IL164326A - Cancer-associated products and antibodies binding the same - Google Patents

Abstract

A novel gene (designated 213P1F11) and its encoded protein, and variants thereof, are described wherein 213P1F11 exhibits tissue specific expression in normal adult tissue, and is aberrantly expressed in the cancers listed in Table I. Consequently, 213P1F11 provides a diagnostic, prognostic, prophylactic and/or therapeutic target for cancer. The 213P1F11 gene or fragment thereof, or its encoded protein, or variants thereof, or a fragment thereof, can be sued to elicit a humoral or cellular immune response; antibodies or T cells reactive with 213P1F11 can be used in active or passive immunization.

Description

CANCER-ASSOCIATED PRODUCT AND ANTIBODIES BINDING THE SAME 164,326/2 FIELD OF THE INVENTION The invention described herein relates to a cancer associated protein, antibodies recognizing the same and uses thereof.

It is to be noted that only subject matter embraced in the scope of the claims appended hereto, whether in the manner defined in the claims or in a manner similar thereto and involving the main features as defined in the claims, is intended to be included in the scope of the present invention, while subject matter described and exemplified to provide background and better understanding of the invention, is not intended for inclusions as part of the present invention.

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 nave 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 Northern 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 lias been a very useful tool, however its specificity mid general utility is widely regarded as lacking in several important respects.

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 LAPC (Los Angeles Prostate Cancer) xenografts are prostate cancer xenografts that have survived passage in severe combined immune deficient (SCID) mice and have exliibited the capacity to mimic the transition from androgen dependence to androgen independence (Klein er a/., 1997, Nat. Med. 3 :402). More recently identified prostate cancer markers include PCTA-1 (Su er a/. , 1996, Proc. Natl. Acad. Sci. USA 93 : 7252), prostate-specific membrane (PSM) antigen (Pinto et ai, Clin Cancer Res 1996 Sep 2 (9): 1445-51), STEAP (Hubert, et at., Proc Natl Acad Sci U S A. 1999 Dec 7; 96(25): 14523-8) and prostate stem cell antigen (PSCA) (Reiter r al , 1998, Proc. Natl. 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 furt er improve diagnosis and Uierapy.

Renal cell carcinoma (RCC) accounts for approximately 3 percent of adult malignancies. Once adenomas reach a diameter of 2 to.3 cm, malignant potential exists. In the adult, the two principal malignant renal tumors are renal cell adenocarcinoma and transitional cell carcinoma of the renal pelvis or ureter. The incidence of renal cell adenocarcinoma is estimated at more titan 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 Umted States.

Surgery has been the primary therapy for renal cell adenocarcinoma 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 aggressi vely 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 older population. In 1998, there was mi estimated 54,500 cases, including 39,500 in men mid 15,000 in women. The age-adjusted incidence in the United States is 32 per 100,000 for men and 8 per 100,000 in women. The historic male/female ratio of 3 : 1 may be decreasing related to smoking patterns in women. There were mi estimated 11,000 deaths from bladder cancer in 1998 (7,800 in men mid 3,90.0 in women). Bladder cmicer incidence and mortality strongly increase with age and will be an increasing problem as the population becomes more elderly.

Most bladder cancers recur in tire bladder. Bladder cancer is managed with a combination of transurethral resection of the bladder (TUR) mid intravesical chemotherapy or immunotherapy. The multifocal mid recurrent nature of bladder cancer points out Hie limitations of TUR. 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 patients.

An estimated 130,200 cases of colorectal cancer occurred in 2000 in tire United States, including 93,800 cases of colon cancer and 36,400 of rectal cancer. Colorectal cancers are tlie third most common cancers in men and women. Incidence rates declined significantly during 1992-1996 (-2.1% per year).

Research suggests that these declines have been due to increased screening and polyp removal, preventing progression of polyps to invasive cancers. There were an estimated 56,300 deatlis (47,700 from colon cancer, 8,600 from rectal cancer) in 2000, accounting for about 11% of all U.S. cancer deatlis.

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

There were an estimated 164,100 new cases of lung and bronchial cancer in 2000, accounting for 14% of all U.S. cancer diagnoses. The incidence rate of lung and bronchial cancer is declining significantly in men, from a high of 86.5 per 100,000 in 1984 to 70.0 in 1996. In tl e 1990s, tlie rate of increase among women began to slow. In 1996, tlie incidence rate in women was 42.3 per 100,000.

Lung and bronchial cancer caused an estimated 156,900 deatlis in 2000, accounting for 28% of all cancer deatlis. During 1992-1996, mortality from lung cancer declined significantly among men (-1.7% per year) while rates for women were still significantly increasing (0.9% per year). Since 1987, more women have died each year of lung cancer Uian breast cancer, which, for oyer 40 years, was tlie major cause of cancer death in women. Decreasing lung cancer incidence and mortality rates most likely resulted from decreased smoking rates over tl e previous 30 years; however, decreasing smoking patterns among women lag behind those of men. Of concern, although d e declines in adult tobacco use have slowed, tobacco use in youth is increasing again.

Treatment options for lung and bronchial cancer are detennined by the type and stage of tlie cancer and include surgery, radiation tlierapy, and chemotherapy. For many localized cancers, surgery is usually tlie treatment of choice. Because the disease has usually spread by the time it is discovered, radiation tlierapy and chemotherapy are often needed in combination with surgery. Chemotherapy alone or combined with radiation is tlie 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 breasl cancer were expected to occur among women in the United States during 2000. Additionally, about 1,400 new cases of breasl cancer were expected to be diagnosed in men in 2000. After increasing about 4% per year in tlie 1 80s, breast cancer incidence rales in women have leveled off in tlie 1990s to about 1 10.6 cases per 100,000.

In tlie 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 deatlis in women. According to tlie most recent dala, mortality rates declined significantly during 1992-1996 with tlie largest decreases in younger women, both white and black. These decreases were probably the result of earlier detection and improved treatment. 1 4,326/2 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.

There were an estimated 23,100 new cases of ovarian cancer in the United States in 2000. It accounts for 4% of all cancers among women and ranks second among gynecologic cancers. During 1992-1996, ovarian cancer incidence rates were significantly declining. Consequent to ovarian cancer, there were an estimated 14,000 deaths in 2000. Ovarian cancer causes more deaths than any other cancer of the female reproductive system.

Surgery, radiation therapy, and chemotherapy are treatment options for ovarian cancer. Surgery usually includes the removal of one or both ovaries, fallopian tubes (salpingo-oophorectomy), and the uterus (hysterectomy). In some very early tumors, only the involved ovary will be removed, especially in young women who wish to have children. In advanced disease, an attempt is made to remove all intraabdominal disease to enhance the effect of chemotherapy. There continues to be an important need for effective treatment options for ovarian cancer.

There were an estimated 28,300 new cases of pancreatic cancer in the United States in 2000. Over the past 20 years, rates of pancreatic cancer have declined in men. Rates among women have remained approximately constant but may be beginning to decline. Pancreatic cancer caused an estimated 28,200 deaths in 2000 in the United States. Over the past 20 years, there has been a slight but significant decrease in mortality rates among men (about-0.9% per year) while rates have increased 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 significant need for additional therapeutic and diagnostic options for pancreatic cancer.

Israeli Application Number 166,564 is a national stage entry of International Patent Application Number WO 2003258269 and is directed to the 273P4B7 gene and its encoded proteins. Israeli Application Number 166,532 is directed to the 202P5A5 gene and its encoded proteins. Israeli Patent Application Number 150,873 is a national stage entry of International Patent Application Number WO 2001237973, which corresponds to U.S. application, Serial No. 09/771,312 filed on the same day and claiming priority to the same U.S. provisional application (Serial No. 60/178,560), and is directed to the 84P2A9 gene and its encoded proteins. The instant application is directed to the 213P1F11 gene, which is separate and distinct from the aforementioned genes. 164,326/2 SUMMARY OF THE INVENTION In some embodiments, this invention provides an isolated polynucleotide that encodes a protein comprising the polypeptide sequence shown in SEQ ID NO: 18, 19, 20, 21 , 22, or 23.

In some embodiments, this invention provides a polynucleotide selected from the group consisting of: (a) a polynucleotide comprising the sequence of SEQ ID NO:2, from nucleotide residue numbers 404 through 1 132; (b) a polynucleotide comprising the sequence of SEQ ID NO:4, from nucleotide residue numbers 404 through 1096; (c) a polynucleotide comprising the sequence of SEQ ID NO:6, from nucleotide residue numbers 404 through 844; (d) a polynucleotide comprising the sequence of SEQ ID NO:8, from nucleotide residue numbers 1 through 966; (e) a polynucleotide comprising the sequence of SEQ ID NO: 10, from nucleotide residue numbers 404 through 1 132; (f) a polynucleotide comprising the sequence of SEQ ID NO:2, from nucleotide residue numbers 404 through 1 132, wherein the nucleotide residue at 2027 is T; (g) a polynucleotide comprising the sequence of SEQ ID NO:2, from nucleotide residue numbers 404 through 1 132, wherein the nucleotide residue at 2037 is C; (h) a polynucleotide comprising the sequence of SEQ ID NO:2, from nucleotide residue numbers 404 through 1 132, wherein the nucleotide residue at 2268 is G; and (i) a polynucleotide comprising the sequence of SEQ ID NO:2, from nucleotide residue numbers 404 through 1 132, wherein the nucleotide residue at 3196 is T.

In some embodiments, this invention provides a recombinant expression vector comprising a polynucleotide as herein described and in some embodiments, the vector is a viral vector. In some embodiments, the invention provides a cell containing the vector.

In some embodiments, the invention provides a process for producing a protein comprising culturing a host cell containing a vector as herein described under conditions sufficient for the production of the protein, wherein the amino acid sequence of the protein is selected from the group consisting of SEQ ID NO: 18, 19, 20, 21 , 22, and 23.

In some embodiments, the invention provides an antibody or fragment thereof that immunospecifically binds to an epitope on a protein comprising the amino acid sequence of SEQ ID NO: 18, 19, 20, 21, 22, or 23. In some embodiments, the antibody is a human antibody. In some embodiments, the 164,326/2 antibody is labeled with a cytotoxic agent. In some embodiments, the cytotoxic agent is selected from the group consisting of radioactive isotopes, chemotherapeutic agents and toxins. In some embodiments, the cytotoxic agent is a radioactive isotope selected from the group consisting of 2l lAt, l3 lI, l251, 0Y, l86Re, l88Re, Sm, Bi, P and radioactive isotopes of Lu. In some embodiments, the cytotoxic agent is a chemotherapeutic agent selected from the group consisting of taxol, actinomycin, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, gelonin, and calicheamicin. In some embodiments, the cytotoxic agent is a toxin selected from the group consisting of diphtheria toxin, enomycin, phenomycin, Pseudomonas exotoxin (PE) A, PE40, abrin, abrin A chain, mitogellin, modeccin A chain, and alpha-sarcin.

In some embodiments, the invention provides an in vitro method for detecting the presence of a protein comprising the amino acid sequence of SEQ ID NO: 18, 19, 20, 21 , 22, or 23 or a polynucleotide comprising the polynucleotide sequence of SEQ ID NO: 4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO: 10, SEQ ID NO: 12, or SEQ ID NO:2, wherein 2027 is C or T, 2037 is T or C, 2268 is A or G, and 3196 is A or T, in a test sample comprising: contacting the sample with an antibody or polynucleotide, respectively, that specifically binds to the protein or polynucleotide, respectively; and detecting binding of protein or polynucleotide, respectively, in the sample thereto.

In some embodiments, the invention provides an in vitro method of inhibiting growth of a cell expressing a protein comprising the amino acid sequence of SEQ ID NO: 18, 19, 20, 21 , 22, or 23, comprising providing an effective amount of an antibody according to any one of claims 1 1 to 22 to the cell, whereby the growth of the cell is inhibited.

In some embodiments, the invention provides an in vitro method of delivering a cytotoxic agent to a cell expressing a protein comprising the amino acid sequence of SEQ ID NO: 18, 19, 20, 21 , 22, or 23, comprising providing an effective amount of an antibody as herein described.

In some embodiments, the invention provides for the use of a protein comprising the amino acid sequence of SEQ ID NO: 18, 19, 20, 21 , 22, or 23, for the preparation of a medicament to induce an immune response from a subject, whereby a T cell or B cell is activated.

In some embodiments, this invention provides for the use of an effective amount of antibody or antigen binding fragment thereof for the preparation of a medicament which delivers a cytotoxic agent to a cell expressing a protein comprising the amino acid sequence of SEQ ID NO: 18, 19, 20, 21 , 22, or 23, wherein the antibody or antigen binding fragment thereof comprises the antibody according to any one of claims 17 to 21. 164,326/1 BRIEF DESCRIPTION OF THE FIGURES Figure 1. The 213P1F11 SSH sequence of 166 nucleotides.

Figure 2. The cDNA and amino acid sequence of 213P1F11 variant 1 clone CASP14-BrCl (also called "213P1F11 v.1" or "213P1F11 variant 1" or "213P1F11") is shown in Figure 2A. The start methionine is underlined. The open reading frame extends from nucleic acid 404-1132 including the stop codon. The cDNA and amino acid sequence of 213P1F11 variant 2 (also called "213P1F11 v.2") is shown in Figure 2B. The codon for the start metluonine is underlined. The open reading frame extends from nucleic acid 409-1096 including the stop codon. The cDNA and amino acid sequence of 213P1F11 variant 3 (also called "213P1F11 v.3") is shown in Figure 2C. The codon for the start met onine is underlined. The open reading frame extends from nucleic acid 404-844 including the stop codon. The cDNA and amino acid sequence of 213P1F11 variant 4 (also called "213P1F11 v.4") is shown in Figure 2D. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 1-966 including the stop codon. The cDNA (SEQ ID. NO. and amino acid sequence of 213P1F11 variant 5 (also called "213P1F11 v.5") is shown in Figure 2E. The codon for the start metliionine is underlined. The open reading frame extends from nucleic acid 404-1132 including the stop codon. The cDNA and amino acid sequence, of 213P1F11 variant 6 (also called "213P1F11 variant v.6") is shown in Figure 2F. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 404-1132 including the stop codon. The cD A and amino acid sequence of 213P1F11 variant 7 (also called "213P1F11 v.7") is shown in Figure 2G. The codon for the start methionine is underlined. The open reading frame extends from nucleic acid 404-1132 including the stop codon. The cDNA and airiino acid sequence of 213P1F11 variant 8 (also called "213P1F11 v.8") is shown in Figure 2H. The codon for the start meunonine is underlined. The open reading frame extends from nucleic acid 404-1132 mcluding the stop codon. As used herein, a reference to 213P1F11 includes all variants thereof, including those shown in Figure 10. 6a Figure 3. Amino acid sequence of 213P1F11 v. l is shown in Figure 3 A; it lias 242 amino acids. The amino acid sequence of 213P1F11 v.2 is shown in Figure 3B; it has 230 amino acids. The amino acid sequence of 213P1F1 1 v.3 is shown in Figure 3C; it has 146 amino acids. The amino acid sequence of 213P1F11 v.4 is shown in Figure 3D; it has 321 amino acids. The amino acid sequence of 213P 1F11 v.5 is shown in Figure 3E; it has 242 amino acids. The amino acid sequence of 213P1F11 v.6 is shown in Figure 3F; it has 242 amino acids. As used herein, a reference to 213P1F11 includes all variants thereof, including those shown in Figure 11.

Figure 4. The nucleic acid sequence alignment of 213P1F11 v. l with human Caspase-14 (gi 6912286) precursor mRNA is shown in Figure 4A. The amino acid sequence alignment of 213P1 F11 v. l with human Caspase-14 (gi 6912286) mRNA is shown in Figure 4B. The amino acid sequence alignment of 213P 1 F1 1 v. l with mouse Caspase-14 (gi 6753280) mRNA is shown in Figure 4C. The amino acid sequence alignment of 213P1F11 v.2 with human Caspase-14 (gi 6912286) mRNA is shown in Figure 4D. The amino acid sequence alignment of 2 13P1F1 1 v.3 with human Caspase-14 (gi 6912286) mRNA is shown in Figure 4E. The amino acid sequence alignment of 213P1F11 v.2 widi mouse caspase 14 (gi 6753280) mRNA is shown in Figure 4F. The amino acid sequence alignment of 213P1F11 v.4 with human Caspase-14 (gi 6912286) mRNA is shown in Figure 4G.

Figure 5. Hydrophilicity amino acid profile of A) 213P1F11 variant 1 , B) 213P1 F11 variant 2, C) 213P1F11 variant 3 and D) 213P1F11 variant 4, determined by computer algorithm sequence analysis using the method of Hopp and Woods (Hopp T.P., Woods K.R., 1981. Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828) accessed, on tlie Protscale website located at tlie World Wide Web (.expasy.cli cgi-bin/protscale.pl) tlirough tl e ExPasy molecular biology server.

Figure 6. Hydropathicity amino acid profile of A) 213P1F11 variant 1 , B) 213P1 F11 variant 2, C) 213P1F 11 variant 3 mid D) 213P1F1 1 variant 4, determined by computer algorithm sequence analysis using Uie method of yte and Doolittle (Kyte J., Doolittle R.F., 1982. J. Mol. Biol. 157: 105-132) accessed on the ProtScale website (www.expasy.ch/cgi-bin/protscale.pl) tlirough tlie ExPasy molecular biology server.

Figure 7. Percent accessible residues amino acid profile of A) 213P1F11 variant 1, B) 213P1F11 variant 2, C) 213P1F11 variant 3 and D) 213P1F11 variant 4, deteniiined by computer algoritlim sequence analysis using tlie method of Janin (Janin J., 1979 Nature 277:491-492) accessed on tlie ProtScale website located at die World Wide Web (.expasy.cli/cgi-bin/protscale.pl) tlirough tlie ExPasy molecular biology server.

Figure 8. Average flexibility amino acid profile of A) 213P1F11 variant 1, B) 213P1F11 variant 2, C) 213P1F11 variant 3 and D) 213P1F11 variant 4, determined by computer algorithm sequence analysis using tlie method of Bhaskaran and Ponnuswamy (Bhaskaran R., and Ponnuswamy P.K.., 1988. Int. J. Pept. Protein Res. 32:242-255) accessed on (l e ProtScale website located at the World Wide Web (.expasy.cli/cgi-bin/protscale.pl) tlirough tlie ExPasy molecular biology server.

Figure 9. Beta-turn amino acid profile of A) 213P1F1 1 variant 1, B) 213P1F1 1 variant 2, C) 213P 1F1 1 variant 3 and D) 213P1F11 variant 4, determined by computer algorithm sequence analysis using tlie mediod of Deleage and Roux (Deleage, G., Roux B. 1987 Protein Engineering 1 :289-294) accessed on tlie ProtScale website located at the World Wide Web (.expasy.ch/cgi-bin/protscale.pl) dirough die ExPasy molecular biology server. 7 Figure 10. Schematic display of nucleotide variants of 213P1F1 J . Variants 213P1F11 v.2 and v.3 are splice variants. Variant 213P1F1 1 v.4 is an alternative transcript. Others are Single Nucleotide Polymorphism (also called "SNP") variants, which could also occur in any of the transcript variants that contains the base pairs. Numbers in "( )" underneath the box correspond to those of 213P1F11 v.1. The black boxes show the same sequence as 213P1F 11 v. 1. SNPs are indicated above the box.

Figure 11. Schematic display of protein variants of 213P1F11. Nucleotide variants 213P1F11 v. l though v.6 in Figure 10 code for protein variants 213P1FU v. l through 213P1F11 v.6, respectively. Variants 213P1F11 v.7 through v.10 code the same protein as variant 213P1F11 v.1. Protein variants 213P1F11 v.5 and v.6 are variants with single amino acid variations, which may exist in transcript variants 213P1F11 v.2 through 4. The black boxes show the same sequence as 213P1 F11 v. l. The numbers in "()" underneath the box correspond to those of 213P1F11 v. 1. Single amino acid differences are indicated above the box.

Figure 12. Secondary structure prediction for 213P1F1 1 variants 1 through 4. The secondary structures of 213P1F11 variant 1 (SEQ ID NO: 3) (A), variant 2 (SEQ ID NO: 5) (B), variant 3 (SEQ ID NO:7) (C), and variant 4 (SEQ ID NO: 9) (D) were predicted using the HNN - Hierarchical Neural Network method (Guenneur, 1997, . http://pbil.ibcp. fr/cgi-binynpsa_automat.pl?page=npsa_nn. html), accessed from the ExPasy molecular biology server located at the World Wide Web (.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 in a given secondary structure is also listed for each variant.

Figure 13. Exon compositions of transcript variants of 213P1F11. Variant 213P1F11 v.2 and v.3 are splice variants. Variant 213P1F11 v.4 is an alternative transcript. Compared with 213P1F1 1 v. l, 213P1 F1 1 v.2 has a longer (+74 bp at 5 ' end) exon 6 and variant 213P1F11 v.3 has a longer (+68 bp at 5' end) exon 5. Variant 213P1F1 1 v.4 has three different exons. Relative locations of exons from all variants on the clvromosome are shown at the bottom. Numbers in "( )" underneath the box correspond to those of 213P1 F1 1 v. l. Black boxes show the same sequence as 213P1F11 v. l . Intron lengths are not proportional.

Figure 14. Expression of 213P1F11 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC xenograft pool (LAPC-4AD, LAPC-4A1, LAPC-9AD and LAPC-9AI), bladder cancer pool, breast cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantilative PCR, using primers to 213P1 F11, was performed at 26 and 30 cycles of amplification. Results show strong expression of 213P1F11 in bladder cancer pool, breast cancer pool, xenograft pool, and cancer metastasis pool.

Figure 15. Expression of 213P1F11 v. l compared to 213P1F11 v.2 in patient cancer samples by RT-PCR. To determine tire relative expression of 213P1F11 v. l compared to 213P1 F1 1 v.2 in human cancers, primers were designed flanking the insertion in 213P1F 1 1 v.2. Using these primers, amplification of 213P1F11 v. l will generate a PCR fragment of 165 bp, whereas 213P1F11 v.2 will generate a PCR fragment of 249 bp as depicted in the figure. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), bladder cancer pool, breast cancer pool, LAPC xenograft pool (LAPC-4AD, LAPC-4AI, LAPC-9AD and LAPC-9AI), and 213P1FI I v.1 plasmid control. Normalization was performed by PCR using primers to actin and GAPDH. Seini-quantitative PCR, using primers depicted above, was performed at 35 cycles of amplification. Results show strong expression of 213P1 Fl 1 v.1 in bladder cancer pool, breast cancer pool, 8 LAPC xenograft pool, and the plasmid positive control. A lower expression of the 249 bp 213P1F11 v.2 product was detected in breast cancer pool, LAPC xenograft pool, and to lower extent in bladder cancer pool.

Figure 16. Expression of 213P1F11 in normal tissues. Three multiple tissue nortliern blots (A and B, Clontech; C, OriGene) widi 2 ug of mRNA/lane were probed with the 213P1F11 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show strong expression of 213P1F11 only in skin tissue. A weak transcript is detected in normal thymus but not in the other tissues tested.

Figure 17. Expression of 213P 1F11 in bladder cancer patient tissues. RNA was extracted from normal bladder (N), bladder cancer cell lines (UM-UC-3 and SCaBER), bladder cancer patient tumors (T) and normal tissue adjacent to bladder cancer (ΝΑτ). Nortliern blots with 10 ug of total RNA were probed with the 213P1 Fl 1 SSH fragment. Size standards in kilobases are indicated on the side. Results show strong expression of 213P 1F1 1 in the bladder tumor tissues but not in normal bladder nor in the bladder cancer cell lines.

Figure 18. Expression of 213P1F11 in prostate cancer xenografts. RNA was extracted from normal prostate, LAPC-4AD, LAPC-4AI, LAPC-9AD and LAPC-9AI prostate cancer xenografts. Noitliern blot with 10 ug of total RNA/lane was probed with 213P1F11 SSH sequence. Size standards in kilobases (kb) are indicated on the side. The results show expression of 213P1F11 in die LAPC-9A1 xenograft, but not in the other xenografts nor in normal prostate.

Figure 19. Expression of 213P1 Fl 1 in breast cancer patient tissues. RNA was extracted from s normal breast (N), breast cancer cell lines (DU4475, MCF7 and CAMA-1), breast cancer patient tumors (T) and breast cancer metastasis to lymph node (Met). Nortliern blots with 10 ug of total RNA were probed with die 213P1F1 1 SSH fragment. Size standards in kilobases are indicated on die side. Results show strong expression of 213P1 F1 1 in die breast tumor tissues as well as in die cancer metastasis specimen. Weak expression was also detected in die CAMA-1 cell line, but not in die odier 2 breast cancer cell lines tested.

DETAILED DESCRIPTION OF THE INVENTION Outline of Sections I. ) Definitions II. ) 213P1F11 Polynucleotides II.A.) Uses of 213P1F11 Polynucleotides II. A.1.) Monitoring of Genetic Abnormalities II.A.2.) Antisense Embodiments II.A.3.) Primers and Primer Pairs 1I.A.4.) Isolation of 213PlFll-Encoding Nucleic Acid Molecules DLA.5.) Recombinant Nucleic Acid Molecules and Host-Vector Systems HL) 213PlFll-related Proteins HI.A.) Motif-bearing Protein Embodiments HI.B.) Expression of 213PlFll-related Proteins HI.C.) Modifications of 213PlFll-related Proteins HI.D.) Uses of 213PlFll-related Proteins IV.) 213P1F11 Antibodies 9 V. ) 213P1F11 Cellular Immune Responses VI. ) 213P1F11 Transgenic Animals . ^ Vn.) Methods for the Detection of 213P1F11 VIII. ) Methods for Monitoring the Status of 213P1F11-related Genes and Their Products IX. ) Identification of Molecules That Interact With 213P1F11 X. ) Therapeutic Methods and Compositions X.A.) Anti-Cancer Vaccines X.B.) 213P1F11 as a Target for Antibody-Based Therapy X.C.) 213P1 Fll as a Target for Cellular Immune Responses X.C. I. Minigene Vaccines X.C.2. Combinations of CTL Peptides with Helper Peptides X.C.3. Combinations of CTL Peptides with T Cell Priming Agents X.C.4. Vaccine Compositions Comprising DC Pulsed with CTL and/or HTL Peptides X.D.) Adoptive Immunotherapy X. E.) Administration of Vaccines for Therapeutic or Prophylactic Purposes XI. ) Diagnostic and Prognostic Embodiments of 213P1F11.

XII. ) Inhibition of 213P1F11 Protein Function XII. A.) Inhibition of 213P1F11 With Intracellular Antibodies ΧΠ.Β.) Inhibition of 213P1F11 with Recombinant Proteins XII. C.) Inhibition of 213P1F11 Transcription or Translation XII.D.) General Considerations for Therapeutic Strategies XIH) KITS L) Definitions: Unless otherwise defined, all terms of art, notations and other scientific terms or terminology used herein are intended to have the meanings commonly understood by those of skill in the art to which this invention pertains. In some cases, tenns 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 constnied 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 commonly employed using conventional methodology by those skilled in tire art, such as, for example, the widely utilized molecular cloning methodologies described in Sambrook et al. , Molecular Cloning: A Laboratory Manual 2nd. edition (1 89) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N. Y. As appropriate, procedures involving the use of commercially available kits and reagents are generally carried out in accordance with manufacturer defined protocols and/or parameters unless otherwise noted.

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 die American Urological Association (AUA) system, stage CI - C2 disease under die 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 widi 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 die prostatic capsule, extends into the surgical margin, or invades the seminal vesicles.

"Altering the native glycosyiation pattern" is intended for purposes herein to mean deleting one or more carbohydrate moieties found in native sequence 213P1F11 (either by removing the underlying glycosyiation site or by deleting the glycosyiation by chemical and/or enzymatic means), and/or adding one or more glycosyiation sites that are not present in the native sequence 213P1F11. In addition, the phrase includes qualitative changes in die glycosyiation of the native proteins, involving a change in die nature and proportions of die various carbohydrate moieties present.

The term "analog" refers to a molecule which is structurally similar or sliares similar or corresponding attributes witii anodier molecule (e.g. a 213P1F11 -related protein). For example an analog of a 213P1F11 protein can be specifically bound by an antibody or T cell dial specifically binds to 213P1F11.

The term "antibody" is used in die broadest sense. Therefore an "antibody" can be naturally occurring or man-made such as monoclonal antibodies produced by conventional hybridoma teclmology. Anti-213P IFl 1 antibodies comprise monoclonal and polyclonal antibodies as well as fragments containing t e antigen-binding domain and/or one or more complementarity determining regions of these antibodies.

An "antibody fragment" is defined as at least a portion of d e variable region of die immunoglobulin molecule tliat binds to its target, i.e., die antigen-binding region. In one embodiment it specifically co\'ers single anti-213PlFll antibodies and clones tiiereof (including agonist, antagonist mid neutralizing antibodies) and anti-213PlFl l antibody compositions with polyepitopic specificity.

The term "codon optimized sequences" refers to nucleotide sequences dial 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 exon/intron splicing signals, elimination of transposon-like repeats and/or optimization of GC content in addition to codon optimization are referred to herein as mi "expression enhanced sequences." The term "cytotoxic agent" refers to a substance diat inhibits or prevents die expression activity of cells, function of cells and/or causes destruction of cells. The term is intended to include radioactive isotopes chemotlierapeutic 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 maytansinoids, yttrium, bismuth, ricin, ricin A-chain, doxorubicin, daunorubicin, taxol, etiiidium bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxy antiiracin dione, actinomycin, diphtheria toxin, Pseudomonas exotoxin (PE) A, PE40, abrin, abrin A chain, modeccin A chain, alpha-sarcin, gelonin, mitogellin, retstrictocin, phenomycin, enomycin, curicin, crotin, caliclieamicin, sapaonaria officinalis inliibitor, and glucocorticoid and other chemotlierapeutic agents, as well as radioisotopes such as At211 , 1131 , i125, Y90, Re186, Re188, Sm153, Bi212, P32 and radioactive isotopes of 11 Lu. Antibodies may also be conjugated to an anti-cancer pro-drug activating enzyme capable of converting die pro-drug to its active form.

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 class I or class II Major Histocompatibility Complex (MHC) protein (see, e.g. , Stites, et a/., IMMUNOLOGY, 8™ ED., Lange Publishing, Los Altos, CA ( 1994).

The terms "hybridize", "hybridizing", "hybridizes" and the like, used in the context of polynucleotides, are meant to refer to conventional hybridization conditions, preferably such as hybridization in 50% formamide/6XSSC/0. 1% SDS/100 ssDNA, in which temperatures for hybridization are above 37 degrees C and temperatures for washing in O. lXSSC/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 polynucleotide is said to be "isolated" when it is substantially separated from contaminant polynucleotides thai correspond or are complementary to genes oilier than the 213P1F11 genes or that encode polypeptides other than 213P1F11 gene product or fragments thereof. A skilled artisan can readily employ nucleic acid isolation procedures to obtain an isolated 213P1F11 polynucleotide. A protein is said to be "isolated," for example, when physical, mechanical or chemical methods are employed to remove the 213P1F1 1 proteins from cellular constituents that are normally associated with the protein. A skilled artisan can readily employ standard purification methods to obtain an isolated 213PIF11 protein. Alternatively, an isolated protein can be prepared by chemical means.

The tenn "mammal" 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, tire mammal 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 tire 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 metastatic 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 site for prostate cancer metastasis is bone. Prostate cancer bone metastases are often osteoblastic rather than osteolytic (i.e., resulting in net bone formation). Bone metastases are found most frequently in tire spine, followed by the femur, pelvis, rib cage, skull and humenis. Other common sites for metastasis include lymph nodes, lung, liver mid brain. Metastatic prostate cancer is typically diagnosed by open or laparoscopic pelvic lymphadenectomy, whole body radionuclide scans, skeletal radiography, and/or bone lesion biopsy.

The tenn "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the antibodies comprising tire population are identical except for possible naturally occurring mutations that are present in minor amounts. 12 A "motif, as in biological motif of a 213P 1 Fl 1 -related protein, refers to any pattern of amino acids forming part of tire primary sequence of a protein, that is associated with a particular function (e.g. protein-protein interaction, protein-DNA interaction, etc) or modification (e.g. that is phosphorylated, glycosylated or amidated), or localization (e.g. secretory sequence, nuclear localization sequence, etc.) or a sequence dial is correlated with being immunogenic, either Immorally or cellularly. A motif can be eidier contiguous or capable of being aligned to certain positions diat are generally correlated witii a certain function or property. In die context of HLA motifs, "motif refers to die pattern of residues in a peptide of defined lengtli, usually a peptide of from about 8 to about 13 amino acids for a class 1 HLA motif and from about 6 to about 25 amino acids for a class II HLA motif, which is recognized by a pardcular HLA molecule. Peptide motifs for HLA binding are typically different for each protein encoded by each human HLA allele and differ in die pattern of die primary and secondary anchor residues.

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

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

The term "polynucleotide" means a polymeric form of nucleotides of at least 10 bases or base pairs in lengtli, eidier ribonucleotides or deoxynucleotides 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 die art, diis term if often used interchangeably with "oligonucleotide". A polynucleotide can comprise a nucleotide sequence disclosed herein wherein diymidine (T), as shown for example in Figure 2, can also be uracil (U); diis definition pertains to die differences between die chemical structures of DNA and RNA, in particular the observation that one of the four major bases in R A is uracil (U) instead of thymidine (T).

The term "polypeptide" means a polymer of at least about 4, 5, 6, 7, or 8 amino acids. Throughout die specification, standard diree letter or single letter designations for amino acids are used. In die art, diis term is often used interchangeably with "peptide" or "protein".

An HLA "primary anchor residue" is an amino acid at a specific position along a peptide sequence which is understood to provide a contact point between die immunogenic peptide m d die HLA molecule. One to three, usually two, primary anchor residues widiin a peptide of defined length generally defines a "motif for an immunogenic peptide. These residues are understood to fit in close contact widi peptide binding groove of an HLA molecule, witii tiieir side chains buried in specific pockets of the binding groove. In one embodiment, for example, die primary anchor residues for an HLA class I molecule are located at position 2 (from die amino terminal position) and at the carboxyl terminal position of a 8, 9, 10, 11, or 12 residue peptide epitope in accordance widi die invention. In another embodiment, for example, die primary anchor residues of a peptide dial will bind an HLA class II molecule are spaced relative to each otiier, rather than to the termini of a peptide, where the peptide is generally of at least 9 amino acids in lengtli. The primary anchor positions for each motif and supennotif are set forth in Table IV. For example, analog peptides can be created by altering the presence or absence of particular residues in die primary and/or secondary anchor positions shown in Table IV. Such analogs are used to modulate die binding affinity and/or population coverage of a peptide comprising a particular HLA motif or supennotif. 13 A "recombinant" DNA or RN A molecule is a DNA or UNA molecule that has been subjected to molecular manipulation in vitro.

Non-limiting examples of small molecules include compounds that bind or interact with 213P1F11, ligands including hormones, neuropeptides, chemokines, odorants, phospholipids, and functional equivalents thereof that bind and preferably inhibit 213P 1F11 protein function. Such non-limiting small molecules preferably have a molecular weight of less than about 10 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, 213P1F1 1 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 ( ie relative temperature thai 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 ai, Current Protocols in Molecular Biology, Wiley Interscience Publishers, (1 95).

"Stringent conditions" or "high stringency conditions", as defined herein, are identified by, but not limited to, ttiose that: (1) employ low ionic strength and high temperature for washing, for example 0.015 M sodium chloride/0.0015 M sodium citrale/0.1% sodium dodecyl sulfate at 50°C; (2) employ during hybridization a denaturing agent, such as formamide, for example, 50% (v/v) formamide with 0.1% bovine serum albumin/0.1% Ficoll/0. i% polyvinylpyrrolidone/50 m sodium phosphate buffer at pH 6.5 with 750 niM sodium chloride, 75 mM sodium citrate at 42 °C; or (3) employ 50% formamide, 5 x SSC (0.75 M NaCl, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1 % sodium pyrophosphate, 5 Denhardt's solution, sonicated salmon sperm DNA (50 0.1% SDS, and 10% dextran sulfate at 42 °C, with washes at 42°C in 0.2 x SSC (sodium chloride/sodium, citrate) and 50% formamide at 55 °C, followed by a high-stringency wash consisting of 0.1 x SSC containing EDTA at 55 °C. "Moderately stringent conditions" are described by, but not limited to, those in Sambrook et ai, Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Press, 1989, and include the use of washing solution and hybridization conditions, (e.g., temperature, ionic strength and %SDS) less stringent than those described above. An example of moderalely stringent conditions is overnight incubation at 37°C in a solution comprising: 20% formamide, 5 x SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5 x Denhardt's solution, 10% dextran sulfate, and 20 mg/niL denatured sheared salmon sperm DNA, followed by washing the filters in 1 x SSC at about 37-50°C. The skilled artisan will recognize how to adjust the temperature, ionic strength, etc. as necessary to accommodate factors such as probe length and tire like.

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

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" (e.g., 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, e.g., 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 nucleic acids that encode such individual peptides or polypeptides, e.g. , a minigene that encodes a polyepitopic peptide. The "one or more peptides" can include any whole unit integer from 1 -242 or more, e.g., 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, 55, 60, 65, 70, 75, 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, or 242 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 class I peptides of the invention can be admixed with, or linked to, HLA class II peptides, to facilitate activation of both cytotoxic T lymphocytes and helper T lymphocytes. HLA vaccines can also comprise peptide-pulsed antigen presenting cells, e.g. , dendritic cells.

The term "variant" refers to a molecule that exliibits a variation from a described type or norm, such as a protein that has one or more different annuo acid residues in the corresponding position(s) of a specifically described protein (e.g. the 213P1F11 protein shown in Figure 2 or Figure 3. An analog is an example of a variant protein. Splice isoforms and single nucleotides polymorphisms (SNPs) are further examples of variants.

The "213PlFl l-related proteins" of tire 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 tire art. Fusion proteins that combine parts of different 213P1F11 proteins or fragments thereof, as well as fusion proteins of a 213P1F11 protein and a heterologous polypeptide are also included. Such 213P1F1 1 proteins are collectively referred to as the 213P1F11 -related proteins, the proteins of tire invention, or 213P1F11. The term "213P1F11-related protein" refers to a polypeptide fragment or a 213P1F1 I 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 titan 25 amino acids; or, at least 30, 35, 40, 45, 50, 55, 60, 65, 70, 80, 85, 90, 95, 100, 105, 110, 1 15, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 225, or 242 or more amino acids.

U.) 213P1F11 Polynucleotides One aspect of the invention provides polynucleotides corresponding, or complementary to all or part of a 213P1F11 gene, mRNA, and/or coding sequence, preferably in isolated form, including polynucleotides encoding a 213P1FI 1-related protein and fragments thereof, DNA, NA, DNA/RNA hybrid, and related molecules, polynucleotides or oligonucleotides complementary to a 213P 1F11 gene or mRNA sequence or a part thereof, and polynucleotides or oligonucleotides that hybridize to a 213P1F1 1 gene, mRNA, or to a 213P1F1 1 encoding polynucleotide (collectively, "213P1F11 polynucleotides"). In all instances when referred to in this section, T can also be U in Figure 2.

Embodiments of a 213P1 F11 polynucleotide include: a 213P1F11 polynucleotide having die sequence shown in Figure 2, the nucleotide sequence of 213P1F11 as shown in Figiire 2 wherein T is U; at least 10 contiguous nucleotides of a polynucleotide 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 213P1F1 1 nucleotides comprise, without limitation: (I) a polynucleotide 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 2 , from nucleotide residue number 404 through nucleotide residue number 1132, including tire stop codon, wherein T can also be U; (III) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in Figure 2B , from nucleotide residue number 404 through nucleotide residue number 1096, 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 nucleotide residue number 404 through nucleotide residue number 844, including the a stop codon, wherein T can also be U; (V) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in Figure 2D , from nucleotide residue number 1 through nucleotide residue number 966, including tire stop codon, wherein T can also be U; (VT) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in Figure 2E , from nucleotide residue number 404 through nucleotide residue number 1132, including the stop codon, wherein T can also be U; (VII) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in Figure 2F , from nucleotide residue number 404 through nucleotide residue number 1132, including the stop codon, wherein T can also be U; (VIII) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in Figure 2G , from nucleotide residue number 404 through nucleotide residue number 1 132, including the stop codon, wherein T can also be U; (IX) a polynucleotide comprising, consisting essentially of, or consisting of the sequence as shown in Figure 2H , from nucleotide residue number 404 through nucleotide residue number 1132, including the stop codon, wherein T can also be U; 16 (X) a polynucleotide that encodes a 213P 1 Fl 1 -related protein diat is at least 90% homologous to an entire amino acid sequence shown in Figure 2A-H ; (XI) a polynucleotide diat encodes a 213PlFl l-related protein dial is at least 90% identical to an entire amino acid sequence shown in Figure 2A-H ; (XII) a polynucleotide tliat encodes at least one peptide set forth in Tables V-X1X; (XIII) a polynucleotide tliat encodes a peptide region of at least 5 amino acids of a peptide of Figure 3A in any whole number increment up to 242 tliat includes an amino acid position having a value greater than 0.5 in the Hydrophilicity profile of Figure 5 A; or of Figure 3B in any whole number increment up to 230 tliat includes an amino acid position having a value greater dian 0.5 in the Hydrophilicity profile of Figure 5B; or of Figure 3C in any whole number increment up to 146 diat includes an amino acid position having a value greater than 0.5 in the Hydrophilicity profile of Figure 5C; or of Figure 3D in any whole number increment up to 321 thai includes an amino acid position having a value greater than 0.5 in die Hydrophilicity profile of Figure 5D; (XIV) a polynucleotide tliat encodes a peptide region of at least 5 amino acids of a pepdde of Figure 3A in any whole number increment up to 242 tliat includes an amino acid position having a value less titan 0.5 in the Hydropatliicity profile of Figure 6A; or of Figure 3B in any whole number increment up to 230 tliat includes an amino acid position having a value less dian 0.5 in the Hydropatliicity profile of Figure 6B; or of Figure 3C in any whole number increment up to 146 tliat includes an amino acid position having a value less than 0.5 in die Hydropatliicity profile of Figure 6C; or of Figure 3D in any whole number increment up to 321 tliat includes an amino acid position having a value less than 0.5 in the Hydropatliicity profile of Figure 6D; (XV) a polynucleotide diat encodes a peptide region of at least 5 amino acids of a peptide of Figure 3 A in any whole number increment up to 242 that includes an amino acid position having a value greater than 0.5 in the Percent Accessible Residues profile of Figure 7A; or of Figure 3B in any whole number increment up to 230 dial includes an amino acid position having a value greater dian 0.5 in d e Percent Accessible Residues profile of Figure 7B; or of Figure 3C in any whole number increment up to 146 diat includes an amino acid position having a value greater than 0.5 in die Percent Accessible Residues profile of Figure 7C; or of Figure 3D in any whole number increment up to 321 diat includes an amino acid position having a value greater than 0.5 in die Percent Accessible Residues profile of Figure 7D; (XVI) a polynucleotide diat encodes a peptide region of at least 5 amino acids of a peptide of Figure 3A in any whole number increment up to 242 that includes an amino acid position having a value greater dian 0.5 in die Average Flexibility profile of Figure 8 A; or of Figure 3B in any whole number increment up to 230 thai includes an amino acid position having a value greater than 0.5 in die Average Flexibility profile of Figure 8B; or of Figure 3C in any whole number increment up to 146 that includes an amino acid position having a value greater dian 0.5 in die Average Flexibility 17 profile of Figure8C; or of Figure 3D in any whole number increment up to 321 that includes an amino acid position having a value greater than 0.5 in the Average Flexibility profile of Figure 8D; (XVII) a polynucleotide that encodes a peptide region of at least 5 amino acids of a peptide of Figure 3 A in any whole number increment up to 242 that includes an amino acid position having a value greater than 0.5 in the Beta-turn profile of Figure 9A; or of Figure 3B in any whole number increment up to 230 that includes an amino acid position having a value greater than 0.5 in the Beta- turn profile of Figure 9B; or of Figure 3C in any whole number increment up to 146 that includes an amino acid position having a value greater than 0.5 in the Beta-turn profile of Figure9C; or of Figure 3D in any whole number increment up to 321 that includes an amino acid position having a value greater than 0.5 in the Beta-turn profile of Figure 9D; (XVIII) a polynucleotide that is fully complementary to a polynucleotide of any one of (I)-(XVTI).

(XIX) a peptide that is encoded by any of (l)-(XVIII); and (XXI) a polynucleotide of any of (I)-(XVIII) or peptide of (XIX) together with a pharmaceutical excipient and/or in a. human unit dose form.

As used herein, a range is understood to specifically disclose all whole unit positions thereo Typical embodiments of the invention disclosed herein include 213P1F11 polynucleotides that encode specific portions of 213P1F11 n R A sequences (aiid those which are complementary to such sequences) such as those that encode the proteins and/or fragments thereof, for example: (a) 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 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, 230, 235, 240, or 242 or more contiguous amino acids of 213P1F11. (b) 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, 70, 75, 80, 85, 90, 95, 100, 105, 1 10, 1 15, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, or 230 contiguous amino acids of variant 2; (c) 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, 70, 75, 80, 85, 90, 95, 100, 105, 1 10, 1 15, 120, 125, 130, 135, 140, 145, or 146 contiguous amino acids of variant 3 ; or (d) 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, or 146 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, or 321 contiguous amino acids of variant 4.

For example, representative embodiments of the invention disclosed herein include: polynucleotides and their encoded peptides themselves encoding about amino acid 1 to about amino acid 10 of the 213P1F11 protein shown in Figure 2 or Figure 3, polynucleotides encoding about amino acid 10 to about amino acid 20 of the 213P1F1 1 protein shown in Figure 2 or Figure 3, polynucleotides encoding about amino acid 20 to about amino acid 30 of the 213P1F11 protein shown in Figure 2 or Figure 3, polynucleotides encoding about 18 amino acid 30 to about amino acid 40 of the 213P1 F11 protein shown in Figure 2 or Figure 3, polynucleotides encoding about amino acid 40 to about amino acid 50 of the 213P1F11 protein shown in Figure 2 or Figure 3, polynucleotides encoding about amino acid 50 to about amino acid 60 of the 213P1F11 protein shown in Figure 2 or Figure 3, polynucleotides encoding about amino acid 60 to about amino acid 70 of the 213P1F11 protein shown in Figure 2 or Figure 3, polynucleotides encoding about ami o acid 70 to about amino acid 80 of the 213P1 F11 protein shown in Figure 2 or Figure 3, polynucleotides encoding about amino acid 80 to about amino acid 90 of the 213P1F11 protein shown in Figure 2 or Figure 3, polynucleotides encoding about amino acid 90 to about amino acid 100 of the 213P1F11 protein shown in Figure 2 or Figure 3, in increments of about 10 amino acids, ending at the carboxyl terminal amino acid set forth in Figure 2 or Figure 3.

Accordingly polynucleotides encoding portions of the amino acid sequence (of about 10 amino acids), of amino acids 100 through the carboxyl terminal amino acid of the 213P1F11 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 213P1F11 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 40 or 50 etc.) of the 213P1F11 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 213P1F11 sequence as shown in Figure 2.

Additional illustrative embodiments of the invention disclosed herein include 213P1F11 polynucleotide fragments encoding one or more of the biological motifs contained within a 213P1 F11 protein "or variant" sequence, including one or more of the motif-bearing subsequences of a 213 P1 F1 1 protein "or variant" set forth in Tables V-X1X.

Note that to determine the star-ting position of any peptide set forth in Tables V-XiX (collectively HLA Peptide Tables) respective to its parental protein, e.g., variant 1, variant 2, etc., reference is made to three factors: (lie par ticular variant, the length of the peptide in an HLA Peptide Table, and the Search Peptides in Table XXIX. 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 XXIX. Accordingly if a Search Peptide begins at position "X", one must add the value "X - 1" to each position in Tables V-XIX 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 is parental molecule, one must add 150 -1, i.e., 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 V-XIX collectively, or an oligonucleotide Uiat encodes the HLA peptide. Another embodiment of the invention comprises an HLA peptide that occurs at least once in Tables V-XVlll and at least once in table XIX, or an oligonucleotide that encodes the HLA peptide.

Another embodiment of the invention is antibody epitopes which comprise a peptide regions, or an oligonucleotide encoding die peptide region, that has one two, three, four, or five of the following characteristics: 1 i) a peptide region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to die full lengdi of that protein in Figure 3, tliat includes an amino acid position having 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 die Hydrophilicity profde of Figure 5; ii) a peptide region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to die full length of that protein in Figure 3, that includes an amino acid 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 peptide region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to die full lengdt of diat protein in Figure 3, diat 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 die Percent Accessible Residues profile of Figure 7; iv) a pepdde region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to die full length of that protein in Figure 3, tliat includes an amino acid position having a value equal to or greater dian 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in die Average Flexibility profile of Figure 8; or v) a peptide region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to die full length of that protein in Figure 3 , dial includes an amino acid posi tion havi ng 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 die Beta-turn profile of Figure 9.

In ahodier embodiment, typical polynucleotide fragments of die invention encode one or more of die regions of 2 I3P1F1 I 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 213P1F11 protein or variant N-glycosylation sites, cAMP and cGMP -dependent protein kinase phosphorylation sites, casein kinase II phosphorylation sites or N-n yristoylatioii site and amidation sites.

II.A.) Uses of 213P1F11 Polynucleotides Il.A.l.) Monitoring of Genetic Abnormalities The polynucleotides of the preceding paragraphs have a number of different specific uses. The human 213P1F11 gene maps to the chromosomal location set fordi in ie Example entitled "Chromosomal Mapping of 2I 3P1FH ." For example, because the 213P1F11 gene maps to dus chromosome, polynucleotides that encode different regions of the 213P1F11 proteins are used to characterize cytogenetic abnormalities of tliis chromosomal locale, such as abnormalities diat are identified as being associated with various cancers. In certain genes, a variety of chromosomal abnormalities including rearrangements have been identified as frequent cytogenetic abnormalities in a number of different cancers (see e.g. Krajinovic et al. , Mutat. Res. 382(3-4): 81-83 (1998); Johansson et al., Blood 86(10): 3905-3914 (1995) mid Finger et al., P.N.A.S. 85(23): 9158-9162 (1988)). Thus, polynucleotides encoding specific regions of the 213P1F11 proteins provide new tools that can be used to delineate, widi greater precision dian previously possible, cytogenetic abnormalities in die chromosomal region dial encodes 213P1F1 1 diat may contribute to the malignant phenotype. In diis context, diese polynucleotides satisfy a need in die art for expanding die sensitivity of chromosomal screening in order to identify more subtle and less common chromosomal abnormalities (see e.g. Evans er a/., Am. J. Obstet. Gynecol 171(4): 1055-1057 (1994)).

Furthermore, as 213P1F11 was shown to be highly expressed in bladder aiid olher cancers, 213P1F1 1 polynucleotides are used in methods. assessing die status of 213 P1FI 1 gene products in normal versus cancerous tissues. Typically, polynucleotides that encode specific regions of the 213P1F1 1 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 213P 1F11 gene, suc as regions containing one or more motifs. Exemplary assays include both RT-PCR assays as well as single-strand conformation polymorphism (SSCP) analysis (see, e.g., Marrogi et al, J. Cutan. Pathol. 26(8): 369-378 (1999), both of which utilize polynucleotides encoding specific regions of a protein to examine these regions within the protein.

II.A.2.) Antisense Embodiments Other specifically contemplated nucleic acid related embodiments of the invention disclosed herein are genomic DNA, cDNAs, ribozymes, and antisense molecules, as well as nucleic acid molecules based on an alternative backbone, or including alternative bases, whether derived from natural sources or synthesized, and include molecules capable of inhibiting the RNA or protein expression of 213 P 1 Fl 1. For example, antisense molecules can be RN As or other molecules, including peptide nucleic acids (PNAs) or non-nucleic acid . molecules such as phosphorotliioate derivatives, that specifically bind DNA or RNA in a base pair-dependent manner. A skilled artisan can readily obtain these classes of nucleic acid molecules using the 213P1F11 polynucleotides and polynucleotide 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 oligonucleotides are complementary to their intracellular targets, e.g., 213P 1F11. See for example, Jack Cohen, Oligodeoxynucleotides, Antisense Inliibitors of Gene Expression, CRC Press, 1989; and Synthesis 1 : 1 -5 (1988). The 213P1F1 1 antisense oligonucleotides of the present invention include derivatives such as S-oligonucleotides (phosphorotliioate derivatives or S-oligos, see, Jack Cohen, supra), which exhibit enhanced cancer cell growth inhibitory action. S-oligos (nucleoside phosphorothioates) are isoelectronic analogs of an oligonucleotide (O-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 vvitli 3H- l ,2-benzodithiol-3-one-l , l-dioxide, which is a sulfur transfer reagent. See, e.g., Iyer, R. P. et al , J. Org. Chem. 55:4693-4698 (1990); and Iyer, R. P. et al , J. Am. Chem. Soc. 1 12: 1253-1254 ( 1990). Additional 213P1F1 1 antisense oligonucleotides of the present invention include niorpholino antisense oligonucleotides known in the art (see, e.g., Partridge et al, 1996, Antisense & Nucleic Acid Drug Development 6: 169-175).

The 213P1F11 antisense oligonucleotides 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 213P 1F1 1 genomic sequence or (lie corresponding niRNA. Absolute complementarity is not required, although high degrees of complementarity are preferred. Use of an oligonucleotide complementary to this region allows for die selective hybridization to 213P1F11 mRNA and not to niRNA specifying other regulatory subunits of protein kinase. In one embodiment, 213P1F11 antisense oligonucleotides of the present invention are 15 to 30-nier fragments of the antisense DNA molecule that have a sequence that 21 hybridizes to 213P1F1 1 mRNA. Optionally, 213P1F11 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 213P1F1 1. Alternatively, the antisense molecules are modified to employ ribozymes in the iimibition of 213P1F11 expression, see, e.g., L. A. Couture & D. T. Stinchcomb; Trends Genet 12: 510-515 (1996). 1I.A.3.) Primers and Primer Pairs Further specific embodiments of this nucleotides of the invention include primers and primer pairs, which allow the specific amplification of polynucleotides 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 213P1F11 polynucleotide in a sample and as a means for detecting a cell expressing a 213P1F11 protein.

