VVWIAW JMIMMKKK 18 [934 If DLRH ~SAS (2 42 J[ DLLKDLNLSL j7 (9 ][GERD~UCFY I 12111 RL.YKIELIE 7 [I6 I DVGINGVQNY[7 (308J LIIKEPLDR 17 1314 1 PLRE~PNH 1 SEIPFRLRP 17 43 I[UAA1DAG 503 JYLGP~PPE 17 1J~iI LW~LDR 1 [4J[GVPPLTSNVT 5821 NL1RH7IVGL F09IILDENDDFI 7j 635 KESYIEVK 17 693 V1 T 'JAVD IH 694 1 WEQVE1VDN [l7 65] WIVNLFVNE 803VSSPTSDYK 81 LV VBTr[7 001 Table XXXVII-109P1D4 j Table XXXVII-109P104 Table XXXVII-109P1D4 v.1-A3-10-mers v.1-A3-10-mers v.1-A3-10-mers c1 Each peptide is a portion Each peptide is a portion Each peptide is a portion of SEQ ID NO: 3; each of SEQ ID NO: 3; each of SEQ ID NO: 3; each start position Is specified, start position is specified, start position Is specified, the length of peptide is 10 the length of peptide Is 10 the length of peptide Is amino acids, and the end amino acids, and the end amino acids, and the end position for each peptide position for each peptide position for each peptide is the start position plus is the start position plus is the start position plus nine nine nine 870 1KIKHSPK 1727 FLIEDINFNA 48 QIMPEELNS E 156 SK RLA VD )15 1f42 TVINI IPEN 14 F37 NLIGDLLKD 16257 SIENAGTE1 1 -AINSKLPA 1 E- -PVG-lF153 1 00 E] RIDREKCAG 1 267 SVE15TDA 211 EKD E4KVK [if KLCAGBRDE 16J 276 AD15EKIH I23311 AILQVSVTDT I 1 i EVLPEIF 16 3151 EEENK (fs] [255 EVSIPEAPV 113 A D1FRL 16 324 FKL!YLA~DG El263 PVGTSVTQL AILVSM[DTh 16 341 354] SIDIRYIVNP 1 241 DT~IN~VFK [16J 344 N384 VIDHN1 1291 LVSNIRLF 16 1 347 DV J1DNSD 15 1 9) RVTCFDHEI14 34- 0M1 VNVTDV DE1YiVNPVN 1i E 491 363 PVD13SE 16 [1j VSENIPLNT 1 o KINYLLGPDA 375 16370 LSNIPNTK 15 549 [PLTSNVTVFVf 381 ALTVKDA 1I J 457 KDN0APV 15 1[PVETHNYNF 4161 FLLTMYLD 16 1514] SLDCRMLT 115 6 AV14SILDEN 423 YDYESTKEY [5591J[ SIEDQENSPJIS [E4 V LLADKPP [16 16261 RISEREK FsI I71O EVYS!GGN 455 16KVDENNAPI61 644 KADG~VSR 15 f7251 AIQETGNIT1 484 16QLVMDA 16671 PHIVPPSNC [fsJ fl74] GLHRVVKAN F52]KKLDR 16EDK 684fl LVPSIPGT 151 78011 TLNELVRKS 14 665 DVNHDNF7VF 1 16111 SYVNL 51 784 HELRKIEAP 685 VLSTEGTV 16 767 IVNLFVIESV 15 [7931 PVTPNTEIAD14 712 RYSIV~NTR [16) 859 P 799] E!DVPTS [F 7l22 EAIETG 16 862J 1RQIMKKKK 8231 TnVITA[ [748! RIvKAAfLG 16 H86] F&-3I-41KK [I 8VRRQAH LW 764 SwlFFVN 16950 I18ETE7SK9] QM K 785 j LVKS~EAPV L161) 11 i HI!QEN I)1sj 1879 KNUNFVT I 1 [812 KIVAGT [1 19651 ELEDI{FVA 15 NLLLNFVTIE [141 F8331 16 CRPHLF 16 j PVSVH]8PVG 15 [863fl H8rii [v] 902 RVILD1EDL J 16 ioi I PV~IVSNT [is5 [1 I V1D8RV 14 FS-2 ILEEIMGK 16 [12_ 1904 TLELPLEE 14 F96 svoc vr j1 f36 6ENIGDLK 14 906 DLeIlEQT FJ38 VLJGD(DL 15s [511 LENKSLrA 14 1967 PLDNTFACD 43 LLDLNLSLI 15 158 UAMqF1LW 14 TFVACDSISK [D KS5TTQFK 15 59 14TQF1VK [I4] #1V'ACD1SKC 118 E5ERLY1IIRF 515 124 KIRFEDIN 977 SI4KCSSS 00 00 Table X)O(Vi-1 09P1D4 v.1-A3-10-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 997 PVTTF 14PVS L(4 Table XXXVIII-109P14 v.1-A26-10-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 167 32DVGINGVQNY 319 ETPNKLL 1 iii EVALPDEIF j 118 EIFRLVKIRF27 88 DVIETPEGDK25 [i1 vRYSIVGGN25 109 4EVEAILPDE 350 DNVPSIDIRY24 367 4LSENIPL 740 DVTLGLHRV24 820 GT2TW 1F 4 277 DIGENAKIHF 428 STKEYAIKLL 23 71 DVPLIRIEED 130 EDINDNAPLFf: 403 EIPFRLRPVF( SPVFTHNEYNF 22 7jJ ETGNITLMJEK 2 F7D ET91 22 FL0 E22REEKDTY 427 ESTKEYAIKL21 i DNSAVSIL 21 928 2TFKPDSPDL Table XXXVIII-109P1D4 V.1-A26-10-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 58 I 1TAMQFKLj 191 ETPEGDKMPQ2 213 DTFVKVKVE 20 255 EVSIPENAPVI2 3j[fDVNDNVPSID (2 366 DWLSENIP 20 494 DSGPNAKINY 20 [555 TSIIDQN (673 FVPPSNCSY (737 EKCDvDLGL 26 776 VTNATlNELj) 2] RV2LDLPIDL 0 F TTF PVSVH 2 12JEVPVSVHTRP o TVINISIPEN o [2511 ETEIEVSIPE 19 316 DREETPNHKL 19 I GVIRPNISFD 19 2 D1DNKPVFI9 (9((TFQV1AVD 17641 SVVVNLN [9 [8p2 DVSSPTSDYVjW [824 VVVIFITAV I~ V(19 [42 1 DLLKDLNLSL1 6LWKGDVPL18 DTGIFTGA 18 91( EITTGARID 1 2T j LVSNIARRLF Ki r91 Y~s 18 4611 DNAPVFTQSF 18 574 EYNFYVPENL 59DYGDNSAVL 8 692 WFQVIAV8 7 IVGGNTRDLF 8] Table XXXVIII-19P1D4 v.1-A26-1D-mers Each peptide Is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is amino acids, and the end position far each peptide is the start position plus nine 761 SLFSWIVNL E [833 3WRCRQAPHL E 953 ETPSKHHI18 33 lEMPENVLIGD 17 113 AILPDEIFRL 17 1178 I UKSQNIFGL L71 (241 DTNDNHPVFKI17 262 APVGTSVTQL 7 2931 SNEIRFHL 363 PVNDTVVLSEI1 554 VVFVSIIDQ 17 j632jI DREKQESYF 1 [i7] SVTNATUNEI 809 DYVKILVAV 1 823 1VVVIFTA 17 f1 S 16 9fl EEEMPENIG NVLIGDW(D 16 VGDLLKDL 16 117 DEIFRLVKIR 16 172 GVQNYELIKS [ij 3 ITIKEPUJRE L31[ FTDHEIPFRL 6 10 gP6SNQFE :5:2LT2KKLDRE J29DREKEDKYF ~6 53 EKEDKYLFTI1 J12( ENDDFTIDSQ M 662J

R

741 VDLGL6RV 750 LVKANDLGQP1 7991EIADVSSPTS 80 ]ADVSSPTSDY 82: I1WIFIT 00 FTable XXVIII-109P1D4 v.1-A2-&1D-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide Is the start position plus nine 972 FVACDSISKC WIj 972 S 1:0D06 n16I) Table XXXIX-1 09P1 D4 v.1-0702- 10-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 10 amino acids, and the end position for each peptide Is the start position plus nine 547 VPPLTSNVTV18 596DPDYGDNSAV18 676 PPSNCSYELV 18 [R ITPNPENRQMI 8 945 QPAFQIQPET18 1003 VPVSVHTRPV 18 139 FPATVINISI 17 VPENLPRHGT 17 877 SPKNLLNF 17 72 VPRIEEDT 1 444 PPLNSAMLF 510 PPEFSLDCRT 858 907 PIDLEEQTM16 954 TPLNSKHHII 115 LPDEIFRLVK 15 136 APLFPA5IN 335 EARA 15MVN W 532 KEEK5YLTIL 817 AVAGTI 1 8961 4 LSG1YIFAVL1 IGDLLXDL 14 J1 LWKTGDVPL 1 119 IFRLVKIRFL 1291 IEDINDNAPL [4 319 IETPNHKLLL 1 3 VNPVNDTL I IPFRLRPVS 14 [898 1SDGNRVT 4L 947 AFQIQPETPL14 959 KHHIIQE 14 966 LPLD4FVAC 1 42] D13KDLSL 1001 IPRDEHC 3E J 113 AILPDEIFRL1 Table XX)IX-109P1D4 v.1-B0702-110-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is amino acids, and the end position for each peptide is the start position plus nine 16 70LPAAVDPDVG F3 2821AKHFSFSNL 113 31]3 EPLDREETPN1l1 r3331 3 [62 J1NPVNDTWLS [4Z1[LAADAGKPPL j3 r4 i SPGIQLTKVS I1 15.!1ILAKDNGVPPL 13 58221NLPRHGTVGL 1 j59 JDYGDNSAVnL 13 1601 DNSAVTLSIL 067j PSNCSYEL 13 714 SIVGGNTRDL 13 735 LMEKCDVTDL13 [5][EKCEVT3LGL IANDLGQPDSL j3 83 13WRCRQAPHL13 874 K3]SPKNL 929 KPDSPDLARH Table

XL-

109P1 04 v.1-BOBlO-mers No Results Found.

Table

XLI-

109P1D4 v.1- 81510- No Results 00 Found.

Table

XLII-

109PID4 v.1- B2705- No Results Found.

Table 109P1 v.1- 82709- 1-mers No Results Found.

Table XUV-109P1 D4 v.1-84402-i 0-ers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide Is the start position plus nine 3171 REETPNHKLL 476 PENNSPGIQL 532 KEDKLTIL[ 912E 176 YEUKSQNIF 773 NESVTATL I 1PENVUGOLL 82 1 21FTGR j2J IEDINDNAPL J 1491 PENSAINSKY 21 1 PEGDKMPQLI21 [1 REEMPENV 2 98 AGIPRD CF 20 [IhJ AILPDEIFRL [20] 27 GENAKIFSF 371 SENIPLNTK 120 Table XLIV-1 09P1 D4 v.1-84402-i 0-mrs Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine 633 REKQESYTY 110 VEVAILPOEI 3I 1 EEMPENVLIG 118 [781 EEDTGEIF~F8 130 EDINONAPIF 402 HEIPFRLRPV 70][AEVRYSIVGG 18 L3i VUGDLLKDL 282 AKIHFSFSNL 318 EETPNHKV1 9 ETPNHKLLVL 17 P41 17ELETAA 42 STKEYIU 7 49 SGPNAKINYL J761 ]jSLFSWIVNL jj 11171 DEIFRLVKIR Ii i1hJ EIFRLVKIRF [252 ifTEIEVSPEN [6 if2 APVGTSVTQL je [333J GLMPARAMVL1 [3Z7I NIPLNTKIAL 5191 TGMLVKKL 6] 16451 AEDGGRVSRS 1 753 ANDLGQPDSL 1 790 TEAPTPNTE 1 820 GTIVVVIF 1 [9301 PSPOLARHY 16 riiFEVPVSV1-TR 1 24 11QEKNTIREE 1 [8 NLSLIPNKSL [s L5i NKSL1TAMQF 1 119 IFRLVKIRFL 1 123 VKIRZFLIEDI 5 137 PLFPATINI 15 1 IETPEGOKMP 1 205 KELDREEKDT t Table XLIV-I 09P1 D4 v.1-B4402-10-mers Each peptide Is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is amino acids, and the end position for each peptide is the start position plus nine 206 ELDREEKDTY Ls F-0]EEKDTYVMKV'[s LVSNIARRLF 1 12931 SNIARRLFHL 1 390ADHNGRVTC

F

403 EIPFRLRPVF1 407 RLRPSNQF 7II ESTKEYAIKL [43-0K1YALLAA [96VDSDGNRVL 941 SASPQPAFQI1 92 L5f SGTYIFAVL1 119 I FHSGAQEKNY I4 LEiJ MPENIGDL14 108 YEVEVILPD 1 [312 KEPLDREETP O350NVPSIDIRY] 351 NVPSIDIRYI1 3 if IPLNTKIAI 397 j[TCFTDHEIPF[1 I2IIYDYESTKEY E1 4571KDENDNAPVF[ 14 DNAPVFTSF I 494 DSGPNAKINY [04 LGPDAPPEF II [51][PEFSLDCRTG 4 [S2i]KLDREKEDKY 4 [4J PPLTSNwVTF 590 GIVPDY 14 598 DYGDNSAV 14 LSENDDF 14 61 FTIDSQTGV1 14 00 00 Table XLIV-1 09P1 D4 v.1-B4402-10-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 1531] EKEDKYIFTI] [5661 NSPVFTHNEYI 568 PVFTHNEYNF 131 573 NEYNFYVPEN 1 [5741 EYNF

PENL

[ii f DENDOFTIDS f3 SFDREKQESY6301 636 QESYTA 673 FIVPPSNCSY13 715 IVGGNTRDLF13 724 FAIDQETGNI [a 728 QETGNITLgE 3 747 HRVLVKANDL 13 I 9] TEIADVSSPT[3 04 SSPTSDI 1 F8s73

I

8 KK SPKNL 13 876 HSPKNLLLNF 18891 EETKADDVDS1 19021 RVTLDLPIDL 939 YKSASPQPAF 947 AFQIPETPL1 [953] EPSKHHI 9 J[ IQELPLNTF[3 I2 1 DW(DLNLSL 12 Wf 1TAMQFKL2 KTGDVPURI12 I1 IRIEEDTGEI 12 Ir IEEDTGEIFT 12 IK REKLCAGIPR 1i9j1j GIPRDEHCFY 12 [1111 EVAILPDEIF[2 145 NISIPENSAI [151 NSAINSKYTL 2 11921IITPEGDKPQL Table XLIV-109P1D4 v.1 -84402-10-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is amino acids, and the end position for each peptide is the start position plus nine F26 FPQRSSTAIL If1 241 TDTNDNHP VF I2 250 KETEIEVSIP 12 299 LFHLNIATTGL 300 FHLA1TGLI 12 302 LNATT[TI 1 I316IDREETPNHKL j2 Isr TWLSENIPL [12 F3-671 12 399 FTDHEIPFRL 12 1417]1 ILETMYLDY j]j [-4-2611 ETFAK 1426 IYESTKEYAIK Iz 528]1EEDn LDREKEDKYL 1i] [I54IILAKDNGPPL 12 [~jj IDQNONSPVF 12 SO I PENLPRHG1V 12 1601) DNSAVTLSIL 12 652 I 1 []VDVNDNKPVFf PSNCSYELVL 6 90 1J NPG1VFQVI f~ I GGNTRDLFAI ff2] 7261 736 MEKCDVTDLG 7831 NVRKSTEA 56 TPNPENRQMI2 888 IE[ADDVD 2 9112LEEQTMGKYN 9191

E[

9261 2 9591 KHHIIQELPL 980 KCSSSSSDPY Table

IXLV-

109P101 U v.11- B5101 00 flNo IiResults I Table XLVI -109P1D4v.1-DRB1 0101- Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is amino acids, and the end position for each peptide Is the start position plus fourteen 8081 S D YV KI -VM--VAGT 11 ~6 F- J[ 1YIFAVLLACWFHS 134j SFl-1 GTSVTQLHATDADIG 134] F4-J21 GIQLTKvsAmDAS 5-3 49fJj NAKINYLLGPAE F33 F281 HFSFSNLVSNIARRL j 173J1 VQNYELIKSQNFL~ [405fj PFRLRPVFSNQFLLE J [117]1 DEIFRLVKIRFLIED j28 iT55]1 NSKYTLPMAVDPDVG] 28 F2--1 RRLFHLNA GF~I28 F7Zi-0]1 EVRYSIVGGNTRDLF] 1-8 F797]1 NTEIAVSSPTSDYV 28 F8--21 LLNFvTIETAV F28 F9-]51 QPAFQIQPETPLNSK IN~ Fl-09]1 EVEVAILPDEIF=RLV 27 [~F11SNQFEMDY F27 TSDYVKILVAVG n27 F95-0if RIDREKLCAGIPRDE] F105][ HCFYEVEVAILPE F26 Fl-4-111 ATYINISIPENSAIMN]2 I 8711 LDVIETPEGDKMPQL 26 F2-JI FSNLVSNIAR [2F 6 KEYAIKLLAADAGKP 1[26 F4fjEYAIKLLAADAGKPP iI1 F5--81 F11LAKDNGVPT 26 [S-721HNEYNFYPNL PRH 26j [59-61 DPDYGDNSAVrLSIL 126 73f] KCDVTDLGLHRVLV-K 26 823] TVVWTAWRCR 26 831 JTAVVRCRQAPHLM2 :733:] EMPENVLIGDTLKL 2 Table XLVI -1O09P1 D4v. 1-DRB1 I 0101- 15-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of pepffde is amino acids, and the end position for each peptide is the start position plus fourteen [41 GDLLKDLNLS-LJP-NK [25 62 QFKLVYKTGDVPLIR 251 [l-D-41 EHCFYEEALD 251 [176j YELl KSQNIF gL 5 [216J VMKVKVEDGGFPQRS 251 [2-2-31 DGGFPQRSSTALQV I~ flji ARRLFHLN1GTII F51 LIVLASD)GGL-MPARA 25~ I3-3ARAMWVLNTVD F2 I II AIKLLAADAGKPPLN-f5 (453-4j IKLLMADAGKPPLNQl 25~ 58011 PENLPRHG1V-G-LITV1 E25 FE II NDDFTIDSQTGVIR E25 16401 JTFYVKAEDGGRVR 25~ J TGNITLMEKcprDTL l F~~jSWIVNLFVNESvrN f~I [~i[VKILVAAVAGfTI[TWI 251 [~[PTTFKPDSPDLARY 25 F93-][ ARHYKSASPQPAFQI n25 [EEi NYTIREEMPENU 24 fDLNLSLIPNKSLT 24 7 jLIRIEEDTGEIFTTG 24 11611 POEIFRLVKIRFLiE 2 114511 NISIPENSAINY 24 13-2211 NHKLLVLASDGGLMP 124 [32]4 KILLVLASDGG LMPAR12 ]ASDGLMPARAIVILVI 24 [3 I ][DGGLMPRMLN [24 jRYIVNPVNDTWLSE I2 f 1VSIPENNSPGIQLr [24] f]j NNSPGIQLTKSM [24 F4881 [VSAMDADSGPNAKI-N 49]1 A(INYLLGPAPE 2 :5 HGTVGLI1VTD PDYG R EDO TINWDVNDNKPVFI 14 7J KPVFIVPPSNCSYEL :698 VLWDNDTGMNAEVR 24 Table XLV 1-1O9PID4v.1-DRB1 0101- Each peptide Is a portion of SEQ ID NO* 3; each start position Is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the Lstart Position plus fourteen R72 YS IVGG NTRDLFAJ- F24 fl G LH RVL VKAN DL GQP 214 F70[DSLFSWI/NLFVNE 1[24 [ff I1VWVFITAWRC r114 SF1 FvTIEETKDVD [24 F9-5if GNRVTWDLPIDLEEQ1 E24 Fq J[]YNwvTTPTTFKPDSP 975j[ CDSISKCSSSSSP F24 1 I LLSGTYIFAVLLACV I 3 [i i H KDLNLSLIPNKSLiT g3 [7Wflj EEDTGEITGAI i [2911 IEDINDNAPLFPATVl 151 1 NSAINSKLPV 23 [i z11 DVGINGVQNYELIKS 1231 [2-8i1 NAKIHFSFSNLVN I11 SNLVSNIARRLF2131 [34211 LVNvTDVNDNVPSID 231 [349][ NDNVPSIDIRYVNP j231 [37011 LSENIPJKAI 231 S379 1f KIALITDKDADHNl n23 531 i EKEDKYLFTIL-AI(DN1 23 I f5341 DKYLFTILAKDNGVP E231 ifvppLTsNvrVFvsii &23 I 50if SFDREKQESYFV 213 fli GGRVSRiSAKVT1N g13 F6-6-3I WDVNDNKPV F 313 66911 NKPVFIVPPSCE n23! [691 NCSYELVLPSNT 213 F6-8-01 CSYELVLPSTN231V 11j [Z ii1 INELVRKSTEPT 1131 [If KILVMAVAG1VV 23 jf] AGTITV'wiFTrAv f 3 TIVFITAWR gI [82-41 WWIFTVRR I23 Lj]QKNKQNSEWATPN1 23 [916 MGKYNW TPFK 23 r 963 IQELPLDNTFVACDS M2 00 00 Table XLVI -1O9P1D4v.1-DRB1 0101- 15-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is amino acids, and the end posilion fr each pepfide Is the start position plus fourteen [II GTYFAVLLACVVFH f [1-2611 RFLIEDINDNALPI INDNAPLFPATVNI[2 LIKSQNIFGLDVIET 1 [2511[ ETEIEVSIPENAPVG f2 132811LASDGGLMPARAMVL 40211 HEIPFRLRPVFSNQF f] F44-2j GKPPLNQSMFK j F46-21 NAPVFTQSFV'IVSIP T j[LTKVSMDADSGPNA 5021 NYLLGPDAPPEFSLD 122 5101 PPEFSLDCRTGM LTV 2 535 1KYLFTILAKDNGV r2 2 544][ DNGVPPLTSNvrvFvI 22 557Q FVSIIDQNDNSPVFT 22 615] DF11DSQTGVIRPNI [22 IF683ELVLPSTNPGTWFQ 22 F69-2I1 GTWFQVIAV-DNDTG 22 75311 ANDLGQPDSLFS-WI 22 F7-5-61 LGQPDSLFSWIVNL [i5-9[ PDSLFSWVLVNf [800][ IADVSSPTSDYVKIL 87151[ VMVAGTI1VVVVIF IE2 9[391 YKSASPQPAFQIQPE I2- 79471[ AFQIQPETPLNSKHH L 2 10011 FEVPVSVHTVi Q [2 1NwAIQwtrL V P [E 10811 YEVEVAILPDEIFRL [21 18411 IFGLDVIETPEG-DKM [i211 3611 PVNDTVVLSENIPL-N1 E21 54111[ LAKDNGVPPLTSNVT I1H 7221f DLFAIDQETGNITt.M []i 1431 VINISIPENSNS [20] 2175[YVMKVKVEDGGFPQRI E2O 22]2[ EDGGFPQRSSAL r-O 24611 HPVFKETEIEVSP 2:fjEIEVSIPENAPVGTS2 ITable XLVI -109P1D4v.1-DRB1 1- 0101- 15-mers Each peptide is a portion of SEQ ID NO: 3; each start position is Specified, the length of peptide Is 15 amino adds, and the end position for each peptide is the start position plus fourteen- 3231 HKLLVLASDGGLMPA p f-3-61 TDVNONVPSIDIRYI 110 F4-1 -IEtKEYILAA F2i F45911 ENDNAPV FTQSFVTV 110 46-311 APVFTQSFS-IPElol F47-0I1 FVTVSIPENNSPI Nl [522J LTVVIKKLDE KEDKY 20 r 619]1 DSQTGVIRPNISFDR 20 [7if][ VNLFVNESV~ L 20 78311 NELVRKSTE2PVT1 883J[ LNFVTIEETKADDVD2 944][ PQPAFQIQPETLS2 9-921[SDcGYPVTTFE-vp-v-slI W1j FKLVYKTGDVPLIRI jig F641 KLWYKTGDVPLRIE 19j~ Ejj LVKIRFUIEDIN-DNA1 19 E82j QNIFGLDVIETPEGD] [19 M3061 TGLITIKEPLDREET f19 J352f VPSIDIRYIVNPVND I19 36-511 NDTWVLSENIPL1NT~ 19 42-011 TAAYLDYESTKEY-AI 19 F6-001 KNYLLGPDAPPE-FS] 19 6-541[ AmuSLENDT 19 F6-961FQVAVDNTGNE1 [i ITLMEKCDVTDLG-LH 19 F[8T] YIFAVLLACWFS IF-1 DK~l JLACWFHSGAQEKN-Y 18 4011 IGDLLKDLNSLP 8 [soj[ SLIPNKSLTM F I8 :rj41NKSLTTAMFK-LWVK1 [18 TGEIFTGARIDR--EK Ha 13311 NDNAPLFPA1II 181 13611 APLFPATV1nIS F18 E70]1 INGVQNYELIK-SQNI- jI 245fl NHPVFKETEIEVI 1 25711 SIPENAPVGrsvrQL 1 293] SNIARRLHLNATTG i ITable XLVI -109P1D4v.1-DRBI 0101- Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen [3191 ETPNHILLVLAS-DGG [a- [411 VFSNQFLLETAL 5-8 [423j YLDYESTKEYIL Rf [4560]1 AMLFIKVKDENDNP [a8 [aflj FYVKAEDGGRVSRSS f8 [Ell GGNTRDLFAIDQETG [a- LVKANDLGQPDSLSS [E8 [7 1[l LFSWIVNLFNS [a :7 61 WIVNLFVNESVTrNA L81 :7::81 NATUINELVRKSE F187 779f ATUINELVRKSTA F18 8701 KKKKKKHSPKLN Ffa8 :9181l KYNWVUTPTTFKPDSl :NfIISDPYSVSDCGYP18[a 99311 DCGYPVTTFEVPS gf :9:9]5 GYPVITFEVPVSVHT ff8 Table XLVII -109121 D4v.1-1 DRBI 0301 Each peptide Is a portion of SEQ ID NO:, 3; each start position Is specified, the length of peptide Is 15 amino acids, and the end position for each peptide is the start position plus fourteen

IGDLLKDLNLSLP

EVPJLPIEIFLV-K1- 3~2 [gi GNRvTLDLPID-LE-EQ1 31i [flj ENVLIGDLLKLS El LIRIEEDTGEIFMTG 29 Z ICAGIPRD)EHCF-ENVE 29J A12 IRFLIEDINDNAPiLF 29 NYLLGPDAPPEUFSLD j I 89 3j ADDVDSDGNRTL 281 65]1 NDTVVLSENIPLT 7 60:5] VTLSILDENDD IDf[ 67]i PVFIVPPSNCSYELV J 00 00 Table XLViI -1O9PID4v.1- DRBI 0301 15-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 15 amino acids, and the end posifion for each peptide Is the start position plus fourteen Fr904 TLLIDEQMGK 14j1 DLNLSUPNKLTAl§ I1 NKSLTTAMQFKLWYK fg F3-7]1 SENIPLNTKIAUTV fg 525 VKL K YLIFT Ig [6-113 NSDDF11DSQITGVI 2 RPNISFDREKQEr g16 20j1 QKELDREEKDTYVMK 115 2-75 DADIGENAI<HFSFS [s [2891 SNLVSNLARRLFL 21 DHEIPFRLRPVFSNQ N 5FS-j1 PPEFSLDCRTGML1V g~ F5-6]6 NSPVFTHNEYNFYVP R251 66 2 NWDVNDNKPVFIVP 25 [7-1-31 YSIVGGNTRDLFAI1D 1251 1161 PDEIFRLVKIRFLIE-112-4 171DVGINGVQNYEL1KSj E24I [3951 RVTCFTDHEIP=FRLR 2~ 7211 RDLFAIDQETGNIT ]g~ 32511 LLVLASDGGLMPARA 231 f628 NISFDREKQESYrFY 231 [945 QPAFQIQPEPN( E23j 1161 PMAVDPDVGINGQ W8J1VSAMDADSGPNAKIN 22 1925 1PTFKPDSPDaRHY 9701 NTFVAODSISCS n22 [1i6]5 DPDVGINGVQNY-ELI 211 323 HKLLVLASDGGLMPA 21 4075 PFRLRPVFSNQFLLE 2-1 5381 FTILAKDNGVPPL-TS1 21! 698]1 VIAVDNDTGMNAV 21 7591 PDSLFSWIVNLFVNl21 96j IELPLDNFAD i 6311 FKLWKTGDVPUR Jo 1281 LIEDINDNAPLFPT2 1761 YELIKSQNIFG-LD-VI 20 Table XLVII -1O9P1D4v.1- DRBI 0301 15-mers Each peptide is a portion of SEQ ID NO: 3; each start Position Is specified, the length of peplide is 15 amino acids, and the end position for each peptide is the start position plus fourteen 288 FSNLVSNIARRLHL [20 14131 SNQFLLETAALY [210 P341 IKLLAADAGKPPLNQ [20 1580 PENLPRHG1VGLITV2 F691 FQVLAVDNDTGMNAEl F2 r8013 VSSPTSDYVKILVMA 120 r8-6] NRQMIMMKKKKKKKK 120 r9-81 PIDLEEQTMGK-YNWV 20I 928 IFKPDSPLRYS 201 R1 EHCFYEVEVAILPDE ]N f101EVEVAILPDEFR LV Hf~ E11 DEIFRLVKIR FLIED [182 QNIFGLDVIETG nij [186 GLDVIETPEGDKMPQ W 1901 IETPEGDKMPQLIQ 9 F811 MPQLIVQKELDREEK 19 E32]1 svrDTNDNHPVFKE-T]I91 E305] TTGLITIIKEPL-DREE1 N9 331j D G G LMPARAM VLNV 19 15 QFLLETMYLDES j9 421 MAYLDYESTKE YAIK 19~ 52 LFIKVKDENDNAP*VFH 518 1RTGMLWKRE [19 19 TGMLTWKKLDREKE I~9 567 SPVTHNEYNFfWPE 19 5188 1VGLITVTDPD D [9 6-82 YELVLPSTNP-GTVVF R9 J RYSIVGGNTDLFAI1 ff1 7301 TGNITLMEKCDD 9f 1746LHRVLVKANDL-GQ-D] 19~ [79 EAPVTPNTEIADVS f9! 1jTAWRORQAPHLKM- i19 IgJAPHLKMAQKNKQNSE 19 Ig RQMIMMKKKKKH D191 gIj MIMMKKKKKKS 1911 Table XLVIII 109P1 D4v.1- DRBI 0401-15-mer's Each peptide is a portion of SEQ 10 N0. 3; each start position Is specified, the length of peptide Is 15 amino acids, and the end position for each peptide is the start position plus fourteen 1i7'311 VQNYELIKSQNIFGL]11 [2-8-51 HFSFSNLVSNLARL F28! f-6-j01 PPEFSLDCRT-GMLTV ~J 651311 NDOFTIDSQTGVIRP 281 40-I1 IGDLLKDLNLS-LIPN [16 iF4-6I1 DLNLSLIPNKSLTFA 126 5][NKSL1TAMQ F 2LW [61 IRFLIEDINDNAPLF _[26 DVGINGVQNYELIKS- 216 fJ[ SF0 IRYIVNPVNT [1 I sw fl DNGVPPLTsNVT F21 555 i TVFVSIIDQNDNSPVl 26! 170411 DTGMNAEVRYSIG [g F7 6flj WIVNLFVNESVTN 126! F7--9j ATLINELVRKSTA 26 [797J( NTEIADVSSPTSDYV 26N 821VVVVIFITAWvRC-R ]g r8-271 VIFITAWVRCRQAH 116 F8931 [ADDVDSDGN-RVLDL fN W13] IQELPLDN-TVACDS [16 Ei1 1YIFAVLLACVVFH-S-2 Ifl ]CVVFHSGAQEKNYRTI J 11I EHCFYEVEVAILPDE 1122 I I DEIFRLVKIRFLIED F2 FI2 4Q KIRFUEDINDNAPL f22 [16fII RRLFILNATTGT F4 E Il SNQFLTM DE F22 [EJfl TQSFVrvsiPENN-SP j F6-1]1 NISFDREKOESYF 2~ 767I0[ KPVFIVPPSNSE R22 17fiNCSYELVLPSTWP-GT 22 RDLFAIDQENT 22 768]l VNLFVNES-VTNATLI 071] TSDYVKILVAAVAGT JN22 882I LLNFvrIEETAD 122 918 KNwvrTTPTFKPOS fE 00 00 ITable XILVIII 109P1D4v.1- DRB1 0401-15-mets Each peptidle Is a portion of SEQ ID NO, 3; each start position is specifed, the length of peptide Is amino acids, and the end position for each peptide is the start positionlusforte 92-51 PTTFKPDSPDLARHY [F2- [9-]61ARHYKsAsPQPAFQiI g [96]1 DNTFVACO)SIS-KCSS 9 6 r G1YIFAVLLACWFH IFo 27 I NYTIREEMPENVULG 1o 36I ENVUIGDLLKDLNLS 11o F3-71 NVLIGDLLKDNS 9o 41]1 GDLLKDLNLSLIPNK] 20 18] NLSUIPNKSLTTAMQ 12E 97 1 CAGIPRDEHCFYE F11111f EVPJLPDEFLK 2~ IU l1I VAILPDEIFRLVKIR 12R1 1:I22flI LVKIRFLIEDINDNA] F20 135j1 NAPLFPATVISIP 2 101PATYINISIPENSAI 20N [14I[f VINISIPENSIS 201 KYmPAAVPDVGIN F20 [i-8-1if SQNIFGLDVIETPEG 0 FI184411 IFGLDVETIDEGDKM 1 231]1 I STAILQVSTTD 232]1 TAILQvsvTrDTNDN-H 26 23411] ILQVSVTDTNDNHPV E2I [245][ NHPVFKETEIEVSIP E20 [253][ E iEVIEAVT 26 1GTVLHTADIGE~ 281][ NAJFFNVSNI 20O F289 rSNVNIRLHLN 120 F3121 KEPILDIREETPNHKaLL E F32-21 NHKLLVLASDGGLMP 120 F3--31 HKLLVLASDGGL-MPA 20 3-31 ]DGGLMPARAMVV 20 FJ]ARAmvLvNVTDVNDN gO: 331 RAMVLVNVDVNDNV 20 34911 NDNVPSIDIRYIVNP 120 35711 IRYIVNPVND1WLS N~ 35811 RYIVNPVNDTWLSE 20 ITable XLVIII 109PI D4v. I- :7DRB10401-15-mers Each pepide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen F3651NDTVVLSENIPLTK 1E2 j DTVVLSENIPLNTKI110E [377j[ NTKIALIVTKA 20E 37-9]1 KIALTVTDKDADHN @10 33 I[ NGRVTOFTDHEIPFR1o 1IPFRLRVFSNQFLLE [42i II MYLDYESTKEYAIK 110 [47j[ TVSIPENNSPL 201 F82 GIQLTKVSAMDADSG JFoi W1 [VSAMDADSPNA<IN20 NAKINYIIGPDAPE 110 Ef LTWKKLDREKK 1101 [53411 DKYLF-LAKDNV 20 154flj VPPLTSNVT FVSI I F11 TSNVTVVSDN [2-0 558]1 VSJIIDQNDN SPVFH10 58 1f PENLPRHGTVGLITV I [6 fj[ TISILDENDDFT IDS i E01[TFYVKAEDGGR R [20 F6--81 GGRVSRSSAKVTN] [20 F658] KvTINWDVNDNP [E) F6-611 INWDVNDNKPVFIV 20 F6-J21 YELVLPSTNPG1V 2 69211 G1WFQVIAVNT 16951 VFQVIAVDNDT-GMN-A] 29b [698j VIAVDNDTGMAV 9~ [712][ RYSIVGGNTRLA 92 F7-I0[ TIRIDLFAIDQEGI 20J [2Q] LFAIDQETGNTE F20J 73 8KcDvTIDLGILHRV 110 431 [DLGLHRVLVKNG 210 147 [HRVLVKANDLQD 210 75 ][ANDLGQPDSLSI0 75I9]PDSLFSVIVNL"-FVNjo li2:]LFSWIVNLFVNESV j 0] :7jjSWIVNLFVNESVTN 110]1 [Table XLVIII 109P1D4v.1- DRBI 0401-15-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide Is 15 amino acids, and the end position for each peptide is the start position plus fourteen 176711 IVNILFVNESVTNATIL 176911 NILFVNESVALI E2O] 177811 NATUINEILVRIKSTEAjf o r80011 IADVSSPTSDYVKIL Io] F881SDYVKILVAGTI o 810]1 YVKILVAAVAG11TV fo F8111VKILVMAVAGT1TVVjr2o] [812][ KILVMVAGTITWV f2o] [Efl VMAVAGTI1VVWIF 1[201 1819 i AGTI1VVIFITAV[o 82 lTIWVVIFITAVVR 110! 182lI WTVVIFITAWRC 110! F839IAPHLKAQKNKQNSE] F8-791 KL±NFvnEEA 110! F86[NLLLNFvTIEETKAD LNFvTiEETKADDVD 900][ GNRVTLDLPILE 1101 F9][l TU)LPIDLEEQTMGK 110 F906i1 I 94711[ AFQIQPETI.NSK-II 196011 H-HIIQELPLIJNTFVA [~CDSISKCSSSSSDPYJj FI9i1 GYPVTTIFEVSHT1o [11l VLLACWFHSAE ni8! LLACWFHSGAEN1 1911[ FI-SGAQEKNYTIREE[1 [i51J1 LIPNKSLTMF. [ni8 773]1 PLIRIEEDTGEIFT 18 :8]EEDTGEIFTTGARID] 18 :8:5]1Fi-GARIDR A 81 :13]1 AILPDEIFRLVKIRF 1 13711_ PILFPATVINISIPEN 18 44][ INISIPENSAINSKY [181 1481j IPENSAINSKYLAj8 196] DKMPQUVL DR 201l LIVQKELDREEK Y18 00 00 ITable XLVIII 109P1 D4v.1- DRB1 O401-15-mers Each peptide is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is amino acids, and the end position for each peptide is the star position plus fourteen [2201 K(VEDGGQ-RSTAI [8 [.28Q QRSSTAILQVSVTDT 18 F25f1 IPENAPVGTsvTQLH [iaj I 2621 APGST ATDA I8] [28211 AKIHFSFSNLVSNIA [18E 2931 SNIARRLFHLNArG fjI] 298][j RLFH-LNATTGLMK] 18 I~l ITIKEPLDREETPNH] [1 VL NVTVD-S IRa F346ff TDVNDNVPSIDIRY-I 18 F3-50r1DNVPSIDIRYIVNPVI1i-8 36-311 PVNDTVVLSE-NIPLN 118 F3-611 LSENIPLNTkIALIT 118 F38 1vDKDADHNGRvTCF [a 406 FRLRPVSQLT Hf8 44011 DAGKPPLNQAMF 18 452 LFIKVKDENDNAPVF 18 460 NDNAPvFTQsFvrvs E18 46 PvFTQSVrV-S-IPEN 18 P-8fJ KVSAMDADSGPNAKI 1118R F53-11 EKEDYLFTILAKDN E18~ W1j VFVSIIDQNDNSP-vF Jjl-8 5-6-8] WV~NY EN 18 n] 58PNLRGVG a F551TDPDYGDNSAVTLSI Eat ~598[ DYGDNSAVTLSILDE [18 609] ILDENDDFTIDSQTG [18 F6-1-8j IDSQTGVIRPNISFD [R8 F6251 IRPNISFDREKQESY 118 I 45]AEDGGRVSRSSKV 18 16591 VTINWDVNDNKPVF] 18 F6891 TNPGFVAD [a F74-1DvTDLGLVKA [a] S F18a L G-QPDFSWVLE '761]1_ SLFSWVNV NE 1 I 7701 LFVNESVTNTU NE [18 Table XLVIII 4-1IO9P1D4.1- DRE31 0401-15-mersJ Each peptide Is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide Is the start siti on Ius fourteen fffl1 SVNATLINELVRKS IE18 79-61 PNTEIAD)VSSpTSY f E813 ILVMAVAGTITVWV 18 F83-I W RC RQAP TH LKMQK [iQa5:3:] QAPHLKAAQKN-KQN-S [I WATPNPENRQMIMMK 181 [876 1HSPKNLLLNFTI-E-El M1j 890 lETKADDVDSOGN-RvT 118 907l LPIDLEEQTMGKN [9-91 KPDSPDARH-YKSAS 18a 93]jPDSPDLARHYKSASP [a [96-211 IIQELPLDNTFVC Ra F99-1 SDOGYPVTT-EVPVS Ha rl-i-0iJ1 FEVPVSVHTRPVGIQ 18I 223]1 DGGFPQRSSTAILQV n171 MI] SGTYIFAVLLACVVF 116 EKI [AMQFKLWYKTGDVPL][16 KLWYKTGDVPIE 116 [fGEIFTTGARID-REK 16 1051 HFYEEVbAILP-DEI 16 13611 APLFIPATVINISIP-E- 16 [182][ QNIFGLDVIEEGD [i-J [246][ HPVFKETEIESE 516 [28-j[ KIHFSFSNLV-SNIAR [IJ DIRYIVNPVNTV [6 L 4 RFNQLLETMAY 1He F4201TMYLDYESTKEYAJI F4231YLDYESTKEYIL Eel- I 45-0j[ AMLFIKVKDENDA Ee6 [463] APVFTQSFVTVSIPE 16! gfI[ KY'LFriLAKDNG-VPP 16 [~[VRVFVSIIDQN S F161

[HNEYNFYVPENLPH

f EYNFYVPENL-PRHG-Tl 1] :575]1 YNFYVFENLPRHG1V :596] DPDYGDNAVLSIL g~ 639 YTFYVK.AEDGGRVS-R e Table XLVIII 109P1D4v.1- DRB31 0401-15-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start sition plus fourteen 1693 1 1VVFQV1AVDNDTGM[fE Elj[ EVRYSIVGGNTRDIYF f F760][ DSLFSVV1VNLFVNE fE 12:]1 W1FITAWRCRQ-AP Eel7 [isifj NSAINSKYTLPAAVD 1s] 9531 ETPLNSKHHIIQELP~] [IlI MDLLSGTYIFVL n14] 9a- 1 IFAVLLACVVFHSGA 14 M10JI FAVLLACWVFHSGAQ] 4] D]fl AVLLACWVFHSGA1E4 flRACVVFHSGAQEKNYTj 14 [i][LKDLNLSLIPNKL RfI J[FKLVYKTGDVPLIRI 1 4 TGVLIIETGE 14 [1[DVPLIRIEEDTGEIF J141 l7fiI VPLIIRIEEDTGEIfT141 F741 IJ RIEEDTGEIFTTG 1 [gJ[ GARIDREKLCA-GIPR- 14j F171FYEVEVAILPDEIFR F[4 F1 091 EVEVAILPDEIFRLV 4] E PDEIFRLVKIRFLIE I4 FI-1-91 IFRLVKIRFLIEDIN 41 1 2-6 1 RFEDINDNAPLFP 114 1~f ATAlNISIPENSAN n14 F14flj NISIPENSAINSY 141 ETi~j PMAVDPDVGINGV-QN 14 Eiflj INGVQNYELIKSQNI I F_75_ NYELIKSQNIFGHLD-V [4 [j7oJ[ YELIKSQNIFGLD-VI If [ifl ]GLDVIETPEGDKMQ ff4 18]1 LD1ETPEGDKMPQL i:1 fGDKMPQLIVQ KELDR14 QLIVQKELDRE EKIDT 141 204] QKELDREEKDYVK R4 213 D YV M K-VKV ED-GG-F-P 14 216VMKVKVEDGGFQRS R,4 00 Table XLVIII- lOOPD4v.1-1 ORBI 0401-15-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is amino acids, and the end position for each peptide is the start position plus fourteen I F25-1 ETEIEVSIPENAPVG [4 255]1 EVSIPENAPV SV R14 261 J NAPvGTSVTQLHATD 1 4] 2~8[ FSNLVSNAR FHL 41 299 LFHLNATTGLITIKE] n14 305[ UTGUTIKEPLDR 1 F37 1(KLLVLASDGGLMPAR JH4 32fl][LLVLASDGGL-MPARA JH F339JAMVLVNVDVNDNVP F34] ][NVLVNVrDVND~~ 4 34211 LVNVTDVNDNVPSI-D ft 367]1 TWLSENIPLNTKIA 1H41 37111H SENIPLNTKIALITV [4J 41511f. QFLLETMAYLDYEST 4 43111 EYAIKLLAAD)AGKPP [4( 43-31 AIKLLAADAGKPPLN1 14 F434]1 IKLLAADAGKPPLNQ I4] 44311 KPPLNQSAMLIV 41 F4-]81 QSAMILFIKVKDENDN H4 [fi[f FIKVKDENDNPF E14 F46]2 NAPVFTQSFVTvsip1 H14 F46]8 IQSFVTVSIPENNP [l4] F47]0 FV1VSIPENNS=PGIQ H4 F48]0SPGIQLTKVS-AMD-A-D [4 F50]2 NYLLGPDAPPEFSLU -D 14 51911 TGMLTVVKKLDRE1E41 525]1 VKKLDREKEKYLFT 14 53811 IFTILAKDNGVPPL-TS fj4 553]1 NV1VFVS1IDQDS1] 58611 HGTVGLITvTDY F14 588] TVGLrrvTDDYGDN- 11 5911 LrTVDPDYGDNSAV 114] 602l NSA vTLSILOENDDF4]H i~1 AvTLSILDEND-DFTI [14 K7]1 LSIWDENDDFIQ 11M Table XLVIII 109P1D4v.1- DRBI 0401-15-mers Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 15 amino acids, and the end position for each peptide is the Istart position usfrte [6221 TGVRPNISFDRE-KQ W RPNISFDREKQES-YT [4 [6-56[SAKVrlNWDV NNK E[4 76-11 TINWVDVNDNKPVFI E4 F6-63[ WDVNDNKPVFIV-PP 7 j[ NKPVFIVPPSNS YE 4 6711i PVFIVPPSNCSYELV 41 6i8]1 SYELVLPSTNPG1W [4] 6i3] ELVLPSTNPGTVF 4 70811I NAEVRYSIVGGNR 4 713]1 YSIVGGNTIRDL FAID[4 7301 TGNITLmEKCDVD [4 z73 j ILMEKCDVTLL [4 7411 VTDLGLHRVLVKAD [H 7731 NESvi-NATILINELVR [4 783] NELVRKSEAVP 141 824] VVVFITAVVRC-R-Q 41 831 ITAWVRCRQAPK 141 861]1 NSRQMIMMKKKKKI( 1 ]I FvTiEETKADVS 141 913] EQTMGKYNwvrTPTT 14 91 9I YNWVTTPTTFKPS 14 932j SPOLARHYKASQ F141 970] NTFVACOSISKCS R14 988 PYSVSDCGYPVF [41 10001 TFEVPVSVHTRPG H4 1002 EVPVSVHTRPVGIQV [RU Table XLIX- 109PID4v.1-DRBI 1101-15-mers J Each peplide Is a portion of SEQ ID NO: 3; each start position Is specified, the length of peptide is 15 amino aclds, and the end position for each peptide Is the start position plus fourteen :5:51 KYLFTILAKDNGVPP 2 827 1VIFAVVRcRQAI- [Table XLIX- 1091D4v.-ORB1 1 101-15-mers Each peptide is a portion of SEQ1 ID NO: 3; each start position Is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the Istart psltion lsfute [16PDEIFRLVKIRFLIE [25HFSFSNLVSNIAR-RL]F2 [i0-001 TFEVPVSVTPG ]JAMQFKLWKV PL[4 518 IRTGmLr1VKKLDREK E3~ 5191 TGML1WQKLDREKE [23 882] LLNFV11EETAD [23 289] SNLVSNIARRLFL 22 :MD QESYTFYVKAEDGGR E22 73011 TGNrTLMEKCDVTDL]22 779]1 ATLINEILVRKSE 22 10021 EVPVSVHTRPVGIQ-V [2 :1 2:]lVLLACWVFHSGAOQEK Fji :3:7]l NVLIGDLLKDLNLSLJ F2' E2Z I LVNVTDVNDNVPSID [21] 5221 LTVWKKLDREKEK 21] 808 SDYKILVAGTI [21] 861]NRQMIMMKKKKKK [1 1:1 AVLLAC VVFH SGAQE g0 82] GEIF1GARIIREKL 105 IHCFYEVEVALPDEI] 2O] 12]KDTYVMKVVDG 9 :2][fl GTSVTOLHATDADIG jIJ :29I] SNLARRLFHLNATTG I~ 7D II NSPGIQLTVA A 9I :482 GIQLTKSMDADSG I2i 647 ]AEDGGRVSRSSK N~ E32][SPDLARHYKSASPQ-PINI :972][FVACDSISKCiSSSSS [N E31 APLFPATVINISIPE [19] ~14 IFGLDVIETPEGDKM [19 29 [ARRLFHLNATTGUIT 5-9 121NHKLLVLASDGGL-MP APVFTQSFvTvsiPE J 9] 6:0D TINWVDVNDNKPVF-l jF19 7201 TRDLFAJDQETGNIT jfH 8211 TITVM/V]FrTAvvR R~ 00 00 Table XLIX- 109P1D4v.1-DRB1 I 1 101-5-mets Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide is amino acids, and the end position for each peptide Is the start position pus fourteen 7-I 1YIFAVLLACWFHS 18 71f! DVPLIRIEEDTGE1 18 126 RFUIEDINDNAPLFP 18 555- NSKYrLPAAVDPDV 18 182 QNIFGLDV1ETPEGD 18 213-f TVMKVKVEDGGFP 18~ 3791 KIALITVTDKDADHN 18 431f EAKLAAGKPP 18 rifI LTKVSAMDADSGPNA [49'8 NAKINYLLGPDAPE1 Fj1-01 PPEFS.CRTGMLT 18 I 5861 I-GTVGUTVTPDYG g~ F6951 VFQVIAVDNDTGMNAI f8 7661 DSLFS1VNLFVNE 18 Nil SWIVNLFVNESVTN 18 [7971 NTEIADVSSPTSDYV

H~

993] DCGYPVrTFEVPVSV 18~ 104-4 EHCFYEVEVAILPDE 17J 117l DEIFRLWIRFLIEI) 17 210 EEKDTYVMKVI(VEIJG 17 246 HPVFKETEIEVSIPE 17 380 (AlM DAHG1 449i SLiKDENDNA 17 638 TYKEDGGRVS [670 KPVFIPSNCSYEL7 [6931 TVFQVIAVDNDTGM 1 7-4-4 LGLHRVLVKANDLGQ

I

819 AGTITVVIFV 17 986 SPYSVSDCGYPVT 17 138 LFPATVINISIPENS 16 r17-3 VQNYELIKSQNIFG L 16 399 FTDHEIPLP n16 450 AMLFIKvKDENDNAP 16 467 TQSFVTVSIPENNSP 1 n16 554 TVSIDQNDNSP

MIA

Table XLIX -109P1 04 v.1-RBi Each peptide is a portion of SEQ ID NO: 3; each start position is specified, the length of peptide Is 15 amino acids, and the end position for each peptide Is the start posillon plus fourteen 61 IDSQTGVIRPNISFD 16 679 NCSYELVLPSTNPGT 16 68] TNPGTWFQVAVDN 1i~ 1704 IDTGMNAEVRYSIVGG 16 710I EvRYSIVGGNTRDLF 16t 738 KCDVTDLGLHRVLVK 16 761VNLP/NEsvTNATUI 16 F80 ]TSD-YVKJLVAA-VAG-T 16 [91-6 MGKYNTTTF 16 936ARYKSASQPAFQI 6 Table M1XI-1 09P1 D4 v.2 C' Terminal-Al 9-mers Each peptide Is a portion of SEQ ID NO:- 5; each start position is specified, the Plength of peptide Is 9 amino acids, and the end position for each peptide Is the start position plus eight 8 PISTSI16 12 RSTIEICS 10 10 DSRTSTEI 8E 14ilCE 8 ~I 109PID4v.2 Terminal-A0201 Each peptide is a portion of SEQ ID NO: 5; each start position Is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position lus eight 10 SRTSIEI] [jHTRPTqSRTF 61j Table 109P1D4 v.2 C' Terminal A0203-9 mers No Results Found.

FTabIeXXV-109P104 v.2 C' Termlnal-A3 9-mers; Each peptide is a portion of SEQ ID NO: 5; each start position Is specified, the length of peptide Is 9 amino adcds, and th ed position for each peptide is the start positio lS eight 3 SfDS W VV1 P 4J VHIRPJISR 9jj [~~S1E9 E 8 TabfeXX(VI-109PlD4~ v.2 C' Termlnal-A26 9-mers 00 00 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of pepide Is 9 amino adds, and the end position for each peplide is the start position plus eight [8 ITFsRTKw- STI i] HPTDSRT -T1 202 Table )OWIII 109P1D4v.2 C'erminal-B308 9-mers Each pepide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptde Is the start position plus eigt 1 ST C E14 Table XXIX 109P1 D4v.2 C' Terminal-Bi510- 9-mers Each peptide is a portion of SEQ tl) NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end positon for each peptide Is the start position pus eight gJ TRPTDSRTS F4j Each peplide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptie is 9 amino acids, and te end position for each peptide is the start position plus eight 11 SRTSTIEIC 13 W VHTPTDSR 12 EF1TR-PTDSRT-S E1 Table MXX IO9PID4v.2 C' Termlnal-132709 9-mers Each peptide Is a portion of SEQ NO: 5; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position I lus eight Each peptide is a portion of SEQ NO-. 5; each start position Is specified, the length of peptide Is 9 amino adds, and the end postion for each peptide is the start position pus eightI 1 SRTTIEIC 12 D6jjTRPTDSRTS 1 14 STECI1 10 DRTT EjJ Table Iii M0P1 0.2 Crrerminal-B308 9-mers Each peptide is a portion of SEQ ID NO- 5; each start position is specified, the length of peplide Is 9 amino acids, and the end position for each peptlde is the start position Plus eight 00 00 0 Each peptide is a portion of SEQ ID NO 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight SSTIEICSEI 13 PTDSRTSTI 12 DSRTSTIEI 11 Table XXXV 109P1D4v.2 C'Terminal-A0201-10mers Each peptide Is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 8 RPTDSTSTI 0 l TDSRTSTIEI 10 Fi1 RTSTISCSE 10i 01 TSTIEICSEI I 4 SVHTRETDSR 6 HTRPOSRTS 8 7 TRPTDERTST 6 TI VPVSVHTRPTI 5 Table

XXXVI

109P1D4v.2 CTerminal- A0203-10mers No Results Found.