Examples of such probes include polypeptides comprising all or part of the human 213P1F11 cDNA sequence shown in Figure 2. Examples of primer pairs capable of specifically amplifying 213P1F11 iiiRNAs 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 213P1F11 mRNA.

The 213P1F 1 1 polynucleotides of the invention are useful for a variety of purposes, including but not limited to their use as probes and primers for tire amplification and/or detection of (lie 213P1F11 gene(s), lnRNA(s), or fragments thereof; as reagents for the diagnosis and/or prognosis of prostate cancer and oilier cancers; as coding sequences capable of directing (lie expression of 213P1 F11 polypeptides; as tools for modulating or inhibiting the expression of the 213P1F11 gene(s) and/or translation of the 213P1F11 ' transcript(s); and as therapeutic agents.

The present invention includes the use of any probe as described herein to identify and isolate a 213P1 F11 or 213P1F11 related nucleic acid sequence from a naturally occurring source, such as humans or other mammals, as well as the isolated nucleic acid sequence per se, which would comprise all or most of the sequences found in the probe used.■ Π.Α.4.) Isolation of 213PlFl l-Encoding Nucleic Acid Molecules The 213P1F1 1 cDNA sequences described herein enable the isolation of other polynucleotides encoding 213P1 F11 gene product(s), as well as tlie isolation of polynucleotides encoding 213P1F11 gene product homoJogs, alternatively spliced isofonns, allelic variants, and mutant forms of a 213P1F11 gene product as well as polynucleotides that encode analogs of 213PlFl l-related proteins. Various molecular cloning methods that can be employed to isolate full length cDNAs encoding a 213P1F11 gene are well known (see, for example, Sambrook, J. et al. , Molecular Cloning: A Laboratory Manual, 2d edition, Cold Spring Harbor Press, New York, 1989; Current Protocols in Molecular Biology. Ausubel et al, Eds., Wiley mid Sons, 1995). For example, lambda phage cloning methodologies can be conveniently employed, using commercially available cloning systems (e.g., Lambda ZAP Express, Stratagene). Phage clones containing 213P1F11 gene cDNAs can be identified by probing with a labeled 213P1F11 cDNA or a fragment thereof. For example, in one embodiment, a 213P1F11 cDNA (e.g., Figure 2) or a portion thereof can be synthesized and used as a probe to retrieve overlapping and full-length cDNAs corresponding to a 213P1F11 gene. A 213P1F11 gene itself can be isolated 22 by screening genomic DNA libraries, bacterial artificial chromosome libraries (BACs), yeast artificial chromosome libraries (YACs), and the like, with 213P1F11 DNA probes or primers.

H.A.5.) Recombinant Nucleic Acid Molecules and Host-Vector Systems The invention also provides recombinant DNA or RNA molecules containing a 213P1F 11 polynucleotide, a fragment, analog or homologue thereof, including but not limited to phages, plasmids, 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 molecules. Methods for generating such molecules are well known (see, for example, Sambrook et al. , 1989, supra).

The invention further provides a host-vector system comprising a recombinant DNA molecule containing a 213P1F1 1 polynucleotide, fragment, analog or homologue thereof within a suitable prokaryotic or eukaryotic host cell. Examples of suitable eukaryotic host cells include a yeast cell, a plant cell, or an animal cell, such as a mammalian cell or an insect cell (e.g., a baculovirus-infectible cell such as mi SP or HighFive cell). Examples of suitable mammalian cells include various prostate cancer cell lines such as DU145 and TsuPrl , other transfectable or transducible prostate cancer cell lines, primary cells (PrEC), as well as a number of mammalian cells routinely used for the expression of recombinant proteins (e.g., COS, CHO, 293, 293T cells). More particularly, a polynucleotide comprising the coding sequence of 213P1F11 or a fragment, analog or homolog thereof can be used to generate 213P1F11 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 213P1F11 proteins or fragments thereof are available, see for example, Sambrook et al., 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 pSRottkneo (Muller et al., 1991, MCB 11 : 1785). Using these expression vectors, 213P1F11 can be expressed in several prostate cancer and non-prostate cell lines, including for example 293, 293T, rat-1, NLH 3T3 and TsuPrl. The host-vector systems of the invention are useful for the production of a 213P1F1 1 protein or fr agment thereof. Such host-vector systems can be employed to study the functional properties of 213P1F1 1 and 213P1F1 1 mutations or analogs.

Recombinant human 213P1F1 1 protein or an analog or homolog or fragment thereof can be produced by mammalian cells transfected with a construct encoding a 213P 1F1 1 -related nucleotide. For example, 293T cells can be transfected with an expression plasmid encoding 213P1F11 or fragment, analog or homolog thereof, a 213P1F11 -related protein is expressed in the 293T cells, and the recombinant 213P1F1 1 protein is isolated using standard purification methods (e.g., affinity purification using anti-213P1 F1 1 antibodies). In another embodiment, a 213P1F1 1 coding sequence is subcloned into the retroviral vector pSRaMS Vtkneo and used to infect various mammalian cell lines, such as NIH 3T3, TsuPrl, 293 and rat-1 in order to establish 213P1F11 expressing cell lines. Various oilier expression systems well known in the art can also be employed. Expression constructs encoding a leader peptide joined in frame to a 213P1F1 1 coding sequence can be used for the generation of a secreted form of recombinant 213P1 F1 1 protein.

As discussed herein, redundancy in the genetic code permits variation in 213 P I F 11 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 (i.e., codons having a usage frequency of less than about 20% in luiown 23. 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 located at the World Wide Web .dna.affrc.go.jp/~nalcamura/codon.html.

Additional sequence modifications are known to enhance protein expression in a cellular host. These include elimination of sequences encoding spurious polyadenylation signals, exon/intron 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 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 ozak, Mol. Cell Biol., 9:5073-5080 (1989). Skilled artisans lurderstand that the general rule that eukaryotic ribosomes initiate translation exclusively at the 5' proximal AUG codon is abrogated only under rare conditions (see, e.g., Kozak PNAS 92(7): 2662-2666, (1995) and Kozak NAR 15(20): 8125-8148 (1987)).

III.) 213P1F11-i-elated Proteins Another aspect of the present invention provides 213P1F1 1 -related proteins. Specific embodiments of 213P 1 F1 1 proteins comprise a polypeptide having all or part of the amino acid sequence of human 213P1F1 1 as shown in Figure 2 or Figure 3. Alternatively, embodiments of 213P1 F11 proteins comprise variant, homolog or analog polypeptides that have alterations in the amino acid sequence of 213P1F11 shown * in Figure 2 or Figure 3.

In general, naturally occurring allelic variants of human 213P1 F1 1 share a high degree of structural identity and homology (e.g., 90% or more homology). Typically, allelic variants of a 213P1F11 protein contain conservative amino acid substitutions within the 213P1F11 sequences described herein or contain a substitution of an amino acid from a corresponding position in a homologue of 213P1F11. One class of 213P1F11 allelic variants are proteins that share a liigh degree of homology with at least a small region of a particular 213P1F11 amino acid sequence, but further contain a radical departure from the sequence, such as a non-conservative substitution, tnmcation, insertion or frame sluft. 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 acid 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 isoleucine (I), valine (V), and leucine (L) for any other of these hydrophobic amino acids; aspartic acid (D) for glutamic acid (E) and vice versa; glutamine (Q) for asparagine (N) and vice versa; and serine (S) for threonine (T) and vice versa. Other substitutions can also be considered conservative, depending on die environment of the particular amino acid and its role in the three-dimensional structure of the protein. For example, glycine (G) and alanine (A) can frequently be interchangeable, as can alanine (A) and valine (V). Methionine (M), which is relatively hydrophobic, can frequently be interchanged with leucine and isoleucine, and sometimes widi valine. Lysine (K) and arginine (R) are frequently 24 interchangeable in locations in which the significant feature of the amino acid residue is its charge and the differing p 's of these two amino acid residues are not 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 ai, PNAS 1992 Vol 89 10915-10919; Lei et ai, J Biol Chem 1995 May 19; 270(20): 11882-6).

Embodiments of the invention disclosed herein include a wide variety of art-accepted variants or analogs of 213P 1F 1 1 proteins such as polypeptides having amino acid insertions, deletions and substitutions. 213P 1 Fl 1 variants can be made using methods known in the art such as site-directed mutagenesis, alanine scanning, and PCR mutagenesis. Site-directed mutagenesis (Carter et ai , Nucl. Acids Res., 73:4331 (1986); Zoller et ai, Nucl. Acids Res., 70:6487 (1987)), cassette mutagenesis (Wells et ai , Gene, 34:315 (1985)), restriction selection mutagenesis (Wells et ai , Philos. Trans. R. Soc. London SerA, 317:415 (1986)) or other known techniques can be performed on the cloned DNA to produce the 213P1F11 variant DNA.

Scanning amino acid 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 scaiming amino acids are relatively small, neutral amino acids. Such amino acids include alanine, glycine, serine, and cysteine. Alanine is typically a preferred scanning amino acid among this group because it eliminates tire side-chain beyond the beta-carbon mid 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 mid exposed positions (Creighton, The Proteins, (W.H. Freeman & Co., N.Y.); Chothia, J. Mol. Biol., 150: 1 (1976)). If alanine substitution does not yield adequate amounts of variant, mi isosteric amino acid can be used.

As defined herein, 213P1F11 variants, analogs or homologs, have the distinguishing attribute of having at least one epitope that is "cross reactive" witii a 213P1F1 1 protein having mi amino acid sequence of Figure 3. As used in tins sentence, "cross reactive" means tliat mi mitibody or T cell diat specifically binds to a 213P1F11 variant also specifically binds to a 213P1F11 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 mitibody or T cell uiat specifically binds to the starting 213P1F11 protein. Those skilled in the art understand tliat antibodies that recognize proteins bind to epitopes of varying size, and a grouping of die order of about four or five amino acids, contiguous or not, is regarded as a typical number of amino acids in a minimal epitope. See, e.g., Nair et ai, J. Immunol 2000 165(12): 6949-6955; Hebbes et ai, Mol Immunol (1989) 26(9):865-73; Schwartz et ai , J Immunol ( 1985) 135(4):2598-608.

Odier classes of 213P 1F1 1-related protein variants share 70%, 75%, 80%, 85% or 90% or more similarity with mi amino acid sequence of Figure 3, or a fragment thereof. Anodier specific class of 213P 1F1 1 protein variants or analogs comprise one or more of the 213P1F11 biological motifs described herein or presently known in the art. Thus, encompassed by the present invention are analogs of 213P1F11 fragments (nucleic or amino acid) 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 die nucleic or amino acid sequences of Figure 2 or Figure 3.

As discussed herein, embodiments of die claimed invention include polypeptides containing less than the full amino acid sequence of a 213P 1FU protein shown in Figure 2 or Figure 3. For example, .25 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 acids of a 213P1F11 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 213P1F11 protein shown in Figure 2 or Figure 3, polypeptides consisting of about amino acid 10 to about amino acid 20 of a 213P1F11 protein shown in Figure 2 or Figure 3„ polypeptides consisting of about amino acid 20 to about amino acid 30 of a 213P1F11 protein shown in Figure 2 or Figure 3, polypeptides consisting' of about amino acid 30 to about amino acid 40 of a 213P1F11 protein shown in Figure 2 or Figure 3, polypeptides consisting of about amino acid 40 to about amino acid 50 of a 213P1F1 1 protein shown in Figure 2 or Figure 3, polypeptides consisting of about amino acid 50 to about amino acid 60 of a 213P1F11 protein shown in Figure 2 or Figure 3, polypeptides consisting of about amino acid 60 to about amino acid 70 of a 213P 1F11 protein shown in Figure 2 or Figure 3, polypeptides consisting of about amino acid 70 to about amino acid 80 of a 213P 1F11 protein shown in Figure 2 or Figure 3, polypeptides consisting of about amino acid 80 to about amino acid 90 of a 213P1F1 1 protein shown in Figure 2 or Figure 3, polypeptides consisting of about amino acid 90 to about amino acid 100 of a 213P1F1 1 protein shown in Figure 2 or Figure 3, etc. throughout die entirety of a 213P1F11 amino acid sequence. Moreover, polypeptides consisting of about amino acid 1 (or 20 or 30 or 40 etc.) to about amino acid 20, (or 130, or 140 or 150 etc.) of a 213P1F11 protein shown in Figure 2 or Figure 3 are embodiments of tlie invention. It is to be appreciated that tlie starting and slopping positions in tins paragraph refer to tlie specified position as well as that position plus or minus 5 residues. 213PlFl l -related proteins are generated using standard peptide synthesis technology or using chemical clea vage methods well known in tlie art. Alternatively, recombinant methods can be used to generate nucleic acid molecules that encode a 213P1F 11-related protein. In one embodiment, nucleic acid molecules provide a means to generate defined fragments of a 213P1F1 1 protein (or variants, homologs or analogs thereof).

III. A.) Motif-bearint; Protein Embodiments Additional illustrative embodiments of the invention disclosed herein include 213P 1F1 1 polypeptides comprising the amino acid residues of one or more of the biological motifs contained within a 213P1 Fl 1 polypeptide sequence set forth in Figure 2 or Figure 3. Various motifs are known in tlie art, and a protein can be evaluated for tlie presence of such motifs by a number of publicly a vailable Internet sites (see, e.g., URL addresses located at tlie World Wide Web: pfam.wustl.edu/; http://searclilauncher.bcin.tmc.edu/seq-searcli struc-predict.htinl; psort.ims.u-tokyo.ac.jp/; .cbs.dtu.dk/; .ebi.ac.uk/interpro/scan.html; .expasy.cli/tools/scnpsitl .html; Epimatrix™ and Epiiner™, Brown University, .brovvn.edii/Researcli/TO-HIV_Lab/epiinatrix/epiniatrix.hliiil; and BlMAS, biinas.dcrt.nili.gov/.).

Motif bearing subsequences of all 213P1F1 1 variant proteins are set forth and identified in Tables V- XIX.

Table XX sets forth several frequently occurring motifs based on pfam searches (see URL address pfam.wustl.edu ). The columns of Table XX list (1) motif name abbreviation, (2) percent identity found amongst tlie different member of tlie motif family, (3) motif name or description and (4) most common function; location information is included if the motif is relevant for location.

Polypeptides comprising one or more of the 213PIF11 motifs discussed above are useful 'in elucidating the specific characteristics of a malignant phenotype in view of the observation that tlie 213P1F1 1 26 motifs discussed above are associated with growth dysregulation and because 213P1F1 1 is overexpressed in certain cancers (See, e.g., Table I). 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 al. ; Lab Invest., 78(2): 165-174 (1998); Gaiddon et al. , Endocrinology 136(10): 4331 -4338 (1995); Hall et al. , Nucleic Acids Research 24(6): 1119-1 126 (1996); Pelerziel et al., Oncogene 18(46): 6322-6329 (1999) and O'Brian, Oncol. Rep. 5(2): 305-309 (1998)). Moreover, both glycosylation and myristoylation are protein modifications also associated with cancer and cancer progression (see e.g. Dennis et al. , Biochem. Biophys. Acta 1473(l):21-34 (1999); Raju et at. , Exp. Cell Res. 235(1): 145-154 (1997)). Amidation is another protein modification also associated witli cancer and cancer progression (see e.g. Treston ei al., J. Natl. Cancer Inst. Monogr. (13): 169-175 (1992)).

In another embodiment, proteins of the invention comprise one or more of the iininunoreactive epitopes identified in accordance with art-accepted methods, such as the peptides set forth in Tables V-XIX. CTL epitopes can be determined using specific algorithms to identify peptides witliin a 213P1F11 protein that are capable of optimally binding to specified HLA alleles (e.g., Table IV; Epimatrix™ and Epimer™, Brown University, URL located at the World Wide Web . brown. edii/Researcli TB-HIV_Lab/epimatrix/epiniatrix.htiiil; mid BIMAS, URL biinas.dcrt.nili.gov/.) Moreover, processes for identifying peptides that have sufficient binding affinity for HLA molecules and which are correlated witli 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 epitopes, are well known in lire art, and are carried out without undue experimentation either in vitro or in vivo.

Also known in the art are principles for creating analogs of such epitopes in order to modulate immunogenicity. For example, one begins witli an epitope that bears a CTL or HTL motif (see, e.g., 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, one can substitute out a deleterious residue in favor of any other residue, such as a preferred residue as defined in Table IV; substitute a less-preferred residue witli a prefen ed residue as defined in Table IV; or substitute an originally-occurring preferred residue witli another preferred residue as defined in Table IV. S bstitutions can occur at primary anchor positions or at other positions in a peptide; see, e.g., Table IV.

A variety of references reflect t e art regarding the identification and generation of epitopes in a protein of interest as well as analogs thereof. See, for example, WO 9733602 to Chesnut et al. ; Sette, lmmunogenetics 1999 50(3-4): 201-212; Sette et al., J. Immunol. 2001 166(2): 1389-1397;' Sidney et al. , Hum. Immunol. 1997 58(1): 12-20; Kondo et al, lmmunogenetics 1997 45(4): 249-258; Sidney et al., J. Immunol. 1996 157(8): 3480-90; and Falk et al., Nature 351 : 290-6 (1991); Hunt et al. , Science 255: 1261-3 (1992); Parker et al., J. Immunol. 149:3580-7 (1992); Parker et al., J. Immunol. 152: 163-75 (1994)); Rast er al , 1994 152(8): 3904-12; Borras-Cuesta et al , Hum. Immunol. 2000 61(3): 266-278; Alexander et al., J. Immunol. 2000 164(3); 164(3): 1625-1633; Alexander et al. , PMID: 7895164, UI: 95202582; O' Sullivan et al., J. Immunol. 1991 147(8): 2663-2669; Alexander et al., Immunity 1994 1 (9): 751-761 and Alexander et al. , Immunol. Res. 1998 18(2): 79-92. 27 Related embodiments of the invention include polypeptides comprising combinations of the different motifs set fort h in Table XXI, and/or, one or more of the predicted CTL epitopes of Tables V-X1X, and/or, one or more of the T cell binding motifs known in tire art. Preferred embodiments contain no insertions, deletions or substitutions either within the motifs or the intervening sequences of the polypeptides. In addition, embodiments which include a number of either N-terminal and/or C-terininal amino acid residues on either side of these motifs may be desirable (to, for example, include a greater portion of the polypeptide arclutecture 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 acid residues. 213P1F11-related proteins are embodied in many forms, preferably in isolated form. A purified 213P1F1 1 protein molecule will be substantially free of other proteins or molecules that impair the binding of 213P1 F1 1 to antibody, T cell or other ligand. The nature and degree of isolation and purification will depend on the intended use. Embodiments of a 213P1F11-related proteins include purified 213P1F1 1 -related proteins and functional, soluble 213PlFl l-related proteins. In one embodiment, a functional, soluble 213P1F1 1 protein or fragment thereof retains the ability to be bound by antibody, T cell or other ligand.

The invention also provides 213P1F11 proteins comprising biologically active fragments of a 213P1F1 1 amino acid sequence shown in Figure 2 or Figure 3. Such proteins exhibit properties of the starting 213P1F1 1 protein, such as the ability to elicit the generation of antibodies that specifically bind an epitope associated with the starting 213P1 F11 protein; lo be bound by such antibodies; to elicit the activation of HTL or CTL; and/or, to be recognized by HTL or CTL that also specifically bind to the starting protein. 213P1F11-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, Garnier-Robson, yte-Doolittle, Eisenberg, Karplus-Schultz or Jameson-Wolf analysis, or on the basis of imnumogenicity. Fragments that contain such stractures are particularly useful in generating sub unit-specific anti-213PlFl l antibodies, or T cells or in identifying cellular factors that bind to 213P1F11. For example, hydrophilicity profiles can be generated, and immunogenic peptide fragments identified, using the method of Hopp, T.P. and Woods, K.R., 1981 , Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828. Hydropathicity profiles can be generated, and immunogenic peptide fragments identified, using the method of Kyte, J. and Doolittle, R.F., 1982, J. Mol. Biol. 157: 105-132. Percent (%) Accessible Residues profiles can be generated, and immunogenic peptide fragments identified, using the method of Janin J., 1979, Nature 277:491-492. Average Flexibility profiles can be generated, and immunogenic peptide fragments identified, using the method of Bhaskaran R., Ponnuswamy P. ., 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, G., Roux B., 1987, Protein Engineering 1 :289-294.

CTL epitopes can be determined using specific algorithms to identify peptides within a 213PIF1 1 protein that are capable of optimally binding to specified HLA alleles (e.g., by usin die SYFPEITHI site at World Wide Web URL syfpeimi.bmi-heidelberg.com/; the listings in Table IV(A)-(E); Epimatrix™ and Epimer™, Brown University, located at the World Wide Web (.brown.edu/Research/TB-HlV_Lab/epimalrix/epimatrix.htinl); and B1MAS, URL bimas.dcrt.mh.gov/). Illustrating this, peptide epitopes from 213P1F11 that are presented in the context of human MHC Class 1 molecules, e.g., HLA-A1, A2, A3, 28 Al 1, A24, B7 and B35 were predicted (Tables V-XIX). Specifically, the complete amino acid sequence of tlie 213P1F11 protein and relevant portions of other variants, i.e., for HLA Class I predictions 9 flanking redisues on either side of a point mutation, and for HLA Class II predictions 14 flanking residues on either side of a point mutation, were entered into tl e HLA Peptide Motif Search algoritlun found in tlie Bioinformatics and Molecular Analysis Section (B1MAS) web site listed above; in addition to the site SYFPEITHI, at URL syfpeitlii.bmi-heidelberg.com/.

The HLA peptide motif search algoritlun was developed by Dr. Ken Parker based on binding of specific peptide sequences in tlie groove of HLA Class I molecules, in particular HLA-A2 (see, e.g., Falk et al , Nature 351 : 290-6 (1991); Hunt et al, Science 255: 1261-3 (1992); Parker et at., J. Immunol. 149:3580-7 (1992); Parker et al , J. Immunol. 152 : 163-75 (1994)). This algoritlun allows location mid ranking of 8-mer, 9- mer (also refered to as "nonamer"), and 10-nier (also refered to as "decamer") peptides from a complete protein sequence for predicted binding to HLA-A2 as well as numerous other HLA Class I molecules. Many HLA class 1 binding peptides are 8-, 9-, 10 or 11 -mers. For example, for Class I HLA-A2, tlie epitopes preferably contain a leucine (L) or methionine (M) at position 2 and a valine (V) or leucine (L) at tlie C-terminus (see, e.g., Parker et al , j. Immunol. 149:3580-7 (1992)). Selected results of 213P1F11 predicted binding peptides are shown in Tables V-XIX herein. In Tables V-XIX, tlie selected candidates, 9-mers and - mers, mid 15-mers for each family member are shown along with their location, tlie amino acid sequence of each specific peptide, and an estimated binding score. The binding score corresponds to tlie estimated half time of dissociation of complexes containing tlie peptide at 37°C at pH 6.5. Peptides vvitli lhe lughest binding score are predicted to be tlie most tightly bound to HLA Class I on tlie cell surface for tlie greatest period of time and thus represent tlie best immunogenic targets for T-cell recognition.

Actual binding of peptides to an HLA allele can be evaluated by stabilization of HLA expression on tlie antigen-processing defective cell line T2 (see, e.g., Xue et al , Prostate 30:73-8 (1997) mid Peshwa et al , Proslate 36: 129-38 (1998)). Imniunogenicity of specific peptides can be evaluated in vitro by stimulation of CD8+ cytotoxic T lymphocytes (CTL) in die presence of antigen presenting cells such as dendritic cells.

It is to be appreciated that every epitope predicted by the BIMAS site, Epimer™ and Epimatrix™ sites, or specified by the HLA class I or class II motifs available in tlie art or which become part of the art such as set forth in Table IV (or determined using World Wide Web site URL syfpeithi.bmi-heidelberg.com/, or BIMAS, binias.dcrt.nih.gov/) are to be "applied" to a 213P1F11 protein in accordance vvidi tlie.invention. As used in this context "applied" means that a 213P1F11 protein is evaluated, e.g., visually or by computer-based patterns finding methods, as appreciated by those of skill in tlie relevant art. Every subsequence of a 213P1F11 protein of 8, 9, 10, or 11 amino acid residues that bears mi HLA Class I motif, or a subsequence of 9 or more amino acid residues that bear mi HLA Class II motif are within tlie scope of the invention.

III.B.) Expression of 213PlFll-reliiteil Proteins In an embodiment described in the examples that follow, 213P1F1 1 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 213 P 1 F11 with a C-terminal 6XHis and MYC tag (pcDNA3.1/mycHIS, Invitrogen or Tag5, GeiiHunter Corporation, Nashville TN). The Tag5 vector provides mi IgGK secretion signal that can be used to facilitate (lie production of a secreted 213P 1F11 protein in transfected cells. The 29 secreted HIS-tagged 213P1F11 in the culture media ca be purified, e.g., using a nickel column using standard techniques. ni.C.) Modifications of 213Pt Fl t-rclated Proteins Modifications of 213P1F1 1-related proteins such as covalenl modifications are included witliin die scope of this invention. One type of covalenl modification includes reacting targeted amino acid residues of a 213P1FH polypeptide with an organic derivalizing agent dial is capable of reacting with selected side chains or tlie N- or C- terminal residues of a 213P1F11 protein. Another type of covalenl modification of a 213P1F1 1 polypeptide included within the scope of tins invention comprises altering the native glycosylation pattern of a protein of die invention. Anodier type of covalent modification of 213P1F11 comprises linking a 213P1F1 1 polypeptide to one of a variety of nonproteinaceous polymers, e.g., 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 213PlFl l-related proteins of die present invention can also be modified to form a chimeric molecule comprising 213P1F11 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 anodier tumor-associated antigen or fragment thereof. Alternatively, a protein in accordance with the invention can comprise a fusion of fragments of a 213P IF11 sequence (amino or nucleic acid) such that a molecule is created that is not, through its length, directly homologous to die amino or nucleic acid sequences shown in Figure 2 or Figure 3. Such a chimeric molecule can comprise multiples of the same subsequence of 213P1F11. A chimeric molecule can comprise a fusion of a 213P1F11 -related protein wiUi 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 die amino- or carboxyl- terminus of a213PlFl 1 protein. In an alternative embodiment, die chimeric molecule can comprise a fusion of a 213P1F11-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 "immiinoadhesin"), such a fusion could be to die Fc region of an IgG molecule. The Ig fusions preferably include die substitution of a soluble (transmembrane domain deleted or inactivated) form of a 213P1F11 polypeptide in place of at least one variable region wid in an g molecule. In a preferred embodiment, the immunoglobulin fusion includes die l inge, CH2 and CH3, or d e lunge, CHI, CH2 and CH3 regions of an lgGI molecule. For die production of immunoglobulin fusions see, e.g., U.S. Patent No. 5,428,130 issued June 27, 1995.

HID.) Uses of 213P1 F11-related Proteins The proteins of d e invention have a number of different specific uses. As 213P1F11 is highly expressed in prostate and other cancers, 213PlFl l-related proteins are used in methods diat assess die status of 213P1 F11 gene products in normal versus cancerous tissues, Uiereby elucidating the malignant phenotype. Typically, polypeptides from specific regions of a 213P1F11 protein are used to assess die presence of perturbations (such as deletions, insertions, point mutations etc.) in diose regions (such as regions containing one or more motifs). Exemplary assays utilize antibodies or T cells targeting 213P1F11 -related proteins comprising die amino acid residues of one or more of the biological motifs contained widiin a 213P1F1 1 polypeptide sequence in order to evaluate die characteristics of this region in normal versus cancerous tissues or to elicit an immune response to die epitope. Alternatively, 213P1F11-related proteins that contain d e amino acid residues of one or more of the biological motifs in a 213P1F11 protein are used to screen for factors that interact with that region of 213P1F11. 213P1F11 protein fragments/subsequences are particularly useful in generating and characterizing domain-specific antibodies (e.g., antibodies recognizing an extracellular or intracellular epitope of a 213P1F11 protein), for identifying agents or cellular factors that bind to 213PIF11 or a particular structural domain diereof, and in various tlierapeutic and diagnostic contexts, including but not limited to diagnostic assays, cancer vaccines and methods of preparing such vaccines.

Proteins encoded by the 213P1F11 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 mat bind to a 213P1F11 gene product. Antibodies raised against a 213P1F1 1 protein or fragment thereof are useful in diagnostic and prognostic assays, and imaging methodologies in the management of human cancers characterized by expression of 213P1F11 protein, such as those listed in Table I. Such antibodies can be expressed intracellularly and used in methods of treating patients with such cancers. S213P1F11-related nucleic acids or proteins are also used in generating HTL or CTL responses.

Various immunological assays useful for the detection of 213P1FI 1 proteins are used, including but not limited to various types of radioimmunoassays, enzyme-linked immunosorbent assays (ELISA), enzyme-linked immunofluorescenl assays (ELIFA), iimiiunocytochemical methods, and the like. Antibodies can be labeled and used as immunological imaging reagents capable of detecting 213P1F11-expressing cells (e.g., in radioscintigraphic imaging methods). 213P1 Fl 1 proteins are also particularly useful in generating cancer vaccines, as further described herein.

IV.) 213P1F11 Antibodies Another aspect of (lie invention provides antibodies that bind to 213P1F11-related proteins. Preferred antibodies specifically bind to a 213P1F11 -related protein and do not bind (or bind wealdy) to peptides or proteins that are not 213P1F11-related proteins. For example, antibodies that bind 213P1F11 can bind 213P1F11-related proteins such as the homologs or analogs thereof. 2 13P1 F11 antibodies of the invention are particularly useful in cancer (see, e.g., Table I) diagnostic and prognostic assays, and imaging methodologies. Similarly, such antibodies are useful in the treatment, diagnosis, and/or prognosis of other cancers, to the extent 213P1F11 is also expressed or overexpressed in diese other cancers. Moreover, intracellularly expressed antibodies (e.g., single chain antibodies) are therapeutically useful in treating cancers in wluch tire expression of 213P1F1 1 is involved, such as advanced or metastatic prostate cancers.

The invention also provides various immunological assays useful for die detection and quantification of 213P1F11 and mutant 213PlFll-related proteins. Such assays can comprise one or more 213P1F11 antibodies capable of recognizing and binding a 213P1F11-related protein, as appropriate. These assays are performed within various immunological assay formats well known in the art, including but not limited to various types of radioimmunoassays, enzyme-linked immunosorbent assays (ELISA), enzyme-linked immunofluorescent assays (ELIFA), and the like. 31 Immunological non-antibody assays of the invention also comprise T cell inimunogenicity 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 213P1F11 are also provided by tlie invention, including but not limited to radioscintigraphic imaging methods using labeled 213P1F11 antibodies. Such assays are clinically useful in tlie detection, monitoring, and prognosis of 213P1F11 expressing cancers such as prostate cancer. 213P1F11 antibodies are also used in methods for purifying a 213PlFl l-related protein and for isolating 213P1F11 homologues arid related molecules. For example, a method of purifying a 213P1F11 -related protein comprises incubating a 213P1F11 antibody, which has been coupled to a solid matrix, with a lysate or other solution containing a 213P1F11 -related protein under conditions that permit tlie 213P1 Fl l antibody to bind to tlie 213P1F 1 1 -related protein; washing tlie solid matrix to eliminate impurities; and eluting tl e 213P1F11 -related protein from tlie coupled antibody. Other uses of 213P1F1 1 antibodies in accordance with the invention include generating anti-idiotypic antibodies that mimic a 213P1F11 protein.

Various methods for the preparation of antibodies are well known in tlie art. For example, antibodies can be prepared by immunizing a suitable mammalian host using a 213PlFl l-related protein, peptide, or fragment, in isolated or iminunoconjugated 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 213P1F11 can also be used, such as a 213P1F11 GST-fusion protein. In a particular embodiment, a GST fusion J protein comprising all or most of lire amino acid sequence of Figure 2 or Figure 3 is produced, the used as an immunogen to generate appropriate antibodies. In another embodiment, a 213PlFl l-related protein is synthesized and used as an immunogen.

In addition, naked DNA immunization techniques known in tlie art are used (with or without purified 213P 1F1 1 -related protein or 213P1F11 expressing cells) to generate an immune response to tlie encoded immunogen (for review, see Donnelly et ai , 1997, Ann. Rev. Immunol. 15: 617-648).

The amino acid sequence of a 213P1F11 protein as shown in Figure 2 or Figure 3 can be analyzed to select specific regions of tlie 213P1F11 protein for generating antibodies. For example, hydrophobicity and liydroplulicity analyses of a 213P1F11 amino acid sequence are used to identify hydropliilic regions in tlie 213P1F11 structure. Regions of a 213P1F11 proteiii that show immunogenic structure, as well as other regions mid domains, can readily be identified using various other methods known in tlie art, such as Chou-Fasman, Garnier-Robson, Kyte-Doolittle, Eisenberg, Karplus-Schultz or Jameson- Wolf analysis. Hydrophilicity profiles can be generated using die method of Hopp, T.P. and Woods, K.R., 1981, Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828. Hydropathicity profiles can be generated using tlie method of Kyte, J. and Doolittle, R.F., 1982, J. Mol. Biol. 157: 105-132. Percent (%) Accessible Residues profiles can be generated using tl e method of Janin J., 1979, Nature 277:491-492. Average Flexibility profiles can be generated using tlie method of B askaran R., Ponnuswamy P.K., 1988, Int. J. Pept. Protein Res. 32:242-255. Beta-turn profiles can be generated using tlie method of Deleage, G., Roux B., 1987, Protein Engineering 1 :289-294. Thus, each region identified by any of these programs or methods is witliin the scope of the present invention. Methods for tl e generation of 213P1F11 antibodies are further illustrated by way of tlie examples provided herein. Methods , for preparing a protein or polypeptide for use as an inuniuiogen are well known in the art. Also well known in- the art are methods for preparing immunogenic conjugates of a protein with a carrier, such as BSA, KLH or other 32 carrier protein. In some circumstances, direct conjugation using, for example, carbodiimide reagents are used; in oilier instances linking reagents such as diose supplied by Pierce Chemical Co., Rockford, IL, are effective. Administration of a 213P IF 11 imnuinogen is often conducted by injection over a suitable time period and with, use of a suitable adjuvant, as is understood in the art. During the immunization schedule, titers of antibodies can be taken to determine adequacy of antibody formation. 213P1F11 monoclonal antibodies can be produced by various means well known in the art. For example, immortalized cell lines that secrete a desired monoclonal antibody are prepared using tire standard hybridoma teclmology of Koliler and ilstein 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 wliich tire antigen is a 213P1F11-related protein. When tire appropriate immortalized cell culture is identified, the cells can be expanded and antibodies produced either from in vitro 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 213P1F11 protein can also be produced in tire context of chimeric or complementarity determining region (CDR) grafted antibodies of multiple species origin. Humanized or human 213P1F1 1 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 et al., 1988, Nature 332: 323-327; Verhoeyen et al., 1988, Science 239: 1534-1536). See also, Carter et al. , 1993, Proc. Natl. Acad. Sci. USA 89: 4285 mid Sims et al., 1993, J. Immunol. 151 : 2296.

Methods for producing fully human monoclonal antibodies include phage display and transgenic methods (for review, see Vaughan et ai, 1998, Nature Biotechnology 16: 535-539). Fully human 213P1F11 monoclonal antibodies can be generated using cloning technologies employing large human Ig gene combinatorial libraries (i.e., phage display) (Griffiths and Hoogenboom, Building an in vitro immune system: human antibodies from phage display libraries. In: Protein Engineering of Antibody Molecules for Prophylactic and Therapeutic Applications in Man, Clark, M. (Ed.), Nottingham Academic, pp 45-64 (1993); Burton and Barbas, Human Antibodies from combinatorial libraries. Id., pp 65-82). Fully human 213P1F11 monoclonal antibodies can also be produced using transgenic mice engineered to contain human immunoglobulin gene loci as described in PCT Patent Application W098/24893, Kucherlapati and Jakobovits et al. , published December 3, 1997 (see also, Jakobovits, 1998, Exp. Opin. Invest. Drugs 7(4): 607-614; U. S. patents 6, 162,963 issued 19 December 2000; 6, 150,584 issued 12 November 2000; and, 6,114598 issued 5 September 2000). This method avoids the in vitro manipulation required with phage display technology and efficiently produces high affinity authentic human antibodies.

Reactivity of 213P 1F11 antibodies with a 213P1F11 -related protein can be established by a number of well known means, including Western blot, irnmunoprecipitation, ELISA, and FACS analyses using, as appropriate, 213P1F1 1-related proteins, 213PlFl l-expressing cells or extracts tliereof. A 213P1F11 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 radioisotope, a fluorescent compound, a bioluminescent compound, chemiluminescent compound, a metal chelator or mi enzyme. Further, bi-specific antibodies specific for two or more 213P1F1 1 epitopes are generated using methods generally known in the art. 33 Homodimeric antibodies can also be generated by cross-linking techniques known in the art (e.g., Wolff et al. , Cancer Res. 53 : 2560-2565).

V.) 213P1F11 Cellular Immune Responses The mechanism by which T cells recognize antigens lias been delineated. Efficacious peptide epitope vaccine compositions of the invention induce a therapeutic or prophylactic immune responses in very broad segments of the world-wide population. For an understanding of the value and efficacy of compositions of the invention that induce cellular immune responses, a brief review of immunology-related technology is provided.

A complex of an HLA molecule and a peptidic antigen acts as the ligand recognized by HLA-restricted T cells (Buns, S. et al. , Cell 47: 1071, 1986; Babbitt, B. P. et al., Nature 317:359, 1985; Townsend, A. and Bodmer, Η., Αηηιι. Rev. Immunol. 7:601, 1989; Germain, R. N., /1W/7M. Rev. Immunol. 11 :403, 1993). Through the study of single amino acid substituted antigen analogs and the sequencing of endogeiioiisly bound, naturally processed peptides, critical residues that correspond to motifs required for specific binding to HLA antigen molecules have been identified and are set fortli in Table IV (see also, e.g. , Soutliwood, et al. , J. Immunol. 160:3363, 1998; Rammensee, et al. , Immunogenetics 41 : 178, 1995; Ranm ensee et al. , SYFPE1THI, access via World Wide Web at URL syfpeithi.bmi-lieidelberg.com7; Sette, A. and Sidney, J. Curr. Opin. Immunol. 10:478, 1998; Engelhard, V. H., Cun: Opi . Immunol. 6: 13, 1994; Sette, A. and Grey, H. M, Curr. Opin. Immunol. 4:79, 1992; Sinigaglia, F. and Hammer, J. Curr. Biol. 6:52, 1994; Ruppert et al., Cell 74:929-93.7, 1993; Kondo et al. , J. Immunol. 155:4307-4312, 1995; Sidney et al. , 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 crystallograpliic 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 ligands; these residues in turn determine the HLA binding capacity of the peptides in which they are present. (See, e.g., Madden, D.R. Annu. Rev. Immunol. 13:587, 1995; Smith, et al., Immunity 4:203, 1996; Fremont et al., Immunity 8:305, 1998; Stem et al., Structure 2:245, 1994; Jones, E.Y. .Curr. Opin. Immunol. 9:75, 1997; Brown, J. H. et al., Nature 364:33, 1993; Gno, H. C. et al., Proc. Natl. Acad. Sci. USA 90:8053, 1993; Guo, H. C. et al., Nature 360:364, 1992; Silver, M. L. et al. , Nature 360:367, 1992; Matsumura, M. et al. , Science 257:927, 1992; Madden et al., Cell 70: 1035, 1992; Fremont, D. H. el al. , Science 257:919, 1992; Saper, M. A. , Bjorkman, P. J. and Wiley, D. C, J. Mot. Biol. 219:277, 1991.) Accordingly, the definition of class' I and class 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 confirmat ory work can be performed to select, amongst these vacci ne candidates, epitopes with preferred characteristics in terms of population coverage, and/or imniunogenicity.

Various strategies can be utilized to evaluate cellular imniunogenicity, including: 34 1) Evaluation of primary T cell cultures from normal individuals (see, e.g. , Wentworth, P. A. et al, Mot. Immunol. 32:603, 1995; Celis, E. et al. , Pro Natl. Acad. Sci. USA 91 :2105, 1994; Tsai, V. et at., J. Immunol. 158: 1796, 1997; awashiina, 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 weeks. T cells specific for tire peptide become activated during this time and are detected using, e.g. , a lymphokine- or -^ Cr-release assay involving peptide sensitized target cells. 2) Immunization of HLA transgenic mice- (see, e.g. , Wentworth, P. A. et l. , J. Immunol. 26:97, 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 subculaneously to HLA transgenic mice. Several weeks following immunization, spienocytes are removed and cultured in vitro in lire presence of test peptide for approximately one week. Peptide-specific T cells are detected using, e.g., a -^Cr-release assay involving peptide sensitized target cells and target cells expressing endogenously generated antigen. 3) Demonstration of recall T cell responses from immune individuals who have been eidier effectively vaccinated and/or from chronically ill patients (see, e.g. , Rehermann, B. et al., J. Exp. Med. 181 : 1047, 1995; Doolan, D. L. et al. , Immunity 1:91, 1997; Bertoni, R. et al., J. Clin. Invest. 100:503, 1997; Threlkeld, S. C. et al., J. Immunol. 159: 1648, 1997; Diepolder, H. M. et al. , J. Virol. 71 :6011, 1997).

Accordingly, recall responses are detected by culturing PBL from subjects that have been exposed to the antigen due to disease and thus have generated an immune response "naturally", or from patients who were vaccinated against the antigen. PBL from subjects are cultured in vitro for 1 -2 weeks in lire presence of test peptide plus antigen presenting cells (APC) to allow activation of "memory" T cells, as compared to "naive" T cells. Al the end of the culture period, T cell activity is detected using assays including -^Cr release involving peptide-sensitized targets, T cell proliferation, or lymphokine release.

VI.) 213P1F11 Transgenic Animals Nucleic acids that encode a 213PlFl l-related protein can also be used to generate either transgenic animals or "knock out" animals that, in turn, are useful in tire development and screening of therapeutically useful reagents. In accordance with established techniques, cDNA encoding 213P1F11 can be used to clone genomic DNA that encodes 213P1F11. The cloned genomic sequences can tlien.be used to generate transgenic animals containing cells that express DNA that encode 213P1F11. Methods for generating transgenic animals, particularly animals such as mice or rats, have become conventional in the art mid 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 213PIF1 i transgene incorporation with tissue-specific enhancers.

Transgenic animals that include a copy of a transgene encoding 213P1F11 can be used to examine the effect of increased expression of DNA thai encodes 213P1F1 1. Such animals can be used as tester animals for reagents thought to confer protection from, for example, pathological conditions associated 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.

Alternatively, non-human hoinologues of 213P1F11 can be used to construct a 213P1F1 1 "knock out" animal that has a defective or altered gene encoding 213P1F11 as a result of homologous recombination between t e endogenous gene encoding 213P1F11 and altered genomic DNA encoding 213P1F1 1 introduced into an embryonic cell of the animal. For example, cDNA that encodes 213P1F11 can be used to clone genomic DNA encoding 213P1F11 in accordance with established techniques. A portion of the genomic DNA encoding 213P1F11 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, e.g., Thomas and Capecchi, Cell, 51:503 (1987) for a description of homologous recombination vectors). The vector is introduced into an embryonic stem cell line (e.g., by electroporation) and cells in which the introduced DNA has homologoiisly reconibined with the endogenous DNA are selected (see, e.g., Li et al., Cell, 69:915 (1992)). The selected cells are then injected into a blastocyst of an animal (e.g., a mouse or rat) to form aggregation chimeras (see, e.g., Bradley, in Teratocarcinomas and Embryonic Stern Cells: A Practical Approach, E. J. Robertson, ed. (IRL, Oxford, 1987), pp. 113-152). A chimeric embryo can (hen be implanted into a suitable pseudopregnant female foster, animal, and the embryo brought to term to create a "knock out" animal. Progeny harboring (lie homologoiisly reconibined DNA in dieir germ cells can be identified by standard techniques and used to breed animals in which all cells of the animal contain the homologoiisly reconibined DNA. Knock out animals can be characterized, for example, for their ability to defend against certain pathological conditions or for their development of pathological conditions due to absence of a 213P1F11 polypeptide.

VII) Methods for the Detection of 213P1F11 Another aspect of the present invention relates to methods for detecting 213P1F11 polynucleotides and 213PlFl l-related proteins, as well as methods for identifying a cell that expresses 213P1F11. The expression profile of 213P1F11 makes it a diagnostic marker for metastasized disease. Accordingly, the status of 213P1 F1 1 gene products provides information useful for predicting a variety offactors including susceptibility to advanced stage disease, rate of progression, and/or tumor aggressiveness. As discussed in detail herein, the status of 213P1F1 1 gene products in patient samples can be analyzed by a variety protocols that are well .known in the. art including immunohistochemicaJ analysis, the variety of Northern blotting techniques including in situ hybridization, RT-PCR analysis (for example on laser capture micro-dissected samples), Western blot analysis and tissue array analysis.

More particularly, (lie invention provides assays for the detection of 213P1F11 polynucleotides in a biological sample, such as serum, bone, prostate, and other tissues, urine, semen, cell preparations, and the like. Detectable 213P1F11 polynucleotides include, for example, a 213P1F11 gene or fragment thereof, 213P1F11 niRNA, alternative splice variant 213P1F11 niRNAs, and recombinant DNA or RNA molecules that contain a 213P1F11 polynucleotide. A number of methods for amplifying and/or detecting the presence of 213P1F11 polynucleotides are well known in (lie art and can be employed in (lie practice of this aspect of the invention.

In one embodiment, a method for detecting a 213P1F1 1 niRNA in a biological sample comprises producing cD A from the sample by reverse transcription using al least one primer; amplifying the cDNA so 36 produced vising a 213P1F11 polynucleotides as sense and antisense primers to amplify 213P1F11 cDNAs therein; and detecting die presence of the amplified 213P1F11 cDNA. Optionally, the sequence of the amplified 213P1F11 cDNA can be determined.

In another embodiment, a method of detecting a 213P1F11 gene in a biological sample comprises first isolating genomic DNA from the sample; amplifying d e isolated genomic DNA using 213P IF 11 polynucleotides as sense and antisense primers; mid detecting the presence of the amplified 213P1F1 1 gene. Any number of appropriate sense and antisense probe combinations can be designed from a 213P1F1 1 nucleotide sequence (see, e.g., Figure 2) and used for this purpose.

The invention also provides assays for detecting die presence of a 213PIF11 protein in a tissue or other biological sample such as semm, semen, bone, prostate, urine, cell preparations, and die like. Mediods for detecting a 213Pl Fl l-related protein are also well known and include, for example, immunoprecipitation, inunuiiohistochemical analysis, Western blot analysis, molecular binding assays, EL1S A, ELIFA and die like. For example, a method of detecting the presence of a 213P1F1 l-rela(ed protein in a biological sample comprises first contacting the sample with a 213P1F1 1 antibody, a 213P1F11 -reactive fragment diereof, or a recombinant protein containing mi antigen binding region of a 213P1F11 antibody; and then detecting Uie binding of 213P1F1 1 -related protein in die sample.

Mediods for identifying a cell dial expresses 213P1F11 are also within the scope of die invention. In one embodiment, mi assay for identifying a cell that expresses a 213P1F11 gene comprises detecting die presence of 213P1F1 1 niRNA in die cell. Methods for the detection of particular niRNAs in cells are well known and include, for example, hybridization assays using complementary DNA probes (such as in situ hybridization using labeled 213P1F11 riboprobes, Nordiern blot and related techniques) mid various nucleic acid miiplification assays (such as RT-PCR using complementary primers specific for 2 I3P1F11, and odier amplification type detection mediods, such as, for example, branched DNA, SISBA, TMA mid die like). Alternatively, mi assay for identifying a cell that expresses a 213P1F11 gene comprises detecting the presence of 213PlFl l-related protein in die cell or secreted by the cell. Various mediods for the detection of proteins are well known in die art and are employed for the detection of 213P1F11-related proteins and cells that express 213P1F1 1-related proteins. 213P1F11 expression analysis is also useful as a tool for identifying mid evaluating agents that modulate 213P1F11 gene expression. For example, 213P1F11 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 dial inhibits 213P1F11 expression or over-expression in cancer cells is of tlierapeiitic value. For example, such an agent can be identified by using a screen tiiat quantifies 213P1F1 1 expression by RT-PCR, nucleic acid hybridization or antibody binding.

Vin.) Methods for Monitoring the Status of 213P1F11-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 mid tiien cancerous states (see, e.g., Alers et al., Lab Invest. 77(5): 437-438 (1997) mid Isaacs et al., Cancer Surv. 23 : 19-32 (1995)). In diis context, examining a biological sample for evidence of dysregulated cell growth (such as aberrant 213P1F11 expression in cancers) allows for early detection of such aberrant physiology, before a pathologic state such as cancer has progressed to a stage Uiat therapeutic options me more limited and or die prognosis is 37 worse. In such examinations, the status of 213P1F11 in a biological sample of interest can be compared, for example, to the status of 213P1F1 1 in a corresponding normal sample (e.g. a sample from that individual or alternatively another individual that is not affected by a pathology). An alteration in the status of 213P1F11 in the biological sample (as compared to the normal sample) provides evidence of dysregulated cellular growth. In addition to using a biological sample' thai 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 niR A expression (see, e.g., Grever el al , J. Comp. Neurol. 1996 Dec 9; 376(2): 306-14 and U. S. Patent No. 5,837,501) to compare 213P1 11 status in a sample.

The term "status" in tius context is used according to its art accepted meaning and refers to the condition or state of a gene and its products. Typically, skilled artisans use a number of parameters to evaluate die condition or state of a gene and its products. These include, but are not limited to tire location of expressed gene products (including the location of 213P 1F1 1 expressing cells) as well as the level, and biological activity of expressed gene products (such as 213P1F11 mRNA, polynucleotides and polypeptides). Typically, an alteration in the status of 213P1F11 comprises a change in the location of 213P1F11 and/or 213P1 F1 1 expressing cells and/or an increase in 213P1F11 mRNA and/or protein expression. 213P1F11 status in a sample can be analyzed by a number of means well known in the art, including witiiout limitation, inuininomslochemical analysis, in situ hybridization, RT-PCR analysis on laser capture micro-dissected samples, Western blot analysis, and tissue array analysis. Typical protocols for evaluating die status of a 213P1F11 gene and gene products are found, for example in Ausubel et al. eds., 1995, Current Protocols In Molecular Biology, Units 2 (Northern Blotting), 4 (Southern Blotting), 15 (Immunoblotting) and 18 (PCR Analysis). Thus, the status of 213P1F1 1 in a biological sample is evaluated by various mediods utilized by skilled artisans including, but not limited to genomic Southern, analysis (to examine, for example perturbations in a 213P1F1 1 gene), Northern analysis and/or PCR analysis of 213P1F11 mRNA (to examine, for example alterations in the polynucleotide sequences or expression levels of 213P1F1 1 niRNAs), and, Western and/or immunoliistochemical analysis (to examine, for example alterations in polypeptide sequences, alterations in polypeptide localization vvitliin a sample, alterations in expression levels of 213P1F11 proteins and/or associations of 213P1FI 1 proteins with polypeptide binding partners). Detectable 213P IF 1 1 polynucleotides include, for example, a 213P1F11 gene or fragment Uiereof, 213P1FI 1 mRNA, alternative splice variants, 213P1F11 niRNAs, and recombinant DNA or RNA molecules containing a 213P1F11 polynucleotide.

The expression profile of 213P1F11 makes it a diagnostic marker for local and/or metastasized disease, and provides information on die growth or oncogenic potential of a biological sample. In particular, the status of 213P1F11 provides infonnation useful for predicting susceptibility to particular disease stages, progression, and/or tumor aggressiveness. The invention provides mediods m d assays for determining 213P1F1 1 status and diagnosing cancers that express 213P1F11 , such as cancers of die tissues listed in Table I. For example, because 213P1F11 mRNA is so highly expressed in prostate and otiier cancers relative to normal prostate tissue, assays dial evaluate the levels of 213P1F1 1 mRNA transcripts or proteins in a biological sample can be used to diagnose a disease associated with 213P1F11 dysregulation, and can provide prognostic infonnation useful in defining appropriate dierapeutic options.

The expression status of 213P1F11 provides infonnation including die presence, stage and location of dysplastic, precancerous and cancerous cells, predicting susceptibility to various stages of disease, and/or for 38 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 213P1F1 1 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 213P1F11 in a biological sample can be examined by a number of well-known procedures in the art. For example, the status of 213P1F11 in a biological sample taken from a specific location in the body can be examined by evaluating tire sample for tire presence or absence of 213P1F11 expressing cells (e.g. those that express 213P1F11 mRNAs or proteins). This examination can provide evidence of dysregulated cellular growth, for example, when 213P1F1 l-expressing cells are found in a biological sample that does not normally contain such cells (such as a lymph node), becanse such alterations in die status of 213P1F11 in a biological sample are often associated with dysregulated cellular growth. Specifically, one indicator of dysregulated cellular growth is die metastases of cancer cells from an organ of origin (such as die prostate) to a different area of the body (such as a lymph node). In diis context, evidence of dysregulated cellular growth is important for example because occult lymph node metastases can be delected in a substantial proportion of patients with prostate cancer, and such metastases are associated vvidi known predictors of disease progression (see, e.g., Murphy et al., Prostate 42(4): 315-317 (2000);Su et al. , Semin. Surg. Oncol. 18(1): 17-28 (2000) and Freeman et al., J Urol 1995 Aug 154(2 Pt l):474-8).