Table XXXVI 109P1D4v.2 C' Terminal-A3-10-mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 4J SVHITRPDSR 17 2J PVAVHIEPTD ~RP [DSRTSTI 12 6 HTEPTDSRTS Table XXXVIII 109P1 D4v.2 C' terminal-A26-10-mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 13 RTSTIEICSE 13 4ESVHTRPTDSR12 11 DSRTSTIEIC 12 W PVSVHTRPTD 1 1 HTRPTDSRTS 1 F PTDSRTSTIE Table XXXIX 109PID4v.2 CTerminal-B0702 Each peptide is a portion of SEQ ID NO: 5; each start position Is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nie 1 VPVSVHTRPT 18 RPTDSRTSTI I 10 TDSRTSTIEI 9 Table XL- 109P1D4 v.2 C'Terminal 608-10mers No Results Found.

Table XXXIV 109PiD4v.2 C' TerminaAl-l10mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amIno acids, and the end position for each peptide Is the start position plus nine f PIDSRTSTIE 16 EHIRPTDaRTS 00 00 Table

XI--

109P1 04 v.2 C Terminal- 81510- No Results Found.

Table

XLII-

109P1 04 v.2 C' Terminal 82705- No Results Found.

Table 109P104 v.2 C' Terminal- 82709- I 0-mers No Results Found.

Table XLIV I 09P1 04v.2 Cterminal-8442- Each peptide is a portion of SEQ ID NO: 5; each start: position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine TSRS1E12 Table

XLV-

109PID4 Q. C' Terminal- B35101- 10-iners INo Results Found.

Table XWVI-1OOP1B4v.2 C' Terminal-DRBI 0101 15-niers Each peptide Is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide Is 15 amino acids, and the end position for each peptide is the start position plus fourteen Mj]TFEVPVSVHTR PTDS H []SVHTRPTDSRTSTIE Ig 16]VPVSVHTRPT DSRTS gP [Ij IITRPTDSRTSTIEIC [ig [4]FEVPVSVHTRPTDSR11] W lPVSVH-TRPTDSRTSTR HI1 RPTDSRTSTIEICSEIMP [fliE'/PVSVHTRPTDSRT Table XLVI I-I 09P1 D4v.2 C~ Terminal-ORBI1 0301 Each peptide is a portion of SEQ ID NO: 5: each start position Is specified, the lenth of peptide Is 15 amino acids, and the end position for each peptide Is the star I position plus fourteen MI~ VH-TRPTDSRTTE 1177 I 1EVPVSVHTRTD T gP jfo ]VTTFVPVSVHTRP1T1E Table XLVIII-1 09P1 D4v.2 C' Terminal-DRBi 0401 Each peptide is a portion of SEQ ID NO, 5; each start position is specified, the length of peptide isiS5 amino acids, and the end position for each peptie is the start position plus fourteen E] V1TFEVPVSi-TPT [22 E4]FEVPVSVHTRPTDSR [18 101 VHTRPTISRTSTII 1-8 [fl TFEVPVSVHTRPTDS [4 [f]jEVPVSVHTDSRTE[4 1M] SVHTRPTDSRT-ST1--E [2 H1 HTRPTDSRTSTIEIC [2 RPTDSRTSTIEICSE [12 Table XLIX-109P1 D4v.2 C' Terminal-DRB1 1101 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the lengtho peptide is 15 amino acids, and the end position for each peplide Is the start position plus fourteen qs]EVPVSVHTRPTSRjle VTTFEVPVSV-TRPT ff v.2-N' termInal-Al -9mars Each peptide Is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide Is 9 amino adids, and the end position for each peptide Is the start position plus eight 1~]QIFQVLICGL 1 00 00 15 VLCGLIQQT 22 L7 WVMQIFQV 20 18 GLIQQIVTS 19 f24 VTSVPEMDL W1 16 LCGLigQTV 1i4I 22 QTVTSVPGM W1 TSVPGMDLL 1 2 RTERQWVLI 13 9 uQlFQVLC [3 Table XXIII-109P1D4 v.2 N' terminal- A0201 9-mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino adcids, and the end position for each peptide is the start position plus eight E 14 i1 LIQQTfSV iii QIFQVCGL 2 8 VOLIQQ. 2 1 VLCGLQQT 7 WVLQIFQV 18 GLIQQIVTS 1 24 VTSVPGMDL 16 1[ LCGLQQTV 14 []QTVTSELPGM [41 TSVPGMDLL 14 [1RTERQW VLI 9 LIQIFqVLC 1 Table 10910 v.2 N' terminal- A0203 9-mers No Results Found.

Table XXV-109P1D4 v.2 N' terminal-A3- 9-mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight !1 QVLCGU.QQ 19 8] VL!QlEgVL 17l terifialA2-9-er 7 WVLIQEQV 16 26 SVPGMDLLS 1 VLGL!QQT 15 23 TVISVeMD 14 W LglFS .C 13 29 GMLLSGTY 12 2 RTERQWVLI 11 11 QIFQVLCGL 11 19 LIgQTVTSV 1 Table XXVI 109P1D4v.2 N' terminal-A26-9-mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight 11 QIFQVLCGL 20 24j VTSVPGMDL 17 T] ERQWVLUQI j 14 QVLCGLIQQ 22 QTVTSVPGM 16 7 WLIlFQV 15] 23j 1VTSVPGMD 81 VUlQIFQVL.1 2j TSVPGMDLL 14 RQWVLIQIF3 Table XXVI 109PID4v.2 N' terminal-A26-9-mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 2 GMDLSGTY 1 26 SVPGMDLLS 12 MRTERQWVL 11 Table XXVII- 109P1D4 v.2 N' terminal-B0702 9-mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 24 VTSVPGMDL 16 27 VPGMDLLSG 1 8 VLIQIFQVL 12 1 MRTERQWV 11 25 TSVPGMDLL 11 11 QIFQVLCGL 2 RTERQWVLI 9 VLCGLIQQT 8 1 CGLIQQTVT 8 1 LIQQTVTSV 8 [2 QTVTSVPGM 8 Table XXVIII 109P1D4v.2 N' terminal-B08-9-mers 00 00 0 Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight [lMRTERQWV 20 VQIFQVL 17 1 QIFQVLCGL VTSVPGMDL 12I

TSVPGMDLL

Table XXIX-109P1D4 v.2 N' terminal-BWiO 9-mers Each peptide Is a portion of SEQ ID NO: 5; each start position Is specified, the length of peptide is 9 amino adcids, and the end position for each peptide is the start position plus eight TSVPGMDLL I1 I MRTERQWVL 1 j VLQIFQVL 1 4 VTSVPGMDL 1 QIFQVCGL I I RQWVLIQIF 22 QVTSVPGMj 8 Table XXX-109P1D4 v.2 N' terminai-B2705 9-mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight Table XXX-109P1D4 v.2 N'terminal-B2705 9-mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino acds, and the end position for each peptide is the start position plus eight 1 MRTERQWVL 41 EROWVLlQ i20 RQWVUQF 1 QIFQVLCGL 17

H

8J VLI FQVL 16 ~]GMDLLSGTY 1L5J aj5 TSVPGMDLL 1f4J Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 9 amino adds, and the end position fbr each peptide is the start position plus eight 8 VLIQIFQVL 16 STSVPGMDLL 4 ERQWVLIQI 5 RQWVIQIF 1 QIFQVLCGL 2GMDLLSGTY 13 Ea MRTERQWff2 3 TERQWVLIQ I RTERQWVLI I VTSVPGMDL 11 p o IFQVLCGLI 9 Table XXXIId 109P104v.2 N' termina-85101- 9mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position plus eight 4 ERQWVLIQI 14 19 LIQQTSV 27 VPGMDLLSG13 1 MRTERQWVL fI IFQVLCGL 12 16 LCGLIQQTV

CGLIQQTVT

2 RTERQWVU 11 SWVLIQFQV II 8 VUQIFQVL 11 11I QIFQVLCGL l12 2 IQQTVTSVP 8 2PGMDLLSGT 8 Table XXXII 109PiD4v.2 N terminal-84402-9 mers 00 00 Table XXXIII 109P1D4v.2 N' terminal-B5101- 9mers Each peptide is a portion of SEQ ID NO: 5; each start position Is specified, the length of peptide is 9 amino adcids, and the end position for each peptide is the start position plus eight 24 VTSVPGMDL 7 TSVPGMDLL L Table XXXIV 109P1 D4v.2- N' Each peptide is a portion of SEQ ID NO: each start position is specified, the length of peptide is 10 amino adcids, and the end position for each peptide is the start position plus nine

I

2 RIERQWLIQ 3 TIVPGMQLLS 16 28 PGMDLLSGTY 15 29 GMDLLSETYI 11 N Eoss llB Table XXXV-109P1D4 v.2-N' terminal-A0201- 1 0-mers Each peptide is a portion of SEQ ID NO: each start position Is specified, the length of peptide is 10 amino acids, and the end position for each peptide Is the start position plus nine 18 GLIQQIVTSV 29 VLCGLIQQTV 5 1j IQIFQMLCGL l8 iE 1 I Q1FQVLCGLI L7 29 GMDLLGTYI [7 Table

XXXVI-

109P1D4 v.2-N' terminal- A0203- 10-mers No Results Found.

Table XXXVII 109P1D4v.2 N' termnal-A3-10-mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino adcids, and the end position for each peptide is the start position plus nine 7]] 26 SVEGMQLLSG B 7 WVIQIEQVL lI VL!QIEQVLC 1 14 QVICGI.aQQT I17 15 VLGL 1QQTV 6 18 GLIQQ]YTSV 16 19 LIQTYSVP 23 TVSVEMDL 14 Table XXXVII 109P1D4v.2 N' terminal-A3-1 0-mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 9 LIqIFQLCG 12 28 PGMDLLSGTY 12 11 QFQVLCGU II1 17 CGinIQQ TS 11 2 RTERQWgLIQ Table XXXVIII 109P1D4v.2 N' terminal-A26-10-mers Each peptide is a portion of SEQ ID NO: 5; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine ERQWVLIQIF 23 TVTSVPGMDL 2 7 WVLIQIFQVL 18 26 SVPGMDLLSG 17 10 IQIFQVLCGL 16 2 VTSVPGMDL 16 14 QVLCGLIQQT 1 fE2 Q1VTSVPGMD 1J 2 1RTERQWVMIQ 3 28 PGMDLLSGTY J1 Table X)OXXIX 109PiD4v.2 N' terminal-BO702-1 Omer 00 00 portion of SEQ ID) NO: F ahppiei5; each start position Is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position Pus nine 27 VPGMDLLSGT 17 7 WVUQIFQVL 12 [g IQIFQVLCGLIF11 N' TVT-SVPGMDL 10 16 LCGUQQ1Vr 9 F]j MVRTERQWVU1 8 ]TERQVLNQl 8J VLCGLIQQ1V J 18 GLIQQTVTSV ITable

XL

N' terminal-, B08-l0mers Table )XLI 109P1D4v.2 N' terminal- I Omer No Results Found.

Table XLII N' terminal- B2705- No Results Found Table XLIII 109P1 D4v.2 N' terminal- B2709l0mer No Results Found.

Table XLIV 1O9PID4.2 N' teina184402-l Omer Each peptide is a portion of SEQ ID NO: 5; each start position Is specified, the length of peptide Is 10 amino acids, and the end position for each peptide is the start _position plus nine 31 TERQ\WLjQI1 21 41 ERQWVLIQIF 7 nVIIQ 13 11 QFQLCL 111 Table XL-V 1O9P1D4v.2 N' terminal- 85101l0mer No Results Found.

Table XLVI-IO9P1D4v.2 N' terminal -DRBI 0101 Each peptide is a portion Of SEQ ID NO: 5; each start position Is specfied, the length of peplide is 15 amino acids, and the end position for each peptide Is the start position plus fourteen :27] VPGMDLLSGTYYIFAV 3 2:QT~SPMLLG3 N' terrninal-ORBI 0101 Each peptide is a portion of ISEQ ID NO: 5; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen 4ERQWVUQIFQVLCG [iIQIFQVLCGLQV F6j f 1 RQVLIQIFQVL CGL g~S El~ FQVLCGLIQQ1TVrSV 24 HiII VLCGLIQQTVTSVPGI 3 MIJ LCGLIQQTVTSVp-GM] E23 s]j LIQIFQVLCGUQQT lg E i I C lQ TVG-M-D D8 VLIQIFQVLCGLIQQ 1I] f-TG -X-VIII-109P11)Z! 2 N'l Oterminal-DRBI 0401-% 00 00 Each peptide is a portion of SEQ ID NO: 5; each start sition is specified, the length of peptide Is 15 amino acds, and the end position for each peptide is the start position plus fourteen

FQVLCGIQQVTSV

I ERQWVUQIFQVLCG F] F1i1] IQIFQVLCGLIQQTV,~ RI] QWVLIQIFQ V-LCGLI fI LIQIFQVLCGLI-QQT La [27J QQ1TSVPGMDLLSGLa J VPGMDLLSG1IFAV

I

W TERQWVLIQVLC F18 i QVLCGUQTSVP F18 [DI RQWVLIQIFLCGL 141 [Il WVLQIFQVLCGLIQ 14! Ai IFQVLCGu Qrs 141 1J LCGLIQQTVSVP-G-M14 I1 CGLIQQTVTSVPGMD 14 GMDLLSGTYIFAVL F Table XXII-109P1D4 v.3-Al-9-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight [fl TSHGLPkGY E2 S EQASAc 23 5NTQEC 21 62 ISSDGGLGDH 19 69 DDAGSTS 1 11PIRIEGDGNSD 1 10 NSDPESIFI Is1 SIhPGKK [18 PLGYPQEEY[8 i ESTFIPG 1f 37 KSEGKVAGK 116 61 ESSSDGG8GD S 12ASDNCTEC Es 5 1~JQESATS9FY 1 302 YTMSERJHP 15 1311 P~DDSIVZ 1 [~fPqEEYFeRA I4] [14HSDACW~P I~4 [lj MsERLHSD I~ sLDHss~sQs [I3 186VIQTIALCH 13 181 VTTIALCH 13 256 seLPOVIL 13 Table XXII-19P1D4 v.3-A0201-9-mers Each peptide is a portion of SEQ ID NO: 7; each start positon is specified, the length of peptide 15 9 amIno acids, and the end position for each peptide is the start pstopuseight Table XXIII-109P1D41 v.3-A0201-mers Each peptide is a portion of SEQ ID NO: 7; each start position Is specified, the length of peptide is 9 amino acds, and he end position for each peptide is the start position plusegh 74 SLTSTHGL 0 1 ALHHSPPLV II1GLCSVDQGV jJRLHPSDDSI 203ALCHSEPPI 256SPLP 21QGADGLCSV 238SALCYPPL 166SALCHPPL 10IALCHPPV (j9 214SAHPPL 1 2A1 HSEPSA E1 15I HTRPPMIKEV 8 A MISHSSPL jj8 1IISHSSPPQV I18 LaISQAQSSVSL 18 121 AEETV7PTV 110CLIYGISDA ((7 117DACWMPASL 781 STQHHSPRV17 1iIALHSEPVT I? 53 HLPEGHQES 11 FIPGLKM 124 TVPT)EEA Ij~ 29ALCYSPPLA 16 272 VSLQGWV16 SSIKVIPTT 16 316 KVIPITFT 1 42 F6 GLGDHDAGS J 112 TFIPGLKKAIrs 21VIALHESQA IS EfJ TSERHPS 141 V6 1 SQRRVIPHL 1~ 00 00 No Results Found.

Table XXV-109P1D4 v.3-A3-9-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start ition us eight L] SVTREPe x 22 ~]LVAT~J-H [2 263ALRSQS 41 1)KVAGK~RR 31 KVPLTTFT F1 KS1GKVGK H R8 P-QV AE1SQ 301RWHPSSI 19 Ill STEIPGLKK 1 1 13P[SQASIQH 1 911 ALRHSVT 1 27 7LE L1 R314 l V Ej 185 EVQIL 7 221 PLQAH 17 245 PLAQAAIS 261 vLHB1A 7i 33 QPqRKGK 81GLLQE1 83 )PLGYPEY 16 154 SEHSS 16 [2121QVASP L 1ALIHSeeSA 11411 LIYGHSDAC 1 Table XXV-109P1D4 v.3-A3-9-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight ~JSAqASACY 12 EERSCJPM 40 1GKIAGIQR I 49 RFHLEEG B 52 FH1PEGSQE H r6i6GLDHQAGS 122EIIVIVE 62QAASCH 17ALCHSELS I4 203ALHSEPI 21 5 IASEL 239ALSEELA 45 F1 KSqRRMFH 53 HLEG~ES 92 1FDAT 3NR 12 VPTEA 1 1 TLCHSPP 117PVIQTILC13 Ii]TILCPP SRSAQVS 1 28SVDQGMqGS 13I 30 LPSDDSIK 1 Table XXVI-10P1D4 v.3-A26-9-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 9 amino adids, and the end position for each peptide is the start pitoplseIght Table

XXIV-

109P1D4 v.3mers 00 00 121 ERSCTPM 31 DDSIKVPL t~IDACWMPASL L F7 [11 STFIPGLKK 16 24TVQP 6EA 256SPLPQ VIA 16 260QVIALHRSQ[1 133DSIKVIPLT iI 136KV1PLTTFTIiI 3351DSPMEEHPL 49 1RVTFHLPEG5

EYFMDRATPS

SEITVQPTVE I~ 136 CTQEClYG RVTQTALC15 288 S VQGS f231 STMEIWIH 4 j24J 1TMEIWIHP 4 F271 EIW1EPQPQ ii ESTFIPGLK 1 11 PRVTQTIAL 18QTIALCHSP 14 Ii1PPVTQTIAL F4 J2jQTLALCHSP 4 PPPIQVSAL[4 1I1AAISHSSPL 14 (3TTFTPRQQA14 [s0 VTLPEGS 1 [6J ESSSDGGLG 1 [88 TSTSHGLPL13 [fl STSGLG 3 TSHGLPLGY 13 STVEEASDNC13 1311 EASDNCTQE 13 2 DGLCSVDQG3 3 SVHTRPPMK 131 WRSCTPMK 221 ESTMEIWI2 39I EGKVAGKSQ 56EGSQESSSD 2 71DAGSLTSTS2 Table XXVI-109P1D4 v.3-A26-9-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 109 PESTFIPGL t2J 123 ITvQ VE 1 130EEASDNCTQ 135 NCTECUY 1 139ECLIGHSD 12 22QVSALHHSP [2 234SAQASALCY Table XXVII- 09P1D4 v.3-80702-9-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 22PPSAQASAL 2 256 SPLPQVIAL 23 [i61j PPVTQTIAL [~JPPPIQVSAL 2PPLVQATAL 18 TPKESTVM1 7SPRVTQTIA 19 2OJSPPPIQVSA11 24 98 HPSDDSIKV 18 171 SPPLSQAST17 19]5 SPPVQTIA 7 21'1j SPPLVQATA J7 2311 SPPSAQASA 7 Table XXVII-109PD4 v.3-10702-9-ers Each peptide is a portion of SEQ ID NO: 7; each start position Is specified, the length of peptide Is 9 amIno acds, and the end position for each peptide is the start position plus eight SPPMKEWRS F f7 RPPMWR [4j (76 TSTSHGLPL j4 1141E IPGLKKME 14 i5 CHSPPVTQT 127HHSPPLVQA[4 32DDSIKVIPL ~38IPLTTTPR 14 gi SQRRvrFHL g 96 10 PESTFIPGL j~fHHSPPSAQA j~ 1211CYSPPLAQA E1 21 ISHSSPL 1SQAQSSVSL SHTRPPAKEV 12 35-1 12PQK E 3 54 ILPEGSESS 159 1 82J LPLGYPQEE J3 81DPESTFIPG 160SSQAQASAL [jJSALCHSPPL 1 9 CHSPPLSQA12 1 PRVTQ A 251CHSPPPIQV [2j 214 SALHHSPPL [U 231SALCYSPPL L2 [2SHSSPLPQV[2 251LPQVIALHR 12 v.8-98ers 00 00 Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 312 DDSIKVIPL 2 256 SPLPQVIAL 2 11 IPGLKKAAE 46 SQRRVTFHL 18 74 SLTSTSHGL 18

PPPIQVSAL

220PPLVQATAL 1i 7 RPPMKEVR17 115 PGLKKAAEI 17 16 GUKAAEIT 1i 1 PPVTQTIAL [7 232 PPSAQASAL 4 SIKVIPLTT 17 33 QPQRKSEGK 6 44 GKSQRRVTF 16 6 SALCHSPPL 16 1 SALHHSPPL16 8 SALCYSPPL 1 SSPRVTQTIA 15 39 IEGKVAGKSQ14 961 TPSNRTEGD14 147 DACWMPASL 1 AAISHSSPL 14 ALHRSQAQ 14 9 PMKEVVRSC 1

SSQAQASAL

PPLAQAAAI 1 SQAQSSVSL 1 19 PMKESTTME 1 SDNCTQECL 12 231ALCHSPPPI 1 RLHPSDDSI 12 3241TPRQQARPS SQRKSEGKVA 1 SKSEGKVAGK 1i PESTFIPGL 11 84 PRVTQTIAL 11 Table XXVIII-109P1D4 v.3-B08-9-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start osition plus eiht [278 GWVQGADGL 1 Table XXIX-109P1D4 v.3-81510-9-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start posItion plus eight 30 IHPQPQRKS 16 217 HHSPPLVQA 16 180 QHHSPRVTQ 193 CHSPPVTQT 205 CHSPPPIQV 169 CHSPPLSQA 14 181 HHSPRVTQT 216 LHHSPPLVQ 229 HHSPPSAQA 14 25 SPLPQVIAL1 7 SQAQSSVSL ff1 [4 GKSQRRVTF f1 09 PESTFIPGL GHSDACWMP 13 LHSPPSAQ I13 253 SHSSPLPQV 27GVQGADGL 13 4 VHTRPPMKE 52 FHLPEGSQE 12 69 DHDAGSLTS 12 156 DHSSSSQAQ 1 208 PPPIQVSAL 2 22 PPLVQATAL 12 2 PPSAQASAL 2 Table XXIX-109P1D4 v.3-B1510-9-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 300 QFYTMSERL 312 DDSIKVIPL J1 59 QESSSDGGL 11 76 TSTSHGLPL 79SHGLPLGYP GNSDPESTF 1 147 DACWMPASL 1 16 SSQAQASAL i 66 SALCHSPPL 84 PRVTQTIALE 1 PPVTQTIAL 1 1 SALHHSPPL 1 SALCYSPPL 46 SQRRVTFHL 67 LGDHDAGSL IJ 74 SLTSTSHGL SDNCTQECL

AAISHSSPL

6 LHRSQAQSS 1 LHUPSDDSIK

IKVIPLTF

DSPMEEHPL

[IITPMKESTTM 9 2 VSVHTRPPM 8 84 LGYPQEEYF 8 RPSRGDSPM_ 8 Table XXX-109P1D4 v.3-B2705-9-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 00 00 2 184 PRVTQTIAL jj0 (GKVAGKSQR 19 278GVGDL1 281 IW1HPQPQR 18 41 KVAGKSQRR 118 7 RPPMKEVR17 37 KSEGKVAGK 44 GKSQRRVTF 17 111 STFIPGLK 17 315IKVPLTTF 7 48 RRVHLPE 16 99 ENRTEGDGNS6 GNSDPESTF 1 HRSQAQSSV6 26SQAQSSVSL 330RPSRGDSPM 1 18 IITPMKES1TM 1 193DRATPSNRT EI1 129PPIQVSALH 15 220PPLVQATAL 2501AAISHSSPL 5 [2JSPLPQ VIAL 1 2SQFMMSER 1 0Q1MSERL5 38IPL1TFTPR s 72 1AGSLTSTSH14

PESTFIPGL

11 PGLKKAAEI 1 [6jSALCHSPPL 1 ~7JPLSQASTQH 4 W71ASTQHHSPR 4 24SALHHSPPL 14 4SALCYSPPL ~jERLHPSDDS 4 307RLHPSDDSI 3] RGDSPMEEH14 TRPPMKEW 13 141 VRSCTPMKE 1 2j11 STfMEIWlH 1 29 WIHPQPQRK113 KsQRRVFJ 62SSDGGLGDH Table XX-109P1D4 v.3-827059-mers Each peptide is a portion of SEQ 1D NO: 7; each start position Is specified, the length of peptide is 9 amIno acids, and the end position for each peptide is the start position pluseight M][LGYPQEEYF E3 92FDRATPSNR 13 137 TQECYGH 258LPQVALHR 3 312DDSIKVIPL 13 TFTPRQQAR 2] 3L 3JSRGDSPMEE j3 12 EEVVRSC TPM 33 IQPQRKSEGK I IQRKSEGKVA 11? 59 IESSSDGL 67 LGDHDAGSL 78 TSHGLPLG(1 83PLGYPQEEY 86 IYPQEEYFDR 2 13SDNCTQECL 1 135NCTECUY 17DACWMPASL [2 160SSAQASAL 12 16PPVIQTAL [2 SPPPIQVSAL 12 [2jPLVQATALH 1 23 PPSAQASAL 23VQGSATSQF 1 LHPSDDSIK IM 39ARPSRGDSP [2 Table XXX-10P1D4 v.3-B2709-9-mers Each peptide Is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide Is 9 amino acids, and the end position foreach peptide is the start position plus eight [l[I 14PRVTQTIAL I~ 6 TRPPMKEW 1 265HRsas 18 48 RRVTFHLPE 16 278 SPLEVIAL 14 76 TSTSHGLPL 13 166 SALCHSPPL 13 SALHHSPPL 13 22 PPLVQATAL 3 28SALCYSPP~L3] 250 ISHSSPL 30QFYIMSERL 307RLHPSDDSI 44 GKSQRRVf FI 67 LGDHDAGSL [7J SLTSTSHGL J 99 NRTEGDGNS 190LALCHSPPV 285GLCSVDQGV ERLHPSDDS f~ 329ARPSRGDSP 330 RPSRGDSPM F051RKSEGKVA ~3 QESSSDGGL Ji [~I[LGYPEYF ~I [93I RATPSNRTI 1 lojGNSDPESTF 11 0PESTFIPGL 11 15 PGKKAAEI 11 11AETVQPV 11 SSSQAQASAL E' 19 PPVTQT(AL Ei 2 PPPIQVSAL 2PPSAQASAL [i 25:] Ell~ 1 2JSHSSPLPQV 11 267 SASSVSL1 96SATSQM 31 DDSIKVIPL 25PRQQARPSR i 00 00 Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eiht M PPLAQAI [24 R] VAG KSQRRV [23 IALCHSPPV 2 gHPSDDSIKV

E

us1 PGLKKAAEI21 256 SPLPQVAL21 208PPPIQVSAL F31SALCYSPPL Ii~ 1 SALCHSPPL 51 PPVQTIAL 1 [i]SALHHSPPL 1 ~3JPPSAQASAL 1 i IPTTFTPR 82 PLGYPQEE 17 SDPESTFIPG 1 J30PSDDSIKV1 I 7 W RPPMKEVR 1 71 DAGSLTSTS 16 I2IAAISHSSPL M E15 F211 QGADGLCSV Ej5 R9PPMKEWRS 5 118 TPMKESTM ft5 1671 LGDHDAGSL 4 RATPSNRTE S15 1 DNCTQECLI 1 1iIl PPLSQASTQ [15] L1 HSPRVTQTI 1I [i1 HSPPVTQTI 15 29SPPLVQATA 1 [4jLAQMAAISH [1s SLPQVIALHR 1 6I TRPPMKEW 1 [5j LPEGSQESS 1 [8s IYPQEEYFDR14 Table XXXIII 109P1D4v.3-B5101 9-mers Each peptide is a portion of SEQ ID NO: 7; each start position Is specified, the length of peptide is 9 amino ads, and the end position for each peptide Is the start position plus eight 117 LKKAAEITV 14 162QAQASALCH 22IALCHSPPP 24SAQASALCY HSSPLPQVI H 5IA41RSQAQ 14 282 GADGLCSVD J32DDSIKVIPL i4 22ESTMEIW1 3 1IPGU(AAE 13 1 KAAEIQP 3 20 AAEITVQPT13 121AEITVQPTV1 115SPPVTQTIA 3 11TALHHSPPS IIJ F2-216 2681 gASSL 3 6 SATSQM 13 j QFYMSERL[1 20 MKESTTME i 341 I4 IJLGYPQEEYF 1161NSDPESTFI I 2 1 EASDNCTQE 112R 17 SPPLSQAST iJ SPRVTQTIA

SALCHSPPPIJI

iJSPPPIQVSA 12PPIQVSALHIii Table XXXIV 109P1 D4v.3-AI 9fl 9s Table XXXIII 109P1 D4v.3-B5101 9-mers 00 00 Each peptide is a portion of SEQ ID N0.

7; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide Is the start position plus nine LE SISHGLPLGY 1 PSAQASALCY 15i F j DNCTQE LIY 121 IJSDGGLGDHD 18 101 ERIEGGSOP18 [071 N9DPESIFIP La

KSEGKVGKS

[3121 SEDSIKV1PL j0 83JLGYPEEY FjL [29-4VEGSAT6QFY 6 1331 A~DNCTqECL 1 Table XXXV-109P1D4 v.3-A0201-10-mers Each peptide is a portion of SEQ ID NO: 7; each start position Is specified, the length of peptide is 10 amino acids, and the end position for each peptide Is the start osiion plus nine HTRPPMKEW 16 20I PMKESITMEI i 112 STFIPGKA 16 1 IVQP1VEEA 6 li SLDHSSSSQA 1 192 ALCHSEPVTQ 312 SDv PL 74 GSLTSISH E15 14 LYGHSDACW [iJASAI-CSPPL i)h liel ALHSlPLsQ 115 Fa1ASALCSPPL 1 541 HLPEGSQESS 1 9DPESTIPGL 14 14 FIPGUKAAE 14 il] IPGLKMAEI II4 1214]( VSALHISPPL Ii0I [241 ALHRSA1SS 141 1271 RSQAQSVSL 14 13091 U-PSDSIKV 14 13351 GDSPMEEHPL 14 GLPLGPQEE 13 (101 SSSA9ASAL J~i 1141 SPRVTTIAL 13 [rsjl] IALCHSPPVT 13~ 119 SPPVTTIAL 1i3 12041 ALCHSPPPIQ 13 216si AHHSEPLvQ 13 220 SPPLVqATAL 13 127J1 TALHHPPSA L1 128]( ALHHSPSAQ Ff3 L3JSPPSAQASAL f I3ISALCYPPtA 3 J41ALCYSPLAQ Table XXXV-109P1 D4 v.3-A0201-10-mers Each peptide Is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 10 amIno acids, and the end position for each peptide is the start position plus nine [24 LCYSPPLAQA 3 SPPLAkAAAI IiHl N TSERLHPSD 13 Z2]I TTMEIWIHPQ 121 [36] WIHPQQRKS 121 134] QPQRKEGKV 121 I59] SQESSSDGGL 12 133 ASDNCTQECL 12 11371 CTQECLIYGH 11411 CLIYGHSDAC 1 11781 ASTQHHSPRV j~i 1182l HHSPRVTQTI_112

CHSPPFTQTI

l~[LCJ-SPPPIQV 12

SLHHSPELVA

12SPLPQYiIAH 262 VILE QE M272 E12W 278 lQGwV D l12 285DGLCSDQGV 12 M289 12 30 MsERL12 3 YTMSELHPS 1 3 RLHPSDDSIK j [i]HPSDDSIKVI E Table XXXVI 109P1 D4v.3-A203 lO-mers Each peptide Is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine 00 00 Table XXXVII 109P1 D4v.3-A3 e-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide Is 10 amino acids, and the end position for each peptide is the start position plus nine 3JRLIPSSIK I1 13 YRSIPMK 186 RV QTILc H 2 31 DKV17 FTP 23 192 A&SET

GVGSSQFI

216ALHSEEV 10 ALHRS-QAQSS 2 198 PVQTCH Lo 222L]LQA ILHH I 0I 24 PLQAeAsH 0I Table XXXVII 109P1D4v3-A3 1-mers Each peptide Is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide Is 10 amino acids, and the end position for each peptide is the start position plus nine PLEQ4ALHR 20 Ei ALHSPPLSQ 19 SIKVILTF- 1 37 (RKEGIAGK 18 [22ALIHSSA Q 8 28WVG01LC 181 9 H7AGISQRVTF 167 IGLDHAGSL E71 142FLIYGHSDACW

I

:SLDH1SSSSQA7 ~jJQVALHSPP I17 28 EIWIH PQR 1 29 IW!IHPEQRK 1 42 IIKVGKRRVI16 [ii ESIFIK 6 7 TRPIEWR

I

14 1( WESCIEMWKE [5 soI RVIFHEGS WI1 17 GLKAAEI1 1 Table XXXVIII 1O9PD4v.3-A26 l-mers Eahpeptide is a portion of SEQ ID NO: 7; each start position Is specilled, the length of peptide Is 10 amIno adids, and the end poiinfor eachpetd is the start position plus nine Table XXXV1II 109P1D4v.3-A26 Each pepfide isa portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptlde is the start position plus nine 1131 EVVRSCTPMK uS] 109 DPESTFIPGL 21 78 1 STSHGLPLGY 2931 GVQGSATSQF 105 DGNSDPESTF F1 DNCTOECLIYIP 1761 LTSTSHGLPL 1 STFIPGLKKA 1 SIKVIPTTF 18 91 EYFDRATPSN 1 124 IHVQPTVEEA 1JSPPPIQVSAL 16 121QVIALHRSQA

I

L~IKVIPLTFTP

[2][ESnMEIWIH 1 [25j TTMIEIWIHPQ[isI 28 16EIWIHPQPQR 13EIiVOPiVEE 2 SSPLPQVAL 2SDDSIKVPL 1 IIVHLPEGSQ 14 111 ESTFIPGLK 14' 128 PV SDNCJ1 117CTQECYGH 1 23 LVQATALHHS 1 34 DSIKVIPLT 1 TTFTPRQQAR14 F7-0 EE1DGD[3J 70 1DHDAGSLTST 3 [151QPVEEAS 1(31 129 TVEEASDNCT 13 189 QTIALCHSPP 13 201 QTLALCHSPP 13 289 SVDGVQ j 00 00 Table XXXVIII 109P1D4v.3-A26 I0-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide Is the start position plus nine SQFYTMSERL jg3 3 YTMSERLHPS [9l Table XXIX 109PI D4v.3-80702 Each peptide Is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start position plus nine

I

~I SPRVTTIAL 24 [2l SPPPIQVSAL 1 [19 SPPVTQTIAL j~ 2720 SPPLVATAL 1 DPESTFIPGL 11 23 SPPSAQASAL 21 [11 IPGL(KKMEI [91 [310HPSDDSIKV ~2I SPPLAAAAI [I~ 8~YPEE YFDRA [7 O PRKSEGKV [6 ~I J LTSTSHGPL iII ASALCHSPPL [iO 8 ASALCYSPPL 15 ~J RPPMKERS 14 1 TPMKESTTME 1 233PPSAASALC 1 250 AAAISHSSPL 4 1325TPR QQARPSR RPSRGDSPME 4 217 Table XXXIX 109P1D4v.3-10702 lO-mers Each peptide is a portion of SEQ ID NO: 7; each start position Is specified, the length of peptide is 10 amino acids, and the end position for each peptide Is the start position plus nine [JGDSPMEEHPL A [i~IJPPMKEWRSC [9 gIASDNCTQECL1[ FI6 0SSSAQASAL 1 R1 VSALHHSPPL 13 3121 SDDSIKVIPL 319IPf 13 VPVSVHTRPP 12 46 KSQRRvTFHL 12 551 LPEGSQESSS 12 3 LPLGYPQEEY [91 (97 ITPSNRTEGDG [92 (147ISDACWMPASL1i 210 PPIQVSALHH 221 PPLVQATAUI 245 PPLAQAAAIS 256SSPLPQVIAL 1 SPLPQVALH 1 Table XLII 109P1D4v.3- B2705 INo ResultsI LjFound.

Table XLIII 109P1D4v.3- B2709 1 -miers [NoResults Table XU 109P1 D4v.3- B1510 No Results Found.

00 00 Table

XLV-

1091104 v.3- B51 01 1O-mers No Results Found.

Table XINI-1 09P1 D4v.3-1 DRB1 0101-15-mers] Each peptide Is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is amino acids, and the end position for each peptide is the str position plus fourteen 32]0 SIKV1PLTFRQ Q 1M0 :53] QRRVTFHLPEGSQES [2g All CLIYGIISDACWMPAS N2~ 2451ALCYSPP r216I :2:1 LQQGWVQGALS 2 5] 3:3]rEIWHPQPQRKSEGK 24] 24] :216] PIQVSALHHSPPV F24] :2:31 HHSPPLVQATALHHS1 24] RI LPQVIALHRSQQS [4 j VALHRSQAQ SSVSL[2 2 j[QGWVQGADGLCVQj4 J DDSIKV1LTWfTP R [2 E lCTPMKESTTMEIIH [23] MF TQTIALCHSPPVQ [23] j TQTALCHSPPPIQV] 237 J QSSVSLQQIGWQA [25 Lgz[TTTPRQQPSRGDE23 w FEVPVSVHTRPPMKE E2 f] PQPQRKSEGKVAGKS] 2 Ii~1 PASLDHSSSSQAQAS 2 [-YSPPLAQAAJSHSs 122 [261 SSPLPQVIALHRSQA 22 g] DQGVQGSAT-SQF& M E2 1261 AAEiTvQPTVEEASD][21 :MJ] SVDQGVQGSATSQFY]21 D305 [SQFYTMSERPD [2-1 H[PPMKWREMKE EC T ablIe XLVI -1 09 P D4 v. 3 -1 DRBi 0101-15-niers Each peptide is a portion of SEQ ID NO: 7; each start position Is specified, the length of peptide Is 15 amino acids, and the end position for each peptide is the start position plus fourteen 7rJr RVTFHLPEGSQESSS 201 2701 ILHRSQAQSSV SLQQG 12 1 STTMEIWIHPQPQRK F SESTFIPGLKKMA-EI 19 110 PGLKKAAEI1P 19~ 13LTTFTP-RQAPs RG [F4 El~ PQEEYFDRTSR E18~ 153 DACWMPASLDHSSSS] 118 1278 SVSLQQGWVQGADGL ML8 291 GLCSVDQGVQGSATS 1f 1 I IRQARPSIRGDSPE 18 I lj TFEVPVSVHTRPPMK WI11 F11 KEWRSCTPMKESTT 1H7 121 RSCTPMKES1TTMEW B7 :41i.I QRKSEGKVAGKSQRR Iii 14211 RKSEGKVAGKSQRRV 17M 45]1 EGKVAGKSQRRvTFH 171 E l71 SSSDGGLGDHDAS 171 mfl AGSLTSTSHGLPLGY F17 111441 DPESTIFIPGLKKME Il 118 TfIGKM1V 171 F11 sppRVQTIALCHspp F7 g~j SPPLVQATALHHSPP Ri IILVQATALHHSPPSAQ E7] 24FJ YSPPLA kAAsHs I F27J IAQSSVSLQQGWVAQGA E1 l3Ij TSQFYTMSERLHPSD EW 13~0911 TMSERLHPSDDSIKVIE17 1]j VTTFEVPVSV TRPP E16 5i EVPVSHTRPMKEV E6 3211 MEIWIHPQPQRKSEG 16 5011 GKSQRRVTFHLPEGS L6 F5_711 TFHLPEGSQESSSDG 1E6 77 F Rn DASLSTHLPLG R~ 79 GLSSLPLGYP 16E 00 00 Table XLVI-1 09PI D4v.3- ORBi 0101-15-mers Each peptide is a portion of SEQ ID NO: 7; each start position Is specified, the length of peptide Is amino acids, and the end position for each peptide is the start position plus fourteen 182 TSTSHGLPLGYPQEE [1 187fl GLPLGYPQEEYFDRA 94, QEEYFDRATPSNRTE 1161 951 EEYFDRATPSNRTEG [1 I-h0 GDGNSDPESTFIG] FI111 STFIPGLKM1V 6 12-81 EITVQPTVEEASD-NC 16 11 NCTQECLIYGHSDAC [96 IfI-I ACWMPASLDHSSSSQ E16 FI-1 CWMPASLDHSSSSQA 16E FI111 LDHSSSSQA QASALC [6F6 IiI-11HSSSSQAQASLS n16 F16] QAQASALCHSPPLSQ- 161 FI1871 HHSPRvTQTtA LcIs 116 F-9 vTQTIALCHSPPT 6! 2-54] vTQTIALCHSPPI [16 214 PPPIQVSAU-IHSPPL 116 r2-21 U-HSPPLVQA)TALHH 16 F2261 PPLVQATALHHSP 1 6 F2331 ALHHSPPSAQASALC 6 F2--51 HHSPPSAQASACS[6 2410[SAQASALCYSPPLA 24-]1 LCYSPPLAA S F16] 2491 SPPLAQAJASHSSP J6] 2-581 ISHSSPLPQVAH 16h 268] iALHRSQAQSSVSLQ F2-12 LCSVDQGVGAS [161 300] QGsATs-Q-FYm-sERL9~ 32311 vIPLTrFTPRQQARP [9H f-7-1 PVSVHTRPPMKEWVR] 113 IRPPMKEVVRSCTM [H I i6 I MKEWVRSOTPMKESTIF6 14711 KAGKSQRRvTFHLP Hs 5611 VTFHLPEGSQESSD F15i 72 f GLGDHDAGSL [1SI 5]L 751

DHDAGSLTSTSH

85ISHGLPLGYPQ EEYFD Table XLVI-1 09P1 D4v.3- DRB1 0101-15-niers Each peptide is a portion of SEQ ID NO: 7; each start position isl specified, the length of peptideis 15 amino adds, and the end position for each peptide Is the start position plus fourteen [ij CTQECLIGSAW 1 WMPASWDHSSSQQ N AQASALCHSPPLsQA 151 R81] SQASTQHHSPRVTQTl 151 FiQ6 QHH1-sPRv-QTIALCH [15 FI98]1 L-CH-spPVTTilC 1s F21-21 HSPPPIQVSALHHSP F15 2-1-7j IQVSALHiHSPPLVA] I15~ F2- j VQATALHHSPPSQ 9 EA] AQASALCYSP=PLAQA 1~ 2651 PQVIALHRSQASV 15 N31 ERLHPSDDSIKV1PL 11-] FTable XLVII-1 09P1 D4v.3 ORBI 0301 15-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide Is 15 amino adids, and the end position for each peptide is the start position plus fourteen II1EGDGNSDPES TFIPG J EIlGLPLGYPQEEYFDRA1F24I 3FI1 DDSIKVIP-L1TFR -OI g]l EWIHPQPQRKSE-GK [9 1117 STFIPGLKKEV [9 Fl31RPPMKEVVRSCTPMK E8 E l TFHPEGSESSDG 8~ LLO] DGGLGDHDAGSLTST [98 g11 ESTFIPGLKKMEI1T E8 g12 EITVQPTVEE N [98 F2-J6 DQGVQGSAS T [98 E11 TMEIWHPQPRE [17 I 4l EGKVAGKSQRRvTFH 17- 47 KVAGKSQRRVTFHLP 7 I ]HGLPLGYPQ EEYDR 7 1104 SNRTEGDGNS-DPEST 17 Table XLVI I-i 09P1 D4v.3 DR131 0301 Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide is 15 amino acidS, and the end position for each peptide is the start position plus fourteen 12:] IPGLKKAAEITVQPT[7 264 LPQVIALH-RSQAS 7 2891 GLCSVDQGVQS 17 3261 LTTFTPRQASG 17 EVPVSVHTRPPME 16 K2]LCSVDQGVQGSATSQ6 30:]1 TSQFYTMSER5TPSD [1 781 AGSLTSTSHGLPG 144 1361 EEASDNTE Y [1 [fl KEWVRSCTPMKEST [31

SQESSSOGGLGDDAM

1g] SDGGLGDHDAST 13j E12 MA-E1VQTV ASD E13 E132QP1VEEASDNCTQEO 11M [9J kQ~v1LHRQQSSV [813 Table XLIX-T1 09P-1D4-v.3] DRB1 1101-15-mers Each peptide is a portion of SEQ ID NO: 7; each start position Is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen [j,31RPPMKEWVRSCTPMK j LPQVALHRSQAQSS g6 28!ADGLCSVDQGVQGSA E6 [IV1TEVPVSVHTRPP F2 DACWMPASLOHSSSS 22 W[EVPVSV RPME [T]MKEWRSCTPMKEST I 3jj CTPMKESTTMIH 1 3j EIWHPQPQRKSEGK EO I lj TFHLPEGSQESSSDG [8O 11-32 QPTVEEASDNCTQEc 00 00 Table XLIX-1O09P1 D4v.3 ORBi 1101-15-mersI Each peptide ii a portion of SEQ ID NO: 7; each start position Is specified, the length of peptide is amino acids, and the end position for each peptide is the start posiTionpLsfuren_ 1578 PASLDHSSSSQAA 1777 SPPLSQASTQHP 20 1931 TQTIALCHSPVQ 20 2161 PIQVSALHHSPPLVQ I nJ 265]1 PQVIALHRSQQS F20 287 jQGWVQGADGL-CS-VD 2921 LCSVDQGVQGSATSQ 320 SIKVIPLTFPRQQ 3231 VIPLTTFTPRQQRPg~ 561 VTFHLPEGSQ D [1 7721 GLGDHDAGSLTTH8 1575 CWMPASLDHSSSSQAF[1 1761WMPASLD HSSS-SQQ[ 1174 LCHSPPLSQA STQHH 8 11861 QHHSPRVTQTIALCH 6 1198 LCHSPPVTQT=IALOH F18] I24 LHHSPPLVQ=ATALHH j8 2I61 LCYSPPLAQAAAI1SH] 2511 PLAQAAAISHS=SPLP j8 28 ISHSSPLPQ\IA-LHR I6 F263 PL-PCVIALH-RSQA QS]118 F2-] ALRSQAQSSVSLQQ li 275] AQSSVSLQQWWVQGA

L

[286VQGADGLCSVDQGVQ L3-1I 1ERUIPSDDSIKVIPL F1- F1[-j EEY F DRATP S NRTE][ 189 PLGYPQEEYFDRATP 6 I fl EEYFDRATPSN=RTEG E[6 1116 STFIPGLKKAAErIT E16 ggAj CLIYGHSDACWMPAS Ill I2I4-5]ALCYSPPLAQAAAIS SQFYTMSERLIPSDD F4-fII EGKVAGKSQRRFH I 3 FW7TEVPVSVI-TRPPMK l[1B [12-9]1 STrMEIWIHPQRK B Ell1 TMEIWIHPQQRKSE F1 Table XLIX-109P1 D4v.3 DRBI 1101-15-mers Each peptide is a portion of SEQ ID NO: 7; each start position is specified, the length of peptide Is 15 amino acids, and the end position for each .peptide is the start position plus fourteen 53jj QRRVTFHLPEGSQES [Ilj DGGLGDHDAGSLTST t4] [fl] AGSLTSTSHGLPLGY F14 FJ11 STFIPGLKKMAAEITV E14 NJ MAEmIQP vE1 j4 ~JEITVQPTVEEASDNC I 14 Ii~QECLIYGHSDACW-MP 14 LIACWMPASLDHSSSSQF'4 [P1ASALCHSPPL-S6AST ,t 1-95 TLALCHSPPVTOT7IAE1 1205 TQILCSPIQ-V E14 Each peptide is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide is the start position plus eight IO9P1D4v.4-A02011 Each peptide is a portion of SEQ ID NO, 9; each start position is specified, the length of peptide is 9 amino adids, and the end position for each peptide is the start position pueiht JWPQQSQ 7.

QPQSI E J IWIHPPQS 6 109PID4v.4-Al ~I 0O9PI D4v.4 A3--rs 00 00 Each peptide is a portion of SEQ 10 NO: 9; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is t start _psition plus eight AJ IH-QPEQSQ 14 Table XXVI 1 09P1 D4v.4-A26 9-mars Each peptide is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide is 9 amino adds, and the end position for each peptide isI the start position plus eight FJWPQQSQ T F Table XXVII 109P104.4-B30702 9-mars Each peptide is a portion of SEQ ID NO: 9; each start position Is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position plus eight EJ QPQSQRRVT 1 E HFPSQR 1 N J EQllVT 221 Table XXVIII 1 09P1 D4.4-B308 9-mars Each peptide is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus. eight Al PQSQRRVTF I ]8IQSQRRVTFH IE 11-HPQPOSQ 7 Table XXIX 1 09P1 D4v.4 BI 510-9-mars Each peptide is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide is 9 amino adids, and the end position for each peptide is the start position pus e2Lt [3 I1HPQPQSQR z [jP QRVF 2 Table XXXI 109PI D4v.4 B2709-9-mers Each peptide Is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide isl 9 amino adids, and the end position for each peptide Is the start position plus eight Table XXXII 109P1 D4v.4 B4402-9-mers Each peptide is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus aight ~PQSQRRvrF Table XXXI 10911M.4-135101 9-mers Each peptide Is a portion of SEQ ID N0. 9; each start position Is specified, the length of peptide Is 9 amino adids, and the end position for each peptide is the start position plus eight QPQQRRVF 2 00 00

O

Ca Table XXXIV 109P1 4v.4-A1 10-mers Each peptide is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 3 WjHPQPOSQR 4 5~ HEQPQSQRRV 4 9 QSQRRVIFHL 4 [6 PQPQSQERVT 2~ Table XXXV 109P1D4v.4-A0201 Each peptide is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine HPQPQSQRRV 12 3 JWHPEQSR 9 QSQRRYTFHL 10 EIWIHEQPQS 7 2] IWIHPQPQSQ] 6 Table XXXVI 109P1 D4v.4- A0203 No Results Found.

Table XXXVII 109P1D4v.4 A3-10-mers 222 Each peptide is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide Is 10 amIno dds, and the end position for each peptide is the start position plus nine 3 WI11PQPQSQR 21 7 QPOSQRBVTF 1i j1 EIHPPQS 12 Table

XL-

109P1D4 v.4-B08- No Results Found.

Table XI 109P1 D4v.4- 81510 No Results Found.

Table XLII 109PID4v.4- B2705 No Results Found.

Table XLIII 109P1iD4v.4- 82709 No Results Found.

Table XLIV 109P1D4v.4-B4402 Each peptide Is a portion of SEQ ID NO- 9; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine QPQSQRRVTF E 9 QSQRRVTFHL 2 Table XXXIX 109P1D4v.4-B0702 1(-mers Each peptide is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide Is the start position pus nine 7 QPQSQRRVTF 19 5 HPQPQSQRR 17 9 QSQRRVTFHL 11 00 00 Table ML 1 09P1 D4vA4- 85101 4] T M EIW IH P QPQ0SQ RR Z 5]MEIWIHPQPQSQRRV 2] STrMEIWIHP QPQSQ 114 [NJ EIWIHPQ PQSQRRVT Hi EsTMEIWIHPQP-QS If TMEIWIHPQPQSQRg WHPQPQSQRVTH2 M9I IHPQPQSQRRVTFHL ft Table ALIX-1 09PI D4v.47 DRBi 1101-15-mers Each peptide is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide Is 15 amino acids, and the end position for each peptide is t start position plus fourteen [2]1 STTMEIWlHPQ PQSQ E2 gPQSQRRVTrFHLEGS [a []TMEIWIHPQPQS QRR E12 [MEIWIHPQPQSQ-RRV ft IHPQPQSQRRVTFHL ft-C Table XXII 1 09P1D4.5-Al 9-mers Each peptide is a portion of SEQ ID NO 1; each start position Is specified the length of peptide Is 9 amino acids, and the end position for each peptde is the start Each peptide is a portion of SEQ ID N0r 11; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start psition pus eight Table XXIV 1 09P1 m-iers Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino adids, and the end position for each peptide is the start psition plus eight 2 SVHTRESQR 24 Table XWV 9-mers Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino adids, and the end position for each peptide Is the start psition plus eight 3 SVHTRPSQR 13 N PV-SVHTRPSj1 Table XLVI l-1 09PI D4v.4 DRB1 0301.1 5-mners Each pepide is a portion of SEQ ID NO: 9; each start position Is specified, the length of peptide Is 15 amino acids, and the end position for each peptide Is the start position plus fourteen ErWIHPQPQS QRRVT 11 [4]TMEIWIPQPQCSQCRR 1 [I~jHPQPQsQRR-VTFIILP 1 [STMEIWHPQPQSQ Eia TaleXLII-109P1 D4.41 DRB1 0401-15-mers Each peptide is a portion of SEQ ID NO: 9; each start position is specified, the length of peptide Isi11 amino acids, and the end position for each peptide Is the start position plus fourteen Table XXIII I19P1D4v.51 00 00 Table XXVI 1 09P1 D4v.5-A26 9-mers Each peptide is a portion of SEQ ID NO: 11; each start position is speciie, the length of peptide is 9 amino adids, and the end position for each peptide is the start aLpRan plus eight Table XXVI I 109P1 D4v.5 830702-9-mers Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amnino acids, and the end posito for each r peptide is the star psition pius eight Table X)(VIII 1O9PID4v.5 B08-9-mers Each peptide Is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide Is the start position pus eight N RPSQRRVTF 21 109P1D4v.5 j Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 6 TRPSQRRVT n6 Table XXX IO9P1D4v.5 B32705-9-mers Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight IRPsQRRvrFl 18 4 VHTRPSQRR 14 3 SVHTRPSQR 12 8 PSQRV F 11 Table XXXI 1O9PlD4v.5 82709-9-niers Each peptide Is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino adids, and the end position for each peptide is the start psition pus eight 7] [RPSRR 13 6 T RPSQRR 11 HTRPSQRR Table XXXI I 109SP1 B4402-9-mers Each peptide is a portion of SEQ ID NO: 11; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start psition plus eight ~JL RSRRVTF 1 SVH-TRPSQR E Table XXIII I1ogP1 B5101-9-mers Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start positionlus eight 7RPSQRRVTF 13 5HTRPSQRRV I1I Table XXXIV 1O9P1D4.5-Al 1 O-mers Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide Is the start position plus nine 6] HIPQ V E2 3 VVHT E 00 00

OO

O

o Table XXXV 109P1D4v.5 A0201-10-mers Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine VHTRPAQRRV Eli Q1~sRRr]FL 6] HTRPSQRRVT 9 PSQRRTFHL 7 4] SVHTRESQRR 6 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine A PVSVHTRPSQ -Ef 4 SVHTRPSQRR IH 7ITRPSQRRVTF I

H

g HTRPSQRRV 9 9 PSQRRVTFHL 8 3 VSVHTRPSQR Table XXXIX 109P1D4v.5 80702-10-mers Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 8 RPSQRRVTFH I VPVSVHTRPS 12 6 HTRPSQRRV 11 9 PSQRRVTFHL 11 71 TRPSQRRVTFI9 .1 EI Table XLII 109P1ID4v.5 B2705-10mers No Results Found.