In one aspect, the invention provides methods for monitoring 213P1F1 1 gene products by determining the status of 213P1F11 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 213P1F11 gene products in a corresponding normal sample. The presence of aberrant 213P1F11 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, die invention provides assays useful in determining the presence of cancer in an individual, comprising detecting a significant increase in 213P1F11 mRNA or protein expression in a test cell or tissue sample relative to expression levels in die corresponding normal cell or tissue. The presence of 213P 1F11 mRNA can, for example, be evaluated in tissues including but not limited to Uiose listed in Table I. The presence of significant 213P 1F11 expression in any of these tissues is useful to indicate the emergence, presence and/or severity of a cancer, since the corresponding normal tissues do not express 213P1F1 1 mRNA or express it at lower levels.

In a related embodiment, 213P1F1 1 status is determined at lire protein level rather than at the nucleic acid level. For example, such a method comprises determining the level of 213P1F11 protein expressed by cells in a test tissue sample and comparing the level so determined to the level of 213P1F11 expressed in a corresponding normal sample. In one embodiment, tire presence of 213P1F11 protein is evaluated, for example, using imrnunohislocheniical methods. 213P1F11 antibodies or binding partners capable of detecting 213P1F11 protein expression are used in a variety of assay formats well known in the art for tliis purpose.

In a further embodiment, one can evaluate the status of 213P1F11 nucleotide and amino acid sequences in a biological sample in order to identify perturbations in the structure of these molecules. These perturbations can include insertions, deletions, substitutions and the like. Such evaluations are useful because perturbations in 39 tlie nucleotide and amino acid sequences are observed in a large number of proteins associated with a growth dysregulaled phenotype (see, e.g., Marrogi et al, 1999, J. Cutan. Pathol. 26(8):369-378). For example, a mutation in tlie sequence of 213P1F1 1 may be indicative of tlie presence or promotion of a tumor. Such assays therefore have diagnostic and predictive value where a mutation in 213P1F11 indicates a potential loss of function or increase in tumor growth.

A wide variety of assays for observing perturbations in nucleotide and amino acid sequences are well known in the art. For example, tl e size and structure of nucleic acid or amino acid sequences of 2I3P1F11 gene products are observed by the Northern, Southern, Western, PCR and DNA sequencing protocols discussed herein. In addition, other methods for observing perturbations in nucleotide and amino acid sequences such as single strand conformation polymorphism analysis are well laiown in tlie art (see, e.g., U.S. Patent Nos. 5,382,510 issued 7 September 1999, and 5,952,170 issued 17 January 1995).

Additionally, one can examine Uie nietliylalion status of a 213P1F11 gene in a biological sample.

Aberrant demethylation and/or hypermelhylalion 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 hypermefhylation of tlie 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 tlie 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 70% of cases of high-grade prostatic intraepithelial neoplasia (PIN) (Brooks et al, Cancer Epidemiol. Biomarkers Prev., 1998, 7:531 -536). In another example, expression of the LAGE-1 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 nietliylalion status of a gene are well known in tlie art. For example, one can utilize, in Southern hybridization approaches, meUiylatioii-sensitive restriction enzymes that cannot cleave sequences that contain methylated CpG sites to assess tlie niethylation status of CpG islands. In addition, MSP (niethylation specific PCR) can rapidly profile tlie niethylation status of all tlie 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 uumethylated cytosines to uracil) followed by amplification using primers specific for methylated versus unmefhylated DNA. Protocols involving methylation interference can also be found for example in Current Protocols In Molecular- Biology, Unit 12, Frederick M. Aiisubel et al. eds., 1995.

Gene amplification is an additional method for assessing tlie status of 213P1F11. Gene amplification is measured in a sample directly, for example, by conventional Southern blotting or Northern blotting to quantilaie tlie transcription of mRNA (Thomas, 1980, Proc. Natl. Acad. Sci. USA, 77:5201 -5205), dot blotting (DNA analysis), or in situ hybridization, using an appropriately labeled probe, based on tlie sequences provided herein. Alternatively, antibodies are employed that recognize specific duplexes, including DNA duplexes, RNA duplexes, and DNA-RNA hybrid duplexes or DNA-protein duplexes. The antibodies in turn are labeled and tlie assay carried out where tlie duplex is bound to a surface, so that upon tlie formation of duplex on tlie surface, tlie presence of antibody bound to the duplex can be detected.

Biopsied tissue or peripheral blood can be conveniently assayed for tlie presence of cancer cells using for example, Northern, dot blot or RT-PCR analysis to detect 213P1FU expression. The presence of RT-PCR 40 amplifiable 213PlFl l mRNA provides an indication of lire presence of cancer. RT-PCR assays are well known in Hie art. RT-PCR detection assays for minor cells in peripheral blood are currently being evaluated for use in die diagnosis mid management of a number of human solid tumors. In die prostate cancer field, tiiese include RT-PCR assays for the detection of cells expressing PSA and PSM (Verkaik et al, 1 97, Urol. Res. 25:373-384; Ghossein ei al. , 1995, J. Clin, Oncol. 13: 1195-2000; Heston ei al., 1995, Clin. Chem. 41 :1687-1688).

A further aspect of the invention is an assessment of the susceptibility that an individual has for developing cancer. In one embodiment, a mediod for predicting susceptibility to cancer comprises detecting 213P1F11 mRNA or 213P1F11 protein in a tissue sample, its presence indicating susceptibility to cancer, wherein d e degree of 213P1F11 mRNA expression correlates to die degree of susceptibility. In a specific embodiment, die presence of 213P1F11 in prostate or odier tissue is examined, witii die presence of 213P1F11 in die sample providing an indication of prostate cancer susceptibility (or die emergence or existence of a prostate tumor). Similarly, one can evaluate d e integrity 213P1F11 nucleotide and amino acid sequences in a biological sample, in order to identify perturbations in d e structure of these molecules such as insertions, deletions, substitutions and the like. The presence of one or more perturbations in 213P1F11 gene products in die sample is an indication of cancer susceptibility (or die emergence or existence of a tumor).

The invention also comprises methods for gauging tumor aggressiveness. In one embodiment, a metiiod for gauging aggressiveness of a tumor comprises determining die level of 213P1F11 mRNA or 213P1F11 protein expressed by tumor cells, comparing die level so determined to die level of 213P1F11 mRNA or 213P1F11 protein expressed in a corresponding nonnal tissue taken from the same individual or a normal tissue reference sample, wherein die degree of 213P1F11 mRNA or 213P1F11 protein expression in die tumor sample relative to the nonnal sample indicates die degree of aggressiveness. In a specific embodiment, aggressiveness of a tumor is evaluated by deteniiining die extent to Av ich 213P1F11 is expressed in die tumor cells, widi higher expression levels indicating more aggressive tumors. Anodier embodiment is the evaluation of die integrity of 2I3P1F1 1 nucleotide and amino acid sequences in a biological sample, in order to identify' perturbations in die structure of diese molecules such as insertions, deletions, substitutions and die like. The presence of one or more perturbations indicates more aggressive tumors.

Another embodiment of the invention is directed to metiiods for obsendng die progression of a malignancy in mi individual over time. In one embodiment, metiiods for observing die progression of malignancy in an individual over time comprise determining die level of 213P1F11 mRNA or 213P1F11 protein expressed by cells in a sample of die tumor, comparing d e level so deleniiined to die level of 213P1 F11 mRNA or 213P 1F11 protein expressed in an equivalent tissue sample taken from die same individual at a different time, wherein the degree of 213P1F11 mRNA or 213P1F11 protein expression in die tumor sample over time provides infoniiation on die progression of die cancer. In a specific embodiment, die progression of a cancer is evaluated by determining 213P1F11 expression in d e tumor cells over time, where increased expression over time indicates a progression of die cancer. Also, one can evaluate die integrity 213P1F11 nucleotide and amino acid sequences in a biological sample in order to identify perturbations in die structure of these molecules such as insertions, deletions, substitutions m d die like, where d e presence of one or more perturbations indicates a progression of (lie cancer.

The above diagnostic approaches can be combined witii any one of a wide variety of prognostic and diagnostic protocols known in die art. For example, anotiier embodiment of the invention is directed to metiiods 41 for observing a coincidence between the expression of 213P1FU gene and 213P1F11 gene products (or perturbations in 213P1F11 gene and 213P JF1 J gene products) and a factor that is associated with malignancy, as a means for diagnosing and prognosticating die status of a tissue sample. A wide variety of factors associated with malignancy can be utilized, such as tlie expression of genes associated with malignancy (e.g. PSA, PSCA and PSM expression for prostate cancer etc.) as well as gross cytological observations (see, e.g., Bocking et al., 1984, Anal. Quant. Cytol. 6(2):74-88; Epstein, 1995, Hum. Pathol. 26(2):223-9; Thorson e/ al., 1998, Mod. Pathol. 11(6):543-51 ; Baisden et al., 1999, Am J. Surg. Pathol. 23(8):918-24). Methods for observing a coincidence between tlie expression of 213P1F11 gene and 213P1F11 gene products (or perturbations in 213P1F11 gene and 213P1F11 gene products) and anodier factor that is associated with malignancy are usefiil, for example, because the presence of a set of specific factors that coincide with disease provides information crucial for diagnosing and prognosticating tlie status of a tissue sample.

In one embodiment, methods for observing a coincidence between tl e expression of 213P1F11 gene and 213P1F1 1 gene products (or perturbations in 213P1F11 gene and 213P1F11 gene products) and anodier factor associated with malignancy entails detecting the overexpression of 213P1F1 1 mRNA or protein in a tissue sample, detecting tlie overexpression of PSA mRNA or protein in a tissue sample (or PSCA or PSM expression), and observing a coincidence of 213P1F11 mRNA or protein and PSA mRNA or protein overexpression (or PSCA or PSM expression). In a specific embodiment, the expression of 213P1F11 and PSA niRNA in prostate tissue is examined, where tlie coincidence of 213P1F11 m d PSA mRNA overexpression in tl e sample indicates the existence of prostate cancer, prostate cancer susceptibility or the emergence or status of a prostate tumor.

Methods for detecting and quantifying tlie expression of 213P1F11 mRNA or protein are described herein, and standard nucleic acid and protein detection and quaiitification technologies are well known in tlie art Standard methods for tlie detection and quantification of 213P1F11 mRNA include in situ hybridization using labeled 213P1F11 riboprobes, Northern blot and related techniques using 213 P 1 F 11 polynucleotide probes, RT-PCR analysis using primers specific for 213P1F11, and odier amplification type detection methods, such as, for example, branched DNA, SISB A, TMA and tlie like. In a specific embodiment, semi-quantitative RT-PCR is used to detect and quantify 213P1F11 mRNA expression. Any number of primers capable of amplifying 213P1F11 can be used for this purpose, including but not limited to tl e various primer sets specifically described herein. In a specific embodiment, polyclonal or monoclonal antibodies specifically reactive with the wild-type 213P1F11 protein can be used in an hmnunoliistochemical assay of biopsied tissue.

IX.) Identification of Molecules That Interact With 213P1F11 The 213P1F1 1 protein and nucleic acid sequences disclosed herein allow a skilled artisan to identify proteins, small molecules and other agents that interact with 213PlFl l, as well as pathways activated by 213P1F11 via any one of a variety of art accepted protocols. For example, one can utilize one of tlie so-called interaction trap systems (also referred to as tlie "two-hybrid assay"). In such systems, molecules interact and reconstitute a transcription factor which directs expression of a reporter gene, whereupon tlie expression of tlie reporter gene is assayed. Other systems identify protein-protein interactions in vivo through reconstitution of a eukaryotic transcriptional activator, see, e.g., U.S. Patent Nos. 5,955,280 issued 21 September 1999, 5,925,523 issued 20 July 1999, 5,846,722 issued 8 December 199.8 and 6,004,746 issued 21 December 1999. 42 Algorithms are also available in the art for genome-based predictions of protein function (see, e.g., Marcotte, et ai , Nature 402: 4 November 1999, 83-86).

Alternatively one can screen peptide libraries to identify molecules that interact with 213P1F11 protein sequences. In such metliods, peptides that bind to 213P1F11 are identified by screening libraries tliat encode a random or controlled collection of amino acids. Peptides encoded by the libraries are expressed as fusion proteins of bacteriophage coat proteins, the bacteriophage particles are then screened against the 213P 1F1 1 prolein(s).

Accordingly, peptides having a wide variety of uses, such as therapeutic, prognostic or diagnostic reagents, are thus identified without any prior information on die structure of the expected ligand or receptor molecule. Typical peptide libraries and screening metliods tliat can be used to identify molecules that interact with 213P1F11 protein sequences are disclosed for example in U.S. Patent Nos. 5,723,286 issued 3 March 1998 a d 5,733,731 issued 31 March 1 98.

Alternatively, cell lines tliat express 213P1F11 are used to identify protein-protein interactions mediated by 213P1F1.1. Such interactions can be examined using iiiuiiiuioprecipitation techniques (see, e.g., Hamilton B.J., et at. Biochem. Biophys. Res. Commun. 1999, 261 :646-51). 213P1 F11 protein can be imnuinoprecipitated from 213P1F11-expressing cell lines using anti-213PlFl 1 antibodies. Alternatively, antibodies against His-tag can be used in a cell line engineered to express fusions of 213P1F1 1 and a His-tag (vectors mentioned above). The imnuinoprecipitated complex can be examined for protein association by procedures such as Western blotting, 3iS-methionine labeling of proteins, protein microsequencing, silver staining and two-dimensional gel electrophoresis.

Small molecules and ligands tliat interact with 213P1F11 can be identified through related embodiments of such screening assays. For example, small molecules can be identified tliat interfere with protein function, including molecules that interfere with 213P1F11 's ability to mediate phosphorylation and de-phosphorylation, interaction with DNA or RNA molecules as an indication of regulation of cell cycles, second messenger signaling or lumorigenesis. Similarly, small molecules that modulate 213P lFl l-related ion channel, protein pump, or cell communication functions are identified and used to treat patients tliat have a cancer tliat expresses 213P1F11 (see, e.g., Hille, B., ionic Channels of Excitable Membranes 2nd Ed., Sinauer Assoc., Sunderland, MA, 1992). Moreover, ligands tliat regulate 213P1F1 1 function can be identified based on their ability to bind 213P1F11 and activate a reporter construct. Typical metliods are discussed for example in U.S. Patent No. 5,928,868 issued 27 July 1999, and include metliods for forming hybrid ligands in wl ich at least one ligand is a small molecule. In an illustrative embodiment, cells engineered to express a fusion protein of 213P1F1 1 and a DNA-binding protein are used to co-express a fusion protein of a hybrid ligaiid/small molecule and a cDNA library transcriptional activator protein. The cells further contain a reporter gene, (lie expression of which is conditioned on the proximity of the first and second fusion proteins to each other, an event tliat occurs only if the hybrid ligand binds to target sites on both hybrid proteins. Those cells tliat 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 wliich activate or inhibit 213P 1F1 1.

An embodiment of this invention comprises a method of screening for a molecule tliat interacts with a 213P1F11 amino acid sequence shown in Figure 2 or Figure 3, comprising the steps of contacting a 43 population of molecules with a 213P1F1 1 amino acid sequence, allowing tlie population of molecules and tlie 213P1F11 amino acid sequence to interact under conditions that facilitate an interaction, determining tlie presence of a molecule that interacts with tlie 213P1F11 amino acid sequence, and then separating molecules that do not interact with tlie 213P1F11 amino acid sequence from molecules that do. In a specific embodiment, tlie method further comprises purifying, characterizing and identifying a molecule that interacts with die 213P1F11 amino acid sequence. The identified molecule can be used to modulate a function performed by 213P1F11. In a preferred embodiment, tlie 213P1F11 amino acid sequence is contacted with a library of peptides.

X.) Therapeutic Methods and Compositions The identification of 213P1F11 as a protein that is normally expressed in a restricted set of tissues, but which is also expressed in prostate and other cancers, opens a number of therapeutic approaches to the treatment of such cancers. As contemplated herein, 213P1F11 functions as a transcription factor involved in activating tumor-promoting genes or repressing genes that block tumorigenesis.

Accordingly, therapeutic approaches that inhibit tlie activity of a 213P1F11 protein are useful for patients suffering from a cancer that expresses 213P1F1 1. These therapeutic approaches generally fall into two classes. One class comprises various methods for inhibiting tl e binding or association of a 213P1F1 1 protein with its binding partner or with other proteins. Another class comprises a variety of methods for inhibiting tlie transcription of a 213P1F11 gene or translation of 213P1F11 niRNA.

X.A.) Anti-Cancer Vaccines The invention provides cancer vaccines comprising a 213P1F11 -related protein or 213P1F1 1-related nucleic acid. In view of tlie expression of 213P1F1 1, cancer vaccines prevent and/or treat 213P1F1 1 -expressing cancers with nunimal or no effects on non-target tissues. The use of a tumor antigen in a vaccine that generates humoral and/or 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 al, 1997, J. Immunol. 159:3113-3117).

Such methods can be readily practiced by employing a 213P1 F11-related protein, or a 213P1F11-encoding nucleic acid molecule and recombinant vectors capable of expressing and presenting the 213P1 F11 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 inimunoreactive epitopes are known in the art (see, e.g., Heryln et al. , Ann Med 1999 Feb 31(l):66-78; Maruyama et al., Cancer Immunol Iminunother 2000 Jun 49(3): 123-32) Briefly, such methods of generating an immune response (e.g. humoral and/or cell-mediated) in a mammal, comprise tlie steps of: exposing tlie mammal's immune system to an inimunoreactive epitope (e.g. an epitope present in a 213P1F11 protein shown in Figure 3 or analog or homolog thereof) so that tlie mammal generates ait immune response that is specific for that epitope (e.g. generates antibodies that specifically recognize dial epitope). In a preferred method, a 213P1F1 1 immunogen contains a biological motif, see e.g., Tables V-XIX, or a peptide of a size range from 213P1F1 1 indicated in Figure 5, Figure 6, Figure 7, Figure 8, and Figure 9.

The entire 213P1F11 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, 44 A. et al. , J. Clin. Invest. 95:341, 1995), peptide compositions encapsulated in poly(DL-lactide-co-glycolide) ("PLG") microspheres (see, e.g. , Eldridge, et al. , Mokc. Immunol. 28:287-294, 1991 : Alonso et al , Vaccine 12:299-306, 1994; ones et al. , Vaccine 13:675-681, 1995), peptide compositions contained in immune stimulating complexes (1SCOMS) (see, e.g., Takahashi et al., Nature 344:873-875, 1990; Hu et al., Clin Exp Immunol. 113 :235-243, 1998), multiple antigen peptide systems (MAPs) (see e.g. , Tarn, J. P., Proc. Natl. Acad. Sci. U.S.A. 85:5409-5413, 1988; Tarn, J.P., J. Immunol. Methods 196: 17-32, 1996), peptides formulated as multivalent peptides; peptides for use in ballistic delivery systems, typically crystallized peptides, viral delivery vectors (Perkus, M. E. et al , In: Concepts in vaccine development, Kaufmann, S. H. E., ed., p. 379, 1996; Chakrabarti, S. et al , Nature 320:535, 1986; Hu, S. L. et al, Nature 320:537, 1986; Kieny, M.-P. et al , AIDS Bio/Technology 4:790, 1986; Top, F. H. et al , J. Infect. Dis. 124: 148, 1971 ; Chanda, P. K. et al, Virology 175:535, 1990), particles of viral or synthetic origin (e.g. , Kofler, N. et al, J. Immunol Methods. 192 :25, 1996; Eldridge, J. H. et al. , Sem. Hematol. 30: 16, 1993; Falo, L. D., Jr. et al , Nature Med. 7:649, 1995), adjuvants (Warren, H. S., Vogel, F. R., and Chedid, L. A. Aivni. 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. L., Immunol Today 17: 131, 1996), or, naked or particle absorbed cDNA (Ulnier, J. B. et al , Science 259: 1745, 1993; Robinson, H. L., Hunt, L. A., and Webster, R. G., Vaccine 11 :957, 1993; Shiver, 5. W. et al, In: Concepts in vaccine development, Kaufmann, S. H. E., ed., p. 423, 1996; Cease, K. B., and Berzofsky, J. A., Anriu. Rev. Immunol. 12:923, 1994 and Eldridge, J. H. et al, Sent. Hematol. 30: 16, 1993). Toxin-targeted delivery technologies, also known as receptor mediated targeting, such as those of Avant Immunotherapeutics, Inc. (Needham, Massacluisetls) may also be used.

In patients with 213P1F11-associated cancer, t e vaccine compositions of the invention can also be used in conjunction with other treatments used for cancer, e.g., 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 213P1F11 protein Uial bind corresponding HLA alleles (see e.g., Table IV; Epimer™ and Epimatrix™, Brown University (located at the World Wide Web .brovvn.edii/ResearclvTB-HlV_Lab/epiniau-ix/epimatrix.htinl); mid, BIMAS, (URL bimas.dcrt.mh.gov/; SYFPEITHI at URL syfpeithi.bnii-heidelberg.com/). In a preferred embodiment, a 213P1F11 immunogen contains one or more amino acid sequences identified using techniques well known in the art, such as the sequences shown in Tables V-X1X, or a peptide of 8, 9, 10 or 11 amino acids specified by an HLA Class I motif/supermotif (e.g., Table IV (A), Table IV (D), or Table IV (E)) and/or a peptide of at least 9 amino acids that comprises an HLA Class II motif/supermotif (e.g., Table IV (B) or Table IV (C)). 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 11 molecule. Due to (l e binding groove differences between HLA Class I and II, HLA Class 1 motifs are length specific, 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 acid positions in a Class II motif are relative only to each oilier, not the overall peptide, i.e., additional amino acids, can be 45 attached to the amino and/or carboxyl termini of a motif-bearing sequence. ' HLA Class II epitopes are often 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids long, or longer than 25 amino acids.

Autibody-based Vaccines 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 tire mammal's immune system to an immunogenic epitope on a protein (e.g. a 213P1F1 1 protein) so that an immune response is generated. A typical embodiment consists of a method for generating an immune response to 213P1F11 in a host, by contacting the host with a sufficient amount of at least one 213P1F11 B cell or cytotoxic T-cell epitope or analog thereof; and at least one periodic interval .thereafter re-contacting the host with the 213P1F11 B cell or cytotoxic T-cell epitope or analog thereof. A specific embodiment consists of a method of generating an immune response against a 213PlFl l-related protein or a man-made multiepitopic peptide comprising: administering 213P1F11 immunogen (e.g. a 213P1F11 protein or a peptide fragment thereof, a 213P1F11 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, e.g., U.S. Patent No. 6, 146,635) or a universal helper epitope such as a PADRE™ peptide (Epimmune Inc., San Diego, CA; see, e.g., Alexander et al. . Immunol. 2000 164(3); 164(3): 1625-1633; Alexander e/ o/. , Immunity 1994 1(9): 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 213P 1 F11 immunogen by: administering in vivo to muscle or skin of the individual's body a DNA molecule that comprises a DNA sequence t at encodes a 213P1F11 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, e.g., 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 antiidiolypic antibody can be administered thai nunucs 213P1F1 1, 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 213PIF11. Constructs comprising DNA encoding a 213P 1F11 -related protein/immunogen and appropriate regulatory sequences can be injected directly into muscle or skin of an individual, such thai the cells of the muscle or skin take-up the construct and express the encoded 213P1F1 1 protein/immunogen. Alternatively, a vaccine comprises a 213P1F11-related protein. Expression of the 213P lFl l-related protein immunogen results in the generation of prophylactic or therapeutic humoral and cellular immunity against cells mat bear a 213P 1F11 protein. Various prophylactic and therapeutic genetic immunization techniques known in tire art can be used (for.review, see iiuonnation and references published at Internet address located at live World Wide Web .genweb.com). Nucleic acid-based delivery is described, for instance, in Wolff et. at. , Science 247: 1465 (1990) as well as U.S. Patent Nos. 5,580,859; 5,589,466; 46 ,804,566; 5,739, 118; 5,736,524; 5,679,647; WO 98/04720. Examples of DNA-based delivery technologies include "naked DNA", facilitated (bupivicaine, polymers, peptide-mediated) deliver}', cationic lipid complexes, and particle-mediated ("gene gnu") or pressure-mediated delivery (see, e.g. , 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-associated vims, lentivirus, and sindbis virus (see, e.g., Restifo, 1996, Curr. Opiii. Immunol. 8:658-663; Tsang et at. J. Natl. Cancer Inst. 87:982-990 (1995)). Non-viral delivery systems can also be employed by introducing naked DNA encoding a 213PlFl l-related protein into the patient (e.g., intramuscularly or intradermally) to induce an anti-tumor response.

Vaccinia virus is used, for example, as a vector to express nucleotide sequences that encode the peptides of the invention. Upon introduction into a host, the recombinant vaccinia virus expresses the protein immunogenic peptide, and thereby elicits a host immune response. Vaccinia vectors and methods useful in immunization protocols are described in, e.g., U.S. Patent No. 4,722,848. Anodier 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 mid adeno-associated 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.

Thus, gene delivery systems are used to deliver a 213P1F11 -related nucleic acid molecule. In one embodiment, the full-length human 213P1F1 1 cDNA is employed, hi another embodiment, 213P1F11 nucleic acid molecules encoding specific cytotoxic T lymphocyte (CTL) and/or antibody epitopes are employed.

Ex Vivo Vaccines Various ex vivo strategies can also be employed to generate an immune response. One approach involves the use of antigen presenting cells (APCs) such as dendritic cells (DC) to present 213PIF11 antigen to a patient's immune system. Dendritic cells express MHC class I and Π molecules, B7 co-stimulator, and 1L-12, mid 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 213P1F11 peptides to T cells in the context of MHC class I or II molecules. In one embodiment, autologous dendritic cells are pulsed with 213P1F1 1 peptides capable of binding to MHC class I and/or class II molecules. In another embodiment, dendritic cells are pulsed with (lie complete 213P1F11 protein. Yet another embodiment involves engineering the overexpression of a 213P1F11 gene in dendritic cells using various implementing vectors known in the art, such as adenovirus (Arthur e/ al , 1997, Cancer Gene Ther. 4: 17-25), retrovirus (Henderson et al., 1996, Cancer Res. 56:3763-3770), lentivirus, adeno-associated vims, DNA transfection (Ribas et al. , ' 1997, Cancer Res. 57:2865-2869), or tumor-derived RNA transfection (Ashley et al , 1997, J. Exp. Med. 186: 1177-1 182). Cells diat express 213P 1F11 can also be engineered to express immune modulators, such as GM-CSF, mid used as immunizing agents.

X.B.) 213P1F11 as a Target for Antibody-based Therapy 47 213P1F1 1 is an attractive target for antibody-based therapeutic strategies. A number of antibody strategies axe known in the art for targeting both extracellular and intracellular molecules (see, e.g., complement and ADCC mediated killing as well as the use of intrabodies). Because 213P1F1 1 is expressed by cancer cells of various lineages relative to corresponding normal cells, systemic administration of 213PlFl l-imnumoreactive compositions are prepared that exhibit excellent sensitivity without toxic, nonspecific and/or non-target effects caused by binding of the immunoreactive composition to non-target organs and tissues. Antibodies specifically reactive with domains of 213P1F11 are useful to treat 213P1F11-expressing cancers systemic-ally, either as conjugates with a toxin or therapeutic agent, or as naked antibodies capable of inliibiting cell proliferation or function. 213P1F1 1 antibodies can be introduced into a patient such that the antibody binds to 213P1F11 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 213P1F1 1 , inliibition of ligand binding or signal transduction pathways, modulation of tumor cell differentiation, alteration of tumor angiogenesis factor profiles, and/or apoptosis.

Those skilled in the art understand that antibodies can be used to specifically target and bind immunogenic molecules such as an immunogenic region of a 213P1F11 sequence shown in Figure 2 or Figure 3. In addition, skilled artisans understand that it is routine to conjugate antibodies to cytotoxic agents (see, e.g., 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 to antibodies specific for a molecule expressed by (hat cell (e.g. 213P1F11), the cytotoxic agent will exert its known biological effect (i.e. cytotoxicity) on those' cells.

A wide variety of compositions and methods for using antibody-cytotoxic agent conjugates to kill cells are known in tire art. In the context of cancers, typical methods entail administering to an animal having a tumor a biologically effective amount of a conjugate comprising a selected cytotoxic and/or therapeutic agent linked to a targeting agent (e.g. an anti-213PlFl I antibody) that binds to a marker (e.g. 213P1F1 1) ■ expressed, accessible to binding or localized on the cell surfaces. A typical embodiment is a method of delivering a cytotoxic and/or therapeutic agent to a cell expressing 213P1F11, comprising conjugating the cytotoxic agent to an antibody that immunospecifically binds to a 213P1F11 epitope, and, exposing tire 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-213PlFH 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 et al. , 1997, Blood 90:3179-3 186, Tsunenari et al. , 1997, Blood 90:2437-2444), gastric cancer (Kasprzyk et al , 1992, Cancer Res. 52:2771-2776), B-cell lymphoma (Funakoshi et al. , 1996, J. Immunother. Emphasis Tumor Immunol. 19:93- 101), leukemia (Zhon ei a/. , :1996, Leuk. Res. 20:581 -589), colorectal cancer (Moun 48 et ai, 1994, Cancer Res. 54:6160-6166; Velders et al, 1995, Cancer Res. 55:4398-4403), and breast cancer (Shepard et al , 1991 , J. Clin. Iininunol. 11 : 1 17-127). Some tlierapeutic approaches involve conjugation of naked antibody to a toxin or radioisotope, such as the conjugation of Y91 or I131 to anti-CD20 antibodies (e.g., Zevalin™, IDEC Pharmaceuticals Corp. or Bexxar™, Coulter Pharmaceuticals), while otiiers involve coadministration of antibodies and other tlierapeutic agents, such as Herceptin™ (trastuzuniab) with paclitaxel (Genentech, Inc.). The antibodies can be conjugated to a tlierapeutic agent. To treat prostate cancer, for example, 213P1F11 antibodies can be administered in conjunction with radiation, chemotherapy or hormone ablation. Also, antibodies can be conjugated to a toxin such as calicheamicin (e.g., Mylotarg™, Wyeth-Ayerst, Madison, NJ, a recombinant humanized IgG4 kappa antibody conjugated to antitumor antibiotic calicheamicin) or a niaytaiisinoid (e.g., taxane-based Tumor-Activated Prodrug, TAP, platform, ImniunoGen, Cambridge, MA, also see e.g., US Patent 5,416,064).

Aldiough 213P1F11 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 cliemotherapeutic or radiation regimen for patients who have not received cliemotherapeutic 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 cliemotherapeutic agent very well. Fan et al. (Cancer Res. 53 :4637-4642, 1993), Prewett et al. (International J. of Onco. 9:217-224, 1996), and Hancock et al. (Cancer Res. 51 :4575-4580, 1991) describe die use of various antibodies together widi cliemotherapeutic agents.

Although 213P1F11 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 cliemotherapeutic or radiation regimen for patients who have not received cliemotherapeutic treatment. Additionally, antibody therapy can enable Uie use of reduced dosages of concomitant chemotherapy, particularly for patients who do not tolerate the toxicity of the chemodierapeutic agent very well.

Cancer patients can be evaluated for die presence and level of 213P1F1 1 expression, preferably using immiuiohistochemical assessments of tumor tissue, quantitative 213P1F11 imaging, or odier techniques that reliably indicate the presence and degree of 213P 1 F11 expression. Inmiunohistoclieniical analysis of tumor biopsies or surgical specimens is-preferred for this purpose. Methods for immunoliistocheinical analysis of tumor tissues are well known in die art.

Anti-213P1F11 monoclonal antibodies that treat prostate and other cancers include those that initiate a potent immune response against the tumor or those dial are directly cytotoxic. In tiiis regard, anti-213PlFl 1 monoclonal antibodies (mAbs) can elicit tumor cell lysis by either complement-mediated or antibody-dependent cell cytotoxicity (ADCC) mechanisms, both of which require an intact Fc portion of the immunoglobulin molecule for interaction with effector cell Fc receptor sites on complement proteins. In addition, anti-213PlFl l mAbs tliat exert a direct biological effect on tumor growth are useful to treat cancers tiiat express 213P1F11. Meclianisnis by which directly cytotoxic mAbs act include: inl ibition of cell growth, modulation of cellular differentiation, modulation of tumor angiogenesis factor profiles, and the -induction of 49 apoptosis. The mechanism(s) by which a particular anti-213P1F11 niAb exerts an anti-tumor effect is evaluated using any number of in vitro 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 monoclon l antibodies, or human/mouse chimeric mAbs can induce moderate to strong immune responses against tire 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 tlierapeutic methods of the invention are those that are either fully human or humanized and d at bind specifically to the target 213P1F11 antigen with high affinity but exhibit low or no antigenicity in the patient.

Therapeutic methods of the invention contemplate the administration of single anti-213P1 F11 mAbs as well as combinations, or cocktails, of different mAbs. Such mAb cocktails can have certain advantages inasmuch as they contain mAbs diat 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 tlierapeutic effects. In addition, anti-213P1F11 mAbs can be administered concomitantly with other tlierapeutic modalities, including but not limited to various chemodierapeutic agents, androgen-blockers, immune modulators (e.g., IL-2, GM-CSF), surgery or radiation. The anti-213P1F11 mAbs are administered in their "naked" or unconjugated form, or can have a tlierapeutic agent(s) conjugated to them.

Anti-213P1F11 antibody formulations are administered via any route capable of delivering die antibodies to a tumor cell. Routes of administration include, but are not limited to, intravenous, intraperitoneal, intramuscular, intratumor, intradermal, and die like. Treatment generally involves repeated administration of the anti-213PlFl l antibody preparation, via an acceptable route of administration such as intravenous injection (IV), typically at a dose in the range of about 0.1, .2, .3, .4, .5, .6, .7, .8, .9., 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 nig mAb per week are effective and well tolerated.

Based on clinical experience with the Herceptin™ 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-213P1F11 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 die 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, Hie binding affinity and half life of the Ab or mAbs used, the degree of 213P1F1 1 expression in the patient, the extent of circulating shed 213P1F11 antigen, the desired steady-state antibody concentration level, frequency of treatment, and die influence of chemodierapeutic or odier agents used in combination wiUi die treatment method of the invention, as well as the health status of a particular patient.

Optionally, patients should be evaluated for die levels of 213P1F11 in a given sample (e.g. die levels of circulating 213P1F11 antigen and/or 213P1F11 expressing cells) in order to assist in the determination of die most effective dosing regimen, etc. Such evaluations are also used for monitoring purposes throughout 50 •therapy, and are useful to gauge therapeutic success in combination with the evaluation of other parameters (for example, urine cytology and/or ImmunoCyt levels in bladder cancer therapy, or by analogy, serum PSA levels in prostate cancer therapy).

Anti-idiotypic anti-213PlFl 1 antibodies can also be used in anti-cancer therapy as a vaccine for inducing an immune response to cells expressing a 213PlFl l-related protein. In particular, tlie generation of anti -idiotypic antibodies is well known in tlie art; this methodology can readily be adapted to generate anti-idiotypic anti-213PlFl 1 antibodies that mimic an epitope on a 213P1F11-related protein (see, for example, Wagner et al, 1997, Hybridoma 16: 33-40; Foon et al, 1995, J. Clin, invest. 96:334-342; Herlyn et at., 1996, Cancer Immunol. Immunother. 43 :65-76). Such an anti-idiotypic antibody can be used in cancer vaccine strategies.

X.C.) 213P1F11 as a Target for Cellular Immune Responses Vaccines and methods of preparing vaccines that contain an immunogenically effective amount of one or more HL A-biiiding peptides as described herein are further embodiments of tlie invention.

Furthermore, vaccines in accordance with tl e invention encompass compositions of one or more of t e claimed peptides. A peptide can be present in a vaccine individually. Alternatively, tl e peptide can exist as a omopolymer comprising multiple copies of the same peptide, or as a heteropolymer of various peptides. Polymers have tlie advantage of increased immunological reaction and, where different peptide epitopes are used to make up tlie polymer, tlie additional ability to induce antibodies and/or CTLs that react with different antigenic determinants of tlie 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, e.g. , recombinantly or by chemical synthesis.

Carriers that can be used with vaccines of the invention are well known in the art, and include, e.g., tliyroglobuliii, albumins such as human senun albumin, tetanus toxoid, polyamino acids such as poly L-lysine, poly L-glutamic acid, influenza, hepatitis B virus core protein, and (lie like. The vaccines can contain a physiologically tolerable (i.e., 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 tlie art.

Additionally, as disclosed herein, CTL responses can be primed by conjugating peptides of the invention to lipids, such as tripalnutoyl-S-glycerylcysteinlyseryl- serine (P3CSS). Moreover, an adjuvant such as a synthetic cytosine-phosphorotluolated-guanine-contaimng (CpG) oligonucleotides has been found to increase CTL responses 10- to 100-fold, (see, e.g. Davila and Celis, J. Immunol. 165:539-547 (2000)) Upon immunization with a peptide composition in accordance with tlie invention, via injection, aerosol, oral, transdermal, transimicosal, intrapleural, intrathecal, or other suitable routes, tlie immune system of tlie host responds to tl e vaccine by producing large amounts of CTLs and/or HTLs specific for tlie desired antigen. Consequently, tlie host becomes at least partially immune to later development of cells tiiat express or overexpress 213P1F11 antigen, or derives at least some therapeutic benefit when the antigen was tumor-associated.

In some embodiments, it may be desirable to combine tlie class I peptide components with components that induce or facilitate neutralizing antibody and or helper T cell responses directed to tlie target antigen. A preferred embodiment of such a composition comprises class I mid class II epitopes in accordance 51 with the invention. An alternative embodiment of such a composi tion comprises a class 1 and/or class II epitope in accordance with the invention, along with a cross reactive HTL epitope such as PADRE™ (Epimmune, San Diego, CA) molecule (described e.g., in U.S. Patent Number 5,736,142).

A vaccine of the invention can also include antigen-presenting cells (APC), such as dendritic cells (DC), as a vehicle to present peptides of tire 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, e.g., with a minigene in accordance with the invention, or are pulsed with peptides. The dendritic cell can then be administered, 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 tire 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 tire native antigen from which the epitopes are derived. 1. ) Epitopes are selected which, upon administration, mimic immune responses that have been obsen'ed to be correlated with tumor clearance. 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, e.g. , Rosenberg et al., 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. 2. ) Epitopes are selected that have the requisite binding affinity established to be correlated with immunogenicity: for HLA Class I an IC50 of 500 nM or less, often 200 nM or less; and for Class II an IC50 of 1000 nM or less. 3. ) Sufficient supermotif bearing-peplides, 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. 4. ) 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 mulli-epitopic sequence, such as a sequence comprising nested epitopes, it is generally important to screen tire sequence in order to insure that it does not have pathological or other deleterious biological properties. 52 6. ) If a polyepitopic protein is created, or when creating a minigene, an objective is to generate tlie smallest peptide that encompasses tlie epitopes of interest. Tins principle is similar, if not tlie same as that employed when selecting a peptide comprising nested epitopes. However, with an artificial polyepitopic peptide, the size minimization objective is balanced against tlie need to integrate any spacer sequences between epitopes in tlie polyepitopic protein. Spacer amino acid residues can, for example, be introduced to avoid junctional epitopes (an epitope recognized by the immune system, not present in tlie target antigen, and only created by tlie man-made juxtaposition of epitopes), or to facilitate cleavage between epitopes and thereby enhance epitope presentation, junctional epitopes are generally to be avoided because the recipient may generate mi immune response to that non-native epitope. Of particular concern is a junctional epitope tliat is a "dominant epitope." A dominant epitope may lead to such a zealous response that immune responses to other. epitopes are diminished or suppressed. 7. ) Where tlie sequences of multiple variants of the same target protein are present, potential peptide epitopes can also be selected on (lie basis of their conservancy. For example, a criterion for conservancy may define that tlie entire sequence of an HLA class 1 binding peptide or tlie entire 9-mer core of a class II binding peptide be conserved in a designated percentage of tlie sequences evaluated for a specific protein antigen.

X.C.1. Minigene Vaccines A number of different approaches are available which allow simultaneous delivery of multiple epitopes. Nucleic acids encoding tl e peptides of tlie invention are a particularly useful embodiment of tlie invention. Epitopes for inclusion in a minigene are preferably selected according to tlie guidelines set forth in tlie previous section. A preferred means of administering nucleic acids encoding tlie peptides of tlie invention uses minigene constructs encoding a peptide comprising one or multiple epitopes of tlie invention.

The use of multi-epitope minigenes is described below and in, lshioka et al., J. Immunol. 162:3915-3925, 1999; An, L. and Whitton, J. L., J. Virol. 71 :2292, 1997; Thomson, S. A. et al., J. Immunol. 157:822, 1996; Whitton, J. L. et al. , J. Virol. 67:348, 1993; Hanke, R. et al. , Vaccine 16:426, 1998. For example, a multi-epitope DNA plasniid encoding supermotif- and/or motif-bearing epitopes derived 213P1F1 1, tlie PADRE© universal helper T cell epitope or multiple HTL epitopes from 213P1F1 1 (see e.g., Tables V-XIX), and an endoplasmic reticuluni-translocating signal sequence can be. engineered. A vaccine may also comprise epitopes that are derived from other TAAs.

The imiiumogeiiicity of a inultirepitopic minigene can be confirmed in transgenic mice to evaluate tlie magnitude of CTL induction responses against (lie epitopes tested. Further, tlie immunogenicity of DNA-encoded epitopes in vivo can be correlated with tlie in vitro responses of specific CTL lines against target cells transfected with tire DNA plasniid. Thus, these experiments can show that the minigene serves to both: 1.) generate a CTL response and 2.) that tlie induced CTLs recognized cells expressing tlie encoded epitopes.

For example, to create a DNA sequence encoding tlie selected epitopes (minigene) for expression in human cells, tlie amino acid sequences of the epitopes may be reverse translated. A human codon usage table can be used to guide tlie codon choice for each amino acid. These epitope-encoding DNA sequences may be directly adjoined, so diat 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 annuo acid sequences that can be reverse translated and included in tlie minigene sequence 53 include: HLA class I epitopes ,'HL A class 11 epitopes, antibody epitopes, a ubiquitination signal sequence, and/or an endoplasmic reticulum targeting signal. In addition, HLA presentation of CTL and HTL epitopes may be improved by including synthetic (e.g. poly-alanine) or naturally-occurring flanking sequences adjacent to tire CTL or HTL epitopes; these larger peptides comprising tlie epitope(s) are within tire, scope of tlie invention.

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

Standard regulatory sequences well known to those of skill in tl e art are preferably included in tl e vector to ensure expression in tlie target cells. Several vector elements are desirable: a promoter with a downstream cloning site for minigene insertion; a polyadenylation signal for efficient transcription termination; an E. coli origin of replication; and an E. coli selectable marker (e.g. ampicillin or kanamycin resistance).

Numerous promoters can be used for this purpose, e.g., die human cytomegalovirus (hCMV) promoter. See, e.g. , U.S. Patent Nos. 5,580,859 and 5,589,466 for oilier suitable promoter sequences.

Additional vector modifications may be desired to optimize minigene expression and inimunogeiiicity. In some cases, introns are required for efficient gene expression, and one or more synthetic or naturally-occurring introns could be incorporated into tlie transcribed region of the minigene. The inclusion of niRNA stabilization sequences and sequences for replicatioii in mammalian cells may also be considered for increasing minigene expression.

Once an expression vector is selected, tlie minigene is cloned into tlie polylinker region downstream of the promoter. Tliis plasinid is transformed into an appropriate E. coli strain, and DNA is prepared using standard techniques. The orientation and DNA sequence of the minigene, as well as all other elements included in tlie vector, are confirmed using restriction mapping and DNA sequence analysis. Bacterial cells harboring tlie correct plasinid can be stored as a master cell bank and a working cell bank.

In addition, immuiiostimulatory sequences (ISSs or CpGs) appear to play a role in tlie iinmunogenicity of DNA vaccines. These sequences may be included in tlie vector, outside tlie minigene coding sequence, if desired to enhance imniunogenicity.

In some embodiments, a bi-cislronic expression vector which allows production of both tlie minigene-encoded epitopes and a second protein (included to enhance or decrease iinmunogenicity) can be used. Examples of proteins or polypeptides that could beneficially enhance tlie immune response if co-expressed include cytokines (e.g., IL-2, IL-12, GM-CSF), cylokine-induciiig molecules (e.g., LeIF), costimulatory molecules, or for HTL responses, pan-DR binding proteins (PADRE™, Epin miuie, San Diego, CA). Helper (HTL) epitopes can be joined to intracellular targeting signals and expressed separately from expressed CTL epitopes; tliis allows direction of tlie HTL epitopes to a cell compartment different than that of tlie CTL epitopes. If required, tliis could facilitate more efficient entry of HTL epitopes into tl e HLA class II patliway, tliereby improving HTL induction. In contrast to HTL or CTL induction, specifically decreasing tlie immune response by co-expression of immunosuppressive molecules (e.g. TGF-β) may be beneficial in certain diseases. 54 Therapeutic quantities of plasniid DNA can be produced for example, by fermentation in E coli, followed by purification. Aliqnots from the working cell bank are used to inoculate growth medium, and grown to saturation in shaker flasks or a bioreacior according to well-known techniques. Plasniid DNA can be purified using standard bioseparation 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 reconslitution.of lyophilized DNA in sterile phosphate-buffer saline (PBS). Tliis approach, known as "naked DNA," is currently being used for intramuscular (1M) administration in clinical trials. To maximize the imniunotherapeutic effects of minigene DNA vaccines, an alternative 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, e.g., as described by WO 93/24640; Mannino & Gould-Fogerite, BioTechniques 6(7): 682 (1988); U.S. Pat No. 5,279,833; WO 91/06309; and Feigner, et ai, Proc. Nat'lAcad. Sci. USA 84:7413 (1987). In addition, peptides and compounds referred to collectively as protective, interactive, non-condensing compounds (PINC) could also be complexed to purified plasmid DNA to influence variables such as stability, intramuscular dispersion, or trafficking to specific organs or cell types.

Target cell sensitization can be used as a functional assay for expression and HLA class I presentation of minigene-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. Eleclroporation can be used for "naked" DNA, whereas cationic lipids allow direct in vitro transfection. A plasmid expressing green fluorescent protein (GFP) can be co-transfecled to allow enrichment of transfected cells using fluorescence activated cell sorting (FACS). These cells are then chromium-51 (51 Cr) labeled and used as target cells for epitope-specific CTL lines; cytolysis, detected by 51 Cr release, indicates both production of, and HLA presentation of, minigene-encoded CTL epitopes. Expression of HTL epitopes may be evaluated in an analogous maimer using assays to assess HTL activity.

In vivo immunogeiiicity 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 (e.g., 1M for DNA in PBS, intraperitoneal (i.p.) 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 'Cr-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. ImmunogeniciLy of HTL epitopes is confirmed in transgenic mice in an analogous maimer.

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 particles, such as gold particles. 55 Minigenes can also be delivered using oilier bacterial or viral delivery systems well known in tl e art, e.g. , an expression construct encoding epitopes of tl e invention can be incorporated into a viral vector such as vaccinia.

X.C.2. Combinations of CTL Peptides with Helper Peptides Vaccine compositions comprising CTL peptides of tlie invention can be modified, e.g., analoged, to provide desired attributes, such as improved serum half life, broadened population coverage or enhanced immui ogenicity.

For instance, tlie ability of a peptide to induce CTL activity can be enhanced by linking die 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, e.g., Ala, Gly, or other neutral spacers of nonpolar amino acids or neutral polar amino acids, it will be understood that tlie optionally present spacer need not be comprised of the same residues and thus may be a hetero- or homo-oligomer. When present, tlie spacer will usually be at least one or two residues, more usually tlvree to six residues and sometimes 10 or more residues. The CTL peptide epitope can be linked to tlie T helper peptide epitope either directly or via a spacer either at d e amino or carboxy terminus of the CTL peptide. The amino terminus of either tlie immunogenic peptide or die T helper peptide may be acylated.

In certain embodiments, d e T helper peptide is one that is recognized by T helper cells present in a majority of a genetically diverse population. This can be accomplished by selecting peptides diat bind to many, most, or all of the HLA class II molecules. Examples of such amino acid bind many HLA Class II molecules include sequences from antigens such as tetanus toxoid at positions 830-843 (QYIKANSKFIGITE; SEQ ID NO: 38), Plasmodium falciparum circumsporozoite (CS) protein at positions 378-398 (DIEKKIAKMEKASSVFN WNS; SEQ ID NO: 39), and Streptococcus 18kD protein at positions 116-131 (GAVDSILGGVATYGAA; SEQ ID NO: 40). Other examples include peptides bearing a DR 1 -4-7 superniotif, or either of tlie DR3 motifs.

Alternatively, 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, e.g., PCT publication WO 95/07707). These synthetic compounds called Pan-DR-binding epitopes (e.g., PADRE™, Epimmune, Inc., San Diego, CA) are designed to most preferably bind most HLA-DR (human HLA class II) molecule!. For instance, a pai -DR-binding epitope peptide having die formula: aKXVAAWTLKAAa (SEQ ID NO: 41), where "X" is either cycloliexylalaiiine, phenylalanine, or tyrosine, and a is either D-alanine or L-alanine, has been found to bind to most HLA-DR alleles, and to stimulate die response of T helper lymphocytes from most individuals, regardless of their HLA type. An alternative of a pan-DR binding epitope comprises all "L" natural amino acids and can be provided in the form of nucleic acids that encode the epitope.

HTL peptide epitopes ca also be modified to alter their biological properties. For example, they can be modified to include D-amino acids to increase their resistance to proteases and thus extend their serum half life, or diey can be conjugated to other molecules such as lipids, proteins, carbohydrates, and tlie like to 56 increase their biological activity. For example, a T helper peptide can be conjugated to one or more palmitic acid 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. Lipids have been identified as agents capable of priming CTL in vivo. For example, palmitic acid residues can be attached to the ε-and a-amino groups of a lysine residue and then linked, e.g., via one or more linking residues such as Gly, Gly-Gly-, Ser, Ser-Ser, or the like, to an immunogenic peptide. The lipidated peptide can then be administered either directly in a micelle or particle, incorporated into a liposome, or emulsified in ait adjuvant, e.g., incomplete Freund's adjuvant. In a preferred embodiment, a particularly effective immunogenic composition comprises palmitic acid attached to e- and a- amino groups of Lys, which is attached via linkage, e.g., Ser-Ser, to the amino terminus of the immunogenic peptide.

As another example of lipid priming of CTL responses, E. coli lipoproteins, such as iripalmitoyl-S-glycerylcysteinlyseryl- serine (P3CSS) can be used to prime virus specific CTL when covalently attached to an appropriate peptide {see, e.g., Deres, et al, Nature 342:561, 1989). Peptides of the invention can be coupled to P3CSS, for example, and the lipopeptide administered to an individual to specifically prime an immune response to the target antigen. Moreover, because the induction of neutralizing antibodies can also be primed witli P3CSS-conjugated epitopes, two such compositions can be combined to more effectively elicit both humoral and cell-mediated responses.

X.C.4. Vaccine Compositions Comprising DC Pulsed with CTL nnuVur 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™ (Pharmacia-Monsanto, St. Louis, MO) or GM-CSF/TL-4. After pulsing the DC witli peptides and prior to reinfusion into patients, the DC are washed to remove unbound peptides. In this embodiment, a vaccine comprises peptide-pulsed DCs which present d e pulsed peptide epitopes complexed with HLA molecules on their surfaces.

The DC can be pulsed ex vivo witli a cocktail of peptides, some of which stimulate CTL responses to 213P1F1 1. Optionally, a helper T cell (HTL) peptide, such as a natural or artificial loosely restricted HLA Class II peptide, can be included to facilitate the CTL response. Thus, a vaccine in accordance witli the invention is used to treat a cancer which expresses or overexpresses 213P1F11.

X.D. Adoptive Immunotherapy Antigenic 213PlFl l-related peptides are tised 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 not respond to a therapeutic vaccine peptide or nucleic acid in accordance with the invention. Ex vivo CTL or HTL responses to a particular antigen are induced by incubating in tissue culture the patient's, or genetically compatible, CTL or HTL precursor cells together witli 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 activated 57 and expanded into effector cells, tlie cells are infused back into tlie patient, where tlie}' will destroy (CTL) or facilitate destruction (HTL) of their specific target cell (e.g., a tumor cell). Transfected dendritic cells may also be used as 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 213P1F1 1. In therapeutic applications, peptide and/or nucleic acid compositions are administered to a patient in an amount sufficient to elicit an effective B cell, CTL and/or HTL response to tlie antigen and to cure 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, e.g., tlie particular composition administered, tlie maimer of administration, tlie stage and severity of tlie disease being treated, tlie weight and general state of health of the patient, and tlie judgment of tlie prescribing physician.

For pharmaceutical compositions, tlie immunogenic peptides of the invention, or DNA encoding them, are generally administered to an individual already bearing a tumor that expresses 213P1F11. The peptides or DNA encoding them can be administered individually or as fusions of one or more peptide sequences. Patients can be treated with tlie immunogenic peptides separately or in conjunction widi other treatments, such as surgery, as appropriate.

For therapeutic use, administration should generally begin at tlie first diagnosis of 213P1F11-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 (i. e., including, but not limited to embodiments such as peptide cocktails, polyepitopic polypeptides, minigenes, or TAA-specific CTLs or pulsed dendritic cells) delivered to tile patient may vary according to tlie stage of the disease or the patient's health status. For example, in a patient with a tumor that expresses 213P1F11, a vaccine comprising 213P1 F11 -specific CTL may be more efficacious in killing tumor cells in patient with advanced disease than alternative embodiments.

It is generally important to provide an amount of the peptide epitope delivered by a mode of administration sufficient to effectively stimulate 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 tlie lower value is about 1, 5, 50, 500, or 1,000 pg and tlie 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 1.0 ^tg to about 50,000 pg of peptide pursuant to a boosting regimen over weeks to months may be administered depending upon tlie patient's response and condition as determined by measuring the specific activity of CTL and HTL obtained from tlie 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 methodologies known in tlie art.

In certain embodiments, tlie peptides and compositions of the present invention are employed in serious disease states, that is, life-tlirealening or potentially life threatening situations. In such cases, as a result of tl e minimal amounts of extraneous substances and tlie relative nontoxic nature of tlie peptides in 58 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 relati ve 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, or 1000 yvg and the higher value is about 10,000; 20,000; 30,000; or 50,000 ng. -Dosage values for a human typically range from about 500 μg to about 50,000 yg per 70 kilogram patient. This is followed by boosting dosages of between about 1.0 ^i to about'50,000 ng 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 HTL 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 {e.g. as a cream or topical ointment) administration. Preferably, the pharmaceutical compositions are administered parentally, e.g., 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 aqueous carrier.

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

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

The concentration, of peptides of the invention in the pharmaceutical formulations can vary widely, i.e., 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 carrier, in one embodiment an aqueous carrier, and is administered in a volume/quantity that is known by those of skill in the art to be used for administration of such compositions to humans {see, e.g. , Remington's Pharmaceutical Sciences, 17Ul 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 for a 70 kg patient. For example, for nucleic acids an initial immunization may be performed using an expression vector in the form of naked nucleic acid administered 1M (or SC or ID) in the amounts of 0.5-5 mg at multiple sites. The nucleic acid (0.1 to 1000 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 viais administered at a dose of 5-107 to 5xl09 pfu. 59 For antibodies, a treatment generally involves repeated admi lustration of die aiiti-213P1F11 antibody preparation, via mi acceptable route of administration such as intravenous injection (IV), typically at a dose in die range of about 0.1 to about 10 mg kg body weight. In general, doses in the range of 10-500 mg niAb 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 mg/kg IV of the anti- 213P1F11 niAb preparation represents an acceptable dosing regimen. As appreciated by those of skil l 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, die immunogenicity of a substance, Hie degree of 213P1F11 expression in die patient, the extent of circulating shed 213PIF11 antigen, the desired steady-state concentration level, frequency of treatment, mid the influence of cliemotherapeutic or other agents used in combination with the treatment method of the invention, as well as the health status of a particular patient. Non-limiting preferred human unit doses are, for example, 500μ - lmg, Img - 50nig, 50mg - lOOmg, lOOmg - 200ing, 200mg -300mg, 400mg - SOOnig, 500mg - 600mg, 600mg - 700mg, 700mg - SOOnig, 800mg - 900mg, 900mg - lg, or lmg - 700mg. In certain embodiments, the dose is in a range of 2-5 mg/kg body weight, e.g., with follow on weekly doses of 1-3 mg/kg; 0.5mg, 1, 2, 3, 4, 5, 6, 7, 8, 9, lOmg/kg body weight followed, e.g., in two, three or four weeks by weekly doses; 0.5 - lOmg/kg body weight, e.g., followed in two, tliree or four weeks by weekly doses; 225, 250, 275, 300, 325, 350, 375, 400mg m2 of body area weekly; I-600mg m2 of body area weekly; 225-400mg m2 of body area weekly; these does can be followed by weekly 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, including the sequence of the polynucleotide, molecular weight of the polynucleotide and route of adniinistration. 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 m d the like. Generally, for a polynucleotide 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 mi 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 polynucleotide compared to more direct application to the nucleotide to diseased tissue, as do polynucleotides of increasing length.

In one embodiment, human unit dose forms of T-cells comprise 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 lO cells to about 106 cells, about It)6 cells to about 60 10s cells, about 10s to about lO11 cells, or about 10s to about 5 x 1010 cells. A dose may also about 106 cells/m2 to about 1010 cells/m2, or about 106 cells/m2 to about 10s cells/m2 .

Proteins(s) of tire invention, and/or nucleic acids encoding (lie protein(s), can also be administered via liposomes, which may also serve to: 1) target tlie proleins(s) to a particular (issue, such as lymphoid tissue; 2) to target selectively to diseases cells; or, 3) to increase (lie half-life of the peptide composition. Liposomes include emulsions, foams, micelles, insoluble monolayers, liquid crystals, phospholipid dispersions, lamellar layers and tlie like. In these preparations, tire 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 monoclonal antibodies which bind to tlie CD45 antigen, or with other therapeutic or immunogenic compositions. Tims, liposomes either filled or decorated with a desired peptide of tl e invention can be directed to tlie site of lymphoid cells, where tlie liposomes then deliver tlie peptide compositions. Liposomes for use in accordance with tlie invention are formed from standard vesicle-forming lipids, which generally include neutral and negatively charged phospholipids and a sterol, such as cholesterol. The selection of lipids is generally guided by consideration of, e.g., liposome size, acid lability and stability of the liposomes in the blood stream. A variety of methods are available for preparing liposomes, as described in, e.g., Szoka, el 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 tlie immune system, a ligand to be incorporated into the liposome can include, e.g., antibodies or fragments thereof specific for cell surface determinants of the desired immune system cells. A liposome suspension containing a peptide may be administered intravenously, locally, topically, etc. in a dose which varies according to, inter alia, tlie manner of administration, tlie peptide being delivered, and tlie stage of the disease being treated.

For solid compositions, conventional nontoxic solid carriers may be used which include, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, cellulose, glucose, sucrose, magnesium carbonate, and (lie like. For oral administration, a pharmaceutically acceptable nontoxic composition is formed by incorporating any of the normally employed excipients, such as those carriers previously listed, and generally 10-95% of active ingredient, that is, one or more peptides of tlie 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 m d propellant. Typical percentages of peptides are about 0.01%-20% by weight, preferably about 1 %-10%. The surfactant must, of course, be nontoxic, and preferably soluble in tlie propellant. Representati ve of such agents are tlie esters or partial esters of fatty acids containing from about 6 to 22 carbon atoms, such as caproic, octanoic, lauric, palmitic, stearic, linoleic, linolenic, olesteric and oleic acids 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. l%-20% by weight of the composition, preferably about 0.25-5%. The balance of tlie composition is ordinarily propellant. A carrier can also be included, as desired, as with, e.g., lecithin for intranasal delivery.

XL) Diagnostic and Prognostic Embodiments of 213P1 F11. 1 As disclosed herein, 213P1F11 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, e.g., 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 213P1F1 1 in Normal Tissues and Patient Specimens"). 213P1F11 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, e.g., Merrill et al., J. Urol. 163(2): 503-5120 (2000); Polascik et al., 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 Minimoto et al., Cancer Detect Prev 2000;24(1): 1-12). Therefore, this disclosure of 213P1F11 polynucleotides and polypeptides (as well as 213P1F11 polynucleotide probes and anti-213PlFl l antibodies used to identify the presence of these molecules) and their properties allows skilled artisans to utilize these molecules in metliods 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 metliods which utilize die 213P1F11 polynucleotides, polypeptides, reactive T cells and antibodies are analogous to those metliods from well-established diagnostic assays which employ, e.g., PSA polynucleotides, polypeptides, reactive T cells and antibodies. For example, just as PSA polynucleotides are used as probes (for example in Northern analysis, see, e.g., Sharief et al., Biochem. Mol. Biol. Int. 33(3):567-74(1994)) and primers (for example in PCR analysis, see, e.g., Okegawa et al , J. Urol. 163(4): 1189-1190 (2000)) to observe the presence and/or the level of PSA mRNAs in methods of monitoring PSA overexpression or the metastasis of prostate cancers, the 213P1 F1 1 polynucleotides described herein can be utilized in the same way to detect 213P1F1 1 overexpression or the metastasis of prostate mid oilier cancers expressing this gene. Alternatively, 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 metliods to monitor PSA protein overexpression (see, e.g., Stephan et al, Urology 55(4):560-3 (2000)) or the metastasis of prostate cells (see, e.g., Alaiien et al., Pathol. Res. Pract. 192(3):233-7 (1996)), the 213P1F11 polypeptides described herein can be utilized to generate antibodies for use in detecting 213P1F11 overexpression or the metastasis of prostate cells and cells of other cancers expressing this gene.

Specifically, because metastases involves the movement of cancer cells from an organ of origin (such as d e 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 213P1F1 1 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 213PlFl l-expressing cells (lymph node) is found to contain 213P1F11-expressing cells such as the 213P1F1 1 expression seen in LAPC4 and LAPC9, xenografts isolated from lymph node and bone metastasis, respectively, this finding is indicative of metastasis.

Alternatively 213P1F11 polynucleotides and/or polypeptides can be used to provide evidence of cancer, for example, when cells in a biological sample that do not normally express 213P1F11 or express 213P1F11 at a different level are found to express 213P1F11 or have an increased expression of 213P1F11 62 (see, e.g., the 213P1F1 1 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 tire biological sample for the presence of a second tissue restricted marker (in addition to 213P1F11) such as PSA, PSCA etc. (see, e.g., Alanen et al., Pathol. Res. Pract. 192(3): 233-237 (1996)).

Just as PSA polynucleotide fragments and polynucleotide variants are employed by skilled artisans for use in methods of monitoring PSA, 2 I 3P1FI 1 polynucleotide fragments and polynucleotide variants are used in an analogous maimer. In particular, typical PSA polynucleotides used in methods of monitoring PSA are probes or primers which consist of fragments of the PSA cDNA sequence. Illustrating this, primers used to PCR amplify a PSA polynucleotide must include less than die whole PSA sequence to function in die polymerase chain reaction. In the context of such PCR reactions, skilled artisans generally create a variety of different polynucleotide fragments that can be used as primers in order to amplify different portions of a polynucleotide of interest or to optimize amplification reactions (see, e.g., Caetano-Anolles, G. Biotechniques 25(3): 472-476, 478-480 (1998); Robertson et at. , Mediods Mol. Biol. 98: 12 1 - 154 (1998)). An additional illustration of tire use of such fragments is provided in die Example entitled "Expression Analysis of 213P1F1 1 in Normal Tissues and Patient Specimens," where a 213P 1F11 polynucleotide fragment is used as a probe to show the expression of 213P1F11 RNAs in cancer cells. In addition, variant polynucleotide sequences are typically used as primers and probes for die corresponding niRNAs in PCR and Northern analyses (see, e.g., Sawai et at., Fetal Diagn. Ther. 1996 Nov-Dec 1 1(6):407-13 and Current Protocols In Molecular Biology, Volume 2, Unit 2, Frederick M. Ausubel et al. eds., 1995)). Polynucleotide fragments and variants are useful in this context where they a re capable of binding to a target polynucleotide sequence (e.g., a 213P1F11 polynucleotide shown in Figure 2 or variant diereof) under conditions of high stringency.

Furthermore, PSA polypeptides which contain an epitope dial can be recognized by an antibody or T cell diat specifically binds to that epitope are used in mediods of monitoring PSA. 213P1F11 polypeptide fragments mid polypeptide analogs or variants can also be used in an analogous maimer. This practice of using polypeptide fragments or polypeptide variants to generate antibodies (such as anti-PSA antibodies or T cells) is typical in die art widi a wide variety of systems such as fusion proteins being used by practitioners (see, e.g., Current Protocols In Molecular Biology, Volume 2, Unit 16, Frederick M. Ausubel et al. eds., 1995). In tiiis 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, e.g., 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 die 213P1F1 1 biological motifs discussed herein or a motif-bearing subsequence which is readily identified by one of skill in the art based on motifs available in die art.

Polypeptide fragments, variants or analogs are typically useful in diis context as long as they comprise an epitope capable of generating an antibody or T cell specific for a target polypeptide sequence (e.g. a 213PIF1 1 polypeptide shown in Figure 3).

As shown herein, die 213P1F11 polynucleotides and polypeptides (as well as die 213P1F11 polynucleotide probes and anti-213PlFl 1 antibodies or T cells used to identify the presence of these molecules) exhibit specific properties that make Uiem useful in diagnosing cancers such as Uiose listed in Table I. Diagnostic assays that measure die presence of 213P1F11 gene products, in order to evaluate the 63 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 successfully with PSA. Moreover, these materials satisfy a need in the art for molecules having similar or complementary characteristics 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, e.g., Alanen et ai , Pathol. Res. Pract. 192(3): 233-237 (1996)), and consequently, materials such as 213P 1F11 polynucleotides and polypeptides (as well as the 213P1F1 1 polynucleotide probes and anti-213PlFl 1 antibodies used to identify the presence of these molecules) need to be employed to confirm a metastases of prostatic origin.

Finally, in addition to their use in diagnostic assays, the 213P1F11 polynucleotides disclosed herein have a number of other utilities such as their use in tire identification of oncogenetic associated chromosomal abnormalities in the chromosomal region to which the 213P1F1 1 gene maps (see the Example entitled "Chromosomal Mapping of 213P1F11" below). Moreover, in addition to their use in diagnostic assays, the 213PlFl l-related proteins and polynucleotides disclosed herein have other utilities such as their use in the forensic analysis of tissues of unknown origin (see, e.g., Takahama K Forensic Sci Jul 1996 Jim 28;S0(l -2): 63-9).

Additionally, 213P1F11 -related proteins or polynucleotides of the invention can be used to treat a pathologic condition characterized by the over-expression of 213P1F1 1. For example, the amino acid or nucleic acid sequence of Figure 2 or Figure 3, or fragments of .either, can be used to generate an immune response to a 213P1F11 antigen. Antibodies or other molecules that react with 213P1F 11 can be used to modulate the function of this molecule, mid thereby provide a therapeutic benefit. ' XII.) Inhibition of 213P1F11 Protein Function The invention includes various methods and compositions for inhibiting the binding of 213P1F11 to its binding partner or its association with other prolein(s) as well as methods for inhibiting 213P1F1 1 function.

XJI.A.) inhibition of 213P1F11 With Intracellular Antibodies In one approach, a recombinant vector that encodes single chain antibodies that specifically bind to 213P1F1 1 are introduced into 213P1F1 1 expressing cells via gene transfer technologies. Accordingly, the encoded single chain anti-213P lFH antibody is expressed intracellularly, binds to 213P1F11 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 tecluiology 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 cell surface receptors (see, e.g., Richardson el ai, 1995, Proc. Natl. Acad. Sci. USA 92 : 3137-3141 ; Beerli et ai, 1994, J. Biol. Chem. 289: 23931 -23936; Deshane et ai, 1994, Gene Ther. 1 : 332-337).

Single chain antibodies comprise the variable domains of the heavy and light chain joined by a flexible linker 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 polynucleotide vectors encoding such single 64 chain antibodies in order to precisely target the intrabody to the desired intracellular compartment, For example, intrabodies targeted to tlie endoplasmic reticulum (ER) are engineered to 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 tlie nucleus are engineered to include a nuclear localization signal. Lipid moieties are joined to intrabodies in order to tether tlie intrabody to the cytosolic side of the plasma membrane.

Intrabodies can also be targeted to exert function in tlie cytosol. For example, cytosolic intrabodies are used to sequester factors within the cytosol, thereby preventing them from being transported to their natural cellular destination.

In one embodiment, intrabodies are used to capture 213P1F11 in tlie nucleus, thereby preventing its activity within tl e nucleus. Nuclear targeting signals are engineered into such 213P1F1 1 intrabodies in order to achieve the desired targeting. Such 213P1F11 intrabodies are designed to bind specifically to a particular 213P1F11 domain. In another embodiment, cytosolic intrabodies that specifically bind to a 213P1F11 protein are used to prevent 213P1F1 1 from gaining access to the nucleus, thereby preventing it from exerting any biological activity within tlie nucleus (e.g., preventing 213P1F1 1 from forming transcription complexes with other factors).

In order to specifically direct tlie expression of such intrabodies to particular cells, tlie transcription of the intrabody is placed under tlie regulatory control of an appropriate tumor-specific promoter and/or enhancer. In order to target intrabody expression specifically to prostate, for example, tlie 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 213P1F11 with Recombinant Proteins In another approach, recombinant molecules bind to 213P1F11 and thereby inhibit 213P1F11 function. For example, these recombinant molecules prevent or inl ibit 213P1F11 from accessing/binding to its binding partner(s) or associating with other protein(s). Such recombinant molecules can, for example, contain tlie reactive part(s) of a 213P1F11 specific antibody molecule. In a particular embodiment, tlie 213P1F11 binding domain of a 213P1F11 binding partner is engineered into a dimeric fusion protein, whereby tlie fusion protein comprises two 213P1F11 ligand binding domains linked to tlie Fc portion of a human IgG, such as human IgGl. Such IgG portion can contain, for example, tlie CH2.and CH3 domains and tlie lunge region, but not tlie CH1 domain. Such dimeric fusion proteins are administered in soluble form to patients suffering from a cancer associated with die expression of 213P1F11, whereby die dimeric fusion protein specifically binds to 213P1F11 and blocks 213P1F11 interaction with a binding partner. Such dimeric fusion proteins are furdier combined into niultimeric proteins using known antibody linking technologies.

XII.C.) Inhibition of 213P1F11 Transcription or Translation The present invention also comprises various meU ods and compositions for inhibiting the transcription of die 213P1F1 1 gene. Similarly, die invention also provides methods and compositions for inhibiting die translation of 213P1F1 1 mRNA into protein.

In one approach, a method of inhibiting the transcription of the 213P1F11 gene comprises contacting d e 213P1F11 gene with a 213P1F11 antisense polynucleotide. In another approach, a meUiod of inhibiting 213P1F1 1 mRNA translation comprises contacting a 213P1F11 mRNA with an antisense polynucleotide. In another approach, a 213P1F1 1 specific ribozyme is used to cleave a 213P1F1 1 message, thereby inlvibiting translation. Such antisense and ribozyme based methods can also be directed to d e regulatory regions of the 65 213P1F1 1 gene, such as 213PIF11 promoter and/or enhancer elements. Similarly, proteins capable of inhibiting a 213P1F1 1 gene transcription factor are used to inhibit 213P1F11 mRNA transcription. The various polynucleotides and compositions useful in the aforementioned methods have been described above. The use of antisense and ribozyme molecules to inhibit transcription and translation is well known in the art.

Odver factors mat inhibit the transcription of 213P1F1 1 by interfering with 213P1F 11 transcriptional activation are also useful to treat cancers expressing 213P1F11. Similarly, factors thai interfere with 213P1F11 processing are useful to treat cancers that express 213P1F11. Cancer treatment methods utilizing such factors are also within the scope of the invention.

X1I.D.) General Considerations or Therapeutic Strategies Gene transfer and gene therapy technologies can be used to deliver therapeutic polynucleotide molecules to tumor cells synthesizing 213P1F11 (i.e., antisense, ribozyme, polynucleotides encoding intrabodies and other 213P1 Fl 1 inhibitory molecules). A number of gene therapy approaches are known in the art. Recombinant vectors encoding 213P1F1 1 antisense polynucleotides, ribozynies, factors capable of interfering with 213P1F11 transcription, and so forth, can be delivered to target tumor cells using such gene therapy approaches.

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 die toxicity of the chemotherapeutic agent well.

The anti-tumor activity of a particular composition (e.g., antisense, ribozyme, intrabody), or a combination of such compositions, can be evaluated using various in vitro and in vivo assay systems. In vitro assays that evaluate therapeutic activity include cell growth assays, soft agar assays and other assays indicative of tumor promoting activity, binding assays capable of determining the extent to winch a therapeutic composition will inhibit the binding of 213P1F11 to a binding partner, etc.

In vivo, the effect of a 213P1F11 therapeutic composition can be evaluated in a suitable animal model. For example, xenogenic prostate cancer models can be used, wherein human prostate cancer explains or passaged xenograft tissues are introduced into immune compromised animals, such as nude or SOD mice (Klein et αί , 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 development of primary tumors, micrometastasis, and tire formation of osteoblastic metastases characteristic of late stage disease. Efficacy can be predicted using assays that measure inlubition 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 apoplotic foci and compared to untreated control xenograft-bearing mice. The extent to which apoplotic foci are found in Hie 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 phannaceutical compositions comprising a carrier suitable for the desired deliver)' method. Suitable carriers ■ include any material that when combined with the therapeutic composition retains die anti-tumor function of the therapeutic composition and is generally non-reactive with die patient's immune system. Examples 66 include, but are not limited to, any of a number of standard pharmaceutical carriers such as sterile phosphate buffered saline solutions, bacteriostatic water, and tlie like (see, generally, Remington's Pharmaceutical Sciences 16ϋι Edition, A. Osal., Ed., 1 80).

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 include, but are not limited to, intravenous, parenteral, intraperitoneal, intramuscular, intratumor, intradermal, intraorgan, orthotopic, and tlie like. A preferred formulation for intravenous injection comprises tlie 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 tlie treatment of cancers using tlie foregoing methods will vary with tl e method and tlie target cancer, and will generally depend on a number of other factors appreciated in tlie art.

XIII.) Kits For use in tlie diagnostic and therapeutic applications described herein, kits are also within tile scope of the invention. Such kits can comprise a carrier, package or container that is compartmentalized to receive one or more containers such as vials, tubes, and tlie like, each of the container(s) comprising one of the separate elements to be used in tlie.metliod. For example, tlie container(s) can comprise a probe that is or can be detectably labeled. Such probe can be an antibody or polynucleotide specific for a 213P1 Fl 1-related protein or a 213P1F11 gene or message, respectively. Where tlie method utilizes nucleic acid hybridization to detect the target nucleic acid, tlie kit can also have containers containing nucleotide(s) for amplification of the target nucleic acid sequence and/or a container comprising a reporter-means, such as a biotin-binding protein, such as avidin or streptavidin, bound to a reporter molecule, such as an enzymatic, florescent, or radioisotope label. The kit can include all or part of the amino acid sequence of Figure 2 or Figure 3 or analogs thereof, or a nucleic acid molecules that encodes such amino acid sequences.

The kit of tlie. invention will typically comprise tlie container described above mid one or more other containers comprising materials desirable from a commercial and iiser standpoint, including buffers, diluents, filters, needles, syringes, and package inserts with instructions for use.

A label can be present on tlie container to indicate that tlie composition is used for a specific therapy or non-therapeutic application, and can also indicate directions for either in vivo or in vitro use, such as those described above. Directions and or other information can also be included on an insert which is included with tlie kit.

EXAMPLES: Various aspects of the invention are further described and illustrated by way of the several examples that follow, none of which are intended to limit tlie scope of the invention. 67 Example 1: SSH-Gcncratetl Isolation of a cDNA Fragment of the 2 I 3P 1 PH Gene <.

To isolate genes tliat are over-expressed in bladder cancer, Suppression Subtractive Hybridization (SSH) procedure using cDNA derived from bladder cancer tissues was performed, including invasive transitional cell carcinoma. The 213P1F11 SSH cDNA sequence was derived from a bladder cancer pool minus cDNAs derived from 9 normal tissues. The 213P1F11 cDNA was identified as highly expressed in tire bladder cancer tissue pool, with no expression detected in normal tissues.

The SSH DNA sequence of 166 bp (Figure 1) did not show homology to any known gene. 213P1F1 lv.1 of 3336 bp was identified and the open reading frame cloned from bladder cancer cDNA, revealing an ORF of 242 amino acids (Figure 2 and Figure 3). Other variants of 213P1F11, were also identified and these are listed in Figures 2 and 3. 213P1F11 v. l reveals 100% identity to caspase-14 precursor apoptosis-related cysteine protease protein (Figure 4).

Materials and Methods Human Tissues: The patient cancer and normal tissues were purchased from different sources such as d e NDRI (Philadelphia, PA). mRNA for some normal tissues were purchased from Clontech, Palo Alto, CA.

RNA Isolation: Tissues were homogenized in Trizol reagent (Life Teclmoiogies, Gibco BRL) using 10 ml/ g tissue isolate total RNA. Poly A RNA was purified from total RNA using Qiagen's Oligotex mRNA Mini mid Midi kits. Total and inRNA were quantified by spectropliotonietric analysis (O.D. 260/280 nm) and analyzed by gel electrophoresis.

Oligonucleotides: The following HPLC purified oligonucleotides were used.

DPNCDN (cDNA synthesis primer): ' TT TGATC A AGCTT303 ' (SEQ ID NO: 42) Adaptor 1: 'CTAATACGACTCACTATAGGGCTCGAGCGGGCGCCCGGGCAG3 ' (SEQ ID NO: 43) S 'GGCCCGTCCTAGS 1 (SEQ ID NO: 44) Adaptor 2: 'GTAATACGACTCACTATAGGGCAGCGTGGTCGCGGCCGAG3 ' (SEQ ID NO: 45) 3 'CGGCTCCTAG5 ' (SEQ ID NO: 46) PCR primer 1 : 'CTAATACGACTCACTATAGGGC3 ' (SEQ ID NO: 47) Nested primer (NP)1 : 68 CGAGCGGCCGCCCGGGCAGGA3 ' (SEQ ID NO: 48) Nested primer (NP)2 : 'AGCGTGGTCGCGGCCGAGGA3 ' (SEQ ID NO: 49) Suppression Sub ractive Hybridization: Suppression Subtractive Hybridization (SSH) was used to identify cDNAs corresponding to genes that -may be differentially expressed in bladder cancer. The SSH reaction utilized cDNA from bladder cancer and normal tissues.

The gene 213P1F1 1 sequence was derived from a bladder cancer pool minus normal tissue cDNA subtraction. The SSH DNA sequence (Figure 1) was identified.

The cDNA derived from of pool of normal tissues was used as the source of the "driver" cDNA, while the cDNA from a pool of bladder cancer tissues was used as the source of the "tester" cDNA. Double stranded cDNAs corresponding to tester and driver cDNAs were synthesized from 2 ¾ of poly(A)+ RNA isolated from the relevant xenograft tissue, as described above, using CLONTECH's PCR-Select cDNA Subtraction Kit and 1 ng of oligonucleotide DPNCDN as primer. First- and second-strand synthesis were carried out as described in the Kit's user manual protocol (CLONTECH Protocol No. PT1117-1, Catalog No. Κ1804-Ί). The resulting cDNA was digested widi Dpn II for 3 hrs at 37°C. Digested cDNA was extracted with phenol/chloroform (1 : 1) and ethanol precipitated.

Driver cDNA was generated by combining in a 1 : 1 ratio Dpn II digested cDNA from the relevant tissue source (see above) with a mix of digested cDNAs derived from the nine normal tissues: stomach, skeletal muscle, lung, brain, liver, kidney, pancreas, small intestine, and heart. ' Tester cDNA was generated by diluting 1 μΐ of Dpn II digested cDNA from the relevant tissue source (see above) (400 ng) in 5 μΐ of water. The diluted cDNA (2 μΐ, 160 ng) was dien ligated to 2 μΐ of Adaptor 1 and Adaptor 2 (10 μ ), in separate ligation reactions, in a total volume of 10 μΐ at 16°C overnight, using 400 11 of T4 DNA ligase (CLONTECH). Ligation was terminated with 1 μΐ of 0.2 M EDTA and heating at 72°C for 5 min.

The first hybridization was performed by adding 1.5 μΐ (600 ng) of driver cDNA to each of two tubes containing 1.5 μΐ (20 ng) Adaptor 1 - and Adaptor 2- ligated tester cDNA. In a final volume of 4 μΐ, the samples were overlaid with mineral oil, denatured in an MJ Research thermal cycler at 98°C for 1.5 minutes, and then were allowed to hybridize for 8 hrs at 68°C. The two hybridizations were then mixed together with an additional 1 μΐ of fresh denatured driver cDNA and were allowed to hybridize overnight at 68°C. The second hybridization was then diluted in 200 μΐ of 20 niM Hepes, pH 8.3, 50 niM NaCl, 0.2 niM EDTA, heated at 70°C for 7 min. and stored at -20°C.

PCR Amplification, Cloning and Sequencing of Gene Fragments Generated from SSH: To amplify gene fragments resulting from SSH reactions, two PCR amplifications were performed. In die primary PCR reaction 1 μΐ of die diluted final hybridization mix was added to 1 μΐ of PCR primer 1 (10 μ ), 0.5 μΐ dNTP mix (10 μΜ), 2.5 μΐ 10 x reaction buffer (CLONTECH) mid 0.5 μΐ 50 x Advantage cDNA polymerase Mix (CLONTECH) in a final volume of 25 μΐ. PCR 1 was conducted using the^following 69 conditions: 75°C for 5 min., 94°C for 25 sec, then 27 cycles of 94°C for 10 sec, 66°C for 30 sec, 72°C 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 μΐ 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 μΜ) were used instead of PCR primer 1. PCR 2 was performed using 10-12 cycles of 94°C for 10 sec, 68°C for 30 sec, and 72°C for 1.5 minutes. The PCR products were analyzed using 2% agarose gel electrophoresis.

The PCR products were inserted into pCR2.1 using the T/A vector cloning kit (Invitrogen).

Transformed E. coli were subjected to blue/white and ampicillin selection. White colonies were picked and arrayed into 96 well plates and were grown in liquid culture overnight. To identify inserts, PCR amplification was performed on 1 ml of bacteri al culture using the conditions of PCR1 and NP1 mid NP2 as primers. PCR products were analyzed using 2% agarose gel electrophoresis.

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

RT-PCR Expression Analysis: First strand cDNAs can be generated from 1 ^ig of iiiRNA 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 37°C for 20 min. After completing the reaction, tire volume can be increased to 200 μΐ with water prior to normalization. First strand cD As from 16 different normal human tissues can be obtained from Clontech.

Normalization of the first strand cDNAs from multiple tissues was performed by using the primers 5'atatcgccgcgctcgtcgtcgaoaa3 ' (SEQ ID NO: 50) and 5 'agccacacgcagctcattgtagaagg 3 ' (SEQ ID NO: 51) to amplify β-actin. First strand cDNA (5 μΐ) were amplified in a total volume of 50 μΐ containing 0.4 μΜ primers, 0.2 μΜ each dNTPs, 1XPCR buffer (Clontech, 10 niM Tris-HCL, 1.5 niM MgCl2, 50 niM KC1, pH8.3) and IX Klentaq DNA polymerase (Clontech). Five μΐ 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 94°C for 15 sec, followed by a 18, 20, and 22 cycles of 94°C for 15, 65°C for 2 min, 72°C for 5 sec. A final extension at 72°C was carried out for 2 min. After agarose gel electrophoresis, die band intensities of the 283 b.p. β-actin bands from multiple tissues were compared by visual inspection. Dilution factors for the first strand cDNAs were calculated to result in equal β-actin band intensities in all tissues after 22 cycles of PCR. Tliree rounds of normalization can be required to acliieve equal band intensities in all tissues after 22 cycles of PCR.

To determine expression levels of the 213P1F11 gene, 5 μΐ 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 tire 213P1F11 SSH sequence and are listed below: 213P1F11.1 '- GGATACCAGGGAACGCTTGGAG - 3 ' (SEQ ID NO: 52) 70 213P1F11.2 '- TTTGACCTTTCCTG CTC AAGT AACC - 3' (SEQ ID NO: 53) ) A typical RT-PCR expression analysis is shown in Figure 14. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC xenograft pool (LAPC-4AD, LAPC-4AI, LAPC-9AD and LAPC-9AI), bladder cancer pool, breast cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to aclin and GAPDH. Semiquantitative PCR, using primers to 213P1 F11, was performed at 26 and 30 cycles of amplification. Results show strong expression of 213P1F11 in bladder cancer pool, breast cancer pool, xenograft pool, and cancer metastasis pool, but not in tlie vital pools.

Example 2: Full Length Cloninti of 213P1F11 The 213P1F11 SSH cDNA sequence was derived from a bladder cancer pool minus normal tissues cDNA subtraction. The SSH cDNA sequence (Figure 1) was designated 213P1F11.

The SSH DNA sequence of 166 bp (Figure 1) did not show homology to any known gene. The full-length cDNA 213P1F11 was cloned from bladder cancer cDNA. Variants of 213P1F11 were identified and these are listed in Figures 2 and 3. 213P1F11 v. l reveals 100% identity to caspase-14 precursor apoptosis-related cysteine protease protein (Figure 4).

Example 3: Chromosomal Manning of 213P1 F11 Chromosomal localization can implicate genes in disease paUiogenesis. Several chromosome mapping approaches are available including fluorescent in situ hybridization (FISH), human/hamster radiation hybrid (RH) panels (Walter et al., 1994; Nature Genetics 7:22; Research Genetics, H ntsville Al), human-rodent somatic cell hybrid panels such as is available from tlie Coriell Institute (Camden, New Jersey), and genomic viewers utilizing BLAST homologies to sequenced and mapped genomic clones (NCBI, Bethesda, Maryland). 213P1F11 maps to chromosome 19 l3.1 using 213P1F11 sequence and tlie NCBI BLAST tool: located at tlie World Wide Web (.ncbi.nlm.nih.gov/geiiome/seq/page.cgi?F=HsBlast.hUnl&(&ORG=Hs).

Example 4: Expression Analysis of 213P1F11 in Normal Tissues and Patient Specimens Expression analysis by RT-PCR demonstrated that 213P 1F11 is strongly expressed in bladder cancer patient specimens (Figure 14). First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC xenograft pool (LAPC-4AD, LAPC-4A1, LAPC-9 D and LAPC-9AI), bladder cancer pool, breast cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 213P 1 F1 1, was performed at 26 and 30 cycles of amplification. Results show strong expression of 213P1FU in bladder cancer pool, breast cancer pool, xenograft pool, and cancer metastasis pool, but not in tlie vital pools.

To determine the relative expression of 213P1F1 1 v. l compared to 213P1FI 1 v.2 in human cancers, primers were designed flanking tlie insertion in 213P 1F11 v.2 (Figure 15). Using these primers, amplification 71 of 213P 1F11 v.1 will generate a PCR fragment of 165 bp, whereas 213P1F1 1 v.2 will generate a PCR fragment of 249 bp as depicted in Figure 15. The PCR product of 165bp will also correspond to tlie variants 213P1F11 v.3, v.4, v.5, v.6 and v.7. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), bladder cancer pool, breast cancer pool, LAPC xenograft pool (LAPC-4AD, LAPC-4AI, LAPC-9AD and LAPC-9A1), and 213P1F11 v. l plasniid control. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers depicted above, was performed at 35 cycles of amplification. Results show strong expression of 213P1F1 1 v. 1 in bladder cancer pool, breast cancer pool, LAPC xenograft pool, and tlie plasmid positive control. A lower expression of the 249 bp 213P1F1 1 v.2 product was delected in breast cancer pool, LAPC xenograft pool, and to lower extent in bladder cancer pool. Altogether these data show that expression of 213P 1F11 v. l is more abundant than 213P1F1 I v.2 in human patient cancer samples.

Extensive northern blot analysis of 213P1F 11 in multiple human normal tissues is shown in Figure 16. Strong expression was only detected in skin tissue. A weak transcript is detected in normal thymus but not in tlie other tissues tested.

RNA was extracted from normal prostate, LAPC-4AD, LAPC-4AI, LAPC- AD and LAPC-9AI prostate cancer xenografts. Northern blot with 10 n of total RNA/lane was probed with 2 J3P1F1 1 SSH sequence (Figure 18). Results show expression of 213P1F11 in tlie LAPC-9AI xenograft, but not in tl e other xenografts nor in normal prostate.

Expression of 213P1F11 in patient bladder cancer specimens is shown in Figure 17. RNA was extracted from normal bladder (N), bladder cancer cell lines (UM-UC-3 and SCaBER), bladder cancer patient tumors (T) and normal tissue adjacent to bladder cancer (NAT). Northern blots with 10 ng of total RNA were probed with tlie 213P1F11 SSH fragment. Size standards in kilobases are indicated on tlie side. Results show strong expression of 213P1F11 in tlie bladder tumor tissues but not in normal bladder, nor in tlie bladder cancer cell lines.

Figure 19 shows that 213P1F1 1 was expressed in breast cancer patient tissues. RNA was extracted from normal breast (N), breast cancer cell lines (DU4475, MCF7 arid CAMA-1), breast cancer patient tumors (T) and breast cancer metastasis to lymph node (Met). Northern blots with 10 tig of total RNA were probed with tlie 213P1F11 SSH fragment. Results show strong expression of 213P1F11 in tl e breast tumor tissues as well as in tlie cancer metastasis specimen. Weak expression was also detected in the CAMA-1 cell line, but not in tlie other 2 breast cancer cell lines tested.

The restricted expression of 213P1F11 in normal tissues and tlie expression detected in bladder cancer, breast cancer, prostate cancer xenograft, and cancer metastases suggest that 213P1F1 1 is a potential therapeutic target and a diagnostic marker for human cancers.

Example 5: Transcript Variants of 213P1F11 Transcript variants are variants of matured mRNA from tlie same gene by alternative transcription or alternative splicing. Alternative transcripts are transcripts from tlie same gene but start transcription at different points. Splice variants are mRNA variants spliced differently from tlie 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. 72 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 (5' or 3 ' end) portions, from the original transcript. Transcript variants can code for similar or different proteins with die same or a similar function or may encode proteins with different functions, and may be expressed in the same tissue at the same time, or at different tissue, or at different times, proteins encoded by transcript variants can have similar or different cellular or extracellular localizations, i.e., be secreted.

Transcript variants are identified by a variety of art-accepted methods. For example, alternative transcripts and splice variants are identified in a full-length cloning experiment, or by use of full-length transcript and EST sequences. First, all human ESTs were grouped into clusters which show direct or indirect identity with each other. Second, ESTs in the same cluster were further grouped into sub-clusters mid 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 variant for that gene (see, e.g., located at the World Wide Web (.doubletvvist.com/products/cl l_agentsOverview.jhtml). Even when a variant is identified that is not a full-length clone, that portion of the variant is very useful for antigen generation and for further cloning of the full-length splice variant, using techniques known in the art.

Moreover, computer programs are available in the art that identify transcript variants based on genomic sequences. Genomic-based transcript variant identification programs include FgenesH (A. Salamov and V. Solovyev, "Ab initio gene finding in Drosophila genomic DNA," Genome Research. 2000 April; 10(4):516-22); Grail (http://compbio.ornl.gov/Grail-bin/EmptyGrailFonn) mid GenScan (http://genes.mit.edii/GENSCAN.html). For a general discussion of splice variant identification protocols see., e.g., Southan, C., A genomic perspective on human proteases, FEBS Lett. 2001 Jun 8; 498(2 -3):214-8; de Souza, S.J., et al., Identification of human chromosome 22 transcribed sequences with ORF expressed, sequence tags, Proc. Natl Acad Sci U S A. 2000 Nov 7; 97(23): 12690-3.

To 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, mid 5 ' RACE validation, etc. (see e.g., Proteomic Validation: Brennan, S.O., et al, Albumin banks peninsula: a new termination variant -characterized by electrospray mass spectrometry, Biochem Biophys Acta. 1999 Aug 17; 1433(l-2):321-6; Ferranti P, et al. , Differential splicing of pre-messenger RNA produces multiple forms of mature caprine alpha(sl )-casein, Eur J Biochem. 1997 Oct l ;249( l ): l-7. For PCR-based Validation: Wellinann S. et at., . Specific reverse transcriplion-PCR quantification of vascular endothelial growth factor (VEGF) splice variants by LightCycler teclmology, Clin Chem. 2001 Apr;47(4):654-60; Jia, H.P., et al , Discover,' of new human beta-defensins using a genomics-based approach, Gene. 2001 Jan 24; 263(1-2):211-8. For PCR-based and 5 ' RACE Validation: Brigle, K.E., et al., 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 tire 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 213P1F11 has a particular expression profile related to cancer. Alternative transcripts mid splice variants of 213P1F11 may also be involved in cancers in die same or different tissues, thus serving as tumor-associated markers/antigens. 73 The exon composition of the original transcript, designated as 213P1F11 v.1, is shown in Table XXIIIA. Using the full-length gene and EST sequences, two splice variants were identified, designated as 213P1F11 v.2 and 213P1F11 v.3. Compared with 213P1F11 v. l, splice variant 213P1F11 v.2 had a longer exon 6 while 213P1F11 had a longer exon 5. Using the computer program GenScan, one alternative transcript was identified, designated as 213P1F11 v.4. Tliis alternative transcript had diree different leading exons in place of the first two exons of 213P1F11 v. l . The exon composition of the alternative transcript 213P1F11 v.4 is shown in Table XXIIIB. Since 213P1F11 v.4 shares tire same exons 5 and 6 as 213P1F11 v.1 , splice variants of this alternative transcript with a longer exon 5 or a longer exon 6, or both, may exist in human tissues. In fact, each different combination of exons in spatial order, e.g., exons 1, 2, 3, 4 and 7, is a potential splice variant. Figure 13 shows the schematic alignment of exons of the two transcripts (in addition to variants 2 and 3).

Tables XXIV through XXVII are set forth herein on a variant-by-variant basis. Table XXIV shows the nucleotide sequences of transcript variant 2 through variant 4. Table XXV shows the alignment of transcript variant 2 through variant 4, each with the nucleic acid sequence of 213P1F11 variant 1. Table XXVI lays out amino acid translation of transcript variant 2 throug variant 4 for die identified reading frame orientation. Table XXVII displays alignments of the amino acid sequences encoded by splice variant 2 tliroug variant 4, each with that of 213P1F1 1 variant 1. Table XXVIII displays clustal alignments of 213PIF11 protein variant 1 tlirough variant 6.

Example >: Sinule Nucleotide Polymorphisms of 213P1F11 Single Nucleotide Polymorphism (SNP) is a single base pair variation in nucleotide sequences. At a specific point of the genome, there are four possible nucleotide base pairs: A/T, C/G, G/C and T/A. Genotype refers to tire base pair make-up of one or more spots in the genome of an individual, while haplotype refers to base pair make-up of more than one varied spots on tire same DNA molecule (chromosome in higher organism). SNPs that occur on a cDNA are called cSNPs. These cSNPs may change amino acids of the protein encoded by the gene and thus change d e functions of the protein. Some SNPs cause inherited diseases and some others contribute to quantitative variations in phenotype and reactions to environmental factors including diet and drugs among individuals. Therefore, SNPs and/or combinations of alleles (called haplolypes) have many applications including diagnosis of inherited diseases, delemiinalion of drug reactions and dosage, identification of genes responsible for disearses and discovery of genetic relationship between individuals (P. Nowotny, J. M. Kwon and A. M. Goate, " SNP analysis to dissect human trails," 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 Jim; 22(6):298-305; J. H. Riley, C. J. Allan, E. Lai and A. Roses, " The use of single nucleotide polymorphisms in die isolation of common disease genes," Pharmacogenomics. 2000 Feb; l ( l):39-47; R. Judson, J. C. Stephens and A. Windemutli, "The predictive power of haplolypes in clinical response," Pharmacogenomics. 2000 feb; 1(1): 15-26). ' SNPs are identified by a variety of art-accepted methods (P. Bean, "The promising voyage of SNP target discovery," Am. Clin. Lab. 2001 Ocl-Nov; 20(9): 18-20; K. M. Weiss, "In search of human variation," Genome Res. 1998 Jul; 8(7):691 -697; M. M. She, "Enabling large-scale pharmacogenetic studies by high-throughput mutation detection and genotyping technologies," Clin. Chein. 2001 Feb; 47(2): 164-172). For 74 example, SNPs are 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 tlie rapid accumulation of sequence data in public and private databases, one can discover SNPs by comparing sequences using computer programs (Z. Gu, L. Hillier and P. Y. vvok, "Single nucleotide polymorphism hunting in cyberspace," Hum. Mutat. 1998; 12(4):221-225). SNPs 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 (P. 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. Envin, P. Grass, B. Hines and A. Duesterhoeft, "High-throughput SNP genotyping with tlie Masscode system," Mol. Diagi . 2000 Dec; 5(4):329-340).

Using the methods described above, six SNPs were identified in the original transcript, 213P1 F1 1 v. l, at positions 473 (T/C), 737 (C/A), 2027 (C/T), 2037 (T/C), 2268 (A/G) and 3196 (A T). The transcripts or proteins with alternative alleles were designated as variants 213P1F1 1 v.5, v.6, v.7, v.8, v.9, and v. lO. Figure 10 shows the schematic alignment of the nucleotide variants. Figure 11 shows the schematic alignment of protein variants, corresponding to nucleotide variants. Nucleotide variants that code for the same amino acid sequence as variant 1 are not shown in Figure 11. These alleles of the SNPs, though shown separately here, can occur in different combinations (haplotypes) and in any one of the transcript variants that contains the sequence context of tlie SNPs , e.g., 213P1F11 v.2, 213P1F11 v.3 or 213Plfl l v.4.

Example 7: Production of Recombinant 213P1F11 in Pi okaryotic Systems To express recombinant 213P1F11 and 213P1F11 variants in prokaryotic cells, tlie full or partial lengdi 213P1F11 and 213P1F11 variant cDNA sequences are cloned into any one of a variety of expression vectors known in tl e art. One or more of tlie following regions of 213P1F11 or 213P1FU variants are expressed in these constructs, amino acids 1 to 242 of 213P1F 1 1 variant 1 , amino acids 1 -230 of variant 2, ammo acids 1 -146 of variant 3, amino acids 1 -321 of variant 4; 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 213P1F1 1, variants, or analogs thereof.

A. In vitro transcription and translation constructs: pCRII: To generate 213P1F11 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 213P1F11 cDNA. The pCRII vector has Sp6 and T7 promoters flanking tlie insert to drive tlie transcription of 213P1F11 RNA for use as probes in RNA in situ hybridization experiments. These probes are used to analyze the cell and tissue expression of 213P IF 11 at tlie RNA level. Transcribed 213P1F11 RNA representing tlie cDNA amino acid coding region of tlie 213P1F11 gene is used in in vitro translation systems such as the TnT1 M Coupled Reticulolysate Sytem (Promega, Corp., Madison, WI) to synthesize 213P1F11 protein.

B. Bacterial Constructs: 75 pGEX Constructs: To generate recombinant 213P1F11 proteins in bacteria that are fused to the Glutathione S-transferase (GST) protein, all or parts of the T- fusion vector of the pGEX family (Amersham Pharmacia Biotech, Piscatavvay, NJ). These constructs allow controlled expression of recombinant 213P1F11 protein sequences with GST fused at the aniino-lerminus 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 die 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, e.g., of the open reading frame (ORF). A proteolytic cleavage site, such as the PreScission1M recognition site in pGEX-6P-l, may be employed such dial it permits cleavage of the GST tag from 213P1F11 -related protein. The ampicillin resistance gene and pBR322 origin permits selection and maintenance of the pGEX plasmids in E. coli. pMAL Constructs: To generate, in bacteria, recombinant 213P1F11 proteins that are fused to maltose-binding protein (MBP), all or parts of the 213P1F11 cDNA protein coding sequence are fused to the MBP gene by cloning into the pMAL-c2X and pMAL-p2X vectors (New England Biolabs, Beverly, MA). These constructs allow controlled expression of recombinant 213P1F1 1 protein sequences with MBP fused at the amino-tenninus and a 6X His epitope tag at the carboxyl-terminus. The MBP mid 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 213P1F11. The pMAL-c2X mid 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. pET Constructs: To express 213P1F1 1 in bacterial cells, all or parts of Hie 213P1F1 1 cDNA protein coding sequence are cloned into die pET family of vectors (Novagen, Madison, WI). These vectors allow tightly controlled expression of recombinant 213P1F11 protein in bacteria with mid without fusion to proteins that enhance solubility, such as NusA and tliioredoxin (Trx), and epitope tags, such as 6X His and S-Tag™ dial aid purification mid detection of the recombinant protein. For example, constructs are made utilizing pET NusA fusion system 43.1 such that regions of the 213P1F11 protein are expressed as amino-terminal fusions to NusA.

C. Yeast Constructs: . pESC Constructs: To express 213P1F1 1 in (lie yeast species Sacchctromyces cerevisiae for generation of recombinant protein and functional studies, all or parts of the 213P 1F1 1 cDNA protein coding sequence are cloned into the pESC family of vectors each of which contain 1 of 4 selectable markers, H1S3, TRPl, LEU2, and URA3 (Stratagene, La Jolla, CA). These vectors allow controlled expression from die same plasnud of up to 2 different genes or cloned sequences containing eidier Flag™ or Myc epitope tags in the same yeast cell. This system is useful to confirm protein-protein interactions of 213P1F11. 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 213P1F11 in the yeast species Saccharornyces po be, all or parts of the 213P 1F11 cDNA protein coding sequence are cloned into the pESP family of vectors. These vectors 76 allow controlled high level of expression of a 213P1F11 protein sequence tliat is fused at either tlie amino terminus or. at tlie carboxyl terminus to GST which ids purification of die recombinant protein. A Flag™ epitope tag allows detection of the recombinant protein with anti- Flag™ antibody.

Example 8: Production of Recombinant 213P1F11 in Eukaryotic Systems A. Mammalian Constructs: To express recombinant 213P1F1 1 in eukaryotic cells, tlie full or partial length 213P1F11 cDNA sequences can be cloned into any one of a variety of expression vectors known in the art. One or more of die following regions of 213P1F1 1 are expressed in these constructs, amino acids 1 to 242, 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 213P1F1 1, variants, or analogs diereof. In certain embodiments a region of a specific variant of 213P1F11 is expressed that encodes an amino acid at a specific position which differs from tlie amino acid of any other variant found at that position. In other embodiments, a region of a variant of 213P1F11 is expressed that lies partly or entirely within a sequence that is unique to tliat variant.

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 tlie anti-213P 1F11 polyclonal serum, described . herein. pcDNA4/HisMax Constructs: To express 213P1F11 in mammalian cells, a 213P1F1 1 ORF, or portions thereof, of 213P1F11 are cloned into pcDNA4/HisMax Version A (Invitrogen, Carlsbad, CA). Protein expression is driven from tlie cytomegalovirus (CMV) promoter and tlie SP16 translational enhancer. The recombinant protein has Xpress™ and six liistidine (6X His) epitopes fused to tlie amino-terminus. The pcDNA4/HisMax vector also contains the bovine growth hormone (BGH) polyadenylation signal and transcription tenmnation sequence to enhance mRNA stability along with tlie S V40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen. Tlie Zeocin resistance gene allows for selection of mammalian cells expressing tlie protein and die ampicillin resistance gene and ColEl origin permits selection and maintenance of the plasmid in it. coli. pcDNA3.1/MvcHis Constructs: To express 213P1F11 in mammalian cells, a 213P1F11 ORF, or portions diereof, of 213P1F11 with a consensus Kozak translation initiation site are cloned into pcDNA3.1/MycHis Version A (Invitrogen, Carlsbad, CA). Protein expression is driven from tlie cytomegalovinis (CMV) promoter. The recombinant proteins have die myc epitope and 6X His epitope fused to the carboxyl-terminus. The pcDNA3.1 MycHis vector also contains tlie bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability, along widi tlie SV40 origin for episomal replication and simple vector rescue in cell lines expressing tlie large T antigen. Tlie Neomycin resistance gene can be used, as it allows for selection of mammalian cells expressing the protein and d e ampicillin resistance gene and ColEl origin permits selection and maintenance of the plasmid in E. coli. pcDNA3.1/CT-GFP-TOPQ Construct: To express 213P1F11 in mammalian cells and to allow detection of the recombinant proteins using fluorescence, a 213P1F11 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 die 77 Green Fluorescent Protein (GFP) fused to tlie carboxyl-terniinus facilitating non-invasive, in vivo detection and cell biology studies. The pcD A3. lCT-GFP-TOPO vector also contains tire 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 tlie large T antigen. The Neomycin resistance gene allows for selection of mammalian cells that express tlie protein, and the ampicillin resistance gene and ColEl origin permits selection and maintenance of the plasmid in E. coli. Additional constructs with an amino-ter inal GFP fusion are made in pcDNA3.1 NT-GFP-TOPO spanning tlie entire lengdi of a 213P1F11 protein.

PAPtag: A 213P1F11 ORF, or portions thereof, is cloned into pAPtag-5 (GenHunter Corp.

Nashville, TN). This construct generates air alkaline phosphatase fusion at tlie carboxyl-termimis of a 213P1F11 protein while fusing tlie IgGic signal sequence to tlie amino-terminiis. Constructs are also generated in which alkaline phosphatase with an aimno-terminal IgGic signal sequence is fused to tlie amino-terminus of a 213P1F1 1 protein. The resulting recombinant 213P1F1 1 proteins are optimized for secretion into tlie media of transfected mammalian cells and can be used to identify proteins such as ligands or receptors 'that interact with 213P1F1 1 proteins. Protein expression is driven from die CMV promoter and tlie recombinant proteins also contain myc and 6X His epitopes fused at die carboxyl-terniinus diat facilitates detection and purification. The Zeocin resistance gene present in tlie vector allows for selection of mammalian cells expressing die recombinant protein mid tlie ampicillin resistance gene permits selection of tlie plasmid in E. coli. ptag5: A 213P1F11 ORF, or portions thereof, is cloned into pTag-5. This vector is similar to pAPtag but without tlie alkaline phosphatase fusion. This construct generates 213P1FJ 1 protein with an amino-terminal IgGx signal sequence and myc and 6X His epitope tags at tlie carboxyl-terniinus diat facilitate detection and affinity purification. The resulting recombinant 213P1F11 protein is optimized for secretion into die media of transfected mammalian cells, and is used as immunogen or ligand to identify proteins such as ligands or receptors diat interact with die 213P1F11 proteins. Protein expression is driven from die CMV promoter. The Zeocin resistance gene present in die vector allows for selection of mammalian cells expressing the protein, and die ampicillin resistance gene permits selection of the plasmid in E. coli.