Table XLIII 109P1ID4v.5 B2709-10mers No Results Found.

Table XLIV 109PID4v.5-B4402 Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 7 TRPSQRRVTF 4 PSQRRVTFHL 12 Table XXXVII 109P1D4v.5-A26-10mers Table XLI 109PID4v.5 B81510-10mers No Results Found.

00 00 Table XLVI-1 09P1 D4v.5 1 DRB1 0101-15-marsI Each peptide is a portion of SEQ ID NO: 11; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteenj 1]1 VTTFEVPVSVHTRPS F D13 IRPSQRRvTFHL=PEGSF16] 7]PVSVHTRPSQRV F W41 :j0]VHTRPSQRRV F 41 D12 [TRPSQRRVTFH=LPEG M14 Table X0X11 I 09P1 D4v.6 C terminal-All 9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight j HIRPTDSRT 10 Table XXIII I 09P1 D4v.6 Cterminal-A0201 9-mers- Each peptide Is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide Is 9 amino acids, and Dhe end position for each pepfide Is the start position plus eight gj HTPTDSRT ICO I FVV 7RTI Table XXII I IO9P1D4v.6 C' terminal-A0201 9-mars Each peptide Is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide Is the start 1position plus eight ]4 VHTRPIDSRI Table XXI 10931D4v.6 0' terminal- A0203 9-mers [~ieult1 Table XXV 109P1 D4v.6 C'terminal-A3 9-mars Each peptide is a portion of SEQ ID NO: 13; each start position Is specified, the length of peptide is 9 amino acids, and the end position for eac peptie is te start position plus eight ~S1ATRPIDS 1 109PID4v.6 S Cterminal I A269-ers Table XLVIII-109P1D4.5 DRBl 0401-15-mers Each peptide is a portion of SEQ ID N(Y. 11; each start position is specified, the length of peptide is 15 amino adds, and the end position for each peptida Is the start posi!io plus fou rtee n i VrTFEVPVSVHTRPS 1122 E2 4] FEVPVSVHTRPSR FI8 i] TFEVPVSVI-TRPQ F14 8]VSVHTRPSQRRVTFH FI2 9] SVT QRTFHL F2 00 00 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 3SVHTRPTDS 11 1 PVSVHTRPT 10 11 9 HTRPTDSRT 10 2 VSVHTRPTD [5 Table XXVII 109P104v.6 C' terminal-B0702 9-mers Each pepide Is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position plus eight 1 PVSVHTRPT 10 HTRPTDSRT 9 VHTRPTDSR 4 Table XXV1II 109P1 D4v.6 C' terminal-8308 9-mers Each peptide Is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position plus eight 3SVHTRPTDS 10 HTRPTDSRT 7 Table XXIX 109P1D4v.6 C' terminal Bi510-9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus eight 4 VHTRPTDSR II 1 PVSVHTRPT 4 5 HTRPTDSRTjI Table XXX 109P1D4v.6 C' terminal-B2705 9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 4 VHTRPTDSR 12 5 HTRPTDSRT E_ Table XXXI 109P1D4v.6 C' termnal-B2709 9-mers Each peptide is a portion of SEQ ID NO: 13; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position puse ht Table XXXI 109P1 D4v.6 0' terminal-B2709 9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start ition lus eight 2 VSVHTRPTD P 5 HTRPTDSRT 2 4~ VH-TRPTDSR I Table XXXII 109PID4v.6 C' terminal-B4402 9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 3 SVHTRPTDS 4 1 PVSVHTRPT 3J 5 HTRPTDSRT 3 2 VSVHTRPTD 2J d VHTRPTDSR2 Table XXXIII 109PiD4v.6 C' terminal-B5101 9-mers 00 00 Each peptide is a portion of SEQ ID NO: 13; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start siti on plus eight ~VSVHTRPTD 4 HTRP TDSRT 2 Table XXXV IO9PID4v.6 Cterminal-Al 1 O-mers Each peptide is a portion of SEQI ID NO: 13; each start position is specified, the length of peptide Is 10 amino acids, and the end position for each peptide is the start position plus nine ]3 VVHT E 4] SVHTRPIDSR E2 Table XXXV 109P1D4v.6 C'ternmaM-0201 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide, is 10 amino acids, and the end position for each peptide Is the start psition plus nine

FZIZ

H1 SVHTRETDSR IF

T~VVVARTE

SP~vSVHIRPTD [4 jE VTRPIDSRTJP 228 FTable

XXXVI

109P1 D4v.6 A23 10-mers No Results Found.

Table XX 1I 109P1 D4v.6 Cterminal-A3 1O-mers Each peptide is a portion of SEQ ID NO: 13; each start position Is specified, the length of peptide Is 10 amnino acids, ad the end position for each peptide is the start position plus nine 1]SVHfTRPTDSR 17 [TableXXXOlII 109P1 D4v.6 Cterminal-A26 lO-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino adids, and the end position for each peptide is the start position plus nine 2] PSVHTRPTD -R1 Table XXXIX 1 09P1 D4v.6 C' terminal-B0702 Each peptide is a portion of SEQ ID NQ~ 13; each start position Is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine N Fv PV SV HTRT1 18 VIITRPTDSRTF6 Table

XL.

109P1D4 terminal- B08 [No LResults Foundj.

Table

XLI-

109P1D4 v.6-C, terminal Table

XLII-

109P1 D4 v.6-C' terminal 132705- 1lO-mers No Results Found.

10i9P1D4v.6B IC'termlnal-B ~B2709 00 00 No Results IIFound. I Table XLIV 109P1 D4v.6 C' termina-B4402 Each peptide is a portion of SEQ ID NO: 13; each start position Is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 2 PVSVHTRPTD 4 SVHTRPTDSR E3 1VPVSVHTRPT 2~ Each peptide is a portion of SEQ ID NOr 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight NDISSYVR 21HJSCLLSGTY IIMIVGFNaDI[j 7 TINCHK L8 1TCHKCLLSI8 Table XXII 109Pi1D4v.6 N' terminal-Al 9-mers TableXLVII-I 09P1 D4v.6 C' terminal-DRB1 0301 Table XXIV 109P1D4v.6 N' terminal- A0203 9-mers No Results Found.

00 00 Table XXV 109P1D4v.6 N' terminal A3-9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 14 RVT EHK 24 11 SVVRV'ITN 20 3 CLLSGIXIF 18 12 WRN"N 14 :INSISS V 33 SDISV1RVN 1 3 L1HKCLLGY 1L2 Table XXV 109P1D4v.6 N' terminal-A26 9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight I I 8 DISSWRVN 17 [IiNTTNCHKCL 1u I TTNCHKCLL 17 1I SWRVNTTN El1

MTVGFNSDI

21 HKCLLSGTY 3 2 VIGFNSDIS

[IWRVNTTNC

I SDISSWRV I1o SSWRVNTT 10 14 RVNTTNCH 10 3 CULSGTYIF W Table XXVII 109P1 D4v.6 N' terminal-80702 9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino adcids, and the end position for each peptide is the start position plus eight 9 ISSWRVNT12 16 NTTNCHKCL 10 171TNCHKCLL 10 jFNSDISSWI 9 7 SDISSWRV 9 22 KCLLSGTYI j MTVGFNSDI 8 SSWRVNTT 7 23 CLLSGTYIF jJ 22= 9 a GFNSDISSV 6 CHKCLLSGT 6 Table XXVIII 109PID4v.6 N' terminal-B08 9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 1SSRVNTT I2] 23 CLLSGTYIF 1 2 6 NTTNCHKCL 1 17TTNCHKCLL 10

TNCHKCLLS

20 CHKCLLSGT 12 WRVNTNC 8 1 MTVGFNSDIj 7 22 KCLLSGTYI 7

[I:

Table XXIX 109P1 D4v.6 N' terminal-B1i 510 9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 17 TTNCHKCLL 12 NTTNCHKCL CHKCLLSGT 9 ISSWRVNT7 CLLSGTYIF 7 DISSWRVN 6 Table XXX 109P1D4v.6 N' terminal-82705 9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 1VRVNTTNCH 14 RVNTTNCHK 23 CLLSGTYIF 6 NSDISSVVR 14 22 KCLLSGTYI 1 21 HKCLLSGTY 12 D MTVGFNSDI 11 17 TTNCHKCLL [6 NTTNCHKCL 00 00 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 4]GFNSDISSV 13 2] KCLLSGYI 2 23 CLLSGYIF 12 1 VRVNTTNCH 11 16 NTTNCHKCL 11 17 TTNCHKCLL 10 I MTVGFNSDI [s FNSDISSW Table XXXII 1 09P1D4v.6 N' terminal B4402-9-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino adcids, and the end position for each peptide is the start position plus eight 6 NTTNCHKCL 2 HKCLLSGTY 12 [23 CLLSGTYIF 12 1 TTNCHKCLL I1 22 KCLLSGTY1 I f1 MTVGFNSDI Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight [2 KCLLSGYI I MTVGFNSDI 13 SFNSDISSW 1_3 1SDISSWRV3 DISSWRVN 12 3 VGFNSDISS E GFNSDISSV 9 1NTTNCHKCL 8_ Table XXXIV 109P1ID4v.6 N' terminal-Al 10-mers Each peptide Is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine [6 NDISSYVRV I 20 CiKCLLsGTY I7 1TNCHKQLLSt4 21 NITNCHIECLL 8] Table XXXV 109P14v.6 N' terminal-A0201 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 3 VGFNSDISSV ~INSDISWRV 116 23 CLLSG.YIFA 16 8 DISSVVRVNT 13 iI ISsvEVNTrT 16 NTNc KCLL3 [4 GFNSDISSW 12 VNTTNGHKCL 9 [INCHKCLLSGT 9 Table XXXVI 109PID4v.6 N' terminal-AO203 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine 3OLSGTIFA 00 00 Table XXXVII 109PID4v.6 N' terminal-A3 1 -mers Each peptide is a portion of SEQ ID NO: 13; each start position Is specfied, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine L] KLSiY IF r1C Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine [fl OISSVVRVNT Fi [flNSDISSWRV 10 fl ISSWRVNTT Ij g15VNTNCHKCLj 16 NTINCHKCL-Li1 M SGII If ~jKCLSG1YIF 4j IGFNSISSW

I

fiA NCHKOLLSGT F[] 211HKCLSG1YI 7 23CLLSGTYIFA 7j~ VGFSDISSV Table

XL-

109P1 04 v.6 N' terminal- B08 I -mers No Results Found.

Table

XLI-

I109P1 D4 v.6 N' terminal B1510l-mers No Results Found.

Table

XLII-

109P1D4 v.6 N' termInal 82705- No Results Found.

Table

XLIII-

I09P104 v.6 N' terminal- B2709 lO-mers No Results Found.

Table XLIV-109P1 D4 v.6 N' terminal 4402-10-mers Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide is the start sition lus nine [2KCLLSG1YIF H[NTTNCHKCLLjH 20 CHCLSGY H 21HKCLLSGTYI II9 7 SDISSWRVNI jn Table

XLV-

109P1 D4 v.6 N' terminal B5101lO-mers No Results Found.

Table L(1-1a0D4V.

trmina-DR131 0101 Nte'15-mers Table XXXIX 109P1D4v.6 N' terminal-0702 1 0-mers 00 00 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide Is 15 amino acids, and the end position fort each peptide Is the start position plus fourteen I 191 NCHKCLLSG1TYIFAVI1K61 :2Ij1vGFNSDISSWRVN II ISSWRVN1TCK g2 I]SSWVRVN1TNCHC gf6 2O I CHKCLLSGTYIFAVL 16 2]1 HKCLLSGTYIFAVLL I 16 :22] KCLLSG1YIFAV 16 :18] ThCHKCLLSGTYF gi jjNSISSWRVNTTNC Table XLVI I-1 09P1 D4v.6 N' terrminal-ORBI 0301 Each peptide is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide Is 15 amino acids, and the end position for each peptide is the start position plus fourteen

TVGFNSDISSVVRVN

NSDISSWRVN1TNC jg [i111RVNTTNCHKCLG F16 E211 HKCLLSG1YIFAVlL ISSVVRVN1TNCHKC F12 [gI1SSWRVNTTNCHKCLIR2 g21 CHKCLLSG1YIFAVL I2 g1lvvRvNTrNCHKCLLS Ell @22 KCLLSG1YIFA EL El~ TNCHKCLLSG TYIFA E10 Table XLVIII-1 09P1 D4v.6 N' termlnal-DRB31 0401 Each peptide Is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 15 amino acids, and the end position for each peptide is the start position plus fourteen Table XLIX-1 09P1 D4v.6 N' terminal-ORBI 1101 15-mers Each peptide Is a portion of SEQ ID NO: 13; each start position is specified, the length of peptide is 15 amino adids, and the end position for~ each peptide is the start position plus fourteen :6]NSDISSWRNTN E2 ~j ISSWRVNTTNCHKC 1-2 :2j HKCLLSG1IFAVLL F1f2 2j 1VGFNSDISSVV Ell :I4I RVNTVNCHKCLLSGT ff Table XXII- 109P1D4v.7 N' terminalAl U-mers Each peptide is a portion of SEQ ID NO: 15; each start position Is specified, the length of peptide is 9 aino acids, and the end position for each peptide Is the start psition plus; eight LiZ llR Table XXII- 109P1 D4v.7 N' terminal-Al 9-niers Each peptide Is a portion of SEQ ID NO: 15; each start position Is specified, the length of peptide is 9 amino adids, and the end position for each peptide is the start position plus eght ENi ERVGFII 11 1O9P1D4v.7 N' terrminal-A0201 9-mers Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptde is the start position plus eight Ji~ LLVSVR E3 El E24~SL I 16 SPLLVS 19 Table XXIV- 109P1 D4v.7 N' terminal- A0203 L9-mers No Results SFound.I 00 00 Table XXV 109P1 D4v.7 N' terminal-A3 9-mers Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 17 LVVR r- 14 SLSLLLVS21 jM FLIS SS 19 F71 RVFIS1 18 LLLVSWRv 16 19 ELSRN15 Table XX" 109P1 D4v.7 N' terminai-A26 9-mers Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino adids, and the end position for each peptide Is the start! position plus eight 12 ESLPL 10 Table XXVI I 109P1 D4v.7 N' terminal-B30702 9-mers Each peptide is a portion of SEQ ID NO: 15; each start position Is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 16 SPLLLV-ySW 18 [g SSSLSPLLL 14 10 SSSSSLSPL 13 i F R VG F L 11 Table XXVIII I 09P1 D4v.7 N' terminal-B308 9-mers Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight :7J LIISSSSSL 14 11 SSSSPL1 16 SLLVW 1 14 SSLLV 17PLVSV 6 FLISSSS Table XXVIiI 1 09P1 D4v.7 N' terminal-B308 9-mers Each peptide Is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight Table XXIX 1 09P1 D4v.7 N' terminal-BiSIO1 9-mers Each peptide Is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus eight 10 SSSPL ~v.7 Each peptide Is a portion of SEQ ID NO, 15; each start position Is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position plus eight 17PLLSR t HI LISSSS 00 00 Table XXX 109P1 D4v.7 N' terminal-B2705 9-mers Each peptide Is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position plus eight :2]FRVGFLIIS 15 S S SLPL 13 11 SL SL 13 112 SSSSPLILL13 ]VGFLIISSS Table XXXI 109P1D4v.7 N' terrninal-832709 9-iners Each peptide Is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 18 LLLLVV1 LIL12 SSS 1 13 SSPLLLV w12 El RVFIS 1 SSSLSL 1 Table XXXI 109P1 D4v.7 N' termlnal-B4402 9-mers Each peptide Is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position plus ei ht R-1SLSPLLL 16 10 SSSSSLSPLH Table XXXIII 109P1D4v.7 N' terminal-B3511011 9-mars; Each peptide Is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 9 amino acids, and the end position for each poptide is the start position plus eight 18 LLSWV 1 LSPL LVS 3 00 00 Table XXXVII 1 09P1 04v.7 N' termlnal-A3 lO-mers Each peptide Is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide Is the start position plus nine 14F L-SPLLLvSVl 20 RZVFLISSS 1

LI~S

i16] §SPLLLYVWR I T8]LL1,VS-WMRVN l 14 :13]SSJLSPLLLVS Table XXXVIII 109131 D4v.7 N' terminal A26-1 0-mers Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide Is 10 amino acids, and the end position for each peptide is the start position plus nine RVGFLIISSS lE6 FI-ss~s-s~LL 14 11 SSSLS LL 1 El RGFIS E E] EISSSL No ResultsI LFound.

Table XLIII 109P1 D4v.7 N' terminal- B2709 1 O-ners No Results Found.

Table XLIV I 09P1 D4v.7 N' terminal-B34402 Each peptide Is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide Is 10 amino acids, and the end position for each peptide is the start position plus nine [HjISSSSSLSPL] 1 Table XL 109P1D4v.7 N' terminal- B08 1 0-mers Table XLI 1 09P1 D4v.7 N' termInal- B1 510 1IG-mers No Results Found.

Tabe ILI 1109P1 D4v.71 N' terminal- 1B27051 Table XLVI1 09P1 D4v.7 N' terminal-DRBI1 0101 Each peptide is a portion of SEQ ID NO: 15; each start position Is specified, the length of peptide is 15 amino adids, and the end position for each peptide Is the start position plus _fourteen 73RVGIISSLSP3 00 C0 00 Table XLVI-1 09P1 4v.7 N' terminal-ORBI 0101 Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino adds, and the end position for each peptide Is the start position plus fourteen jjj MFRVGFLIISSSSSL 5 4]1VGFIISSSSSLSPL2 211 SSSLSPLLLVSVVRV E

LSPLLLVSWRVNTTIL

5]1 GFLIISSSLSPLL g 6]j FLIISSSSSLSPLLL I[27 ISSSSSLSPLLLVSV I2I E]LVSRVN1TNCHKC E 2]j FRVGFLIISSSSs 1i El SSLSPLLLVSWRVN E XLVIII-109P1D4v.7 N' terminal-ORBI 0401 15-mers Each peptide is a portion of SEQ ID NO: 15; each start position is specified, the length of peptide is 15 amino adds, and the end position for each peptide is the start posItion plus fourteen RVGFLIISSSSSLSP R28 PLLLVSVVR C I I i] MFRVGFLIISSSSSL [)i IO] GFIISSSSSLSPL 120 j GFLIISSSSSLSPL j7 SSSLSPLLLVSWRV Ij :15] LSPLLLVSwRVNT :18LLLVS RVN NCH O 20] LVSWRVNTTNC-HKC E2O W FRVGFLIISSSSLS l 8 E j FLIISSSSSLSPL 4 16 SPLLLVSWRVNT F, 4 2~ SRNNC-HKC LIE[4 Table XXII 109P1D4v.8-AI 9-mers Each peptide is a portion of SEQ ID NO: 17; each start position Is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position plus eight OK~IT2PT 11EI Table XXIII 109P1D4v.8 A0201-9-mers Each peptide Is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight FIPGU(KEI2 Table XXIV 109PID4v.8 A0203-9mers III9PI D4v.8I 00 00 a

CL)

Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide Is the start position lus eight GLNKE!IVQ 16 8 KETVQRTV 11 2 FlEGLK EI 10] SLK EI QP 9E i lTFPGLKKE 8- Table XXVI 109P1D4v.8 A26-9-mers Each peptide Is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino adcids, and the end position for each peptide is the start psition plus eight 1 1 TFIPGLKKE 11 SFIPGLKKEl EI H I= 6 LXKEITVQP 5 aJIKEITVQPTV 5 Table XXVII 109P1D4v.8 B0702-9-mers Each peptide Is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 3 IPGLKKEIT 8 239 Table XXVII 109PiD4v.8 B0702-9-mers Each peptide is a portion of SEQ ID NO. 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 7 KKEIWVQPT Table XXVIII 109P1D4v.8 B08-9-mers Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position lus eight SGLKKEITVQ 18 2 FIPGLKKEI 13 6 LKKEITVQP 13 4 PGLKKEITV 10 Table XXIX 109P1D4v.8 81510-9-mers Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 5 GLKKEITVQ Table XXIX 1 09P1 D4v.8 B1510-9-mers Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 1~ TFIPGLKKE 4 2 FIPGLKK(EI 3 IPGLKKEIT 3 6 LKKEITVQP Table XXX 1 9PI D4v.8 B2705-9-mers Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide is the start position plus eight 5 GLKKEI1TVQ [g 2 FIPGLKKEI 11 8 KEITVQPTV 9 1 TFPGLKKE Table XXXI 109P1D4v.8 B2709-9mers 00 00 Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position plus eight 8]K EI1TV QPTv 12 21 FI PGLK-K-E-I [E8 Table)XXXII IO9PID4v.8 B4402-9-mers Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 9 amino acids, and the end position for each peptide is the start position pus eight 2J FIPGLK-KEI 12 Table XXXIII 1 09P1 D4v.8 85101-9-niers Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide Is 9 amino acids, and the end position for each peptide Is the start position plus eight PGLKK E 21 21 FIGK E14 IGKET13 1 KI1Q1V 1 z39J Table XXXIV 1 09P1 D4v.8 Al-i 0-mers Each peptide is a portion of SEQ ID NO: 17; each start position Is specified, the length of peptide isl10 amino ads, and h end position for each peptide is the start position pus nine JSFIP!J7KE 1 Table XXX 109P1D4v.8 A0201-1O-er Each peptide is a portion of SEQ ID NO: 17; each start position Is specified, the length of peptide is 10 amino acids, and the end position for each peptide is the start position plus nine qJ FIPGL1KEI-T i51 I PGL-K1EI1V 14 q1 TFIPGIKE13 IOj KKET-YQPTV i STFIP GLKKE 12 ffl LKKEIIVQPT 1 Table 109P1 D4v.8 A0203-1O0mers Table XXII fl I9P1D4v.8 A3-10-ers- Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amnino acids, and the end position for each peptide is the start position plus nine GL KKEIW-VQ-P 1 §KEITV~ifTVE 1 Table XXXIII 1 09P1 D4v.8 A26-1O-mers Each peptide Is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide Is amino adids, and the end position for each peptide is the star position plus nine Table XXXI 1 09PI D4v.8 B0702-1O-mers Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino adds, and the end position for each peptide Is the start position plus nine H IPGLKKEI1TV[8 ELKKEITrVQPT Table XL IO9PID4v.8 B08-1 0mers 00 00 Table XiJ 109P1 D4v.8 B151O-1Omers 10O9Pl D4v.8j B82705-1 0-I No Results Found.

Table XLII I 1 09P1 D4v.8 B2709-1 0mers Table XLIV 1 091Dv.8 B4402-10-mers Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide is 10 amino acids, and the end position for each peptide Is the start position plus nine 2]KITVPTE61 ETable XLV 109P1 D4v.8 B5101-%0 Mers No Results 1Found.

Table XLVI-1 09P1 D4v.8 OR~I3 0101-15-mers Each peptide is a portion of -SEQ ID NO: 17; each start position Is specified, the length of peptide Is 15 amino acids, and the end position for each peptide Is the start position pius fourteen 9]1 IPGU(KEI1rvopT1v-El P25 ij3] KKEITVQPT VEEASD E21 5]1 ESTFIPGL EIV Jj9 3]1 DPESTFIPGLKKEIT IiE ]J STFIPGUEIVP1 i:2J LKKEITVQPTVEEASJ Table XLVI I-I 09P1 D4v.81 DRB1 0301-15-mers Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptide Is 15 amino acids, and the end position for each peptide is the start position plus fourteen 51 ESTFIPGLKE17 61 STFIPGLKKEI1 7I 9]1 IPGU<KEITVQPTVE [12 NSDPES TFIPGU(KKE Table XLVIII-1 09P1 D4v.8 DRBI 0401-15-mers Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of pepide Is 15 amino adids, and the end position for each peptide Is the start position plus fourteen :61STFIPGLKKEI1TVQPI E2 :9]1 PGuKKITwQPiE 2C 5]1 ESTFIPGLKKEiVQ0 16 :13][KKEITVQPVE E14 :2][SDPEsTFIPGuoKE I12 :3]1DPESTFIPGKEr 12 i0]j PGLKKEI[TVQP1VEE 2 i11] GL-KKEIMVP1VEEA [t2 Table XLIX-109P1 D4v.81 DRB1 1101-15-mersJ Each peptide is a portion of SEQ ID NO: 17; each start position is specified, the length of peptlde is amino adds, and the end position for each peptide is the start position plus fourteen 00 00 Table L: Protein Characteristics of 109P11)4 109PID4 var.1 Bioinformatic URL on World Wide Web Outcome Program ORF ORF finder Protein length Transmembrane region TM Pred .ch.ctnbnet~org/ HMMTop .enzin-Lhu/hnuntop/ Sosui .genome.adjp/SOSui/ TMHMM .cbs.dtu.dktservicewiTMHMM Signal Peptide Pi Molecular weight Localization Motifs Signal P p1/MW tool p1/MW tool

PSORT

PSORT 11 Pfam Prints Blocks .cbs.dtu.dklservices/SignalP/ .expasy.ch/tools/ .expasy.chltooWs psortnibb.ac.jp/ psottnibb.ac.jp/ .sanger.ac.uklPfaml .biochezn~ucl.ac.ukl .blocks.flxrc.org/ 846-3911 bp (includes stop codon) 1 021 an 3 TM helices (aa3-aa23, aa756aa776, aa8l 6-aa834), N terminus intracellular no TM, N terminus extracellular 3 TM helices (2-24aa, 756-778aa, 81 O-832aa), N terminus extracellular I TM helix (813-835aa), N terminus extracellular yes pI 4.81 112.7 kDa Plasma membrane 67% endoplasmic reticulum Cadherin domain Cadherin domain, DNA topoiso- Merase 4B, sonic hedgehog Cadherin domain, uibosomal protein Ll I0E, ribulose biphosphatc carboxylase (large chain), ornithine decarboxylase antizyme protein phosphatase 2C subfamily Table U. Exon boundaries of transcript 109Pi D4 v.1 Table 1.1I(a). Nucleotide sequence of transcript variant iO09PI D4 v.2 (SEQ ID NO: 237) cccctttctc cccctcggtt aagtccctcc ccctcgccat; tcaaaagggc ctggctcctt gcagtcggcg aactgtcggg gcgggaggag ccgtgagcagI cagctgcccg cgcggcaaag aggaaggcaa gccaaacaga gtgcgcagagI agcggcgaca caggcagcac aggcagcccg ggctgcctga atagcctcag agcgactccg gctgctctgc ggactgcgag ctgtggcggt agagcccgct.

cagtctccgt ggagcgggcg gaagcctttt ttctcccttt cgtttacctcI tctaaaggca tcgttattag gaaaatcctg ttgcgaataa gaaggattcc taccggagag gttttgcctc agctgctctc aactttgtaa tcttgtgaag gcttggctga ttgcagagca ctatgaggac tgaacgacag tgggttttaa tcaagtgttg tgcgggttaa tacaacaaac tgtaacaagt gtacctggta gtccgggacg tacattttcg cggtcctgct agcatgcgtg gtgttccact ggagaaaaac tacaccatcc gagaagaaat gccagaaaac gtcctgatag gaaagacctt aacttgtcgc tgattccaaa caagtccttg acaactgcta gctagtgtac aagaccggag atgtgccact gattcgaatt gaagaggata cttcactact ggcgctcgca ttgatcgtga gaaattatgt gctggtatcc gcattgcttt tatgaagtgg aggttgccat tttgccggat gaaatattta gatacgtttt ctgatagaag atataaatga taatgcacca ttgttcccag caacatatca attccagaga actcggctat aaactctaaa tatactctc tgatcctgac gtaggaataa acggagttca aaactacgaa ctaattaaga ttttggcctc gatgtcattg aaacaccaga aggagacaag atgccacaac aaagqaqtta gataqggaag agaaggatac ctacgtgatg aaagtaaagg :ggctcggca :agctgcact :ggcagtgcc aaacaacctc acagcagtcg ttcattctac acagatcaca aagctgacaa Ltcagatatt tggacttgtt ctggcgccCa gcgacttgtt tgcagttcaa ctggtgagat caagggatga gactggttaa caacagttat cagcggctgt gtcaaaacat tgattgttca ttgaagatgg 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 00 00 tggctttcct caaccaccca aggcacttca Ccacttctct caccactgga gttactggtt tgttacagat caatgacaca tgtgacggat ccct tt caga tgactatgag tcctttqaat agttttcacc gttgacgaaa gctaggccct agtgaagaaa taacggggta tgacaatagc aaggcatggt agttacgctc catccgacca ggctgaggat tgatgtcaat ggttctaccg cactggcatg tgcaatcgac tggtttacac tgttgtaatt actggtgcgc ctcaccaact cgttgtagtt tgctcagaaa gataatgatg ttttgtcact actagacctt acctactact acagcctgcc agaactgcct cagttcagat tgtgtccgta gatataactt gatttcaaaa agcaaaaatc tggcaatgqa atgctgtttc gtatacactt gttgaataat tctctgaaag accaaaacac tcatttttat cctgtaatcc accatcctgg cgtggtggcg aacccgggag acagagcaag aggtttttat tttggctggg ggatcacctg caaagatcca qtctttaagg gtgacacagc ttcagcaatc cttatcacaa ttggcaagtg gtcaatgata gttgttCttt aagqatgcgg ttaaggccag tccacaaaag cagtcagcaa cagtctttcg gtaagtgcaa gatgctccac ctagatagag ccacccttaa ccagttttca acagtaggac tccattttag aatatttcat ggtggtagag gacaacaaac tccactaatc aatgcagagg caagaaacag agagtgttgg gtcaatctgt aaaagcactg agtgactatg attttcatca aacaagcaga aagaaaaaga attgaagaaa cctattgatc ttcaagcccg ttccaaattc ctcgataaca ccctacagcg cacaccagac tctaggaaca ttaggctaag atcttaaaaa aatttaaaat tctgtttgct atggttaata taaaacatta gtatccaggg tctggttcag tacccaccac cagcactttg ctaacacggt ggcgcctgta gcggagcttg actctgtctc gttgattgta tgtggtggct aggtcaggag gtactgctat agacagagat tccatgccac tagtctccaa tcaaagaacc atggtggatt atgtcccatc cagaaaatat accataatgg tattcagtaa aatatgccat tgctcttcat taactgtttc tggatgcaga ctgaattcag aaaaagagga ccagcaatgt ctcacaatga taatcactgt atgagaatga ttgatagaga tatcacgttc cagttttcat caggcacagt ttcgttacag gcaacataac tcaaagctaa tcgtgaatga aagcaccagt tcaagatcct ctgctgtagt attctgaatg aaaagaagaa ctaaggcaga tagaagagca acagccctga agcctgaaac cctttgtggc tttctgactg cgactgattc acaaaattc atcattaatt tgatgtccta ttatqgaaga ctgaatcaac tatcagtcat tctccaggag caagagacat tgttttgaaa tctgcttaaa ggaggccgag gaaaccccat gtcccagcta cagtgagccg aaaaagaaaa ctcatgctgt cacacctgta ttcaagacca tttgcaaqtq tgaagtcagt agatqctgac cattgccagg actggatagg gatgccagca cattgacata tccactcaac cagggtgaca tcagttcctc taaattactg caaagtgaaa tattcctgag cagtgggcct cctggattgt taaatattta cacagtcttt atacaacttc aactgatcct tgacttcacc aaaacaagaa ttcaagtgcc tgtccctcct ggtctttcag cattgtagga attgatggag tgacttagga gtcggtgacc gaccccaaat ggttgcagct aagatgtcgc ggctacccca gaagcattcc tgatgttgac aacaatggga tttggcccga tcccctgaat ctgtgactct tggctatcca caggacatca attccccttc ttgtaatcta gtgaaccttg gacagtgcag agccatgatg gaaacatgca tttggaagtg ttttaagacc atattcacta.

agttgagtgg gcgggtggat ctccactaaa ctcgggaggc agatggcgcc aaatgttcaa.

tccactcctt atcccagcac gtctggccaa agtgttactg ataccagaaa ataggtgaaa agattatttc qaagaaacac agagcaatgg agatacatcg accaaaattg tgcttcacag ctggagactg gctgcagatg gatgaaaatg aataactcte aatgctaaga cgtacaggca ttcacaattc gtaagcatta tatgtcccag gattatggag attgattcac tcttacactt aaagtaacca tccaactgtt gtaattgctg ggaaacacaa aaatgtgatg cagcctgatt aatgctacac actgagatag gttgctggca caggoaccac aacccagaaa cctaagaact agtgatggaa aagtacaatt cactacaaat tcgaagcacc atctccaagt gtgacgacct actattgaaa.

caaaaaattt gatttcccat tgctttcttt cacaataaca taatataagg attacttgcc agctgaacta ccaaacaaac acataatatt gccgg9gcgg cacgaggtca aatacaaaaa tgaggcagga actgcactcc tgatagaaaa ttaattatta tttgggaggc cat atacaaatga atgctcctgt atgccaagat acctcaatgc caaaccacaa tgctggtaaa tcaatcctgt ctctcataac atcatgaaat cagcatatct ctggcaaacc acaatgctcc ctggcatcca tcaattacct tgctgactgt tggcaaaaga ttgatcagaa aaaaccttcc acaattctgc aaactggtgt tctatgtaaa taaatgtggt cttatgaatt ttgacaatga gagatctgtt ttacagacct ctctcttcag tgattaatga ctgatgtatc ccataactgt accttaaggc acaqgcagat tgctgcttaa acagagtcac gggtaactac ctgcctctcc acatcatcca gttcctcaag tcgaggtacc tctgcagtga caatgattgt tataaaagca agctgtaatc gagtactctc ctgtcttggt ctgtctgatt gccaaactac aaaaaacaaa gctgagaaaa gtggctcacg ggagattgag attagcctgg gaatagcgtg agcctgggtg taattttact aaaagttatt cgaggtgggt 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4723 00 Table UII(a). Nucleotide sequence alignment of 10911D4 v.1 (SEQ ID NO: 238) and 109PI1D4 v.2 (SEQ ID NO: 239) Score =5920 bits (3079), Expect =0.0ldentities =3079/3079 (100%) Strand =Plus!I Plus V.1 800 agtgttgtgcgggttaatacaacaaactgtaacaagtgtacctggtatggacttgttgtc 859 CAV.2 544 agtgttgtgcgggttaatacaacaaactgtaacaagtgtacctggtatggacttgttgtc 603 V.1 860 cgggacgtacattttcgcggtcctgctagcatgcgtggtgttccactctggcgcccagga 919 V.2 604 cgggacgtacattttcgcggtcctgctagcatgcgtggtgttccactctggcgcccagga 663 V.1 920 gaaaaactacaccatccgagaagaaatgccagaaaacgtcctgataggcgacttgttgaa 979 664 gaaaaactacaccatccgagaagaaatgccagaaaacgtcctgataggcgacttgttgaa 723 00V.1 980 agaccttaacttgtcgctgattccaaacaagtccttgacaactgctatgcagttcaagct 1039 V.2 724agaccttaacttgtcgctgattccaaacaagtccttgacaactgctatgcagttcaagct 783 V.1 1040 agtgtacaagaccggagatgtgccactgattcgaattgaagaggatactggtgagatctt 1099 V.2 :784 agtgtacaagaccggagatgtgccactgattcgaattgaagaggatactggtgagatctt 843 V.1 :1100 cactactggcgctcgcattgatcgtgagaaattatgtgctggtatcccaagggatgagca 1159 V.2 :844 cactactggcgctcgcattgatcgtgagaaattatgtgctggtatcccaagggatgagca 903 V.1 :1160 ttgcttttatgaagtggaggttgccattttgccggatgaaatatttagactggttaagat 1219 V.2 :904 ttgcttttatgaagtggaggttgccattttgccggatgaaatatttagactggttaagat 963 V.1 :1220 acgttttctgatagaagatataaatgataatgcaccattgttcccagcaacagttatcaa 1279 V.2 :964 acgttttctgatagaagatataaatgataatgcaccattgttcccagcaacagttatcaa 1023 V.1 :1280 catatcaattccagagaactcggctataaactctaaatatactctcccagcggctgttga 1339 V.2 :1024 catatcaattccagagaactcggctataaactctaaatatactctcccagcggctgttga 1083 V.1 :1340 tcctgacgtaggaataaacggagttcaaaactacgaactaattaagagtcaaaacatttt 1399 V.2 :1084 tcctgacgtaggaataaacggagttcaaaactacgaactaattaagagtcaaaacatttt 1143 V.1 1400 tggcctcgatgtcattgaaacaccagaaggagacaagatgccacaactgattgttcaaaa 1459 V.2 1144 tggcctcgatgtcattgaaacaccagaaggagacaagatgccacaactgattgttcaaaa 1203 V.1 1460 ggagttagatagggaagagaaggatacctacgtgatgaaagtaaaggttgaagatggtgg 1519 V.2 :1204 ggagttagatagggaagagaaggatacctacgtgatgaaagtaaaggttgaagatggtgg 1263 V.1: 1520 ctttcctcaaagatccagtactgctattttgcaagtgagtgttactgatacaaatgacaa 1579 V.2 :1264 ctttcctcaaagatccagtactgctattttgcaagtgagtgttactgatacaaatgacaa 1323 244 00 V.1 1580 ccacccagtctttaaggagacagagattgaagtcagtataccagaaaatgctcctgtagg 1639 V.2 1324 ccacccagtctttaaggagacagagattgaagtcagtataccagaaaatgctcctgtagg 1383 V. 60cctatgccgtctcaagtNgitgtaaagcaaca19 V.2 1684 cacttcagtgacacagctccatgccacagatgctgacataggtgaaaatgccaagatcca 1699 V.1 1384 cattctgcacagtctatccacaatgctcaagtgaatc caagccac 1443 V.2 1700 cttctctttcagcaatctagtctccaacattgccaggagattatttcacctcaatgccac 1593 V.1 1760 cactggacttatcacaatcaaagaaccactggatagggaagaaacaccaaaccacaagtt 1819 V.2 1504 cactggacttatcacaatcaaagaaccactggatagggaagaaacaccaaaccacaagtt 1563 00 V.1 1820 actggttttggcaagtgatggtggattgatgccagcaagagcaatggtgctggtaaatgt 1879 ciV.2 :1564 actggttttggcaagtgatggtggattgatgccagcaagagcaatggtgctggtaaatgt 1623 V.1 :1880 tacagatgtcaatgataatgtcccatccattgacataagatacatcgtcaatcctgtcaa 1939 V.2 :1624 tacagatgtcaatgataatgtcccatccattgacataagatacatcgtcaatcctgtcaa 1683 V.1 :1940 tgacacagttgttctttcagaaaatattccactcaacaccaaaattgctctcataactgt 1999 V.2 :1684 tgacacagttgttctttcagaaaatattccactcaacaccaaaattgctctcataactgt 1743 V. 1 :2000 gacggataaggatgcggaccataatggcagggtgacatgcttcacagatcatgaaatccc 2059 V.2 ;1744 gacggataaggatgcggaccataatggcagggtgacatgcttcacagatcatgaaatccc 1803 V.1 :2060 tttcagattaaggccagtattcagtaatcagttcctcctggagactgcagcatatcttga 2119 V.2 1804 tttcagattaaggccagtattcagtaatcagttcctcctggagactgcagcatatcttga 1863 V.1 2120 ctatgagtccacaaaagaatatgccattaaattactggctgcagatgctggcaaacctcc 2179 V.2 1864 ctatgagtccacaaaagaatatgccattaaattactggctgcagatgctggcaaacctcc 1923 V.1 2180 tttgaatcagtcagcaatgctcttcatcaaagtgaaagatgaaaatgacaatgctccagt 2239 V.2 1924 tttgaatcagtcagcaatgctcttcatcaaagtgaaagatgaaaatgacaatgctccagt 1983 V.1 2240 tttcacccagtctttcgtaactgtttctattcctgagaataactctcctggcatccagtt 2299 V.2 1984 tttcacccagtctttcgtaactgtttctattcctgagaataactctcctggcatccagtt 2043 V.1 2300 gacgaaagtaagtgcaatggatgcagacagtgggcctaatgctaagatcaattacctgct 2359 V.2 2044 gacgaaagtaagtgcaatggatgcagacagtgggcctaatgctaagatcaattacctgct 2103 V.1 2360 aggccctgatgctccacctgaattcagcctggattgtcgtacaggcatgctgactgtagt 2419 V.2 2104 aggccctgatgctccacctgaattcagcctggattgtcgtacaggcatgctgactgtagt 2163 00 V.1 2420 gaagaaactagatagagaaaaagaggataaatatttattcacaattctggcaaaagataa 2479 V.2 2164 gaagaaactagatagagaaaaagaggataaatatttattcacaattctggcaaaagataa 2223 V.1 2480 cggggtaccacccttaaccagcaatgtcacagtctttgtaagcattattgatcagaatga 2539 V.2 2224 cggggtaccacccttaaccagcaatgtcacagtctttgtaagcattattgatcagaatga 2283 V.1 2540 caatagcccagttttcactcacaatgaatacaacttctatgtcccagaaaaccttccaag 2599 V.2 2284 caatagcccagttttcactcacaatgaatacaacttctatgtcccagaaaaccttccaag 2343 CN1 V.1 2600 gcatggtacagtaggactaatcactgtaactgatcctgattatggagacaattctgcagt 2659 CN1 V.2 2344 gcatggtacagtaggactaatcactgtaactgatcctgattatggagacaattCtgcagt 2403 00 V.1 2660 tacgctctccattttagatgagaatgatgacttcaccattgattcacaaactggtgtcat 2719 V.2 2404 tacgctctccattttagatgagaatgatgacttcaccattgattcacaaactggtgtcat 2463 V.1 2720 ccgaccaaatatttcatttgatagagaaaaacaagaatcttacactttctatgtaaaggc 2779 V.2 2464 ccgaccaaatatttcatttgatagagaaaaacaagaatcttacactttctatgtaaaggc 2523 V.1 2780 tgaggatggtggtagagtatcacgttcttcaagtgccaaagtaaccataaatgtggttga 2839 V.2 2524 tgaggatggtggtagagtatcacgttcttcaagtgccaaagtaaccataaatgtggttga 2583 V.1 2840 tgtcaatgacaacaaaccagttttcattgtccctccttccaactgttcttatgaattggt 2899 V.2: 2584 tgtcaatgacaacaaaccagttttcattgtccctccttccaactgttcttatgaattggt 2643 V.1 :2900 tctaccgtccactaatccaggcacagtggtctttcaggtaattgctgttgacaatgacac 2959 V.2: 2644 tctaccgtccactaatccaggcacagtggtctttcaggtaattgctgttgacaatgacac 2703 V.1 :2960 tggcatgaatgcagaggttcgttacagcattgtaggaggaaacacaagagatctgtttgc 3019 V.2: 2704 tggcatgaatgcagaggttcgttacagcattgtaggaggaaacacaagagatctgtttgc 2763 V.1 :3020 aatcgaccaagaaacaggcaacataacattgatggagaaatgtgatgttacagaccttgg 3079 V.2 :2764 aatcgaccaagaaacaggcaacataacattgatggagaaatgtgatgttacagaccttgg 2823 V.1 :3080 tttacacagagtgttggtcaaagctaatgacttaggacagcctgattctctcttcagtgt 3139 V.2 :2824 tttacacagagtgttggtcaaagctaatgacttaggacagcctgattctctcttcagtgt 2883 V.1 3140 tgtaattgtcaatctgttcgtgaatgagtcggtgaccaatgctacactgattaatgaact 3199 V.2 2884 tgtaattgtcaatctgttcgtgaatgagtcggtgaccaatgctacactgattaatgaact 2943 V.1 :3200 ggtgcgcaaaagcactgaagcaccagtgaccccaaatactgagatagctgatgtatcctc 3259 V.2 :2944 ggtgcgcaaaagcactgaagcaccagtgaccccaaatactgagatagctgatgtatcctc 3003 246 00 V.1 3260 accaactagtgactatgtcaagatcctggttgcagctgttgctggcaccataactgtcgt 3319 V.2 3004 accaactagtgactatgtcaagatcctggttgcagctgttgctggcaccataactgtcgt 3063 V.1 3320 tgtagttattttcatcactgctgtagtaagatgtcgccaggcaccacaccttaaggctgc 3379 V.2 3064 tgtagttattttcatcactgctgtagtaagatgtcgccaggcaccacaccttaaggctgc 3123 V.1 3380 tcagaaaaacaagcagaattctgaatgggctaccccaaacccagaaaacaggcagatgat 3439 V.2 3124 tcagaaaaacaagcagaattctgaatgggctaccccaaacccagaaaacaggcagatgat 3183 V. 40atagagaagaagaagagatccagattcgtatt39 00V.2 314 aatgatgaagaaaaagaaaaagaagaagaagcattcccctaagaacttgctgcttaattt 3249 00V.2 3184 atgataagaaaagaaaaagacagaagaagttcctaaggaacgctcacatt 3243 .NiV.2 3500 tgtcactattgaagaaactaaggcagatgatgttgacagtgatggaaacagagtcacact 3359 V.1 3560 agaccttcctattgatctagaagagcaaacaatgggaaagtacaattgggtaactacacc 3619 V.2 3304 agaccttcctattgatctagaagagcaaacaatgggaaagtacaattgggtaactacacc 3363 V.1 3620 tactactttcaagcccgacagccctgatttggcccgacactacaaatctgcctctccaca 3679 V.2 3364 tactactttcaagcccgacagccctgatttggcccgacactacaaatctgcctctccaca 3423 V.1: 3680 gcctgccttccaaattcagcctgaaactcccctgaattcgaagcaccacatcatccaaga 3739 V.2 :3424 gcctgccttccaaattcagcctgaaactcccctgaattcgaagcaccacatcatccaaga 3483 V.1: 3740 actgcctctcgataacacctttgtggcctgtgactctatctccaagtgttcctcaagcag 3799 V.2. 3484 actgcctctcgataacacctttgtggcctgtgactctatctccaagtgttcctcaagcag 3543 V.1 ;3800 ttcagatccctacagcgtttctgactgtggctatccagtgacgaccttcgaggtacctgt 3859 V.2 :3544 ttcagatccctacagcgtttctgactgtggctatccagtgacgaccttcgaggtacctgt 3603 V.1 :3860 gtccgtacacaccagaccg 3878 V.2 :3604 gtccgtacacaccagaccg 3622 Table UV(a). Peptide sequences of protein coded by 1 09P1 D4 v.2 (SEQ ID NO: 240) MRTERQWVLI QIFQVLCGLI QQTVTSVPGM DLLSGTYIFA VLLACVVFHS GAQEKNYTIR EEMPENVLIG DLLKDLNLSL IPNKSLTTAM QFKLVYKTGD VPLIRIEEDT GEIFTTGARI 120 DREKLCAGIP RDEHCFYEVE VAILPDEIFR LVKIRFLIED INDNAPLFPA TVINISIPEN 180 SAINSKYTLP AAVDPDVGIN GVQNYELIKS QNIFGLDVIE TPEGDKMPQL IVQKELDREE 240 KDTYVMKVKV EDGGFPQRSS TAILQVSVTD TNDNHPVFKE TEIEVSIPEN APVGTSVTQL 300 HATDADIGEN AKIHFSFSNL VSNIARRLFH LNATTGLITI KEPLDREETP NHKLLVLASD 360 GGLMPARAHV LVNVTDVNDN VPSIDIRYIV NPVNDTVVLS ENIPLNTKIA LITVTDKDAD 420 HNGRVTCFTD HEIPFRLRPV FSNQFLLETA AYLDYESTKE YAIKLLAADA GKPPLNQSAM 480 LFIKVKDENIJ NAPVFTQSFV TVSIPENNSP GIQLTKVSAM DADSGPNAKI NYLLGPDAPP 540 EFSLDCRTGM LTVVKKLDRE KEDKYLFTIL AKDNGVPPLT SNVTVFVSII DQNDNSPVFT 600 00HNEYNFYVPE NLPRHGTVGL ITVTDPDYGD NSAVTLSILD ENDDFTIDSQ TGVIRPNISF 660 DREKQESYTF YVKAEDGGRV SRSSSAKVTI NVVDVNDNKP VFIVPPSNCS YELVLPSTNP 720 GTVVFQVIAV DNDTGMNAEV RYSIVGGNTR DLFAIDQETG NITLMEKCDV TDLGLHRVV 780 KANDLGQPDS LFSVVIVNLF VNESVTNATL INELVRKSTE APVTPNTEIA DVSSPTSDYV 840 KILVAAVAGT ITVVVVIFIT AVVRCRQAPH LKAAQKNKQN SEWATPNPEN RQMIMMKKKK 900 KKKKHSPKNL LLNFVTIEET KADDVDSDGN RVTLDLPIDL EEQTMGKYNW VTTPTTFKPD 960 SPDLARHYKS ASPQPAFQIQ PETPLNSKHH IIQELPLDNT FVACDSISKC SSSSSDPYSV 1020 SOCGYPVTTF EVPVSVHTRP TDSRTSTIEI CSEI 1054 Table LV(a). Amino acid sequence alignment of 109P1D4 v.1 (SEQ ID NO: 241) and 109P1D4 v.2 (SEQ ID NO: 242) Score =2006 bits (5197), Expect =0.Oldentities =1012/1017 Positives =1013/1017 (99%) V.1 1 MDLLSGTYIFAVLLACVVFHSGAQEKNYTIREEMPENVLIGDLLKDLNLSLIPNKSLTTA MDLLSGTYI FAVLLACWVHSGAQEKNYTIREEMPENVLIGDLLKDLNLSLIPNKSLTTA V.2 30 MDLLSGTYIFAVLLACVVFHSGAQEKNYTIREEMPENVLIGDLLKDLNLSLIPNKSLTTA 89 00V.1 61 MQFKLVYKTGDVPLIRIEEDTGEIF'rTGARIDREKLCAGIPRDEHCFYEVEVAILPDEIF 120 0Q0VKGVLREDGITGRIRKCGpDR~EEA~DI V.2 90 MQFKLVYKTGDVPLIRIEEDTGEIFrTGARIDR.EKLCAGIPRDEHCFYEVEVAILPDEIF14 (iV.2 121 MQLVKJIDIPLFPAEETVEINISPNAISKLAAPRDVGINGEVVQNPEIK 180 V.1 121RLVKIRFLIEDINDNAPLFPATVINISI PENSAINSKYTLPAAVDPDVGINGVQNYELIK 18 V.2 50 V.1 181 SQNIFLIETPEGDAPLPTIIVQELREKDSyVIjSKLAVPDGFQSINGVQV 2409 V.1 181SQNIFGLDVIETPEGDKMPQLIVQKELDREEKDTYVMKVKVEDGGFPQRSSTAILQVSVT24 V.2 210SQNIFGLDVIETPEGDKMPQLIVQKELDREEKDTYVMKVKCVEDGGFPQRSSTAILQVSVT26 V.1 210 DTNNHPLVETEEVSIPNA2VGTSVLEKTADIGEDGGIFFSLSTAILP 300 V.1 241DTNDNHPVFKETEIEVSI V.2 270DTNDNHPVFKETEIEVSIPENAPVGTSVTQLHATDADIGENAKiIHFSFSNLVSNIA.RLF32 V.1 370 HLNATTGLIKEPLDEIEPHKLLVLDGGI.IPAPNLVNV'VNDVSIIRYI 360 V.1 301HLNATTGLIT IKE PLDREETPNHKLLVLASDGGLMPARAMVLVNVTDVN DNVPSIDIRYI36 V.2 330HLNATTGLITIKEPLDREETPNHKLLVLASDGGLPARAVLVNqVTDVNDNVPSIDIRYI38 V.1 361 VNPNTVLSIENPLDPNTPNHALTVKADHGMPRTCFI'DHEIPFRLRPSQFLLE 420 V.1 361VNPVNDTVVLSENIPLNTKIALITVTDKDADHiNGRVTCFTDHEIPFRLRPVFSNQFLLET42 V.2 390VNPVNDTVVLSENIPLNTKIALITVTDKDADHN'GRVTCFTDHEI PFRLRPVFSNQFLLET49 V.2 421 AAYLNDYEVSTEYIKLLAADAGKPPLNQSAI4LFIKVTKDENPFtQSFSIPFET 480 V.1 421AAYLDYESTK(EYAIKLLAADAGKPPLNQSAMLFIKVKDENDNAPVFTQSFVTVSI PEWNS48 V.2 450AAYLDYESTKEYAIKLLAADAGKPPLNQSAMLFIKVKDENDNAPVFTQSFVTVSI V.1 481 PGIOLDTKAI AJJAK LPDAPEFSLVEDCRTMLVKXLDREKEDKYTI 509 V.1 481PGIQLTKVSAIDADSGPNAKINYLLGPDAPPEFSLDCRTGMLTVVKKLDREKEDKYLErI54 V.2 510PGIQLTKVSAMDADSGPNAKINYLLGPDAPPEFSLDCRTGMLTWVKXLDREKEDKYLFTI56 V.1 510 LAXDGPLTK SNVTVIVSIIDOKNLDSP~H PE NLPRIIGTLT VLITDPYG~ 560 V.1 541LAKDNGVPPLTSNVTVFVSIIDQNDNSPVErHNEYNFYVPENLPRHGTVGLITVT1DPDYG60 V.2 570LAKDNGVPPLTSNVTVFVSIIDQN4DNSPVErHNEYNFYVPENLPRHGTVGLITVTDPDYG62 V.1 6 50 LDNAVPLSEVIIJ IDSQNRNSFTNEYFYVAEDRGTGLVSSSSAK 660 V.1 601DNSAVTLSILDENDDFTDS'IGVIRPNISFDREKQESYTFYVKAEDGGRVSRSSSAKVT60 V.2 630DNSAVTLSILDENDDFTIDSQIGVIRPNISFDREKQEM FYVKAEDGGRVSRSSSAKVT68 V.1 610 INSVDV DNVFIVPSCSYVLRPNSTNPGTVVQVIAV DGAVRSVT 720 V.1 661INVVDVNDNKPVFIVPPSNCSYELVLPSTNPGTVVFQVIAVDNDTGMNAEVRYSIVGGNT72 V.2 690INVVDVNDNKPVFIVPPSNCSYELVLPSTNPGTVVFQVIAVDNDTGMNAEVRYSIVGGNT74 V.1 721 RNDVNADNETGNIMEKCSV'rLGVLTVVNDLGQPDSLFVINLFVRESIVNT 700 V.1 721RDLFAIDQETGNITMEKCDVTDLGLHRVLVKANDLGQPDSLFSVVIVNLFVNESVTNAT78 V.2 750RDLFAIDQETGNITLMEKCDVTDLGLHRVLVKANDLGQPDSLFSVVIVNLFVNESVTNAT80 V.21 781 LINLVRKSQTEAPVTPNEIDVSTSDYHRVKINLVAAVAGTITVVVIFAVVRAP 809 V.1 781LINELVRKSTEAPVTPNTEIADVSSFTSDYVKILVAAVAGTITVVVVI FITAVVRCROAP84 V.2 810LINELVRKSTEAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVVVIFITAVVRCRQAP86 00 00 V.1 841 HLKAAQKNKQNSEWATPNPENRQMIMMI4KKXKIHSPKNLLLNFVTIEETKADDVDSDG 900

HLKAAQKNKQNSEWATPNPENRQMIMMKKKKKKKHSPKNLLLNFVTIEETKADDVDSDG

V.2 870 HLKAAQKNKQNSEWATPNPENRQMIMMKKIKKKKHSPKNLLLNFVTIEETKADDVDSDG 929 V.1 901 NRVTLDLPIDI.EEQTMGKYNWVTTPTTFKPDSPDLARHiYKSASPQPAFOIQPETPLNSKH 960

NRVTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLARHYKSASPQPAFQIQPETPLNSKH

V.2 930 NRVTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDfLAPHYKSASPQPAFQIQPETPLNSKH 989 V.1 961 HIIOELPLDNTFVACDSISKCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRPVGIQVS 1017 HIIQELPLDNTFVACDSISKCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP S V.2 990 HIIQELPLDITEVACDSISKCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRPTDSRTS 1046 Table 1-11(b). Nucleotide sequence of transcript variant 1 09PI D4 v.3 (SEQ ID NO: 243) ctggtggtcc agtacctcca aagatatgga atacactcct gaaatatcct g ttttttcaga atcctttaat aagcagttat gtcaatctga aagttgctta c atattaatag ctattcttgt ttttcttatc caaagaaaaa tcctctaatc c atgatagttg ttaccatgtt taggcattag tcacatcaac ccctctcctc t ctcttcttca aatcaaactt tattagtccc tcctttataa tgattccttg c tccagatcaa ttttttttca ctttgatgcc cagagctgaa gaaatggact a ttattcattg ccaagagaat aattgcattt taaacccata ttataacaaa g tatattttgt gatttgtaac aaataccctt tattttccct taactattga a ttaattattt gtattctctt taactatctt ggtatattaa agtattatct t tatcaatggt ggacactttt ataggtactc tgtgtcattt ttgatactgt a tttcatttat ctttattctt aatgtacgaa ttcataatat ttgattcaga a cactaattaa cagagtgtca attatgctaa catctcattt actgatttta a gtttttgtta acatgcatgt ttagggttgg cttcttaata atttcttctt c ctctcctctt cttttggtca gtgttgtgcg ggttaataca acaaactgta a ctggtatgga cttgttgtcc gggacgtaca ttttcgcggt cctgctagca t tccactctgg cgcccaggag aaaaactaca ccatccgaga agaaatgcca g tgataggcga cttgttgaaa gaccttaact tgtcgctgat tccaaacaag t ctgctatgca gttcaagcta gtgtacaaga ccggagatgt gccactgatt c aggatactgg tgagatcttc actactggcg ctcgcattga tcgtgagaaa t gtatcccaag ggatgagcat tgcttttatg aagtggaggt tgccattttg c tatttagact ggttaagata cgttttctga tagaagatat aaatgataat g tcccagcaac agttatcaac atatcaattc cagagaactc ggctataaac t ctctcccagc ggctgttgat cctgacgtag gaataaacgg agttcaaaac t ttaagagtca aaacattttt ggcctcgatg tcattqaaac accagaagga g cacaactgat tgttcaaaag gagttagata gggaagagaa ggatacctac g taaaggttga agatggtggc tttcctcaaa gatccagtac tgctattttg c ttactgatac aaatgacaac cacccagtct ttaaggagac agagattgaa g cagaaaatgc tcctgtaggc acttcagtga cacagctcca tgccacagat g gtgaaaatgc caagatccac ttctctttca gcaatctagt ctccaacatt g tatttcacct caatgccacc actggactta tcacaatcaa agaaccactq g aaacaccaaa ccacaagtta ctggttttgg caagtgatgg tggattgatg c caatggtgct ggtaaatgtt acagatgtca atgataatgt cccatccatt g acatcgtcaa tcctgtcaat gacacagttg ttctttcaga aaatattcca c aaattgctct cataactgtg acggataagg atgcggacca taatggcagg g tcacagatca tgaaatccct ttcagattaa ggccagtatt cagtaatcag t agactgcagc atatcttgac tatgagtcca caaaagaata tgccattaaa t cagatgctgg caaacctcct ttgaatcagt cagcaatgct cttcatcaaa g aaaatgacaa tgctccagtt ttcacccagt ctttcgtaac tgtttctatt c actctcctgg catccagttg acgaaagtaa gtgcaatgga tgcagacagt g ctaagatcaa ttacctgcta ggccctgatg ctccacctga attcagcctg g caggcatgat gactgtagtg aagaaactag atagagaaaa agaggataaa t caattctggc aaaagataac ggggtaccac ccttaaccag caatgtcaca g gcattattga. tcagaatgac aatagcccag ttttcactca caatgaatac a tcccagaaaa ccttccaagg catggtacag taggactaat cactgtaact g atggagacaa ttctgcagtt acgctctcca ttttagatga gaatgatgac t attcacaaac tggtgtcatc cgaccaaata. tttcatttga tagagaaaaa c acactttcta tgtaaaggct gaggatggtg gtagagtatc acgttcttca a taaccataaa tgtggttgat gtcaatgaca acaaaccagt tttcattgtc c raaaactttt :ttgtacttt :ccttttcac .cccaaactt ctcgtttta .ctgtataaa raataatgat .ttaaatatt ttatatatt ggtatctta caaatttat tttaaaaca :ctcttctct caagtgtac gcgtggtgt aaaacgtcc ccttgacaa :gaattgaag tatgtgctg cggatgaaa caccattgt ctaaatata acgaactaa acaagatgc tgatgaaag aagtgagtg tcagtatac ctgacatag ccaggagat atagggaag cagcaagag acataagat tcaacacca tgacatgct tcctcctgg tactggctg tgaaagatg ctgagaata ggcctaatg attgtcgta atttattca tctttgtaa acttctatg atcctgatt tcaccattg aagaatctt gtgccaaag ctccttcca 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 00 00 actgttctta ttgctgttga acacaagaga gtgatgttac ctgattctct ctacactgat agatagctga ctggcaccat caccacacct cagaaaacag agaacttgct atggaaacag acaattgggt acaaatctgc agcaccacat ccaagtgttc cgaccttcga cttgcacccc ggaaatctga aaggctctca ccagcacatc cacccagcaa gactaaagaa gcactcaaga atcattccag aggcctctac ctccagtgac tccaccacag aggcctcagc ctcctctgcc agcaaggttg gtgcaacatc aagtcattcc cccccattat agatgtatat taaataacta atagttaacc atttaacaat gatgtacagt gctgattgtg gtttaccttt gccttttttt tagtgagtct ctgctgtcat ttttcaataa ctttagtaga aaaaagggcc tctgccattt gtaacagcct tcgacggtaa ctcaataggc aaaaccatat aagaaaacca aaatctacat gcttcttgca catatatgaa tagttttctt tatgtgcaca actgagaaaa gggcggttgg tgtaatgtat actgtggtaa tgaattggtt caatgacact tctgtttgca agaccttggt cttcagtgtt taatgaactg tgtatcctca aactgtcgtt taaggctgct gcagatgata gcttaatttt agtcacacta aactacacct ctctccacag catccaagaa ctcaagcagt ggtacctgtg catgaaagag agggaaagtg ggaaagcagc tcatggcctg tcgcactgaa agctgcagaa atgtctcatc ctcttcqcaa tcagcaccac acagaccatc tcctcctcta cctctgctac acaggttatt ggtgcaaggt tcagttttac tttgacaacc ggaagaacat agtcaaaatt tgtaaataga aaaaaattgc ttgcatcccc attttttgtt tggaaccagt agtttaccta gggctttctt cccttcaaaa gttttctacc agaatatgta gttcgagaca tctttttact taagcctgta agaaaattaa actactattt tgttaccaaa actaaatcta ctaataacaa caaacagaat gtcaacaaga ataaaatata ctgttgtctg ttttataaat taccttgttt ggtggggggg aacttgatat ttgcataatt ctaccgtcca ggcatgaatg atcgaccaag ttacacagag gtaattgtca gtgcgcaaaa ccaactagtg gtagttattt cagaaaaaca atgatgaaga gtcactattg gaccttccta actactttca cctgccttcc ctgcctctcg tcagatccct tccgtacaca tctacaacta gcaggaaagt agtgatggtg ccccttggct ggggatggca ataactgttc tatggccatt gcacaggcct agcccacgag gcattgtgcc gtgcaggcta agccctcctt gccctccatc gctgatgggc accatgtctg ttcactccac cccttgtaaa taagatacaa aacagatacc aatttgttta ttgtacagta gtttttatca atgtagcaaa aacttttgtt tttgattttt tacgcagtag ttattccaat taaactgtac cagaagtgca ttaatagttt gtccattatt aaggaggtgg tgtagagaaa gggtgttcag taagactaag caagaatatc tttaaggaaa aataccaata ttcttctaag ttatttcctt gtacagaaac cagtataaca ggagtcaata ttaatttatg tcattggtga ctaatccagg cagaggttcg aaacaggcaa tgttggtcaa atctgttcgt gcactgaagc actatgtcaa tcatcactgc agcagaattc aaaagaaaaa aagaaactaa ttgatctaga agcccgacag aaattcagcc ataacacctt acagcgtttc ccagaccgcc tggagatctg cccagcggcg gactgggaga atcctcagga actccgatcc aaccaactgt ctgatgcctq ctgctctatg tgacacagac acagcccacc ctgcacttca tagcacaggc gtagtcaggc tatgctctgt aaagacttca gccaacaqgc gctaaaatag ttccaatgag agaataaatc attcagaatg aggcttatca tcatgtgcaa tggaaagcct cagataacgt gtttgttgtt gtagtgtaaa actatattgt agatctagat ataactgccc agtgtaaagt acttgggtct atgcatccaa ctcaggaaga taaaaataac ggatttttgt aggaaggaac aatgcagagg acacacacag caaagaaaca cttgtatcct atcaccaact ctaaaccaag tctcctattg attaaactgt ggatttccac cacagtqgtc ttacagcatt cataacattg agctaatgac gaatgagtcg accagtgacc gatcctggtt tgtagtaaga tgaatgggct gaagaagaag ggcagatgat agagcaaaca ccctgatttg tgaaactccc tgtggcctgt tgactgtggc aatgaaggag gattcatccc tgtcacattt ccatgatgca ggagtacttt tgaatctact ggaagaggcc ctggatgccg ccacagccca cattgctctc accgatacag ccacagccca tgctgcaatc ccaatcatca tgatcaggga tcccagtgat cagaccgtcc ttacttcaaa tattctgatt tacagctaga tgtatttaaa tgacagagcg tattactgat agaaatatct taaaaggtat ttcagttttt tactgcttgt tgataaaatt ctacaaccta taattaagca acatcagaaa ttacttctgg agcacgagtc tttaaatgtt aaatacatgt tattctagct ttttcaagaa gagaaataag aacaaaaacc gtactattca cttaactggc taattttctt agacaattga attaacttag gtgtaaattt tgaatattga tttcaggtaa gtaggaggaa atggagaaat ttaggacagc gtgaccaatg ccaaatactg gcagctgttg tgtcgccagg accccaaacc cattccccta gttgacagtg atgggaaagt gcccgacact ctgaattcga gactctatct tatccagtga gttgtgcgat caaccacagc cacctgccag ggcagcctta gatcgtgcta ttcatacctg tctgacaact gcatctctgg ccactqtcac tgccacagcc gtgtctgct c ccatcagcac agccacagct gtcagtttgc gtgcaaggta gattcaatta agaggtgatt ttttcagaaa atcagatttg cccttagtca aagaaaagga cactatttct ttgtttccat tattttctaa acgtactcta ttgttgttgt ttgtgtctct tgtatataca tttctctact actatttgtt taaagctgta gaatttgtat acttaaaata gatttgacag aactgtagat caacttactg atgtaattat gcacatgact atcaaaatct tagaaaacat cattatcttg ccatagcaaa tgtttaatgg acatagattt tgtaacataa gaaagtttct 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 5940 6000 6060 6120 6180 6240 6300 6360 6420 6480 6540 6600 7-- 00 00 tttcatgtgc aaagttcctt agagtttgat ggatttggag aatgtttatt aaattttaaa agtaqatttt ttgtgtttta acaCacaCac gttgaaagat aatcaaatag agttactagg cctatagatg ataatcagac taccagagag cccagttatt gaaacctgtt atttctctga tgtatgatgt ggttctgttt atatgagaat attgtgatta aaatactaag ttttaactaa taataaattt tgtaaatttg acaaagagcc aaatagacta aaaaaatcct attagttttt tacacattta tttattataa aqttagttta ggctcaattg ctttctatat ttctttcaca aaagcagttc atgtgcacaa aagagtaaac tatgtattta aagtgaacag gtatggccaa aggttctgta ccagcaggtt aagtcatatt taataaaatt catactggtt aaattattat cttggggaag agaacattct attattttca acacacacac tattaaataa gactcaaagt gaggttctta atataggaat ttagattgtg gctttgaatg ctcccctatg acttctaggg ctgaatagac atgtagtcct taaaaaggat aatacgttca ttaattttct tatcttatat tattgtgtca agagagtaat gttagaaaat tctgccttcc aacaaatcac tttcaatcct tttgtttgtt tgaaaatcta ttattatttc ggtaaacttt gccgggaaa a tatgtgccaa gtagctttct aagttgacaa ataaaaaaaa tgtgtgaaat tgtttgtatt tatcccaaag ggatggcagt ttgcatgttt aagtagcqtt tqactgggcg aattaatgtt ttccaggtgc catacaaatg attaagaaaa ttgcctcaaa cattttctct gtgcacacac cttatcaggc tggatatttg aatcctcata gaaccaatac tttagaatat gaagcaggct ccttccttct cttcagatct agtggtatag acaactgcaa tttgtttgat atcaaagtag agctatggta gctacgtgca gtgtattaaa tttgtgtatt aataataaat caaactaata aaattgttca gagaaaatta tctttttcat cttgtttagg ttatctttct ttattatgac catgggagca ttaccacacc tacactaacc cagcagaaac ttaatggcaa ttatactatc gtgggaagat cagttccaac atgtaatcca tcttattaat ttcagaatat tgcagaataa ttttctcctt atgtgaatcc tgttgatatt gaaccaatag aaacctgcaa ctgcaaaact acacacattt atctcaatgg ggatttttct tctggaaact gcttttatta taaatgactg gagagtagcc ctttctaagc gatgatatct gttgacacag aacgtcttac ttgaaattaa ttattctatt ttactatatc tacacattct aattagcttt cttatttact ggttagtgct tttatcacac gttcttaaaa aaggcgtttt ggtattactg cttttattta tcttttatat cattagaaac agagaagctg agatcaattt gtcatgtgct agtaacaaaa ttgtttagtg cctgcttaaa tcctcctctg catgctttgg gaagcaaact atatattaat acattattcc cttcttaact cgtgttgtta cgaaggactg gtggctattg tgacaaaaat agatgatgtg ttagttttct aaatgatata ttactatcta tctgacagta tgtgacgttt ccctttctaa ggcaccctct aaagaggcaa gtccactagg ttttcatcac cacacaagtg tgaaccaaca aacttcaagc ttgtgtccat acacttgtga tttcttaaac tacatatgat taacatttta attgtgtaat atggtcatta tgtaattatg actcacatgg aaggtgtgta tactcttctg tttttggaaa tattttgaat aaatatattt tatgcagagg tttagtaaat aaatctgctc attgtaagtg atattaagat tgatatcata aagtaagaag tgtat taatt aaaagttatg catccattgt tttaactatc atgttccaag atgatatttg ttgatqttga cagtgcttcc agattttttc gatgatctac aaaagaagag tgttagtgaa taatttattg tgacacc~t ctctgatttt tcttggtttt ggggtattag tctggccttg attacaagtt gctattgtga atcaaaaaat tgaatgactt attccattag gtatgtattc tttacctgtg atctacaatg cttttaatta ggtagcagtt aatgggaaaa tcacacacac ctatttcaac tactccctaa atttatattt eca aatt tat gcttccaact ctgcaagaac ccttaaaata atgattttta agaaaaatgt atacttttta ttttatgaaa cagcatctga gttgactatt gttctatttc agaaat 6660 6720 6780 6840 6900 6960 7020 7080 7140 7200 7260 7320 7380 7440 7500 7560 7620 7680 7740 7800 7860 7920 7980 8040 8100 8160 8220 8280 8340 8400 8460 8520 8580 8640 8700 8760 8820 8880 8940 9000 9060 9120 9176 Table 1.11(b). Nucleotide sequence alignment of 109)1D4 v.1 (SEQ ID NO: 244) and 109NID4 v.3 (SEQ ID NO: 245) Score =7456 bIts (3878), Expet =0.01dentfies 387813878 (100%) Strand Plus I Plus V.1 1 ctggtggtccagtacctccaaagatatggaatacactcctgaaatatcctgaaaactttt V.3 1 ctggtggtccagtacctccaaagatatggaatacactcctgaaatatcctgaaaactttt V.1 61 ttttttcagaatcctttaataagcagttatgtcaatctgaaagttgcttacttgtacttt 120 V.3 61 ttttttcagaatcctttaataagcagttatgtcaatctgaaagttgcttacttgtacttt 120 V.1 121 atattaatagctattcttgtttttcttatccaaagaaaaatcctctaatccccttttcac 180 V.3 121 atattaatagctattcttgtttttcttatccaaagaaaaatcctctaatccccttttcac 180 V.1 181 atgatagttgttaccatgtttaggcattagtcacatcaacccctctcctctcccaaactt 240 251 00V.3 181 atgatagttgttaccatgtttaggcattagtcacatcaacccctctcctctcccaaactt 240 V.1 241 ctcttcttcaaatcaaactttattagtccctcctttataatgattccttgcctcgtttta 300 V.3 241 ctcttcttcaaatcaaactttattagtccctcctttataatgattccttgcctcgtttta 300 tnV.1 301 tccagatcaattttttttcactttgatgcccagagctgaagaaatggactactgtataaa 360 V.3 301 tccagatcaattttttttcactttgatgcccagagctgaagaaatggactactgtataaa 360 V.1 361 ttattcattgccaagagaataattgcattttaaacccatattataacaaagaataatgat 420 V.3 361 ttattcattgccaagagaataattgcattttaaacccatattataacaaagaataatgat 420 C1V.1 421 tatattttgtgatttgtaacaaataccctttattttcccttaactattgaattaaatatt 480 00 11111111111111111111! ii111111 V.3 421 tatattttgtgatttgtaacaaataccctttattttcccttaactattgaattaaatatt 480 V.1 481 ttaattatttgtattctctttaactatcttggtatattaaagtattatcttttatatatt 540 V.3 481 ttaattatttgtattctctttaactatcttggtatattaaagtattatcttttatatatt 540 V.1 541 tatcaatggtggacattttataggtactctgtgtcatttttgatactgtaggtatctta 600 V.3 541 tatcaatggtggacacttttataggtactctgtgtcatttttgatactgtaggtatctta 600 V.1 601 tttcatttatctttattcttaattacgaattcataatatttgattcagaacaaatttat 660 V.3 601 tttcatttatctttattcttaatgtacgaattcataatatttgattcagaacaaatttat 660 V.1 661 cactaattaacagagtgtcaattatgctaacatctcatttactgattttaatttaaaaca 720 V.3 661 cactaattaacagagtgtcaattatgctaacatctcatttactgattttaatttaaaaca 720 V.1 721 gtttttgttaacatgcatgtttagggttggcttcttaataatttcttcttcctcttctct 780 V.3 :721 gtttttgttaacatgcatgtttagggttggcttcttaataatttcttcttcctcttctct 780 V.1: 781 ctctcctcttcttttggtcagtgttgtgcgggttaatacaacaaactgtaacaagtgtac 840 V.3: 781 ctctcctcttcttttggtcagtgttgtgcgggttaatacaacaaactgtaacaagtgtac 840 V.1 :841 ctggtatggacttgttgtccgggacgtacattttcgcggtcctgctagcatgcgtggtgt 900 V.3 ;841 ctggtatggacttgttgtccgggacgtacattttcgcggtcctgctagcatgcgtggtgt 900 V.1 :901 tccactctggcgcccaggagaaaaactacaccatccgagaagaaatgccagaaaacgtcc 960 V.3 :901 tccactctggcgcccaggagaaaaactacaccatccgagaagaaatgccagaaaacgtcc 960 V.1 :961 tgataggcgacttgttgaaagaccttaacttgtcgctgattccaaacaagtccttgacaa 1020 V.3 :961 tgataggcgacttgttgaaagaccttaacttgtcgctgattccaaacaagtccttgacaa 1020 V.1 :1021 ctgctatgcagttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaag 1080 252 V.3 1021 ctgctatgcagttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaag 1080 V.1 1081 aggatactggtgagatcttcactactggcgctcgcattgatcgtgagaaattatgtgctg 1140 V.3 1081 aggatactggtgagatcttcactactggcgctcgcattgatcgtgagaaattatgtgctg 1140 V.1: 1141 gtatcccaagggatgagcattgcttttatgaagtggaggttgccattttgccggatgaaa 1200 V.3: 1141 gtatcccaagggatgagcattgcttttatgaagtggaggttgccattttgccggatgaaa 1200 V.1 ;1201 tatttagactggttaagatacgttttctgatagaagatataaatgataatgcaccattgt 1260 riV.3 :1201 tatttagactggttaagatacgttttctgatagaagatataaatgataatgcaccattgt 1260 00 V.1 :1261 tcccagcaacagttatcaacatatcaattccagagaactcggctataaactctaaatata 1320 V.3 :1261 tcccagcaacagttatcaacatatcaattccagagaactcggctataaactctaaatata 1320 V. 31ccccgcgttgtcgctagaaagatcaacagata18 V.1 1321 ctctcccagcggctgttgatcctgacgtaggaataaacggagttcaaaactacgaactaa 1380 V.1: 1381 ttaagagtcaaaacatttttggcctcgatgtcattgaaacaccagaaggagacaagatgc 1440 V.3: 1381 ttaagagtcaaaacatttttggcctcgatgtcattgaaacaccagaaggagacaagatgc 1440 V.1 :1441 cacaactgattgttcaaaaggagttagatagggaagagaaggatacctacgtgatgaaag 1500 V.3 :1441 cacaactgattgttcaaaaggagttagatagggaagagaaggatacctacgtgatgaaag 1500 V.1 :1501 taaaggttgaagatggtggctttcctcaaagatccagtactgctattttgcaagtgagtg 1560 V.3 1501 taaaggttgaagatggtggctttcctcaaagatccagtactgctattttgcaagtgagtg 1560 V.1 1561 ttactgatacaaatgacaaccacccagtctttaaggagacagagattgaagtcagtatac 1620 V.3 1561 ttactgatacaaatgacaaccacccagtctttaaggagacagagattgaagtcagtatac 1620 V.1 1621 cagaaaatgctcctgtaggcacttcagtgacacagctccatgccacagatgctgacatag 1680 V.3 1621 cagaaaatgctcctgtaggcacttcagtgacacagctccatgccacagatgctgacatag 1680 V.1 1681 gtgaaaatgccaagatccacttctctttcagcaatctagtctccaacattgccaggagat 1740 V.3 1681 gtgaaaatgccaagatccacttctctttcagcaatctagtctccaacattgcaggagat 1740 V.1: 1741 tatttcacctcaatgccaccactggacttatcacaatcaaagaaccactggatagggaag 1800 V.3: 1741 tatttcacctcaatgccaccactggacttatcacaatcaaagaaccactggatagggaag 1800 V.1 :1801 aaacaccaaaccacaagttactggttttggcaagtgatggtggattgatgccagcaagag 1860 V.3: 1801 aaacaccaaaccacaagttactggttttggcaagtgatggtggattgatgccagcaagag 1860 253 00V.1 16 atggtgaagtcgttataattcacataaagt12 V.3 1861 caatggtgctggtaaatgttacagatgtcaatgataatgtcccatccattgacataagat 1920 1921 acatcgtcaatcctgtcaatgacacagttgttctttcagaaaatattcactcaacacca 1980 V.3 :1921 acatcgtcaatcctgtcaatgacacagttgttctttcagaaaatattccactcaacacca 1980 V.1: 1981 aaattgctctcataactgtgacggataaggatgcggaccataatggcagggtgacatgct 2040 V.3: 1981 aaattgctctcataactgtgacggataaggatgcggaccataatggcagggtgacatgct 2040 V. 01taaacagatcttaatagcattcgatcgtcctg20 V.31 2041 tcacagatcatgaaatccctttcagattaaggccagtattcagtaatcagttcctcctgg 2100 00 V.1 :2101 agactgcagcatatcttgactatgagtccacaaaagaatatgccattaaattactggctg 2160 (iV.3 :2101 agactgcagcatatcttgactatgagtccacaaaagaatatgccattaaattactggctg 2160 V.1 :2161 cagatgctggcaaacctcctttgaatcagtcagcaatgctcttcatcaaagtgaaagatg 2220 V.3 :2161 cagatgctggcaaacctcctttgaatcagtcagcaatgctcttcatcaaagtgaaagatg 2220 V.1: 2221 aaaatgacaatgctccagttttcacccagtctttcgtaactgtttctattcctgagaata 2280 V.3: 2221 aaaatgacaatgctccagttttcacccagtctttcgtaactgtttctattcctgagaata 2280 V.1 :2281 actctcctggcatccagttgacgaaagtaagtgcaatggatgcagacagtgggcctaatg 2340 V.3 2281 actctcctggcatccagttgacgaaagtaagtgcaatggatgcagacagtgggcctaatg 2340 V.1 2341 ctaagatcaattacctgctaggccctgatgctccacctgaattcagcctggattgtcgta 2400 V.3 2341 ctaagatcaattacctgctaggccctgatgctccacctgaattcagcctggattgtcgta 2400 V.1 2401 caggcatgctgactgtagtgaagaaactagatagagaaaaagaggataaatatttattca 2460 V.3 2401 caggcatgctgactgtagtgaagaaactagatagagaaaaagaggataaatatttattca 2460 V.1 :2461 caattctggcaaaagataacggggtaccacccttaaccagcaatgtcacagtctttgtaa 2520 V.3 :2461 caattctggcaaaagataacggggtaccacccttaaccagcaatgtcacagtctttgtaa 2520 V.1 :2521 gcattattgatcagaatgacaatagcccagttttcactcacaatgaatacaacttctatg 2580 V. 3 :2521 gcattattgatcagaatgacaatagcccagttttcactcacaatgaatacaacttctatg 2580 V.1 :2581 tcccagaaaaccttccaaggcatggtacagtaggactaatcactgtaactgatcctgatt 2640 V.3 :2581 tcccagaaaaccttccaaggcatggtacagtaggactaatcactgtaactgatcctgatt 2640 V.1 :2641 atggagacaattctgcagttacgctctccattttagatgagaatgatgacttcaccattg 2700 V.3 :2641 atggagacaattctgcagttacgctctccattttagatgagaatgatgacetcaccattg 2700 254 00 V.1 2701 attcacaaactggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatctt 2760 riV.3 2701 attcacaaactggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatctt 2760 V.1 2761 acactttctatgtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaag 2820 V.3 2761 acactttctatgtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaag 2820 V.1 2821 taaccataaatgtggttgatgtcaatgacaacaaaccagttttcattgtccctccttcca 2880 V.3 2821 taaccataaatgtggttgatgtcaatgacaacaaaccagttttcattgtccctccttcca 2880 riV.1 2881 actgttcttatgaattggttctaccgtccactaatccaggcacagtggtctttcaggtaa 2940 riV.3 2881 actgttcttatgaattggttctaccgtccactaatccaggcacagtggtctttcaggtaa 2940 00 V.1 2941 ttgctgttgacaatgacactggcatgaatgcagaggttcgttacagcattgtaggaggaa 3000 V.3 2941 ttgctgttgacaatgacactggcatgaatgcagaggttcgttacagcattgtaggaggaa 3000 V.1 3001 acacaagagatctgtttgcaatcgaccaagaaacaggcaacataacattgatggagaaat 3060 V.3 3001 acacaagagatctgtttgcaatcgaccaagaaacaggcaacataacattgatggagaaat 3060 V.1 3061 gtgatgttacagaccttggtttacacagagtgttggtcaaagctaatgacttaggaagc 3120 V.3 3061 gtgatgttacagaccttggtttacacagagtgttggtcaaagctaatgacttaggacagc 3120 V.1 3121 ctgattctctcttcagtgttgtaattgtcaatctgttcgtgaatgagtcggtgaccaatg 3180 V.3 3121 ctgattctctcttcagtgttgtaattgtcaatctgttcgtgaatgagtcggtgaccaatg 3180 V. 1 3181 ctacactgattaatgaactggtgcgcaaaagcactgaagcaccagtgaccccaaatactg 3240 V.3 3181 ctacactgattaatgaactggtgcgcaaaagcactgaagcaccagtgaccccaaatactg 3240 V.1 3241 agatagctgatgtatcctcaccaactagtgactatgtcaagatcctggttgcagctgttg 3300 V.3 3241 agatagctgatgtatcctcaccaactagtgactatgtcaagatcctggttgcagctgttg 3300 V.1 3301 ctggcaccataactgtcgttgtagttattttcatcactctgtagtaagatgtcgccagg 3360 V.3: 3301 ctggcaccataactgtcgttgtagttattttcatcactgctgtagtaagatgtcgccagg 3360 V.1 :3361 caccacaccttaaggctgctcagaaaaacaagcagaattctgaatgggctaccccaaacc 3420 V. 3 :3361 caccacaccttaaggctgctcagaaaaacaagcagaattctgaatgggctaccccaaacc 3420 V.1 :3421 cagaaaacaggcagatgataatgatgaagaaaaagaaaaagaagaagaagcattccccta 3480 V.3 :3421 cagaaaacaggcagatgataatgatgaagaaaaagaaaaagaagaagaagcattccccta 3480 V.1: 3481 agaacttgctgcttaattttgtcactattgaagaaactaaggcagatgatgttgacagtg 3540 V.3: 3481 agaacttgctgcttaattttgtactattgaagaaactaaggcagatgatgttgacagtg 3540 00 V.1: 3541 atggaaacagagtcacactagaccttcctattgatctagaagagcaaacaatgggaaagt 3600 V.3: 3541 atggaaacagagtcacactagaccttcctattgatctagaagagcaaacaatgggaaagt 3600 V.1: 3601 acaattgggtaactacacctactactttcaagcccgacagccctgatttggcccgacact 3660 V.3: 3601 acaattgggtaactacacctactactttcaagcccgacagccctgatttggcccgacact 3660 V.1 :3661 acaaatctgcctctccacagcctgccttccaaattcagcctgaaactcccctgaattcga 3720 V.3 :3661 acaaatctgcctctccacagcctgccttccaaattcagcctgaaactcccctgaattcga 3720 V.1 :3721 agcaccacatcatccaagaactgcctctcgataacacctttgtggcctgtgactctatct 3780 V.3 :3721 agcaccacatcatccaagaactgcctctcgataacacctttgtggcctgtgactctatct 3780 V.1 :3781 ccaagtgttcctcaagcagttcagatccctacagcgtttctgactgtggctatccagtga 3840 V.3: 3781 ccaagtgttcctcaagcagttcagatccctacagcgtttctgactgtggctaccagtga 3840 V.1 3841 cgaccttcgaggtacctgtgtccgtacacaccagaccg 3878 V.3 3841 cgaccttcgaggtacctgtgtccgtacacaccagaccg 3878 Table LIV(b). Peptide sequences of protein coded by 109P1 D4 v.3 (SEQ ID NO: 246) MDLLSGTYIF AVLLACVVFII SGAQEKNYTI REEMPENVLI GDLLKDLNLS MQFKLVYKTG DVPLIRIEED TGEIFTTGAR IDREKLCAGI PRDEHCFYEV RLVKIRFLIE DINDNAPLjFP ATVINISIPE NSAINSKYTL PAAVDPDVGI SQNI FGLDVI ETPEGDKMPQ LIVQKELDRE EKDTYVMKVK VEDGGFPQRS DTNDNHPVFK ETEIEVSIPE NAPVGTSVTQ LHATDADIGE NAKIHFSFSN HLNATTGLIT IKEPLDREET PNHKLLVLAS DGGLMPARAN VLVNVTDVND VNPVNDTWVL SENIPLNTKI ALITVTDKDA DHNGRVTCFT DHEIPFRLRP AAYLDYESTK EYAIKLLAAD AGKPPLNQSA MLFIKVKDEN DNAPVFTQSF PGIQLTKVSA MDADSGPNAK INYLLGPDAP PEFSLDCRTG MLTVVKKLDR LAKDNGVPPL TSNVTVFVSI IDQNDNSPVF THNEYNFYVP ENLPRHGTVG DNSAVTLSIL DENDDFTIDS QTGVIRPNIS FDREKQESYT FYVKAEDGGR INVVDVNDNK PVFIVPPSNC SYELVLPSTN PGTVVFQVIA VDNDTGMNAE RDLFAIDQET GNITLMEKCD VTDLGLHRVL VKANDLGQPD SLFSVVIVNL LINELVRKST EAPVTPNTEI ADVSSPTSDY VKILVAAVAG TITVVVVIFI HLKAAQKNKQ NSEWATPNPE NRQMIMMKKK KKKKKHSPKN LLLNFVTIEE NRVTLDLPID LEEQTMGKYN WVTTPTTFKP DSPDLARHYK SASPQPAFOI HIIQELPLDN TFVACDSISK CSSSSSDPYS VSDCGYPVrT FEVPVSVHTR TPMKESTTME IWIHPQPQRK SEGKVAGKSQ RRVTFHLPEG SQESSSDGGL TSHGLPLGYP QEEYFDRATP SNRTEGDGNS DPESTFIPGL KKAAEITVQP QECLIYGHSD ACWMPASLDH SSSSQAQASA LCHSPPLSQA STQHHSPRVT VTQTIALCHS PPPIQVSALH HSPPLVQATA LHHSPPSAQA SALCYSPPLA LPQVIALHRS OAQSSVSLQQ GWVQGADGLC SVDQGVQGSA TSQFYTMSER IPLTTFTPRQ QARPSRGDSP IMEEHPL LI PNKSLTTA

EVAILPDEIF

NGVQNYELIK

STAILQVSVT

LVSNIARRLF

NVPSIDIRYI

VFSNQFLLET

VTVSIPENNS

EKE OKYL FT I

LITVTDPDYG

VSRSSSAKVT

VRYSIVGGNT

FVNESVTNAT

TAVVRCRQAP

TKADDVDS OG

OPETPLNSKH

PPMKEVVRSC

GDHDAGSLTS

TVEEASDNCT

QTIALCHSPP

QAAAISHSSP

LHPSDDSIKV

120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1347 Table LV(b). Amino acid sequence alignment of 109PI D4 v.1 (SEQ ID NO: 247) and 109PI D4 v.3 (SEQ 10 NO: 248) Score 2005 bits (5195), Expect =0.Oldentilies =101111011 Positives =101111011 (100%) V.1 1 MDLLSGTYIFAVLLACVVFHSGAQEKNYTIREEMPENVLIGDLLKDLNLSLIPNKSLTTA

MDLLSGTYIFAVLLACVVFHSGAQEKNYTIREEMPENVLIGDLLDLNLSLIPNKSLTTA

V.3 1 MDLLSGTYIFAVLLACVVFHSGAQEKNYTIREEMPENVLIGDLLKDLNLSLIPNKSLTTA V.1 61 MQFKQ.VYKTGDVPLIRIEEDTGEIrTTGARIDREKLCAGIPRDEHCFYEVEVAILPDEIF 120 oV.3 :61 MQEKLVYKTGDVPLIRIEEDTGEIFTTGARIDREKLCAGIPRDEHCFYEVEVAILPDEIF 120 V.1 :121 RLVKIRFLIEDINDNAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVQNYELIK 180

RLVKIRFLIEDINDNAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVQNYELIK

V.3: 121 RLV1IRFLIEDINDNAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVQNYELIK 180 V. 8 CIGDITEDMoIQEDREDYMVVDGPRSALVV 4 V.1 181SQNIFGLDVIETPEGDKNPQLIVQKELDREEKDTYVMKVKVEDGGFPQRSSTAILQVSVT24 V.3 181SQNIFGLDVIETPEGDKMPQLIVQKELDREEKDTYVMKVCVEDGGFPQRSSTAILOVSVT24 OV.1 1821 SONIFGLVIETPIIEGD PQLVKEVLDREEKDIMKVKDFQSSTLOVSVTL 2400 V.1 241DTNDNHPVFKETEIEVSIPENAPVGTSVTQLHATDADIGENAKIHFSFSNLVSNIARRLF30 ciV.3 :241 DTNDNHPVFKETEIEVSIPENAPVGTSVTQLHATDADIGENAKIHFSFSNLVSNIARRLF 300 0V.1 :301 HLNATTGLITIKEPLDREETPNXLLVLASDGGLMPARAMVLVNVTDVNDNVPSIDIRYI 360 V.3 :301 HLNATTGLITIKEPLDREETPNHCLLVLASDGGLMPARAMVLVNVTDVNDNVPSIDIRYI 360 00V.1 :361 VNPVNDTVVLSENIPLNTKIALITVTDKDADHNGRVTCFrDHEIPFRLRPVFSNQELLET 420

ONVDVLEILTILTTKADNRTFDEPRRVSQLE

OV.3 :361 VNPVNDTVVLSENIPLNTKIAILITVTDKDADHNGRVTCFIDHEIPFRLRPVFSNOFLLET 420 V. 2 ALYSKYIcADGPiQSMFKKEDAVrSVVIEN 8 V.1 421AAYLDYESTKEYAIK(LLAADAGKPPLNQSAHLFIKVKDENDNAPVFTQSFVTVSIPENNS48 V.3 421AAYLDYESTKEYAIKLLAADAGKPPLNQSAMLFIKVKDENDNAPVFIQSFVTVSIPENNS48 V.3 421 AAYQLDTKYAIKLLAADNAGKPLLPNQAMPFLKED NAPMLVFQSFVSENNSE 540 V.1 481PGIQLTPCVSANDADSGPNAKINYLLGPDAPPEFSLDCRTGMLTVVKKLDREKEDKYLFII54 V.3 481PGIQLTKVSAMDADSGPNAKINYLLGPDAPPEFSLDCRTGtdLTVVKKLDREKEDKYLFTI54 V.1 541 PLKVSMDADSNIDNLDPFHEPESRGLVHVKLEEDY 500 V.1 541 V.3 541LAKDNGVPPLTSNVTVFVSIIDQNDNSPVrTHNEYNFYVPENLPRHGTVGLITVTDPDYG60 V.3 541 LDNVLTSNVTVFVIIDSQNDRNSEHEKYNFYVNLPRGGLIVTDPDYG V.1 601DNSAVTLSILDENDDFTIDSQTGVIRPNISFDREKQESYTFYVKAEDGGRVSRSSSAXVT60 V.3 601DNSAVTLSILDENDDErIDSQTGVIRPNISFDREKQESYTFYVKAEDGGRVSRSSSAKVT 0 V.3 601 DNSAVLDNDFIDSSTGVRPSNSFDREKQSYFVKDGGRVSRSSAVT 660 V.1 661INVVDVNDNKPVFIVPPSNCSYELVLPSTNPGTVVFVIAVDNDTMNAEVRYSIVGGNT72 V.3 661INVVDVNDNKPVFIVPPSNCSYELVLPSTNPGTVVFQVIAVDNDTGMNAEVRYSIVGGNT72 V.3 6671 INVDVNADNKPVIVPPSNCSYVLPSTPVLVFQVIAVDDTGMINFEVRSVGNT 720 V.1 721RDLFAIDQETGNITLMEKCDVTDLGLHRVLVKANDLGQPDSLFSVVIVNLFVNESVTNAT78 V.3 721RDLFAIDQETGNITLEKCDVTDLGLHRVLVKANDLGQPDSLFSVVIVNLFVNESVTNAT78 V.3 721 LIDQVRSEGNITECDVDLGLHRVLVADLGPDSLFVVINLFVESTNAT 840 V.1 781LINELVRKSTEAPVTPNTEIADVSSE'TSDYVKILVAAVAGTITVVVVIFITAVVRCRQAP84 V.3: 81LINEIJVRKSTEAP VTPNTElAD VS SPT SDYVKILVAAVAGT ITVVVVIFITAVVRCRQAP84 V.1: 841 HLKAAQKNKQNSEWATPNPEWRQNINMICKKKKKKKHSPKNLLLNFVTIEETKADDVDSDG 900 Table UI(c). Nucleotide sequence of transcript variant 109PI D4 v.4 (SEQ ID NO: 249) ctggtggtcc agtacctcca aagatatgga atacactcct gaaatatcct gaaaactttt ttttttcaga atcctttaat aagcagttat gtcaatctga aagttgctta cttgtacttt 120 atattaatag ctattcttgt ttttcttatc caaagaaaaa tcctctaatc cccttttcac 180 atgatagttg ttaccatgtt taggcattag tcacatcaac ccctctcctc tcccaaactt 240 ctcttcttca aatcaaactt tattagtccc tcctttataa tgattccttg cctcgtttta 300 tccagatcaa ttttttttca ctttgatgcc cagagctgaa gaaatggact actgtataaa 360 ttattcattg ccaagagaat aattgcattt taaacccata ttataacaaa gaataatgat 420 tatattttgt gatttgtaac aaataccctt tattttccct taactattga attaaatatt 480 ttaattattt gtattctctt taactatctt ggtatattaa agtattatct tttatatatt 540 tatcaatggt ggacactttt ataggtactc tgtgtcattt ttgatactgt aggtatctta 600 tttcatttat ctttattctt aatgtacgaa ttcataatat ttgattcaga acaaatttat 660 cactaattaa cagagtgtca attatgctaa catctcattt actqatttta atttaaaaca 720 gtttttgtta acatgcatgt ttagggttgg cttcttaata atttcttctt cctcttctct 780 00 00 ctctcctctt ctggtatgga tccactctgg tgataggcga ctgctatgca aggatactgg gtatcccaag tatttagact tcccagcaac ctctcccagc t taagagtca cacaactgat taaaggttga ttactgatac cagaaaatgc gtgaaaatgc tatttcacct aaacaccaaa caatggtgct acatcgtcaa aaattgctct tcacagatca agactgcagc cagatgctgg a a aat gacaa actctcctgg ctaagatcaa caggcatgct caattctggc gcattattga tcccagaaaa atggagacaa attcacaaac acactttcta taaccataaa actgttctta ttgctgttga acacaagaga gtgatgttac ctgattctct ctacactgat agatagctga ctggcaccat caccacacct cagaaaacag agaacttgct atggaaacag acaattgggt acaaatctgc agcaccacat ccaagtgttc cgaccttcga cttgcacccc cccagcggcg gactgggaga atcctcagga actccgatcc aaccaactgt ctgatgcctg ctgctctatg tgacacagac acagcccacc cttttggtca cttgttgtcc cgcccaggag cttgttgaaa gttcaagcta tgagatcttc ggatgagcat ggttaagata agttatcaac ggctgttgat aaacattttt tgttcaaaag agatggtggc aaatgacaac tcctgtaggc caagatccac caatgccacc ccacaagtta ggtaaatgtt tcctgtcaat cataactgtg tgaaatccct atatcttgac caaacctcct tgctccaqtt catccagttg ttacctgcta gactgtagtg aaaagataac tcagaatgac ccttccaagg ttctgcagtt tggtgtcatc tgtaaaggct tgtggttgat tgaattggtt caatgacact tctgtttgca agaccttggt cttcagtgtt taatgaactg tgtatcctca aactgtcgtt taaggctgct gcagatgata gcttaatttt agtcacacta aactacacct ct ctccacag catccaagaa ctcaagcagt ggtacctgtg catgaaagag tgtcacattt ccatgatgca ggagtacttt tgaatctact ggaagaggcc ctggatgccg ccacagccca cattgctctc accgatacag gtgttgtgcg gggacgtaca aaaaactaca gaccttaact gtgtacaaga actactggcg tgcttttatg cgttttctga atatcaattc cctgacgtag ggcctcgatg gagttagata tttcctcaaa cacccagtct acttcagtga ttctctttca actggactta ctggttttgg acagatgtca gacacagttg acggataagg ttcagattaa tatgagtcca ttgaatcagt ttcacccagt acgaaagtaa ggccctgatg aagaaactag ggggtaccac aatagcccag catggtacag acgctctcca cgaccaaata gaggatggtg gtcaatgaca ctaccgtcca ggcatgaatg atcgaccaag ttacacagag gtaattgtca gtgcgcaaaa ccaactagtg gtagttattt cagaaaaaca atgatgaaga gtcactattg gaccttccta actactttca cctgccttcc ctgcctctcg tcagatccct tccgtacaca tctacaacta cacctgccag ggcagcctta gatcgtgcta ttcatacctg tctgacaact gcatctctgg ccactgtcac tgccacagcc gtgtctgctcI ggttaataca ttttcqcgqt ccatccgaga tgtcgctgat ccggagatgt ct cgcattga aagtggaggt tagaagatat cagagaactc gaataaacgg tcattgaaac gggaagagaa gatccagtac ttaaggagac cacagctcca gcaatctagt tcacaatcaa caagtgatgg atgataatgt ttctttcaga atgcggacca ggccagtatt caaaagaata cagcaatgct ctttcgtaac gtgcaatgga ctccacctga atagagaaaa ccttaaccag ttttcactca taggactaat ttttagatga tttcatttga gtagagtatc acaaaccagt ctaatccagg cagaggttcg aaacaggcaa tgttggtcaa atctgttcgt gcactgaagc actatgtcaa tcatcactgc agcagaattc aaaagaaaaa aagaaactaa ttgatctaga agcccgacag aaattcagcc ataacacctt acagcgtttc ccagaccgcc tggagatctg aaggctctca ccagcacatc cacccagcaa gactaaagaa gcactcaaga atcattccag aggcctctac ctccagtgac tccaccacag acaaactgta cctgctagca agaaatgcca tccaaacaag gccactgatt tcgtgagaaa tgccattttg aaatgataat ggctataaac agttcaaaac accagaagga ggatacctac tgctattttg agagattgaa tgccacagat ctccaacatt agaaccactg tggattgatg cccatccatt aaatattcca taatggcagg cagtaatcag tgccattaaa cttcatcaaa tgtttctatt tgcagacagt attcagcctg agaqgataaa caatgtcaca caatgaatac cactgtaact gaatgatgac tagagaaaaa acgttcttca tttcattgtc cacagtggtc ttacagcatt cataacattg agctaatgac gaatgagtcg accagtgacc gatcctggtt tgtagtaaga tgaatgggct gaagaagaag ggcagatgat agagcaaaca ccctgatttg tgaaactccc tgtggcctgt tgactgtggc aatgaaggag gattcatccc ggaaagcagc tcatggcctg tcgcactgaa agctgcagaa atgtctcatc ctcttcgcaa tcagcaccac acagaccatc tcctcctcta acaagtqtac tgcgtggtgt gaaaacgtcc tccttgacaa cgaattgaag ttatgtgctg ccggatgaaa gcaccattgt tctaaatata tacgaactaa gacaagatgc gtgatgaaag caagtgagtg gtcagtatac gctgacatag gccaggagat gatagggaag ccagcaagag gacataagat ctcaacacca gtgacatgct ttcctcctgg ttactggctg gtgaaagatg cctgagaata gggcctaatg gattgtcgta tatttattca gtctttgtaa aacttctatg gatcctgatt ttcaccattg caagaatctt agtgccaaag cctccttcca tttcaggtaa gtaggaggaa atggagaaat ttaggacagc gtgaccaatg ccaaatactg gcagctgttg tgtcgccagg accccaaacc cattccccta gttgacagtg atgggaaagt gcccgacact ctgaattcga gactctatct tatccagtga gttgtgcgat caaccacagt agtgatggtg ccccttggct ggggatggca ataactgttc tatggccatt gcacaggcct agcccacgag gcattgtgcc gtgcaggcta 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 00 00 ctgcacttca tagcacaggc gtagtcaggc tatgctctgt aaagacttca gccaacaggc gctaaaatag ttccaatgag agaataaatc attcagaatg aggcttatca tcatgtgcaa tggaaagcct cagataacgt gtttgttgtt gtagtgtaaa actatattgt agatctagat ataactgccc agtgtaaagt acttgggtct atgcatccaa ctcaggaaga taaaaataac gqatttttgt aggaaggaac aatgcagagg acacacacag caaagaaaca cttgtatcct atcaccaact ctaaaccaag tctcctattg attaaactgt ggatttccac ttcagaatat tgcagaataa ttttctcctt atgtgaatcc tgttgatatt gaaccaatag aaacctgcaa ctgcaaaact acacacattt atctcaatgg ggatttttct tctggaaact gcttttatta taaatgactg gagagtagcc ctttctaagc gatgatatct gttgacacag aacgtcttac ttgaaattaa ttattctatt ttactatatc tacacat tct aattagcttt cttatttact ggttagtgct tttatcacac ccacagccca tgctgcaatc ccaatcatca tgatcaggga tcccagtgat cagaccgtcc ttacttcaaa tattctgatt tacagctaga tgtatttaaa tgacagagcg tattactgat agaaatatct taaaaggtat ttcagttttt tactgcttgt tgataaaatt ctacaaccta taattaagca acatcagaaa ttacttctgg agcacgagtc tttaaatgtt aaatacatgt tattctagct ttttcaagaa gagaaataag aacaaaaacc gtactattca cttaactggc taattttctt agacaattga attaacttag gtgtaaattt tgaatattga acattattcc cttcttaact cgtgttgtta cgaaggactg gtggctattg tgacaaaaat agatgatgtg ttagttttct aaatgatata ttactatcta tctgacagta tgtgacgttt ccctttctaa ggcaccctct aaagaggcaa gtccactagg ttttcatcac cacacaagtg tgaaccaaca aacttcaagc ttgtgtccat acacttgtga tttcttaaac tacatatgat taacatttta attgtgtaat at ggt cat ta ccatcagcac agccacagct gtcagtttgc gtgcaaggta gattcaatta agaggtgatt ttttcagaaa atcagatttg cccttagtca aagaaaagga cactatttct ttgtttccat tattttctaa acgtactcta ttgttgttgt ttgtgtctct tgtatataca tttctctact actatttgtt taaagctgta gaatttgtat acttaaaata gatttgacag aactgtagat caacttactg atgtaattat gcacatgact atcaaaatct tagaaaacat cattatcttg ccatagcaaa tgtttaatgg acatagattt tgtaacataa qaaagtttct catccattgt tttaactatc atgttccaag atgatatttg ttgatgttga cagtgcttcc agattttttc gatgatctac aaaagaagag tgttagtgaa taatttattg tgacaccttt ctctgatttt tcttggtttt ggggtattag tctggccttg attacaagtt gctattgtga atcaaaaaat tgaatgactt attccattag gtatgtattc tttacctgtg atctacaatg cttttaatta ggtagcagtt aatgggaaaa aggcctcagc ctcctctgcc agcaaggttg gtgcaacatc aagtcattcc cccccattat agatgtatat taaataacta atagttaacc atttaacaat gatgtacagt gctgattgtg gtttaccttt gccttttttt tagtgagtct ctgctgtcat ttttcaataa ctttagtaga aaaaagggcc tctgccattt gtaacagcct tcgacggtaa ctcaataggc aaaaccatat aagaaaacca aaatctacat gcttcttgca catatatqaa tagttttctt tatgtgcaca actgagaaaa gggcggttgg tgtaatgtat actgtggtaa tttcatgtqc aaagttcctt agagtttgat ggatttggag aatgtttatt aaattttaaa agtagatttt ttgtgtttta acacacacac gttgaaagat aatcaaatag agttactagg cctatagatg ataatcagac taccagagag cccagttatt gaaacctgtt atttctctga tgtatgatgt ggttctgttt atatgagaat attgtgatta aaatactaag ttttaactaa taataaattt tgtaaatttg acaaagagcc aaatagacta cctctgctac acaggttatt ggtgcaaggt tcagttttac tttgacaacc ggaagaacat agtcaaaatt tgtaaataga aaaaaattgc ttgcatcccc attttttgtt tggaaccagt agtttaccta gggctttctt cccttcaaaa gttttctacc agaatatgta gttcgagaca tctttttact taagcctgta agaaaattaa actactattt tgttaccaaa actaaatcta ctaataacaa caaacagaat gtcaacaaga ataaaatata ctgttgtctg ttttataaat taccttgttt ggtggggggg aacttgatat ttgcataatt ccagcaggtt aagtcatatt taataaaatt catactggtt aaattattat cttggggaag agaacattct attattttca acacacacac tattaaataa gactcaaagt gaggttctta atataggaat ttagattgtg gctttgaatg ctcccctatg acttctaggg ctgaatagac atgtagtcct taaaaaggat aatacgttca ttaattttct tatcttatat tattgtgtca agagagtaat gttagaaaat tctgccttcc aacaaatcac aqccctcctt gccctccatc gctgatgggc accatgtctg ttcactccac cccttgtaaa taagatacaa aacagatacc aatttgttta ttgtacagta gtttttatca atgtagcaaa aacttttgtt tttgattttt tacgcagtag ttattccaat taaactgtac cagaagtgca ttaatagttt gtccattatt aaggaggtgg tgtagagaaa gggtgttcag taagactaag caagaatatc tttaaggaaa aataccaata ttcttctaag ttatttcctt gtacagaaac cagtataaca ggagtcaata ttaatttatg tcattggtga aagtagcgtt tgactgggcg aattaatgtt ttccaggtgc catacaaatg attaagaaaa ttgcctcaaa cattttctct gtgcacacac cttatcaggc tggatatttg aatcctcata gaaccaatac tttagaatat gaagcaggct ccttccttct cttcagatct agtggtatag acaactgcaa tttgtttgat atcaaagtag agctatggta gctacgtgca gtgtattaaa tttgtgtatt aataataaat caaactaata aaattgttca 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 51160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 5940 6000 6060 6120 6180 6240 6300 6360 6420 6480 6540 6600 6660 6720 6780 6840 6900 6960 7020 7080 7140 7200 7260 7320 7380 7440 7500 7560 7620 7680 7740 7800 7860 7920 7980 8040 8100 8160 8220 00 00 gttcttaaaa tgtaattatg tcacacacac aaaaaatcct tttcaatcct aaggcgtttt actcacatgg ctatttcaac attagttttt tttgtttgtt ggtattactg aaggtgtgta tactccctaa tacacattta tgaaaatcta cttttattta tactcttctg atttatattt tttattataa ttattatttc tcttttatat tttttqgaaa ccaaatttat agttagttta ggtaaacttt cattagaaac tattttgaat gcttccaact ggctcaattg gccgggaaaa agagaagctg aaatatattt ctgcaagaac ctttctatat tatgtgccaa agatcaattt tatgcagagg ccttaaaata ttctttcaca gtagctttct gtcatgtgct tttagtaaat atgattttta aaagcagttc aagttgacaa agtaacaaaa aaatctgctc agaaaaatgt atgtgcacaa ataaaaaaaa ttgtttagtg attgtaagtg atacttttta aagagtaaac tgtgtgaaat cctgcttaaa atattaagat ttttatgaaa tatgtattta tgtttgtatt tcctcctctg tgatatcata cagcatctga aagtgaacag tatcccaaag catgctttgg aagtaagaag gttgactatt gtatggccaa ggatggcagt gaagcaaact tgtattaatt gttctatttc aggttctgta ttgcatgttt atatattaat aaaagttatg agaaat gagaaaatta tctttttcat cttgtttagg ttatctttct ttattatgac catgggagca ttaccacacc tacactaac cagcagaaac ttaatqgcaa ttatactatc gtgggaagat cagttccaac atgtaatcca tcttattaat 8280 8340 8400 8460 8520 8580 8640 8700 8760 8820 8880 8940 9000 9060 9120 9146 Table UII(c). Nucleotide sequence alignment of 109P1 D4 v.1 (SEQ ID NO: 250) and i 09P1 D4 v.4 (SEQ ID NO: 251) Score 7456 bits (3878), Expect =0.Oldentites =3878/3878 (100%) Strand =Plus Plus V. 1 1 ctggtggtccagtacctccaaagatatggaatacactcctgaaatatcctgaaaactttt V.4 1 ctggtggtccagtacctccaaagatatggaatacactcctgaaatatcctgaaaactttt V.1 61 ttttttcagaatcctttaataagcagttatgtcaatctgaaagttgcttacttgtacttt 120 VA4 61 ttttttcagaatcctttaataagcagttatgtcaatctgaaagttgcttacttgtacttt 120 V.1 121 atattaatagctattcttgtttttcttatccaaagaaaaatcctctaatccccttttcac 180 V.4 121 atattaatagctattcttgtttttcttatccaaagaaaaatcctctaatccccttttcac 180 V.1 181 atgatagttgttaccatgtttaggcattagtcacatcaacccctctcctctccaaactt 240 V.4A 181 atgatagttgttaccatgtttaggcattagtcacatcaacccctctcctctcccaaactt 240 V.1 241 ctcttcttcaaatcaaactttattagtccctcctttataatgatttcttgcctcgtttta 300 V.A 241 ctcttcttcaaatcaaactttattagtccctcctttataatgattccttgcctcgtttta 300 V.1 301 tccagatcaattttttttcactttgatgcccagagctgaagaaatggactactgtataaa 360 VA4 301 tccagatcaattttttttcactttgatgcccagagctgaagaaatggactactgtataaa 360 V.1 361 ttattcattgccaagagaataattgcattttaaacccatattataacaaagaataatgat 420 VA. 361 ttattcattgccaagagaataattgcattttaaacccatattataacaaagaataatgat 420 V.1 :421 tatattttgtgatttgtaacaaataccctttattttcccttaactattgaattaaatatt 480 VA.4 421 tatattttgtgatttgtaacaaataccctttattttcccttaactattgaattaaatatt 480 V.1 :481 ttaattatttgtattctctttaactatcttggtatattaaagtattatcttttatatatt 540 V. 481 ttaattatttgtattctctttaactatcttggtatattaaagtattatcttttatatatt 540 V.1 :541 tatcaatggtggacacttttataggtactctgtgtcatttttgatactgtaggtatctta 600 260 0V.4 541 tatcaatggtggacattttataggtactctgtgtcatttttgatactgtaggtatctta 600 V.1 601 tttcatttatctttattcttaatgtacgaattcataatatttgattcagaacaaatttat 660 V.4 601 tttcatttatctttattcttaatgtacgaattcataatatttgattcagaacaaatttat 660 tnV.1 661 cactaattaacagagtgtcaattatgctaacatctcatttactgattttaatttaaaaca 720 VA4 661 cactaattaacagagtgtcaattatgctaacatctcatttactgattttaatttaaaaca 720 V.1 721 gtttttgttaacatgcatgtttagggttggcttcttaataatttcttcttcctcttctct 780 V.4 721 gtttttgttaacatgcatgtttagggttggcttcttaataatttcttcttcctcttctct 780 CN1V.1 781 ctctcctcttcttttggtcagtgttgtgcgggttaatacaacaaactgtaacaagtgtac 840 V.4 781 ctctcctcttcttttggtcagtgttgtgcgggttaatacaacaaactgtaacaagtgtac 840 V.1 841 ctggtatggacttgttgtccgggacgtacattttcgcggtcctgctagcatgcgtggtgt 900 V.4 841 ctggtatggacttgttgtccgggacgtacattttcgcggtcctgctagcatgcgtggtgt 900 V.1 901 tccactctggcgcccaggagaaaaactacaccatccgagaagaaatgccagaaaacgtcc 960 V.4 901 tccactctggcgcccaggagaaaaactacaccatccgagaagaaatgccagaaaacgtcc 960 V.1 :961 tgataggcgacttgttgaaagaccttaacttgtcgctgattccaaacaagtccttgacaa 1020 VA.4 961 tgataggcgacttgttgaaagaccttaacttgtcgctgattccaaacaagtccttgacaa 1020 V.1 :1021 ctgctatgcagttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaag 10B0 VA.4 1021 ctgctatgcagttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaag 1080 V.1 :1081 aggatactggtgagatcttcactactggcgctcgcattgatcgtgagaaattatgtgctg 1140 VA.4 1081 aggatactggtgagatcttcactactggcgctcgcattgatcgtgagaaattatgtgctg 1140 V.1: 1141 gtatcccaagggatgagcattgcttttatgaagtggaggttgccattttgccggatgaaa 1200 VA 1141 gtatcccaagggatgagcattgcttttatgaagtggaggttgccattttgccggatgaaa 1200 V.1 1201 tatttagactggttaagatacgttttctgatagaagatataaatgataatgcaccattgt 1260 VA4 1201 tatttagactggttaagatacgttttctgatagaagatataaatgataatgcaccattgt 1260 V.1 1261 tcccagcaacagttatcaacatatcaattccagagaactcggctataaactctaaatata 1320 VA4 1261 tcccagcaacagttatcaacatatcaattccagagaactcggctataaactctaaatata 1320 V.1 1321 ctctcccagcggctgttgatcctgacgtaggaataaacggagttcaaaactacgaactaa 1380 VA4 1321 ctctcccagcggctgttgatcctgacgtaggaataaacggagttcaaaactacgaactaa 1380 V.1 1381 ttaagagtcaaaacatttttggctcgatgtcattgaaacaccagaaggagacaagatgc 1440 261 00 111 iiI1111jl 111111I111lIl1