PsecFc: A 213P1F11 ORF, or portions thereof, is also cloned into psecFc. The psecFc vector was assembled by cloning die human immunoglobulin Gl (IgG) Fc (hinge, CH2, CH3 regions) into pSecTag2 (Iiivitrogen, California). This constnict generates mi IgGl Fc fusion at die carboxyl-terniinus of the 213P 1F1 1 proteins, while fusing die IgGK signal sequence to N-termhiiis. 213P 1F11 fusions utilizing die riiurine IgGl Fc region are also used. The resulting recombinant 213P1F11 proteins are optimized for secretion into die media of transfected mammalian cells, and can be used as immuiiogens or to identify proteins such as ligands or receptors that interact with 213P1F11 protein. Protein expression is driven from die CMV promoter. The Iiygromycin resistance gene present in die vector allows for selection of mammalian cells diat express die recombinant protein, and die ampicillin resistance gene permits selection of the plasmid in E. coli. pSRa Constnicts: To generate mammalian cell lines Uiat express 213P1 F1 1 constitutively, 213P 1F11 ORF, or portions diereof, of 213P 1 F1 1 are cloned into pSRa constructs. Amphotropic and ecotropic retroviruses are generated by transfection of pSRa constructs into die 2 3T- 10A 1 packaging line or 78 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 lines, resulting in the integration of the cloned gene, 213P1F11, 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 tire protein, and tire ampicillin resistance gene and ColEl origin permit selection and maintenance of die plasmid in E. coli. The retroviral vectors can thereafter be used for infection mid 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 FLAG™ tag to the carboxyl-terminus of 213P1F11 sequences to allow detection using anti-Flag antibodies. For example, Hie FLAG™ sequence 5' gat tac aag gat gac gac gat aag 3 ' (SEQ ID NO: 54) 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/όΧ His fusion proteins of the full-length 213P1F11 proteins.

Additional Viral Vectors: Additional constructs are made for viral-mediated delivery and expression of 213P1F11. High virus titer leading to high level expression of 213P1F11 is achieved in viral delivery systems such as adenoviral vectors and herpes a mpl icon vectors. A 213P1F11 coding sequences or fragments thereof are amplified by PCR and subcloned into the AdEasy shuttle vector (Stratagene). Recombination and virus packaging are performed according to the manufacturer's instructions to generate adenoviral vectors. Alternatively, 213P1F1 1 coding sequences or fragments thereof are cloned into 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 213P1F11 in mammalian cells, coding sequences of 213P1F11, or portions thereof, are cloned into regulated manmialian expression systems such as tire T-Rex System (Invitrogen), the GeneSwitch System (Invitrogen) and the tightly-regulated Ecdysone System (Sratagene). These systems allow the study of the temporal mid concentration dependent effects of recombinant 213P1F11. These vectors are thereafter used to control expression of 213P1F1 1 in various cell lines such as PC3, NIH 3T3, 293 or rat-1 cells.

B. Baculovirus Expression Systems To generate recombinant 213P1F11 proteins in a baculovirus expression system, 213P1 F1 1 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-213PlFl l is co-transfected with helper plasmid pBac-N-Blue (Invitrogen) into SF9 (Spocloptera frugiperda) insect cells to generate recombinant baculovirus (see Invitrogen instruction manual for details). Baculovirus is then collected from cell supernatant and purified by plaque assay.

Recombinant 213P1F1 1 protein is then generated by infection of HighFive insect cells (Invitrogen) with purified baculovirus. Recombinant 213PIF11 protein can be detected using anti-213PlFl 1 or anti-His-tag antibody. 213P1F11 protein can be purified and used in various cell-based assays or as immunogen to generate polyclonal and monoclonal antibodies specific for 213P1F11. 79 Example 9: Antigenicity Profiles and Secondary Structure Figure 5A-D, Figure 6A-D, Figure 7 A-D, Figure 8A-D, and Figure 9A-D depict graphically five amino acid profiles of the 213P1F11 variants 1 tlirough 4 respectively, each assessment available by accessing the ProtScale website located at the World Wide Web (.expasy.cli/cgi-bin/prolscale.pi) on the ExPasy molecular biology server.

These profiles: Figure 5, Hydrophilicity, (Hopp T.P., Woods K.R., 1981. Proc. Natl. Acad. Sci. U.S.A. 78:3824-3828); Figure 6, Hydropathicity, (Kyte I, Doolittle R.F., 1982. J. Mol. Biol. 157: 105-132); Figure 7, Percentage Accessible Residues (Janin J., 1979 Nature 277:491-492); Figure 8, Average Flexibility, (Bhaskaran R., and Ponniiswamy P. ., 1988. Int. J. Pept. Protein Res. 32:242-255); Figure 9, Beta-turn (Deleage, G., 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 the 213P1 Fl 1 protein. Each of the above amino acid profiles of 213P1F1 1 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 mid 1.

Hydrophilicity (Figure 5), Hydropathicity (Figure 6) and Percentage Accessible Residues (Figure 7) profiles were used to determine stretches of hydrophilic amino acids (i.e., 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 tire surface of the protein, and thus available for immune recognition, such as by antibodies.

Average Flexibility (Figure 8) and Beta-turn (Figure 9) profiles determine stretches of amino acids (i.e., values greater than 0.5 on the Beta-turn profile and the Average Flexibility profile) that are not constrained in secondary structures 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 213P1F11 protein and of the variant proteins indicated, e.g., by the profiles set forth in Figure 5A-D, Figure 6A-D, Figure 7A-D, Figure 8A-D, and/or Figure 9A-D are used to prepare imnuuiogens, eitlier peptides or nucleic acids that encode them, to generate therapeutic and diagnostic anti-213PlFl l antibodies. The immunogen can be any 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 or more than.50 contiguous amino acids, or the corresponding nucleic acids that encode them, from the 213P1F11 protein variants listed in Figures 2 and 3. In particular, peptide imnuuiogens of the invention can comprise, a peptide region of at least 5 amino acids of Figures 2 and 3 in any whole number increment up to the full length of the respective variant's sequence that includes an amino acid position having a value greater than 0.5 in the Hydroplulicity profiles of Figure 5 A-D; a peptide region of at least 5 amino acids of Figures 2 and 3 in any whole number increment up to die full length of the respective variant's sequence that includes an amino acid position having a value less than 0.5 in the Hydropathicity profile of Figures 6 A-D; a peptide region of at least 5 amino acids of Figures 2 and 3 in any whole number increment up to the full length of the respective variant's sequence that includes an amino acid position having a value greater than 0.5 in the Percent Accessible Residues profiles of Figure 7 A-D; a peptide region of at least 5 amino acids of Figures 2 and 3 in any whole number increment up to the full length of the respective variant's sequence that includes an amino acid position having a value greater than 0.5 in the Average Flexibility profiles on Figure 8 A-D; and, a peptide region of at least 5 amino acids of 80 Figures 2 and 3 in any whole number increment up to the full length of the respective variant's sequence that includes an amino acid position having a value greater than 0.5 in the Beta-turn profile of Figures 9 A-D. Peptide immunogens of the invention can also comprise nucleic acids 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 excipient compatible with human physiology.

The secondary structures of 213P 1F1 ) variants 1 through 4, 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 (Guermeur, 1997, http://pbil.ibcp.fr/cgi-biii/npsa jiutomat.pl?page=npsa jm.htnu), accessed from the ExPasy molecular biology server located at the World Wide Web (.expasy.ch/tools/). The analysis indicates that 213P1F11 variant 1 is composed 47.93% alpha helix, 11.57% extended strand, and 40.50% random coil (Figure 12A), variant 2 is composed of 38.70% alpha helix, 9,57% extended strand, and 51.74% random coil (Figure 12B), variant 3 is composed of 50.68% alpha helix, 6.85% extended strand, and 42.47% random coil (Figure 12C), and variant 4 is composed of 39.25% alpha helix, 12. 15% extended strand, and 48.60% random coil (Figure 12D).

Analysis for the potential presence of transmembrane domains in 213P1F1 1 variant 1 was carried out using a variety of transmembrane prediction algorithms accessed from the ExPasy molecular biology server located at the World Wide Web (.expasy.ch/tools/) (Table XXII). The programs do not predict (lie presence of transmembrane domains in any of the 213P1F11 variants, suggesting that each is a soluble protein.

Example 10: Generation of 213P1F11 Polyclonal Antibodies 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 and/or adjuvant will be injected in the mammal by multiple subcutaneous or intraperitoneal injections. In addition to immunizing with die full length 213P 1F1 1 protein, computer algorithms are employed in design of immunogens that, based on amino acid sequence analysis contain characteristics of being antigenic mid available for recognition by the immune system of the immunized host (see the Example entitled "Antigenicity Profiles and Secondary Stnicture")- Such regions would be predicted to be hydrophilic, flexible, in beta -turn conformations, and be exposed on the surface of the protein (see, e.g., Figure 5A-D, Figure 6 A-D, Figure 7 A-D, Figure 8 A-D, or Figure 9 A-D for amino acid profiles that indicate such regions of 213P 1F1 1 and variants).

For example, 213P1F11 recombinant bacterial fusion proteins or peptides containing hydrophilic, flexible, beta-turn regions of 213P1F11 variant proteins are used as antigens to generate polyclonal antibodies in New Zealand White rabbits. For example, such regions include, but are not limited to, amino acids 1-17, amino acids 25-80, amino acids 88-108, amino acids 131 -147, and 207-242 of 213P1F 1 1 variant 1. 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 inliibitor. In one embodiment, a peptide encoding amino acids 1-17 of 213P1F11 variant 1 is conjugated to KLH and used to inunnnize the rabbit. Alternatively the immunizing agent may include all or portions of the 213PIFU variant proteins, analogs or fusion proteins thereof. For example, the 213P1F1 1 variant 1 amino acid sequence can 81 be fused using recombinant DNA techniques 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. Such fusion proteins are purified from induced bacteria using the appropriate affinity matrix.

In one embodiment, a GST-fusion protein encoding amino acids 1-147, encompassing several predicted antigenic regions, is produced and purified and used as immunogen. 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 213P1F11 in Prokaryotic Systems" and Current Protocols In Molecular Biology, Volume 2, Unit 16, Frederick M. Ausubul et al. eds., 1995; Linsley, P.S., Brady, W., Urnes, M, Grosmaire, L., Damle, N., and Ledbetter, L.(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 213P1FI 1 iii Eukaryotic Systems"), and retain post-translational modifications such as glycosylations found in native protein. In one embodiment, the full length sequence of variant 1, amino acids 1-242, is cloned into the Tag5 mammalian secretion vector. The recombinant protein is purified by metal chelate chromatography from tissue culture supernatants of 293T cells stably expressing the recombinant vector. The purified Tag5 213P1 F11 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 ^ig, typically 100-200 ^ig, 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 ^ig, typically 100-200 ig, of the immunogen in incomplete Freund's adjuvant (IF A). Test bleeds are taken approximately 7-10 days following each immunization and used to monitor the titer of die antiserum by ELISA.

To test reactivity and specificity of immune serum, such as the rabbit serum derived from immunization with a KLH-conjugated peptide encoding amino acids 1-17 of variant 1 , the full-length 213P1FI 1 variant 1 cDNA is cloned into pCDNA 3.1 myc-his expression vector (Invitrogen, see the Example entitled "Production of Recombinant '213P IF 1 1 in Eukaryotic Systems"). After transfection of the constructs into 293T cells, cell lysates are probed with the anti-213PlFl 1 senim and with anti-His antibody (Santa Cruz Biotechnologies, Santa Cruz, CA) to determine specific reactivity to denatured 213P1F11 protein using the Western blot technique. The immune serum is then tested by the Western blot technique against 293T-213P1F1 1 cells. In addition, the immune serum is tested by fluorescence microscopy, flow cytometry and immunoprecipitation against 293T and other recombinant 213Pl Fl l-expressing cells to determine specific recognition of native protein. Western blot, immunoprecipitation, fluorescent microscopy, and flow cytometric techniques using cells that endogenously express 213P1F11 are also carried out to test reactivity and specificity. 82 Anti -serum from rabbits immunized with 213PlFl lvariant 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 GST-213P1F 11 fusion protein encoding amino acids 1-147 is first purified by passage over a column of GST protein covalently coupled to AffiGel matrix (BioRad, Hercules, Calif.). The antiserum is then affinity purified by passage over a column composed of a MBP-fusion protein also encoding amino acids 1-147 covalently coupled to Affigel matrix. The serum is then further purified by protein G affinity chromatography to isolate the IgG fraction. Sera from oilier His-tagged antigens and peptide immunized rabbits as well as fusion partner depleted sera are affinity purified by passage over a column matrix composed of die original protein iminunogen or free peptide.

Example 11: Generation ot 213P1F11 Monoclonal Antibodies (mAbs) In one embodiment, therapeutic mAbs to 213P1F1 1 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 die biological function of the 213P1F11 variants, for example those that would dismpt the interaction with ligands and binding partners. Immunogens for generation of such mAbs include dtose designed to encode or contain the entire 213P1F11 protein variant sequence, regions of the 213P1F11 protein variants predicted to be antigenic from computer analysis of the amino acid sequence (see, e.g., Figure 5A-D, Figure 6 A-D, Figure 7 A-D, Figure 8 A-D, or Figure 9 A-D, and the Example entitled "Antigenicity 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 213P1F11 variant, such as 293T-213P1F11 variant 1 or 300.19-213P1F1 1 variant lmurine Pre-B cells, are used to immunize mice.

To generate mAbs to a 213P1F I 1 variant, mice are first immunized intraperitoneal ly (IP) with, typically, 10-50 ^ig of protein iminunogen or It)7 213P1F11 -expressing cells mixed in complete Freund's adjuvant. Mice are then subsequently immunized IP every 2-4 weeks with, typically, 10-50 ng of protein iminunogen 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 wliich a mammalian expression vector encoding a 213P1F11 variant sequence is used to immunize mice by direct injection of the plasmid DNA. For example, the full length variant 1 sequence, encoding amino acids 1-242, is cloned into die Tag5 mammalian secretion vector and die recombinant vector is used as iminunogen. In another example the same amino acids are cloned into an Fc-fusion secretion vector in which the 213P1F11 variant 1 sequence is fused at the amino-terminus to an IgK leader sequence and at die carboxyl-termiuus to die 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 die same vector and wiUi cells expressing die respective 213PlFl lvariant.

During die 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 blotting, immunoprecipilation, fluorescence microscopy, and flow cytometric 83 analyses, fusion and hybridonia generation is then carried out with established procedures well known in the art (see, e.g., Harlow and Lane, 1988).

In one embodiment for generating 213P1F11 monoclonal antibodies, a Tag5-213P 1F11 variant 1 antigen encoding amino acids 1-242, is expressed and purified from stably transfected 293T cells. Balb C mice are initially immunized intraperitoneally with 25 ^ig of die Tag5-213P1F1 1 variant 1 protein mixed in complete Freund's adjuvant. Mice are subsequently immunized every two weeks with 25 ^ig of the antigen mixed in incomplete Freund's adjuvant for a total of three immunizations. ELISA using the Tag5 antigen determines the titer of serum from immunized mice. Reactivity and specificity of serum to full length 213P 1F11 variant protein is monitored by Western blotting, imniunoprecipitation and flow cytometry using 293T cells transfected with an expression vector encoding the 213P1F11 variant 1 cDNA (see e.g., the Example entitled "Production of Recombinant 213P1F11 in Eukaryotic Systems"). Other recombinant 213PlFl lvariant 1-expressiiig cells or cells endogenously expressing 213P1F11 variant 1 are also used. Mice showing the strongest reactivity are rested and given a final injection of Tag5 antigen in PBS and then sacrificed four days later. The spleens of the sacrificed mice are harvested mid fused to SPO/2 myeloma cells using standard procedures (Harlow and Lane, 1988). Supernatants from HAT selected growth wells are screened by ELISA, Western blot, immunoprecipilation, fluorescent microscopy, and flow cytometry to identify 213P1F 11 specific antibody-producing clones.

Monoclonal antibodies are also derived that react only with specific 213P1F11 variants. To tins end, immunogens are designed to encode amino acid regions specific to the respective variant. For example, a Tag5 immunogen encoding amino acids 175-230 of variant 2 is produced, purified, mid used to immunize mice to generate hybridomas. In another example, a KLH-coupled peptide encoding amino acids 135- 146 of variant 3 is produced and used as immunogen. in another example amino acids 1-86 of variant 4 is fused to GST and used as immunogen. Monoclonal antibodies raised to these immunogens are Uien screened for reactivity to cells expressing the respective variants but not to oilier 213P 1F1 1 variants. These strategies for raising 213P1F11 variant specific monoclonal antibodies are also applied to polyclonal reagents described in die Example entitled "Generation of 213P 1F11 Polyclonal Antibodies." The binding affinity of a 213P1F11 monoclonal antibody is determined using standard technologies. Affinity measurements quantify the strength of antibody to epitope binding and are used to help define which 213P1F1 1 monoclonal 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 BIAcore system uses surface plasmon resonance (SPR, Welford K. 1991 , Opt. Quant. Elect. 23 : 1 ; Morton and Myszka, 1998, Metliods in Enzymology 295: 268) to monitor bioinolecular interactions in real time. BIAcore analysis conveniently generates association rate constants, dissociation rate constants, equilibrium dissociation constants, and affinity constants.

Example 12: HLA Class I and Class Π Biiulint; Assays HLA class 1 and class II binding assays using purified HLA molecules are performed in accordance with disclosed protocols (e.g. , PCT publications WO 94/20127 and WO 94/03205; Sidney et ai , Current Protocols in Immunology 18.3.1 (1998); Sidney, et a/., J. Immunol. 154:247 (1995); Sette, et ai, Mol.

Immunol. 31 :813 (1994)). Briefly, purified MHC molecules (5 to 500 nM) are incubated with various 84 unlabeled peptide iiihibitors and 1-10 nM 125l-radiolabeled probe 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 (lie concentration of HLA molecules necessary to bind 10-20% of the total radioactivity. All subsequent inhibition and direct binding assays are performed using these HLA concentrations.

Since under these conditions [label]<[HLA] and IC5o>|HLA], tire measured IC50 values are reasonable approximations of the true KD values. Peptide inliibitors are typically tested at concentrations ranging from 120 to 1.2 lig/nil, mid are tested in two to four completely independent experiments. To allow comparison of tlve data obtained in different experiments, a relative binding figure is calculated for each peptide by dividing the IC5o of a positive control for inliibition by the I.C50 for each tested peptide (typically unlabeled versions of the radiolabeled probe peptide). For database purposes, and inter-experiment comparisons, relative binding values are compiled. These values can subsequently be converted back into IC50 nM values by dividing the IC50 nM of the positive controls for inliibition by the relative binding of the peptide of interest. This method of data compilation is accurate mid 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 supennotif and/or HLA motif-bearing peptides.

Example 13: Identification of HLA Supei motif- and Mutif-Beiiriim 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 die identification and confirmation of supennotif- mid motif-bearing epitopes for die inclusion in such a vaccine composition. Calculation of population coverage is performed using die strategy described below.

Computer searches and algoriduus for identification of supennotif and/or motif-bearing epitopes The searches performed to identify the motif-bearing peptide sequences in1 the Example entitled "Antigenicity Profiles and Secondary Stnicture" mid Tables V-XIX employ the protein sequence data from die gene product of 213P1F11 set forth in Figures 2 mid 3.

Computer searches for epitopes bearing HLA Class I or Class II supennotifs or motifs are performed as follows. All translated 213P1F11 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 accordmice widi information in die ml in view of known motif/supermotif disclosures. Furthermore, such calculations can be made mentally.

Identified A2-, A3-, mid DR-supermotif sequences are scored using polynomial algoriduus to predict dieir capacity to bind to specific HLA-Class I or Class II molecules. These polynomial algoriduus account for the impact of different amino acids at different positions, mid are essentially based on die premise tiiat the overall affinity (or AG) of peptide-HLA molecule interactions can be approximated as a linear polynomial function of the type: "AG" = au x a2, x a3j x a„, 85 where a , is a coefficient which represents the effect of the presence of a given amino acid (J) 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 (i.e., independent binding of individual side-chains). When residue j occurs at position / in the peptide, it is assumed to contribute a constant amount j, to the free energy of binding of the peptide irrespective of the sequence of the rest of the peptide.

The method of derivation of specific algorithm coefficients has been described in Gulukota et ai, J. Mol. Biol. 267: 1258-126, 1997; (see also Sidney et ai, Human Immunol. 45:79-93, 1996; and Soulhwood et al, J. Immunol. 160:3363-3373, 1998). Briefly, for all ; positions, anchor and non-anchor alike, tire geometric mean of the average relative binding (ARB) of all peptides carrying^' is calculated relative to the remainder of the group, and used as the estimate ofy',. For Class II 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 chosen as a function of the degree of stringency of prediction desired.

Selection of HLA-A2 supertype cross-reactive peptides Protein sequences from 213P IF] 1 are scanned utilizing motif identification software, to identify 8-, 9- 10- and 11-mer sequences containing the HLA-A2-supermotif mai anchor specificity. Typically, these sequences are then scored using the protocol described above and tire peptides corresponding to the positive-scoring sequences are synthesized and tested for their capacity to bind purified HLA-A*0201 molecules in vitro (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 mat bind to at least three of the five A2 -supertype alleles tested are typically deemed A2-supertype 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 supermotif-bearinn epitopes The 213P1F11 protein sequence(s) scanned above is also examined for the presence of peptides with die HLA-A3-siipermotif 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* ] 101 molecules, the molecules encoded by the two most prevalent A3-supertype 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 (e.g., 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 epitopes The 213P1FI 1 protein(s) scanned above is also analyzed for the presence of 8-, 9- 10-, or 11-mer peptides with the HLA-B7-supermotif. Corresponding peptides are synthesized and tested for binding to HLA-B*0702, Lire molecule encoded by the most common B7 -supertype allele (i.e. , the prototype B7 supertype allele). Peptides binding B*0702 with iC50 of <500 nM are identified using standard methods. These peptides are then tested for binding to other common B7-supertype molecules (e.g., B*35()l, B*5101 , 86 B*5301 , and B*5401). Peptides capable of binding to three or more of the five B7-supertype alleles tested are thereby identified.

Selection of Al and A24 motif-bearing epitopes To further increase population coverage, HLA-A1 and -A24 epitopes can also be incorporated into vaccine compositions. An analysis of the 213P1F11 protein can also be performed to identify HLA-A1- and A24-motif-containing sequences.

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

Example 14: Confirmation of Immunogenicitv Cross-reactive candidate CTL A2-supermotif-bearing peptides that are identified as described herein are selected to confirm in vitro inmiunogenicity. Confirmation is performed using tlie following methodology: Target Cell Lines for Cellular Screening: The .221A2.1 cell line, produced by transferring the HLA-A2.1 gene into tlie HLA-A, -B, -C null mutant human B-lymphoblastoid cell line 721.221, is used as tlie peptide-loaded target to measure activity of HLA-A2.1 -restricted CTL. This cell line is grown in RPMI-1640 medium supplemented with antibiotics, sodium pyruvate, nonessential amino acids and 10% (v/v) heat inactivated FCS. Cells that express an. antigen of interest, or transfectants comprising the gene encoding the antigen of interest, can be used as target cells to confirm tlie ability of peptide-specific CTLs to recognize endogenous antigen.

Primary CTL Induction Cultures: Generation of Dendritic Cells (DC): PBMCs are thawed in RPMI with 30 DNAse, washed twice and resuspended in complete medium (RPMI-1640 plus 5% AB human serum, non-essential amino acids, sodium pyruvate, L-glutamine and penicillin/streptomycin). The monocytes are purified by plating 10 x 106 PBMC/well in a 6-well plate. After 2 hours at 37°C, (lie non-adherent cells are removed by gently shaking tl e plates and aspirating tlie supernatants. The wells are washed a total of three times with 3 ml RPMI to remove most of tlie non-adherent and loosely adherent cells. Three ml of complete medium containing 50 ng/ml of GM-CSF and 1,000 U/inl of IL-4 are then added to each well. TNFa is added to tlie DCs on day 6 at 75 ng/ml and tl e 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 witli Dynal immui omagnetic beads (Dynabeads© M-450) and the detacha-bead© reagent. Typically about 200-250xl06 PBMC are processed to obtain 24xl05 CD8+ T-cells (enough for a 48-well plate culture). Briefly, tlie PBMCs are thawed in RPMI witli 30μg/lnl DNAse, washed once witli PBS containing 1% human AB serum and resuspended in PBS/1% AB serum at a concentration of 20xl06cells/ml. The magnetic beads are washed 3 times witli PBS/AB serum, added to tlie cells (140μ1 beads/20xl06 cells) and incubated for 1 hour at 4°C witli continuous mixing. The beads and cells are washed 4x witli PBS/AB serum to remove the nonadherent cells and resuspended at ! OOxl O6 cells/ml (based on tlie original cell number) in PBS/AB serum containing 87 ΙΟΟμΙ/ml detacha-bead© reagent and 30 ^ig/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 40ng/ml of peptide at a cell concentration of l -2xl06/ml in (lie presence of 3 ng ml β2- 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 IxlO5 cells/ml) are co-cultured with 0.25ml of CD8+ T-cells (at 2xl06 cell/ml) in each well of a 48-well plate in tire presence of 10 ng/ml of lL-7. Recombinant human IL-10 is added tire next clay at a final concentration of 10 ng/ml and rhuman IL-2 is added 48 hours later at 10 lU/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 RPM1 and DNAse. The cells are resuspended at 5x106 cells/ml and irradiated at ~420() rads. The PBMCs are plated at 2xl06 in 0.5 ml complete medium per well and incubated for 2 hours at 37°C. The plates are washed twice with RPM1 by tapping the plate gently to remove the nonadherent cells and the adherent cells pulsed with lOpg/nrl of peptide in lite presence of 3 μg/ml β2 microglobulin in 0.25ml RPMI/5%AB per well for 2 hours at 37°C. Peptide solution from each well is aspirated and the wells are washed once with RPMI. Most of the media is aspirated from the induction cultures (CD8+ cells) and brought to 0.5 ml with fresh media. The cells are then transferred to tire 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 IL2 is added the next day and again 2-3 days later at 50IU/ml (Tsai et a!. , Critical Reviews in Immunology 18(l-2):65-75, 1998). Seven days later, tire cultures are assayed for CTL activity in a 51 Cr release assay. In some experiments the cultures are assayed for pepude-specific recognition in die 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.

Measurement of CTL lytic activity by S1 Cr release.

Seven days after tire second restimulation, cytotoxicity is determined in a standard (5 lrr) 51 Cr release assay by assaying individual wells at a single E:T. Peptide-pulsed targets are prepared by incubating the cells with ^g/ml peptide overnight at 37°C.

Adherent target cells are removed from culture flasks with trypsin-EDTA. Target cells are labeled with 200 μθί of 51 Cr sodium cluomate (Dupont, Wilmington, DE) for 1 hour at 37°C. Labeled target cells are resuspended at 106 per ml and diluted 1 : 10 witli 562 cells at a concentration of 3.3xl06/ml (an N -sensitive erytl oblastoma cell line used to reduce non-specific lysis). Target cells (100 μΐ) and effectors (ΙΟΟμΙ) are plated in 96 well round -bottom plates and incubated for 5 hours at 37°C. At that time, 100 μΐ of supernatant are collected from each well and percent lysis is determined accordin to tire formula: [(cpm of (lie test sample- cpm of the spontaneous 51 Cr release sample)/(cpm of the maximal 51 Cr release sample- cpm of the spontaneous 51 Cr 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 tire specific lysis (saniple- 88 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-specific and Endogenous Recognition Immulon 2 plates are coated with mouse anti-human ΓΡΝγ monoclonal antibody (4 0.1M NaHC03, pH8.2) overnight at 4°C. The plates are washed with Ca2+, Mg +-free PBS/0.05% Tween 20 and blocked with PBS/10% FCS for two hours, after which the CTLs (100 μΐ/well) and targets (100 μΐ/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 lxlO6 cells/ml. The plates are incubated for 48 hours at 37°C with 5% C02.

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 μΐ of biotinylated mouse anti-human IFN-gamma 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 H3PO4 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 anti-CD3. Briefly, 5x10" CD8+ cells are added to a T25 flask containing the following: lxlO6 irradiated (4,200 rad) PBMC (autologous or allogeneic) per ml, 2xl05 irradiated (8,000 rad) EBV- transformed cells per ml, and OK.T3 (anti-CD3) at 30ng per ml in RPMI-1640 containing 10% (v/v) human AB serum, non-essential amino acids, sodium pyruvate, 25uM 2-mercaptoethanol, L-glutamine and penicillin/streptomycin. Recombinant human IL2 is added 24 hours later at a final concentration of 200IU/ml and ever)' three days thereafter with fresh media at 50IU/ml. The cells are split if the cell concentration exceeds lxl06/ml and the cultures are assayed between days 13 and 15 at E:T ratios of 30, 10, 3 and 1 : 1 in the 51 Cr release assay or at lxl06/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 5xl04 CD8+ cells are added to a T25 flask containing the following: lxlO6 autologous PBMC per ml which have been peptide-pulsed with 10 μg/ml peptide for two hours at 37°C and irradiated (4,200 rad); 2xl05 irradiated (8,000 rad) EBV-transformed cells per ml RPMI-1640 containing 10%(v/v) human AB serum, non-essential AA, sodium pyruvate, 25mM 2-ME, L-glutamine and gentamicin.

Immunogenicity of A2 supermotif-bearing peptides A2-supermotif cross-reactive binding peptides are tested in the cellular assay for die ability to induce peptide-specific CTL in normal individuals. In tliis analysis, a peptide is typically considered to be an epitope 89 if it induces peptide-specific CTLs in at least individuals, and preferably, also recognizes tlie endogenously expressed peptide.

Immunogenicity can also be confirmed using PBMCs isolated from patients bearing a tumor that expresses 213P1F1 1 . Briefly, PBMCs are isolated from patients, re-stimulated with peptide-pulsed monocytes and assayed for tlie ability to recognize peptide-pulsed target cells as well as transfected cells endogenously expressing the antigen.

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

Evaluation of B7 iminiinogenicity Immunogenicity screening of tlie B7-supertype cross-reactive binding peptides identified as set forth herein are confirmed in a maimer analogous to tlie confirmation of A2-and A3 -supennotif-bearing peptides.

Peptides bearing other supermotifs/inotifs, e.g. , HLA-A1, HLA-A24 etc. are also confirmed using similar methodology Example 15: Implementation of the Extended Supermotif to Improve the Binding Capacity of Native Epitopes by Creating Analogs HLA motifs and supermotifs (comprising primary and/or secondary residues) are useful in tlie identification and preparation of highly cross-reactive native peptides, as demonstrated herein. Moreover, tlie 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 tlie 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. ^ Analoging at Primary Anchor Residues Peptide engineering strategies are implemented to furdier increase tlie cross-reactivity of the epitopes. For example, tlie main anchors of A2 -supennotif-bearing peptides are altered, for example,; to introduce a preferred L, I, V, or M at position 2, and I or V at tlie C-terminus.

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

Alternatively, peptide is confirmed as binding one or all supertype members and then analoged to modulate 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 capacity of die parent wild type ( WT) peptide to bind at least weakly, i.e., bind at an IC50 of 5000nM or less, to Uiree of more A2 supertype alleles. The rationale for this requirement is that die WT peptides must be present endogenously in sufficient quantity to be biologically relevant. Analoged peptides 90 have been shown to have increased imnuuiogenicity and cross-reactivity by T cells specific for the parent epitope {see, e.g., Parkhurst et al., J. Immunol. 157:2539, 1996; and Pogue et al, Pro Natl. Acad. Sci. USA 92:8166, 1995).

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

Analoging of HLA-A3 and B7-supermotif-bearing peptides Analogs of HLA-A3 supermotif:bearing epitopes are generated using strategies similar to diose employed in analoging HLA-A2 supermotif-bearing peptides. For example, peptides binding to 3/5 of the A3-superlype molecules are engineered at primary anchor residues to possess a preferred residue (V, 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 siipertype 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 B7-supertype alleles can be improved, where possible, to acliieve increased cross-reactive binding or greater binding affinity or binding half life. B7 supennotif-bearing peptides are, for example, engineered to possess a preferred residue (V, 1, 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 maimer.

The analog peptides are then be confirmed for iimnunogenicity, 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.

Analoging at Secondary Anchor Residues Moreover, HLA supermotifs are of value in engineering higlily 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 peptide is then anaioged to, for example, substitute L for F at position 1 . The anaioged peptide is evaluated for increased binding affinity, binding half life and/or increased cross-reactivity. Such a procedure identifies anaioged peptides with enhanced properties.

Engineered analogs with sufficiently improved binding capacity or cross-reactivity can also be tested for imnuinogenicity in HLA-B7-transgenic mice, following for example, IFA immunization or lipopeptide immunization. Anaioged peptides are additionally tested for the ability to stimulate a recall response using PBMC from patients with 213PlFl l -expressing tumors.

Other analoging strategies 91 Another form of peptide arialogiiig, unrelated to anchor positions, involves the substitution of a cysteine with ot-amino butyric acid. Due to its chemical nature, cysteine has tlie propensity to form disulfide bridges and sufficiently alter tlie 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, e.g. , the review by Sette et ai, In: Persistent Viral Infections, Eds. R. Ahmed and I. Chen, John Wiley & Sons, England, 1999).

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

Example 16: Identification and confirmation of 213PlFl l-tlerived senuenccs with HLA-DR hindinu motifs Peptide epitopes bearing an HLA class II supermotif or motif are identified and confirmed as outlined below using methodology similar to that described for HLA Class I peptides.

Selection of HLA-DR-supermotif-beariiiK epitopes.

To identify 213P IF 1 1 -derived, HLA class II HTL epitopes, a 213P1F11 antigen is analyzed for the presence of sequences bearing an HLA-DR-motif or supermotif. Specifically, 15-mer sequences are selected comprising a DR-supermotif, comprising a 9-mer core, and three-residue N- mid C -terminal flanking regions ( 15 amino acids total).

Protocols for predicting peptide binding to DR molecules have been developed (Southvvood et al., 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 tlie presence of DR-supermotif primary anchors (i.e., 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, e.g., Southwood et ai , ibid. ), it as been found that these protocols efficiently 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 , DR4vv4, and DR7, can efficiently select DR cross-reactive peptides.

The 213P1F11-derived peptides identified above are tested for their binding capacity for various common HLA-DR molecules. All peptides are initially tested for binding to tl e DR molecules in tlie primary 1 panel : DR1, DR4w4, and DR7. Peptides binding at least two of these three DR molecules are Uien tested for binding to DR2w2 β ΐ, DR2w2 β2, DR6wl 9, and DR9 molecules in secondary assays. Finally, peptides binding at least two of tlie four secondary panel DR molecules, and Uius cumulatively at least four of seven different DR molecules, are screened for binding to DR4wl5, DR5vvl 1, and DR8w2 molecules in tertiary assays. Peptides binding at least seven of the ten DR molecules comprising tlie primary, secondary, and tertiary screening assays are considered cross-reactive DR binders. 213P IF 11-derived peptides found to bind common HLA-DR alleles are of particular interest.

Selection of DR3 motif peptides Because HLA-DR3 is an allele dial is prevalent in Caucasian, Black, and Hispanic populations, DR3 binding capacity is a relevant criterion in tlie selection of HTL epitopes. Thus, peptides shown to be candidates may also be assayed for dieir DR3 binding capacity. However, in view of the binding specificity 92 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 213P1F11 antigens are analyzed for sequences carrying one of the two DR3-specific binding motifs reported by Geluk et al. (J. Immunol. 152:5742-5748, 1 94). The corresponding peptides are then synthesized and confirmed as having the ability to bind DR3 with an affinity of Ι μΜ or better, i.e., less than I μΜ. 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 supennotif-bearing peptide epitopes.

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

Example 17: Immunogenicity of 213PlFll-derivecl HTL epitopes This example determines immunogenic DR supermotif- and DR3 motif-bearing epitopes among those identified using the methodology, set forth herein.

Immunogenicity of HTL epitopes are confirmed in a manner analogous to the determination of immunogenicity of CTL epitopes, by assessing the ability to stimulate HTL responses and/or by using appropriate transgenic mouse models. Immunogenicity is determined by screening for: 1.) in vitro primary induction using normal PBMC or 2.) recall responses from patients who have 213P1F11 -expressing tumors.

. Example 18: Calculation of phenotypic frequencies . of HLA-siipcrtypes in various ethnic backgrounds to determine breadth of population coverage This example illustrates the assessment of the breadth of population coverage of a vaccine composition comprised of multiple epitopes comprising multiple supermolifs 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=l-(SQRT(l-af)) (see, e.g., Sidney et al. , Human Immunol. 45:79-93, 1 96). To obtain overall phenotypic frequencies, cumulative gene frequencies are calculated, and the cumulative antigen frequencies derived by the use of the inverse formula [af=l -(l -Cgf)2].

Where frequency data is not available at the level of DNA typing, correspondence to the serologically defined aiitigen 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-loci 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 (e.g., total=A+B*(I-A)). Confirmed members of the A3-like supertype are A3, Al l , A3 1, 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. Likewise, confirmed members of the A2-like supertype family are A*0201, A*0202, A*0203, A*0204, A*0205, A*()206, A*0207, 93 A*6802, and A*6901. Finally, die B7-like supertype-confinned alleles are: B7, B*3501-03, B51, B*5301, B*5401, B*5501 -2, B*5601, B*6701, and B*7801 (potentially also B*1401, B*3504-06, B*4201, and B*5602).

Population coverage achieved by combining the A2-, A3- and B7-superiypes 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 etluiic 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 >95%. An analogous approach can be used to estimate population coverage achieved with combinations of class II motif-bearing epitopes.■ Immunogenicity studies in humans' (e.g., Bertoni el al. , J. Clin. Invest. 100:503, 1997; Doolan et al., Immunity 7:97, 1997; and Tlirelkeld 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 mid from the art), mi average population coverage is predicted to be greater than 95% in each of five major etluiic populations. The game theory Monte Carlo simulation analysis, which is known in the art (see e.g., Osborne, M. J. and Rubinstein, A. "A course in game theory" MIT Press, 1994), can be used to estimate what percentage of the individuals in a population comprised of the Caucasian, North American Black, Japanese, Chinese, and Hispanic etluiic groups would recognize the vaccine epitopes described herein. A preferred percentage is 90%. A more preferred percentage is 95%.

Example 19: CTL Recognition Of Entlogenously Processed Antigens Alter Priming This example confirms that CTL induced by native or analoged peptide epitopes identified and selected as described herein recognize endogenously synthesized, i. e., native antigens.

Effector cells isolated from transgenic mice that are immunized with peptide epitopes, for example HLA-A2 supermotif-bearing epitopes, are re-stimulated in vitro using peptide-coated stimulator cells. Six days later, effector cells are assayed for cytotoxicity and the cell lines that contain peptide-specific cytotoxic activity are fiirdier re-stimulated. An additional six days later, these cell lines are tested for cytotoxic activity on 51 Cr labeled Jurkat-A2.1 Kb target cells in the absence or presence of peptide, and also tested on 51 Cr labeled larget cells bearing the endogenously synthesized antigen, i.e. cells that are stably transfected with 213P 1F11 expression vectors.

The results demonstrate that CTL lines obtained from animals primed with peptide epitope recognize endogenously synthesized 213P1F1 1 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/Kb transgenic mice, several other transgenic mouse models including mice with human Al 1, which may also be used to evaluate A3 epitopes, and B7 alleles have been characterized and others (e.g. , transgenic mice for HLA-A1 94 and A24) are being developed. HLA-DR 1 and HLA-DR3 mouse models have also been developed, which may be used to evaluate HTL epitopes.

Example 20: Activity Of CTL-HTL Conjugated Epitopes In Transgenic M ice This example illustrates the induction of CTLs and HTLs in transgenic mice, by use of a 213P1F11 -derived CTL and HTL peptide vaccine compositions. The vaccine composition used herein comprise peptides to be administered to a patient with a 213P1F11-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 thai enhanced immunogenicity can be acliieved 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 lipidaled, 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 A2. 1 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 lipidated CTL/HTL conjugate, in DMSO/saline, or if tire 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 LPS-aclivated lymphoblasts coated with peptide.

Cell lines: Target cells for peptide-specific cytotoxicity assays are Jurkat cells transfected with the HLA-A2.1/Kb chimeric gene {e.g., Vitiello et al, J. Exp. Med. 173 : 1007, 1991) In vitro CTL activation: One week after priming, spleen cells (30x10s cells/flask) are co-cultured at 37°C with syngeneic, irradiated (3000 rads), peptide coated lymphoblasts (lOxlO6 cells/flask) in 10 ml of culture medium/T25 flask. After six days, effector cells are harvested and assayed for cytotoxic activity. .

. Assay for cytotoxic activity: Target cells (1.0 to 1.5x10s) are incubated at 37°C in the presence of 200 μΐ of 51 Cr. After 60 minutes, cells are washed three times and resuspended in R10 medium. Peptide is added where required at a concentration of 1 ng/ml. For the assay, 10'' 51 Cr-labeled target cells are added to different concenliations of effector cells (final volume of 200 μΐ) in U-bottom 96-well plates. After a six hour incubation period at 37°C, a 0.1 ml aliquot of supernatant is removed from each well mid 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, ,% 51 Cr release data is expressed as lytic units/106 cells. One lytic unit is arbitrarily defined as the number of effector cells required to achieve 30% lysis of 10,000 target cells in a six hour 51 Cr release assay. To obtain specific lytic units/106, tire lytic units/106 obtained in the absence of peptide is subtracted from tire lytic units/106 obtained in the presence of peptide. For example, if 30% 51 Cr release is obtained at the effector (E): target (T) ratio of 50: 1 (i.e., 5xl05 effector cells for 10,000 targets) in die absence of peptide and 5: 1 (i.e., 5xl04 effector cells for 10,000 targets) in the presence of peptide, the specific lytic units would be: [( l/50,000)-( l/500,000)] x 106 = 18 LU. 95 The results are analyzed to assess the magnitude of the CTL responses of animals injected with the immunogenic CTL/HTL 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, mid concomitantly that an HTL response is induced upon administration of such compositions.

Example 21 : Selection of CTL and HTL epitopes for inclusion in a 213PlFl l-spccifk 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 (i.e. , 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 Die selection.

Epitopes are selected which, upon administration, mimic immune responses that are correlated with 213P1F11 clearance. The number of epitopes used depends on observations of patients who spontaneously clear 213P1 F11. For example, if it has been observed that patients who spontaneously clear 213P1F1 1 -expressing cells generate an immune response to at least three (3) from 213P 1F1 1 antigen, then at least three epitopes should be included for HLA class I. A similar rationale is used to determine HLA class II epitopes.

Epitopes are often selected that have a binding affinity of an 1C50 of 500 nM or less for an HLA class 1 molecule, or for class II, an IC50 of 1000 nM or less; or HLA Class I peptides with high binding scores from the B1MAS web site, at URL biinas.dcrt.nih.gov/.

In order to achieve broad coverage of die 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 at least 80% population coverage. A Monte Carlo analysis, a statistical evaluation known in die art, can be employed to assess breadth, or redundancy, of population coverage.

When creating polyepitopic compositions, or a minigene that encodes same, it is typically desirable to generate die smallest peptide possible that encompasses the epitopes of interest. The principles employed are similar, if not the same, as those employed when selecting a peptide comprising nested epitopes. For example, a protein sequence for die vaccine composition is selected because it has maximal number of epitopes contained wiUiin die sequence, i.e. , it has a high concentration of epitopes. Epitopes may be nested or overlapping (i.e., frame slufted 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 mulli-epitopic, peptide can be generated synthetically, recombinanlly, or via cleavage from die native source. Alternatively, an analog can be made of tliis native sequence, whereby one or more of the epitopes comprise substitutions diat alter the cross-reactivity and/or binding affinity properties of the polyepitopic peptide. Such a vaccine composition is 96 administered for therapeutic or prophylactic purposes. This embodiment 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 motif-bearing epitopes for an HLA makeup that is presently unknown. Furthermore, this embodiment (absent tire creating of any analogs) directs tire immune response to multiple peptide sequences that are actually present in 213P1F11, thus avoiding the need to evaluate any junctional epitopes. Lastly, the embodiment provides an economy of scale when producing nucleic acid 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 dial controls or clears cells that bear or overexpress 213P1F11.

Example 22: Construction of "Minigene" Multi-Epitope DNA Plasmids This example discusses the construction of a minigene expression plasrnid. 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 plasrnid typically includes multiple CTL and HTL peptide epitopes. In the present example, HLA-A2, -A3, -B7 supermotif-bearing peptide epitopes and HLA-Al and -A24 motif-bearing peptide epitopes are used in conjunction with DR supermotif-bearing epitopes and/or DR3 epitopes. HLA class I supennotif or motif-bearing peptide epitopes derived.213P lFl l, are selected such that multiple supermotifs/motifs are represented to ensure broad population coverage. Similarly, HLA class II epitopes are selected from 213P1F11 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 die endoplasmic reticulum. For example, the Ii protein may be fused to one or more HTL epitopes as described in the art, wherein the CLIP sequence of the Ii protein is removed and replaced with mi HLA class II epitope sequence so that HLA class II epitope is directed to the endoplasmic reticulum, where the epitope binds to an HLA class II molecules.

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

The minigene DNA plasrnid 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 overlaps, are synthesized and HPLC-purified. The oligonucleotides encode tire selected peptide 97 epitopes as well as appropriate linker nucleotides, Kozak sequence, and signal sequence. The final multiepilope minigene is assembled by extending the overlapping oligonucleotides in three sets of reactions using PCR. A Perkiii/Elmer 9600 PCR machine is used and a total of 30 cycles are performed using the following conditions: 95°C for 15 sec, annealing temperature (5° below the lowest calculated Tin of each primer pair) for 30 sec, and 72°C for 1 mm.

For example, a minigene is prepared as follows. For a first PCR reaction, 5 ^ig of each of two oligonucleotides are annealed and extended: In an example using eight oligonucleotides, i.e., four pairs of primers, oligonucleotides 1+2, 3+4, 5+6, and 7+8 are combined in 100 μΐ reactions containing Pfu polymerase buffer (lx= 10 mM KCL, 10 niM (NH4)2S04> 20 mM Tris-chloride, pH 8.75, 2 niM MgS04, 0.1% Triton X-100, 100 ng/ml BSA), 0.25 mM each dNTP, and 2.5 U of Pfu polymerase. The full-length dimer products are gel-purified, and two reactions containing the product of 1+2 and 3+4, and the product of 5+6 and 7+8 are mixed, annealed, and extended for 10 cycles. Half of the two reactions are then mixed, and 5 cycles of annealing aiid extension carried out before flanking primers are added to amplify the full length product. The full-length product is gel-purified and cloned into pCR-blunt (Invitiogen) and individual clones are screened by sequencing.

Example 23: The Plasmid Construct and the Decree to Which It Induces Immiinoi»enicitv.

The degree to which a plasmid construct, for example a plasmid constructed in accordance with the previous Example, is able to induce immimogenicity is confirmed in vitro by determining epitope presentation by APC following transduction or transfection of the APC with an epitope-expressing nucleic acid constmct. Such a study determines "antigenicity" and allows the use of human APC. The assay determines the ability of the epitope to be presented by the APC in a context thai is recognized by a T cell by quantifying the density of epitope-HLA class I complexes on tire cell surface. Quantitation can be performed by directly measuring the amount of peptide eliited from tire APC {see, e.g. , Sijts 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 tire 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, e.g. , ageyama et ai , J. Immunol. 154:567-576, 1995).

Alternatively, immunogenicily 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 e.g., 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 superinotif peptide to induce CTLs in vivo, HLA-A2.1 Kb transgenic mice, for example, are immunized intramuscularly with 100 ^ig 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 minigene.

Splenocytes from immunized animals are stimulated twice with each of the respective compositions (peptide epitopes encoded in die minigene or the polyepitopic peptide), then assayed for peptide-specific cytotoxic activity in a 51Cr release assay. The results indicate the magnitude of the CTL response directed 98 against tlie A2 -restricted epitope, thus indicating tlie in vivo invmunogenicity of the minigene vaccine and polyepitopic vaccine.

It is, therefore, found that tlie niinigene elicits immune responses directed toward tlie HLA-A2 supermotif peptide epitopes as does tlie polyepitopic peptide vaccine. A similar analysis is also performed using oilier 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 tlie nunigene elicits appropriate immune responses directed toward tlie provided epitopes.

To confirm tlie capacity of a class 11 epitope-encoding mimgene to induce HTLs in vivo, DR transgenic mice, or for those epitopes that cross react with tlie appropriate mouse MHC molecule, I-Ab-restricted mice, for example, are immunized intramuscularly with 100 of plasmid DNA. As a means of comparing tlie 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. CD4+ T cells, i.e. HTLs, are purified from splenocytes of immunized animals and stimulated with each of tlie respective compositions (peptides encoded in tlie nunigene). The HTL response is measured using a 3H-thymidine incorporation proliferation assay, (see, e.g., Alexander et al. Immunity 1 :751-761, 1994). The results indicate tlie magnitude of the HTL response, thus demonstrating tlie in vivo inimunogenicity of the niinigene.

DNA minigenes, constructed as described in tlie previous Example, can also be confirmed as a vaccine in combination with a boosting agent using a prime boost protocol. The boosting agent can consist of recombinant protein (e.g., Barnett e/ al , Aids Res. and Human Retroviruses 14, Supplement 3 :S299-S309, 1998) or recombinant vaccinia, for example, expressing a mimgene or DNA encoding tlie complete protein of interest (see, e.g. , Hanke et al. , Vaccine 16:439-445, 1998; Sedegah et al, Proc. Natl Acad. Sci USA 95 :7648-53, 1998; Hanke and cMichael, Immunol. Letters 66: 177-181, 1999; and Robinson et al , Nature Med. 5:526-34, 1999).

For example, tlie efficacy of the DNA niinigene used in a prime boost protocol is initially evaluated in transgenic mice. In this example, A2.1/Kb transgenic mice are immunized IM with 100 ig of a DNA mimgene encoding tlie immunogenic peptides including at least one HLA-A2 supermotif-bearing peptide. After an incubation period (ranging from 3-9 weeks), tlie mice are boosted IP with 107 pfu mouse of a recombinant vaccinia virus expressing the same sequence encoded by the DNA niinigene. Control mice are immunized with 100 ^ig of DNA or recombinant vaccinia without tlie mimgene sequence, or with DNA encoding tlie niinigene, but without tl e vaccinia boost. After an additional incubation period of two weeks, splenocytes from tl e mice are immediately assayed for peptide-specific activity in an ELISPOT assay.

Additionally, splenocytes are stimulated in vitro with tlie A2-restricted peptide epitopes encoded in tl e niinigene and recombinant vaccinia, then assayed for peptide-specific activity in an alpha, beta and/or gamma 1FN ELISA.

It is found that the mimgene utilized in a prime-boost protocol elicits greater immune responses toward tlie HLA-A2 supermotif peptides than with DNA alone. Such an analysis can also be performed using HLA-A1 1 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 tlie Example entitled "Induction of CTL Responses Using a Prime Boost Protocol." 99 Example 24: Peptide Compositions for Prophylactic Uses Vaccine compositions of the present invention can be used to prevent 213P1F11 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 213PlFl l-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 μ¾ generally 100-5,000 ^tg, 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 epitope-specific 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 213P IF 11 -associated disease.

Alternatively, a composition typically comprising transfecting agents is used for the administration of a nucleic acid-based vaccine in accordance with methodologies known in the art and disclosed herein.

Example 25: Polyepitonic Vaccine Compositions Derived from Native 213P1F11 Sequences A native 213P1F11 polyprotein sequence is analyzed, preferably using computer algorithms defined for each class I and/or class II supermotif 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 is selected; it can be used to generate a minigene construct. The construct is engineered to express tire peptide, which corresponds to die native protein sequence. The "relatively short" peptide is generally less than 250 amino acids in length, often less than 100 amino acids in length, preferably less than 75 amino acids in 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, i.e., it has a high concentration of epitopes. As noted herein, epitope motifs may be nested or overlapping (i.e., frame shifted relative to one another). For example, with overlapping epitopes, tvyo 9-mer epitopes and one 10-nier epitope can be present in a 10 amino acid peptide. Such a vaccine composition is administered for therapeutic or prophylactic purposes.

The vaccine composition will include, for example, multiple CTL epitopes from 213P1F11 antigen and at least one HTL epitope. This polyepitopic native sequence is administered either as a peptide or as a nucleic acid 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 die production of therapeutic or prophylactic immune response-inducing vaccine compositions. Additionally such an embodiment provides for live possibility of motif-bearing epitopes for an HLA makeup(s) that is presently 100 unknown. Furthermore, this embodiment (excluding an analoged embodiment) directs tlie immune response to multiple peptide sequences that are actually present in native 213P 1F11, thus avoiding tlie need to evaluate any junctional epitopes. Lastly, the embodiment provides an economy of scale when producing peptide or nucleic acid vaccine compositions.

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

Example 26: Polyepitopic Vaccine Compositions From Multiple Anti ens The 213P1F11 peptide epitopes of the present invention are used in conjunction with epitopes from other target tiunor-associated antigens, to create a vaccine composition that is useful for tlie prevention or treatment of cancer that expresses 213P1F11 and such other antigens. For example, a vaccine composition can be provided as a single polypeptide that incorporates multiple epitopes from 213P1F11 as well as tumor-associated antigens that are often expressed with a target cancer associated with 213P1F11 expression, or can be administered as a composition comprising a cocktail of one or more discrete epitopes. Alternatively, tlie vaccine can be administered as a minigene constmct 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 tlie presence of specific antibodies, CTL or HTL directed to 213PIF11. Such an analysis can be performed in a manner described by Ogg et ai, Science 279:2103-2106, 1998. In this Example, peptides in accordance with tl e invention are used as a reagent for diagnostic or prognostic purposes, not as an immunogen.