I

VA4 1381 ttaagagtcaaaacatttttggcctcgatgtcattgaaacaccagaaggagacaagatgc 1440 V.1 1441 cacaactgattgttcaaaaggagttagatagggaagagaaggatacctacgtgatgaaag 1500 V.4 1441 cacaactgattgttcaaaaggagttagatagggaagagaaggatacctacgtgatgaaag 1500 V.1 1501 taaaggttgaagatggtggctttcctcaaagatccagtactgctattttgcaagtgagtg 1560 V.4 1501 taaaggttgaagatggtggctttcctcaaagatccagtactgctattttgcaagtgagtg 1560 C1V.1 1561 ttactgatacaaatgacaaccacccagtctttaaggagacagagattgaagtcagtatac 1620 C1V.4 1561 ttactgatacaaatgacaaccacccagtctttaaggagacagagattgaagtcagtatac 1620 00 V.1 1621 cagaaaatgctcctgtaggcacttcagtgacacagctccatgccacagatgctgacatag 1680 V.4 1621 cagaaaatgctcctgtaggcacttcagtgacacagctccatgccacagatgctgacatag 1680 V.1 1681 gtgaaaatgccaagatccacttctctttcagcaatctagtctccaacattgccaggagat 1740 VA4 1681 gtgaaaatgccaagatccacttctctttcagcaatctagtctccaacattgccaggagat 1740 V.1 1741 tatttcacctcaatgccaccactggacttatcacaatcaaagaaccactggatagggaag 1800 VA4 1741 tatttcacctcaatgccaccactggacttatcacaatcaaagaaccactggatagggaag 1800 V.1 1801 aaacaccaaaccacaagttactggttttggcaagtgatggtggattgatgccagcaagag 1860 VA4 1801 aaacaccaaaccacaagttactggttttggcaagtgatggtggattgatgccagcaagag 1860 V.1 1861 caatggtgctggtaaatgttacagatgtcaatgataatgtcccatccattgacataagat 1920 VA4 1861 caatggtgctggtaaatgttacagatgtcaatgataatgtcccatccattgacataagat 1920 V.1 1921 acatcgtcaatcctgtcaatgacacagttgttctttcagaaaatattccactcaacacca 1980 VA4 1921 acatcgtcaatcctgtcaatgacacagttgttctttcagaaaatattccactcaacacca 1980 V.1 1981 aaattgctctcataactgtgacggataaggatgcggaccataatggcagggtgacatgct 2040 VA4 1981 aaattgctctcataactgtgacggataaggatgcggaccataatggcagggtgacatgct 2040 V.1 2041 tcacagatcatgaaatccctttcagattaaggccagtattcagtaatcagttcctcctgg 2100 VA4 2041 tcacagatcatgaaatccctttcagattaaggccagtattcagtaatcagttcctcctgg 2100 V.1 2101 agactgcagcatatcttgactatgagtccacaaaagaatatgccattaaattactggctg 2160 VA4 2101 agactgcagcatatcttgactatgagtccacaaaagaatatgccattaaattactggctg 2160 V.1 2161 cagatgctggcaaacctcctttgaatcagtcagcaatgctcttcatcaaagtgaaagatg 2220 VA4 2161 cagatgctggcaaacctcctttgaatcagtcagcaatgctcttcatcaaagtgaaagatg 2220 262 00 V.1 2221 aaaatgacaatgctccagttttcacccagtctttcgtaactgtttctattcctgagaata 2280 V.4 2221 aaaatgacaatgctccagttttcacccagtctttcgtaactgtttctattcctgagaata 2280 V.1 2281 actctcctggcatccagttgacgaaagtaagtgcaatggatgcagacagtgggcctaatg 2340 V.4 2281 actctcctggcatccagttgacgaaagtaagtgcaatggatgcagacagtgggcctaatg 2340 V.1 2341 ctaagatcaattacctgctaggccctgatgctccacctgaattcagcctggattgtcgta 2400 V.4 2341 ctaagatcaattacctgctaggccctgatgctccacctgaattcagcctggattgtcgta 2400 V. 41cgctctattggaaataaaaaaaagtaaatata26 V.1 2401 caggcatgctgactgtagtgaagaaactagatagagaaaaagaggataaatatttattca 2460 00 V.1 2461 caattctggcaaaagataacggggtaccacccttaaccagcaatgtcacagtctttgtaa 2520 (iV.4 2461 caattctggcaaaagataacggggtaccacccttaaccagcaatgtcacagtctttgtaa 2520 V.1 2521 gcattattgatcagaatgacaatagcccagttttcactcacaatgaatacaacttctatg 2580 VA4 2521 gcattattgatcagaatgacaatagcccagttttcactcacaatgaatacaacttctatg 2580 V.1 2581 tcccagaaaaccttccaaggcatggtacagtaggactaatcactgtaactgatcctgatt 2640 VA4 2581 tcccagaaaaccttccaaggcatggtacagtaggactaatcactgtaactgatcctgatt 2640 V.1 2641 atggagacaattctgcagttacgctctccattttagatgagaatgatgacttcaccattg 2700 VA4 2641 atggagacaattctgcagttacgctctccattttagatgagaatgatgacttcaccattg 2700 V.1 2701 attcacaaactggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatctt 2760 VA4 2701 attcacaaactggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatctt 2760 V.1 2761 acactttctatgtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaag 2820 VA4 2761 acactttctatgtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaag 2820 V.1 2821 taaccataaatgtggttgatgtcaatgacaacaaaccagttttcattgtccctccttcca 2880 VA.4 2821 taaccataaatgtggttgatgtcaatgacaacaaaccagttttcattgtccctccttcca 2880 V.1: 2881 actgttcttatgaattggttctaccgtccactaatccaggcacagtggtctttcaggtaa 2940 VA. 2881 actgttcttatgaattggttctaccgtccactaatccaggcacagtggtctttcaggtaa 2940 V.1 :2941 ttgctgttgacaatgacactggcatgaatgcagaggttcgttacagcattgtaggaggaa 3000 VA.4 2941 ttgctgttgacaatgacactggcatgaatgcagaggttcgttacagcattgtaggaggaa 3000 V.1 :3001 acacaagagatctgtttgcaatcgaccaagaaacaggcaacataacattgatggagaaat 3060 VA.4 3001 acacaagagatctgtttgcaatcgaccaagaaacaggcaacataacattgatggagaaat 3060 00 V.1 3061 gtgatgttacagaccttggtttacacagagtgttggtcaaagctaatgacttaggacagc 3120 V.4 3061 gtgatgttacagaccttggtttacacagagtgttggtcaaagctaatgacttaggacagc 3120 V.1 3121 ctgattctctcttcagtgttgtaattgtcaatctgttcgtgaatgagtcggtgaccaatg 3180 V.4 3121 ctgattctctcttcagtgttgtaattgtcaatctgttcgtgaatgagtcggtgaccaatg 3180 V.1 31B1 ctacactgattaatgaactggtgcgcaaaagcactgaagcaccagtgaccccaaatactg 3240 V.4 3181 ctacactgattaatgaactggtgcgcaaaagcactgaagcaccagtgaccccaaatactg 3240 V. 21aaacgagactacatgtatttagtcggtcgttg30 3241 agatagctgatgtatcctcaccaactagtgactatgtcaagatcctggttgcagctgttg 3300 00 V.1 3301 ctggcaccataactgtcgttgtagttattttcatcactgctgtagtaagatgtcgccagg 3360 V.4 3301 ctggcaccataactgtcgttgtagttattttcatcactgctgtagtaagatgtcgccagg 3360 V.1 3361 caccacaccttaaggctgctcagaaaaacaagcagaattctgaatgggctaccccaaacc 3420 V.A 3361 caccacaccttaaggctgctcagaaaaacaagcagaattctgaatgggctaccccaaacc 3420 V.1 3421 cagaaaacaggcagatgataatgatgaagaaaaagaaaaagaagaagaagcattccccta 3480 V.4 3421 cagaaaacaggcagatgataatgatgaagaaaaagaaaaagaagaagaagcattccccta 3480 V.1 3481 agaacttgctgcttaattttgtcactattgaagaaactaaggcagatgatgttgacagtg 3540 V.4 3481 agaacttgctgcttaattttgtcactattgaagaaactaaggcagatgatgttgacagtg 3540 V.1 3541 atggaaacagagtcacactagaccttcctattgatctagaagagcaaacaatgggaaagt 3600 VA4 3541 atggaaacagagtcacactagaccttcctattgatctagaagagcaaacaatgggaaagt 3600 V.1 3601 acaattgggtaactacacctactactttcaagcccgacagccctgatttggcccgacact 3660 VA4 3601 acaattgggtaactacacctactactttcaagcccgacagccctgatttggcccgacact 3660 V.1 3661 acaaatctgcctctccacagcctgccttccaaattcagcctgaaactcccctgaattcga 3720 VA4 3661 acaaatctgcctctccacagcctgccttccaaattcagcctgaaactcccctgaattcga 3720 V.1 3721 agcaccacatcatccaagaactgcctctcgataacacctttgtggcctgtgactctatct 3780 VA4 3721 agcaccacatcatccaagaactgcctctcgataacacctttgtggcctgtgactctatct 3780 V.1 3781 ccaagtgttcctcaagcagttcagatccctacagcgtttctgactgtggctatccagtga 3840 VA4 3761 ccaagtgttcctcaagcagttcagatccctacagcgtttctgactgtggctatccagtga 3840 V.1 3841 cgaccttcgaggtacctgtgtccgtacacaccagaccg 3878 VA4 3841 cgaccttcgaggtacctgtgtccgtacacaccagaccg 3878 00 0 0 ci

C)

Co 0 Table UV(c). Peptide sequences of protein coded by 1091D4 v.4 (SEQI ID NO: 252) MDLLSGTYIF AVLLACVVFH SGAQEKNYTI REEMPENVLI GDLLKDLNLS MQFKLVYKTG DVPLIRIEED TGEIFTTGAR IDREKLCAGI PRDEHCFYEV RJJVKIRFLIE DINDNAPLFP ATVINISIPE NSAINSKYTL PAAVDPDVGI SQNIFGLDvI ETPEGDKNPQ LIVQKELDRE EKDTYVMKVK VEDGGFPQRS DTNDNHPVFK ETEIEVSIPE NAPVGTSVTQ LHATDADIGE NAKIHFSFSN JILNATTGLIT IKEPLDREET PNHKrJLVLAS DGGLMPARAM VLVNVTDVND VNPVNDTVVL SENIPLNTKI ALITVTDKDA DHNGRVTCFT DHEIPFRLRP AAYLDYESTK EYAIKLLAAD AGKPPLNQSA MLFIKVKDEN DNAPVFTQSF PGIQLTKVSA MDADSGPNAK INYLLGPDAP PEFSLDCRTG MLTVVKKLDR LAKDNGVPPL TSNVTVFVSI IDQNDNSPVF THNEYNFYVP ENLPRHGTVG DNSAVTLSIL DENDDFTIDS QTGVIRPNIS FDREKQESYT FYVKAEDGGR INVVDVNDNK PVFIVPP.SNC SYELVLP.STN PGTVVFQVIA VDNDTGMNAE RDLFAIDQET GNITLMEKCD VTDLGLHRVL VKANDIJGQPD SIJFSVVIVNL LINELVRKST EAPVTPNTEI ADVSSPTSDY VKILVAAVAG TITVVVVIFI MLKAAQKNKQ NSEWATPNPE NflQMIMMKKK KKKKKHSPKN LLLNFVTIEE NRVTLDLPID LEEQTI4GKYN WVTTPTTFKP DSPDLARHYK SASPQPAFQI HIIQEIJPLDN TFVACDSISK CSSSSSDPYS VSDCGYPVTT FEVPVSVHTR TPNKESTTME IWIHPQPQSQ RRVTFHLPEG SQESSSDGGL GDHDAGSLTS QEEYFDRATP SNRTEGDGNS DPESTFIPGL KKAAEITVQP TVEEASDNCT ACWMPASLDH SSSSQAQASA LCHSPPLSQA STQHHSPRVT QTIALCHSPP PPEIQVSALH HSPPLVQATA LHHSPE'SAQA SAIICYSPPLA OAAAISHSSP QAQSSVSLQQ GWVQGADGLC SVDQGVQGSA TSQFYTMSER LMFSDDSIKV QARPSRGDSP IMEEHPL

LIPNKSLTTA

EVAILPDEIF

NGVQNYELIK

S TM LOVS VT

LVSNIARRLF

NVPSIDIRYI

VFSNQFLLET

VTVSI PENNS

EKEDKYLFTI

LITVTDPDYG

VSRSSSAKVT

VRYS IVGGNT

FVNESVTNAT

TAVVRCRQAP

TKADDVDSDG

QPETPLNSKH

PPMKEVVRSC

TSHGLPLGYP

QECLIYGHSD

VTQTIALCHS

LPQVIALHRS

I PLTTFTPRQ 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1337 Table LV(c). Amino acid sequence alignment of 109P1 D4 v.1 (SEQ ID NO: 253) and 109P1 D4 v.4 (SEQ ID NO: 254) Score =2005 bits (5195), Expect 0.Oldenfifles 10111/1011 Posities =1011 /1011 (100%) V.1 1 I4DLLSGTYIFAVLIACVVFHSGAQEK14YTII1EEMPENVLIGDLIJ(DWLSLIPNKSJTTA MDLLSGTYI FAVLLACVVFHSGAOEKNYTIREHPENVLIGDLLKDLNLSLIPNKSLTTA V.4 1 MDLLSGTYIFAVLLACVVFHSGAQEEQYTIREEMPENVLIGDLLKDLNLSLIPNKSLTTA V.1 61 M0FKLVYKTGDVPLIRIEEDTCEIFTTGAJRIDREKLCAGIPRDEHCFYEVEVMILPDEIF 120 HOFKLVYKTGDVPLIRIEEDTGEIFrTGARIDREKLCAGIPRDEHCFYEVEVAILPDEIF V.4 61 MQKVKGVLREDGlrGRIRKCGPDHFEEALDI 120 V.1 121 RLVKIRFLIEDINDNAPLFPATVINISIPENSAIWSKYTLPAAVDPDVGINGVQNYELIK 180

RLVKIRFLIEDINDWAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVQNYELIK

V.4 121 RLVKIRFLIEDINDNAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVQNYELIK 180 V.1 181 SONIFGIJDVIETPEGDKMPQLIVQKELDREEICOTYVIKVKVEDGGFPQRSSTAILQVSVT 240

SONIFGLDVIETPEGDKMPQLIVQKIJDREEKDTYVMCVXVEDGGFPQRSSTAILQVSVT

V.4 181 SQNIFGLDVIETPEGDKMPQLIVQKELDREEKDTYVMKVKVEDGGFPQRSSTAILQVSVT 240 V.1 241 DTNDNHPVFKETEIEVSIPENAPVGTSVTQLHATDADIGENJCIHFSFNLVSNIARRU.IF 300 VA4 241 DTNDNHPVFKETEIEVSIPENAVGTSVTQLATDADIGEN(IHFSFSNLVSIAPJF 300 V.1 301 HLNATTGLITIKEPLDREETPNHKILVLASDGGLMPAnI4VLVNVTDVNDNVPSIDIRYI 360

HLNATTGLITIKEPLDREETPNHKLLVLASDGGLMPAAMVLVNVTDVNDNVPSIDIRYI

VA4 301 HLNATTGLITIKEPLDREETPNHJCILVLASDGGLMPA3ANVLVNWDVNqDNVPsIDIRYI 360 V.1 361 VNFVNDTWVLSENIPLNTKIALITVTDKDADHNGRVTCFTDHEIPFRLRPVFSNQFLLET 420 VNPVNDTVVLSENIPLNTKIALITVTDKDADHNGRVTCrTDHEIPFRLRPVFSNQFLLET VA.4 361 VNPVNDTVVLSENIPLNTKIALITVTDKDADHNGRVTCrDHEI PFRLRPVFSNQFLLET 420 V.1 :421 AAYLDYESTKEYAIKLLAADAGKPPLNQSA4IFIKVKDENDNAVQS FVTVVI PENNqS 480

AAYLDYESTKEYAIKLLAADAGKPPLNOSAMLFIKVKDENDNAPVFTQSFVTVSIPENNS

VA.4 421 AAYLDYESTKEYAIKLLADAGKPPLNQSAMLFIKKDENDNAPVFTQspy'VTSXpENNjs 480 V.1 :481 PGIQLTKVSAMDADSGPNAJCINYLLGPDAPPEFSLDCRTGHLTVVKXLDREKEDI{YLTI 540 PGIQLTKVSAMDADSGPNAINYLLGPDAPPEFSLDCRTG4LTVVCXLDREKEDKY7JFTI 00 V.A 481 PGIQLTKVSAIMDADSGPNAKINYLLGPDAPPEFSLDCRTGMLTVVKKLDREKEDKYLFTI 540 V.1 541 LAKDNGVPPLTSNVTVFVSIIDQNDNSPVF'rHNEYNFYvpENLPRHGTVGLITVTDPDYG 600 LAKDNGVPPLTSNVTVFVSIIDQNDNoTNsv.EYtqFYVPELPRHGTVGLITVTDPDYG V.4 541 LAKDNGVPPLTSNVTVFVSIIDQNDNSPVFTHiNEYNFYVPE4LPRHGTVGLITVTDPDYG 600 V.1 601 DNSAV'LSILDENDDFTIDSQTGVIRPNISFDREKQESYTFYVKAEDGGRVSRSSSAKVT 660

DNSAVTLSILDENDDFTIDSQTGVIRPNISFDREKQESYTFYVKAEDGGRVSRSSSACVT

V.4 601 DNSAVTLSILDENDDFTIDSQTGVIRPNISFDREKQESYTFYVKAEDGGRVSRSSSAKVT 660 V.1 :661 INVVDVNDNKPVFIVPPSNCSYELVLPSTNPGTVVFQVIAVDNDTGMNAEVRYSIVGGNT 720

INVVDVNDNKPVFIVPPSNCSYELVLPSTNPGTWFQVIAVDNDTGMNAEVRYSIVGGNT

VA. 661 INVVDVNDNKPVFIVPPSl4CSYELVLPSTNPGTVVFQVAVDNDTGMNAEVRYSIVGGNT 720 V.1 :721 RDLFAIDQETGNITLMEKCDVTDLGLHRVLVKANDLGQPDSLFSVVIVNLFVNESVTNAT 780 RDLFAIDQETGNITLMEKCDVTDLGLHRVLVKAd4DLGQPDSLFSVVIVNLFVNESVTNAT VA.4 721 RDLFAIDQETGNITLMEKCDVTDLGLHRVLVKANDLGQPDSLFSVVIVNLFVESVTNAT 780 V.1 :781 LINELVRKSTEAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVVVIFITAVVRCRQAP 840

LINELVRKSTEAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVVVIFITAVVRCRQAP

VA. 781 LINELVRKSTFAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVWVIFITAWVRCRQAP 840 V.1: 841 HLKAAQIK4KQNSEWATPNPENR MIMM KKKKHSPKNLLLNFVTIEETKADDVDSDG 900 HLKAAQKKQNSEWATPNPENRQMIMflKKKKXXKHSPKNLLLNFVTIEETKADDVDSDG VA. 841 HLKAAQKNKQNSEWATPNPENRQMIMM MKKKSPNLLLNFVTIETKADDVDSDG 900 V.1 :901 NRVTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLARHYKSASPQPAFQIQPETPLNSKH 960

NRVTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLARHYKSASPQPAFQIQPETPLNSKH

VA.4 901 NRVTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLARHYKSASPQPAFQIQPETPLNSKH 960 V.1: 961 HIIQELPLDNTFVACDSIS(CSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP 1011 HIIQELPLDNTFVACDSI SKCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP VA.4 961 HIIQELPLDNTEVACDSISKCSSSSSDPYSVSDCGYPVTTF7EVPVSVHTRP 1011 Table UI(d). Nucleotide sequence of transclpt variant 109P1 D4 v.5 (SEQ ID NO: 255) ctggtggtcc agtacctcca aagatatgga atacactcct gaaatatcct g ttttttcaga atcctttaat aagcagttat gtcaatctga aagttgctta c atattaatag ctattcttgt ttttcttatc caaagaaaaa tcctctaatc c atgatagttg ttaccatgtt taggcattag tcacatcaac ccctctcctc t ctcttcttca aatcaaactt tattagtccc tcctttataa tgattccttg c tccagatcaa ttattcattg tatattttgt ttaattattt tatcaatggt tttcatttat cactaattaa gtttttgtta ctctcctctt ctggtatgga tccactctgg t gataggcga ctgctatgca aggatactgg gtatcccaag tatttagact tcccagcaac ctctcccagc ttaagagtca cacaactgat taaaggttga ttactgatac cagaaaatgc gtgaaaatgc tatttcacct ttttttttca ccaagagaat gatttgtaac gtattctctt ggacactttt ctttattctt cagagtgtca acatgcatgt cttttggtca cttgttgtcc cgcccaggag cttgttgaaa gttcaagcta tgagatcttc ggatgagcat ggttaagata agttatcaac ggctgttgat aaacattttt tgttcaaaag agatggtggc aaatgacaac tcctgtaggc caagatccac caatgccacc ctttgatgcc aattgcattt aaataccctt taactatctt ataggtactc aatgtacgaa attatgctaa ttagggttgg gtgttgtgcg gggacgtaca aaaaactaca gaccttaact gtgtacaaga actactggcg tgcttttatg cgttttctga atatcaattc cctgacgtag ggcctcgatg gagttagata tttcctcaaa cacccagtct acttcagtga ttctctttca actggactta cagagctgaa taaacccata tattttccct ggtatattaa tgtgtcattt ttcataatat catctcattt cttcttaata ggttaataca ttttcgcggt ccatccgaga tgtcgctgat ccggagatgt ctcgcattga aagtggaggt tagaagatat cagagaactc gaataaacgg tcattgaaac gggaagagaa gatccagtac ttaaggagac cacagctcca gcaatctagt tcacaatcaa gaaatggact ttataacaaa taactattga agtattatct ttgatactgt ttgattcaga actgatttta atttcttctt acaaactgta cctgctagca agaaatgcca tccaaacaag gccactgatt tcgtgagaaa tgccattttg aaatgataat ggctataaac agttcaaaac accagaagga ggatacctac tgctattttg agagattgaa tgccacagat ctccaacatt agaaccactg a a t a a a

C

a t 9 t c t c g t t 9 9 c g g g g jaaaactttt :ttgtacttt :ccttttcac :cccaaactt ~ctcgtttta .ctgtataaa raataatgat ttaaatatt ttatatatt .ggtatctta .caaatttat .tttaaaaca ctcttctct .caagtgtac gcgtggtgt aaaacgtcc ccttgacaa gaattgaag tatgtgctg :cggatgaaa caccattgt ctaaatata acgaactaa 'acaagatgc tgatgaaag .aagtgagtg 'tcagtatac ctgacatag ccaggagat atagggaag 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 00 00 aaacaccaaa caatggtgct acatcgtcaa aaattgctct tcacagatca agactgcagc cagatgctgg aaaatgacaa actctcctgg ctaagatcaa caggcatgct caattctggc gcattattga tcccagaaaa atggagacaa attcacaaac acactttcta taaccataaa actgttctta ttgctgttga acacaagaga gtgatgttac ctgattctct ctacactgat agatagctga ctggcaccat caccacacct cagaaaacag agaacttgct atggaaacag acaattgggt acaaatctgc agcaccacat ccaagtgttc cgaccttcga acctgccaga gcagccttac atcgtgctac tcatacctgg ctgacaactg catctctgga cactgtcaca gccacagccc tgtctgctct catcagcaca gccacagctc tcagtttgca tgcaaggtag attcaattaa gaggtgattc tttcagaaaa tcagatttgt ccttagtcaa agaaaaggaa actatttctg tgtttccatg attttctaag cgtactctag tgttgttgtt tgtgtctctc gtatatacat t tctct act c ccacaagtta ggtaaatgtt tcctgtcaat cataactgtg tgaaatccct atatcttgac caaacctcct tgctccagtt catccagttg ttacctgcta gactgtagtg aaaagataac tcagaatqac ccttccaagg ttctgcagtt tggtgtcatc tgtaaaggct tgtggttgat tgaattggtt caatgacact tctgtttgca agaccttggt cttcagtgtt taatgaactg tgtatcctca aactgtcgtt taaggctgct gcagatgata gcttaatttt agtcacacta aactacacct ctctccacag catccaagaa ctcaagcagt ggtacctgtg aggctctcag cagcacatct acccagcaat actaaagaaa cactcaagaa tcattccagc ggcctctact tccagtgaca ccaccacagt ggcct cagcc tcctctgcca gcaaggttgg tgcaacatct agtcattcct ccccattatg gatgtatata aaataactat tagttaacca tttaacaatt atgtacagta ctgattgtgt tttaccttta cctttttttg agtgagtctc tgctgtcatg tttcaataaa tttagtagag ctgqttttgg acagatgtca gacacaqttg aCggataagg ttcagattaa tatgagtcca ttgaatcagt ttcacccagt acgaaagtaa ggccctgatg aagaaactag ggqgtaccac aatagcccag catggtacag acgctctcca cgaccaaata gaggatggtg gtcaatgaca ctaccgtcca ggcatgaatg atcgaccaag ttacacagag gtaattgtca gtgcgcaaaa ccaactagtg gtagttattt cagaaaaaca atgatgaaga gtcactattg gaccttccta actactttca cctgccttcc ctgcctctcg tcagatccct tccgtacaca gaaagcagca catggcctgc cgcactgaag gctgcaga aa tgtctcatct tcttcgcaag cagcaccaca cagaccatcg cctcctctag ctctgctaca caggttattg gtgcaaggtg cagttttaca ttgacaacct gaagaacatc gtcaaaattt gtaaatagaa aaaaattgca tgcatcccct ttttttgttg ggaaccagta gtttacctaa ggctttcttt ccttcaaaat ttttctacct gaatatgtat ttcgagacac caagtgatg atgataatgt ttctttcaga atgcggacca ggccagtatt ca aaagaat a cagcaatgct ctttcgtaac gtgcaatgga ctccacctga atagagaaaa ccttaaccag ttttcactca taggactaat ttttagatga tttcatttga gtagagtatc acaaaccagt ctaa tccagg cagaggttcg aaacaggcaa tgttggtcaa atctgttcgt gcactgaagc actatgtcaa tcatcactgc agcagaattc aaaagaaaaa aagaaactaa ttgatctaga agcccgacag aaattcagcc ataacacctt acagcgtttc ccaqaccgtc gtgatggtgg cccttggcta gggatggcaa taactgttca atggccattc cacaggcctc gcccacgagt cattqtgcca tgcaggctac gccctccttt ccctccatcg ctgatgggct ccatgtctga tcactccacg ccttgtaaag aagatacaat acagatacca atttgtttaa tgtacagtaa tttttatcat tgtagcaaat acttttgttc ttgatttttg acgcagtagg tattccaata aaactgtaca agaagtgcaa tggattgatg cccat ccatt aaatattcca taatggcagg cagtaatcag tgccattaaa cttcatcaaa tgtttctatt tgcagacagt attcagcctg agaggataaa caatgtcaca caatgaatac cactgtaact gaatgatgac tagagaaaaa acgttcttca tttcattgtc cacagtggtc ttacagcatt cataacattg agctaatgac gaatgagtcg accagtgacc gatcctggtt tgtagtaaga tgaatgggct gaagaagaag ggcagatgat agagcaaaca ccctgatttg tgaaactccc tgtggcctgt tgactgtggc ccagcggcgt actgggagac tcctcaggag ctccgatcct accaactgtg tgatgcctgc tgctctatgc gacacagacc cagcccacca tgcacttcac agcacaggct tagtcaggcc atgctctgtt aagacttcat ccaacaggcc ctaaaatagt tccaatgagt gaataaatct ttcagaatgt ggcttatcat catgtgcaat ggaaagccta agataacgtt tttgttgttt tagtgtaaat ctatattgtt gatctagatc taactgccct ccagcaagag gacataagat atcaacacca gtgacatgct ttcctcctgg ttactggctg gtgaaagatg cctgagaata gggcctaatg gattgtcgta tatttattca gtctttgtaa aacttctatg gatcctgatt ttcaccattg caagaatctt agtgccaaag cctccttcca tttcagqtaa gtaggaggaa atggagaaat ttaggacagc gtgaccaatg ccaaatactg gcagctgttg tgtcgccagg accccaaacc cattccccta gttgacagtg atgggaaagt gcccgacact ctgaattcga gactctatct tatccagtga gtcacatttc catgatgcag gagtactttg gaatctactt gaagaggcct tggatgccgg cacagcccac attgctctct ccgatacagg cacagcccac gctgcaatca caatcatcag gatcagggag cccagtgatg agaccgtcca tacttcaaat attctgatta acagctagac gtatttaaaa gacagagcgc attactgatt gaaatatctt aaaaggtata tcagtttttt actgcttgtt gataaaattt tacaacctat aattaagcaa 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 00 00 ctatttgtta aaagctgtat aatttgtatg cttaaaatat atttgacagc actgtagata aacttactga tgtaattata cacatgactg tcaaaatctc agaaaacatt attatcttgt catagcaaaa gtttaatggg catagatttt gtaacataaa aaagtttctt atccattgta ttaactatca tgttccaagg tgatatttga tgatgttgaa agtgcttcca gattttttct atgatctaca aaagaagagg gttagtgaaa aatttattga gacacctttc tctgatttta cttggttttt gggtattagc ctggccttgg ttacaagtta ctattgtgat tcaaaaaatg gaatgactta ttccattaga tatgtattca ttacctgtgt tctacaatgt ttttaattat gtagcagtta atgggaaaaa cacacacaca tatttcaaca actccctaat tttatatttt caaatttata cttccaactg tgcaagaacc cttaaaatat tgatttttaa gaaaaatgta tactttttaa tttatgaaat agcatctgaa ttgactattg ttctatttca gaaat aaaagggcct ctgccatttt taacagccta cgacggtaaa t caataggct aaaccatata agaaaaccac aatctacatc cttcttgcag atatatgaaa agttttcttc atgtgcacat ctgagaaaat ggcggttggg gtaatgtata ctgtggtaat ttcatgtgcc aagttcctta gagtttgatt gatttggagc atgtttatta aattttaaac gtagatttta tgtgttttaa cacacacaca ttgaaagatt atcaaatagg gttactaggg ctatagatga taatcagact accagagagg ccagttattc aaacctgtta tttctctgac gtatgatgta gttctgtttt tatgagaata ttgtgattat aatactaagt tttaactaat aataaattta gtaaatttgg caaagagcct aatagactaa aaaaatcctt ttagtttttt acacatttat ttattataat gttagtttag gctcaattgg tttctatatt tctttcacag aagcagttca tgtgcacaaa agagtaaact atgtatttat agtgaacagt tatggccaag ggttctgtat ctttttactt aagcctgtag gaaaattaaa ctactatttt gttaccaaag ctaaatctat taataacaac aaacagaatt tcaacaagaa taaaatatat tgttgtctgt tttataaatg accttgtttc gtgggggggg acttgatatt tgcataattt cagcaggtta agtcatattt aataaaatta atactggttt aattattatc ttggggaaqa gaacattctt ttattttcac cacacacacg attaaataac actcaaagtt aggttcttaa tataggaatg tagattgtgt ctttgaatgg tcccctatgc cttctagggc t gaat agaca tgtagtccta aaaaaggatt atacgttcaa taattttcta atcttatatg attgtgtcag gagagtaatt ttagaaaata ctgccttccc acaaatcaca ttcaatcctg ttgtttgttt gaaaatctac tattatttct gtaaactttt ccgggaaaac atgtgccaat tagctttctt agttgacaac taaaaaaaat gtgtgaaatt gtttgtattg atcccaaagc gatggcagta tgcatgtttt taatagttta tccattatta aggaggtgga gtagagaaac ggtqttcagt aagactaagg aagaatatca ttaaggaaaa ataccaataa tcttctaagc tatttccttc tacagaaaca agtataacac gagtcaatat taatttatga cattggtgag agtagcgttt gactgggcgt attaatgttt tccaggtgca atacaaatgt ttaagaaaag tgcctcaaaa attttctctc tgcacacaca ttatcaggca ggatatttgg atcctcatat aaccaatacg ttagaatatt aagcaggctg cttccttctc ttcagatctg gtggtatagg caactgcaaa ttgtttgatt tcaaagtagt gctatggtat ctacgtgcat tgtattaaaa ttgtgtattc ataataaatg aaactaatat aattgttcag agaaaattaa ctttttcatg ttgtttaggc tatctttctt tattatgacc atgggagcaa taccacacca acactaaccg agcagaaaca taatggcaat tatactatcc tgggaagatt agttccaacc tgtaatccag cttattaata gtgtaaagta cttgggtctt tgcatccaaa tcaggaagat aaaaataaca gatttttgtt ggaaggaact atgcagaggg cacacacaga aaagaaacag ttgtatcctc tcaccaactt taaaccaaga ctcctattga ttaaactgtg gatttccact tcagaatata gcagaataac tttctccttc tgtgaatccc gttgatattg aaccaatagt aacctgcaaa tgcaaaactt cacacattta tctcaatggt gatttttctt ctggaaactt cttttattac aaatgactgg agagtagcca tttctaagcg atgatatctt ttgacacagc acgtcttact tgaaattaaa tattctattt tactatatca acacattctt attagctttt ttatttactt gttagtgcta ttatcacaca ttcttaaaat aggcgtttta gtattactga ttttatttat cttttatatt attagaaact gagaagctga gatcaatttt tcatgtgctt gtaacaaaaa tgtttagtga ctgcttaaaa cctcctctgt atgctttgga aagcaaactt tatattaata catcagaaat tacttctggg gcacgagtca ttaaatgttg aatacatgta attctagctc tttcaagaaa agaaataagg acaaaaacca tactattcat ttaactggcc aattttcttc gacaattgat ttaacttaga tgtaaatttt gaatattgag cattattccc ttcttaactt gtgttgttaa gaaggactga tggctattgt gacaaaaatc gatgatgtga tagttttctg aatgatataa tactatctat ctgacagtat gtgacgtttt cctttctaac gcaccctctt aagaggcaag tccactaggt tttcatcaca acacaagtgg gaaccaacaa acttcaagct tgtgtccata cacttgtgag ttcttaaact acatatgata aacattttac ttgtgtaatg tggtcattaa gtaattatgt ctcacatggc aggtgtgtat actcttctga ttttggaaac attttgaatg aatatatttc atgcagaggc ttagtaaata aatctgctca ttgtaagtga tattaagatt gatatcatac agtaagaagg gtattaattg aaagttatga 5580 5640 5700 5760 5820 5880 5940 6000 6060 6120 6180 6240 6300 6360 6420 6480 6540 6600 6660 6720 6780 6840 6900 6960 7020 7080 7140 7200 7260 7320 7380 7440 7500 7560 7620 7680 7740 7800 7860 7920 7980 8040 8100 8160 8220 8280 8340 8400 8460 8520 8580 8640 8700 8760 8820 8880 8940 9000 9060 9065 268 00Table L11(d). Nucleotide sequence alignment of 109P1D4 v.1 (SEQ ID NO: 258) and 109P1D4 v.5 (SEQ ID NO: 257) Score =7456 bits (3878), Expect =0.Oldentities 387813878 (100%) Strand Plus Plus V.1 1 ctggtggtccagtacctccaaagatatggaatacactcctgaaatatcctgaaaactttt 1 ctggtggtccagtacctccaaagatatggaatacactcctgaaatatcctgaaaactttt V.1 61 ttttttcagaatcctttaataagcagttatgtcaatctgaaagttgcttacttgtacttt 120 61 ttttttcagaatcctttaataagcagttatgtcaatctgaaagttgcttacttgtacttt 120 V.1 121 atattaatagctattcttgtttttcttatccaaagaaaaatcctctaatccccttttcac 180 121 atattaatagctattcttgtttttcttatccaaagaaaaatcctctaatccccttttcac 180 00V.1 181 atgatagttgttaccatgtttaggcattagtcacatcaacccctctcctctcccaaactt 240 00111 11 1111 11111111111I l~~l 181 atgatagttgttaccatgtttaggcattagtcacatcaacccctctcctctcccaaactt 240 V.1 241 ctcttcttcaaatcaaactttattagtccctcctttataatgattccttgcctcgtttta 300 241 ctcttcttcaaatcaaactttattagtccctcctttataatqattccttgcctcgtttta 300 V.1 301 tccagatcaattttttttcactttgatgcccagagctgaagaaatggactactgtataaa 360 301 tccagatcaattttttttcactttgatgcccagagctgaagaaatggactactgtataaa 360 V.1 361 ttattcattgccaagagaataattgcattttaaacccatattataacaaagaataatgat 420 361. ttattcattgccaagagaataattgcattttaaacccatattataacaaagaataatgat 420 V.1 421 tatattttgtgatttgtaacaaataccctttattttcccttaactattgaattaaatatt 480 421 tatattttgtgatttgtaacaaataccctttattttcccttaactattgaattaaatatt 480 V.1 481 ttaattatttgtattctctttaactatcttggtatattaaagtattatcttttatatatt 540 481 ttaattatttgtattctctttaactatcttggtatattaaagtattatcttttatatatt 540 V.1 541 tatcaatggtggacacttttataggtactctgtgtcatttttgatactgtaggtatctta 600 541 tatcaatggtggacacttttataggtactctgtgtcatttttgatactgtaggtatctta 600 V.1 601 tttcatttatctttattcttaatgtacgaattcataatatttgattcagaacaaatttat 660 601 tttcatttatctttattcttaatgtacgaattcataatatttgattcagaacaaatttat 660 V.1 661 cactaattaacagagtgtcaattatgctaacatctcatttactgattttaatttaaaaca 720 661 cactaattaacagagtgtcaattatgctaacatctcatttactgattttaatttaaaaca 720 V.1 :721 gtttttgttaacatgcatgtttagggttggcttcttaataatttcttcttcctcttctct 780 :721 gtttttgttaacatgcatgtttagggttggcttcttaataatttcttcttcctcttctct 780 V.1; 781 ctctcctcttcttttggtcagtgttgtgcgggttaatacaacaaactgtaacaagtgtac 840 269 00 111 ii11111111 1111 iili 11111111 781 ctctcctcttcttttggtcagtgttgtgcgggttaatacaacaaactgtaacaagtgtac 840 V.1 :841 ctggtatggacttgttgtccgggacgtacattttcgcggtcctgctagcatgcgtggtgt 900 :841 ctggtatggacttgttgtccgggacgtacattttcgcggtcctgctagcatgcgtggtgt 900 V.1 :901 tccactctggcgcccaggagaaaaactacaccatccgagaagaaatgccagaaaacgtcc 960 :901 tccactctggcgcccaggagaaaaactacaccatccgagaagaaatgccagaaaacgtcc 960 V.1 :961 tgataggcgacttgttgaaagaccttaacttgtcgctgattccaaacaagtccttgacaa 1020 :961 tgataggcgacttgttgaaagaccttaacttgtcgctgattccaaacaagtccttgacaa 1020 00 V.1 :1021 ctgctatgcagttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaag 1080 :1021 ctgctatgcagttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaag 1080 V. 01agtcggggttccatgcccctgtgggaatttcg14 V.1 1081 aggatactggtgagatcttcactactggcgctcgcattgatcgtgagaaattatgtgctg 1140 V.1: 1141 gtatcccaagggatgagcattgcttttatgaagtggaggttgccattttgccggatgaaa 1200 1141 gtatcccaagggatgagcattgcttttatgaagtggaggttgccattttgccggatgaaa 1200 V.1 :1201 tatttagactggttaagatacgttttctgatagaagatataaatgataatgcaccattgt 1260 1201 tatttagactggttaagatacgttttctgatagaagatataaatgataatgcaccattgt 1260 V.1 :1261 tcccagcaacagttatcaacatatcaattccagagaactcggctataaactctaaatata 1320 :1261 tcccagcaacagttatcaacatatcaattccagagaactcggctataaactctaaatata 1320 V.1 :1321 ctctcccagcggctgttgatcctgacgtaggaatpaacggagttcaaaactacgaactaa 1380 :1321 ctctcccagcggctgttgatcctgacgtaggaataaacggagttcaaaactacgaactaa 1380 V.1: 1381 ttaagagtcaaaacatttttggcctcgatgtcattgaaacaccagaaggagacaagatgc 1440 1381 ttaagagtcaaaacatttttggcctcgatgtcattgaaacaccagaaggagacaagatgc 1440 V.1 :1441 cacaactgattgttcaaaaggagttagatagggaagagaaggatacctacgtgatgaaag 1500 :1441 cacaactgattgttcaaaaggagttagatagggaagagaaggatacctacgtgatgaaag 1500 V.1: 1501 taaaggttgaagatggtggctttcctcaaagatccagtactgctattttgcaagtgagtg 1560 1501 taaaggttgaagatggtggctttcctcaaagatccagtactgctattttgcaagtgagtg 1560 V.1 :1561 ttactgatacaaatgacaaccacccagtctttaaggagacagagattgaagtcagtatac 1620 :1561 ttactgatacaaatgacaaccacccagtctttaaggagacagagattgaagtcagtatac 1620 00 V.1 :1621 cagaaaatgctcctgtaggcacttcagtgacacagctccatgccacagatgctgacatag 1680 :1621 cagaaaatgctcctgtaggcacttcagtgacacagctccatgccacagatgctgacatag 1680 V.1 :1681 gtgaaaatgccaagatccacttctctttcagcaatctagjtctccaacattgccaggagat 1740 :1681 gtgaaaatgccaagatccacttctctttcagcaatctagtctccaacattgccaggagat 1740 V.1: 1741 tatttcacctcaatgccaccactggacttatcacaatcaaagaaccactggatagggaag 1800 1741 tatttcacctcaatgccaccactggacttatcacaatcaaagaaccactggatagggaag 1800 V.1: 1801 aaacaccaaaccacaagttactggttttggcaagtgatggtggattgatgccagcaagag 1860 1801 aaacaccaaaccacaagttactggttttggcaagtgatggtggattgatgccagcaagag 1860 00 V.1 :1861 caatggtgctggtaaatgttacagatgtcaatgataatgtcccatccattgacataagat 1920 :1861 caatggtgctggtaaatgttacagatgtcaatgataatgtcccatccattgacataagat 1920 V.1: 1921 acatcgtcaatcctgtcaatgacacagttgttctttcagaaaatattccactcaacacca 1980 1921 acatcgtcaatcctgtcaatgacacagttgttctttcagaaaatattccactcaacacca 1980 V.1: 1981 aaattgctctcataactgtgacggataaggatgcggaccataatggcagggtgacatgct 2040 1981 aaattgctctcataactgtgacggataaggatgcggaccataatggcagggtgacatgct 2040 V.1 2041 tcacagatcatgaaatccctttcagattaaggccagtattcagtaatcagttcctcctgg 2100 2041 tcacagatcatgaaatccctttcagattaaggccagtattcagtaatcagttcctcctgg 2100 V.1 2101 agactgcagcatatcttgactatgagtccacaaaagaatatgccattaaattactggctg 2160 2101 agactgcagcatatcttgactatgagtccacaaaagaatatgccattaaattactggctg 2160 V.1 :2161 cagatgctggcaaacctcctttgaatcagtcagcaatgctcttcatcaaagtgaaagatg 2220 :2161 cagatgctggcaaacctcctttgaatcagtcagcaatgctcttcatcaaagtgaaagatg 2220 V.1: 2221 aaaatgacaatgctccagttttcacccagtctttcgtaactgtttctattcctgagaata 2280 :2221 aaaatgacaatgctccagttttcacccagtctttcgtaactgtttctattcctgagaata 2280 V.1 :2281 actctcctggcatccagttgacgaaagtaagtgcaatggatgcagacagtgggcctaatg 2340 :2281 actctcctggcatccagttgacgaaagtaagtgcaatggatgcagacagtgggcctaatg 2340 V.1 :2341 ctaagatcaattacctgctaggccctgatgctccacctgaattcagcctggattgtcgta 2400 :2341 ctaagatcaattacctgctaggccctgatgctccacctgaattcagcctggattgtcgta 2400 V.1 :2401 caggcatgctgactgtagtgaagaaactagatagagaaaaagaggataaatatttattca 2460 :2401 caggcatgctgactgtagtgaagaaactagatagagaaaaagaggataaatatttattca 2460 00 V.1 :2461 caattctggcaaaagataacggggtaccacccttaaccagcaatgtcacagtctttgtaa 2520 :2461 caattctggcaaaagataacggggtaccacccttaaccagcaatgtcacagtctttgtaa 2520 V.1 :2521 gcattattgatcagaatgacaatagcccagttttcactcacaatgaatacaacttctatg 2580 :2521 gcattattgatcagaatgacaatagcccagttttcactcacaatgaatacaacttctatg 2580 V.1 :2581 tcccagaaaaccttccaaggcatggtacagtaggactaatcactgtaactgatcctgatt 2640 :2581 tcccagaaaaccttccaaggcatggtacagtaggactaatcactgtaactgatcctgatt 2640 V.1 :2641 atggagacaattctgcagttacgctctccattttagatgagaatgatgacttcaccattg 2700 :2641 atggagacaattctgcagttacgctctccattttagatgagaatgatgacttcaccattg 2700 00 V.1: 2701 attcacaaactggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatctt 2760 2701 attcacaaactggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatctt 2760 V.1: 2761 acactttctatgtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaag 2820 2761 acactttctatgtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaag 2820 V.1 2821 taaccataaatgtggttgatgtcaatgacaacaaaccagttttcattgtccctccttcca 2880 2821 taaccataaatgtggttgatgtcaatgacaacaaaccagttttcattgtccctccttcca 2880 V.1 2881 actgttcttatgaattggttctaccgtccactaatccaggcacagtggtctttcaggtaa 2940 2881 actgttcttatgaattggttctaccgtccactaatccaggcacagtggtctttcaggtaa 2940 V.1 2941 ttgctgttgacaatgacactggcatgaatgcagaggttcgttacagcattgtaggaggaa 3000 2941 ttgctgttgacaatgacactggcatgaatgcagaggttcgttacagcattgtaggaggaa 3000 V.1 3001 acacaagagatctgtttgcaatcgaccaagaaacaggcaacataacattgatggagaaat 3060 3001 acacaagagatctgtttgcaatcgaccaagaaacaggcaacataacattgatggagaaat 3060 V.1 3061 gtgatgttacagaccttggtttacacagagtgttggtcaaagctaatgacttaggacagc 3120 3061 gtgatgttacagaccttggtttacacagagtgttggtcaaagctaatgacttaggacagc 3120 V.1 3121 ctgattctctcttcagtgttgtaattgtcaatctgttcgtgaatgagtcggtgaccaatg 3180 3121 ctgattctctcttcagtgttgtaattgtcaatctgttcgtgaatgagtcggtgaccaatg 3180 V.1: 3181 ctacactgattaatgaactggtgcgcaaaagcactgaagcaccagtgaccccaaatactg 3240 3181 ctacactgattaatgaactggtgcgcaaaagcactgaagcaccagtgaccccaaatactg 3240 V.1 :3241 agatagctgatgtatcctcaccaactagtgactatgtcaagatcctggttgcagctgttg 3300 :3241 agatagctgatgtatcctcaccaactagtgactatgtcaagatcctggttgcagctgttg 3300 00 V.1: 3301 ctggcaccataactgtcgttgtagttattttcatcactgctgtagtaagatgtcgccagg 3360 3301 ctggcaccataactgtcgttgtagttattttcatcactgctgtagtaagatgtcgccagg 3360 V.1 :3361 caccacaccttaaggctgctcagaaaaacaagcagaattctgaatgggctaccccaaacc 3420 3361 caccacaccttaaggctgctcagaaaaacaagcagaattctgaatgggctaccccaaacc 3420 V.1 3421 cagaaaacaggcagatgataatgatgaagaaaaagaaaaagaagaagaagcattccccta 3480 3421 cagaaaacaggcagatgataatgatgaagaaaaagaaaaagaagaagaagcattccccta 3480 V.1 3481 agaacttgctgcttaattttgtcactattgaagaaactaaggcagatgatgttgacagtg 3540 3481 agaacttgctgcttaattttgtcactattgaagaaactaaggcagatgatgttgacagtg 3540 V.1 3541 atggaaacagagtcacactagaccttcctattgatctagaagagcaaacaatgggaaagt 3600 3541 atggaaacagagtcacactagaccttcctattgatctagaagagcaaacaatgggaaagt 3600 V.1 3601 acaattgggtaactacacctactactttcaagcccgacagccctgatttggcccgacact 3660 3601 acaattgggtaactacacctactactttcaagcccgacagccctgatttggcccgacact 3660 V.1 3661 acaaatctgcctctccacagcctgccttccaaattcagcctgaaactcccctgaattcga 3720 V. 5 3661 acaaatctgcctctccacagcctgccttccaaattcagcctgaaactcccctgaattcga 3720 V. 1 3721 agcaccacatcatccaagaactgcctctcgataacacctttgtggcctgtgactctatct 3780 V. 5 3721 agcaccacatcatccaagaactgcctctcgataacacctttgtggcctgtgactctatct 3780 V.1 3781 ccaagtgttcctcaagcagttcagatccctacagcgtttctgactgtggctatccagtga 3840 3781 ccaagtgttcctcaagcagttcagatccctacagcgtttctgactgtggctatccagtga 3840 V.1 3841 cgaccttcgaggtacctgtgtccgtacacaccagaccg 3878 3841 cgaccttcgaggtacctgtgtccgtacacaccagaccg 3878 Table UV(d). Peptide sequences of protein coded by 109PiD4 v.5 (SEQ ID NO: 258) MDLLSGTYIF AVLLACVVFH SGAQEKNYTI REEMPENVLI GDLLKDLNLS MQFKLVYKTG DVPLIRIEED TGEIFTTGAR IDREKLCAGI PRDEHCFYEV RLVKIRFLIE DINDNAPLFP ATVINISIPE NSAINSKYTL PAAVDPDVGI SQNIFGLDVI ETPEGDKMPQ LIVQKELDRE EKDTYVMT{VK VEDGGFPQRS DTNDNHPVFK ETEIEVSIPE NAPVGTSVTQ LHATDADIGE NAKIHFSFSN HLNATTGLIT IKEPLDREET PNHKLLVLAS DGGLMPARAM VLVNVTDVND VNPVNDTVVL SENIPLNTKI ALITVTDKDA DHNGRVTCFT DHEIPFRLRP AAYLDYESTK EYAIKLLAAD AGKPPLNQSA MLFIKVKDEN DNAPVFTQSF PGIQLTKVSA MDADSGPNAK INYLLGPDAP PEFSLDCRTG MLTVVKKLDR LAKDNGVPPIJ TSNVTVFVSI IDQNDNSPVF THNEYNFYVP ENLPRHGTVG DNSAVTLSIL DENDDFTIDS QTGVIRPNIS FDREKQESYT FYVKAEDGGR INVVDVNDNK PVFIVPPSNC SYELVLPSTN PGTVVFQVIA VDNDTGMNAE RDLFAIDQET GNITLMEKCD VTDLGLHRVL VKANDLGQPD SLFSVVIVNL LINELVRKST EAPVTPNTEI ADVSSPTSDY VKILVAAVAG TITVVVVIFI LI PNKSLTTA