In tliis example highly sensitive human leukocyte antigen tetrameric complexes ("tetramers") are used for a cross-sectional analysis of, for example, 213P1F11 HLA-A*0201 -specific CTL frequencies from HLA A*0201 -positive individuals at different stages of disease or following immunization comprising a 213P 1F1 1 peptide containing an A*0201 motif. Tetrameric complexes are synthesized as described (Musey et ai, N. Engl. J. Med. 337: 1267, 1997). Briefly, purified HLA heavy chain (A*0201 in this example) mid |32-microglobulin are synthesized by means of a prokaryotic expression system. The heavy chain is modified by deletion of the trmisniembrmie-cytosolic tail mid COOH-terminal addition of a sequence containing a BirA enzymatic biotinylation site. The heavy chain, p2-microglobulin, mid peptide are refolded by dilution. The 45-kD refolded product is isolated by fast protein liquid cliromatograpliy and then biotiuylated by BirA in tlie presence of biotin (Sigma, St. Louis, Missouri), adenosine 5 ' triphosphate and magnesium. Streptavidin-phycoerytlirin conjugate is added in a 1 :4 molar ratio, mid tlie tetrameric product is concentrated to 1 nig/nil. The resulting product is referred to as tetramer-phycoerytlirin.

For tl e analysis of patient blood samples, approximately one million PBMCs are centrifuged at 300g for 5 minutes and resuspended in 50 μΐ of cold phosphate-buffered saline. Tri-color analysts is performed with tlie tetrmner-pliycoerythrin, along with anti-CD8-Tricolor, and anti-CD38. The PBMCs are incubated with tetramer and antibodies on ice for 30 to 60 mill and then washed twice before formaldehyde fixation. Gates are applied to contain >99.98% of control samples. Controls for tlie tetramers include both A*0201-negative individuals mid A*0201 -positive non-diseased donors. The percentage of cells stained with tlie 101 tetramer is then determined by flow cytometry. The results indicate the number of cells in die PBMC sample that contain epitope-restricted CTLs, thereby readily indicating the extent of immune response to the 213P1F11 epitope, and thus the status of exposure to 213P1F11, 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 213P1F11 -associated disease or who have been vaccinated with a 213P1F11 vaccine.

For example, the class 1 restricted CTL response of persons who have been vaccinated may be analyzed. The vaccine may be any 213P1F11 vaccine. PBMC are collected from vaccinated individuals and HLA typed. Appropriate peptide epitopes of die invention that, optimally, bear superinotifs 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 (G1BCO Laboratories), resuspended in RPMI-1640 (GIBCO Laboratories) supplemented with L-glutamine (2mM), penicillin (50U/ml), streptomycin (50 mid Hepes (l OmM) containing 10% heat-inactivated human AB serum (complete RPMI) and plated using lnicroculture formats. A synthetic peptide comprising an epitope of the invention is added at 10 ng/ml to each well and HBV core 128-140 epitope is added at 1 μβ/ιηΐ to each well as a source of T cell help during the first week of stimulation.

In the lnicroculture format, 4 x 10s PBMC are stimulated with peptide in 8 replicate cultures in 96-well round bottom plate in 100 μΐ/well of complete RPMI. On days 3 and 10, 100 μΐ of complete RPMI and 20 U/ml final concentration of rIL-2 are added to each well. On day 7 the cultures are transferred into a 96-well flat-bottom plate and restiinulated with peptide, rIL-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 titan 10% specific 51 Cr release, based on comparison with non-diseased control subjects as previously described (Rehermann, et ai, Nature Med. 2: 1 104,1108, 1996; Rehermann ei a/., J. Clin. Invest. 97: 1655- 1665, 1996; and Rehennann et ai. J. Clin. Invest. 98: 1432- 1440, 1996).

Target cell lines are autologous and allogeneic EBV-transformed B-LCL that are eidier purchased from the American Society for Histocompatibility and Iminunogenetics (ASHI, Boston, MA) or established from the pool of patients as described (Guilhot, et al. J. Virol. (56:2670-2678, 1992).

Cytotoxicity assays are performed in the following manner. Target cells consist of either allogeneic HLA-matched or autologous EBV-transformed B lymphoblastoid cell line that are incubated overnight with the synthetic peptide epitope of the invention at 10 μΜ, and labeled with 100 μθ of 51 Cr (Amersham Corp., Arlington Heights, 1L) for 1 hour after which they are washed four times with HBSS.

Cytolytic activity is determined in a standard 4-h, split well 51 Cr release assay using U-bottomed 96 well plates containing 3,000 targets/well. Stimulated PBMC are tested at effector/target (E/T) ratios of 20-50: 1 on day 14. Percent cytotoxicity is determined from the formula: 100 x [(experimental release- 102 spontaneous release)/maximum release-spontaneous release)]. Maximum release is determined by lysis of targets by detergent (2% 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 213P1F11 or a 213PlFU 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 1.5xl05 cells/well and are stimulated with 10 ng/ml syntlietic peptide of the invention, whole 213P1F11 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 lOU/ml IL-2. Two days later, 1 μθ 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 3H-thymidine incorporation. Antigen-specific T cell proliferation is calculated as the ratio of 3H-lhymidine incorporation in the presence of antigen divided by the 3H-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 I D 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 ^i of peptide composition; Group II: 3 subjects are injected with placebo and 6 subjects are injected with 50 ig peptide composition; Group III: 3 subjects are injected with placebo and 6 subjects are injected with 500 ig 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 immunogenicity. 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 die 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 centrifugauon, 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. 103 Example 30: Phase II Trials In Patients Expressing 213P1F11 Phase II trials are performed to study the effect of administering the CTL-HTL peptide compositio s to patients having cancer that expresses 213P 1F11. The main objectives of the trial are to determine an effective dose and regimen for inducing CTLs in cancer patients that express 213P1 F1 1, 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, e.g., by the reduction and/or shrinking of lesions. Such a study is designed, for example, as follows: The studies are performed in multiple centers. The trial design is an open-label, uncontrolled, dose escalation protocol wherein the peptide composition is administered as a single dose followed six weeks later by a single booster shot 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 vvitliin each group range in age from 21-65 and represent diverse ethnic backgrounds. All of them have a tumor that expresses 213P1F1 1.

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 213PlFl l-associated 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 die efficacy of a DNA vaccine in transgenic mice, such as described above in die Example entided "The Plasmid Construct and die Degree to Which It Induces Immunogenicity," can also be used for die 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 mi adjuvant.

For example, die initial immunization may be performed using an expression vector, such as diat constructed in die Example entitled "Construction of "Minigene" Multi-Epilope DNA Plasmids" in die form of naked nucleic acid administered IM (or SC or ID) in die amounts of 0.5-5 mg at nuilliple sites. The nucleic acid (0. 1 to 1000 ng) can also be administered using a gene gun. Following an incubation period of 3-4 weeks, a booster dose is dien administered. The booster can be recombinant fowlpox virus administered at a dose of 5-107 to 5xl09 pfu. An alternative recombinant vims, such as an MVA, canarypox, adenovirus, or adeno-associated virus, can also be used for die booster, or die 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 die initial vaccine and booster doses of the vaccine. Peripheral blood mononuclear cells are isolated from fresh heparinized blood by FicoH-H paque density gradient centrifugation, aliquoted in freezing media and stored frozen. Samples are assayed for CTL and HTL activity.

Analysis of the results indicates diat a magnitude of response sufficient to achieve a therapeutic or protective immunity against 21 3P 1F1 1 is generated. 104 Example 32: Administration of Vaccine Compositions Using Dendritic Cells (DC) Vaccines comprising peptide epitopes of the invention can be administered using APCs, or "professional" APCs such as DC. In this example, peptide-pulsed DC are administered to a patient to stimulate a CTL response in vivo. In this method, dendritic cells are isolated, expanded, and pulsed with a vaccine comprising peptide CTL and HTL epitopes of tlie invention. The dendritic cells are infused back into tlie 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 tlie 213P1F1 1 protein from which the epitopes in tlie 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 Progenipoietin™ (Monsanto, St. Louis, MO) or GM-CSF/IL-4. After pulsing tlie DC with peptides, and prior to reinfiision into patients, tlie DC are washed to remove unbound peptides.

As appreciated clinically, and readily determined by one of skill based on clinical outcomes, tlie number of DC reinfused into tlie patient can vary (see, e.g., 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 10s can also be provided. Such cell populations typically contain between 50-90% DC.

In some embodiments, peptide-loaded PBMC are injected into patients without purification of tlie . DC. For example, PBMC generated after treatment with an agent such as Progenipoietin™ are injected into · patients without purification of the DC. The total number of PBMC that are administered often ranges from 10B to 1010. Generally, tlie cell doses injected into patients is based on tlie percentage of DC in tlie blood of each patient, as determined, for example, by immunofluorescence analysis with specific anti-DC antibodies. Thus, for example, if Progenipoietin™ mobilizes 2% DC in the peripheral blood of a given patient, and that patient is to receive 5 x 106 DC, then tlie patient will be injected with a total of 2.5 x 108 peptide-loaded PBMC. The percent DC mobilized by an agent such as Progenipoietin™ is typically estimated to be between 2-10%, but can vary as appreciated by one of skill in the art.

Ex vivo activation of CTL/HTL responses Alternatively, ex vivo CTL or HTL responses to 213P1F11 antigens can be induced by incubating, in tissue culture, tlie patient's, or genetically compatible, CTL or HTL precursor cells together with a source of APC, such as DC, and immiuiogenic peptides. After an appropriate incubation time (typically about 7-28 days), in wliich tlie precursor cells are activated and expanded into effector cells, tlie cells are infused into tlie patient, where they will destroy (CTL) or facilitate destruction (HTL) of their specific target cells, i.e., tumor i 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 HLA molecule. These cells can be transfected with nucleic acids that express 105 tlie antigen of interest, e.g. 213P1F11. 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 tlie cell 's surface. Peptides are then eluted from die HLA molecules by exposure to mild acid conditions and their amino acid sequence determined, e.g. , by mass spectral analysis (e.g., Kubo et al, J. Immunol. 152:3913, 1994). Because tlie majority of peptides Uiat bind a particular HLA molecule are motif-bearing, this is an alternative modality for obtaining tlie motif-bearing peptides correlated with tlie particular HLA molecule expressed on tlie cell.

Alternatively, cell lines that do not express endogenous HLA molecules can be tnmsfected with an expression construct encoding a single HLA allele. These cells can then be used as described, i. e., they can then be transfected with nucleic acids that encode 213P1F11' to isolate peptides corresponding to 213P1F11 that have been presented on tlie cell surface. Peptides obtained from such an analysis will bear motif(s) mat correspond to binding to tl e single HLA allele that is expressed in tlie cell.

As appreciated by one in die art, one can perform a similar analysis on a cell bearing more than one HLA allele and subsequently determine peptides specific for each HLA allele expressed. Moreover, one of skill would also recognize diat means other than transfection, such as loading widi a protein antigen, can be used to provide a source Of antigen to die cell.

Example 34: Complementary Polynucleotides Sequences complementary to die 213P IF 11 -encoding sequences, or any parts Uiereof, are used to detect, decrease, or inhibit expression of naturally occurring 213P1F11. Although use of oligonucleotides comprising from about 15 to 30 base pairs is described, essentially tlie same procedure is used with smaller or with larger sequence fragments. Appropriate oligonucleotides are designed using, e.g., OLIGO 4.06 software (National Biosciences) and die coding sequence of 213P1F11. To inhibit transcription, a complementary oligonucleotide is designed from tlie most unique 5' sequence and used to prevent promoter binding to d e coding sequence. To inliibit translation, a complementary oligonucleotide is designed to prevent ribosomal binding to a 213P1F1 1 -encoding transcript.

Example 35: Purification of Naturally-occurring or Recombinant 213P1F11 Using 213P1F11-Specific Antibodies Naturally occurring or recombinant 213P1F11 is substantially purified by inimunoaffmity chromatography' using antibodies specific for 213P1 F11. An invmunoaffinity column is constructed by covalently coupling anti-213PlFl 1 antibody to an activated chromatographic resin, such as CNBr-activated SEPHAROSE (Amersham Pharmacia Biotech). After die coupling, d e resin is blocked and washed according to die manufacturer's instructions.

Media containing 213P1F11 are passed over die inimunoaffmity column, and die column is washed under conditions dial allow the preferential absorbance of 213P1F1 1 (e.g., high ionic strength buffers in die presence of detergent). The column is eluted under conditions dial disrupt antibody/213P 1F1 1 binding (e.g., a buffer of pH 2 to pH 3, or a high concentration of a chaouope, such as urea or diiocyanate ion), and GCR.P is collected. 106 Example 36: Identification of Molecules Which Interact with 213P1F11 213P1F11, or biologically active fragments thereof, are labeled with 121 1 Bolton-Hunter reagent. (See, e.g., Bolton et al. (1973) Biochem. J. 133 :529.) Candidate molecules previously arrayed in tlie wells of a multi-well plate are incubated with tlie labeled 213P1F11, washed, and any wells with labeled 213P 1F11 complex are assayed. Data obtained using different concentrations of 213P 1F11 are used to calculate values for tlie number, affinity, and association of 213P1F11 with tlie candidate molecules.

Example 37: In Vivo Assay for 213P1F11 Tumor Growth Promotion The effect of the 213P1F11 protein on tumor cell growth is evaluated in vivo by ev luating tumor development and growth of cells expressing or lacking 213P1F11. For example, SC1D mice are injected siibcutaneously on each flank with 1 x 106 of either prostate, bladder or breast cancer cell lines (such as PC3, DU145, UM-UC3, J82, MCF7) or NIH-3T3 cells containing tkNeo empty vector or 213P1F1 1. At least two strategies may be used: (1) Constitutive 213P1F11 expression under regulation of a promoter such as a constitutive promoter obtained from tlie genomes of viruses such as polyoma virus, fowlpox virus (UK 2,21 1,504 published 5 July 1 89), adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, liepautis-B virus and Simian Virus 40 (SV40), or from heterologous mammalian promoters, e.g., the actin promoter or an immunoglobulin promoter, provided such promoters are compatible with tlie host cell systems, and (2) Regulated expression under control of an inducible vector system, such as ecdysone, tet, etc., provided such promoters are compatible with tlie host cell systems. Tumor volume is then monitored at tlie appearance of palpable tumors and followed over time to determine if 213P1F11 -expressing cells grow at a faster rate and whedier tumors produced by 213P1F11 -expressing cells demonstrate characteristics of altered aggressiveness (e.g. enhanced melastasis, vascularization, reduced responsiveness to chemotherapeutic drugs).

Additionally, mice can be implanted with 1 x 105 of tlie same cells ortliotopically to determine if 213P1F11 has an effect on local growth in tlie prostate or on tlie ability of the cells to metastasize, specifically to lungs, lymph nodes, and bone marrow.

The assay is also useful to determine tlie 213P1F11 inhibitory effect of candidate therapeutic compositions, such as for example, 213P 1 F I 1 intrabodies, 213P1F11 anlisense molecules and ribozynies.

Example 38: 213P1F11 Monoclonal Antibody-mediated Inhibition of Tumors In Vivo The significant expression of 213P 1F11 in cancer tissues, together with its restricted expression in normal tissues, makes 213P1F11 an excellent target for antibody therapy. In cases where' tlie monoclonal antibody target is a cell surface protein, antibodies have been shown to be efficacious at inhibiting tumor growth (See, e.g., (Saffran, D., et al., PNAS 10: 1073-1078 or tlie World Wide Web at .pnas.org/cgi/doi/10.1073/pnas.051624698). In cases where tlie target is not on tlie cell surface, such as for 213P1F11 mid including PSA and PAP in prostate cancer, antibodies have still been shown to recognize and inhibit growth of cells expressing those proteins (Saffran, D.C., et al., Cancer and Metastasis Reviews, 1999. 18: p. 437-449). As with any cellular protein with a restricted expression profile, 213P1F11 is a target for T cell-based immunotherapy. 107 Accordingly, tlie therapeutic efficacy of anti-213PlFl 1 mAbs in human prostate, bladder and breast cancer mouse models is investigated using in 213P IF 11 -expressing prostate and bladder cancer xenografts as well as prostate, bladder and breast cancer cell lines, such as those described in tlie Example entitled "In Vivo Assay for 213P1F11 Tumor Growth Promotion," that have been engineered to express 213P1F11.

Antibody efficacy on tumor growth and metastasis formation is confirmed, e.g., in a mouse orthotopic prostate or bladder cancer xenograft models, as well as SC1D mice injected with prostate, bladder and breast cancer cell lines, such as those described in tlie Example entitled "In Vivo Assay for 213P1 F11 Tumor Growth Promotion," designed to express or lack 213P1F1 1. Therapeutic efficacy of anti-213PlFl l mAbs in prostate cancer is also evaluated in human prostate xenograft mouse models sucha s tlie LAPC-9 xenografts (Craft, N., et al.,. Cancer Res, 1999. 59(19): p. 5030-6). The antibodies can be unconjugated, as discussed in this Example, or can be conjugated to a therapeutic modality, as appreciated in tlie art. It is confirmed that anti-213PlFl l mAbs inlubit formation of 213P1F11 -expressing tumors. Anti-213P1F11 mAbs inhibit formation of tlie androgen-indepeiident LAPC-9-AI tumor xenografts, as well as PC3-213P1F1 1, MCF7-213P1F11 and UM-UC3-213P1F11 tumors. Aiiti-213P1F11 mAbs also retard tlie growth of established orthotopic tumors and prolong survival of tumor-bearing mice. These results indicate tlie utility of anti-213PlFl 1 mAbs in tlie treatment of local mid advanced stages of prostate, bladder or breast cancer. (See, e.g., Saffran, D., et al., PNAS' 10:1073-1078 or tlie World Wide Web at ,pnas.org/cgi/doi/10.1073/pnas.051624698) Administration of anti-213PlFl l mAbs retard established orthotopic tumor growth and inhibit metastasis to distant sites, resulting in a significant prolongation in tlie survival of tumor-bearing mice. These studies indicate that 213P1F1 1 is an attractive target for immunotherapy and demonstrate tlie therapeutic potential of anti-213PlFl 1 mAbs for tlie treatment of local and metastatic bladder cancer.

This example demonstrates that unconjugated 213P1F11 monoclonal antibodies effectively to inhibit tlie growth of human prostate, bladder and breast tumors grown in SCI D mice; accordingly a combination of such efficacious monoclonal antibodies is also effective.

Tumor inhibition using multiple unconjugated 213P1F11 mAbs Materials and Methods 213P1F11 Monoclonal Antibodies: Monoclonal antibodies are raised against 213P1F11 as described in tlie Example entitled "Generation of 213P1F11 Monoclonal Antibodies (mAbs)." The antibodies are characterized by ELISA, Western blot, FACS, and immunoprecipitation , in accordance with teclmiques known in the art, for then-capacity to bind 213P1F1 1. Epitope mapping data for tlie anti-213PlFl l mAbs, as determined by ELISA and Western analysis, recognize epitopes on tlie 213P1F1 1 protein. Immuiiohistochemical analysis of bladder cancer tissues and cells with these antibodies is performed.

• The monoclonal antibodies are purified from ascites or hybridoma tissue culture siipernatants by Protein-G Sepharose 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 mid used for the treatment of mice receiving subcutaneous or orthotopic injections of bladder tumor xenografts. 108 Cancer Xenograft and Cell Lines The LAPC-9 xenograft, which expresses a wild-type androgen receptor and produces prostate-specific antigen (PSA), is passaged in 6- to 8-week-old male ICR-severe combined inmmnodeficient (SCID) mice (Taconic Farms) by s.c. trocar implant (Craft, N., et al., supra). Prostate, bladder or breast cancer cell lines (such as PC3, DU145, UM-UC3, J82, CF7) expressing 213P1F11 are generated by retroviral gene transfer as described in Hubert, R.S., et al., STEAP: a prostate-specific cell-surface antigen highly expressed in human prostate tumors. Proc Natl Acad Sci U S A, 1999. 96(25):14523-8. Anti-213P1F11 staining is detected by using an FITC-conjugated goat anti-mouse antibody (Southern Biotechnology Associates) followed by analysis on a Coulter Epics-XL f low cytometer.

In Vivo Mouse Models.

Subcutaneous (s.c.) tumors are generated by injection of 1 x 10 6 213P1F11 -expressing cancer 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 die same day as tumor-cell injections. As a control, mice are injected witli either purified mouse IgG (ICN) or PBS; or a purified monoclonal antibody that recognizes an irrelevant antigen not expressed in human cells, hi preliminary studies, no difference is found between mouse IgG or PBS on tumor growth. Tumor sizes are determined by vernier caliper measurements, and tlie tumor volume is calculated as length x width x height. Mice with s.c. tumors greater than 1.5 cm in diameter are sacrificed. Circulating levels of anti-213PlFl 1 mAbs are determined by a capture ELISA kit (Bethyl Laboratories, Montgomery, TX). (See, e.g., (Saffran, D., et al., PNAS 10: 1073-1078) Ortliotopic injections are performed, for example, in two alternative embodiments, under anesthesia by, for example, use of ketamine/xylazine. In a first embodiment, an intravesicular injection of bladder cancer cells is administered directly tlirough tlie urethra and into tlie bladder (Peralta, E. A., et al., J. Urol., 1999. 162: 1806-181 1 ). In a second embodiment, an incision is made tlirough the abdominal wall, tlie bladder is exposed, and bladder tumor tissue pieces (1 -2 mm in size) derived from a s.c. tumor are surgically glued onto die exterior wall of tlie bladder, termed "onplanlation" (Fu, X., et al., Int. J. Cancer, 1991. 49: 938-939; Chang, S., et al., Anticancer Res., 1997. 17: p. 3239-3242). For prostate orthotopic studies, an incision is made through die abdominal muscles to expose the bladder and seminal vesicles, which then are delivered tlirough tlie incision to tlie exposed the dorsal prostate. Antibodies can be administered to groups of mice at tl e time of tumor injection or onplantation, or after 1-2 weeks to allow tumor establishment.

Anti-213P1F1 1 mAbs Inhibit Growth of 213P1F1 1 -Expressing Cancer Tumors In one embodiment, tlie effect of anti-213PlFl l mAbs on tumor formation is tested by using tlie prostate and bladder ortliotopic models. As compared with tlie s.c. tumor model, tlie ortliotopic model, which requires surgical attachment of tumor tissue directly on tlie prostate or bladder, results in a local tumor growth, development of metastasis in distal sites, and subsequent death (Fu, X., et al., Int. J. Cancer, 1991. 49: p. 938-939; Chang, S., et al., Anticancer Res., 1997. 17: p. 3239-3242). This feature make the orUiotopic model more representative of human disease progression and allows one to follow the Uierapeutic effect of mAbs, as well as other tiierapeutic modalities, on clinically relevant end points.

Accordingly, 213P IF 11 -expressing tumor cells are implanted orthotopically, and 2 days later, the mice are segregated into two groups and treated with either: a) 50-2000μg, usually 200-500ng, of anti- 109 213P1F11 Ab, or b) PBS, three times per week for two to five weeks. Mice are monitored weekly for indications of tumor growth. ■ As noted, a major advantage of the ortliotopic prostate and bladder cancer models is the ability to study the development of metastases. Fonnation of metastasis in mice bearing established ortliotopic tumors is studied by histological analysis of tissue sections, including lung and lymph nodes (Fu, X., et ai, Int. J. Cancer, 1991. 49:938-939; Chang, S., et ai, Anticancer Res., 1997. 17:3239-3242). Additionally, IHC analysis using anti-213PlFl 1 antibodies can be performed on the tissue sections.

Mice bearing established orthotopic 213P1F11 -expressing tumors are administered lOOOng injections of either anti-213PIF11 mAb or PBS over a 4-week period. Mice in both groups are allowed to establish a high tumor burden (1-2 weeks growth), to ensure a high frequency of metastasis formation in mouse lungs and lymph nodes. Mice are then sacrificed and their local bladder tumor and lung and lymph node tissue are analyzed for the presence of tumor cells by histology and IHC analysis.

These studies demonstrate a broad anti-tumor efficacy of anti-213PlFl 1 antibodies on initiation and progression of bladder cancer in mouse models. Anti-213P1F11 antibodies inhibit tumor formation and retard the growth of already established tumors and prolong the survival of treated mice. Moreover, anti-213P1F1 1 mAbs demonstrate a dramatic inhibitory effect on the spread of local prostate, bladder and breast tumors to distal sites, even in the presence of a large tumor burden. Thus, anti-213P1F11 mAbs are efficacious on major clinically relevant end points including lessened tumor growth, lessened metastasis, and prolongation of survival.

Example 39: Therapeutic and Diagnostic use of Anti-213P1F11 Antibodies in Humans.

Anti-213P1F11 monoclonal antibodies are safely and effectively used for diagnostic, prophylactic, prognostic and/or therapeutic purposes in humans. Western blot and immunohistochemical analysis of cancer tissues m d cancer xenografts with anti-213PlFl 1 mAb show strong extensive staining in carcinoma but significantly lower or undetectable levels in normal tissues. Detection of 213P1F11 in carcinoma and in metastatic disease demonstrates the usefulness of the mAb as a diagnostic and or prognostic indicator. Anti-213P1 F 1 1 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-213P1F1 1 mAb specifically binds to carcinoma cells. Thus, cUiti-213P1F 1 1 antibodies are used in diagnostic whole body imaging applications, such as radioimmunoscintigraphy and radioiminunotiierapy, (see, e.g., Potamianos S., et. al. Anticancer Res 20(2A):925-948 (2000)) for the detection of localized and metastatic cancers that exhibit expression of 213P1F11. Shedding or release of an extracellular domain of 213P 1F11 into the extracellular milieu, such as that seen for alkaline phosphodiesterase BIO (Meerson, N. R., Hepatology 27:563-568 (1998)), allows diagnostic detection of 213P1F11 by anti-213P lFI l antibodies in serum and/or urine samples from suspect patients.

Anti-213P1F11 antibodies that specifically bind 213P1F11 are used in therapeutic applications for die treatment of cancers that express 213P1F11. Anti-213P1F11 antibodies are used as an unconjugated modality and as conjugated form in wliich the antibodies are atlached to one of various therapeutic or imaging modalities well known in the art, such as a prodrugs, enzymes or radioisotopes. In preclinical studies, unconjugated and conjugated anti-213PlFll antibodies are tested for efficacy of minor prevention and growth inhibition in the SCID mouse cancer xenograft models, e.g., kidney cancer models AGS- 3 and AGS- .6, (see, e.g., the Example entitled "213P1F11 Monoclonal Antibody-mediated Inhibition of Bladder and Lung Tumors In Vivo "). Conjugated and unconjugated anti-213PlFl 1 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-213P1F11 Antibodies In vivo Antibodies are used in accordance with the present invention which recognize an epitope on 213P1F11, and are used in the treatment of certain tumors such as those, listedin Table 1. Based upon a number of factors, including 213P1F11 expression levels, tumors such as those listed in Table I are presently preferred indications. In connection with each of tliese indications, three clinical approaches are successfully pursued.

I. ) Adjunctive therapy: In adjunctive therapy, patients are treated with anli-213PlFl l antibodies in combination with a chemotherapeutic or antineoplastic agent and/or radiation therapy. Primary cancer targets, such as those listed in Table I, are treated under standard protocols by the addition anti- 213P1F11 antibodies to standard first and second line therapy. Protocol designs address effectiveness as assessed by reduction in tumor mass as well as die 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-213P1F1 1 antibodies are utilized in several adjunctive clinical trials in combination with the chemotherapeutic or antineoplastic agents adriamycin (advanced prostrate carcinoma), cisplatin (advanced head and neck and lung carcinomas), taxol (breast cancer), and doxorubicin (preclinical).

II. ) Monotherapy: In connection with tire use of die anti-213PlFl 1 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 radionuclide (e.g., iodine or yttrium (I131, Y90) to anti-213P1F11 antibodies, d e radiolabeled antibodies are utilized as a diagnostic and/or imaging agent. In such a role, die labeled antibodies localize to both solid tumors, as well as, metastatic lesions of cells expressing 213P1F11. In connection with d e use of die aiiti-213P1F11 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 postoperative follow-up to determine what tumor remains and/or returns. In one embodiment, a (i n Iii)-213P 1F11 antibody is used as an imaging agent in a Phase 1 human clinical trial in patients having a carcinoma that expresses 213P1FU (by analogy see, e.g. , 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 I 'll As appreciated by those of ordinary skill in the art, dosing considerations can be determined through comparison with (l e analogous products that are in the clinic. Thus, anti-213PlFl 1 antibodies can be administered with doses in the range of 5 to 400 mg/m 2 , with the lower doses used, e.g., in connection with safety studies. The affinity of anti-213PlFl 1 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-213PlFl l antibodies that are fully human antibodies, as compared to the chimeric antibody, have slower clearance; accordingly, dosing in patients with such fully human anti-213PlFl 1 antibodies can be lower, perhaps in the range of 50 to 300 mg/m2 , and still remain efficacious. Dosing in mg/m2 , 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.

Three distinct delivery approaches are useful for deliver)' of anti-213P'lFl l antibodies.

Conventional intravenous 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 liigh dose of antibody at the tumor and to also minimize antibody clearance. In a similar maimer, certain solid tumors possess vasculature that is appropriate for regional perfusion. Regional perfusion allows for a high dose of antibody at 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-213PlFI 1 antibodies in connection with adjunctive therapy, monotherapy, and as an imaging agent. Trials initially demonstrate safety and thereafter confirm efficacy in repeat doses. Trails are open label comparing standard chemotherapy with standard therapy plus anti-213PlFU antibodies. As will be appreciated, one criteria that can be utilized in connection with enrollment of patients is 213P1F11 expression levels in their tumors as determined by biopsy.

As with any protein or antibody infusion-based therapeutic, safety concerns are related primarily to (i) cytokine release syndrome, i.e., hypotension, fever, shaking, chills; (ii) the development of an immunogenic response to the material (i.e., development of human antibodies by the patient to the antibody therapeutic, or HAHA response); and, (iii) toxicity to normal cells that express 213P1F11. Standard tests and follow-up are utilized to monitor each of these safety concerns. Anti-213P1F11 antibodies are found to be safe upon human administration.

Example 41: Human Clinical Trial Ad junctive Therapy with Human Anti-213P1F11 Antibody and Chemotherapeutic Agent A phase I human clinical trial is initiated to assess the safety of six intravenous doses of a human anti-213PlFl 1 antibody in connection with the treatment of a solid tumor, e.g., a cancer of a tissue listed in Table 1. In the study, the safety of single doses of anti-213PlFl 1 antibodies when utilized as an adjunctive therapy to an antineoplastic or chemotherapeutic agent, such as cisplatin, topotecan, doxorubicin, adriamycin, taxol, or the like, is assessed. The trial design includes delivery of six single doses of an anti-213PlFl 1 antibody with dosage of antibody escalating from approximately about 25 mg/m 2 to about 275 mg/m over die course of the treatment in accordance with the following schedule: 112 Day O Day 7 Day 14 Day 21 Day 28 Day 35 mAb Dose 25 75 125 175 225 275 ' mg/m 2 mg/m 2 mg/m 2 mg/m 2 mg/m 2 mg/m 2 Chemotherapy + + + + + + (standard dose) Patients are closely followed for one-week following each administration of antibody and chemotlierapy. In particular, patients are assessed for the safety concerns mentioned above: (i) cytokine release syndrome, i.e., hypotension, fever, shaking, cliills; (ii) the development of an immunogenic response to the material (i.e., development of human antibodies by the patient to the human antibody therapeutic, or HAHA response); and, (iii) toxicity to normal cells that express 213P1F11. 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 MRJ or other imaging.

The anti-213PlFl 1 antibodies are demonstrated to be safe and efficacious, Phase II trials confirm the efficacy and refine optimum dosing.

Example 42: Human Clinical Trial: Monotherapy with Human Anti-213P1F11 Antibody Anti-213P IF 11 antibodies are safe in connection wit the above-discussed adjunctive trial, a Phase II human clinical trial confirms the efficacy and optimum dosing for monotherapy. Such trial is accomplished, and entails the same safety and outcome analyses, to tire above-described adjunctive trial with die exception being that patients do not receive chemotherapy concurrently with the receipt of doses of anti-213P 1F11 antibodies.

Example 43: Human Clinical Trial: Diagnostic Imaging with Anti-213P1F11 Antibody Once again, as the adjunctive therapy discussed above is safe within the safety criteria discussed above, a human clinical trial is. conducted concerning the use of anti-213P lFl l antibodies as a diagnostic imaging agent. The protocol is designed in a substantially similar manner to those described in the art, such as in Divgi el al. J. Natl. Cancer Inst. 83 :97-104 (1991). The antibodies are found to be both safe and efficacious when used as a diagnostic modality.

Example 44: Homology Comparison of 213P1F11 to Known Sequences The 213P1F11 gene is homologous to a previously cloned gene, namely the human caspase 14 precursor (gi 6912286) (Hu S et al, J. Biol. Chem. 1998, 273 :29648), also known as mini-ICE (MICE). The 213P1F11 gene resulted in several protein variants, which share several characteristics (Table XXll), including homology to ICE family of cysteine proteases. Several variants of 213P1F11,. namely 213P1F11-v.2, -V.3 and -v.4, are novel proteins that maintain some homology to the published caspase 14 precursor (gi 6912286). For example, 213P1F11-V.2 shows 100% identity to the human caspase 14 precursor (gi 6912286) over the first 174 aa of the protein (Figure 4D), while differing from die published caspase 14 113 precursor protein by 56 amino acids at its C-terminus, thus resulting in 76% overall identity to caspase 14 precursor. 213P1F11-V.2 also maintains homology to tire mouse caspase 14, and shows 83% homology and 72% identity to that protein (gi 6753280) (Figure 4F). The 213P1F1 l-v.3 variant protein show 100% identity to the human caspase 14 precursor (gi 6912286) over 134 amino acids, while differing from thai protein by 12 aa at its C-terminus. Similarly, 213P1F11-v.4 shows 97% identity with the human caspase 14 precursor over 235 amino acids, while differing from the human caspase 14 precursor (gi 6912286) by 86 a at its N-terminus (Figure 4G). 213P1F11-v. 1 consists of 242 amino acids, widi calculated molecular weight of 28.0 kDa, and pi of 5.4. 213P1F11-v.1 is an intercellular protein, located in the cytosol with potential localization to tire nucleus (Table XXII). Similar localization patterns are observed for 213P 1F1 1 protein variants 1 , 3, and 4 (Table XXII). Bioinformatic analysis indicates that 213P1 F1-V.2 may also localize to the mitochondria (Table XXII).

Caspases are a family of cyteine proteases that function as effectors of apoptosis or programmed cell death (Salvesen GS, Dixit V, Cell. 1997, 91 :443; Thombeny N, Lazebnik Y, Science. 281 : 1312). These proteases cleave different cellular substrates in an aspartate-specific maimer. Cleavage may result in activation or inactivation of the cleaved cellular proteins, but not in protein degradation (Nunez et al, Oncogene. 1998, 17:3237; Slennicke HR, Salvesen GS, Cell Death Differ 1999, 6: 1054). Caspases traditionally exist as precursor proteins also known as single polypeptide zymogens consisting of a pro-domain, and 2 catalytic subunits, p20 and plO and contain a conserved QACXG active site (Slennicke HR, Salvesen GS, Cell Death Differ 1999, 6: 1054; Cohen M. Biochem J 1997, 326: 1). Similar to other members of the caspase family, 213P1F11 contains two catalytic subunits, p20 and plO, in addition to the conserved penia-peptide active site. In 213P1F11 v.1, p20 spans aa 16-139 and plO spans aa 155-241, while tire active site is located at aa 129. Similarly, 213P1F11 v.4 carries both p20 and plO subunits, while 213P1F1 1 v.2 and v.3 contain the p20 subiuiit only, indicating that all 4 variants of 213P1F11 can function in a similar maimer. Caspases are activated by proteolytic cleavage of their internal aspartate by an upstream enzyme, often another caspase. However, unlike oilier caspases with short pro-domains, caspase 14 is not reported to associate with known caspases (Hu S et al, J. Biol. Chem. 1998, 273 :29648). Caspase 14 has been shown to be processed by caspase 8 and caspase 10 as well as granzyme B, resulting in two catalytic subunits, p20 mid plO (Ahmad M et al, Cancer Res. 1998, 58:5201). These 2 cleavage products are detected in human epidermis and in vitro during keratinocyte differentiation (Eckhart L et al, J. Invest. Dnnatol. 2000, 1 15: 1148). Overexpression of caspase 14 in breast carcinoma cells MCF7 resulted in die apoptosis of these cells, suggesting that caspase 14 participates in (lie process of programmed cell deadi (Hu S et al, J. Biol. Chem. 1998, 273 :29648).

Our findings that 213P1F11 is highly expressed in several cancers while showing a restricted expression pattern in normal tissues suggests that tire 213P1F11 gene plays an important role in various cancers, including cancers of the prostate, bladder and breast. Based on its similarity to caspase 14 213P1F11 has the ability to control tumor growth, apoptosis, survival, differentiation and progression. Accordingly, when 213P1F1 1 functions as a regulator of cell growth and apoptosis, or expression, 213P1F11 is used for therapeutic, diagnostic, prognostic or preventative purposes. 114 Our findings that 213P1F11 is highly expressed in several cancers while showing a restricted expression pattern in normal tissues suggests that the 2I3P1F11 gene plays an important role in various cancers, including cancers of the prostate, bladder mid breast. Based on its similarity to caspase 14 213P 1F1 1 has die ability to control tumor growth, apoptosis, survival, differentiation and progression. Accordingly, when 213P1F11 functions as a regulator of cell growth and apoptosis, or expression, 213P1F11 is used for therapeutic, diagnostic, prognostic or preventative purposes. _ , Example.45: Identification and Confirmation of Signal Transduction Pathways Many mammalian proteins have been reported to interact with signaling molecules and to participate in regulating signaling pathways. (J Neurochem. 2001; 76:217-223). Caspases participate in signal transduction processes by getting recruited to signaling complexes and cleaving specific cellular substrates including other caspases and structural proteins, ultimately resulting in morphologic changes that represent the hallmark of apoptosis (Cohen GM. Biochem J. 1997, 326: 1). Recent studies have demonstrated that caspases also cleave signaling molecules, such as the guanine nucleotide exchange factor TIAMl, leading to the inactivatioii of TIAMl and thereby the Rac cascade (Qi H et al, Cell Growth Differ. 2001, 12:603). Using immunoprecipitation and Western blotting techniques, proteins are identified that associate with 213P1F11 and mediate signaling events. Several pathways known to play a role in cancer biology can be regulated by 213P1F11, including phospholipid pathways such as PI3K, survival pathways such as AKT, NFkB, etc, adhesion and migration pathways, including FAK, Rlio, Rac-1, etc, as well as niitogenic/survival cascades such as ERK, p38, etc (Cell Growth Differ. 2000,11 :279; J Biol Chem. 1999, 274:801; Oncogene. 2000, 19:3003, J. Cell Biol. 1997, 138:913.). Bioinformatic analysis revealed that 213P1F11 can become phosphorylated by serine/threonine as well as tyrosine kinases. Thus, the phosphorylation of 213P1F11 is provided by the present invention to lead to activation of die above listed pathways.

Using, e.g., Western blotting techniques the ability of 213P1F11 to regulate these pathways is confirmed. Cells expressing or lacking 213P1F1 1 are eidier left untreated or stimulated with cytokines, honnones and anti-integrin antibodies. Cell lysates are analyzed using anti-phospho-specific antibodies (Cell Signaling, Santa Cruz Biotechnology) in order to detect phosphorylation and regulation of ERK, p38, AKT, P13K, PLC and odier signaling molecules. When 213P1F11 plays a role in the regulation of signaling pathways, whether individually or communally, it is used as a target for diagnostic, prognostic, preventative and dierapeutic purposes.

To confirm that 213P1F1 1 directly or indirectly activates known signal transduction pathways in cells, luciferase (luc) based transcriptional reporter assays are carried out in cells expressing individual genes. These transcriptional reporters contain consensus-binding sites for known transcription factors that lie downstream of well-characterized signal transduction pathways. The reporters and examples of these associated transcription factors, signal transduction pathways, and activation stimuli are listed below.

NFkB-luc, NFkB/Rel; Ik-kinase/SAP ; growth/apoptosis/stress " SRE-luc, SRF/TCF/ELK1 ; MAPK/SAPK; growth/differentiation AP- l-luc, FOS/JUN; MAPK/SAP /PKC; growth/apoptosis/stress ARE-luc, androgen receptor; sleroids/MAPK; growlli differentiation/apoptosis 115 p53-luc, p53; SAPK; growtli/differentiation/apoptosis CRE-luc, CREB/ATF2; PKA/p38; growtli/apoptosis/stress Gene-mediated effects can be assayed in cells showing mRNA expression. Luciferase reporter plasmids can be introduced by lipid-mediated transfection (TFX-50, Promega). Luciferase activity, an indicator of relative transcriptional activity, is measured by incubation of cell extracts with luciferin substrate and luminescence of the reaction is monitored in a luminometer.

Signaling pathways activated by 213P1F11 are mapped and used for the identification and validation of therapeutic targets. When 213P1F11 is involved in cell signaling, it is used as target for diagnostic, prognostic, preventative and therapeutic purposes.

Example 46: Involvement in Tumor Progression Some apoptosis intermediates, such as DcRl, FL1CE and TRAIL-R3, function as cellular inhibitors of apoptosis by acting as decoys and interfering with normal function of the apoptotic machinery (Sheikh MS et al, Oncogene. 1999, 18:4153; Ashkenazi A, Dixit VM. Curr Opin Cell Biol. 1999, 1 1 :255). When 213P1F1 1 functions as a decoy, it can contribute to the growth of cancer cells. The role of 213P1 F1 1 in tumor growth is confirmed in a variety of primary and transfected cell lines including prostate, bladder and breast cell lines as well as NTH 3T3 cells engineered to stably express 213P1F1 1. Parental cells lacking 213P1F1 1 and cells expressing 213P1F 11 are evaluated for cell growth using a well-documented proliferation assay (Fraser SP, Grimes JA, Djamgoz MB. Prostate. 2000,44:61, Johnson DE, Ochieng J, Evans SL. Anticancer Drugs. 1996, 7:288).

To confirm the role of 213P1F11 in the transformation process, its effect in colony forming assays is investigated. Parental NIH3T3 cells lacking 213P1F11 are compared to NHI-3T3 cells expressing 213P1F1 1 , using a soft agar assay under stringent and more permissive conditions (Song Z. et al. Cancer Res. 2000, 60:6730).

To confirm the role of 213P1F11 in invasion and metastasis of cancer cells, a well-established assay is used, e.g., a Transwell Insert System assay (Becton Dickinson) (Cancer Res. 1999, 59:6010). Control cells, including prostate, colon, bladder and kidney cell lines lacking 213P1F11 are compared to cells expressing 213P1 F1 1. Cells are loaded with the fluorescent dye, calcein, and plated in the top well of the Transwell insert coated with a basement membrane analog. Invasion is detennined by fluorescence of cells in the lower chamber relative to the fluorescence of the entire cell population. 213P1F1 1 can also play a role in cell cycle and apoptosis. Parental cells and cells expressing 213P1F11 are compared for differences in cell cycle regulation using a well-established BrdU assay (Abdel-Malek ZA. J Cell Physiol. 1988, 136:247). In short, cells are grown under both optimal (full serum) and limiting (low serum) conditions are labeled with BrdU mid stained with anti-BrdU Ab and propidium iodide. Cells are analyzed for entry into the Gl, S, and G2M phases of the cell cycle. Alternatively, the effect of stress on apoptosis is evaluated in control parental cells and cells expressing 213P1F11, including normal and tumor bladder cells. Engineered and parental cells are treated with various chemotherapeutic agents, such as paclitaxel, gemcitabine, etc, mid protein synthesis inhibitors, such as cycloheximide. Cells are stained with 116 annexin V-FITC and cell deatli is measured by FACS analysis. The modulation of cell death by 213P1F11 can play a critical role in regulating tumor progression and tumor load.

Whe 213P1F11 plays a role in cell growth, transformation, invasion or apoptosis, it is used as a target for diagnostic, prognostic, preventative mid therapeutic purposes.

Example 47: Involvement in An iouenesis Angiogenesis or new capillary blood vessel formation is necessary for tumor growth (Hanahan D, Folkman J. Cell. 1996, 86:353; Folkman J. Endocrinology. 1998' 139:441). Several assays have been developed to measure angiogenesis in vitro and in vivo, such as the tissue culture assays, endothelial cell tube formation, and endothelial cell proliferation. Using these assays as well as in vitro neo-vascnlarization, die effect of 213P1F11 on angiogenesis is confirmed. For example, endothelial cells engineered to express 213P1F1 1 are evaluated using tube formation and proliferation assays. The effect of 213P1F11 is also confirmed in animal models in vivo. For example, cells either expressing or lacking 213P1 F11 are implanted subcutaneously in immunocompromised mice. Endothelial cell migration and angiogenesis are evaluated 5-15 days later using inmuinohislochemistry techniques. When 213P1F11 affects angiogenesis, it is used as a target for diagnostic, prognostic, preventative and therapeutic purposes Example 48: Regulation of Transcription The localization of 213P1F1 1 to the cytoplasm with potential nuclear localization (Table XXII), support the present invention use of 213P1F11 based on its role in tire transcriptional regulation of eukaryotic genes. Regulation of gene expression is confirmed, e.g., by studying gene expression in cells expressing or lacking 213P1F11. For this purpose, two types of experiments are performed.

In the first set of experiments, RNA from parental and 213P IF 11 -expressing cells are extracted and hybridized to commercially available gene arrays (Clontech) (Smid-Koopman E et al. Br J Cancer. 2000. 83 :246). Resting cells as well as cells treated with FBS or androgen are compared. Differentially expressed genes are identified in accordance with procedures known in the art. The differentially expressed genes are Uien mapped to biological pathways (Chen K et al., Thyroid. 2001. 11:41.).

In the second set of experiments, specific transcriptional pathway activation is evaluated using commercially available (Stratagene) luciferase reporter constructs including: NFkB-luc, SRE-luc, ELKl-luc, ARE-luc, p53-luc, and CRE-luc. These transcriptional reporters contain consensus binding sites for known transcription factors that lie downstream of well-characterized signal transduction pathways, and represent a good tool to ascertain pathway activation mid screen for positive and negative modulators of pathway activation.

When 213P1F11 plays a role in gene regulation, it is used as a target for diagnostic, prognostic, preventative mid therapeutic purposes.

Example 49: Subcellular Localization of 213P1F11 The cellular location of 213P1F11 emi be assessed using subcellular fractionation techniques widely used in cellular biology (Storrie. B, et al. Methods Enzymol. 1990; 182:203-25). A variety of cell lines, including prostate, bladder mid breast cell lines as well as cell lines engineered to express 213P1F1 1 are 117 separated into nuclear, cytosolic and membrane fractions. Gene expression and location in nuclei, heavy membranes (lysosomes, peroxisomes, and mitochondria), light membranes (plasma membrane and endoplasmic reticulum), and soluble protein fractions are tested using Western blotting techniques.

Alternatively, 293T cells can be trausfected with an expression vector' encoding individual genes, HlS-tagged (PCDNA 3.1 MYC/HIS, Invitrogen) mid the subcellular localization of these genes is determined as described above. In short, the trausfected cells can be harvested and subjected to a differential subcellular fractionation protocol (Pemberton, P.A. et al, 1997, J of Histochemistry and Cytochemistry, 45: 1697-1706.). Location of the HIS-tagged genes is followed by Western blotting.

Using 213P1F11 antibodies, it is possible to demonstrate cellular localization by immunofluorescence and immunohislochemistry. For example, cells expressing or lacking 213PIF11 are adhered to a microscope slide and stained with aiiti-213Pl Fl 1 specific Ab. Cells are incubated with an FITC-coupled secondary anti-species Ab, m d analyzed by fluorescent microscopy.

When 213P1F1 1 is localized to specific cell compartments, it is used as a target for diagnostic, preventative and therapeutic purposes.

Example 50: 213PlFi l Proteolytic Activity The similarity of 213P1F11 to casapase cysteine proteases supports tire use of 213P1 F11 as a protease. Protease activity can be confirmed using on in vitro protease assay coupled to detection of protein fragments by western blotting (Hu S et al, above; Slee E et al, J Biol Cliem. 2001, 276:7320). In one embodiment, recombinant 213P1F11 protein is incubated with apoptotic substrates, including other caspases known to associate with caspase 14, namely caspase 2 and caspase 4, as well as recombinant RIP and poly(ADP-ribose) polymerase (i.e. PARP) (Slee E et al, J Biol Chem. 2001, 276:7320; Hayakawa et al, Apoptosis. 2002, 7: 107). Proteins are separated by SDS-Page and analyzed by western blotting with substrate specific antibodies. In another embodiment, 213P1F11 activity is compared in control cells lacking 213P1F1 1 and cells expressing 213P1 F1 1. Cell lysates from control and 213P1F1 1 expressing cells are incubated in the presence of the recombinant substrates listed above. Whole proteins are analyzed by western blotting with antibodies directed to the apoptotic substrates.

When 213P1F11 functions as a protease, it is used as a target for diagnostic, preventative and therapeutic purposes 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.) The 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 Uiose 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 118 modifications or other embodiments can be practiced without departing from the true scope and spirit of the invention. 119 TABLE I: Tissues that Express 213P1F11 When Malignant - Bladder - Prostate - Breast TABLE II: Amino Acid Abbreviations 120 TABLE III: Amino Acid Substitution Matrix Adapted from the GCG Software 9.0 BLOSU 62 amino acid substitution matrix (block substitution matrix). The higher the value, the more likely a substitution is found in related, natural proteins. (See URL at the World Wide Web (.ilq iiube.cli/m nial/blosum62.html ) D E F G H I K L M N P Q R S T V . w Y 2 -1 -2 0 -2 -1 -1 -1 -1 -2 -1 -1 -1 1 0 0 -3 -2 A 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 -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 -3 2 1 -3 -3 -3 -3 -2 -1 3 -3 -1 I -2 -1 0 -1 1 2 0 -1 -2 -3 -2 K 4 2 -3 -3 -2 -2 -2 -1 1 -2 -1 L -2 -2 0 -1 -1 -1 1 -1 -1 M 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 121 TABLE IV HLA Class I/II Motifs/Supermotifs TABLE IV (A): HLA Class I Supei motit's/Motits 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 (B): HLA Class II Supermotif 122 TABLE IV (C): HLA Class Π Motifs MOTIFS . 1° anchor 1 2 4 5 1° anchor 6 DR4 preferred FMYLIVW M T . I VSTCPAUM M deleterious W DR1 preferred MFI/W7 PAMQ VMATSPLIC deleterious C CH FD CWD G DR7 preferred MFZJFPF7 M "W A TVMSACTPL deleterious C G G DR3 MOTIFS 1° anchor 1 2 J 1 ° anchor 4 5 1° anchor 6 motif a LIVMFY D preferred motif b LrVMFAY DNQEST KRH preferred DR MFLIVWY VMSTACPL/ Supermotif Itahcized residues indicate less preferred or "tolerated" residues 123 TABLE IV (D): HLA Class I Supermotifs POSITION: 1 2 4 5 6 7 SUPER- MOTIFS Al 1° Anchor TILVMS P(5/5): (3/5) (4/5) (4/5) G(4/5); A(3/5); QN(3/5) B27 1° Anchor RHK B44 1° Anchor ED B58 1° Anchor ATS B62 Γ Anchor QUI/MP 124 TABLE IV (E): HLA Class I Motifs POSITION: Al preferred GFY l°Anchor DEA YFW P DEQN Y 9-mer W STM deleterious DE RHKLIVMP A Al preferred GRHK ASTCLIVM 1 "Anchor GSTC ASTC LIVM 9-mer OEAS deleterious A RHKDEPY DE PQN RH PG FW Al preferred YFW 1 "Anchor DEAQN A YFWQN PASTC G -mer STM deleterious GP RHKGLIVM DE RHK QNA RHKYFW R Al preferred YFW STCLIVM 1° Anchor YFW -mer DBAS deleterious RHK RHKDEPY P P FW A2.1 preferred YFW 1 "Anchor YFW STC YFW A 9-mer IMIVOAT deleterious DEP DERKH RKH DERKH 125 TABLE IV (E): HLA Class I Motifs, continued POSITION: 1 2 3 4 5 6 7 A2.1 preferred AYFW 1° Anchor LVI G G FY 10-mer IMIVQA V T 1 deleterious DEP DE RKHA P RKH DE A3 preferred PJ-TK 1° Anchor YFW PRHKYFW A YFW LMVISA TFCGD deleterious DEP DE Al l preferred A 1 "Anchor YFW YFW A YFW YFW VTLMIS AGNCDF deleterious DEP A A24 preferred YFW HK 1 "Anchor STC YFW Y 9-mer YFWW deleterious DEG DE G QNP DERH G A K A24 preferred 1 "Anchor P YFWP P -mer YFWM deleterious GDE QN RHK DE A Q A3101 preferred RHK 1 "Anchor YFW P YFW YFW A MVTALIS deleterious DEP DE ADE DE DE D A3301 preferred 1 "Anchor YFW AYFW.