EVAILPDEIF

NGVQNYELIK

STAILQVSVT

LVSNIARRLF

NVPSIDIRYI

VFSNQFLLET

VTVS IPENNS

EKEDKYLFTI

LI TVTDPDYG

VSRSSSAKVT

VRYS IVGGNT

FVNESVTNAT

TAVVRCRQAP

00 oHLKAAQKNKQ NqSEWATPNPE NRQMIMMKKK KKKKKHSPKN LLLNPVTIEE TKADDVDSDG 900 ONRVTLDLPID LEEQTMGKYN WVTTFTTFKP DSPDLARHYK SASPQPAFQI QFETFLNSKH 960 ClHIIQELPLDN TFVACDSISK CSSSSSDPYS VSDCGYPVTT FEVPVSVHTR PSQRRVTFHL 1020 PEGSQESSSD GGLGDHDAGS LTSTSHGLPL GYPQEEYFJR ATPSNRTEGD GNSDPESTFI 1080 PGLKKAAEIT VQPTVEEASD NCTQECLIYG HSDACWM~PAS LDHSSSSQAQ ASALCHSPPL 1140 COSQASTQHHSP RVTQTIALGH SPFVTQTIAL CHSEPPIQVS ALHHSPPLVQ ATALHHSPPS 1200 AQASAIJCYSP PLAQAAAISII SSPLPQVIMJ HRSQAQSSVS LQQGWVQ-GAD GLCSVDQGVQ 1260 GSATSQFYTM SERLHPSDDS IKVIPLTTFT PRQQARPSRG DSPIMEEHPL 1310 Table LV(d). Amino acid sequence alignment of IP10 4v.1 (SEQ ID NO: 259) and 109PI1D4 v.5 (SEQ ID NO: 260) 0 Score =2005 bits (5195), Expec O.Oldentities 10111/1011 Posiflves 1011 /1011 (100%) V. DLGYCVLCVFSAlNTREPNLGDLDNSINSTA6 O v.1 1 MDLLSGTYIFAVLLACVVFHSGAQEKNYTIREEMPENVLIGDLLKDLWLSLIPNCSLTTA 6 1 HDLLSGTYIFAVLLACVVFHSGAQEKMYTIREEMPENVLIGDLLKDLNLSLIPNKSLTTA 6 00 V.1 61 MQFKLVYKTGDVPLIRIEEDTGEIPTTGARIDRE1CLCAGIPROEHCFYEVEVAILPDEIF 120 O MQFKLVYKTGDVPLIRIEEDTGEI ETTGARIDREKLCAGIPRD)EHCFYEVEVAILPDEIF 61 MQFKLVYKTGDVPLIRIEEDTGEIrI!TGARIOREKLCAGIPRDEHCE'YEVEVAILPDEIF 120 V. 2 CKRLEIDALPTIIIPNANKTPADDGNVNlI 8 V.1 121RLVKIRFLIEDINDNAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVQNYELIK18 121RLVKIRFLIEDINDNAPLFPATVINIS IPENSAINSKYTLPAAVDPDVGINGVQNYELIC18 V.1 121 RLVIFLIEINDAPLAVINISIPEAKYTLPAVDGPVGINGVQYELI 180 V.1 181SQNIFGLDVIETPEGDO4PQLIVQKELDREEKDTYVNKVKVEDGGFPQRSSTAILQVSVT24 181SQNI FGLDVIETPEGDKMPQLIVQKELDREEKDTYVNICVKVEDGGFPQRSSTAILQVSVT24 2411 SQNIFGLDVIETIIEGD NLI VQDREE DV VKDGGFQSLSAQVS 2400 V.1 241DTNDNHPVFKETEIEVSIPENAPVGTSVTQLHATDADIGENAKIHFSFSNLVSNIARRLF30 241DTNDNHPVFKETEIEVSIPENAPVGTSVTQL.HATDADIGENAKIHFSFSNLVSNIARRLF30 V.1 241 DLTDNHPVKEEIEPNVGTVTLADMADIRAXIHFSFSDNLVSIRL V.1 301HLNATTGLITIKEPLDREETPNHKLLVJASDGGLMPARAMVLVNVTDVNDNVPSIDIRYI36 301HLNATTGLITIKEPLDREETPNHKLLVLASDGGLMPARAMVLVNVTDVNDNVPSIDIRYI36 301 HLPNTGLIPLRETPKLVLKADGGMPPAMVLVNVTDRVDNPSIDIRY 360 V.1 361VNPVND'PVVLSE1IPLNTKIALITVTDKDADHNGRVTCFTDHEIEFRLRPVFSNQFLLET42 361VNPVNDTVVLSENIPLNTKIALITVTDKDADHNGRVTCFTDHEIPFRLRPVFSNQFLLET42 V.1 361 VNPVDTVVLENAIPLTDKIALDKADHNGRVTCETDHEPFPVF TSQFLET 420 V.1 421AAYLDYESTKEYAIflJLAADAGKPPLNQSAI4LFIKVKDENDNAPVFI'QSFVTVS IPENNS48 421AAYLDYESTKEYAIKLLAADAGKPPLNQSAMLFIKVKDENDNAPVFTQSFVTVS IPENNS48 421 AAYQLDTKEAIKLLAADNAKNGPPSMLKDD NAPLVETVV SPENNSFT 540 V.1 481PGIQLTKVSAMDADSGPNAKINYLLCPDAPPEFSLDCRTGMLTVVKKLDREKEDKYLETI54 481PGIQLTKVSAMDADSGPNAKINYLLGPDAPPEFSLDCRTGMLTVVKKLDREKEDKYLFTI54 5481 PLKVPSADADSGPAIDNLDPFHEPESRGLVVKKLEK DY 500 V.1 541LAKDNGVPPLTSWVTVFVSIIDQNDNSPVF7HNEYNFYVPENLPRHGTVGLITVTDPDYG60 541LAKDNGVPPLTSNVTVFVSIIDQNDNSPVETHNEYNFYVPENLPRHGTVGLITVTDPDYG60 5461 LDNVLTSNVTVFVIIDQNDRNSENEF YNLPRGVLITVTDPDY V.1 601DNSAVTLSILDENDDF'TIDSQTGVIRPNISFDREKESYTFYVKAEDGGRVSRSSSAKVT60 601DNSAVTLSILDENDDFTIDSQTGVIRPNISFDREKQESYTFYVKAEDGGRVSRSSSAKVT60 601 DNSADVTLIDEWDFIOSSTGVRPSNISFDREKQYVAD GGRAVSRYSSSAKT 660 V.1 661INVVDVNDNKPVFIVPPSNCSYfELVLPSTNPGTVVFQVIAVDNTGMNAEVRYSIVGGNT72 661INVVDVNDNKPVFIVPPSNCSYELVLPSTNPGTVVFQVIAVDNDTGM4NAEVRYSIVGGNT72 661 INVVOVDNKPVIVPPSNCSYEDLGVVLP1PGTWQIVDTGMINEVYSVGNAT 720 V.1 721RDLFAIDQETGNITLMKCDVTDLGLHRVLVKA24DLGOPDSLFSVVIVNLFVNESVTNAT78 721RDLFAIDQETG4ITLHEKCDVTDLGLHRVLVKANDLGOPDSLFSVVIVNLFVNESVTNAT78 721 RLIDQVRSEGNI TECDVDLGLHRVVKIADLGPDSLFVVINFVNEVTNRAT 840 V.1 781LINELVEKSTEAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVVVIFITAVVRCRQAP84 781LINELVRKSTEAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVVVIFITAVVRCRQAP84 00 00 V.1 841 HLKAAQKNKQNSEWATPNPENRQMIMIKKKKKKHSPKNLLLNFVTIEETKADDVDSDG 900

HLKAAQKNKQNSEWATPNPENRQMIMMKKKKKICHSPKNMLLLNEVTIEETKADDVDSDG

841 HLKAAQKNKQNSEWATPNPENRQMIMKKKKKKKHSPKNLLLNFVTIEETKADDVDSDG 900 V.1 901 NRVTLDLPIDLEEQTMGKYNVTTPTTFKPDSPDLARHIYKSASPQPAFQIQPETPLNSKH 960

NRVTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLARHYKSASPQPAFQIQPETPLNSKH

901 NRVTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLIARHYKSASPQPAFQIQPETPLNSKH 960 V.1 961 HIIQELPLDNTFVACDSISKCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP 1011

HIIQELPLDNTFVACDSISKCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP

961 HIIQELPLDNTFVACDSISKCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP 1011 Table UI(e). Nucleotide sequence of transcript variant 109P1 D4 v.6 (SEQ ID NO: 261) ggcagtcggc gcgcggcaaa acaggcagca ggctgctctg tggagcgggc atcgttatta ggatggaata aatacagaat gaaaatcctg agctgctctc ctatgaqgac tacaacaaac atgcgtggtg agaaaacgtc gtccttgaca tcgaattgaa attatgtgct gccggatgaa tgcaccattg ctctaaatat ctacgaacta agacaagatg tgtgatgaaa gcaagtaagt agtcagtata tgctgacata tgccaggaga ggatagggaa gccagcaaga tgacataaga actcaacacc ggtgacatgc gttcctcctg attactggct agtgaaagat tcctgagaat tgggcctaat ggatcgtcgt atatttattc agtctttgta caaattctat tgatcctgat cttcaccatt acaagaatct aagtgccaaa ccctccttac ctttcaggta tgtaggagga gatggagaaa gaactgtctg gaggaaggca caggcagccc cggactgcga ggaagccttt gaqggtgctt tcttaacaaa gtgaagtatc ttgtgaataa aactttgtaa tgaatgacag tgtcacaagt ttccactctg ctgataggca actactatgc gaggatactg ggtatcccaa atatttagac ttcccagcaa actctcccag attaagagtc ccacaactga gtaaaggttq gttactgata ccagaaaatg ggtgaaaatg ttatttcacc gaaacaccaa gcaatggtgc tacatcgtca aaaattgctc ttcacagatc gagaatgcag gcagatgctg gaaaatgaca aactctcctg gctgagatca acaggcatgc acaattctgg agcattattg gtcccagaaa tatggagaca gattcacaaa tacactttct gtaaccataa aactattctt attgctgttg aacacaagag tgtgatgtta ggcgggagga agccaaacag gggctgcctg gctgtggcgg tttctccctt aaaaagtaca acacattttc tctgaactgt gaaggattcc tcttgtgaag tgggttttaa gtttgttgtc gcgcccagga acttgttgaa agttcaagct gtgagatctt gggatgagca tggttaagat cagttatcaa cggctgttga aaaacatttt ttgttcaaaa aagatggtgg caaatgacaa ctcctgtagg ccaagatcca tcaatgccac accacaagtt tggtaaatgt atcctgtcaa tcataactgt atgaaattcc catatcttga gcaaacctcc atgctccagt gcatccagtt attacctgct tgactgtagt caaaagataa atcagaatga accttccaag attctgcagt ctggtgtcat atgtaaaggc atgtggttga atgaattggt acaatgacac atctgtttgc cagaccttgg gccgtgaqca agtgcgcaga aatagcctca tagagcccgc tcgtttacct gatcaactgg cttaagtaaa gctgttgaat acagatcaca aagctgacaa ttcagatatt cgggacgtac gaaaaactac agaccttaac agtgtacaag cactaccggc ttgcttttat acgttttctg catatcaatt tcctgacgta tggcctcgat ggagttagat ctttcctcaa ccacccagtc cacttcagtg CttCtCtttC cactggactt actggttttg tacagatgtc tgacacagtt gacggataag tttcagatta ctatgagtcc tttgaatcag tttcacccag gatgaaagta aggccctgat gaagaaacta tggggtacca caatagccca gcatggtaca tacgctctcc ccgaccaaat tgaggatggt tgtcaatgac tctaccgtcc tggcatgaat aatcgaccaa tttacacaga gtagctgcac gtggcagtgc gaaacaacct tacagcagtc cttcattcta atggatgaat ttcatgcata atggtagcta taccagagcg gcttggctga tcaagtgttg attttcgcgg accatccgag ttgtcgctga accggagatg gctcgcattg gaagtggagg atagaagata ccaqaqaact ggcataaacg gtcattgaaa agggaagaga agatccagta tttaaggaga acacagctcc agcaatctag atcacaatca gcaagtgatg aatgataatg gttctttcag gatgcggacc aggccagtat acaaaagaat tcagcaatgc tctttcgtaa agtgcaacgg gctccacctg gatagagaaa cccttaacca gttttcactc gtaggactaa attttagatg atttcatttg ggtagagtat aacaaaccag actaatccag gcagaggtt c gaaacaggca gtgttggtca tcagctgccc cagcggcgac cagcgactcc gcagtct ccg ctctaaaggc ggatggaaga ctccaaataa ctagctacat gttttgcctc ttgcagtgca tgcgggttaa tcctgctagt aagaaattcc ttccaaacaa tgccactgat atcgtgagaa ttgccatttt taaatgataa cgqctataaa gagttcaaaa caccagaagg aggataccta ctgctatttt cagagattga atgccacaga tctccaacat aagaaccact qtggattgat tcccatccat aaaatattcc ataatggcag tcagtaatca atgccattaa tcttcatcaa ctgtttctat atgcagacag aattcagcct aagaggataa gcaatgtcac acaatgaata tcactgtaac agaatgatga atagagaaaa cacgttcttc ttttcattgt gcacagtggt gttacagcat acataacatt aagctaatga 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 00 00 cttaggacag agtgaccaat cccaaatact tgcagctgtt atgtcgccag taccccaaac gcattcccct tgttgacagt aatgggaaag ggcccgacac cctgaatttg tgactctatc ctatccagtg gacatgaact ccccttccaa gtaatctaga gaaccttgtg cagtgcagca ccatgatgta aacatgcaat ttggaagtga tttaagaccc tattcactaa gttgagtggg cgggtggatc tccactaaaa tcgggaggct gatggcgcca aaaatgttca ttccactcct aatcccagca agtntggcca cctgattctc gctacactga gagatagctg gctggcacca gcaccacacc ccagaaaaca aagaacctgc gatggaaaca tacaattggg tacaaatctg aagcaccaca tccaagtgtt acaaccttcg attgaaatct aaaatttcaa tttcccatta ctttctttag caataacaga atataaggct tacttgccct gctgaactag caaacaaaca cataatattg ccgggCgCgg acgaggtcag atacaaaaaa gaggcaggag ctctgcactc atgatagaaa tttaattatt ctttgggagg acatggcgaa tcttcagtgt ttaatgaact atgtatcctc taactgtcgt ttaagqctgc ggcagatgat tgcttaattt gagtcacact taactacacc cctctccaca tcatccaaga cctcaagcag aggtacctgt gcagtgagat tggattgtga taaaagcaag ctgtaatctg gtactctcat gtcttggtgt gtctgattgt ccaaactact aaaaacaaaa c tgagaaaat tggctcacgc gagattgaga ttagcctggc aatagcgtga cagcctgggt ataattttac aaaaagttat ccgaggtggg accccgtttt tgtaattgtc ggtgcgcaaa accaactagt tgtagttatt tcagaaaaac aatgatgaag tgtcactatt agaccttcct tactactttc gcctgccttc actgcctctc ttcagatccc gtccgtacac gtaactttct tttcaaaatt aatctgttcg agcattgaag gactatgtca ttcatcactg atgcagaatt aaaaagaaaa gaagaaacta attgatctag aagcctgaca caaattcagc gataacacct tacagcgttt accagaccga aggaacaaca aggctaagat caaaaatcat cttaaaaatg tgaatgagtc caccaqtqac agatcctggt ctgtagtaag ctgaatgggc agaagaagaa aggcagatga aagagcaaac gccctgattt ctgaaactcc ttgtgg(Cctg ctgactgtgg ctgattccag aaattccatt cattaatttt atgtcctagt atggaagaga gaatcaacag tcagtcatga ttccaggagt aagagacatt gttttgaaaa ctgcttaaaa gaggccgagg aaaccccatc tcccagctac agtgagccga caaaaagaaa actcatggtg ctcacaccgt gttcaagacc 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4830 gcaatggaaa gctgtttctc atacacttat tgaataatta ctctgaaagg ccaaaacact catttttatt ctgtaatccc ccatcctggc qtggtggcgg acccgggagg gacagagcaa taggttttta ttttggggtg tggatcacct tttaaaattt tgtttgctct ggttaatata aaacattatc tatccagggc ctggttcagt acccaccact agcactttgg taacacggtg gcgcctgtag cggagcttgc gactctqtct tgttgattgt ggtgtggtgg gaggtcagga Table Ull(e). Nucleotide sequence alignment of 109P1D4 v.1 (SEQ ID NO: 262) and 109P104 v.6 (SEQ ID NO: 263) Score =5676 bits (2952), Expect 0.Oldenfties =3002/3027 Strand =Plus Pius V.1 852 ttgttgtccgggacgtacattttcgcggtcctgctagcatgcgtggtgttccactctggc 911 V.6 683 ttgttgtccgggacgtacattttcgcggtcctgctagtatgcgtggtgttccactctggc 742 V.1 912 gcccaggagaaaaactacaccatccgagaagaaatgccagaaaacgtcctgataggcgac 971 V.6 743 gcccaggagaaaaactacaccatccgagaagaaattccagaaaacgtcctgataggcaac 802 V.1 972 ttgttgaaagaccttaacttgtcgctgattccaaacaagtccttgacaactgctatgcag 1031 V.6 803 ttgttgaaagaccttaacttgtcgctgattccaaacaagtccttgacaactactatgcag 862 V.1 1032 ttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaagaggatactggt 1091 VA6 863 ttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaagaggatactggt 922 V. 1 1092 gagatcttcactactggcgctcgcattgatcgtgagaaattatgtgctggtatcccaagg 1151 V.6 923 gagatcttcactaccggcgctcgcattgatcgtgagaaattatgtgctggtatcccaagg 982 V.1 1152 gatgagcattgcttttatgaagtggaggttqccattttgccggatgaaatatttagactg 1211 V. 6 983 gatgagcattgcttttatgaagtggaggttgccattttgccggatgaaatatttagactg 1042 00 V.1: 1212 gttaagatacgttttctgatagaagatataaatgataatgcaccattgttcccagcaaca 1271 V.6: 1043 gttaagatacgttttctgatagaagatataaatgataatgcaccattgttcccagcaaca 1102 V.1 :1272 gttatcaacatatcaattccagagaactcggctataaactctaaatatactctcccagcg 1331 V.6 :1103 gttatcaacatatcaattccagagaactcggctataaactctaaatatactctcccagcg 1162 V.1 :1332 gctgttgatcctgacgtaggaataaacggagttcaaaactacgaactaattaagagtcaa 1391 V.6 :1163 gctgttgatcctgacgtaggcataaacggagttcaaaactacgaactaattaagagtcaa 1222 V. 32actttgctgttataaccaagaaagtgccatat15 00V.1 123 aacatttttggcctcgatgtcattgaaacaccagaaggagacaagatgccacaactgatt 1451 C1V.1 :1452 gttcaaaaggagttagatagggaagagaaggatacctacgtgatgaaagtaaaggttgaa 1511 V. 6 :1283 gttcaaaaggagttagatagggaagagaaggatacctatgtgatgaaagtaaaggttgaa 1342 V.1 :1512 gatggtggctttcctcaaagatccagtactgctattttgcaagtgagtgttactgataca 1571 V.6 1343 gatggtggctttcctcaaagatccagtactgctattttgcaagtaagtgttactgataca 1402 V.1 1572 aatgacaaccacccagtctttaaggagacagagattgaagtcagtataccagaaaatgct 1631 V.6 1403 aatgacaaccacccagtctttaaggagacagagattgaagtcagtataccagaaaatgct 1462 V.1 1632 cctgtaggcacttcagtgacacagctccatgccacagatgctgacataggtgaaaatgcc 1691 V. 6 1463 cctgtaggcacttcagtgacacagctccatgccacagatgctgacataggtgaaaatgcc 1522 V.1 1692 aagatccacttctctttcagcaatctagtctccaacattgccaggagattatttcacctc 1751 V. 6 1523 aagatccacttctctttcagcaatctagtctccaacattgccaggagattatttcacctc 1582 V. 1 1752 aatgccaccactggacttatcacaatcaaagaaccactggataggaagaaacaccaaac 1811 V.6 :1583 aatgccaccactggacttatcacaatcaaagaaccactggatagggaagaaacaccaaac 1642 V.1: 1812 cacaagttactggttttggcaagtgatggtggattgatgccagcaagagcaatggtgctg 1871 V.6 1643 cacaagttactggttttggcaagtgatggtggattgatgccagcaagagcaatggtgctg 1702 V.1 1872 gtaaatgttacagatgtcaatgataatgtcccatccattgacataagatacatcgtcaat 1931 V.6 1703 gtaaatgttacagatgtcaatgataatgtcccatccattgacataagatacatcgtcaat 1762 V.1 1932 cctgtcaatgacacagttgttctttcagaaaatattccactcaacaccaaaattgctctc 1991 V.6 1763 cctgtcaatgacacagttgttctttcagaaaatattccactcaacaccaaaattgctctc 1822 V.1 1992 ataactgtgacggataaggatgcggaccataatggcagggtgacatgcttcacagatcat 2051 277 00v.6 1823 ataactgtgacggataaggatgcggaccataatggcagggtgacatgcttcacagatcat 1882 V.1 2052 gaaatccctttcagattaaggccagtattcagtaatcagttcctcctggagactgcagca 2111 V.6: 1883 gaaattcctttcagattaaggccagtattcataatcagttcctcctggagaatgagca 1942 V.1: 2112 tatcttgactatgagtccacaaaagaatatgccattaaattactggctgcagatgctggc 2171 V.6 :1943 tatcttgactatgagtccacaaaagaatatgccattaaattactggctgcagatgctggc 2002 cIV.1 :2172 aaacctcctttgaatcagtcagcaatgctcttcatcaaagtgaaagatgaaaatgacaat 2231 V.6: 2003 aaacctcctttgaatcagtcagcaatgctcttcatcaaagtgaaagatgaaaatgacaat 2062 00V.1: 2232 gctccagttttcacccagtctttcgtaactgtttctattcctgagaataactctcctggc 2291 00lli iiIlI~ll1II1l~ ii1111111111 V.6: 2063 gctccagttttcacccagtctttcgtaactgtttctattcctgagaataactctcctggc 2122 V. 22acatgagagaggatgagaaatgctatcagtat25 V.1 2122 atccagttgacgaaagtaagtgcaatggatgcagacagtgggcctaatgctaagatcaat 231 V.6 212 acctag ctgatgatcaacctatgcggatgcaggacagt ggcct atgctgat 2412 V.61 213 tacctgctaggccctgatgctccacctgaattcagcctggatgtcgtacaggcatgctg 2241 V.1 2412 actgtagtgaagaaactagatagagaaaaagaggataaatatttattcacaattctggca 2471 V.6 2243 actgtagtgaagaaactagatagagaaaaagaggataaatatttattcacaattctggCa 2302 V.1 2472 aaagataacggggtaccacccttaaccagcaatgtcacagtctttgtaagcattattgat 2531 V. 6 2303 aaagataatggggtaccacccttaaccagcaatgtcacagtctttgtaagcattattgat 2362 V. 1 2532 cagaatgacaatagcccagttttcactcacaatgaatacaacttctatgtcccagaaaac 2591 V.6 2363 cagaatgacaatagcccagttttcactcacaatgaatacaaattctatgtcccagaaaac 2422 V.1 2592 cttccaaggcatggtacagtaggactaatcactgtaactgatcctgattatggagacaat 2651 V.6: 2423 cttccaaggcatggtacagtaggactaatcactgtaactgatcctgattatggagacaat 2482 V.1: 2652 tctgcagttacgctctccattttagatgagaatgatgacttcaccattgattcacaaact 2711 V.6 2483 tctgcagttacgctctccattttagatgagaatgatgacttcaccattgattcacaaact 2542 V.1 :2712 ggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatcttacactttctat 2771 V.6: 2543 ggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatcttacactttctat 2602 V.1 :2772 gtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaagtaaccataaat 2831 VA 2603 gtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaagtaaccataaat 2662 V.1 :2832 gtggttgatgtcaatgacaacaaaccagttttcattgt~cctccttccaactgttcttat 2891 278 00 V-6 2663 gtggttgatgtcaatgacaacaaaccagttttcattgtccctccttacaactattcttat 2722 V. 82gatgtcactccatcggaatgNtcgtatgttgc25 V.6 2823 gaattggttctaccgtccactaatccaggcacagtggtctttcaggtaattgctgttgac 2951 V.1 2952 aatgacactggcatgaatgcagaggttcgttacagcattgtaggaggaaacacaagagat 3011 V.6 2783 aatgacactggcatgaatgcagaggttcgttacagcattgtaggaggaaacacaagagat 2842 V.1 3012 ctgtttgcaatcgaccaagaaacaggcaacataacattgatggagaaatgtgatgttaca 3071 V.6 283ctgtttgcaatcgaccaagaaacaggcaacataacattgatggagaaatgtgatgttaca 2902 00 V.1 3072 gaccttggtttacacagagtgttggtcaaagctaatgacttaggacagcctgattctctc 3131 V.6 :2903 gaccttggtttacacagagtgttggtcaaagctaatgacttaggacagcctgattctctc 2962 V. 12tcggtgatgcacgtgtatatgtacatcaatat39 V.61 3132 ttcagtgttgtaattgtcaatctgttcgtgaatgagtcggtgaccaatgctacactgatt 3191 V.1 :3192 aatgaactggtgcgcaaaagcactgaagcaccagtgaccccaaatactgagatagctgat 3251 V.6 :3023 aatgaactggtgcgcaaaagcattgaagcaccagtgaccccaaatactgagatagctgat 3082 V.1 :3252 gtatcctcaccaactagtgactatgtcaagatcctggttgcagctgttgctggcaccata 3311 V. 6 :3083 gtatcctcaccaactagtgactatgtcaagatcctggttgcagctgttgctggcaccata 3142 V.1: 3312 actgtcgttgtagttattttcatcactgctgtagtaagatgtcgccaggcaccacacctt 3371 V.6: 3143 actgtcgttgtagttattttcatcactgctgtagtaagatgtcgccaggcaccacacctt 3202 V.1 3372 aaggctgctcagaaaaacaagcagaattctgaatgggctaccccaaacccagaaaacagg 3431 V.6 3203 aaggctgctcagaaaaacatgcagaattctgaatgggctaccccaaacccagaaaacagg 3262 V.1 3432 cagatgataatgatgaagaaaaagaaaaagaagaagaagcattcccctaagaacttgctg 3491 lii! Ii 1111IIIIIIIIIIIII 111111111111111 Hill11 V.6 :3263 cagatgataatgatgaagaaaaagaaaaagaagaagaagcattcccctaagaacctgctg 3322 V.1: 3492 cttaattttgtcactattgaagaaactaaggcagatgatgttgacagtgatggaaacaga 3551 V.6: 3323 cttaattttgtcactattgaagaaactaggcagatgatgttgacagtgatggaaacaga 3382 V.1 3552 gtcacactagaccttcctattgatctagaagagcaaacaatgggaaagtacaattgggta 3611 V.6: 3383 gtcacactagaccttcctattgatctagaagagcaaacaatgggaaagtacaattgggta 3442 V.1: 3612 actacacctactactttcaagcccgacagccctgatttggcccgacactacaaatctgcc 3671 V.6 :3443 actacacctactactttcaagcctgacagccctgatttggcccgacactacaaatctgcc 3502 V. 1 :3 672 tctccacagcctgccttccaaattcagcctgaaactcccctgaattcgaagcaccacatc 3731 V.6 :3503 tctccacagcctgccttccaaattcagcctgaaactcccctgaatttgaagcaccacatc 3562 V.1 ;3732 atccaagaactgcctctcgataacacctttgtggcctgtgactctatctccaagtgttcc 3791 V.6 :3563 atccaagaactgcctctcgataacactttgtggcctgtgactctatctccaagtgttcc 3622 V.1 :3792 tcaagcagttcagatccctacagcgtttctgactgtgqctatccagtgacgaccttcgag 3851 V. 6 :3623 tcaagcagttcagatccctacagcgtttctgactgtggctatccagtgacaaccttcgag 3682 V. 1 :3852 gtacctgtgtccgtacacaccagaccg 3878 V.6 :3683 gtacctgtgtccgtacacaccagaccg 3709 Table LIV(e). Peptide sequences of protein coded by 109PI D4 v.6 (SEQ ID NO: 264) MTVGFNSDIS SVVRVNTTNC HKCLLSGTYI FAVLLVCVVF HSGAQEKNYT IGNLLKDLNL SLIPNKSLTT TMQFKLVYKT GDVPLIRIEE DTGEIFTTGA IPRDEHCFYE VEVAILPDEI FRLVKIRFLI EDINDNAPLF PATVINISIP LPAAVDPDVG INGVQNYELI KSQNIFGIJDV IETPEGDKMP QLIVQKELDR KVEDGGFPQR SSTAILQVSV TDTNDNHPVF KETEIEVSIP ENAPVGTSVT ENAKIHFSFS NLVSNIARRL FHLNATTGLI TIKEPLDREE TPNHKIJLVLA MVILVNVTDVN DNVPSIDIRY IVNPVNDTVV LSENIPLNTK IALITVTDKD TDHEIPFPJLR PVFSNQFLLE NAAYLDYEST KEYAIKLLAA DAGKPPLNQS NDNAPVFTQS FVTVSIPENN SPGIQLMKVS ATDADSGPNA EINYLLGPDA GMLTVVKKIJD REKEDKYLFT ILAKDNGVPP LTSNVTVFVS IIDQNDNSPV PENLPRHGTV GLITVTDPDY GDNSAVTLSI LDENDDFTID SQTGVIRPNI TFYVKAEDGG RVSRSSSAKV TINVVDVNDN KPVFIVPPYN YSYELVLPST AVDNDTGMNA EVRYSIVGGN TRDLFAIDQE TGNITLMEKC DVTDLGLHRV DSLFSVVIVN LFVNESVTNA TLINEIJVRKS IF.APVTPNTE IADVSSPTSD GTITVVVVIF ITAVVRCRQA PHLKAAQKNM QNSEWATPNP ENRQMIMMKK NLLLNFVTIE ETKADDVDSD GNRVTLDLPI DLEEQTMGKY NWTTPTTFK KSASPQPAFQ IQPETPLNLK HHIIQELPLD NTFVACDSIS KCSSSSSDPY TFEVPVSVHT RPTDSRT IREEI PENVL RI DREKLCAG

ENSAINSKYT

EEKDTYVMKV

QLHATDADIG

SDGGLMPARA

ADHNGRVTCF

AMLFIKVKDE

PPEFSLIJRRT

FI'HNEYKFYV

SFDREKQESY

NPGTVVFQVI

LVKANDLGQP

YVKIILVAAVA

KKKKKKHS PK

PDSPDLARHY

SVS DCGYPVT 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1037 Table LV(e). Amino acid sequence alignment of 109P1 D4 v.1 (SEQ ID NO: 265) and 109P1 D4 v.6 (SEQ ID NO: 266) Score =1966 bits (5093), Expect 0.Oldentities 994/1009 Positives 997/1009 (98%) V.1 3 LLSGTYIFAVLLACVVFHSGAQEKNYTIREEMPENVLIGDLLKDLNLSLIPNKSLTTAMQ 62 LLSGTYIFAVLL CVVFHSGAQEKNYTIREE+PENVLIG+LLKDLNLSLIPNKSLTT MQ V.6 24 LLSGTYIFAVLLVCWVFHSGAQEKNYTIREEIPENVLIGNLLKDLNLSLIPNKSLTTTMQ 83 V.1 63 FKLVYKTGDVPLIRIEEDTGEI ETTGARIDREKLCAGIPRDEHCFYEVEVAILPDEIFRL 122 FKLVYKTGDVPLIRIEEDTGEI FTTGAIRIDREKLCAGIPRDEHCFYEVEVAILPDEIFRL V.6 84 FKLVYKTGDVPLIRIEEDTGEIFTTGARIDREKLCAGIPRDEHCFYEVEVAILPDEIFRL 143 V.1 123 VKIRFLIEDINDNAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVQNYELIKSQ 182

VKIRFLIEDINDNAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVQNYELIKSQ

V.6 ±144 VKIRFLIEDINDNAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVQNYELIKSO 203 V.1 183 NIFGLDVIETPEGDKI4PQLIVQKELDREEKDTYVZ4KVKVEDGGFPQRSSTAILQVSVTDT 242

NIFGLDVIETPEGDKMPQLIVQKELDREEKDTYVMKVKVEDGGFPQRSSTAILQVSVTDT

V.6 204 NIFGLDVIETPEGDKMPQLIVQKELDREEKDTYVMKVKVEDGGFPQRSSTAILQVSVTDT 263 V.1 243 NDN4HPVEXETEIEVSIPENAPVGTSVTQLHATDADIGENAKIHFSFSNLVSNIA.RLFHL 302 NDNHPVFKETEIEVSI PENAPVGTSVTQLHATDADIGENAIIHFSFSNLVSNIAPRLFHL V.6 264 NDNHPVFKETEIEVSIPENAPVGTSVTQLHATDADIGENAKIHFSFSNLVSNIARRLFHL 323 V.1I 303 NATTGLITIKEPLDREETPNHKLLVLASDGGLMPARAMVLVNVTDVNDNVPSIDIRYIVN 362

NAT'EGLITIKEPLDREETPNHKLLVLASDGGLMPAPAMVLVNVTDVNDNVPSIDIRYIVN

v.6 324 NATTGLITIKPLDEETPNHLLVLADGGMPAAMVLVNVTDVNDNVP3IDIRY~vN4 363 V. 1 3 63 PVNDTVVLSENIPLNTKIALITVTDKDADINGRVTCTDHEIPFRLRPVFSNQFLLETAj\ 422 PVNDTVVLSENI PLNTKIALITVTDKDADHNGRVTCFTDHEIPFRLRPVFSNQFLLE AA VM 384 PVNDTVVLSENIPLNTKIALITVTDKDADHNGRVTCFTDHEIPFRLRPVFSNQF.LENAA 443 V.1 423 YLDYESTKEYAIKLLAADAGKPPLNQSAMLFIKVKDENDNAPVFIQSFVTVSIPENNSPG 482 YLDYESTKEYAIKLLAADAGKPPLNQSAMLIFIKVKDENDNAPVrrQSFVTVSIPENNSPG V.6 444 YLDYESTKEYAIKLLAADAGKPPLNQSAMLFIKVKDENDNAPVFTQSFVTVSIPENNSPG 503 V.1 483 IQLTKVSAMDADSGPNAKINYLLGPDAPPEFSLDCRTGMLTVVKKDREKEDKYLFI'ILA 542 IQL KVSA DAflSGPNA4INYLLGPDAPPEFSLD RTGMLTVVKKLDREKEDKYLFTILA V.6 504 IQLMVSATDADSGPNAEINYLLGPDAPPEFSLDRIRTGMLTWVKDRKEDKYLF'TILA 563 V.1 543 KDNGVPPLTSNVTVFVSIIDQNDNSPVETHNEYNFYVPENLPHGTVGLIVTDPDYGDN 602 KDNGVPPLTSNVTVFVSIIDQNDNSPVF'THUEY FYVPENLPRHGTVGLITVTDPDYGDN V.6 564 KDNGVPPLTSNVTVFVSIIDQNDNSPVFrHNEYKFYVPENLPRHGTVGLITVTDPDYGDN 623 V.1 603 SAVTLSILDENDDFTIDSQTGVIRPNISFDREKQESYTFYVAEDGGRVSRSSSAKVTIN 662 SAVTLSILDENDDF'rIDSQTGVIRPNISFDREKQESYTFYVKAEDGGRVSRSSSAKVTIN V.6 624 SAVTLSILDENDDFrIDSQGVIRPNISFD.EKQESYTFYVKAEDGGRVSRSSSAKVTIN 683 V.1 663 VVDVNDNKPVFIVPPSNCSYELVLPSTNPGTVVFQVIAVDNDTGMNAEVRYSIVGGNTRD 722 VVDVNDNKPVFIVPP N SYELVLPSTNPGTVVFQVIAVDNDTGM1NAEVRYSIVGGNTRD V.6 684 VVDVNDNKPVFIVPPYNYSYELVLPSTNPGTVVFQVIAVDNDTGMNAEVRYSIVGGNTRD 743 V.1 723 LFAIDQETGNITLMEKCDVTDLGLHRVLVKANDLGPDSLFSVIVNLFVNESVTNATLI 782 LFAIDQETGNITLMEKCDVTDLGLHRVLVKANiDLGQPDSLFSVVIVNLFVNESVTNATLI V. 6 744 LFAIDQETGNITLMEKCDVTDILLHRVLVKANDLGQPDSLFSVVIVNLFVNESV'fNATLI 803 V.1 783 NELVRKSTEAPVTPNTEIADVSSPTSDYVKILVAAVAGTfl'VVVVIFITAVVRCRQAPHL 842 NELVRKS EAPWIPNTEIADVSSPTSDYVKILVAAVAGTITVVVVIFITAVVRCRQAPHL V.6 804 NELVRXSIEAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVVVIFITAVVRCRQAPHL 863 V.1 843 KAAQKNKQNSEWATPNPENRQMIMM CXKKHSPKNLLLNFVTIEETKADDVDSDGNR 902 KAAQKN QNSEWATPNPENRQMIMMKKKKKKKKHSPKNLLLNFVTIEETKADDVDSDGNR V.6: 864 KAAQKNMQNSEWATPNPENRQMIMMKKKKKKKSPKLLLNFVTIEETKADDVDSDGNR 923 V.1: 903 VTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLARjYKSASPQPFQIQPETPLNSiKiuI 962 VTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLARHYKSASPQPAFIQIQPETPLN KHHI V.6 :924 VTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLARHYKSASPQPAFQIQPETPLNLKIHI 983 V.1 :963 IQELPLDNTFVACDSISKCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP 1011

IQELPLDNTFVACDSISKCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP

V.6 :984 IQELPLDNTFVACDSISKCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP 1032 Table 1.11(f). Nucleotide sequence of transcript variant IO9Pi D4 v.7 (SEQ ID NO: 267) ggtggtccag ttttcagaat attaatagct gatagttgtt cttcttcaaa cagatcaatt attcattgcc tattttgtga aattatttgt tcaatggtgg tcatttatct ctaattaaca ttttgttaac CtCCtCttCt tgtccgggac aggagaaaaa tgaaagacct agctagtgta tcttcactac agcattgctt agatacgttt tacctccaaa cctttaataa attcttgttt accatgttta tcaaacttta ttttttcact aagagaataa tttgtaacaa attctcttta acacttttat ttattcttaa gagtgtcaat atgcatgttt tttggtcagt gtacattttc ctacaccatc taacttgtcg caagaccgga cggcgctcgc ttatgaagtg tctgatagaa gatatggaat gcagttatgt ttcttatcca ggcgttagtc ttagtccctc ttgatgccca ttgcatttta atacccttta actatcttgg aggtactctg tgtacgaatt tatgctaaca agggttggct gttgtgcggg gcggtcctgc cgagaagaaa ctgattccaa gatgtgccac attgatcgtg gaggttgcca gatataaatg acactcctga caatctgaaa aagaaaaatc acatcaaccc ctttataatg gagctgaaga aacccatgtt ttttccctta tatattaaag tgtcattttt cataatattt tctcatttac tcttaataat ttaatacaac tagtatgcgt ttccagaaaa acaagtcctt tgattcgaat agaaattatg ttttgccgga ataatgcacc aatatcctga gttgcttact ctctaatccc aacctttttt tgtactttat cttttcacat ctctcctctc ccaaacttct attccttgcc aatggactat ataacaaaga actattgaat tattatct gatactgtag gattcagaac tgattttaat ttcttcttcc aaactgtcac ggtgttccac cgtcctgata gacaactact tgaagaggat tgctggtatc tgaaatattt attgttccca tccttttatc tgtataaatt ataatgatta taaatatttt tatatattta gtatcttatt agatttatca ttaaaacagt tcttctctct aagtgtttgt tctggcgccc ggcaacttgt atgcagttca actggtgaga ccaagggatg agactggtta gcaacagtta 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 00 00 tcaacatatc aattccagag aactcggcta taaactctaa atatactctc ccagcggctq ttgatcctga tttttggcct aaaaggagtt gtggctttcc acaaccaccc taggcacttc tccacttctc caccactgg agttactggt atgttacaga tcaatgacac ctgtgacgga ttcctttcag ttgactatga ctcctttgaa cagttttcac agttgatgaa tgctaggccc tagtgaagaa ataatggggt atgacaatag caaggcatqg cagttacgct tcatccgacc aggctgagga ttgatgtcaa tggttctacc acactggcat ttgcaatcga ttggtttaca gtgttgtaat aactggtgcg cctcaccaac tcgttgtagt ctgctcagaa tgataatgat atgttgtcac cactagacct cacctactac cacagcctgc aagaactgcc gcagttcaga ctgtgtccgt agatgtaact tgatttcaaa aagcaaaaat ctggcaatgg catgctgttt tgtatacact tgttgaataa ctctctgaaa aaccaaaaca atcattttta gcctgtaatt gaccatcctg gcgtggtggc gaacccggga gacagagcaa taggttttta ttttggctgg tggatcacct cgtagqcata cgatgtcatt agatagggaa ti :aaagat cc agtctttaag agtgacacag tttcagcaat acttatcaca tttggcaagt tgtcaatgat agttgttctt taaggatgcg attaaggcca gtccacaaaa tcagtcagca ccagtctttc agtaagtgca tgatgctcca actagataga accaccctta cccagttttc tacagtagga ctccatttta aaatatttca tggtggtaga tgacaacaaa gtccactaat gaatgcagag ccaagaaaca cagagtgttg tgtcaatctg caaaagcatt tagtgactat tattttcatc aaacatgcag gaagaaaaag tattgaagaa tcctattgat tttcaagcct cttccaaatt tctcgataac tccctacagc acacaccaga ttctaggaac attaggctaa catcttaaaa aaatttaaaa ctctgtttgc tatggttaat ttaaaacatt ggtatccagg ctctggttca ttacccacca ccagcacttt gctaacatgg gggcgcctgt ggcggagctt gactctgtct tgttgattgt gtgtggtggc gaggtcagga Laacggagttc gaaacaccag gagaaggata agtactgcta gagacagaga ctccatgcca ctagtctcca atcaaagaac gatggtggat aatgtcccat tcagaaaata gaccataatg gtattcagta gaatatgcca atgctcttca gtaactgttt acggatgcag cctgaattca gaaaaagagg accagcaatg actcacaatg ctaatcactg gatgagaatg tttgatagag gtatcacgtt ccagttttca ccaggcacag gttcgttaca ggcaacataa gtcaaagcta ttcqtqaatg gaagcaccag gtcaagatcc actgctgtag aattctgaat aaaaagaaga actaaggcag ctagaagagc gacagccctg cagcctgaaa acctttgtgg gtttctgact ccgactgatt aacaaaattc gatcattaat atgatgtcct tttatggaag tctgaatcaa atatcagtca atctccagga gcaagagaca gtgttttgaa ctctgcttaa gggaggccga tgaaacccca agtcccagct gcagtgagcc caaaaagaaa actcatgctg tcatacctgt gttcaagacc aaaactacga aaggagacaa cctatgtgat ttttgcaagt ttgaagtcag cagatgctga acattgccag cactggatag tgatgccagc ccattgacat ttccactcaa gcagggtgac atcagttcct ttaaattact tcaaagtgaa ctattcctga acagtgggcc gcctggatcg ataaatattt tcacagtctt aatacaaatt taactgatcc atgacttcac aaaaacaaga cttcaagtgc ttgtccctcc tggtctttca gcattgtagg cattgatgga atgacttagg aqtcagtgac tgaccccaaa tggttgcagc taagatgtcg gggctacccc agaagcattc atgatgttga aaacaatggg atttggcccg ctcccctgaa cctgtgactc gtggctatcc ccaggacatg cattcccctt tttgtaatct agtgaacctt agacagtgca cagccatgat tgaaacatgc gtttggaagt tttttaagac aatattgact aagttgagtg ggcgggtgga tctccactaa actcgggagg gagatggcgc aaaatgttca ttccactcct aatcccagca agtctggcca actaattaag gatgccacaa gaaagtaaag aagtgttact tataccagaa cataggtgaa gagattattt ggaagaaaca aagagcaatg aagatacatc caccaaaatt atgcttcaca cctggagaat ggctgcagat agatgaaaat gaataactct taatgctgag tcgtacaggc attcacaatt tgtaagcatt ctatgtccca tgattatgga cattgattca atcttacact caaagtaacc ttacaactat ggtaattgct aggaaacaca gaaatgtgat acagcctgat caatgctaca tactgagata tgttgctggc ccaggcacca aaacccagaa ccctaagaac cagtgatgga aaagtacaat acactacaaa tttgaagcac tatctccaat agtgacaacc aactattgaa ccaaaaaatt agatttccca gtgctttctt gcgcaataac gtaatataag aattacttgc gagctgaact cccaaacaaa aacataatat ggccgggcgc tcacgaggtc aaatacaaaa ctgaggcagg cactgcactc gtgatagaaa tttaattatt ctttgggagg acat agtcaaaaca ctgattgttc gttgaagatg gatacaaatg aatgctcctg aatgccaaga cacctcaatg ccaaaccaca gtgctggtaa gtcaatcctg gctctcataa gatcatgaaa gcagcatatc gctggcaaac gacaatgctc cctggcatcc atcaattacc atgctqactg ctggcaaaag attgatcaga gaaaaccttc gacaattctg caaactqgtg ttctatgtaa ataaatgtgg tcttatgaat gttgacaatg agagatctgt gttacagacc tctctcttca ctgattaatg gctgatgtat accataactg caccttaagg aacaggcaga ctgctgctta aacagagtca tgggtaacta tctgcctctc cacatcatcc tgttcctcaa ttcgaggtac atctgcagtg tcaatgattg ttataaaagc tagctgtaat agagtactct gctgtcttgg cctgtctgat agccaaacta caaaaaacaa tgctgagaaa ggtggctcac aggatattga aattagctgg agaatggcgt cagcctgggt ataattttac aaaaagttat ccgaggcggg 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4964 282 00 C) Table 1.ll1(l). Nucleotide sequence alignment of 109P1 D4 M. (SEQ ID NO: 268) and 1 09P1 D4 v.7 (SEQ ID NO: 269) Score =5664 bits (2946), Expect =0.0Identilies 3000/3027 Strand =Plus Plus V.1 852 ttgttgtccgggacgtacattttcgcggtcctgctagcatgcgtggtgttccactctggc 911 V.7 837 ttgttgtccgggacgtacattttcgcggtcctgctagtatgcgtggtgttccactctggc 896 V.1 912 gcccaggagaaaaactacaccatccgagaagaaatgccagaaaacgtcctgataggcgac 971 V.7 897 gcccaggagaaaaactacaccatccgagaagaaattccagaaaacgtcctgataggcaac 956 riV.1 972 ttgttgaaagaccttaacttgtcgctgattccaaacaagtccttgacaactgctatgcag 1031 riV.7 957 ttgttgaaagaccttaacttgtcgctgattccaaacaagtccttgacaactactatgcag 1016 00 V.1 1032 ttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaagaggatactggt 1091 V.7 1017 ttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaagaggatactggt 1076 V.1 1092 gagatcttcactactggcgctcgcattgatcgtgagaaattatgtgctggtatcccaagg 1151 V.7 1.077 gagatcttcactaccggcgctcgcattgatcgtgagaaattatgtgctggtatcccaagg 1136 V.1 1152 gatgagcattgcttttatgaagtggaggttgccattttgccggatgaaatatttagactg 1211 V.7 1137 gatgagcattgcttttatgaagtggaggttgccattttgccggatgaaatat~tagactg 1196 V.1 1212 gttaagatacgttttctgatagaagatataaatgataatgcaccattgttcccagcaaca 1271 V.7 :1197 gttaagatacgttttctgatagaagatataaatgataatgcaccattgttcccagcaaca 1256 V.1 :1272 gttatcaacatatcaattccagagaactcggctataaactctaaatatactctcccagcg 1331 V.7 :1257 gttatcaacatatcaattccagagaactcggctataaactctaaatatactctcccagcg 1316 V.1 :1332 gctgttgatcctgacgtaggaataaacggagttcaaaactacgaactaattaagagtcaa 1391 V.7 :1317 gctgttgatcctgacgtaggcataaacggagttcaaaactacgaactaattaagagtcaa 1376 V.1 :1392 aacatttttggcctcgatgtcattgaaacaccagaaggagacaagatgccacaactgatt 1451 V.7 :1377 aacatttttggcctcgatgtcattgaaacaccagaaggagacaagatgccacaactgatt 1436 V.1 :1452 gttcaaaaggagttagatagggaagagaaggatacctacgtgatgaaagtaaaggttgaa 1511 V.7 .1437 gttcaaaaggagttagatagggaagagaaggatacctatgtgatgaaagtaaaggttgaa 1496 V.1 :1512 gatggtggctttcctcaaagatccagtactgctattttgcaagtgagtgttactgataca 1571 V.7 1497 gatggtggctttcctcaaagatccagtactgctattttgcaagtaagtgttactgataca 1556 V.1 1572 aatgacaaccacccagtctttaaggagacagagattgaagtcagtataccagaaaatgct 1631 V.7 1557 aatgacaaccacccagtctttaaggagacagagattgaagtcagtataccagaaaatgct 1616 283 00 V.1 1632 cctgtaggcacttcagtgacacagctccatgccacagatgctgacataggtgaaaatgcc 1691 V.7 1617 cctgtaggcacttcagtgacacagctccatgccacagatgctgacataggtgaaaatgcc 1676 1692 aagatccacttctctttcagcaatctagtctccaacattgccaggagattatttcacctc 1751 V.7 1677 aagatccacttctctttcagcaatctagtctccaacattgccaggagattatttcacctc 1736 V.1 1752 aatgccaccactggacttatcacaatcaaagaaccactggatagggaagaaacaccaaac 1811 V.7 1737 aatgccaccactggacttatcacaatcaaagaaccactggatagggaagaaacaccaaac 1796 V. 82ccatacgttgcatagggatagcgagacagtcg17 V.1 1812 cacaagttactggttttggcaagtgatggtggattgatgccagcaagagcaatggtgctg 1871 00 V.1 1872 gtaaatgttacagatgtcaatgataatgtcccatccattgacataagatacatcgtcaat 1931 c-1V.7 1857 gtaaatgttacagatgtcaatgataatgtcccatccattgacataagatacatcgtcaat 1916 V.1 1932 cctgtcaatgacacagttgttctttcagaaaatattccactcaacaccaaaattgctctc 1991 V.7 1917 cctgtcaatgacacagttgttctttcagaaaatattccactcaacaccaaaattgctctc 1976 V.1 1992 ataactgtgacggataaggatgcggaccataatggcagggtgacatgcttcacagatcat 2051 V.7 1977 ataactgtgacggataaggatgcggaccataatggcagggtgacatgcttcacagatcat 2036 V.1 2052 gaaatccctttcagattaaggccagtattcagtaatcagttcctcctggagactgcagca 2111 H111il lli ii111 I 1111111 111 111 V.7 2037 gaaattcctttcagattaaggccagtattcagtaatcagttcctcctggagaatgcagca 2096 V.1 2112 tatcttgactatgagtccacaaaagaatatgccattaaattactggctgcagatgctggc 2171 V.7 2097 tatcttgactatgagtccacaaaagaatatgccattaaattactggctgcagatgctggc 2156 V.1 2172 aaacctcctttgaatcagtcagcaatgctcttcatcaaagtgaaagatgaaaatgacaat 2231 V.7 2157 aaacctcctttgaatcagtcagcaatgctcttcatcaaagtgaaagatgaaaatgacaat 2216 V.1 2232 gctccagjttttcacccagtctttcgtaactgtttctattcctgagaataactctcctggc 2291 V.7 2217 gctccagttttcacccagtctttcgtaactgtttctattcctgagaataactctcctggc 2276 V. 1 2292 atccagttgacgaaagtaagtgcaatggatgcagacagtgggcctaatgctaagatcaat 2351 V.7 2277 atccagttgatgaaagtaagtgcaacggatgcagacagtgggcctaatgctgagatcaat 2336 V.1 2352 tacctgctaggccctgatgctccacctgaattcagcctggattgtcgtacaggcatgctg 2411 V.7 2337 tacctgctaggccctgatgctccacctgaattcagcctggatcgtcgtacaggcatgctg 2396 V.1 2412 actgtagtgaagaaactagatagagaaaaagaggataaatatttattcacaattctggca 2471 V.7 2397 actgtagtgaagaaactagatagagaaaaagaggataaatatttattcacaattctggca 2456 00 V.1 2472 aaagataacggggtaccacccttaaccagcaatgtcacagtctttgtaagcattattgat 2531 V.7 2457 aaagataatggggtaccacccttaaccagcaatgtcacagtctttgtaagcattattgat 2516 V.1 2532 cagaatgacaatagcccagttttcactcacaatgaatacaacttctatgtcccagaaaac 2591 V.7 2517 cagaatgacaatagcccagttttcactcacaatgaatacaaattctatgtcccagaaaac 2576 V.1 2592 cttccaaggcatggtacagtaggactaatcactgtaactgatcctgattatggagacaat 2651 V.7 2577 cttccaaggcatggtacagtaggactaatcactgtaactgatcctgattatggagacaat 2636 2652 tctgcaqttacgctctccattttagatgagaatgatgacttcaccattgattcacaaact 2711 00 V.7 :2637 tctgcagttacgctctccattttagatgagaatgatgacttcaccattgattcacaaact 2696 V.1 :2712 ggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatcttacactttctat 2771 V.7 :2697 ggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatcttacactttctat 2756 V.1 :2772 gtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaagtaaccataaat 2831 V.7 :2757 gtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaagtaaccataaat 2816 V.1 2832 gtggttgatgtcaatgacaacaaaccagttttcattgtccctccttccaactgttcttat 2891 V.7: 2817 gtggttgatgtcaatgacaacaaaccagttttcattgtccctccttacaactattcttat 2876 V.1 :2892 gaattggttctaccgtccactaatccaggcacagtggtctttcaggtaattgctgttgaC 2951 V.7 :2877 gaattggttctaccgtccactaatccaggcacagtggtctttcaggtaattgctgttgaC 2936 V.1: 2952 aatgacactggcatgaatgcagaggttcgttacagcattgtaggaggaaacacaagagat 3011 V.7 2937 aatgacactggcatgaatgcagaggttcgttacagcattgtaggaggaaacacaagagat 2996 V.1 3012 ctgtttgcaatcgaccaagaaacaggcaacataacattgatggagaaatgtgatgttaca 3071 V.7 2997 ctgtttgcaatcgaccaagaaacaggcaacataacattgatggagaaatgtgatgttaca 3056 V.1 3072 gaccttggtttacacagagtgttggtcaaagctaatgacttaggacagcctgattctctc 3131 V.7 3057 gaccttggtttacacagagtgttggtcaaagctaatgacttaggacagcctgattctctc 3116 V.1 3132 ttcagtgttgtaattgtcaatctgttcgtgaatgagtcggtgaccaatgctacactgatt 3191 V.7 3117 ttcagtgttgtaattgtcaatctgttcgtgaatgagtcagtgaccaatgctacactgatt 3176 V.1 3192 aatgaactggtgcgcaaaagcactgaagcaccagtgaccccaaatactgagatagctgat 3251 V.7 3177 aatgaactggtgcgcaaaagcattgaagcaccagtgaccccaaatactgagatagctgat 3236 V.1 3252 gtatcctcaccaactagtgactatgtcaagatcctggttgcagctgttgctggcaccata 3311 V.7 3237 gtatcctcaccaactagtgactatgtcaagatcctggttgcagctgttgctggcaccata 3296 00 V.1 :3312 actgtcgttgtagttattttcatcactgctgtagtaagatgtcgccaggcaccacacctt 3371 V.7? 3297 actgtcgttgtagttattttcatcactgctgtagtaagatgtcgccaggcaccacacctt 3356 V.1 :3372 aaggctgctcagaaaaacaagcagaattctgaatgggctaccccaaacccagaaaacagg 3431 V.7 :3357 aaggctgctcagaaaaacatgcagaattctgaatgggctaccccaaacccagaaaacagg 3416 V.1 :3432 cagatgataatgatgaagaaaaagaaaaagaagaagaagcattcccctaagaacttgctg 3491 V.7 :3417 cagatgataatgatgaagaaaaagaaaaagaagaagaagcattcccctaagaacctgctg 3476 V.1 :3492 cttaattttgtcactattgaagaaactaaggcagatgatgttgacagtgatggaaacaga 3551 001111 1111iiiiiili l11liliHIM 111111111 00V.7 :3477 cttaatgttgtcactattgaagaaactaaggcagatgatgttgacagtgatggaaacaga 3536 V.1 :3552 gtcacactagaccttcctattgatctagaagagcaaacaatgggaaagtacaattgggta 3611 V.7 :3537 gtcacactagaccttcctattgatctagaagagcaaacaatgggaaagtacaattgggta 3596 V.1 :3612 actacacctactactttcaagcccgacagccctgatttggcccgacactacaaatctgcc 3671 V.7 :3597 actacacctactactttcaagcctgacagccctgatttggcccgacactacaaatctgcc 3656 V.1 3672 tctccacagcctgccttccaaattcagcctgaaactcccctgaattcgaagcaccacatc 3731 V.7 3657 tctccacagcctgccttccaaattcagcctgaaactcccctgaatttgaagcaccacatc 3716 V.1 3732 atccaagaactgcctctcgataacacctttgtggcctgtgactctatctccaagtgttcc 3791 111111111111111111111 ii 111111111 HIM111 V.7 3717 atccaagaactgcctctcgataacacctttgtggcctgtgactctatctccaattgttcc 3776 V.1 3792 tcaagcagttcagatccctacagcgtttctgactgtggctatccagtgacgaccttcgag 3851 V.7 3777 tcaagcagttcagatccctacagcgtttctgactgtggctatccagtgacaaccttcgag 3836 V.1 3852 gtacctgtgtccgtacacaccagaccg 3878 V.7 :3837 gtacctgtgtccgtacacaccagaccg 3863 Score =1567 bits (815), Expect 0.Oldentities 829/836 Strand -Plus/ Plus V.1 :3 qgtggtccagtacctccaaagatatggaatacactcctgaaatatcctgaaaactttttt 62 V.7 ;1 ggtggtccagtacctccaaagatatggaatacactcctgaaatatcctgaaacctttttt V.1 :63 ttttcagaatcctttaataagcagttatgtcaatctgaaagttgcttacttgtactttat 122 V.7 :61 ttttcagaatcctttaataagcagttatgtcaa~ctgaaagttgcttacttgtactttat 120 V.1 :123 attaatagctattcttgtttttcttatccaaagaaaaatcctctaatccccttttcacat 182 V.7 :121 attaatagctattcttgtttttcttatccaaagaaaaatcctctaatccccttttcacat 180 00 V.1 :183 gatagttgttaccatgtttaggcattagtcacatcaacccctctcctctcccaaacttct 242 V.7 :181 gatagttgttaccatgtttaggcgttagtcacatcaacccctctcctctcccaaacttct 240 V.1 :243 cttcttcaaatcaaactttattagtccctcctttataatgattccttgcctcgttttatc 302 V.7 :241 cttcttcaaatcaaactttattagtccctcctttataatgattccttgcctccttttatc 300 V.1 :303 cagatcaattttttttcactttgatgcccagagctgaagaaatggactactgtataaatt 362 V.7 :301 cagatcaattttttttcactttgatgcccagagctgaagaaatggactattgtataaatt 360 rlV.1 :363 attcattgccaagagaataattgcattttaaacccatattataacaaagaataatgatta 422 V.7 :361 attcattgccaagagaataattgcattttaaacccatgttataacaaagaataatgatta 420 00 V.1 :423 tattttgtgatttgtaacaaataccctttattttcccttaactattgaattaaatatttt 482 V.7 :421 tattttgtgatttgtaacaaataccctttattttcccttaactattgaattaaatatttt 480 V.1 483 aattatttgtattctctttaactatcttggtatattaaagtattatcttttatatattta 542 V.7 481 aattatttgtattctctttaactatcttggtatattaaagtattatcttttatatattta 540 V.1 543 tcaatggtggacacttttataggtactctgtgtcatttttgatactgtaggtatcttatt 602 V.7 :541 tcaatggtggacacttttataggtactctgtgtcatttttgatactgtaggtatcttatt 600 V.1 ;603 tcatttatctttattcttaatgtacgaattcataatatttgattcagaacaaatttatca 662 V. 7 :601 tcatttatctttattcttaatgtacgaattcataatatttgattcagaacagatttatca 660 V.1 :663 ctaattaacagagtgtcaattatgctaacatctcatttactgattttaatttaaaacagt 722 V.7 :661 ctaattaacagagtgtcaattatgctaacatctcatttactgattttaatttaaaacagt 720 V.1 :723 ttttgttaacatgcatgtttagggttggcttcttaataatttcttcttcctcttctctct 752 V.7 :721 ttttgttaacatgcatgtttagggttggcttcttaataatttcttcttcctcttctctct 780 V.1 :783 ctcctcttcttttggtcagtgttgtgcgggttaatacaacaaactgtaacaagtgt 838 V.7 :781 ctcctcttcttttggtcagtgttgtgcgggttaatacaacaaactgtcacaagtgt 836 Table UV(f). Peptide sequences of protein coded by 109P1D4 v.7 (SEQ ID NO: 270) MFRVGFLIIS SSSSLSPLLL VSVVRVNTTN CHKCLLSGTY IFAVLLVCVV FHSGAQEKNY TIREEIPENV LIGNLLKDLN LSLIPNKSLT TTMQFKLVYK TGDVPLIRIE EDTGEIFTTG 120 ARIDREKLCA GIPRDEHCFY EVEVAILPDE IFRLVKIRFL IEDINDNAPL FPATVINISI 1830 PENSAINSKY TLPAAVDPDV GINGVQNYEL IKSQNIFGLD VIETPEGDKM PQLIVQKELD 240 REEKDTYVMK VKVEDGGFPQ RSSTAILQVS VTDTNDNHPV FKETEIEVSI PENAPVGTSV 300 TQLHATDADI GENAKIHFSF SNLVSNIARR LFHLNATTGL ITIKEPLDRE ETPNHKLLVL 360 ASDGGILMPAR AMVLVNVTDV NDNVPSIDIR YIVNPVNDTV VLSENIPLNT KIALITVTDK 420 DADHNGRVTC FTDHEIPFRL RPVFSNQFLL ENAAYLDYES TKEYAIKLLAi ADAGKPPLNQ 480 SAMLFIKVKD ENDNAPVFTQ SFVTVSIPEN NSPGIQLMKV SATDADSGPN AEINYLLGPD 540 APPEFSLDRR TGMLTVVKKL DREKEDKYLF TII.AKDNGVP PLTSNVTVFV SIIDQNDNSP 600 VFTHNEYKFY VPENLPRHGT VGLITVTDPD YGDNSAVTLS ILDENDDFTI DSQTGVIRPN 660 00ISFDREKQES YTFYVKAEDG GRVSRSSSAK VTINVVDVND NKPVFIVPPY NYSYELVLPS 720 TNPGTVVFQV IAVDNDTGMN AEVRYSIVGG NTRDLFAIDQ ETGNITLMEK CDVTDLGLHR 780 VLVKANDLGQ PDSLFSVVIV NLFVNESVTN ATLINELVRK SIEAPVTPNT EIADVSSPTS 840 DYVKILVAAV AGTITVVVVI FITAVVRCRQ APHLKAAQKN MQNSEWATPN PENRQMIMMK 900 KKKKKKKHSP KNLLLNVVTI EETKADDVIDS DGNRVTLDLP IDLEEQTMGK YNWVTTPTTF 960 KPDSPDLARH YKSASPQPAF QIQPETPLNL KHHIIQELPL DNTFVACDSI SNCSSSSSDP 1020 YSVSDCGYPV TTFEVPVSVH TRPTIDSRT 1048 Table LV(tQ. Amino acid sequence alignment of 1O9PI D4 v. (SEQ ID NO: 271) and I09PI D4 v.7 (SEQ ID NO: 272) Score =1961 bits (5081), Expect 0.Oldentities 992/1009 Positives 995/1009 (98%) V.1 3 LLSGTYIFAVLLACVVFHSGAQEKNYTIREEMPENVLIGDLLKDLNLSLI PNKSLTTAMQ 62 LLSGTYIFAVLL CVVFHSGAQEKNYTIREE+FENVLIG+LLIKDLNLSLI PNKSLTT MQ v.7 35 LLSGTYIFAVLLVCVVFHSGAQEKNYTIREEIPENVLIGNLLKDLNLSLIPNKSLTTTMQ 94 63 FKLVYKTGDVPLIRIEEDTGEIFTTGARIDREKLCAGIPRDEHCFYEVEVAILPDEIFRL 122

FKLVYKTGDVPLIRIEEDTGEIFTTGARIDREKLCAGIPRDEHCFYEVEVAILPDEIFRL

00 V.7 95 FKLVYKTGDVPLIRIEEDTGEIFrTGAP.IDREKLCAGIPRDEHCFYEVEVAILPDEIFRL 154 V.1 123 VKIRFLIEDINDNAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVQNYELIKSQ 182

VKIRFLIEDINDNAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVQNYELIKSQ

V.7 155 VKIRFLIEDINDNAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVQNYELIKSQ 214 V.1 183 NIFGLDVIETPEGDKMPQLIVQKELDREEKDTYVMKVKVEDGGFPQRSSTAILQVSVTDT 242

NIFGLDVIETPEGDKMPQLIVQKELDREEKDTYVMKVKVEDGGFPQRSSTAILQVSVTDT

V.7 215 NIFGLDVIETPEGDKMPQLIVQKELDREEKDTYVMKVKVEDGGFPQRSSTAILQVSVTDT 274 V.1 243 NDNHPVFKETEIEVSIPENAPVGTSVTQLHATDADIGENAKIHFSFSNLVSNIARRL'HL 302 NDNHPVFKETEIEVSI PENAPVGTSVTQLHATDADIGENAXIHFSFSNLVSNIAR&LFHL V.7 275 NDNHPVFKETEIEVSIPENAPVGTSVTQLHATDADIGENAKIHFSFSNLVSNIARP.LFHL 334 V.1 303 NATTGLITIKEPLDREETPNHKLLVLASDGGLMPARAMVLVNVTDVNDNVPSIDIRYIVN 362

NATTGLITIKEPLDREETPNHKLLVLASDGGLMPARAMJVLVNVTDVNDNVPSIDIRYIVN

V.7 335 NATTGLITIKEPLDREETPNHKLLVLASDGGL4PARAMVLVNVTDVNDNVPSIDIRYIV4 394 V.1 363 PVNDTVVLSENIPLNTKIALITVTDKDADHNGRVTCFTDHEIPFRLRPVFSNQFLLETAA 422 PNDTVVLSENIPLNTKIALITVTDKDADHNGRVTCFTDHEIPFRLRPVFSNQFLLE A V.7 395 PVNDTVVLSENIPLNTKIALITVTDKDADHNGRVTCFTDHEIPFRLRPVFSNQFLLENAA 454 V.1 423 YLDYESTKEYAIKLLAADAGKPPLNQSAMLFIKVKDENDNAPVFrQSFVTVSIPENNSPG 482 YLDYESTKEYAIKLLAADAGKPPLNQSAI4LFIKVKDENDNAPVFTQSFVTVSIPENNSPG V.7 455 YLDYESTKEYAIKLLAADAGKPPLNQSAMLFIKVKDENDNAPV~rQSFVTVSIPENNSPG 514 V.1 483 IQLTKVSAMDADSGPNAKINYLLGPDAPPEFSLDCRTGMLTVVKKLDREKEDKYLFTILA 542 IQL KVSA DADSGPNA+INYLLGPDAPPEFSLD RTGMLTVVKKLDREKEDKYLFTILA V.7 515 IQLI4 VSATDADSGPNAEINYLLGPDAPPEFSLDRRTGMLTVVKLDREKEDKYL~FTLA 574 V.1 543 KDNGVPPLTSNVTVFVSIIDONDNSPVFTHNEYNFYVPENLPRHGTVGLITVTDPDYGDN 602 KDNGVPPLTSNVTVFVSII DQNDNSPVFTHNEY FYVPENLPRHGTVGLITVTDPDYGDN V.7 575 KDNGVPPLTSNVTVFVSIIDQNDNSPVFTHNEYKFYVPENLPRHGTVGLITVTDPDYGDN 634 V.1 603 SAVTLSILDENDDFTIDSQTGVIRPNISFDREKQESYTFYVKAEDGGRVSRSSSAKVTIN 662

SAVTLSILDENDDFTIDSQTGVIRPNISFDREKQESYTFYVKAEDGGRVSRSSSAKVTIN

V.7 635 SAVTLSILDENDDFTIDSQTGVIRPNISFDREKQESYTFYVKAEDGGRVSRSSSAKVTIN 694 V.1 663 VDVDNKPVFIVPPSNCSYLVLPSTNPGTVVFQVIAVDN.DTGMNAEVRYSIVGGNTRD 722 VVDVNDNKPVFIVPP N SYELVLPSTNPGTVVFVIAVDNDTGMNAEVRYSIVGGNTRD V.7 695 VVDVNDNKPVFIVPL'YNYSYEILVLPSTNPGTVVFQVIAVDNDTGMNAEVRYSIVGGNTRD 754 V.1 723 LFAIDOETGNITLMEKCDVTDLGLHRVLVKANDLGQPDSLFSVVIVNLaFVNESVTNATLI 782

LFAIDOETGNITLMEKCDVTDLGLHRVLVKANDILGPDSLFSVVIVNLFVNESVTNATLI

V.7 755 LFAIDQETGNITLMEKCDVTDLGLHRVLVKANDLGQPDSLFSVVIVNLFVNESVTNATLI 814 V.1 783 NELVRKSTEAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVVVIFITAVVRCRQAPHL 842 NELVRKS EAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVVVIFITAVVRCRQAPHL V.7 815 NELVRXSIEAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVVVIFITAVVRCRQAPHL 874 VA1 843 KAAQKNKQNSEWATPNPENR HIMM KKKKKKKHSPKNLLLNFVITIEETKADDVDSDGNR 902 00 KAAQKN QNSEWATPNPENRQMIMMKKKKKKKKHSPKNLLLN VTIEETKADDVDSDGNR V.7 875 KAAQKNMQNSEWATPNPENRQMIMMKKKKKKKKHSPKNLLLNVVTIEETKADDVDSDGNR 934 v.1 903 VTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLZ RHYKSASPQPAFQIQPETPLNSKHI 962 VTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLARHYKSASPQPAFQIQPETPLN KHHiI V.7 935 VTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLARHYKSASPQPAFQIQPETPLNLKHHI 994 V.1 963 IQELPLDNTFVACDSISKCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP 1011 IQEIJPLDNTFVACDSIS CSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP V.7 995 I0ELPLDNTFVACDSISNCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP 1043 Table UI(g). Nucleotide sequence of transcipt variant I 09P1 D4 v.8 (SEQ ID NO: 273) ggtggtccag ttttcagaat attaatagct gatagttgtt cttcttcaaa cagatcaatt attcattgcc tattttgtga aattatttgt tcaatggtgg tcatttatct ctaattaaca ttttgttaac ctcctcttct tgtccgggac aqgagaaaaa tgaaagacct agctagtgta tcttcactac agcattgctt agatacgttt tcaacatatc ttgatcctga tttttggcct aaaaggagtt gtggctttcc acaaccaccc taggcacttc tccacttctc ccaccactgg agttactggt atgttacaga tcaatgacac ctgtgacgga ttcctttcag ttgactatga ctcctttgaa cagttttcac agttgatgaa tgctaggccc tagtgaagaa ataatggggt atgacaatag caaggcatgg cagttacgct tcatccgacc aggctgagga ttgatgtcaa tgqttctacc acactggcat tacctccaaa cctttaataa attcttgttt accatgttta tcaaacttta ttttttcact aagagaataa tttgtaacaa attctcttta acacttttat ttattcttaa gagtgtcaat atgcatgttt tttggtcagt gtacattttc ctacaccatc taacttgtcg caagaccgga cggcgctcgc ttatgaagtg tctgatagaa aattccagag cgtaggcata cgatgtcatt agatagggaa tcaaagatcc agtctttaag agtgacacag tttcagcaat acttatcaca tttggcaagt tgtcaatgat agttgttctt taaggatgcg attaagqcca gtccacaaaa tcagtcagca ccagtctttc agtaagtgca tgatgctcca actagataga accaccctta cccagttttc tacagtagga ctccatttta aaatatttca tggtggtaga tgacaacaaa gtccactaat gaatgcagag gatatggaat gcagttatgt ttcttatcca ggcgttagtc ttagtccctc ttgatgccca ttgcatttta atacccttta actatcttgg aggtactctg tgtacgaatt tatgctaaca agggttggct gttgtgcggg gcggtcctgc cgagaagaaa ctgattccaa gatgtgccac attgatcgtg gaggttgcca gatataaatg aactcggcta aacggagttc gaaacaccag gagaaggata agtactgcta gagacagaga ctccatgcca ctagtctcca atcaaagaac gatggtggat aatgtcccat tcagaaaata gaccataatg gtattcagta gaatatgcca atgctcttca gtaactgttt acggatgcag cctgaattca gaaaaagagg accagcaatg actcacaatg ctaatcactg gatgagaatg tttgatagag gtatcacgtt ccagttttca ccaggcacag gttcgttaca acactcctga caatctgaaa aagaaaaatc acatcaaccc ctttataatg gagctgaaga aacccatgtt ttttccctta tatattaaag tgtcattttt cataatattt tctcatttac tcttaataat ttaatacaac tagtatgcgt ttccagaaaa acaagtcctt tgattcgaat agaaattatg ttttgccgga ataatgcacc taaactctaa aaaactacga aaggagacaa cctatgtgat ttttgcaagt ttgaagtcag cagatgctga acattgccag cactggatag tgatgccagc ccattgacat ttccactcaa gcagggtgac atcagttcct ttaaattact tcaaagtgaa ctattcctga acagt gggcc gcctggatcg ataaatattt tcacagtctt aatacaaatt taactgatcc atgacttcac aaaaacaaga cttcaagtgc ttgtccctcc tggtctttca gcattgtagg aatatcctga gttgcttact ctctaatccc ctctcctctc attccttgcc aatggactat ataacaaaga actattgaat tattatcttt gatactgtag gattcagaac tgattttaat ttcttcttcc aaactgtcac ggtgttccac cgtcctgata gacaactact tgaagaggat tgctggtatc tgaaatattt attgttccca atatactctc actaattaag gatgccacaa gaaagtaaag aagtgttact tataccagaa cataggtgaa gagattattt ggaagaaaca aagagcaatg aagatacatc caccaaaatt atgcttcaca cctggagaat ggctgcagat agatgaaaat gaataactct taatgctgag tcgtacaggc attcacaatt tgtaagcatt ctatgtccca tgattatgga cattgattca atcttacact caaagtaacc ttacaactat ggtaattgct aggaaacaca aacctttttt tgtactttat cttttcacat ccaaacttct tccttttatc tgtataaatt ataatgatta taaatatttt tatatattta gtatcttatt agatttatca ttaaaacagt tcttctctct aagtgtttgt tctggcgccc ggcaacttgt atgcagttca actggtgaga ccaagggatg agactggtta gcaacagtta ccagcggctg agtcaaaaca ctgattgttc gttgaagatg gatacaaatg aatgctcctg aatgccaaga cacctcaatg ccaaaccaca gtgctggtaa gtcaatcctg gctctcataa gatcatgaaa gcagcatatc gctggcaaac gacaatgctc cctggcatcc atcaattacc atgctgactg ctggcaaaag attgatcaga gaaaaccttc gacaattctg caaactggtg ttctatgtaa ataaatgtgg tcttatgaat gttgacaatg agagatctgt 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 00 00 ttgcaatcga ttggtttaca gtgttgtaat aactggtgcg cctcaccaac tcgttgtagt ctgctcagaa tgataatgat atgttgtcac cactagacct cacctactac cacagcctgc aagaactgcc gcagttcaga ctgtgtccgt ctcaggaaag catcccatgg gcaatcgcac agaaagaaat gtctcatcta cttcacaagc agcaccacag agaccatcgc ctcctctagt tctgctacag aggttattgc tgcaaggtgc agttttacac tgacaacctt aaacacatcc tcaaaattta taaatagaaa aaaattgcaa gcatcccctt tttttgttgt gaaccagtat tttacctaaa gctttctttt cttcaaaata tttctacctt aatatgtata tcgagacaca tttttacttt agcctgtagt aaaattaaaa tactattttg ttaccaaagg taaatctata aataacaaca aacagaattt caagaagaaa aaaatatatt gttgtctgtt ttataaatgt ccttgtttca ttggggggga ttgatattta tttcattggt ttaagtagcg tttgactggg ttaattaatt ttttccaggt ccaagaaaca cagagtgttg tgtcaatctg caaaagcatt tagtgactat tattttcatc aaacatgcag gaagaaaaag tattgaagaa tcctattgat tttcaagcct cttccaaatt tctcgataac tccctacagc acacaccaga cagcagtgat cctgcccctt tgaaggggat aactgttcaa tggccattct acaggcctct cccaccagtg attgtgccac gcagggtact ccctccttta cctccatcgt taatggacta catgtctgaa cgctccacgc cttgtaaagc agatacaatt cagataccag tttgtttaat gtacagtaag ttttatcatc gtagcaaatg cttttgttca tgatttttqt cacagtaggt attccaatac aactgtacag gaagtgcaat aatagtttag ccattattac ggaggtggat tagagaaact gtgttcagta agactaaggg agaatatcag taaggaaaaa taccaataac cttctaagca atttccttct acagaaacat gtataacact gtcaatatct atttatgatt gaggatttcc ttttcagaat cgtgcagaat ttttttctcc gcatgtgaat *ggcaacataa gtcaaagcta ttcgtqaatg gaagcaccag gtcaagatcc actgctgtag aattctgaat *aaaaagaaga actaaggcag ctagaagagc gacaqccctq cagcct gaaa acctttgtgg gtttctgact ccgtcccagc ggtggactgg ggctatcctc ggcaactccg ccaactgtgg gatgcctgct gctctatgcc acacagacca agcccaccac gcacttcacc gcacaggctg agtcaggccc tgctctgttg agacttcatc caacaggcca taaaatagtt ccaatgagta aataaatcta tcagaatgtg gcttatcatg atgtgcaata gaaagcctag gataatgtta ttgtggtttt agtgtaaata tatattgttg atctagatct aactgcccta tgtaaagtac ttgggtcttt gcatccaaag caggaagat t aaaataacaa atttttgtta gaaggaactt tgcagaggga acacacagaa aagaaacagt tttatcctct caccaacttg aaaccaagag cctattgatt aaactgtaat tttgaatatt atacattatt aacttcttaa ttcgtgttgt cccgaaggac cattgatgqa atgacttagg agtcagtgac tgaccccaaa tggttgcagc taagatgtcg gggctacccc agaagcattc atgatgttga aaacaatggg atttggcccg ctcccctgaa cctgtgactc gtggctatcc ggcgtgtcac gagaccatga aggaggagta atcctgaatc aagaggcctc ggatgccggc acagcccacc ttgttctctg cgatacaggt acagcccacc ctgcaatcag aatcatcagt atcagggagt ccagtgatga gaccgtccag acttcaaatt ttctgattat cagctagacc tatttaaaaa acagagcgta ttactgattt aaatatctta aaaggtatac cagttttttt ctgcttgttt ataaaatttg acaacctatt attaagcaac atcagaaata acttctggga cacgagtcac taaatgttga atacatgtaa ttctagctca ttcaagaaat gaaataaggc caaaaaccat actattcata taactggcca attttcttcc acaattgatg aacttagaca tttgtaacat gagaaagttt cccatccatt ctattaacta taatgttcca tgatgatatt qaaatgtga t acagcctgat caatgctaca tactgagata tgttgctggc ccaggcacca aaacccagaa ccctaagaac cagtgatgga aaagtacaat acactacaaa tttgaagcac tatctccaat agtgacaacc atttcacctg tgcaggcagc ctttgatcgt tactttcata tgacaactgc atctctggat actgtcacag ccacagccct gtctgctctc atcagcacag ccacagctct cagtttgcag gcaaggtagt ttcaattaaa aggtgattcc ttcagaaaag cagatttgta cttagtcaat gaaaaggaat ctatttctga gtttccatgc ttttctaagt qtactctagc gttgttgtta gtgtctctct tatatacatt tctctactct tatttgttaa aaactgtatc atttgtatgt ttaaaatatc tttgacagct ctgtagataa acttactgaa gtaattataa acatgactgc caaaatctca gaaaacatta ttatcttgta atagcaaaac tttaatgggg tagattttgt aaactgtggt cttttcatgt gtaaagttcc tcagagtttg agggatttgg tgaatgttta gttacaqacc tctctcttca ctgattaatg gctgatgtat accataactg caccttaagg aacaggcaga ctgctgctta aacagagtca tgggtaacta tctgcctctc cacatcatc tgttcctcaa ttcgaggtac ccagaaggct cttaccagca gctacaccca cctggactaa actcaagaat cattccagct gcctctactc ccagtgacac caccacagtc gcctcagccc tctctgccac caaggttggg gcaacatctc gtcattcctt cccattatgg atgtatatag aataactatg agttaaccaa ttaacaattt tgtacagtat tgattgtgtg ttacctttag cttttttggg gtgagtctcc gctgtcatgt ttcaataaag ttagtagagt aaagggcccc tgacatttta aacagcctag gacggtaaac caataggctg aaccacatac gaaaaccact atctacatca ttcttgcagt tatatgaaat gttttctcct tgtgcacatt tgagaaaata gcggttgggg aatgtataac aattgcataa gcccagcagg ttaagtcata attaataaaa agcatactgg ttaaattatt 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5760 5820 5880 5940 6000 6060 6120 6180 6240 6300 6360 6420 6480 6540 6600 6660 6720 00 00 atcacacaaa agattaagaa ctttgcctca cacattttct acacacacac tattaaataa gactcaaagt gaggttctta taqgaatgaa gattgtgttt ttgaatggaa ccctatgcct cggcttcaga gacagtggta cccacaactg gattttgttt tcaatcaaag tctagctatg tatgctacgt tcagtgtatt aattttgtgt aataataata tcccaaacta cacaaattgt tcctgagaaa tgtttctttt tctacttgtt tttcttatct ttattatgac catgggaaca ttaccacacc tacactaacc cagcagaaac ttaatggcaa ttatactatc gtgggaagat cagttccaag atgtaatcca tcttattaat tgtgttgata aagaaccaat aaaaacctgc ctctgcaaac acgtgcacac cttatcaggc tggatatttg aatcctcata ccaatacgct agaatattaa gcagqctgag tctcttccta tctgatgata taggttgaca caaaacgtct gatttgaaat tagttattct gtattactat gcatacacat aa aa at tagc attcttattt aatggttagt atatttatca tcagttctta attaaaggtg tcatggtatt tagactttta tcttttatat cattagaaac agagaagctg agatcaattt gtcatgtgct agtaacaaaa ttqtttagtg cctgcttaaa tcctcctctg catgctttgg gaagcaaact atatattaat ttgtggctat agtgacaaaa aaagatgatg ctttagtttt acacacattt atctcaatgg ggatttttct tctggaaact tttattaccc atgactgggc agtagccaaa agcgtccact tctttttcat cagcacacaa tactgaagca taaaacttca attttgtgtc atcacacttg tcttttctta ttttacatat acttaacatt gctattgtgt cacatggtca aaatgtaatt ttttactcac actgaaggtg tttatactct tttttggaaa tattttgaat aaatatattt tatgcagagg tttagtaaat aaatctgctc actgtaagtg atattaagat tgatatcata aagtaagaag tgtattaatt aaaagttatg tgttgatgtt atcagtgctt tgagattttt ctgatgatct aaaggatata ttactatcta tctgacagta tgtgaagttt tttctaactc accctcttct gaggcaaggg aggtctggcc cacattacaa gtggctattg acaatcgaaa aactgaatga catattccat tgagtatgta aactttacct gatatctaca ttacttttaa aatggtagca ttaaatggga atgtcacaca atggatattt tgtatactcc tctgatttat ccaaatttat gtttccaact ctgcaagaac ccttaaaata atgattttta agaaaaatgt atacttttta ttttatgaaa cagcatctga gttgactatt gttctatttc agaaat gaaaattgta ccagtagatt tcttgtgttt acacacacac aaaagaagag tgttagtgaa taatttattg tgacaccttt tgattttata tggtttttac gtattagccc ttggaaatct gttatttctt tgatgtatga aatggttctg cttatatgag tagattgtga ttcaaatact gtgttttaac atgtaataaa ttatgtaaat gttacaaaga aaaaaataga cacaaaaaaa caacattagt ctaatacaca attttttatt agttagttta ggctcaattg ctttctatat ttctttcaca aaagcagttc atgtgcacaa aagagtaaac tatgtattta aagtgaacag gtatggccaa aggttctgta aacttqggqa ttagaacatt taattatttt atacacacac gttgaaagat aatcaaatag agttactagg cctatagata atcagactta cagagaggct agttattctc gttacttcta tgactgaata tgtatgtagt ttttaaaaag aataatatgt ttattaattt aagtatctta taatattgtg tttagagagt ttggttagaa gcctctgcct ctaaacaaat tccttttcaa tttttttgtt tttatgaaaa ataattatta ggtaaacttt gctgggaaaa tatgtgccaa gtagctttct aagttgacaa ataaaaaaaa tgtgtgaaat tgtttgtatt tatcccaaag ggatggcagt ttgcatgttt 6780 6840 6900 6960 7020 7080 7140 7200 7260 7320 7380 7440 7500 7560 7620 7680 7740 7800 7860 7920 7980 8040 8100 8160 8220 8280 8340 8400 8460 8520 8580 8640 8700 8760 8820 8880 8940 9000 9036 Table 1.I11(g). Nucleotide sequence alignment of 1O9P1D4v.1 (SEQ ID NO: 274)and 1O9PID4v.S (SEQ ID NO: 275) Sore 5664 bits (2946), Expect =0.Oldentities =3000/3027 Stand Plus Plus V.1 852 ttgttgtccgggacgtacattttcgcggtcctgctagcatgcgtggtgttcactctggc 911 V.8 837 ttgttgtccgggacgtacattttcgcggtcctgctagtatgcgtggtgttccactctggc 896 V.1 912 gcccaggagaaaaactacaccatccgagaagaaatgccagaaaacgtcctgataggcgac 971 V.8 897 gcccaggagaaaaactacaccatccgagaagaaattccagaaaacgtcctgataggcaac 956 V.1 972 ttgttgaaagaCCttaacttgtcgctgattccaaacaagtccttgacaactgctatgcag 1031 V.8 957 ttgttgaaagaccttaacttgtcgctgattccaaacaagtccttgacaactactatgcag 1016 V.1 1032 ttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaagaggatactggt 1091 V.8 1017 ttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaagaggatactggt 1076 V.1 1092 gagatcttcactactggcgctcgcattgatcgtgagaaattatgtgctggtatcccaagg 1151 00 V.8 :1077 gagatcttcactaccggcgctcgcattgatcgtgagaaattatgtgctggtatcccaagg 1136 V.1 :1152 gatgagcattgcttttatgaagtggaggttgccattttgccggatgaaatatttagactg 1211 V.8: 1137 gatqagcattgcttttatgaagtggaggttgccattttgccggatgaaatatttagactg 1196 V.1 :1212 gttaagatacgttttctgatagaagatataaatgataatgcaccattgttcccagcaaca 1271 V.8 :11.97 gttaagatacgttttctgatagaagatataaatgataatgcaccattgttcccagcaaca 1256 V.1 :1272 gttatcaacatatcaattccagagaactcggctataaactctaaatatactctcccagcg 1331 V.8 :1257 gttatcaacatatcaattccagagaactcggctataaactctaaatatactctcccagcg 1316 00V.1 :1332 gctgttgatcctgacgtaggaataaacggagttcaaaactacgaactaattaagagtcaa 1391 0l0ll~ll111 111 ii111 ii11111111 V.8 :1317 gctgttgatcctgacgtaggcataaacggagttcaaaactacgaactaattaagagtcaa 1376 V. 32actttgctgttataaccaagaaagtgccatat15 V.8 1 1392 aacatttttggcctcgatgtcattgaaacaccagaaggagacaagatgccacaactgatt 1451 V.1 :1452 gttcaaaaggagttagatagggaagagaaggatacctacgtgatgaaagtaaaggttgaa 1511 V.8 :1437 gttcaaaaggagttagatagggaagagaaggatacctatgtgatgaaagtaaaggttgaa 1496 V.1 1512 gatggtggctttcctcaaagatccagtactgctattttgcaagtgagtgttactgataca 1571 V.8 1497 gatggtggctttcctcaaagatccagtactgctattttgcaagtaagtgttactgataca 1556 V.1 1572 aatgacaaccacccagtctttaaggagacagagattgaagtcagtataccagaaaatgct 1631 V.8 1557 aatgacaaccacccagtctttaaggagacagagattgaagtcagtataccagaaaatgct 1616 V.1 :1632 cctgtaggcacttcagtgacacagctccatgccacagatgctgacataggtgaaaatgcc 1691 V.8 :1617 cctgtaggcacttcagtgacacagctccatgccacagatgctgacataggtgaaaatgcc 1676 V.1 :1692 aagatccacttctctttcagcaatctagtctccaacattgccaggagattatttcacctc 1751 V.8 :1677 aagatccacttctctttcagcaatctagtctccaacattgccaggagattatttcacctc 1736 V.1 :1752 aatgccaccactggacttatcacaatcaaagaaccactggatagggaagaaacaccaaac 1811 V.8 :1737 aatgccaccactggacttatcacaatcaaagaaccactggatagggaagaaacaccaaac 1796 V.1 :1812 cacaagttactggttttggcaagtgatggtggattgatgccagcaagagcaatggtgctg 1871 V.8 :1797 cacaagttactggttttggcaagtgatggtggattgatgccagcaagagcaatggtgctg 1856 V.1 :1872 gtaaatgttacagatgtcaatgataatgtcccatccattgacataagatacatcgtcaat 1931 V.8 :1857 gtaaatgttacagatgtcaatgataatgtcccatccattgacataagatacatcqtcaat 1916 V.1 :1932 cctgtcaatgacacagttgttctttcagaaaatattccactcaacaccaaaattgctctc 1991 292 00 V.8 :1917 cctgtcaatgacacagttgttctttcagaaaatattccactcaacaccaaaattgctctc 1976 V. 92aacggagaagaggacaatgagtaagctaaact25 V.8 1992 ataactgtgacggataaggatgcggaccataatggcagggtgacatgcttcacagatcat 2051 V.1 2052 gaaatccctttcagattaaggccagtattcagtaatcagttcctcctggagactgcagca 2111 V.8 2037 gaaattcctttcagattaaggccagtattcagtaatcagttcctcctggagaatgcagca 2096 V.1 2112 tatcttgactatgagtccacaaaagaatatgccattaaattactggctgcagatgctggc 2171 2097 tatcttgactatgagtccacaaaagaatatgccattaaattactggctgcagatgctggc 2156 00V 1 2 7 a c t c t g at a t a c a g t t c t a ag g a g t a a t a a t 2 3 00V.1 217 aaacctcctttgaatcagtcagcaatgctcttcatcaaagtgaaagatgaaaatgacaat 2216 V.1 2232 gctccagttttcacccagtctttcgtaactgtttctattcctgagaataactctcctggc 2291 V.8 2217 gctccagttttcacccagtctttcgtaactgtttctattcctgagaataactctcctggc 2276 V.1 2292 atccagttgacgaaagtaagtgcaatggatgcagacagtgggcctaatgctaagatcaat 2351 V.8 2277 atccagttgatgaaagtaagtgcaacggatgcagacagtgggcctaatgctgagatcaat 2336 V.1 *2352 tacctgctaggccctgatgctccacctgaattcagcctggattgtcgtacaggcatgctg 2411 V.8 :2337 tacctgctaggccctgatgctccacctgaattcagcctggatcgtcgtacaggcatgctg 2396 V.1 :2412 actgtagtgaagaaactagatagagaaaaagaggataaatatttattcacaattctggca 2471 V.8 :2397 actgtagtgaagaaactagatagagaaaaagaggataaatatttattcacaattctggca 2456 V.1 :2472 aaagataacggggtaccacccttaaccagcaatgtcacagtctttgtaagcattattgat 2531 V.8 :2457 aaagataatggggtaccacccttaaccagcaatgtcacagtctttgtaagcattattgat 2516 V.1 :2532 cagaatgacaatagcccagttttcactcacaatgaatacaacttctatgtcccagaaaac 2591 V.8 :2517 cagaatgacaatagcccagttttcactcacaatgaatacaaattctatgtcccagaaaac 2576 V.1 :2592 cttccaaggcatggtacagtaggactaatcactgtaactgatcctgattatggagacaat 2651 V.8 :2577 cttccaaggcatggtacagtaggactaatcactgtaactgatcctgattatggagacaat 2636 V.1 :2652 tctgcagttacgctctccattttagatgagaatgatgacttcaccattgattcacaaact 2711 V.8 2637 tctgcagttacgctctccattttagatgagaatgatgacttcaccattgattcacaaact 2696 V.1 2712 ggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatcttacactttctat 2771 V.8 2697 ggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatcttacactttctat 2756 293 00 V.1 2772 gtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaagtaaccataaat 2831 V.8 2757 gtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaagtaaccataaat 2816 V.1 2832 gtggttgatgtcaatgacaacaaaccagttttcattgtccctccttccaactgttcttat 2891 11111 ll lllIIItiilI~ilill~ II Il~ll Hill11 1111111 V.8 2817 gtggttgatgtcaatgacaacaaaccagttttcattgtccctccttacaactattcttat 2876 V.1 2892 gaattggttctaccgtccactaatccaggcacagtggtctttcaggtaattgctgttgac 2951 V.8 2877 gaattggttctaccgtccactaatccaggcacagtggtctttcaggtaattgctgttgac 2936 V. 92ataatgctatcggtcgtcgattgagaaaaggt31 V.81 2952 aatgacactggcatgaatgcagaggttcgttacagcattgtaggaggaaacacaagagat 3011 00 V.1 :3012 ctgtttgcaatcgaccaagaaacaggcaacataacattgatggagaaatgtgatgttaca 3071 V.8 :2997 ctgtttgcaatcgaccaagaaacaggcaacataacattgatggagaaatgtgatgttaca 3056 V.1 :3072 gaccttggtttacacagagtgttggtcaaagctaatgacttaggacagcctgattctctc 3131 V.8 :3057 gaccttggtttacacagagtgttggtcaaagctaatgacttaggacagcctgattctctc 3116 V.1: 3132 ttcagtgttgtaattgtcaatctgttcgtgaatgagtcggtgaccaatgctacactgatt 3191 V.8: 3117 ttcagtgttgtaattgtcaatctgttcgtgaatgagtcagtgaccaatgctacactgatt 3176 V.1 :3192 aatgaactggtgcgcaaaagcactgaagcaccagtgaccccaaatactgagatagctgat 3251 V.8 :3177 aatgaactggtgcgcaaaagcattgaagcaccagtgaccccaaatactgagatagctgat 3236 V.1: 3252 gtatcctcaccaactagtgactatgtcaagatcctggttgcagctgttgctggcaccata 3311 V.8 3237 gtatcctcaccaactagtgactatgtcaagatcctggttgcagctgttgctggcaccata 3296 V.1 3312 actgtcgttgtagttattttcatcactgctgtagtaagatgtcgccaggcaccacacctt 3371 V.8 3297 actgtcgttgtagttattttcatcactgctgtagtaagatgtcgccaggcaccacacctt 3356 V.1 3372 aaggctgctcagaaaaacaagcagaattctgaatgggctaccccaaacccagaaaacagg 3431 V.8 3357 aaggctgctcagaaaaacatgcagaattctgaatgggctaccccaaacccagaaa~cagg 3416 V.1 :3432 cagatgataatgatgaagaaaaagaaaaagaagaagaagcattcccctaagaacttgctg 3491 V.8 :3417 cagatgataatgatgaagaaaaagaaaaagaagaagaagcattcccctaagaacctgctg 3476 V.1: 3492 cttaattttgtcactattgaagaaactaaggcagatgatgttgacagtgatggaaacaga 3551 V.8 3477 cttaatgttgtcactattgaagaaactaaggcagatgatgttgacagtgatggaaacaga 3536 V.1 3552 gtcacactagaccttcctattgatctagaagagcaaacaatgggaaagtacaattgggta 3611 V.8 3537 gtcacactagaccttcctattgatctagaagagcaaacaatgggaaagjtacaattgggta 3596 00 V.1 :3612 actacacctactactttcaagcccgacagccctgatttggcccgacactacaaatctgcc 3671 V.8 :3597 actacacctactactttcaagcctgacagccctgatttggcccgacactacaaatctgcc 3656 V.1 :3672 tctccacagcctgccttccaaattcagcctgaaactcccctgaattcgaagcaccacatc 3731 v.8 3657 tctccacagcctgccttccaaattcagcctgaaactcccctgaatttgaagcaccacatc 3716 V.1 3732 atccaagaactgcctctcgataacacctttgtggcctgtgactctatctccaagtgttcc 3791 V.8 3717 atccaagaactgcctctcgataacacctttgtggcctgtgactctatctccaattgttcc 3776 V. 72tagattaaccaacttcgcggcacatgcactgg35 3792 tcaagcagttcagatccctacagcgtttctgactgtggctatccagtgacgaccttcgag 3851 00 V.1 3852 gtacctgtgtccgtacacaccagaccg 3878 V.8 3837 gtacctgtgtccgtacacaccagaccg 3863 Score 1567 bits (815), Expect -O.Oldentities 829/836 Strand PlusI Plus V.1 3 ggtggtccagtacctccaaagatatggaatacactcctgaaatatcctgaaaactttttt 62 V.8 1 ggtggtccagtacctccaaagatatggaatacactcctgaaatatcctgaaacctttttt V.1 63 ttttcagaatcctttaataagcagttatgtcaatctgaaagttgcttacttgtactttat 122 V. 8 61 ttttcagaatcctttaataagcagttatgtcaatctgaaagttgcttacttgtactttat 120 V.1 123 attaatagctattcttgtttttcttatccaaagaaaaatcctctaatccccttttcacat 182 V.8 121 attaatagctattcttgtttttcttatccaaagaaaaatcctctaatccccttttcacat 180 V.1 183 gatagttgttaccatgtttaggcattagtcacatcaacccctctcctctcccaaacttct 242 V.8 181 gatagttgttaccatgtttaggcgttagtcacatcaacccctctcctctcccaaacttct 240 V.1 243 cttcttcaaatcaaactttattagtccctcctttataatgattccttgcctcgttttatc 302 V.8 241 cttcttcaaatcaaactttattagtccctcctttataatgattccttgcctccttttatc 300 V. 1 303 cagatcaattttttttcactttgatgcccagagctgaagaaatggactactgtataaatt 362 V.8 301 cagatcaattttttttcactttgatgcccagagctgaagaaatggactattgtataaatt 360 V.1 363 attcattgccaagagaataattgcattttaaacccatattataacaaagaataatgatta 422 V.8 :361 attcattgccaagagaataattgcattttaaacccatgttataacaaagaataatgatta 420 V.1 :423 tattttgtgatttgtaacaaataccctttattttcccttaactattgaattaaatatttt 482 V. B :421 tattttgtgatttgtaacaaataccctttattttcccttaactattgaattaaatatttt 480 V.1 483 aattatttgtattctctttaactatcttggtatattaaagtattatcttttatatattta 542 V.8 481 aattatttgtattctctttaactatcttggtatattaaagtattatcttttatatattta 540 V.1 543 tcaatggtggacacttttataggtactctgtgtcatttttgatactgtaggtatcttatt 602 V.8 541 tcaatggtggacacttttataggtactctgtgtcatttttgatactgtaggtatcttatt 600 V.1 603 tcatttatctttattcttaatgtacgaattcataatatttgattcagaacaaatttatca 662 V.8 601 tcatttatctttattcttaatgtacgaattcataatatttgattcagaacagatttatca 660 V.1 663 ctaattaacagagtgtcaattatgctaacatctcatttactgattttaatttaaaacagt 722 V.8 661 ctaattaacagagtgtcaattatgctaacatctcatttactgattttaatttaaaacagt 720 V.1 :723 ttttgttaacatgcatgtttagggttgcttcttaataatttcttcttcctcttctctct 782 V.8 :721 ttttgttaacatgcatgtttagggttggcttcttaataatttcttcttcctcttctctct 780 V.1: 783 ctcctcttcttttggtcagtgttgtgcgggttaatacaacaaactgtaacaagtgt 838 V.8: 781 ctcctcttcttttggtcagtgttgtgcgggttaatacaacaaactgtcacaagtgt 836 Table LIV(g). Peptide sequences of protein coded by 109PID4 v.8 (SEQ ID NO: 276) MFRVGFLI IS