MVALF/ ST deleterious GP DE 126 TABLE rv (E): BDLA Class I Motifs, continued POSITION 1 2 3 4 5 6 7 8 A6801 preferred YFWSTC 1° Anchor YFWLIV YFW P AVTMSLI M deleterious GP DEG RHK A B0702 preferred RH FW 1° Anchor RHK RHK RHK RHK PA Y P deleterious DEQNP DEP DE DE GDE QN DE B3501 preferred FWYLIV 1 "Anchor FWY FWY M P deleterious AGP G G B51 preferred LIVMFW 1 "Anchor FWY STC FWY G FWY Y P deleterious AGPDER DE G DEQN GDE HKSTC B5301 preferred LIVMFW 1° Anchor FWY STC FWY LIVMFWY FWY Y P deleterious AGPQN G RHKQN DE B5401 preferred FWY 1° Anchor FWYL LIVM ALIVM FWY P IVM deleterious GPQNDE GDES RHKDE DE QNDGE" DE TC Itahcized residues indicate less preferred or "tolerated" residues. The information in this Table is specific for 9-mers unless 127 Table V v.l-Al-9mers: 213P1F11 Table V v.l-Al-9mers: 213P1F11 Portion of | SEQID NO: 3; each start position is specified, the length of each peptide is 9| amino acids, the end position for each peptide is the start position plus eight 128 Table Vv.2-Al-9mers: 213P1F11 Table V v.2-Al-9mers: 213P1F11 Pos 123456789 Score 27 PLWNSQDTS 0.000 41 RKAHALSRP 0.000 28 LWNSQDTSP 0.000 55 RRGKDISWN 0.000 11 PFQDPLYLP 0.000 49 PWWMCSRRG 0.000 129 Table V v.3-Al-9mers: 213P1F11 130 Table V v.4-Al-9mers: 213P1F11 Table V v.4-Al-9mers: 213P1F11 Pos 123456789 Score 76 I S FRNSETS 0.002 Portion of 74 LS I.S FRNSE 0.002 SEQ ID 48 HQKLVNDPR 0.002 NO: 9; each start 13 VQPE RTGL 0.002 I position is 28 CGQTFRLKE 0.001 specified, 84 SASEEEKYD 0.001 the length 69 IVGRDLS I S 0.001 of each GLRDENGEC 0.001 "I peptide is 9 amino 33 RLKEEQGRA 0.001 acids, the 40 RAFRGS SVH 0.001 end 65 GVGDIVGRD 0.001 j position for| 32 FRLKEEQGR .0.001 each 59 QEVFGGGVG 0.001 peptide is 61 VFGGGVGDI 0.001 the stait position 78 FRNS ETSAS 0.001 plus eight 67 GDIVGRDLS 0.001 23 DENGECGQT 0.001 79 RNSETSASE 0.001 GKCQEYDKS 0.001 39 GRAFRGSSV 0.001 17 KRTGLRDEN 0.001 38 QGRAFRGS S 0.000 37 EQGRAFRGS 0.000 29 GQTFRLKEE 0.000 41 AFRGS SVHQ 0.000 49 QKLVNDPRE 0.000 DKSLSVQPE 0.000 81 SETSAS EEE 0.000 31 TFRLKEEQG 0.000 77 S RNS ETSA 0.000 53 NDPRETQEV 0.000 19 TGLRDENGE 0.000 YDKSLSVQP 0.000 47 VHQKLVNDP 0.000 16 EKRTGLRDE 0.000 PEKRTGLRD 0.000 131 Table Vv.5-Al-9mers: 213P1F11 jTable Vv.6-Al-9mers: 213P1F11 132 Table VI v.l-Al-10mers: 213P1F11 Table VI v.l-Al-10mers: 213P1F11 133 Table VI v.2-Al-10mers: 213P1F11 Table VI v.2-Al-10mers: 213P1F1I Pos 1234567890 Score Pos 1234567890 Score 34 DTSPTDMIRK 25.000 Portion of 3 VYSTVEGPTP 0.000 Portion of 9 GPTPFQDPLY 2.500 SEQED 55 SRRGKDISWN 0.000 SEQ ID PPNPPL N 0.500 NO: 5; NO: 5; 22 EA 1 LHVYSTVEGP 0.000 each start each start 6 , TVEGPTPFQD 0.450 position is 51 WWMCSRRGKD 0.000 position is PSEAPPNPPL 0.270 specified, 28 PLWNSQDTSP 0.000 specified, 37 PTDMIRKAHA 0.250 the length 41 IRKAHALSRP 0.000 the length 47 LSRPWWMCSR 0.150 of each of each 4 YSTVEGPTPF 0.150 peptide is peptide is amino 10 amino 13 FQDPLYLPSE 0.150 acids, the acids, the 32 SQDTSPTDMI 0.075 end end STVEGPTPFQ . 0.050 position for position for 33 QDTSPTDMIR 0.025 each each 43 KAHALSRPWW 0.020 peptide is peptide is NSQDTSPTDM 0.015 the start 31 tire start position position TSPTDMIRKA 0.015 plus nine plus nine PTPFQDPLYL 0.013 45 HALSRPWWMC 0.010 21 SEAPPNPPLW 0.010 17 LYLPSEAPPN 0.010 2 HVYSTVEGPT 0.010 40 MIRKAHALSR 0.005 52 WMCSRRGKDI 0.005 46 ALSRPWWMCS 0.005 48 SRPWWMCSRR 0.005 8 EGPTPFQDPL 0.003 26 NPPLWNSQDT 0.003 24 PPNPPLWNSQ 0.003 36 SPTDMIRKAH 0.003 23 APPNPPLWNS 0.003 18 YLPSEAPPNP 0.002 39 DMIRKAHALS 0.001 53 MCSRRGKDIS 0.001 54 CSRRGKDISW 0.001 56 RRGKDISWNF 0.001 42 RKAHALSRPW 0.001 44 AHALSRPWWM 0.001 14 QDPLYLPSEA 0.001 11 TPFQDPLYLP 0.001 38 TDMIRKAHAL 0.001 29 LWNSQDTSPT 0.001 7 VEGPTPFQDP 0.001 WNSQDTSPTD 0.001 12 PFQDPLYLPS 0.000 27 PPLWNSQDTS 0.000 19 LPSEAPPNPP 0.000 DPLYLPSEAP 0.000 PNPPLWNSQD 0.000 49 RPWWMCSRRG 0.000 16 PLYLPSEAPP 0.000 50 P WMCSRRGK 0.000 135 Table VI v.4-Al-10mers: 213P1F11 Table VI v.4-Al-10mers: 213P1F11 Pos 1234567890 Score Pos 1234567890 Sco e 85 ASEEEKYDMS 1.350 Portion of 29 GQTFRLKEEQ 0.002 Portion of 82 ETSASEEEKY 1.250 SEQID 41 AFRGSSVHQK 0.001 SEQID 52 VNDPRETQEV 1.250 NO: 9; 20 GLRDENGECG 0.001 NO: 9; each start each start NGECGQTFRL 1.125 position is 73 DLSISFRNSE 0.001 position is 34 LKEEQGRAFR 0.900 specified, 51 LVNDPRETQE 0.001 specified, 4 CQEYDKSLSV 0.675 the length 46 SVHQKLVNDP 0.001 the length KEEQGRAFRG 0.225 of each 47 VHQKLVNDPR 0.001 of each 86 SEEEKYDMSG 0.225 peptide is 31 TFRLKEEQGR 0.001 peptide is amino KSLSVQPEKR 0.150 53 NDPRETQEVF 10 amino 9 acids, the 0.001 acids, the 80 NSETSASEEE 0.135 end 49 QKLVNDPRET .0.001 end 58 TQEVFGGGVG 0.135 position for 2 GKCQEYDKSL 0.001 position for 66 VGDIVGRDLS 0.125 each 56 RETQEVFGGG 0.001 each 71 GRDLSISFRN 0.125 peptide is 6 EYDKSLSVQP 0.001 peptide is 100 the stiirt 12 SVQPEKRTGL 0. 54 DPRETQEVFG 0.001 the start position position 44 GSSVHQKLVN 0.075 plus nine 39 GRAFRGSSVH 0.001 plus nine 63 GGGVGDIVGR 0.050 42 FRGSSVHQKL 0.001 69 IVGRDLSISF 0.050 61 VFGGGVGDIV 0.001 22 RDENGECGQT 0.045 5 QEYDKSLSVQ 0.001 57 ETQEVFGGGV ' 0.025 72 RDLSISFRNS 0.001 24 ENGECGQTFR 0.025 17 KRTGLRDENG 0.001 21 LRDENGECGQ 0.025 36 EEQGRAFRGS 0.001 55 PRETQEVFGG .0.023 79 RNSETSASEE 0.000 84 SASEEEKYDM 0.020 1 MGKCQEYDKS 0.000 8 DKSLSVQPEK 0.020 38 QGRAFRGSSV 0.000 11 LSVQPEKRTG 0.015 19 TGLRDENGEC 0.000 13 VQPEKRTGLR 0.015 64 GGVGDIVGRD 0.000 74 LSISFRNSET 0.015 28 CGQTFRLKEE 0.000 62 EGGGVGDIVG 0.013 37 EQGRAFRGSS 0.000 14 QPEKRTGLRD 0.011 78 FRNSETSASE 0.000 SLSVQPEKRT 0.010 59 QEVFGGGVGD 0.000 60 EVFGGGVGDI 0.010 77 SFRNSETSAS 0.000 75 SISFRNSETS 0.010 16 EKRTGLRDEN 0.000 65 GVGDIVGRDL 0.010 32 FRLKEEQGRA . 0.000 3 KCQEYDKSLS 0.010 48 HQKLV DPRE 0.000 68 DIVGRDLSIS 0.010 7 YDKSLSVQPE 0.000 81 SETSASEEEK 0.010 15 PEKRTGLRDE 0.000 26 GECGQTFRLK 0.010 33 RLKEEQGRAF 0.010 50 KLV DPRETQ 0.010 23 DENGECGQTF 0.005 27 ECGQTFRLKE 0.005 45 SSVHQKLVND 0.003 67 GDIVGRDL'SI 0.003 70 VGRDLSISFR 0.003 QTFRLKEEQG 0.003 18 RTGLRDENGE 0.003 43 ■ RGSSVHQKLV 0.003 40 RAFRGSSVHQ 0.002 83 TSASEEEKYD 0.002 76 ISFRNSETSA 0.002 136 Table VI v.5-Al-10mers: 213P1F11 Table VI v.6-Al-10mers: 213P1F11 137 Table VII v.l-A2-9mers: 213P1F11 Table VII v.l-A2-9mers: 213P1F11 138 Table VTI v.2-A2-9mers: 213P1F11 Table VII v.2-A2-9mers: 213P1F11 Pos 123456789 Score 28 LWNSQDTSP 0.000 Portion of 11 PFQDPLYLP 0.000 SEQID 54 SRRGKDISW 0.000 NO: 5; each start 49 PW MCSRRG 0.000 position is j 19 PSEAPPNPP 0.000 specified, 40 IRKAHALS 0.000 the length of each peptide is 9[ amino acids, the end position for| each peptide is the start position plus eight 139 Table VII v.3-A2-9mers: 213P1F11 140 Table VII v.4-A2-9mers: 213P1F11 Table VII v.4-A2-9mers: 213P1F11 Pos 123456789 Score Pos 123456789 Score QEYDKSLSV 17.743 Portion of 63 GGGVGDIVG 0.000 Portion of 13 VQPEKRTGL 15.096 SEQID 45 SSVHQKLVN 0.000 SEQID KLVNDPRET 5.216 NO: 9; 50 24 ENGECGQTF 0.000 NO: 9, each start each start 26 GECGQTFRL • 2.409 position is 81 SE SASEEE 0.000 position is 3 KCQEYDKSL 2.001 specified, 57 ETQEVFGGG 0.000 specified, 75 SISFRNSET 1.025 the length 59 QEVFGGGVG 0.000 the length 44 GSSVHQKLV 0.454 of each 49- QKLVNDPRE 0.000 of each 62 FGGGVGDIV 0.420 peptide is 9 2 GKCQEYDKS 0.000 peptide is 9 amino amino 58 TQEVFGGGV 0.223 52 V DPRETQE acids, the 0.000 acids, the GLRDENGEC 0.201 end 67 GDIVGRDLS 0.000 end 43 RGSSVHQKL 0.139 position for 78 FRNSETSAS 0.000 position for 68 DIVGRDLSI 0.108 each 47 VHQKLVNDP 0.000 each 53 NDPRETQEV 0.097 peptide is 32 FRLKEEQGR 0.000 peptide is the start 33 . RLKEEQGRA 0.093 25 NGECGQTFR the start 0.000 position position 11 LSVQPEKRT 0.083 plus eight 42 FRGSSVHQK 0.000 plus eight 56 RETQEVFGG 0.019 38 QGRAFRGSS. 0.000 66 VGDIVGRDL 0.019 17 KRTGLRDEN 0.000 69 IVGRDLSIS 0.010 21 LRDENGECG 0.000 39 GRAFRGSSV 0.010 71 GRDLSISFR 0.000 85 ASEEEKYDM 0.009 34 LKEEQGRAF 0.000 SLSVQPEKR 0.007 ' 82 ETSASEEEK 0.000 84 SASEEEKYD 0.005 80 NSETSASEE 0.000 51 LV DPRETQ 0.004 1 MGKCQEYDK 0.000 61 VFGGGVGDI 0.004 8 DKSLSVQPE 0.000 29 GQTFRLKEE 0.003 7 YDKSLSVQP 0.000 46 SVHQKLVND 0.003 27 ECGQTFRL 0.000 72 RDLSISFRN 0.002 54 DERETQEVF 0.000 73" DLSISFRNS 0.002 31 TFRLKEEQG 0.000 65 GVGDIVGRD 0.002 22 RDENGECGQ 0.000 40 RAFRGSSVH 0.002 48 HQKLVNDPR 0.000 76 ISFRNSETS 0.001 14 QPEKRTGLR . 0.000 23 DENGECGQT 0.001 41 AFRGSSVHQ 0.000 12 SVQPEKRTG 0.001 15 PEKRTGLRD 0.000 9 KSLSVQPEK 0.001 55 PRETQEVFG 0.000 36 EEQGRAFRG 0.001 6 EYDKSLSVQ 0.000 . 74 LSISFRNSE 0.001 16 EKRTGLRDE 0.000 18 RTGLRDENG 0.001 4 CQEYDKSLS 0.000 79 RNSETSASE 0.000 QTFRLKEEQ 0.000 28 CGQTFRLKE ' 0.000 60 EVFGGGVGD 0.000 19 TGLRDENGE 0.000 KEEQGRAFR 0.000 86 SEEEKYDMS 0.000 77 SFRNSETSA 0.000 . 70 VGRDLSISF 0.000 83 TSASEEEKY 0.000 64 GGVGDIVGR 0.000 37 EQGRAFRGS 0.000 Table Vil v.5-A2-9mers: 213P1F11 Table VII v.6-A2-9mers: 213P 1F11 142 Table VIII v. l-A2-10mers: 213P1F11 Table VIII v. l-A2-10mers: 2I3P1F11 Pos 1234567890 Score Pos 1234567890 Score 187 FIQTLVDVFT 25.924 Portion of 15 SGARLALI LC 0.075 Portion of | 11 1 ALNNKNCQAL 21.362 SEQ 1D 165 ALHVYSTVEG 0.075 SEQ ID LMAHGREGFL 18.753 NO: 3; 86 125 KVYI IQACRG 0.073 NO: 3; each start each start 142 TVGGDEIVMV 13.997 position is 167 HVYSTVEGYI 0.071 position is | 149 VMVI KDS PQT 9.149 specified, 104 KLENLFEALN 0.063 specified, 117 CQALRAKPKV 7.052 the length 56 PTAEQFQEEL 0.050 the length 205 LLTEVTRRMA 6.925 of each 30 EGSEEDLDAL 0.048 of each 94 FLKGEDGEMV 5.487 peptide is 175 YIAYRHDQKG 0.047 peptide is amino PKVYI 41 HMFRQLRFES 10 amino 119 ALRAK 4.361 acids, the 0.039 acids, the 185 SCFIQTLVDV 3.864 end 209 VTRRMAEAEL 0.038 end 75 REDPVSCAFV 2.975 position for 188 IQTLVDVFTK 0.034 position for| 70 QAI DSREDPV 1.941 each 193 DVFTKRKGHI 0.033 each 183 KGSCFIQTLV 1.589 peptide is 10 EKYDMSGARL 0.029 peptide is the start MVI KDS PQTI 1.552 22 I LCVTKAREG 0.025 the start 150 position position 13 DMSGARLALI 1.300 plus nine 225 RKTNPEIQST 0.024 plus nine 107 NLFEALNNKN 1.130 154 DS PQTI PTYT 0.020 226 KTNPEI QSTL 1.038 110 EALNNKNCQA 0.019 19 LALI LCVTKA 0.998 76 EDPVSCAFW 0.017 218 LVQEGKARKT 0.909 156 PQTI PTYTDA 0.017 63 EELEKFQQAI 0.877 179 RHDQKGSCFI 0.016 159 I PTYTDALHV 0.772 122 AKPKVYI IQA 0.016 232 QSTLRKRLYL 0.767 20 ALI LCVTKAR 0.015 103 . VKLENLFEAL 0.712 18 RLALI LCVTK 0.015 134 GEQRDPGETV 0.663 - 222 GKARKTN PEI 0.014 12 YDMSGARLAL 0.505 6 SLEEEKYDMS 0.014 RSLEEEKYDM 0.492 21 LI LCVTKARE 0.013 143 VGGDEIVMVI 0.448 43 FRQLRFESTM 0.012 162 YTDALHVYST 0.438 62 QEELEKFQQA 0.012 102 MVKLENLFEA 0.345 72 I DSREDPVSC 0.012 48 FESTMKRDPT 0.327 170 STVEGYIAYR 0.011 96 KGEDGEMVKL 0.295 61 FQEELEKFQQ 0.01 1 204 ELLTEVTRRM 0.276 198 RKGHI LELLT 0.010 140 GETVGGDEIV 0.272 194 VFTKRKGHI L 0.010 182 QKGSCFIQTL 0.259 158 TI PTYTDALH 0.010 190 TLVDVFTKRK 0.232 123 KPKVYI IQAC 0.009 85 VLMAHGREGF 0.230 81 CAFWLMAHG 0.009 207 TEVTRRMAEA 0.222 148 IVMVI KDS PQ . 0.008 230 EIQSTLRKRL 0.220 84 WLMAHGREG 0.008 16 GARLALI LCV 0.169 77 DPVSCAFWL 0.008 27 KAREGS EEDL . 0.159 152 I KDS PQTI PT 0.007 157 QTI PTYTDAL 0.145 176 IAYRHDQKGS 0.006 163 TDALHVYSTV 0.145 44 RQLRFESTMK 0.006 37 DALEHMFRQL 0.128 100 GEMVKLENLF 0.005 79 VSCAFWLMA 0.127 196 TKRKGHI LEL 0.005 200 GHI LELLTEV 0,11 1 101 EMVKLENLFE 0.004 201 HI LELLTEVT 0.106 203 LELLTEVTRR 0.004 212 RMAEAELVQE 0.102 181 DQKGSCFIQT 0.004 14 MSGARLALI L 0.097 38 ALEHMFRQLR 0.004 29 REGS EEDLDA 0.097 17 ARLALI LCVT 0.004 78 PVSCAFWLM 0.084 169 YSTVEGYIAY 0.003 143 Table VIII v.2-A2-10mers: 213P1F11 Table Vlll v.2-A2-10mers: 213P1F1 1 Pos 1234567890 Score 24 P PNPPLWNSQ 0.000 Portion of VYSTVEGPTP 0.000 SEQ ID PN PPLWNSQD 0.000 NO: 5; each start 48 SRPWWMCSRR 0.000 position is 41 I RKAHALSRP 0.000 specified, 50 PWWMCSRRGK 0.000 the length of each peptide is 10 amino acids, the end J position for| each peptide is the- staii position plus nine 144 145 TableVIU v.4-A2-10mers: 213P1F11 TableVIU v.4-A2-10mers: 213P1F11 Pos 1234567890 Score Pos 1234567890 Score SLSVQPEKRT 7.452 Portion of 11 LSVQPEKRTG 0.000 Portion of 12 SVQPEKRTGL 1.869 SEQ ID 24 ENGECGQTFR 0.000 SEQ ID NO: 9; 65 GVGDIVGRDL 1.533 66 VGDIVGRDLS 0.000 NO: 9; each start each stail 43 RGSSVHQKLV 0.454 position is 72 RDLSISFRNS 0.000 position is 4 CQEYDKSLSV 0.451 specified, 81 SETSASEEEK 0.000 specified, 52 VNDPRETQEV 0.309 the length 26 GECGQTFRLK 0.000 the length 84 SASEEEKYDM 0.283 of each 23 DENGECGQTF 0.000 of each 76 ISFRNSETSA 0.204 peptide is 85 ASEEEKYDMS 0.000 peptide is amino TQEVFGGGV 22 RDENGECGQT 10 amino 57 E 0.147 acids, the 0.000 acids, the 74 LSISFRNSET 0.083 end 54 DPRETQEVFG 0.000 end 60 EVFGGGVGDI 0.076 position for 34 LKEEQGRAFR 0.000 position for NGECGQTFRL 0.052 each 36 EEQGRAFRGS 0.000 each 38 QGRAFRGSSV 0.035 peptide is 82 ETSASEEEKY 0.000 peptide is . the start D 0.000 the start 2 GKCQEYDKSL 0.030 64 GGVGDIVGR position position 50 KLVNDPRETQ 0.026 plus nine 27 ECGQTFRLKE 0.000 plus nine 61 VFGGGVGDIV 0.016 58 TQEVFGGGVG 0.000 19 TGLRDENGEC 0.016 71 GRDLSISFRN 0.000 67 GDIVGRDLSI 0.014 1 MGKCQEYDKS 0.000 42 FRGSSVHQKL 0.014 53 NDPRETQEVF 0.000 GLRDENGECG 0.011 17 KRTGLRDENG 0.000 69 IVGRDLSISF 0.011 78 FRNSETSASE 0.000 51 LVNDPRETQE 0.009 47 VHQKLVNDPR 0.000 49 QKLVNDPRET 0.008 7 YDKSLSVQPE 0.000 3 KCQEYDKSLS 0.007 14 QPEKRTGLRD 0.000 75 SISFRNSETS 0.005 21 LRDENGECGQ 0.000 73 DLSISFRNSE 0.004 39. GRAFRGSSVH 0.000 QEYDKSLSVQ 0.004 . 77 SFR SETSAS 0.000 46 SVHQKLVNDP 0.003 80 NSETSASEEE 0.000 33 RLKEEQGRAF 0.002 41 AFRGSSVHQK 0.000 KEEQGRAFRG 0.002 48 HQKLV DPRE 0.000 QTFRLKEEQG 0.002 8 DKSLSVQPEK 0.000 32 FRLKEEQGRA 0.002 31 TFRLKEEQGR 0.000 13 VQPEKRTGLR 0.001 16 EKRTGLRDEN 0.000 62 FGGGVGDIVG 0.001 55 PRETQEVFGG . 0.000 40 RAFRGSSVH.Q 0.001 15 • PEKRTGLRDE 0.000 29 GQTFRLKEEQ 0.001 6 EYDKSLSVQP 0.000 - 68 DIVGRDLSIS 0.001 70 VGRDLSISFR 0.001 9 KSLSVQPEKR 0.001 83 TSASEEEKYD 0.001 79 RNSETSASEE 0.000 86 SEEEKYDMSG 0.000 56 RETQEVFGGG 0.000 59 QEVFGGGVGD 0.000 37 EQGRAFRGSS 0.000 45 SSVHQKLV D 0.000 28 CGQTFRLKEE 0.000 63 GGGVGDIVGR 0.000 18 RTGLRDENGE 0.000 44 GSSVHQKLVN 0.000 146 TabLe VIII v.5-A2-10mers: 213P1F11 Table VIII v.6-A2-10mers: 213P1FI1 147 Table IX v.l-A3-9mers: 213P1F11 Tabie IX v.l-A3-9mers: 213P1F1] Portion of SEQID NO: 3; each start position is specified, the length of each peptide is 9| amino ' acids, the end position for| each peptide is the start position plus eight 148 Table IX v.2-A3-9mers: 213P1F11 Table IX v.2-A3-9mers: 213P1F11 Pos 123456789 Score Pos 123456789 Score 45 ALSRPWWMC 0.900 Portion of 7 EGPTPFQDP 0.000 Portion of 34 TSPTDMIRK .0.300 SEQ ID 28 LWNSQDTSP 0.000 SEQ ID NO: 0.000 NO: MIRKAHAL 0.270 5; 5; 38 D 19 PSEAPPNPP each stiirt each start 4 STVEGPTPF 0.225 PFQDPLYLP position is 11 0.000 position is 48 RPWWMCSRR 0.200 specified, 24 PNPPLWNSQ 0.000 specified, 33 DTSPTDMIR 0.180 the length 49 PWWMCSRRG 0.000 the length 8 GPTPFQDPL 0.081 of each of each TPFQDPLYL 0.060 peptide is 9 peptide is 9 amino amino 1 ■ HVYSTVEGP 0.030 acids, the acids, the 17 YLPSEAPPN 0.020 end end 27 PLWNSQDTS 0.020 position for position for 9 PTPFQDPLY 0.020 each each 47 SRPWWMCSR 0.018 peptide is peptide is YLPSEAP 0.015 the start the start PL position position 56 RGKDISWNF 0.009 plus eight plus eight 44 HALS PWWM 0.009 40 IRKAHALSR 0.008 51 WMCSRRGKD 0.006 31 SQDTSPTDM 0.006 TVEGPTPFQ 0.005 SEAPPNPPL 0.004 39 MIRKAHALS 0.004 12 FQDPLYLPS 0.004 50 WWMCSRRGK 0.003 52 MCSRRGKDI 0.003 46 LSRPWWMCS 0.002 32 QDTSPTDMI 0.001 14 DPLYLPSEA 0.001 21 EAPPNPPLW 0.001 36 PTDMIRKAH 0.001 22 APPNPPLWN 0.001 42 KAHALSRPW 0.001 NPPLWNSQD 0.001 54 SRRGKDISW 0.001 23 PPNPPL NS 0.000 18 LPSEAPPNP 0.000 37 TDMI RKAHA 0.000 SPTDMIRKA ■ 0.000 6 VEGPTPFQD 0.000 2 WNSQDTSPT 0.000 43 AHALSRPWW 0.000 53 CSRRGKDIS 0.000 26 PPLWNSQDT 0.000 3 YSTVEGPTP 0.000 NSQDTSPTD . 0.000 13 QDPLYLPSE 0.000 16 LYLPSEAPP- 0.000 2 VYSTVEGPT 0.000 55 RRGKDISWN 0.000 41 RKAHALSRP 0.000 Table IX v.3-A3-9mers: 213P1F11 150 Table IX v.4-A3-9mers: 213P1F11 Table IX v.4-A3-9mers: 2I3P1FU Pos 123456789 Score Pos 123456789 Score SLSVQPEKR 4.000 Portion of 34 LKEEQGRAF 0.000 Portion of 9 KSLSVQPEK 0.675 SEQID 45 SSVHQKLVN 0.000 SEQID NO: 9; 82 ETSASEEEK 0.300 53 NDPRETQEV 0.000 NO: 9; each start eacli start 48 HQKLVNDPR 0.180 position is 77 SFRNSETSA 0.000 position is GLRDENGEC 0.180 specified, 67 GDIVGRDLS 0.000 specified, 33 RL EEQGRA 0.090 the length 86 SEEEKYDMS 0.000 the length. 68 DIVGRDLSI 0.081 of each 84 SASEEEKYD 0.000 of each 42 FRGSSVHQK 0.060 peptide is 9 72 RDLSISFRN 0.000 peptide is 9 amino amino 1 MGKCQEYDK 0.060 2 GKCQEYDKS acids, the 0.000 acids, the 50 KLVNDPRET 0.045 end 63 GGGVGDIVG 0.000 end 64 GGVGDIVGR 0.041 position for 28 CGQTFRLKE 0.000 position for 3 KCQEYDKSL 0.041 each 37 EQGRAFRGS 0.000 eacli KEEQGRAFR 0.036 peptide is 80 NSETSASEE 0.000 peptide is the start 13 VQPEKRTGL 0.027 66 VGDIVGRDL 0.000 the start position position 26 GECGQTFRL 0.024 plus eight 17 KRTGLRDEN 0.000 plus eight 83 TSASEEEKY 0:020 36 EEQGRAFRG 0.000 71 GRDLSISFR 0.018 52 VNDPRETQE 0.000 27 ECGQTFRLK 0.018 79 RNSETSASE 0.000 14 QPEKRTGLR 0.012 47 VHQKLVNDP 0.000 40 RAFRGSSVH 0.010 81 SETSASEEE 0.000 75 SISFRNSET . 0.010 23 DENGECGQT 0.000 54 DPRETQEVF 0.009 78 FR SETSAS 0.000 65 GVGDIVGRD 0.008 38 QGRAFRGSS 0.000 32 FRLKEEQGR 0.006 21 LRDENGECG 0.000 69 IVGRDLSIS 0.006 19 TGLRDENGE 0.000 QEYDKSLSV 0.006 49 QKLVNDPRE 0.000 58 TQEVFGGGV 0.005 41 AFRGSSVHQ 0.000 QTFRLKEEQ 0.005 9 QEVFGGGVG 0.000 85 ASEEEKYDM 0.00 31 TFRLKEEQG 0.000 60 EVFGGGVGD 0.005 7 YDKSLSVQP 0.000 70 VGRDLSISF 0.004 22 RDENGECGQ 0.000 NGECGQTFR 0.004 8 DKSLSVQPE 0.000 73 DLSISFRNS 0.004 15 PEKRTGLRD 0.000 51 LVNDPRETQ 0.003 6 EYDKSLSVQ 0.000 46 SVHQKLVND 0.003 55 PRETQEVFG 0.000 24 ENGECGQTF 0.002 16 EKRTGLRDE 0.000 44 GSSVHQKLV 0.002 29 GQTFRLKEE 0.001 4 CQEYDKSLS 0.001 18 RTGLRDENG 0.001 76 ISFRNSETS 0.001 43 RGSSVHQKL 0.001 61 VFGGGVGDI 0:001 57 ETQEVFGGG. 0.001 39 GRAFRGSSV 0.001 11 LSVQPEKRT 0.001 56 RETQEVFGG 0.001 62 FGGGVGDIV 0.000 74 LSISFRNSE 0.000 12 SVQPEKRTG 0.000 !TabJe IX v.5-A3-9mers: 213P1F11 Table IXv.6-A3-9mers: 213P1F11 152 Table X v. l-A3-10mers Table X v. l -A3- lOmers Pos 1234567890 Score Pos ' 1234567890 Score 190 TLVDVFTKRK 45.000 Portion of 51 TMKRDPTAEQ 0.030 Portion of 18 RLALILCVTK 20.000 SEQ K) 209 VTRRMAEAEL 0.030 SEQ 1D 38 ALEHMFRQLR 12.000 NO: 3; 193 DVFTKRKGHI 0.027 NO: 3; each start each stail 217 ELVQEGKARK 9.000 position is 153 KDSPQTIPTY. 0.027 position is 45 QLRFESTMKR 8.000 specified, 106 ENLFEALNNK 0.027 specified, 188 IQTLVDVFTK 5.400 the length 27 AREGSEEDL 0.027 the length ALILCVTKAR 4.500 of each 100 •GEMVKLENLF 0.027 o each 202 ILELLTEVTR 4.000 peptide is 201 HILELLTEVT 0.022 peptide is amino 85 VLMAHGREGF . 3.000 158 TI PTYTDALH 10 amino acids, the 0.020 acids, the DLDALEHMFR 2.400 end 165 ALHVYSTVEG 0.020 end 170 STVEGYIAYR 2.025 position for 166 LHVYSTVEGY 0.018 position for 189 QTLVDVFTKR 1.350 each 16 GARLALILCV 0.018 each 87 MAHGREGFLK 0.900 peptide is 78 PVSCAFWLM 0.018 peptide is LRFESTMK the start 44 RQ 0.900 101 EMVKLENLFE 0.018 the start position position 119 ALRAKPKVYI 0.900 plus nine 187 FIQTLVDVFT 0.015 plus nine 41 HMFRQLRFES 0.600 185 SCFIQTLVDV 0.015 11 ALNNKNCQAL 0.600 123 KPKVYIIQAC 0.013 13 DMSGARLALI 0.405 141 ETVGGDEIVM 0.013 128 IIQACRGEQR 0.400 204 ELLTEVTRRM 0.013 228 NPEIQSTLRK 0.400 56 PTAEQFQEEL 0.013 94 FLKGEDGEMV 0.300 231 IQSTLRKRLY 0.012 144 GGDEIV VIK 0.203 . 227 TNPEIQSTLR 0.012 157 QTI PTY DAL 0.203 39 LEHMFRQLRF 0.012 226 ' KTNPEIQSTL 0.203 186 CFIQTLVDVF 0.009 86 LMAHGREGFL 0.180 79 VSCAFWLMA 0.009 104 KLENLFEALN 0.180 66 EKFQQAIDSR 0.009 107 NLFEALNNKN 0.150 216 AELVQEGKAR 0.009 160 PTYTDALHVY 0.150 19 LALILCVTKA 0.009 149 VMVIKDSPQT 0.150 80 SCAFWLMAH 0.009 59 EQFQEELEKF- 0. 135 230 EIQSTLRKRL 0.009 214 AEAELVQEGK 0.135 77 DPVSCAFWL 0.008 167 HVYSTVEGYI 0.135 181 DQKGSCFIQT 0.008 171 TVEGYIAYRH 0.135 112 LN KNCQALR 0.008 58 AEQFQEELEK 0.120 233 STLRKRLYLQ 0.007 , 1 16 NCQALRAKPK 0.100 5 RSLEEEKYDM • 0.007 174 GYIAYRHDQK 0.090 197 KRKGHILELL 0.006 150 MVIKDSPQTI 0.090 82 AFWLMAHGR 0.006 102 VKLENLFEA 0.090 14 MSGARLALIL 0.006 95 LKGEDGEMVK 0.060 232 QSTLRKRLYL 0.006 6 SLEEEKYDMS 0.060 1 17 CQALRAKPKV 0.006 203 LELLTEVTRR 0.054 64 ELEKFQQAID 0.006 162 YTDALHVYST 0.045 143 VGGDEIVMVI 0.005 142 TVGGDEIVMV 0.045 120 LRAKPKVYII 0.005 212 RMAEAELVQE 0.045 103 VKLENLFEAL 0.004 169 YSTVEGYIAY 0.040 159 I PTYTDALHV 0.004 2 SNPRSLEEEK 0.040 71 AIDSREDPVS 0.004 3 NPRSLEEEKY 0.040 63 EELEKFQQAI 0.004 1 18 QALRAKPKVY 0.030 74 SREDPVSCAF 0.003 125 KVYIIQACRG 0.030 114 NKNCQALRAK 0.003 205 LLTEVTRRMA 0.030 148 IV VIKDSPQ 0.003 Table Xv.2-A3-10mers: 213P1F11 Table X v.2-A3-10mers: 213P1F11 Pos 1234567890 Score Pos 1234567890 Score 34 DTSPTDMIRK 1.350 Portion of 41 IRKAHALSRP 0.000 Portion of 40 MIRKAHALSR 0.800 SEQID 3 VYSTVEGPTP 0.000 SEQ ID 2 WMCSRRGKDI 0.300 NO: 5; 42 RKAHALSRPW 0.000 NO: 5; each start each start 46 ALSRP WMCS 0.240 position is 25 PNPPL NSQD 0.000 position is 9 GPTPFQDPLY 0.180 specified, 51 WWMCSRRGKD 0.000 specified, 47 LSRPWWMCSR 0.135 the length 12 PFQDPLYLPS 0.000 the length 32 SQDTSPTDMI 0.027 of each of each 2 HVYSTVEGPT 0.022 peptide is peptide is amino YLPSEAPPNP 10 amino 18 0.020 acids, the acids, the 39 DMIRKAHALS 0.018 end end 45 HALSRPWWMC 0.013 position for position for 16 PLYLPSEAPP 0.010 each each 4 YSTVEGPTPF 0.010 . peptide is peptide is PLWNSQDTSP 0.010 the start the start 28 position position 56 RRGKDI SWNF 0.009 plus nine plus nine 6 TVEGPTPFQD 0.009 33 QDTSPTDMIR 0.008 11 . TPFQDPLYLP 0.007 PTPFQDPLYL 0.006 43 KAHALSRPWW 0.006 13 FQDPLYLPSE 0.004 48 SRPWWMCSRR 0.004 STVEGPTPFQ 0.003 23 APPNPPL NS 0.003 54 CSRRGKDIS 0.002 36 SPTDMIRKAH 0.002 50 PWWMCSRRGK 0.001 31 NSQDTSPTDM 0.001 37 PTDMIRKAHA 0.001 26 NPPLWNSQDT 0.001 38 TDMIRKAHAL 0.001 21 SEAPPNPPLW 0.001 44 AHALSRPW M 0.001 8 EGPTPFQDPL 0.001 PSEAPPNPPL 0.000 19 LPSEAPPNPP 0.000 7 VEGPTPFQDP 0.000 53 MCSRRGKDIS 0.000 22 EAPPNPPLWN 0.000 14 QDPLYLPSEA 0.000 TSPTDMIRKA 0.000 DPLYLPSEAP 0.000 29 LWNSQDTSPT 0.000 49 RPWWMCSRRG 0.000 1 LHVYSTVEGP 0.000 27 PPLWNSQDTS 0.000 55 SRRGKDISWN 0.000 17 LYLPSEAPPN 0.000 WNSQDTSPTD 0.000 24 PPNPPLWNSQ 0.000 Table X v.3-A3-10mers: 213P1F11 Pos 1234567890 Score 11 TLPSPFPYLS 0.360 Portion of 9 GATLPSPFPY 0.360 SEQ 113 ATLPSPFPYL 0.304 NO: 7; each start 3 IIQACRGATL 0.060 position is 12 LPSPFPYLSL 0.027 specified, 2 YIIQACRGAT 0.005 the length 7 CRGATLPSPF 0.002 of each QACRGATLPS 0.001 peptide is amino 4 IQACRGATLP 0.001 . acids, the 6 ACRGATLPSP 0.000 end 8 RGATLPSPFP 0.000 position for 1 VYIIQACRGA 0.000 each peptide is the start position plus nine Table X v.4-A3-10mers: 213P1F11 Table Xv.4-A3-10mers: 213P1F11 Pos 1234567890 Score Pos 1234567890 Score 69 IVGRDLSISF 0.400 Portion of 14 QPEKRTGLRD 0.000 Portion of 33 RLKEEQGRAF 0.300' SEQED 27 ECGQTFRLKE 0.000 SEQED KLVNDPRETQ 0.135 NO: 9; 37 EQGRAFRGSS 0.000 NO: 9; 50 each start each stmt 60 EVFGGGVGDI 0.121 position is 85 ASEEEKYDMS 0.000 position is 41 AFRGSSVHQK 0.090 specified, 71 GRDLSISFRN 0.000 specified, 12 SVQPEKRTGL 0.090 the length 45 ' SSVHQKLVND 0.000 the length 9 KSLSVQPEKR 0.090 of each 38 QGRAFRGSSV 0.000 of each 26 GECGQTFRLK 0.081 peptide is 64 GGVGDIVGRD 0.000 peptide is amino 1 VQPEKRT 0.075 58 TQEVFGGGVG 0 amino SLS acids, the 0.000 acids, the GLRDENGECG 0.060 end 80 NSETSASEEE 0.000 end 81 SETSASEEEK 0.060 position for '43 RGSSVHQKLV 0.000 position for 82 ETSASEEEKY 0.060 each 17 KRTGLRDENG 0.000 each 13 VQPEKRTGLR 0.054 peptide is 32 FRLKEEQGRA 0.000 peptide is GVGDIVGRDL 0.027 the stail. 65 19 TGLRDENGEC 0.000 the start position position 63 GGGVGDIVGR 0.018 plus nine 54 DPRETQEVFG 0.000 plus nine 73 DLSISFRNSE 0.018 59 QEVFGGGVGD 0.000 4 CQEYDKSLSV 0.012 56 RETQEVFGGG 0.000 84 SASEEEKYDM 0.009 79 RNSETSASEE 0.000 8 DKSLSVQPEK 0.009 62 FGGGVGDIVG 0.000 70 VGRDLSISFR 0.006 7 YDKSLSVQPE 0.000 47 VHQKLVNDPR 0.006 83 TSASEEEKYD 0.000 34 LKEEQGRAFR 0.006 77 SFR SETSAS 0.000 46 SVHQKLVNDP 0.006 66 VGDIVGRDLS 0.000 67 GDIVGRDLSI 0.005 1 MGKCQEYDKS · 0,000 76 ISFRNSETSA 0.005 21 LRDENGECGQ 0.000 QTFRLKEEQG- 0.005 22 RDENGECGQT 0.000 57 ETQEVFGGGV 0.004 78 FRNSETSASE 0.000 68 DIVGRDLSIS 0.004 55 PRETQEVFGG 0.000 . 75 SISFRNSETS 0.004 72 RDLSISFRNS 0.000 31 TFRLKEEQGR 0.004 36 EEQGRAFRGS 0.000 24 ENGECGQTFR 0.004 28 CGQTFRLKEE 0.000 23 DENGECGQTF 0.003 11 LSVQPEKRTG 0.000 2 GKCQEYDKSL 0.003 49 QKLVNDPRET 0.000 51 LVNDPRETQE 0.002 6 EYDKSLSVQP 0.000 53 NDPRETQEVF 0.002 16 EKRTGLRDEN 0.000 NGECGQTFRL 0.002 15 PEKRTGLRDE 0.000 29 GQTFRLKEEQ 0.002 3 KCQEYDKSLS 0.002 40 RAFRGSSVHQ 0.001 18 RTGLRDENGE 0.001 42 FRGSSVHQKL 0.001 74 LSISFRNSET 0.001 44 GSSVHQKLVN 0,001 39 GRAFRGSSVH 0.001 48 HQKLV DPRE 0.001 52 V DPRETQEV 0.001 KEEQGRAFRG 0.001 86 SEEEKYDMSG 0.001 61 VFGGGVGDIV 0.000 QEYDKSLSVQ 0.000 156 Table X v.5-A3-10iners: 213P1F11 157 Table XI v. l-Al l-9mers: 213P1F11 Table XI v. l-Al l-9mers: 213P1F11 Pos 1234 56789 Score Pos 123456789 Score 189 QTLVDVFTK 4.500 Portion of 141 ETVGGDEIV 0.005 Portion of 125 KVYI IQACR 2.400 SEQ ID 94 FLKGEDGEM . 0.004 SEQ ID KARK 2.000 NO: 3; 218 LVQEG 80 SCAFWLMA 0.004 NO: 3; each stall each start 191 LVDVFTKRK 1.000 position is 111 ALNNKNCQA 0.004 position is 107 NLFEALNNK 0.800 specified, 102 MVKLENLFE 0.004 specified, 59 EQFQEELEK 0.720 the length 86 LMAHGREGF 0.004 the length 83 FWLMAHGR 0.600 of each 148 IVMVI KDS P 0.004 of each 171 TVEGYIAYR 0.400 peptide is 9 160 PTYTDALHV ' 0.004 peptide is 9 amino 45 QLRFESTMK 0.400 187 amino FIQTLVDVF acids, the 0.004 acids, the 175 YIAYRHDQK 0.400 end 151 . VI KDS PQTI 0.004 end 19 LALI LCVTK 0.300 position for 161 TYTDALHVY 0.004 position for 117 CQALRAKPK 0.300 each 71 AI DSREDPV 0.004 each 3 NPRS LEEEK 0.200 peptide is 168 VYSTVEGYI 0.004 peptide is 96 KGEDGEMVK 0.120 the start 158 TI PTYTDAL 0.004 the start position position 129 IQACRGEQR 0.120 plus eight' 100 GEMVKLENL 0.004 plus eight 190 TLVDVFTKR 0.120 226 KTNPEIQST 0.003 67 KFQQAI DSR 0.120 150 MVI KDS PQT 0.003 88 AHGREGFLK 0.060 118 QALRAKPKV ' 0.003 145 GDEIVMVI K 0.060 186 CFIQTLVDV 0.003 215 EAELVQEGK 0.060 84 WLMAHGRE 0.003 21 LI LCVTKAR 0.060 231 IQSTLRKRL 0.003 142 TVGGDEIVM 0.040 77 DPVSCAFW 0.003 228 NPEI QSTLR 0.040 212 RMAEAELVQ 0.002 167 HVYSTVEGY 0.040 13 DMS GARLAL 0.002 204 ELLT EVTRR . 0.036 10 EKYDMS GAR 0.002 233 STLRKRLYL 0.030 230 EIQSTLRKR 0.002 170 STVEGYIAY 0.030 57 TAEQFQEEL 0.002 44 RQLRFESTM 0.027 159 I PTYTDALH 0.002 50 STMKRDPTA 0.020 60 QFQEELEKF 0.002 217 ELVQEGKAR 0.018 119 ALRAKPKVY 0.002 229 PEIQSTLRK 0.018 87 MAHGREGFL 0.002 203 LELLTEVTR 0.018 194 VFT RKGHI 0.002 46 LRFESTMKR 0.016 78 PVSCAFWL 0.002 157 QTI PTYTDA 0.015 24 CVTKAREGS 0.002 115 KNCQALRAK 0.012 174 GYIAYRHDQ 0.002 121 RAKPKVYI I 0.012 135 EQRDPGETV 0.002 39 LEHMFRQLR 0.012 75 REDPVSCAF 0.002 104 KLENLFEAL 0.012 101 EMVKLENLF 0.002 123 KPKVYI IQA ' 0.012 140 GETVGGDEI' 0.002 1 1 KYDMSGARL 0.012 97 GEDGEMVKL 0.002 195 FTKRKGHI L 0.010 16 GARLALI LC 0.001 36 LDALEHMFR 0.008 21 1 RRMAEAELV 0.001 81 CAFWLMAH 0.008 197 KRKGHI LEL 0:001 113 NNKNCQALR 0.008 144 GGDEIVMVI 0.001 6 SLEEEKYDM 0.008 18 RLALI LCVT 0.001 201 HI LELLTEV 0.006 64 ELEKFQQAI 0.001 193 DVFTKRKGH 0.006 172 VEGYIAYRH 0.001 ALI LCVTKA 0.006 35 DLDALEHMF 0.001 223 KARKTNPEI 0.006 162 YTDALHVYS 0.001 208 EVTRRMAEA 0.006 206 LTEVTRRMA 0.001 158 Table XI v.2-Al l-9mers Table XI v.2 -Al l - 9mers Pos 123456789 Score YSTVEGPTP 0.000 Portion of | 28 LWNSQDTS P 0.000 SEQ ID EGPTPFQDP 0.000 NO: 5; each stail 24 PNP PLWNSQ 0.000 position is 19 PSEAPPNPP 0.000 specified, 49 PWWMCSRRG 0.000 the length of each 1 peptide is 9| amino acids, the end J position for| each peptide is the start position plus eight 159 Table XI v.3-All-9mers 160 Table XI v.4-All-9mers: 213P1F11 Table XI v.4-All-9mers: 213P1F11 Pos 123456789 Score Pos 123456789 Score 82 ETSASEEEK 0.300 Portion of 79 RNSETSASE 0.000 Portion of | 48 HQKLVNDPR 0.120 SEQED 63 GGGVGDIVG 0.000 SEQ ID KSLSVQPEK 0.090 NO: 9; 6 EYDKSLSVQ 0.000 NO: 9; 9 each start each start SLSVQPEKR 0.080 position is 24 ENGECGQTF 0.000 position is 1 MGKCQEYDK 0.040 specified, 84 SASEEEKYD 0.000 specified, 14 QPEKRTGLR 0.040 the length 67 GDIVGRDLS 0.000 the length KEEQGRAFR 0.036 of.each 59 QEVFGGGVG 0.000 of each 42 FRGSSVHQK 0.020. peptide is 9 17 KRTGLRDEN 0.000 peptide is 9| amino amino 64 GGVGDIVGR 0.018 45 SSVHQKLV 0.000 acids, the acids, the 71 GRDLSISFR 0.012 end 22 RDENGECGQ 0.000 end 33 RLKEEQGRA 0.012 position for 2, GKCQEYDKS 0.000 position for| 40 RAFRGSSVH 0.012 each 86 SEEEKYDMS 0.000 each 32 FRLKEEQGR 0.006 peptide is 81 SETSASEEE 0.000 peptide is 0.006 the start 36 EEQGRAFRG 0.000 the start 65 GVGDIVGRD position position 27 ECGQTFRLK 0.006 plus eight 52 VNDPRETQE 0.000 ' plus eight 58 TQEVFGGGV 0.006 28 CGQTFRLKE 0.000 13 VQPEKRTGL 0.006 76 ISFRNSETS ■ 0.000 26 GECGQTFRL 0.005 74 LSISFRNSE 0.000 NGECGQTFR 0.004 49 QKLVNDPRE 0.000 68 DIVGRDLSI 0.004 19 TGLRDENGE 0.000 3 KCQEYDKSL 0.003 47 ■ VHQKLVNDP ο.οοο· 18 RTGLRDENG 0.003 66 VGDIVGRDL 0.000 QEYDKSLSV 0.002 38 QGRAFRGSS 0.000 77 SFRNSETSA 0.002 34 LKEEQGRAF 0.000 46 SVHQKLVND 0.002 80 NSETSASEE 0.000 51 LV DPRETQ 0.002 7 YDKSLSVQP 0.000 QTFRLKEEQ 0.002 78 FRNSETSAS 0.000 69 IVGRDLSIS. 0.002 21 LRDENGECG 0.000 61 VFGGGVGDI 0.002 37 EQGRAFRGS 0.000 GLRDENGEC 0.001 23 DENGECGQT 0.000 60 EVFGGGVGD 0.001 11 LSVQPEKRT 0.000 29 GQTFRLKEE 0.001 15 PEKRTGLRD 0.000 43 RGSSVHQKL 0.001 73 DLSISFRNS 0.000 4 CQEYDKSLS 0.001 8 DKSLSVQPE 0.000 39 GRAFRGSSV 0.001 55 PRETQEVFG 0.000 54 DPRETQEVF 0.001 16 EKRTGLRDE 0.000 56 RETQEVFGG 0.001 75 SISFRNSET 0.000 85 ASEEEKYDM 0.000 70 VGRDLSISF 0.000 44 GSSVHQKLV 0.000 57 ETQEVFGGG 0.000 72 RDLSISFRN 0.000 41 AFRGSSVHQ 0.000 53 NDPRETQEV 0.000 31 TFRLKEEQG 0.000 83 TSASEEEKY 0.000 62 FGGGVGDIV 0.000 12 SVQPEKRTG 0.000 50 KLVNDPRET 0.000 161 Table XI v.5-All-9mers: 213P1F11 Pos 123456789 Score 4 LALILRVTK 0.300 Portion of 1 GARLALILR 0.240 SEQID LILRVTKAR 0.060 NO: 11, 6 each stiirt 9 RVTKAREGS 0.006 position is ALILRVTKA 0.006 specified, 2 ARLALILRV 0.001 the length 7 ILRVTKARE 0.000 of each 3 RLALILRVT 0.000 peptide is 9 amino 8 LRVTKAREG 0.000 acids, the end position for each peptide is the start position phis eight Table XI V.6-A1 l-9mers: 213P1F11 162 Table XII v.1-Al 1-lOniers: 213P1F11 Table XII v.l-All-10mers: 213P1F11 Portion of SEQ ID NO: 3; each start position is specified, the length of eachj peptide is 10 amino acids, the end position for each peptide is the start position plus nine 163 141 ETVGGDEIVM 0.009 56 PTAEQFQEEL 0.001 168 VYSTVEGYIA 0.008 25 VTKAREGSEE 0.001 112 LNNKNCQALR 0.008 200 GHILELLTEV 0.001 85 VLMAHGREGF 0.008 207 TEVTRRMAEA 0.001 227 TNPEIQSTLR 0.008 77 DPVSCAFWL 0.001 27 KAREGSEEDL 0.006 110 EALNNKNCQA 0.001 117 CQALRAKPKV 0.006 12 YDMSGARLAL 0.001 1 1 1 ALNNKNCQAL 0.004 107 NLFEALNNKN 0.001 80 SCAFWLMAH 0.004 1 13 NNKNCQALRA 0.001 185 SCFIQTLVDV 0.004 123 KPKVYIIQAC 0.001 94 FLKGEDGEMV 0.004 179 RHDQKGSCFI 0.001 86 LMAHGREGFL 0.004 129 IQACRGEQRD 0.001 164 Table XII V.2-A11-lOmers: 213P1F11 Table XII V.2-A11-lOmers: 213P1F1 1 Pos 1234567890 Score 24 PPNPPLWNSQ 0.000 Portion of | 29 LWNSQDTS PT 0.000 SEQ ID 55 SRRGKDI SWN 0.000 NO: 5; each stall TS PTDMI RKA 0.000 position is | PS EAP PNP PL 0.000 specified, PNPPLWNSQD 0.000 the length | of each peptide is 10 amino acids, the end Jposition for] each peptide is the start position plus nine 165 Table XII v.3-All-10mers: 213P1F11 166 Table XII v.4-All-10mers: 213P1F11 Table XII V.4-A11 -1 Omers: 213P1F11 Pos 1234567890 Score 56 RETQEVFGGG 0.000 68 DIVGRDLSIS 0.000 23 DENGECGQTF 0.000 37 EQGRAFRGSS 0.000 79 RNSETSASEE 0.000 44 GSSVHQKLVN 0.000 EYDKSLSVQP 0.000 73 DLSISFRNSE 0.000 QEYDKSLSVQ 0.000 86 SEEEKYDMSG 0.000 27 ECGQTFRLKE 0.000 64 GGVGDIVGRD 0.000 59 QEVFGGGVGD 0.000 17 KRTGLRDENG 0.000 22 RDENGECGQT 0.000 54 DPRETQEVFG 0.000 62 FGGGVGDIVG 0.000 45 SSVHQKLVND 0.000 19 TGLRDENGEC 0.000 74 LSISFRNSET 0.000 80 NSETSASEEE 0.000 MGKCQEYDKS 0.000 21 LRDENGECGQ 0.000 85 ASEEEKYDMS 0.000 66 VGDIVGRDLS 0.000 YDKSLSVQPE 0.000 78 FRNSETSASE 0.000 83 TSASEEEKYD 0.000 28 CGQTFRLKEE 0.000 72 RDLSISFRNS 0.000 55 PRETQEVFGG 0.000 16 EKRTGLRDEN 0.000 11 LSVQPEKRTG 0.000 49 QKLVNDPRET 0.000 36 EEQGRAFRGS 0.000 PEKRTGLRDE 0.000 167 168 Table XIII v. l-A24-9mers: 213P1F11 Table XIII v. l-A24-9mers: 213P1F11 Pos 1234 567 8 9 Score Pos 1234567 8 9 Score 1 1 KYDMSGARL 400.000 Portion of 226 KTN PEIQST 0.432 Portion of 168 VYSTVEGYI 70.000 SEQ ED 179 RHDQKGSCF 0.400 SEQ ID NO: 3, 60 QFQEELEKF 19.800 40 EHMFRQLRF 0.300 NO: 3; each stall each start 104 KLENLFEAL 17.280 position is 18 RLALI LCVT 0.280 position is 227 TNPEIQSTL 10.080 · specified, 199 KGHI LELLT 0.240 specified, 183 KGSCFIQTL 9.600 the length 201 HI LELLTEV 0.238 the length 112 LNNKNCQAL ,7.200 of each 99 DGEMVKLEN 0.231 of each 31 GSEEDLDAL 7.200 peptide is 9 155 S PQTI PTYT 0.210 peptide is 9 amino amino 38 ALEHMFRQL 7.200 147 EIVMVI KDS acids, the . 