TIREEIPENV

ARI DREKLCA

PENSAINSKY

REEKDTYVM(

TQLHATDADI

ASDGGLMPAR

DADHNGRVTC

SAMILFIKVKD

APPEFSLDRR

VFTHNEYKFY

ISFDREKQES

TI4PGTVVFQV

VLVKANDLGQ

DYVKILVAAV

KKKKKKKHSP

KPDS PDLARH

YSVSDCGYPV

PLGYPQEEYF

HSDACWMPAS

CHSPPPIQVS

MRS QAQS SVS

PRQQARPSRG

SSSSLSPLLL

TLIGNLLKDLN

GIPRDEHCFY

TLPAAVDPDV

VKVEDGGFPQ

GENAKIHFSF

AMVLVNVTDV

FTDHEI PFRL

ENDNAPVFTQ

TGMLTVVKKL

VPENLPRHGT

YTFYVKAEDG

IAVDNDTGMN

PDSLFSVVIV

AGTITVVVVI

KNLLLNVVT I

YKSASPQPAF

TTFEVPVSVH

DRATPSNRTE

LDHSSSSQAQ

ALHHS PPLVQ

LQQGWVQGAN

DSPIMETHPL

VSVVRVNTTN

IJSLIPNKSLT

EVEVAILPDE

GINGVQNYEL

RSSTAILQVS

SNLVSNIARR

NDNVPSIDIR

RPVFSNQFLL

SFVTVSIPEN

DREKEDKYLF

VGLITVTDPD

GRVSRSSSAK

AEVRYSIVGG

NLFVNESVTN

FITAVVRCRQ

EETKADDVDS

QIQPETPLNL

TRPSQRRVTF

GDGNSDPEST

ASAIJCHS PPL GTALHHS PPS

GLCSVDQGVQ

CHKCLLSGTY

TTMQFKLVYK

I FRLVKIRFL IKSQNI FGLD

VTDTNDNHPV

LFHLNATTGL

YIVNPVNDTV

ENAAYLDYES

NSPGIQLMKV

TILAKDNGVP

YGDNSAVTLS

VTINVVDVND

NTRDLFAIDQ

ATLINELVRK

APHLKAAQKN

DGNRVTL DL P KHHI11IQELPL

HLPEGSQESS

FIPGLKKEIT

SQASTQHHSP

AQASALCYS P

GSATSQFYTM

I FAVLLVCVV

TGDVPLIRIE

IEDINDNAPL

VIETPEGDKM

FKETEIEVSI

ITIKEPLDRE

VISENIPLNT

TKEYAIKLLA

SATDADSGPN

PLTSNVTVFV

ILDENDDFTI

NKPVFIVPPY

ETGNITLMEK

SIEAPVTPNT

MQNSEWATPN

I DLEEQTMGK

DNTFVACDSI

SDGGLGDHDA

VQPTVEEASD

PVTQTIVLCH

PLAQAAAISH

SERLHPSDDS

FHSGAQEKNY

EDTGEI FTTG

FPATVINISI

PQLIVQKELD

PENAPVGTSV

ETPNHKLLVL

KIALITVTDK

ADAGKPPLNQ

AEINYLLGPD

SIIDQNDNSP

DSQTGVIRPN

NYSYELVLPS

CDVTDLGLHR

EIADVSSPTS

PENRQMIMMK

YNWV'rTPTTF

SNCSSSSSDP

GSLTSTSHGL

NCTQECLIYG

SPPVTQTIAL

SSSLPQVIAL

IKVI PLTTFA 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1340 Table LV(g). Amino acid sequence alignment of 109P1 D4 v.1 (SEQ ID NO: 277) and 109P1D4 v.8 (SEQ ID NO: 278) Score 1961 bits (5081), Expect =0.Oldenfites 99211009 Posives 99511009 (98%) V.1 :3 LLSGTYIFAVLLACVVFHSGAQEKNYTIREE4PENVLIGDLLKDLNLSLIPNKSLTTAMQ 62 LLSGTYIFAVLL CVVFHSGAQEKNYTIREE+PENVLIG+LLKDLNLSLIPNKSLTT MQ V.8 :35 LLSGTYIFAVLLVCVVFHSGAQEKN'YTIREIPENVLIGNLLKDLI;LSLIPNKSLTTTMQ 94 V.1 63 FKLVYKTGDVPLIRIEEDTGEIrrTGA1RIDREKLCAGIPRDEHCFYEVEVAILPDEIFRL 122

FKLVYKTGDVPLIRIEEDTGEIFTTGARIDREKLCAGIPRDEHCFYEVEVAILPDEIFRL

V.8 95 FtVYKTGDVPLIRIEEDTGEINTTGARIDREKCAGIPRlEHCFEVEVATLPDETrRL 154 00 oV.1 123 VKIRFLIEDINDNAPLFPATVINISIPENsAINSKYTLpAAVDPDVGINGVQNYELIKSQ 182 oKRLEIDALPTIIIESANKTPADDGNVNEIS V.8 155 VKIRFLIEDINDNAPLFPATVINISIPENSAINsKYTLPAAVDPDVGINGVQNYELIKSQ 214 V.1 183 NIFGLDVIETPEGDKMPQLIVQKELDREEKDTYVMKVKVEDGGFPQRSSTAILQVSVTDT 242 NIFG3LDVIETPEGDKMPQLIVQKELDREEKDTYVMKVKVEDGGFPQRSSTAILQVSVTDT CAv.8 215 NIFGLDVIETPEGDKMPQLIVQKELDREEKDTYVtKVKVEDGGFPQRSSTAILQVSVTDT 274 V.1 243 NDNHPVFKETEIEVSIPENAPVGTSVTQLHATDADIGENAKIHFSFSNLVSNIARRLFHL 302 0 NDNHPVFK<ETEIEVSIPENAPVGTSVTQLHATDADIGENAKIHFSFSNLVSNIARRLFHL V.8 :275 NDNHPVFKETEIEVSIPENAPVGTSVTQLHATDADIGENAKIHFSFSNLVSNIARRLFHL 334 OV.1 :303 NATTGLITIKEPLDREETPNHKLLVLASDGGLMPARAMVLVNVTDVNDNVPSIDIRYIVN 362 cINATTGLITIKEPLDREETPNHKLLVLASDGLPARAVLVNVTDVNDNVPSIDIYV oV.8 :335 NATTGLITIKEPLDREETPNHKLLVLASDGGIMPARAMVLVNVTDVNDNVPSIDIRYIVN 394 V. 6 VDVcEILTILTTKADNRTiDEPRRVSQLEA 2 V.1 363PNDTVVLSENIPLNTKIALITVTDKDADHNGRVTCFTDHEIPFRLRPVFSNOFLLETAA42 00 V.8 395PVNDTWISENI 00V.8 :43 PVDETVLENAIPLNTDKIALDKADHNGRVTCFTDI{EPVFRLRVSQFLENAA 454 0 V.1 423 YLDYESTKEYAIKLLAADAGKPPI.NQSANLFIKVKDENDNAPVFlQSFVTVSIPENNSPG48 V.0 5 YLDYESTKEYAIKLLAADAGKPPLNQSAMLFIKVKDENDNAPVFTQSFVTVSIPENNSPG51 V.1 :483 IOLTKVSAMDADSGPNAKINYLLGPDAPPEFSLDCRTGMLTVVKICLDREKEDKYLFlTILA 542 IQL KVSA DADSGPNA+INYLLGPDAPPEFSLD RTGMLTVVKKLDREKEDKYLE7ILA V.8 :515 IQLMKVSATDADSGPNAEINYLLCPDAPPEFSLDRRTGMIJTVVKKLDREKEDKYLFIILA 574 V.1 :543 KDNGVPPLTSNVTVFVSIIDQNDNSPVFTHNEYNFYVPENLPRHGTVGLITVTDPDYGDN 602 KDNGVPPLTSNVTVFVSI IDQNDNSPVFT-NEY FYVPENLPRHGTVGLITVTDPDYGDN V.8 :575 KDNGVPPLTSNV'rVFVSI IDQNDNSPVFTHNEYKFYVPENLPRHGTVGLITVTDPDYGDN 634 V.1: 603 SAVTLSILDENDDFTIDSQTGVIRPNISFDREKQESYTFYVKAEDGGRVSRSSSAKVTIN 662

SAVTLSILDENDDFTIDSQTGVIRPNISFDREKQESYTFYVCAEDGGRVSRSSSAXVTIN

V.8: 635 SAVTLSILDENDDFTIDSQTGVIRPNISFDREKQESYTFYVCAEDGGRVSRSSSAKVTIN 694 V.1 :663 VVDVNDNKPVFIVPPSNCSYELVLPSTNPGTVVFQVIAVDNDTGMNAEVRYSIVGGNTRD 722 VVDVNDNKPVFIVPP N S.YELVLPSTNPGTWFQVIAVDNDTGMNAEVRYSIVGGNTRD V.8 :695 VVDVNDNKPVFIVPPYNYSYELVLPSTNPGTVVFVIAVDNDTG4NAEVRYSIVGGNTRD 754 V.1 723 LFAIDQETGNITL4EKCDVTDLGLHRVLVKANDLGQPDSLFSVVIVNLFVNESVTNATLI 782 LFAI DQETGNITLMElCCDVTDLGLHRVLVXANDLGQPDSLFSVVIVNLFVNESVTNATLI V.8 755 LFAIDQETGNITLNEKCDVTDLGLHRVLVKANDLGQPDSLFSVVIVNLFVNESVTNATLI 814 V.1 783 NELVRXSTEAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVVVIFITAVVRCRQAPHL 842 NELVRCS EAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVVVIFITAVVRCRQAPHL V.8 :815 NELVRKSIEAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVVVIFITAVVRCROAPHL 874 V.1 :843 KAAQKNKQNSEWATPNPENRQMIMI4KKKKKKKKHSPKNLLLNFVTIEETKADDVDSDGNR 902 KAAQKN QNSEWATPNPENRQMIHMKKKKKKKKHSPKNLLLN VTIEETKADDVDSDGNR V.8 :875 KAAQKNMQNSEWATPNPENRQMIMMKKKKKKKKHSPIQUJLLNVVTIEETKADDVDSDGNR 934 V.1 :903 VTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLARHYKSASPQPAFQIQPETPLNSKHHI 962 VTLDLPIDLEEQTMGKYNWVTTPTTFKPDSPDLARHYKSASPQPAFQIQPETPLN KHHI V.8 :935 VTLDLPIDLEEOTMGICYNWVTTPTTFKPDSPDLARHYKSASPQPAFQIQPETPLNLKHHI 994 V.1 :963 IOELPLDNTFVACDSISKCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP 1011 IQELPLDNTFVACDSIS CSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP V.8 :995 IQELPLDNTFVACDSISNCSSSSSDPYSVSDCGYPVTTFEVPVSVIiTRP 1043 Table UI(h). Nucleotide sequence of transcript variant 109PI D4S v.9 (SEQ ID NO: 279) cccctttctc cccctctgtt aagtccctcc ccctcgccat tcaaaagggc tggctcggca ctggctcctt gcagtcggcg aactgtctgg gcgggaggag ccgtgagcag tagctgcact 120 cagctgcccg cgcggcaaag aggaaggcaa qccaaacaga gtgcgcagag tggcagtgcc 180 agcggcgaca caggcagcac aggcagcccg ggctgcctga atagcctcag aaacaacctc 240 agcgactccg gctgctctgc ggactgcgag ctgtggcggt agagcccgct acagcagtcg 300 cagtctccgt ggagcgggcg gaagcctttt ttctcccttt cgtttacctc ttcattctac 360 00 00 tctaaaqgca taccagaqg gcttggctga tcaagtgttg attttcgcgg accatccgag ttgtcgctga accggagatg gctcgcattg gaagtggagg atagaagata ccagagaact ggcataaacg gtcattgaaa agggaagaga agatccagta tttaaggaga acacagctcc agcaatctag atcacaatca gcaagtgatg aatgataatg gttctttcag gatgcggac aggccagtat acaaaagaat tcagcaatqc tctttcgtaa agtgcaacgg gctccacctg gatagagaaa cccttaacca gttttcactc gtaggactaa attttagatg atttcatttg ggtagagtat aacaaaccag actaatccag gcagaggttc gaaacaggca gtgttggtca aatctgttcg agcattgaag gactatgtca ttcatcactg atgcagaatt aaaaagaaaa gaagaaacta attgatctag aagcctgaca caaattcagc gataacacct tacagcgttt accagaccga aggaacaaca ggctaagatc ttaaaaatga ttaaaattta gtttgctctg gttaatatat aacattatct tcgttattag gttttqcctc ttgcagtgca tgcgggttaa tcctgctagt aagaaattcc ttccaaacaa tgccactgat atcgtgagaa ttgccatttt taaatgataa cggctataaa qagttcaaaa caccagaagg aggataccta ctgctatttt cagagattga atgccacaga tctccaacat aagaaccact gtggattgat tcccatccat aaaatattcc ataatggcag tcaqtaatca atgccattaa tcttcatcaa ctgtttctat atgcagacag aattcagcct aagaggataa gcaatgtcac acaatgaata tcactgtaac agaatgatga atagagaaaa cacgttcttc ttttcattgt gcacagtggt gttacagcat acataacatt aagctaatga tgaatgagtc caccagtgac agatcctggt ctgtagtaag ctgaatgggc agaagaagaa aggcagatga aagagcaaac gccctgattt ctgaaactcc ttgtggcctg ctgactgtgg ctgattccag aaattccatt attaattttg tgtcctagtg tggaagagac aatcaacagc cagtcatgaa ccaggagttt gaaaatcctq agctgctctc ctatgaggac tacaacaaac atgcgtggtg agaaaacgtc gtccttgaca tcgaattgaa attatgtgct gccggatgaa tgcaccattg ctctaaatat ctacgaacta agacaagatg tgtgatgaaa gcaagtaagt agtcagtata tgctgacata tgccaqgaqa ggatagggaa qccagcaaga tgacataaga actcaacacc ggtgacatgc gttcctcctg attactggct agtgaaagat tcctgagaat tgggcctaat gqatcgtcgt atatttattc agtctttgta caaattctat tgatcctgat cttcaccatt acaagaatct aagtgccaaa ccctccttac ctttcaggta tgtaggagga gatggagaaa cttaggacag agtgaccaat cccaaatact tgcagctgtt atgtcgccag taccccaaac gcattcccct tgttgacagt aatgggaaag ggcccgacac cctgaatttg tgactctatc ctatccagtg gacatqaact ccccttccaa taatctagat aaccttgtgc aqtgcagcgc catgatgtaa acatgcaatt ggaagtgagc ttqtgaataa aactttgtaa tgaatgacag tgtcacaagt ttccactctg ct gataggca actactatgc gaggatactg ggtatcccaa atatttagac ttcccagcaa actctcccag attaagagtc ccacaactga gtaaaggttg gttactgata ccagaaaatg ggtgaaaatg ttatttcacc gaaacaccaa gcaatggtgc tacatcgtca aaaattgctc ttcacagatc gagaatgcag gcagatgctg gaaaatgaca aactctcctg gctgagatca acaggcatgc acaattctgg agcattattg gtcccagaaa tatggagaca gattcacaaa tacactttct gtaaccataa aactattctt attgctgttg aacacaagag tgtgatgtta cctgattctc gctacactga gagatagctg gctggcacca gcaccacacc ccagaaaaca aagaacctgc gatggaaaca tacaattggg tacaaatctg aagcaccaca tccaattgtt acaaccttcg attgaaatct aaaatttcaa ttcccattat tttctttagc aataacagag tataaggctg acttgccctg tgaactagcc gaaggattcc tcttqtgaag tgggttttaa gtttgttgtc gcgcccagga acttgttgaa agttcaaqct gtgagatctt gggatgagca tggttaagat cagttatcaa cggctgttga aaaacatttt ttgttcaaaa aagatggtgg caaatgacaa ctcctgtagg ccaagatcca tcaatgccac accacaagtt tggtaaatgt atcctgtcaa tcataactgt atgaaattcc catatcttga gcaaacctcc atgctccagt gcatccagtt attacctgct tgactgtagt caaaagataa atcagaatga accttccaag attctqcagt ctggtgtcat atgtaaaggc atgtggttga atgaattggt acaatgacac atctgtttgc cagaccttgg tcttcagtgt ttaatgaact atgtatcctc taactgtcgt ttaaggctgc ggcagatgat tgcttaatgt gagtcacact taactacacc cctctccaca tcatccaaga cctcaagcag aggtacctgt gcagtgagat tgattgtgat aaaagcaagc tgtaatctgg tactctcatg tcttggtgta tctgattgtt aaactactct acagatcaca aagctgacaa ttcagatatt cgggacgtac gaaaaactac agaccttaac agtgtacaag cactaccggc ttgcttttat acgttttctg catatcaatt tcctgacgta tggcctcgat qgagttagat ctttcctcaa ccacccagtc cacttcagtg cttctctttc cactggactt actggttttg tacagatgtc tgacacagtt gacggataag tttcagatta ctatgagtcc tttgaatcag tttcacccag gatqaaaqta aggccctgat gaagaaacta tggggtacca caatagccca gcatggtaca tacgctctcc ccgaccaaat tgaggatggt tgtcaatgac tctaccgtcc tggcatgaat aatcgaccaa tttacacaga tgtaattgtc ggtgcgcaaa accaactagt tgtagttatt tcagaaaaac aatgatgaag tgtcactatt agaccttcct tactactttc gcctgccttc actgcctctc ttcagatccc gtccgtacac gtaactttct ttcaaaatta aaaaatcatc caatggaaat ctgtttctct tacacttatg gaataattaa ctgaaaggta 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 298 00 tccagggcaa gagacatttt taagacccca aacaaacaaa aaacaaaacc aaaacactct 4140 ggttcagtgt tttgaaaata ttgactaaca taatattgct gagaaaatca tttttattac 4200 ccaccactct gcttaaaagt tgagtgggcc gggcgcggtg gctcacgcct gtaattccag 4260 cactttggga ggccgaggcg ggtggatcac qagqtcagga tattgagacc atcctggcta 4320 acatggtgaa accccatctc cactaaaaat acaaaaaatt agctgggcgt ggtggcgggc 4380 gcctgtagtc ccagctactc gggaggctga ggcaggagaa tggcgtgaac ccgggaggcg 4440 gagcttgcag tgagccgaga tggcgccact gcactccaqc ctgggtgaca gagcaagact 4500 ctgtctcaaa aagaaaaaaa tgttcagtga tagaaaataa ttttactagg tttttatgtt 4560 gattgtactc atgctgttcc actcctttta attattaaaa agttattttt ggctgggtgt 4620 ggtggctcat acctgtaatc ccagcacttt gggaggccga ggcgggtgga tcacctgagg 4680 tcaggagttc aagaccagtc tggccaacat 4710 Table Ull(h). Nucleotide sequence alignment of 109PI D4 v.1 (SEQ ID NO: 280) and 109P1 D4 v.9 (SEQ ID NO: 281) ___Score =5664 bits (2946), Expect 0.Oldentities 3000/3027 Strand Plus Plus V.1 852 ttgttgtccgggacgtacattttcgcggtcctgctagcatgcgtggtgttccactctggc 911 00 111 1111 I 111 1111111111111 ii11 V.9 583 ttgttgtccgggacgtacattttcgcggtcctgctagtatgcgtggtgttccactctggc 642 V.1 912 gcccaggagaaaaactacaccatccgagaagaaatgccagaaaacgtcctgataggcgac 971 V.9 643 gcccaggagaaaaactacaccatccgagaagaaattccagaaaacgtcctgataggcac 702 V.1 972 ttgttgaaagaccttaacttgtcgctgattccaaacaagtccttgacaactgctatgcag 1031 V.9 703 ttgttgaaagaccttaacttgtcgctgattccaaacaagtccttgacaactactatgcag 762 V.1 1032 ttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaagaggatactggt 1091 V.9 763 ttcaagctagtgtacaagaccggagatgtgccactgattcgaattgaagaggatactggt 822 V.1 1092 gagatcttcactactggcgctcgcattgatcgtgagaaattatgtgctggtatcccaagg 1151 V.9 823 gagatcttcactaccggcgctcgcattgatcgtgagaaattatgtgctggtatcccaagg 882 V.1 1152 gatgagcattgcttttatgaagtggaggttgccattttgccggatgaaatatttagactg 1211 V. 9 883 gatgagcattgcttttatgaagtggaggttgccattttgccggatgaaatatttagactg 942 V.1 1212 gttaagatacgttttctgatagaagatataaatgataatgcacc-attgttcccagcaaca 1271 V.9 943 gttaagatacgttttctgatagaagatataaatgataatgcaccattgttcccagcaaca 1002 V.1 1272 gttatcaacatatcaattccagagaactcggctataaactctaaatatactctcccagcg 1331 V.9 1003 gttatcaacatatcaattccagagaactcggctataaactctaaatatactctcccagcg 1062 V.1 1332 gctgttgatcctgacgtaggaataaacggagttcaaaactacgaactaattaagagtcaa 1391 V.9 1063 gctgttgatcctgacgtaggcataaacggagttcaaaactacgaactaattaagagtcaa 1122 V.1 1392 aacatttttggcctcgatgtcattgaaacaccagaaggagacaagatgccacaactgatt 1451 V.9 1123 aacatttttggcctcgatgtcattgaaacaccagaaggagacaagatgcacaactgatt 1182 V.1 1452 gttcaaaaggagttagatagggaagagaaggatacctacgtgatgaaagtaaaggttgaa 1511 299 00V.9 1183 gttcaaaaggagttagatagggaagagaaggatacctatgtgatgaaagtaaaggttgaa 1242 V.1 1512 gatggtggctttcctcaaagatccagtactgctattttgcaagtgagtgttactgataca 1571 V.9 1243 gatggtggctttcctcaaagatccagtactgctattttgcaagtaagtgttactgataca 1302 V.1 1572 aatgacaaccacccagtctttaaggagacagagattgaagtcagtataccagaaaatgct 1631 V. 9 1303 aatgacaaccacccagtctttaaggagacagagattgaagtcagtataccagaaaatgct 1362 C1V.1 1632 cctgtaggcacttcagtgacacagctccatgccacagatgctgacataggtgaaaatgcc 1691 (N VA 1363 cctgtaggcacttcagtgacacagctccatgccacagatgctgacataggtgaaaatgcc 1422 00 V.1 1692 aagatccacttctctttcagcaatctagtctccaacattgccaggagattatttcacctc 1751 V.9 1423 aagatccacttctctttcagcaatctagtctccaacattgccaggagattatttcacctc 1482 V. 1 1752 aatgccaccactggacttatcacaatcaaagaaccactggatagggaagaaacaccaaac 1811 V.9 1483 aatgccaccactggacttatcacaatcaaagaaccactggatagggaagaaacaccaaac 1542 V.1 1812 cacaagttactggttttggcaagtgatggtggattgatgccagcaagagcaatggtgctg 1871 V. 9 1543 cacaagttactggttttggcaagtgatggtggattgatgccagcaagagcaatggtgctg 1602 V.1 :1872 gtaaatgttacagatgtcaatgataatgtcccatccattgacataagatacatcgtcaat 1931 V.9 :1603 gtaaatgttacagatgtcaatgataatgtcccatccattgacataagatacatcgtcaat 1662 V.1 :1932 cctgtcaatgacacagttgttctttcagaaaatattccactcaacaccaaaattgctctc 1991 V.9 9 1663 cctgtcaatgacacagttgttctttcagaaaatattccactcaacaccaaaattgctctc 1722 V. 1 :1992 ataactgtgacggataaggatgcggaccataatggcagggtgacatgcttcacagatcat 2051 V.9 :1723 ataactgtgacggataaggatgcggaccataatggeagggtgacatgcttcacagatcat 1782 V.1 :2052 gaaatccctttcagattaaggccagtattcagtaatcagttcctcctggagactgcagca 2111 Hill111111 11~~l 111111111111 1111111 VA. 1783 gaaattcctttcagattaaggccagtattcagtaatcagttcctcctggagaatgcagca 1842 V.1 :2112 ta~ttgactatgagtccacaaaagaatatgccattaaattactggctgcagatgctggc 2171 V.9 :1843 tatcttgactatgagtccacaaaagaatatgccattaaattactggctgcagatgctggc 1902 V.1 :2172 aaacctcctttgaatcagtcagcaatgctcttcatcaaagtgaaagatgaaaatgacaat 2231 V.9 :1903 aaacctcctttgaatcagtcagcaatgctcttcatcaaagtgaaagatgaaaatgacaat 1962 V.1 :2232 gctccagttttcacccagtctttcgtaactgtttctattcctgagaataactctcctggc 2291 V.9 :1963 gctccagttttcacccagtctttcgtaactgtttctattcctgagaataactctcctggc 2022 V.1 :2292 atccagttgacgaaagtaagtgcaatggatgcagacagtgggcctaatgctaagatcaat 2351 300 0 011 1 11 1 111 1 1 1 v.9 2023 atccagttgatgaaagtaagtgcaacggatgcagacagtgggcctaatgctgagatcaat 2082 V. 32tctcagccgtccacgatcgcgattgtcgctcg21 v.9 203 tacctgctaggccctgatgctccacctgaattcagcctggattgtcgtacaggcatgctg 241 V.1 2412 actgtagtgaagaaactagatagagaaaaagaggataaatatttattcacaattctggca 2471 V.9 2143 8 ctgtagtgaagaaactagatagagaaaaagaggataaatatttattcacaattctggca 2202 V.1 2472 aaagataacggggtaccacccttaaccagcaatgtcacagtctttgtaagcattattgat 2531 V.9 2203 aaagataatggggtaccacccttaaccagcaatgtcacagtctttgtaagcattattgat 2262 00 V.1 :2532 cagaatgacaatagcccagttttcactcacaatgaatacaacttctatgtcccagaaaac 2591 V.9 :2263 cagaatgacaatagcccagttttcactcaCaatgaatacaaattctatgtcccagaaaac 2322 V.1 :2592 ctcagctgaatgatacattatactattggca 2651 V.9 :2323 ctcagctgaatgatacattatactattggca 2382 V.1 :2652 tctgcagttacgctctccattttagatgagaatgatgacttcaccattgattcacaaact 2711 V.9 :2383 tctgcagttacgctctccattttagatgagaatgatgacttcaccattgattcacaaact 2442 V.1 :2712 ggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatcttacactttctat 2771 V.9 2443 ggtgtcatccgaccaaatatttcatttgatagagaaaaacaagaatcttacactttctat 2502 V.1 2772 gtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaagtaaccataaat 2831 V. 9 2503 gtaaaggctgaggatggtggtagagtatcacgttcttcaagtgccaaagtaaccataaat 2562 V.1 2832 gtggttgatgtcaatgacaacaaaccagttttcattgtccctccttccaactgttcttat 2891 V.9 2563 gtggttgatgtcaatgacaacaaaccagttttcattgtccctccttacaactattcttat 2622 V.1 2892 gaattggttctaccgtccactaatccaggcacagtggtctttcaggtaattgctgttgac 2951 V. 9 2623 gaattggttctaccgtccactaatccaggcacagtggtctttcaggtaattgctgttgac 2682 V.1 2952 aatgacactggcatgaatgcagaggttcgttacagcattgtaggaggaaacacaagagat 3011 V.9 2683 aatgacactggcatgaatgcagaggttcgttacagcattgtaggaggaaacacaagagat 2742 V. 1 3012 ctgtttgcaatcgaccaagaaacaggcaacataacattgatggagaaatgtgatgttaca 3071 V.9 2743 ctttcacacaaaagacaactgtggattagtc 2802 V.1 3072 gaccttggtttacacagagtgttggtcaaagctaatgacttaggacagcctgattctctc 3131 V.9 2803 gaccttggtttacacagagtgttggtcaaagctaatgacttaggacagcctgattctctc 2862 301 00 v.1 3132 ttcagtgttgtaattgtcaatctgttcgtgaatgagtcggtgaccaatgctacactgatt 3191 (N v.9 2863 ttcagtgttgtaattgtcaatctgttcgtgaatgagtcagtgaccaatgctacactgatt 2922 V.1 3192 aatgaactggtgcgcaaaagcactgaagcaccagtgaccccaaatactgagatagctgat 3251 V.9 2923 aatgaactggtgcgcaaaagcattgaagcaccagtgaccccaaatactgagatagctgat 2982 V.1 3252 gtatcctcaccaactagtgactatgtcaagacctggttgcagctgttgctgcaccata 3311 V.9 2983 gtatcctcaccaactagtgactatgtcaagatcctggttgcagctgttgctggcaccata 3042 V. 32attgtgatattaccgcgataagccagccaact37 (i V.1 303 actgtcgttgtagttattttcatcactgctgtagtaagatgtcgccaggcaccacacctt 331 00 V.1: 3372 aaggctgctcagaaaaacaagcagaattctgaatgggctaccccaaacccagaaaacagg 3431 (i V.9 :3103 aagtccgaactcgattgaggtccaacaaacg 3162 V.1 :3432 caagtagtagaagaagagaagatccagatgt 3491 V. 9 :3163 caagtagtagaagaagagaagatccagacgt 3222 V.1 :3492 ctattgccatagacaggaagttgcggtgacg 3551 HIM1liI1i1l 1111 ii ii11111 i ~Il1l V. 9 3223 cttaatgttgtcactattgaagaaactaaggcagatgatgttgacagtgatggaaacaga 3282 V.1 :3552 gtaatgctcttgttgaggaaatgaatcatgt 3611 V.9: 3283 gtcacactagaccttcctattgatctagaagagcaaacaatgggaaagtacaattgggta 3342 V.1 :3612 acaacatctcaccaacctatgccaatcattc 3671 V. 9 :3343 acaacatctcactaacctatgccaatcattc 3402 V.1 :3672 tccaactctcaatactgaccctatcagacct 3731 V.9 :3403 tccaactctcaatactgaccctattagacct 3462 V.1 :3732 atcaacgcccaaccttggcttattttcattc 3791 V.9 :3463 atcaacgcccaaccttggcttattttcattc 3522 V.1 :3792 tcacgtaaccaactttgcggcacatagctca 3851 V.9 :3523 tcacgtaaccaactttgcggcacataactca 3582 V.1: 3852 gtacctgtgtccgtacacaccagaccg 3878 V.9 :3583 gtacctgtgtccgtacacaccagaccg 3609 Table LIV(h). Peptide sequences of protein coded by 109P1 D4 v.9 (SEQ ID NO: 282) MTVGFNSDIS SVVRVNTTNC HKCLLSGTYI FAVLLVCVVF HSGAQEKNYT IREEIPENVL IGNLLKDLNL SLIPNKSLTT TMQFKLVYKT GDVPLIRIEE DTGEIFTTGA RIDREKLCAG 120 IPRDEHCFYE VEVAILPDEI FRLVKIRFLI EDINDNAPLF PATVINISIP ENSAINSKYT 180 00 0 0 ci

C)

Co 0 0 ci 0 ci ci 00 0 0 ci

LPAAVDPOVG

KVEDGGFPQR

ENAKIHFSFS

HVIJVNVTDVN

TDHEIPFRLR

NDNAPVFTQS

GMLTVVKKLD

PENLPRHGTV

TFYVKAEDGG

AVDNDTGb4NA

DSIJFSVVIVN

GTITVVVVIF

NLLLNVVTIE

KSAS POPAFO T FEVP VS VHT

INGVQNYELI

SSTAILQVSV

NLVSNIARRL

DNVPSIDIRY

PVFSNQFLLE

FVTVSIPENN

REKEDKYLFT

GLITVTDPDY

RVSRSSSAKV

EVRYS IVGGN

LFVNESVTNA

ITAVVRCRQA

ETKADDVDS 0

IQPETPLNLK

RPTDSRT

KSQNI FGLDV

TDTNDNHPVF

FHLNATTGLI

IVNPVNDTVV

NAAYLDYEST

S FGIQLMKVS

ILAKDNGVPP

GDNSAVTLSI

TINVVDVNDN

TRDLFAIDQE

TLINELVRKS

PHLKAAQKNM

GNRVTLDLPI

HHIlIQELPLD

IETPEGDKMP

KETEIEVSIP

T IKE PtORE E LSENI PLNTK

KEYAIKIJIAA

ATDADSGPNA

LTSNVTVFVS

LDENDDFTID

KPVFIVPPYN

TGNITLMEKC

IEAPVTPNTE

QNSBWATPNP

DLEEQTMGKY

NTFVACDSIS

QLIVQKELDR

ENAPVGTSVT

TPNHKLLVLA

IALITVTDKD

DAGKPPLNQS

EINYLLGPDA

IIIJQNDNSPV

SQTGVIRPNI

YS YEL Vt PST

DVTDLGLHRV

IADVSSPTSD

ENRQMIMMKK

NWVTTPTTFK

NCSSSSS OPY

EEKDTYVMKV

QLHATDADIG

SDGGLMPARA

ADHNGRVTCF

AMLFIKVKDE

PPEFSLDRRT

FTHNEYKFYV

SFDREKQESY

NPGTVVFQVI

LVKANDLGQP

YVKILVAAVA

KKKKKKHS 2K

PDSPDLARHY

SVS DCGYPVT 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1037 Table LV(h). Amino add sequence alignment of 1 09P1 D4 v.1 (SEQ ID NO: 28) and 109P1ID4 v.9 (SEQ ID NO: 284) Score =1961 bit (5081), Expect =O.Oldenflfles 99211009 Posiflves; 995/1109 (98%) V. 1 3 LLSGTYIFAVLLACVVFHSGAENYTIREEMPENVLIGDLIKDLNLSLIPKSLTTAMQ 62 LLSGTYIFAVLL CVVFHSGAQEKNY'IREE+PENVLIG+LLKDLNLSLIPNKSLTT

MO

V.9 24 LLSGTYIFAVLLVCVVFHSGAEKNTIREEIPENVLIGNLLKDLNLSLIPNSLTTTMQ 83 V.1 63 FKLVYKTGDVPLIRIEEDTGEITTGARIDREnLCAGIPRDEHCFYEVEVMILPDEIFRL 122 PnVYKTGDVPLIRIEEDTGEIFTTGARIDREKLCAGIPRDEHC!'YEEVAILPDEIFRL V.9 84 FKLVYKTGDVPLIRIEEDTGEIFTTGARIDREKLCAGIPRDEHCFYEVEVAILPDEIflL 143 V.1 123 VKIRFLIEDINDNAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVONYELIRSQ 182 VKIRFLIEDINDNAPLFPATVINISIPENSAINSICYTLPAAVDPDVGINGVQNYhLIKSQ V.9 144 VKIRFLIEDINDNAPLFPATVINISIPENSAINSKYTLPAAVDPDVGINGVONYELIKSQ 203 V.1 183 NI FGLDVIETPEGDKPQLIVQKELDREEKDTYVMKVKCEDGGFPORSSTMILQVSVTDT 242

NIFGLDVIETPEGDKMPQLIVQKELDREEKDTYVMKVKVEDGGFPRSSTAILQVSVTDT

V.9 204 NIFGLDVIETPEGDKNPOLIVOKELDREEKDTYVMKVKVEDGGFPRSSTMILQVSrTDT 263 V.1 ±243 NDNHPVFflTEIEVSIPENAPVGTSVTLHATDADIGENKIHFSFSNLVSNIRLSR 302

NDNHPVFKETEIEVSIPENAPVGTSTLHATDADIGENAKIHFSFSNLVSNIRLH

V.9 264 NDWHPVMnTEIEVSIPENAPVGTSVTOLHATDADIGENAKIFSFSNLVSNIRRFRL 323 V.1 303 NATTGLITIKEPLDREETPNHKLLVLASDGGLMPARAMLVNTDNDNVPSIDIRYIVN 362

NATTGLITIKEPLDREETPNHKLLVLASDGGAMPARAMVLVTDDPSIDIRYIVN

V.9 324 NATTGLITIKEPLDREETPNHKLLVLASDGGLMPARAMVLVNVTDVNDNVPSIDIRYIVN 383 V.1 363 PVNDTVVLSENIPLNTKIAJITVTDKDADHNGRVTCFTDHEIPFRLRPVFSNQFLISTAA 422 PVNDTVVLSENI PLNTKIMJITVTDKDADHNGRVTCETDHEIPFRLRPVFSNQFLLE

AA

V.9 384 PVNDTVVLSENI PLNTKIALITVTDKDADHNGRVTCfDEIPFRRPVFSNQFLLENAA 443 V.1 423 YLDYESTKEYAIKLLADAGKPPLNQSAMLFIKKDENDNAPVEQSFVTVS IPENS P 482 YLDYESTKEYAIKLLAADAKPPLNQSMFIKKDENDNAPV'QSFVTVSI

PENNSPG

V.9 444 YLDYEsTKEYAIKLLAAnACKPPLNOSAMLFIKlcDEN DNAPVflS EVTVSI PENNSPG 503 V.1 483 IOLTKVSAMDADSGPNAKINYLLGPDAPPEFSLDCRTGMLTVVLDRKEDLflIIA 542 IQL KVSA DADSGPNA+INYLLGPDAPPEFSLD RTGMLTVVKKIJDREKEDKYLFl'ILA V.9 504 IQI 2 (VSATDADSGPNAEINYLLGPDAPPEFSLDRRTGMLTVVKLDREKEDYLTIIA 563 V.1 ±543 KDNGVPPLTSNVTVFVSIIDQNDNSPVFTHNEYNFYVPENLPRHGTVGLITVDPDYGDN 602 ICDNGVPPLTSNVTVFVSIIDONDNSPVFHNEY

FYVPENLPRHGTVGLITVTDPDYCON

V.9 ±564 KDNGVPPLTSNVTVFVSIIDQNDNSPVF7HNEYKFYVPENLPRHGTVGLITVTDPDYGDN 623 V.1 603 SAVTLSILDENDDFTIDSQTGVIRPNISFDREKQESYTFYVEGGRVSRSSSKVIN 662

SAVTLSILDENDDFIDSQTGVIRPNISFDREKQESYTFYVKAEDGGRVSRSSSAKVTIN

V.9 624 SAVTLSILDENDDFrIDSQTGVIRPNISFDREKQESYTFYVKAEGGRVSRSSSAKVTIN 683 V.1 663 VVDVNDNKPVFIVPPSNCSYELVLPSTNPGTWVFQVIAVDNDTGNNAEVRYSIVGGNTRD 722 VVDVNDNKPVFIVPP N SYELVLPSTNMGTVVFQVIAVDNDTGMNAEVRYSIVGGNTRD V.9 684 WVDVNDNIKPVFIVPPYNYSYELVLPSTNPGTVVFQVIAVDNDTGAEVRYSIVGGNTRD 743 303 00 OV.1 723 LFAIDQETGNITLMEKCDVTDLGLHRVLVKANDLGQPDSLFSVVIVNLFNESVTNATLI 782

OLFAIDQETGNITLMEKCDVTDLGLHRVLVKANDLGPDSLFSIVNLVESVTNATLI

V.9 744 LFAIDOETGNITLMEKCDVTDLGLRVLVKANDLGQPDSLFSVVIVNLFNESVNATLI 803 V.1 783 NELVRKSTEAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVIFITAVCROAPHL 842 C.)NELVRKS EAPVTPt4TEIADVSSPTSDYVKILVAAVATITVVIFITAVRCQAPHL CCV.9 804 NELVRKSIEAPVTPNTEIADVSSPTSDYVKILVAAVAGTITVVVIFITAVR HL 863 oV.1 843 KAAQI KQNSEWATPNPENROMI GUG KKKKHSPKNLLLNFVTIEETKADDVDSDGNR 902 KAAQKN QNSEWATPNPENRQMIMMI{KKKKKKKHSPKNLLLN

VTIEETKADDVDSDGNR

V.9 864 KAAQKNMQNSEWATPNPEWRQMIM (10 WKKKSPKNLLLNVIETKCADDVDSDN 923 OV.1 903 VTLDLPIDLEEQTMGKYNWVTTPTFKPDSPDLARHYKSASPPAFQIQPETPLNSKHI 962 cIVTLDLPIDLEEQTMGKYNWVTTITFKPDSPDLARHYKSASPQPAFQIQPETPLN

KH

OV.9 924 VTLDLPIDLEEQTMGKYI4WVTTPTTFKPDSPDLARHYKSASPQPAFQIQPETPLNLRHI 983 V. 6 QLLNFADIKSSSDYVDGPTiVVVTP11 V.1 963IQELPLDNTFVACDSI5KCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP 11 00 .9 98 IQELPLDNTFVACDSISNCSSSSSDPYSVSDCGYPVTTFEVPVSVHTRP 13

Claims (39)

1. A composition that comprises a) a peptide of eight, nine, ten, or eleven contiguous amino acids of a protein of Figure 2 b) a peptide of Tables VIII-XXI c) a peptide of Tables XXII to XLV or, d) a peptide of Tables n XLVI to XLIX. O

2. A composition of claim 1, which elicits an immune response.

3. A protein of claim 2 that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% homologous or identical to an Sentire amino acid sequence shown in Figure 2. C 4. A protein of claim 2, which is bound by an antibody that specifically binds to a protein of Figure 2. 00 A composition of claim 2 wherein the composition comprises a cytotoxic T cell (CTL) polypeptide epitope S 10 or an analog thereof, from the amino acid sequence of a protein of Figure 2.

6. A composition of claim 5 further limited by a proviso that the epitope is not an entire amino acid sequence of Figure 2.

7. A composition of claim 2 further limited by a proviso that the polypeptide is not an entire amino acid sequence of a protein of Figure 2.

8. A composition of claim 2 that comprises an antibody polypeptide epitope from an amino acid sequence of Figure 2.

9. A composition of claim 8 further limited by a proviso that the epitope is not an entire amino acid sequence of Figure 2. A composition of claim 8 wherein the antibody epitope comprises a peptide region of at least 5 amino acids of Figure 2 in any whole number increment up to the end of said peptide, wherein the epitope comprises an amino acid position selected from a) an amino acid position having a value greater than 0. 5 in the Hydrophilicity profile of Figure 5, b) an amino acid position having a value less than 0. 5 in the Hydropathicity profile of Figure 6; c) an amino acid position having a value greater than 0. 5 in the Percent Accessible Residues profile of Figure 7 d) an amino acid position having a value greater than 0. 5 in the Average Flexibility profile of Figure 8; e) an amino acid position having a value greater than 0. 5 in the Beta-tum profile of Figure 9 f) a combination of at least two of a) through e) g) a combination of at least three of a) through e) h) a combination of at least four of a) through e) or i) a combination of five of a) through e).

11. A polynucleotide that encodes a protein of claim 1.

12. A polynucleotide of claim 11 that comprises a nucleic acid molecule set forth in Figure 2.

13. A polynucleotide of claim 12 further limited by a proviso that the encoded protein is not an entire amino acid sequence of Figure 2. 00 305 O

14. A composition comprising a polynucleotide that is fully complementary to a polynucleotide of claim 11. L) 15. An 109P1 04 siRNA composition that comprises siRNA (double stranded RNA) that corresponds to the nucleic acid ORF sequence of the 109P1D4 protein or a subsequence thereof; wherein the subsequence is 19, 21, 22, 23, 24, or 25 contiguous RNA nucleotides in length and contains sequences that are complementary and non- complementary to at least a portion of the mRNA coding sequence. C 16. A polynucleotide of claim 13 that further comprises an additional nucleotide sequence that encodes an additional peptide of claim 1.

17. A method of generating a mammalian immune response directed to a protein of Figure 2, the method comprising exposing cells of the mammal's immune system to a portion of a) a 109P1 D4-related protein and/or b) C 10 a nucleotide sequence that encodes said protein, whereby an immune response is generated to said protein.

18. A method of generating an immune response of claim 17, said method comprising providing a 109P1 D4- related protein that comprises at least one T cell or at least one B cell epitope and, contacting the epitope with a mammalian immune system T cell or B cell respectively, whereby the T cell or B cell is activated.

19. A method of claim 18 wherein the immune system cell is a B cell, whereby the activated B cell generates antibodies that specifically bind to the 109P1 D4-related protein. A method of claim 18 wherein the immune system cell is a T cell that is a cytotoxic T cell (CTL), whereby the activated CTL kills an autologous cell that expresses the 109P1D4-related protein.

21. A method of claim 18 wherein the immune system cell is a T cell that is a helper T cell (HTL), whereby the activated HTL secretes cytokines that facilitate the cytotoxic activity of a cytotoxic T cell (CTL) or the antibody- producing activity of a B cell. corresponds to the nucleic acid ORF sequence of the 109P1D4 protein or a subsequence thereof; wherein the subsequence is 19, 20, 21, 22, 23, 24, or 25 contiguous RNA nucleotides in length and contains sequences that are complementary and non-complementary to at least a portion of the mRNA coding sequence. A composition of claim 28, further comprising a physiologically acceptable carrier.

31. A pharmaceutical composition that comprises the composition of claim 28 in a human unit dose form.

32. A composition of claim 28 wherein the substance comprises an antibody or fragment thereof that specifically binds to a protein of Figure 2.

33. An antibody or fragment thereof of claim 32, which is monoclonal.

34. An antibody of claim 32, which is a human antibody, a humanized antibody or a chimeric antibody.

35. A non-human transgenic animal that produces an antibody of claim 32. 00 306

36. A hybridoma that produces an antibody of claim 33. )J 37. A composition of claim 28 wherein the substance reduces or inhibits the viability, growth or reproduction status of a cell that expresses a protein of Figure 2. O

38. A composition of claim 28 wherein the substance increases or enhances the viability, growth or reproduction status of a cell that expresses a protein of Figure 2.

39. A composition of claim 28 wherein the substance is selected from the group comprising a) an antibody or fragment thereof, either of which immunospecifically binds to a protein of Figure 2 b) a polynucleotide that encodes 00 an antibody or fragment thereof, either of which immunospecifically binds to a protein of Figure 2; c) a ribozyme that 00 cleaves a polynucleotide having a 109P1 D4 coding sequence, or a nucleic acid molecule that encodes the 10 ribozyme; and, a physiologicaliy acceptable carrier; and d) human T cells, wherein said T cells specifically recognize a 109P1 D4 peptide subsequence in the context of a particular HLA molecule e) a protein of Figure 2, or a fragment of a protein of Figure 2 a nucleotide encoding a protein of Figure 2, or a nucleotide encoding a fragment of a protein of Figure 2 g) a peptide of eight, nine, ten, or eleven contiguous amino acids of a protein of Figure 2; h) a peptide of Tables VIII-XXI i) a peptide of Tables XXII to XLV; j) a peptide of Tables XLVI to XLIX; k) an antibody polypeptide epitope from an amino acid sequence of Figure 2 I) a polynucleotide that encodes an antibody polypeptide epitope from an amino acid sequence of Figure 2 or m) an 109P1 D4 siRNA composition that comprises siRNA (double stranded RNA) that corresponds to the nucleic acid ORF sequence of the 109P1 D4 protein or a subsequence thereof wherein the subsequence is 19, 20, 21, 22, 23, 24, or 25 contiguous RNA nucleotides in length and contains sequences that are complementary and non-complementary to at least a portion of the mRNA coding sequence. A method of inhibiting viability, growth or reproduction status of cancer cells that express a protein of Figure 2, the method comprising administering to the cells the composition of claim 28, thereby inhibiting the viability, growth or reproduction status of said cells.

41. The method of claim 40, wherein the composition comprises an antibody or fragment thereof, either of which specifically bind to a 109P1 D4-related protein.

42. The method of claim 40, wherein the composition comprises a 109P1 D4-related protein or, (ii) a polynucleotide comprising a coding sequence for a 109P1 D4-related protein or comprising a polynucleotide complementary to a coding sequence for a 109P1 D4-related protein.

43. The method of claim 40, wherein the composition comprises a ribozyme that cleaves a polynucleotide that encodes a protein of Figure 2.

44. The method of claim 40, wherein the composition comprises human T cells to said cancer cells, wherein said T ceils specifically recognize a peptide subsequence of a protein of Figure 2 while the subsequence is in the context of the particular HLA molecule. 00 307

45. The method of claim 40, wherein the composition comprises a vector that delivers a nucleotide that e( encodes a single chain monoclonal antibody, whereby the encoded single chain antibody is expressed intracellularly )within cancer cells that express a protein of Figure 2.

46. A method of delivering an agent to a cell that expresses a protein of Figure 2, said method comprising providing the agent conjugated to an antibody or fragment thereof of claim 32 and, exposing the cell to the antibody-agent or fragment-agent conjugate.

47. A method of inhibiting viability, growth or reproduction status of cancer cells that express a protein of Figure 2, the method comprising: administering to the cells the composition of claim 28, thereby inhibiting the 00 viability, growth or reproduction status of said cells. 10 48. A method of targeting information for preventing or treating a cancer of a tissue listed in Table I to a subject in need thereof, which comprises detecting the presence or absence of the expression of a polynucleotide associated with a cancer of a tissue listed in Table I in a sample from a subject, wherein the expression of the polynucleotide is selected from the group consisting of: a nucleotide sequence in Figure 2 a nucleotide sequence which encodes a polypeptide encoded by a nucleotide sequence in Figure 2 a nucleotide sequence which encodes a polypeptide that is 90% or more identical to the amino acid sequence encoded by a nucleotide sequence in Figure 2 directing information for preventing or treating the cancer of a tissue listed in Table I to a subject in need thereof based upon the presence or absence of the expression of the polynucleotide in the sample.

49. The method of claim 48, wherein the information comprises a description of detection procedure or treatment for a cancer of a tissue listed in Table 1.

50. A method for identifying a candidate molecule that modulates cell proliferation, which comprises (a) introducing a test molecule to a system which comprises a nucleic acid comprising a nucleotide sequence selected from the group consisting of: the nucleotide sequence of SEQ ID NO 1 (ii) a nucleotide sequence which encodes a polypeptide consisting of the amino acid sequence set forth in Figure 3 (iii) a nucleotide sequence which encodes a polypeptide that is 90% or more identical to the amino acid sequence set forth in Figure 3 and (iv) a fragment of a nucleotide sequence of or (iii) or introducing a test molecule to a system which comprises a protein encoded by a nucleotide sequence of (iii), or (iv) and determining the presence or absence of an interaction between the test molecule and the nucleotide sequence or protein, whereby the presence of an interaction between the test molecule and the nucleotide sequence or protein identifies the test molecule as a candidate molecule that modulates cell proliferation.

51. The method of claim 50, wherein the system is an animal.

52. The method of claim 50, wherein the system is a cell.

53. The method of claim 50, wherein the test molecule comprises an antibody or antibody fragment that specifically binds the protein encoded by the nucleotide sequence of (iii), or (iv). 00 308

54. A method for treating a cancer of a tissue listed in Table I in a subject, which comprises administering a Scandidate molecule identified by the method of claim 50 to a subject in need thereof, whereby the candidate Smolecule treats a cancer of a tissue listed in Table I in the subject. A method for identifying a candidate therapeutic for treating a cancer of a tissue listed in Table 1, which comprises: introducing a test molecule to a system which comprises a nucleic acid comprising a nucleotide sequence selected from the group consisting of: the nucleotide sequence of SEQ ID NO 1 (ii) a nucleotide sequence which encodes a polypeptide consisting of the amino acid sequence set forth in Figure 3; (iii) a nucleotide sequence which encodes a polypeptide that is 90% or more identical to the amino acid sequence set forth in Figure C~ 3 and (iv) a fragment of a nucleotide sequence of or (iii) or introducing a test molecule to a system which 00 0 10 comprises a protein encoded by a nucleotide sequence of (iii), or (iv} and determining the presence or 0. absence of an interaction between the test molecule and the nucleotide sequence or protein, whereby the presence of an interaction between the test molecule and the nucleotide sequence or protein identifies the test molecule as a candidate therapeutic for treating a cancer of a tissue listed in Table 1.

56. The method of claim 55, wherein the system is an animal.

57. The method of claim 55, wherein the system is a cell.

58. The method of claim 55, wherein the test molecule comprises an antibody or antibody fragment that specifically binds the protein encoded by the nucleotide sequence of (iii), or (iv). Dated this FIFTH day of SEPTEMBER, 2008 AGENSYS, INC. by FB Rice Co Patent Attorneys