0.210 . acids, the 57 TAEQFQEEL 6.600 end 164 DALHVYSTV 0.210 end 233 STLRKRLYL 6.000 position for 123 KPKVYI IQA 0.200 position for 158 TI PTYTDAL 6.000 each 157 QTI PTYTDA 0.180 each 161 TYTDALHVY 6.000 peptide is 202 I LELLTEVT 0.180 peptide is AYRHDQKGS 5.000 the start 177 170 STVEGYIAY 0.180 the start position position 194 VFTKRKGHI 5.000 plus eight 20 ALI LCVTKA 0.165 plus eight SGARLALI L 4.800 219 VQEGKARKT 0.165 23 1 IQSTLRKRL 4.800 118 QALRAKPKV 0.165 101 EMVKLENLF 4.320 50 STMKRDPTA 0.150 87 MAHGREGFL 4.000 150 MVI KDS PQT 0.150 195 FTKRKGHI L 4.000 47 RFESTMKRD 0. 150 13 DMSGARLAL 4.000 154 DS PQTI PTY 0.150 187 FIQTLVDVF 3.600 206 LTEVTRRMA 0.150 . DLDALEHMF 2.400 106 ENLFEALNN 0.150 121 RAKPKVYI I 2.400 111 ALNNKNCQA 0.150 223 KARKTNPEI 2.200 67 KFQQAI DSR 0.150 86 LMAHGREGF 2.000 141 ETVGGDEIV 0.150 64 ELEKFQQAI 1.800 77 DPVSCAFW 0.150 144 GGDEIVMVI 1.680 180 HDQKGSCFI 0.150 44 RQLRFESTM 1.500 80 SCAFWLMA 0.140 151 VI KDS PQTI 1.440 188 IQTLVDVFT 0.140 108 LFEALNNKN 1.188 184 GSCFIQTLV 0.140 198 RKGHI LELL 1.120 30 EGS EEDLDA 0.120 14 MSGARLALI 1.000 143 VGGDEIVMV 0.120 6 SLEEEKYDM 0.900 162 YTDALHVYS . 0.120 197 KRKGHI LEL 0.880 73 DS REDPVSC 0.120 205 LLTEVTRRM 0.840 135 EQRDPGETV 0.120 126 VYI IQACRG 0.750 208 EVTRRMAEA 0.1 10 174 GYIAYRHDQ 0.750 140 GETVGGDEI 0.1 10 186 CFIQTLVDV 0.750 232 QSTLRKRLY 0.100 75 REDPVSCAF 0.672 16 GARLALI LC 0.100 42 MFRQLRFES 0.660 119 ALRAKPKVY 0.100 100 GEMVKLENL 0.600 167 HVYSTVEGY 0.100 28 AREGS EEDL 0.600 24 CVTKAREGS 0.100 94 , FLKGEDGEM 0.550 49 ESTMKRDPT 0.100 79 VSCAFWLM 0.500 71 AI DS REDPV 0.100 142 TVGGDEIVM 0.500 120 LRAKPKVYI 0.100 78 PVSCAFWL 0.480 169 YSTVEGYIA 0.100 53 KRDPTAEQF ' 0.480 34 EDLDALEHM 0.090 ' 97 GEDGEMVKL " 0.440 82 AFWLMAHG 0.090 210 TRRMAEAEL 0.440 93 GFLKGEDGE 0.075 Table XIII v.2-A24-9mers: 213P1F11 Table XIII v.2-A24-9mers: 213P1F1 1 Pos 123456789 Score 13 QDPLYLPSE 0.002 Portion of | 36 PTDMI RKAH 0.001 SEQ ID VEGPTP FQD 0.001 NO: 5; each start 40 I RKAHALSR 0.001 position is 49 PW MCS RRG 0.001 specified, PLYLPS EAP 0.001 the length of each ] peptide is 9| amino acids, the end J position for| each peptide is the stmt position plus eight 170 Table XIII v.3-A24-9mers: 213P1F11 171 Table ΧΠΙ v.4-A24-9mers: 213P1F11 Table XIII v.4-A24-9iners: 213P1F11 Pos 123456789 Score 64 GGVGDIVGR 0.015 Portion of 67 GDIVGRDLS 0.015 SEQID 78 FRNSETSAS 0.015 NO: 9; each start 14 QPEKRTGLR 0.015 position is 48 HQKLVNDPR 0.014 specified, QTFRLKEEQ 0.013 the length QEYDKSLSV 0.012 of each 52 V DPRETQE 0.012 I peptide is 9| amino 27 ECGQTFRLK 0.012 acids, the 84 SASEEEKYD 0.012 end GKCQEYDKS 0.011 J position for| SLSVQPEKR 0.011 each 29 GQTFRLKEE 0.011 peptide is 82 ETSASEEEK 0.011 the start position 39 GRAFRGSSV 0.010 plus eight 46 SVHQKLVND 0.010 60 EVFGGGVGD 0.010 63 GGGVGDIVG 0.010 MGKCQEYDK 0.010 KEEQGRAFR 0.003 22 RDENGECGQ 0.003 47 VHQKLVNDP 0.002 56 RETQEVFGG 0.002 32 FRLKEEQGR 0.002 59 QEVFGGGVG 0.002 36 EEQGRAFRG 0.002 49 QKLV DPRE 0.002 DKSLSVQPE 0.001 YDKSLSVQP 0.001 21 LRDENGECG 0.001 81 SETSASEEE 0.001 42 FRGSSVHQK 0.001 16 EKRTGLRDE 0.001 71 GRDLSISFR 0.001 55 PRETQEVFG 0.000 PEKRTGLRD 0.000 172 Table XIII v.5-A24-9mers: 213P1F11 Table XIII v.6-A24-9mers: 213P1F11 173 Table XIV v. l-A24-10mers: 213P1F11 Table XIV v. l-A24-10mers: 213PIF11 Pos 1234567890 Score Pos 1234567890 Score 226 KTNPEIQSTL 20.160 Portion of 52 MKRDPTAEQF 0.240 Portion of 194 VFTKRKGHI L 20.000 SEQ ID 178 YRHDQKGSCF 0.240 SEQ ID CFIQTLVDVF 18.000 NO: 3; 186 63 EELEKFQQAI 0.216 NO: 3; each start each start 96 KGEDGEMVKL 15.840 position is 201 HI LELLTEVT 0.216 position is 11 KYDMSGARLA 10.000 specified, 67 KFQQAI DSRE 0.210 specified, 27 KAREGSEEDL ' 9.600 the length 187 FIQTLVDVFT 0.210 the length 37 DALEHMFRQL ' 8.640 of each 154 DS PQTI PTYT 0.210 of each 161 TYTDALHVYS 7.200 peptide is 179 RHDQKGSCFI 0.200 peptide is amino L 10 amino 111 ALNNKNCQA 7.200 acids, the 39 LEHMFRQLRF 0.200 acids, the 230 EIQSTLRKRL 7.200 end 218 LVQEGKARKT 0.198 end 77 DPVSCAFWL 7.200 position for 107 NLFEALNNKN 0. 190 position for 157 QTI PTYTDAL 7.200 each 6 SLEEEKYDMS 0. 180 each 99 DGEMVKLENL 6.000 peptide is 70 QAI DSREDPV 0.180 peptide is the start 177 AYRHDQKGSC 5.000 139 PGETVGGDEI 0.165 the stall position . position 168 VYSTVEGYIA 5.000 plus nine 19 LALI LCVTKA 0.165 plus nine 14 MSGARLALI L 4.800 215 EAELVQEGKA 0.165 EGSEEDLDAL 4.800 47 RFESTMKRDP 0.150 209 VTRRMAEAEL 4.400 23 LCVTKAREGS 0.150 93 GFLKGEDGEM 4.125 1 18 QALRAKPKVY 0.150 232 QSTLRKRLYL 4.000 149 VMVI KDS PQT 0.150 86 LMAHGREGFL 4.000 110 EALNNK CQA 0.150 85 VLMAHGREGF 3.000 219 VQEGKARKTN 0.150 59 EQFQEELEKF 2.200 79 VSCAFWLMA 0.140 150 MVI KDS PQTI 1.800 41 HMFRQLRFES 0.132 RSLEEEKYDM 1.800 73 DSREDPVSCA 0.120 143 VGGDEIVMVI 1.680 181 DQKGSCFIQT 0.120 167 HVYSTVEGYI 1.400 205 LLTEVTRRMA 0.120 197 KRKGHI LELL 1.120 16 GARLALI LCV 0.120 204 ELLTEVTRRM 1.050 102 MVKLENLFEA 0.1 10 103 VKLENLFEAL 1.037 3 NPRSLEEEKY 0. 1 10 1 19 ALRAKPKVYI 1.000 222 GKARKTNPEI 0.110 193 DVFTKRKGHI 1.000 1 17 CQALRAKPKV 0.110 13 DMSGARLALI 1.000 60 QFQEELEKFQ 0.108 141 ETVGGDEIVM 0.750 169 YSTVEGYIAY 0.100 108 LFEALNNKNC 0.750 176 IAYRHDQKGS 0.100 126 VYIIQACRGE 0.750 185 SCFIQTLVDV 0.100 174 GYIAYRHDQK 0.750 120 LRAKPKVYI I 0.100 12 YDMSGARLAL 0.600 231 IQSTLRKRLY 0.100 42 MFRQLRFEST 0.600 113 NNKNCQALRA 0.100 56 PTAEQFQEEL 0.528 142 TVGGDEIVMV 0.100 74 SREDPVSCAF 0.504 94 FLKGEDGEMV 0.100 EKYDMSGARL 0.480 49 . ESTMKRDPTA 0.100 182 QKGSCFIQTL 0.480 15 SGARLALI LC 0.100 196 TKRKGHI LEL 0.440 162 YTDALHVYST 0.100 100 GEMVKLENLF 0.432 71 AI DSREDPVS 0.100 34 EDLDALEHMF 0.432 159 I PTYTDALHV 0.100 123 KPKVYI IQAC 0.336 82 AFWLMAHGR 0.075 . 133 RGEQRDPGET 0.330 43 FRQLRFESTM 0.075 104 KLENLFEALN ' 0.300 78 PVSCAFWLM 0.050 183 KGSCFIQTLV 0.280 33 EEDLDALEHM 0.050 Table XIV v.2-A24-10mers: 213P1F11 Table XIV v.2-A24-10mers: 213P1F11 Pos 1234567890 Score LHVYSTVEGP 0.002 50 PWWMCSRRGK 0.001 28 PLWNSQDTSP 0.001 33 QDTSPTDMIR 0.001 41 IRKAHALSRP 0.001 16 PLYLPSEAPP 0.001 175 Table XIV v.3-A24-10mers: 213P1F11 ) 176 Table XIV v.4-A24-10mers: 213P1F11 Table XIV V.4-A24-1 Oiners: 213P1F11 Pos 1234567890 Score 11 LSVQPEKRTG 0.015 14 QPEKRTGLRD 0.015 32 FRLKEEQGRA 0.015 45 SSVHQKLVND 0.015 46 SVHQKLVNDP 0.014 71 GRDLSISFRN 0.014 16 EKRTGLRDEN 0.013 29 GQTFRLKEEQ 0.013 70 VGRDLSISFR 0.012 73 DLSISFRNSE 0.012 GLRDENGECG 0.012 54 DPRETQEVFG 0.012 24 ENGECGQTFR 0.012 27 ECGQTFRLKE 0.011 QTFRLKEEQG 0.010 83 TSASEEEKYD 0.010 63 GGGVGDIVGR 0.010 48 HQKLVNDPRE 0.010· 62 FGGGVGDIVG 0.010 56 RETQEVFGGG 0.003 KEEQGRAFRG 0.003 47 VHQKLWDPR 0.002 17 KRTGLRDENG 0.002 34 LKEEQGRAFR 0.002 86 SEEEKYDMSG 0.002 DKSLSVQPEK 0.002 59 QEVFGGGVGD 0.002 78 FRNSETSASE 0.002 26 GECGQTFRLK 0.001 YDKSLSVQPE 0.001 21 LRDENGECGQ 0.001 QEYDKSLSVQ 0.001 SETSASEEEK 0.001 39 GRAFRGSSVH 0.001 55 PRETQEVFGG 0.000 PEKRTGLRDE 0.000 177 Table XIV v.5-A24-10mers: 213P1F11 Table XIV v.6-A24-I0mers: 213P1F11 Pos 1234567890 Score 9 AMNNKNCQAL 7.200 Portion of 6 LFEAMNNKNC 0.750 SEQ ID 2 KLENLFEAMN 0.300 NO: 13; each start NLFEAMNNKN 0.158 position is 8 EAMNNKNCQA . 0.150 specified, 1 VKLENLFEAM 0.130 the length 4 ENLFEAMNNK■ 0.018 of each 3: LENLFEAMNN 0.015 peptide is amino M NKNCQALR 0.015 acids, .the 7 FEAMNNKNCQ 0.001 end position for each peptide is the start position plus nine Table XV v. l -B7-9mers: 213P1F11 Table XV v. l-B7-9mers: 213P1F11 Pos 123456789 Score Pos 123456789 Score 87 MAHGREGFL 12.000 Portion of 143 VGGDEIVMV 0.200 Portion of 223 KARKTNPEI 12.000 SEQ ID 201 HI LELLTEV 0.200 SEQ ID NO: 3; 13 DMSGARLAL 6.000 138 . DPGETVGGD 0.200 NO: 3; each start each start 233 STLRKRLYL 6.000 position is 141 ETVGGDEIV 0.200 position is 23 1 IQSTLRKRL 6.000 specified, 71 AI DS REDPV 0.180 specified, 142 TVGGDEIVM 5.000 the length 24 CVTKAREGS 0.150 the length 1 12 LNNKNCQAL 4.000 of each 148 IVMVI KDS P 0.150 of each 227 TNPEIQSTL 4.000 peptide is 9 49 ESTMKRDPT 0. 150 peptide is 9 amino TRRMAEAEL amino 210 4.000 97 GEDGEMVKL acids, the 0.120 acids, the 183 . KGSCFIQTL 4.000 end 144 GGDEIVMVI 0.120 end 77 DPVSCAFW 4.000 position for 11 KYDMSGARL 0.120 position for 158 TI PTYTDAL 4.000 each 64 ELEKFQQAI 0.120 each SGARLALI L 4.000 peptide is 199 KGHI LELLT 0.100 peptide is 195 FTKRKGHIL 4.000 the start 80 S CAFWLMA ' 0.100 the start position position 57 TAEQFQEEL 3.600 plus eight 34 EDLDALEHM 0.100 plus eight 38 ALEHMFRQL 3.600 89 HGREGFLKG 0.100 . 16 GARLALI LC 3.000 234 TLRKRLYLQ 0.100 135 EQRDPGETV 3.000 45 QLRFESTMK 0.100 3 NPRSLEEEK 2.000 188 IQTLVDVFT 0.100 155 S PQTI PTYT 2.000 167 HVYSTVEGY 0.100 78 PVSCAFWL 2.000 18 RLALI LCVT 0.100 123 KPKVYI IQA 2.000 226 KTNPEIQST 0.100 31 GSEEDLDAL 1.200 209 VTRRMAEAE 0.100 121 RAKPKVYI.I 1 .200 157 QTI PTYTDA 0.100 100 GEMVKLENL 1.200 169 YSTVEGYIA 0.100 104 KLENLFEAL 1.200 30 EGSEEDLDA 0.100 94 FLKGEDGEM 1.000 193 DVFTKRKGH 0.075 73 DSREDPVSC 1.000 177 AYRHDQKGS 0.060 79 VSCAFWLM 1.000 120 LRAKPKVYI 0.060 44 RQLRFESTM 1.000 211 RRMAEAELV 0.060 205 LLTEVTRRM 1.000 17 ARLALI LCV 0.060 118 QALRAKPKV 0.600 228 NPEIQSTLR 0.060 164 ■ DALHVYSTV 0.600 218 LVQEGKARK 0.050 119 ALRAKPKVY 0.600 125 KVYI IQACR 0.050 208 EV RRMAEA 0.500 102 MVKLENLFE 0.050 150 MVI KDS PQT 0.500 84 WLMAHGRE 0.050 197 KRKGHI LEL 0.400 83 FWLMAHGR 0.050 198 RKGHI LELL 0.400 . 70 QAI DS REDP 0.045 14 MSGARLALI 0.400 206 LTEVTRRMA 0.045 151 VI KDS PQTI 0.400 140 GETVGGDEI 0.040 28 AREGSEEDL 0.360 168 VYSTVEGYI 0.040 131 ACRGEQRDP 0.300 194 VFTKRKGHI 0.040 27 KAREGS EED 0.300 180 HDQKGSCFI 0.040 111 ALNNKNCQA 0.300 86 LMAHGREGF 0.030 ALI LCVTKA 0.300 19 LALI LCVTK 0.030 6 SLEEEKYDM 0.300 37 DALEHMFRQ 0.030 50 STMKRDPTA 0.300 12 YDMSGARLA 0.030 55 DPTAEQFQE 0.200 165 ALHVYSTVE 0.030 184 GSCFIQTLV 0.200 85 VLMAHGREG 0.030 159 I PTYTDALH 0.200 216 AELVQEGKA 0.030 Table XV v.2-B7-9mers Table XV V.2-B7- 9mers Pos 123456789 Score Pos 123456789 Score 8 GPTPFQDPL 80.000 Portion of 16 LYLPSEAPP 0.001 Portion of TPFQDPLYL 80.000 SEQID 28 LWNSQDTSP 0.001 SEQID AHAL 4.000 N 38 DMIRK O: 5; 19 PSEAPPNPP 0.000 NO: 5; each start each start 44 HALSRPWWM 3.000 position is 36 PTDMIRKAH 0.000 position is 14 DPLYLPSEA .2.000 specified, 11 PFQDPLYLP 0.000 specified, SPTDMIRKA 2.000 the length 49 PWWMCSRRG 0.000 the length 22 APPNPPLWN 1.800 of each of each SEAPPNPPL 0.600 peptide is 9 peptide is 9 amino amino 45 ALSRPWWMC 0.450 acids, the acids, the 52 MCSRRGKDI 0.400 end end 31 SQDTSPTDM 0.300 position for position for 48 RPWWMCS R 0.200 each each 46 LSRPWWMCS 0.200 peptide is peptide is the start the start 53 CSRRGKDIS 0.200 position position 39 MIRKAHALS 0.200 plus eight plus eight NPPLWNSQD 0.200 26 PP-LWNSQDT 0.200 18 LPSEAPPNP 0.200 29 WNSQDTSPT 0.100 42 KAHALSRPW 0.060 32 QDTSPTDMI 0.060 23 PPNPPLWNS 0.060 21 EAPPNPPLW 0.060 1 HVYSTVEGP 0.050 37 TDMIRKAHA 0.030 TVEGPTPFQ 0.023 54 SRRGKDISW 0.020 56 RGKDISWNF 0.020 4' STVEGPTPF 0.020 17 YLPSEAPPN 0.020 7 EGPTPFQDP 0.015 51 WMCSRRGKD 0.015 33 DTSPTDMIR 0.010 2 VYSTVEGPT 0.010 34. TSPTDMIRK 0.010 NSQDTSPTD 0.010 3 YSTVEGPTP 0.010 43 AHALSRPWW 0.009 12 FQDPLYLPS 0.006 50 WWMCSRRGK 0.005 9 PTPFQDPLY 0.002 55 RRGKDIS N 0.002 27 PLWNSQDTS ' 0.002 PLYLPSEAP 0.002 41 RKAHALSRP 0.001 6 VEGPTPFQD 0.001 13 QDPLYLPSE 0.001 40 IRKAHALSR 0.001 47 SRPWWMCSR 0.001 24 PNPPLWNSQ 0.001 Table XV v.3-B7-9mers: 213P1F11 Pos 123456789 Score TLPSPFPYL 6.000 Portion of 3 IQACRGATL 4.000 SEQID ACRGATLPS 0.600 • NO: 7, each stitrt 12 PSPFPYLSL 0.600 position is 11 LPSPFPYLS 0.400 specified, 2 IIQACRGAT 0.150 the length 1 YIIQACRGA 0.100 of each 9 ATLPSPFPY 0.060 peptide is 9 amino 8 GATLPSPFP 0.045 acids, the 4 QACRGATLP 0.030 end 7 RGATLPSPF 0.020 position for 6 CRGATLPSP 0.001 each peptide is the start position plus eight Table XV v.4-B7-9mers: 213P1F11 Table XV v.4-B7-9mers: 213P1F11 Pos 123456789 Score Pos 123456789 Score 13 VQPEKRTGL 6.000 Portion of 64 GGVGDIVGR 0.010 3 KCQEYDKSL 4.000 SEQ ID 18 RTGLRDENG 0.010 QEVF 4.000 NO: 9; 54 DPRET 16 EKRTGLRDE 0.010 each start 43 RGS SVHQKL 4.000 position is 48 HQKLVNDPR 0.010 66 VGDIVGRDL 1.200 specified, 29 GQT FRLKEE 0.010 GLRDENGEC 1.000 the length 79 RNS ETSAS E 0.010 85 ASEEEKYDM 0.900 - of each 19 T.GLRDENGE 0.010 26 GECGQTFRL 0.400 peptide is 9 31 TFRLKEEQG 0.010 amino 68 DIVGRDLS I 0.400 4 CQEYDKSLS 0.006 acids, the 38 QGRAFRGSS 0.300 ■ end 67 GDIVGRDLS 0.003 62 FGGGVGDIV 0.200 position for 17 KRTGLRDEN 0.003 70 VGRDLSI SF 0.200 each 52 VNDPRETQE 0.003 44 GSSVHQKLV 0.200 peptide is 25 NGECGQTFR 0.003 the start 11 LSVQPEKRT 0.150 80 NS ETSASEE 0.003 position 51 LVNDPRETQ 0.113 plus eight 2 GKCQEYDKS 0.002 77 S FRNSETSA 0.100 72 RDLS I S FRN 0.002 69 IVGRDLS I S 0.100 78 FRNSETSAS 0.002 75 S I S FRNSET 0.100 59 QEVFGGGVG 0.001 33 RLKEEQGRA 0.100 47 VHQKLVNDP 0.001 50 KLVNDPRET 0.100 42 FRGSSVHQK 0.001 14 QPEKRTGLR 0.060 81 SETSAS EEE 0.001 58 TQEVFGGGV 0.060 49 QKLVNDPRE 0.001 60 EVFGGGVGD ■ 0.050 36 EEQGRAFRG 0.001 12 SVQPEKRTG 0.050 8 DKSLSVQPE 0.001 46 SVHQKLVND 0.050 32 FRLKEEQGR 0.001 65 GVGDIVGRD 0.050 7 YDKSLSVQP 0.001 61 VFGGGVGDI 0.040 56 RETQEVFGG 0.001 . 84 SASEEEKYD 0.030 34 LKEEQGRAF 0.001 41 AFRGSSVHQ 0.030 86 SEEEKYDMS 0.001 40 RAFRGSSVH 0.030 35 KEEQGRAFR 0.000 83 TSASEEEKY 0.020 6 EYDKSLSVQ 0.000 37 EQGRAFRGS 0.020 21 LRDENGECG 0.000 73 DLS I S FRNS 0.020 71 GRDLS I S FR 0.000 24 ENGECGQTF 0.020 22 RDENGECGQ 0.000 39 GRAFRGSSV 0.020 15 PEKRTGLRD 0.000 53 NDPRETQEV 0.020 55 PRETQEVFG 0.000 QEYDKSLSV 0.020 45 SSVHQKLVN 0.020 76 I S FRNS ETS 0.020 74 LS I S FRNSE 0.015 28 , CGQTFRLKE 0.015 27 ECGQTFRLK 0.010 9 KSLSVQPE 0.010 23 DENGECGQT 0.010 QTFRLKEEQ 0.010 SLSVQPEKR 0.010 1 MGKCQEYDK 0.010 63 GGGVGDIVG 0.010 82 ETSASEEEK 0.010 57 ETQEVFGGG 0.010 182 Table XV v.5-B7-9mers: 213P1F11 Table XV v.6-B7-9mers: 213P1F11 183 Table XVI v. l -B7-10mers: 213P1F1 1 Table XVI v. l-B7-10mers: 213P1F11 Pos 1234567 8 90 Score Pos 12 34 567 890 Score 27 KAREGSEEDL 120.000 Portion of 138 DPGETVGGDE 0.200 Portion of 77 DPVSCAFWL 80.000 SEQID 55 DPTAEQFQEE 0.200 SEQID TRRMAEAEL 40.000 NO: 3; 210 TRRMAEAELV 0.200 NO: 3, 209 V each start each start 1 19 ALRAKPKVYI 18.000 position is 117 CQALRAKPKV 0.200 position is 11 1 ALNNKNCQAL 12.000 specified, 205 LLTEVTRRMA 0.150 specified, 37 DALEHMFRQL 12.000 the length 148 IVMVIKDSPQ 0.150 the length 16 GARLALILCV 6.000 of each 135 EQRDPGETVG 0.100 of each 232 QSTLRKRLYL 6.000 peptide is 42 MFRQLRFEST 0.100 peptide is amino LRKRL 10 amino 230 EIQST 6.000 79 VSCAFWLMA 0.100 acids, the acids, the 157 QTIPTYTDAL 4.000 end 89 ' HGREGFLKGE 0.100 end 3 NPRSLEEEKY 4.000 position for 113 NNKNCQALRA 0.100 position for 226 KTNPEIQSTL 4.000 each 149 VMVIKDSPQT 0.100 each 196 TKRKGHILEL 4.000 . peptide is 181 DQKGSCFIQT 0. 100 peptide is I PTYTDALHV . the start the start 159 4.000 93 GFLKGEDGEM 0. 100 position position EGSEEDLDAL 4.000 plus nine 43 FRQLRFESTM 0. 100 phis nine 14 MSGARLALIL 4.000 15 SGARLALILC 0.100 86 LMAHGREGFL 4.000 45 QLRFESTMKR 0. 100 193 DVFTKRKGHI 2.000 187 FIQTLVDVFT 0.100 167 HVYSTVEGYI 2.000 201 HILELLTEVT 0. 100 150 MVIKDSPQTI 2.000 154 DSPQTIPTYT 0.100 123 KPKVYIIQAC 2.000 49 ESTMKRDPTA 0.100 12 YDMSGARLAL 1.800 85 VLMAHGREGF 0.090 73 DSREDPVSCA 1.500 215 EAELVQEGKA 0.090 96 KGEDGEMV L 1.200 228 NPEIQSTLRK 0.060 99 DGEMVKLENL 1.200 176 IAYRHDQKGS 0.060 RSLEEEKYDM 1.000 1 18 QALRAKPKVY 0.060 204 ELLTEVTRRM 1.000 84 WLMAHGREG 0.050 142 TVGGDEIVMV 1.000 125 KVYIIQACRG 0.050 141 ETVGGDEIVM 1.000 208 EVTRRMAEAE 0.050 70 QAIDSREDPV 0.600 24 CVTKAREGSE 0.050 78 PVSCAFWLM 0.500 83 FWLMAHGRE 0.050 102 MVKLENLFEA 0.500 63 EELEKFQQAI 0.040 218 LVQEGKARKT . 0.500 120 LRAKPKVYI I 0.040 131 ACRGEQRDPG 0.450 222 GKARKTNPEI 0.040 EKYDMSGARL 0.400 130 QACRGEQRDP 0.030 13 DMSGARLALI . 0.400 134 GEQRDPGETV 0.030 194 VFTKRKGHIL 0.400 23 LCVTKAREGS 0.030 103 VKLENLFEAL 0.400 165 ALHVYSTVEG 0.030 182 QKGSCFIQTL 0.400 17 ARLALILCVT 0.030 143 VGGDEIVMVI 0.400 20 ALILCVTKAR 0.030 56 PTAEQFQEEL 0.400 122 AKPKVYIIQA 0.030 197 KRKGHILELL 0.400 81 CAFWLMAHG 0.030 223 KARKTNPEIQ 0.300 50 STMKRDPTAE 0.030 19 LALILCVTKA 0.300 164 DALHVYSTVE 0.030 155 SPQTIPTYTD 0.300 41 HMFRQLRFES 0.030 177 AYRHDQKGSC 0.300 87 MAHGREGFLK 0.030 110 EALNNKNCQA . 0.300 121 RAKPKVYIIQ 0.030 183 KGSCFIQTLV 0.200 33 . EEDLDALEHM 0.030 94 FLKGEDGE V 0.200 162 YTDALHVYST 0.030 185 SCFIQTLVDV 0.200 133 RGEQRDPGET 0.030 Table XVI- v.2-B7-10mers: 213P1F11 Table XVI- v.2-B7-10mers: 213P1F1 1 Pos 1234567890 Score VYSTVEGPTP 0.001 Portion of | 16 PLYLPSEAP P 0.001 SEQ ID 28 PLWNSQDTS P 0.001 NO: 5; each start LHVYSTVEGP 0.001 position is I 12 P FQDPLYLPS 0.000 specified, 50 PWWMCS RRGK 0.000 the length of each peptide is 10 amino acids, the end position for| each peptide is the start position plus nine 185 Table XVI v.3-B7-10mers: 213P1F11 186 Table XVI v.4-B7-10mers: 213P1F11 Table XVI v.4-B7-10mers: 213P1 F 11 Pos 1234567890 Score 64 GGVGDIVGRD 0.010 29 GQTFRLKEEQ 0.010 48 HQKLVNDPRE 0.010 45 SSVHQKLVND 0.010 32 FRLKEEQGRA 0.010 63 GGGVGDIVGR 0.010 18 RTGLRDENGE 0.010 79 R SETSASEE 0.010 49 QKLVNDPRET 0.010 66 VGDIVGRDLS 0.009 80 NS ETSASEEE 0.003 58 TQEVFGGGVG 0.003 22 RDENGECGQT 0.003 23 DENGECGQTF 0.002 53 NDPRETQEVF 0.002 72 RDLS I S FRNS 0.002 36 EEQGRAFRGS 0.002 '81 SETSASEEEK 0.001 39 GRAFRGS SVH 0.001 YDKSLSVQPE 0.001 26 GECGQTFRLK 0.001 QEYDKSLSVQ 0.001 47 VHQKLA NDPR 0.001 DKSLSVQPEK 0.001 56 RETQEVFGGG 0.001 78 FRNS ETSAS E 0.001 17 KRTGLRDENG 0.001 59 QEVFGGGVGD 0.001 71 GRDLS I S FRN 0.001 34 LKEEQGRAFR 0.000 21 LRDENGECGQ 0.000 KEEQGRAFRG 0.000 EYDKSLSVQP 0.000 86 S EEEKYDMSG 0.000 PEKRTGLRDE 0.000 55 PRETQEVFGG 0.000 187 Table XVI v.6-B7-10mers: 213P1F1 1 188 Table XVII v. l-B35-9mers: 213P1F11 Table XVII v. l -B35-9mers: 213P 1F11 Pos 123456789 Score Pos 123456789 Score 123 KPKVYI IQA 12.000 Portion of 60 QFQEELEKF 0.300 Portion of 232 QSTLRKRLY 10.000 SEQ ID 35 DLDALEHMF 0.300 SEQ ID NO:-3; 7 10.000 AEQFQE 0.300 NO: 3, 9 VSCAFWLM 55 DPT each start each start 154 DS PQTI PTY 10.000 position is 210 RRMAEAEL 0.300 position is 94 FLKGEDGEM 9.000 specified, 5 RS LEEEKYD 0.300 specified, 121 RAKPKVYI I 7.200 the length 226 KTNPEIQST 0.300 the length 223 • KARKTNPEI 7.200 · · of each 38 ALEHMFRQL 0.300 of each 1 19 ALRAKPKVY 6.000 peptide is 9 30 . EGS EEDLDA 0.300 peptide is 9 amino amino 73 DSREDPVSC 4.500 144 ' GGDEIVMVI acids, the 0.240 acids, the 31 GS EEDLDAL 4.500' end 141 ETVGGDEIV 0.200 end 205 LLTEVTRRM 4.000 position for 199 KGHI LELLT 0.200 position for 77 DPVSCAFW 4.000 each 198 RKGHI LELL 0.200 each 170 STVEGYIAY 4.000 peptide is 18 RLALI LCVT 0.200 peptide is the start 44 RQLRFESTM 4.000 159 I PTYTDALH the stall 0.200 position position 195 • FTKRKGHI L 3.000 plus eight 150 MVI KDS PQT 0.150 plus eight 142 TVGGDEIVM 3.000 106 ENLFEALNN 0.150 87 MAHGREGFL 3.000 64 ELEKFQQAI 0.120 151 VI KDS PQTI 2.400 24 CVTKAREGS 0.100 227 TNPEIQSTL ' 2.000 40 EHMFRQLRF 0.100 183 KGSCFIQTL 2.000 1 11 ALNNKNCQA 0.100 167 HVYSTVEGY 2.000 188 IQTLVDVFT 0.100 14 MSGARLALI 2.000 147 EIVMVI KDS 0.100 155 . S PQTI PTYT 2.000 78 PVSCAFWL 0.100 6 SLEEEKYDM 1.800 208 EVTRRMAEA 0.100 135 EQRDPGETV 1.200 100 GEMVKLENL 0. 100 231 IQSTLRKRL 1.000 20 ALI LCVTKA 0.100 233 STLRKRLYL 1.000 157 QTI PTYTDA 0.100 101 E VKLENLF 1.000 80 SCAFWLMA 0.100 158 TI PTYTDAL 1.000 50 STMKRDPTA ' 0.100 1 12 LNNKNCQAL 1.000 89 HGREGFLKG 0.060 187 FIQTLVDVF ■ 1.000 228 NPEIQSTLR 0.060 SGARLALI L 1.000 70 QAI DSREDP 0.060 184 GSCFIQTLV 1.000 95 . LKGEDGEMV 0.060 86 LMAHGREGF 1.000 211 RRMAEAELV 0.060 13 DMSGARLAL 1.000 37 DALEHMFRQ . 0.060 . 16 GARLALI LC 0.900 11 KYDMSGARL 0.060 57 TAEQFQEEL 0.900 71 AI DSREDPV 0.060 169 YSTVEGYIA ,0.750 53 KRDPTAEQF 0.060 118 QALRAKPKV 0.600 179 RHDQKGSCF 0.060 3 NPRSLEEEK 0.600 75 REDPVSCAF 0.060 104 KLENLFEAL- 0.600 1 MSNPRSLEE 0.050 197 KR GHI LEL 0.600 9 EEKYDMSGA 0.045 164 DALHVYSTV 0.600 .177 AYRHDQKGS 0.045 143 VGGDEIVMV 0.600 51 TMKRDPTAE 0.045 49 ESTMKRDPT 0.500 45 QLRFESTMK 0.045 161 TYTDALHVY 0.400 102 ' MVKLENLFE 0.045 138 DPGETVGGD 0.400 13 1 ACRGEQRDP 0.045 34 EDLDALEHM 0.400 97 GEDGEMVKL 0.045 201 HI LELLTEV 0.400 180 HDQKGSCFI 0.040 27 KAREGSEED 0.360 168 VYSTVEGYI 0.040 Table XVII v.2-B35-9mers: 213P1F11 Table XVII v.2-B35-9mers: 213P1F11 Pos 123456789 Score VEGPTPFQD 0.001 Portion of | PLYLPSEAP 0.001 SEQ ID 28 LWNSQDTS P 0.001 NO: 5; each start 36 PTDMI RKAH 0.000 position is 11 PFQDPLYLP 0.000 specified, 49 P WMCSRRG ' 0.000 the length of each peptide is 9| amino acids, the end position for each peptide is the start position plus eight 190 Table XVII v.3-B35-9mers Pos 12-3456789 Score 11 LPSPFPYLS 2.000 Portion of 9 ATLPSPFPY 2.000 SEQ ID RGATLPSPF 2.000 NO: 7; 7 each start TLPSPFPYL 1.000 position is 3 IQAC GATL 1.000 specified, 12 PSPFPYLSL 0.500 the length ACRGATLPS 0.300 of each 1 YIIQACRGA 0.100 peptide is 9 amino 2 IIQACRGAT 0.100 acids, the 4 QACRGATLP . 0.030 end 8 . GATLPSPFP 0.030 position for 6 CRGATLPSP 0.001 each peptide is the start position plus eight Table XVII v.4-B35-9mers Table XVII v.4-B35-9mers Pos 123456789 Score 64 GGVGDIVGR 0.015 78 FRNSETSAS 0.015 GKCQEYDKS 0.015 46 SVHQKLV D 0.010 28 CGQTFRLKE 0.010 29 GQTFRLKEE 0.010 QTFRLKEEQ 0.010 60 EVFGGGVGD 0.010 82 ETSASEEEK 0.010 SLSVQPEKR 0.010 27 ECGQTFRLK 0.010 63 GGGVGDIVG 0.010 67 GDIVGRDLS 0.010 86 SEEEKYDMS 0.006 16 EKRTGLRDE 0.003 YDKSLSVQP 0.003 56 RETQEVFGG 0.003 41 AFRGSSVHQ 0.003 31 TFRLKEEQG 0.003 52 VNDPRETQE 0.003 NGECGQTFR 0.003 32 FRLKEEQGR 0.002 36 EEQGRAFRG 0.001 81 SETSASEEE 0.001 DKSLSVQPE 0.001 49 QKLVNDPRE 0.001 47 VHQKLVNDP 0.001 42 FRGSSVHQK 0.001 59 QEVFGGGVG 0.001 22 RDENGECGQ 0.001 21 LRDENGECG 0.001 KEEQGRAFR 0.001 PEKRTGLRD 0.000 71 GRDLSISFR 0.000 EYDKSLSVQ o.ooo- 55 PRETQEVFG 0.000 192 Table XVII v.5-B35-9mers: 213P1F11 Pos 123456789 . Score 9 RVTKAREGS ' 0.200 Portion of 3 RLALILRVT 0.200 SEQID ALILRV KA 0.100 NO: 11; each start 1 . GARLALILR 0.090 position is 7 ILRVTKARE 0.030 specified, 4 LALILRVTK 0.030 the length 2 ARLALILRV 0.020 of each 6 LILRVTKAR 0.010 peptide is 9 amino 8 LRVTKAREG 0.001 acids, the end position for each peptide is the stmt position plus eight !TableXVII v.6-B35-9mers: 213P1F11 193 Table XVIII v. l-B35-10mers: 213P1F11 iTable XVIII v. l-B35-10mers: 213P1F11 Pos 1234567890 Score Pos 1234567890 Score 3 NPRSLEEEKY 180.000 Portion of 181 DQKGSCFIQT 0.300 Portion of RSLEEEKYDM 60.000 SEQ ID 93 GFLKGEDGEM 0.300 SEQ ID 0 NO: KAREGSEEDL 36.00 3; 27 160 PTYTDALHVY 0.200 NO: 3; each stall each stall 77 DPVSCAFWL 20.000 position is 55 DPTAEQFQEE 0.200 position is 123 KPKVYI IQAC 12.000 specified, 10 EKYDMSGARL 0.200 specified, 169 YSTVEGYIAY 10.000 the length 34 EDLDALEHMF 0.200 the length 37 DALEHMFRQL 6.000 of each 1 17 CQALRAKPKV 0.200 of each 118 QALRAKPKVY 6.000 peptide is 155 S PQTI PTYTD 0.200 peptide is amino 10 amino 159 I PTYTDALHV 6.000 56 PTAEQFQEEL 0.200 acids, the acids, the 232 QSTLRKRLYL 5.000 end 205 LLTEVTRRMA 0.200 end 14 MSGARLALI L 5.000 position for 78 PVSCAFWLM 0.200 position for EGSEEDLDAL 3.000 each 218 LVQEGKARKT 0.200 each 209 VTRRMAEAEL 3.000 peptide is 201 HILELLTEVT 0.200 peptide is the start 141 ETVGGDEIVM 3.000 107 NLFEALNNKN 0.200 the start position position 73 DSREDPVSCA 3.000 plus nine 185 SCFIQTLVDV 0.200 plus nine 204 ELLTEVTRRM 2.000 166 LHVYSTVEGY 0.200 231 IQSTLRKRLY 2.000 103 VKLENLFEAL 0.200 226 KTNPEIQSTL 2.000 178 YRHDQKGSCF 0.200 96 KGEDGEMVKL 1.800 43 FRQLRFESTM 0.200 • 16 GARLALI LCV 1.800 121 RAKPKVYI IQ 0.180 59 EQFQEELEKF 1.500 223 . ARKTN PEIQ 0.180 119 ALRAKPKVYI 1.200 . 149 VMVI KDS PQT 0.150 70 QAI DSREDPV 1.200 182 QKGSCFIQTL 0.100 157 QTI PTYTDAL 1.000 187 FIQTLVDVFT 0.100 85 VLMAHGREGF 1.000 186 CFIQTLVDVF 0.100 86 LMAHGREGFL 1.000 12 YDMSGARLAL 0.100 111 ALNNKNCQAL 1.000 194 VFTKRKGHIL 0.100 230 EIQSTLRKRL 1.000 39 LEHMFRQLRF 0.100 94 FLKGEDGEMV 0.900 100 GEMVKLENLF 0.100 143 VGGDEIVMVI 0.800 41 HMFRQLRFES 0.100 197 KRKGHILELL 0.600 15 SGARLALI LC 0.100 52 MKRDPTAEQF 0.600 23 LCVTKAREGS 0.100 ' 154 DS PQTI PTYT 0.500 210 TRRMAEAELV 0.090 79 VSCAFWLMA 0.500 215 , EAELVQEGKA 0.090 49 ESTMKRDPTA 0.500 133 RGEQRDPGET 0.090 176 IAYRHDQKGS 0.450 63 EELEKFQQAI 0.080 193 DVFTKRKGHI 0.400 228 N PEIQSTLRK 0.060 167 HVYSTVEGYI 0.400 212 RMAEAELVQE 0.060 183 KGSCFIQTLV 0.400 89 HGREGFLKGE 0.060 13 DMSGARLALI 0.400 135 EQRDPGETVG 0.060 153 KDS PQTI PTY 0.400 151 VI KDS PQTI P 0.060 138 DPGETVGGDE 0.400 33 EEDLDALEHM 0.060 150 MVI KDS PQTI 0.400 74 S REDPVSCAF 0.060 142 TVGGDEIVMV 0.300 104 ■ KLENLFEALN 0.060 110 EALNNKNCQA 0.300 6 SLEEEKYDMS 0.060 196 TKRKGHILEL 0.300 1 MSNPRSLEEE 0.050 113 NNKNCQALRA 0.300 184 GSCFIQTLVD 0.050 19 LALI LCVTKA 0.300 87 MAHGREGFLK 0.045 102 MVKLENLFEA 0.300 25 VTKAREGSEE 0.045 99 DGEMVKLENL 0.300 130 QACRGEQRDP ' 0.045 Table XVIII v.2-B35-10mers: 213P1F11 Table XVIII v.2-B35-10mers: 213P1F11 Pos 1234567890 Score 51 WWMCSRRGKD 0.001 33 QDTSPTDMIR 0.001 28 PLWNSQDTSP 0.001 16 PLYLPSEAPP 0.001 LHVYSTVEGP 0.001 50 PWWMCSRRGK 0.000 195 Table XVIII v.3-B35-10mers: 213P1F11 Pos 1234567890 Score 12 LPSPFPYLSL 20.000 Portion of 9 GATLPSPFPY 6.000 SEQID NO: 7; ATLPSPFPYL 1.000 each start 3 IIQACRGATL 1.000 position is QACRGATLPS 0.300 specified, 7 CRGATLPSPF 0.100 the length 11 TLPSPFPYLS 0.100 of each RGAT 0.100 peptide is 2 YIIQAC amino 6 ACRGATLPSP 0.030 acids, the 8 RGATLPSPFP 0.020 end 4 IQACRGATLP 0.010 position for 1 VYIIQACRGA 0.010 each peptide is the start position plus nine Table XVIII v.4-B35-10mers: 213P1F11 Table XVIII v.4-B35-10mers: 213P 1F11 Portion of SEQ ID NO: 9; each start position is specified, the length of each peptide is 10 amino acids, the end position for| each peptide is the start position plus nine 197 Table XVIII v.5-B35-10mers: 213P1F11 Table XVIII-v.6-B35-10mers: 213P1F11 Pos 1234567890 Score 9 AMNNKNCQAL 1.000 Portion of 1 VKLENLFEAM 0.400 SEQ ID 8 EAMNNKNCQA 0.300 NO: 13; each start NLFEAMNNKN 0.200 position is 2 KLENLFEAMN 0.060 specified, 3 LENLFEAMNN 0.015 the length 4 ENLFEAMNNK 0.010 of each MNNKNCQALR 0.010 peptide is amino 6 LFEAMNNKNC 0.003 acids, the 7 FEAMNNKNCQ 0.001 end position for each peptide is the start position plus nine TABLE XIXA: MHC Class I Analysis of Q13P1F11 v. 1: HLA-A*0201 nonamers 213P1F11 (9-mers). Listed are scores which Pos 1 2 3 4 5 6 7 8 9 score correlate witli tlve ligation strengtli to a defined HLA 22 . I L C V T K A R E 14 type for a sequence of amino acids. The algoritluns 50 s T M K R D P T A 14 used are based on (lie book "MHC Ligands and 85 • V L M A H G R E G 14 Peptide Motifs" by H.G.Rammensee, J.Bachmann 95 L G E D G E M V 14 and S.Stevanovic. The probability of being 1 12 L N N K N C Q A L 14 processed and presented is given in order to predict 187 F I Q T L V D V F 14 T-cell epitopes. 198 R K G H I L E L L 14 210 T R R M A E A E L 14 Table XIXA, part 1: MHC Class I nonamer 227 T N P E I Q S T L 14 analysis of 213P1F11 v. l (aa 1-242) 1 1 K Y D M s G A R L 13 213P1F11 v. 1 : HLA-A*0201 nonamers 19 L A L I L C V T K 13 Pos 1 2 3 4 5 6 7 8 9 score 28 A R E G S E E D L 13 201 H I L E L L T E V 29 Portion of 41 H M F R Q L R F E 13 A L I L C V T K A 24 SEQ ID 78 P V S C A F V V L 13 104 K L E N L F E A L 22 ;NO: 3; each| stail 127 Y I I Q A C R G E 13 158 T I P T Y T D A L 22 position is 160 P T Y T D A L H V 13 13 D M S G A R L A L 21 specified, 163 T D A L H V Y S T 13 38 A L E H M F R Q L 21 the length 165 A L H V Y S T V E 13 17 A R L A L I L C V 20 o each 190 T L V D V F T K R 13 18 R L A L I L C V T 20 peptide is 9 204 E L L T E V T R R 13 71 A I D S R E D P V 20 amino 212 R A E A E L V Q 13 233 S T L R K R L Y L 20 acids, the 214 A Ξ A E L V 2nd position Q. E G 13 1 18 Q A L R A K P K V 19 for each 79 V S C A F V V L M 12 121 R A K P K V Y I I 19 peptide is 80 S C A F V V L M A 12 151 V I K D S P Q T I 19 the start 1 19 A L R A K P K V Y 12 197 K R K G H I L E L 19 position 128 I I Q A C R G E Q 12 205 L L T E V T R R M 19 plus eight 140 G E T V G G D E I 12 6 S L E E E K Y D 18 157 Q T I P T Y T D A 12 94 F L K G E D G E M 18 171 T V E G Y I A Y R 12 107 N L F E A L N N K 18 189 Q T L V D V F T K 12 1 1 1 A L N N K N C Q A 18 218 L V Q E G K A R 12 143 V G G D E I V M V 18 .231 I Q S T L R K R L 12 183 K G S C F I Q T L 18 27 K A R E G S E E D 1 1 186 C F I Q T L V D V 18 35 D L D A L E H M F 1 1 202 I L E L L T E V T 18 74 S R E D P V S C A 1 1 226 K T N P E I Q S T . 18 135 E Q R D P G E T V 1 1 31 G S E E D L D A L 17 150 M V I K D S P Q T 1 1 ■ 97 G E D G E M V K L 17 170 S T V E G Y I A Y 11 100 G E M V K L E N L 17 175 Y I A Y R H D Q K 11 164 D A L H V Y S T V 17 211 R R M A E A E L V 11 S G A R L A L I L 16 230 E I Q S T L R K R 11 57 T A E Q F Q E E L 16 81 C A F V V L M A H 10 64 E L E K F Q Q A I 16 142 T V G G D E I V M 10 87 M A H G R E G F L 16 149 V M V I K D s P Q 10 195 F T K R K G H I L 16 167 ■ H V Y S T V E G Y 10 223 K A R K T N P E I 16 180 H D Q K G S C F I 10 86 L M A H G R E G F 15 184 G S C F I Q T L V 10 103 V K L E N L F E A 15 200 G H I L E L L T E 10 120 L R A K P K V Y I 15 216 A E L V Q E G K A 10 141 E T V G G D E I V 15 12 Y D M S G A R L A 9 144 G G D E I V M V I 15 16 G A R L A L I L C 9 234 T L R K R L Y L Q 15 44 R Q L R F E s T M 9 14 M S G A R L A L I 14 45 ,Q L R F E S T M K 9 21 L I L C V T K A R 14 46 L R F E S T M K R 9 199 E13P1F11 v. 1: HLA-A*0201 nonamers 213P1F11 v. 1: HLA-A*02 1 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 76 E D P V S C A F V 9 23 L C V T K A R E G 5 77 D P V S c A F V V 9 32 S Ξ E D L D A L Ξ 5 123 K P K V Y . I I Q A 9 73 D S R E D P V S C 5 148 I V M V I K D s P 9 82 A F V V L M A H G 5 153 K D S P Q T I P T 9 108 L F E A L N N N 5 162 Y T D A L H V Y S 9 1 16 N C Q A L R A K P . 5 168 V Y S T V E G Y I 9 161 T Y T D A L H V Y 5 176 I A Y R H D Q K G 9' 174 G Y I A Y R H D Q •5 206 L T E V T R R M A 9 178 Y R H D Q K G S C 5 213 M A E A E L V Q Ξ 9 193 D V F T K R K G H 5 219 V Q E G K A R K T 9 7 L E E E K Y D M S 4 2 S N P R S L E E E 8 24 C V T K A R E G S 4 34 E D L D A L E H M 8 26 T K A R E G S E E 4 51 T M K R D P T A E 8 30 E G S E •.E D L D A 4 52 M K R D P T A E Q 8 36 L D A L E H M F R 4 60 Q F Q . E E L E K F 8 69 Q Q A I D S R E D 4 125 K V Y I I Q A C R 8 102 M V K L E N L F E 4 147 E, I V M V I K D S 8 110 E A L N N K N C Q 4 191 L V D V F T K R K 8 129 I Q A C R G E Q R 4 194 V F T K R K G H I 8 131 A C R G E Q R D P 4 203 L E L L T E V T R 8 136 Q R D P G E T V G 4 208 E V T R R M A E A 8 182 Q K G S C F I Q T 4 37 D A L E H M F R Q 7 3 • N P R S L E E E K 3 43 F R Q L R F E S T 7 5 R S L E E E K Y D 3 56 P T A E Q F Q E E 7 39 L E H M F R Q L R 3 67 K F Q Q A I D S R 7 42 M F R Q L R F E S 3 70 Q A I D S R E D P 7 53 K R D P T A E Q F ' 3 83 F V V L M A H G R 7 72 I D S R E D P V S 3 84 V V L M A H G R E 7 75 R E D P V S C A F 3 89 H G R E G F L K G 7 96 K G E D G E M V K 3 90 G R E G F L K G E 7 126 V Y I I Q A C R G 3 101 E M V K L E N L F 7 152 I K D S P Q T I P 3 134 G E Q R D P G E T 7 61 F Q E E L E K F Q 2 137 R D P G E T V G G 7 65 L E K F Q Q A I D 2 146 D E I V M V I K D 7 88 A H G R E G F L K 2 166 L H V Y S T V E G 7 91 R E G F L K G E D 2 188 I Q T L V D V F T 7 98 E D G E M V K L E 2 199 K G H I L E L L T 7 109 F E A L N N K N C 2 217 E L V Q E G K A R 7 122 A K P K V Y I I Q 2. 222 G K A R K T N P E 7 132 C R G E Q R D P G 2 1 M S N P R S L E E 6 145 G D E I V M V I K 2 V T K A R E G S E 6 156 P Q T I P T Y T D 2 63 E E L E K F Q Q A 6 159 I P T Y T D A L H 2 93 G F L K G E D G E 6 177 A Y R H D Q K G S 2 105 L E N L F E A L N 6 192 • V 0 V F T K R K G 2 1 14 N K N C Q A L R A 6 196 T K R K G H I L E 2 115 K N C Q A L R A K 6 229 P E I Q S T L R K .2 130 Q A C R G E Q R D 6 48 F E S T M K R D P 1 138 D P G E T V G G D 6 49 E S T M K R D P T 1 154 D S P Q T I P T Y 6 58 A E Q F Q E E L E 1 155 S P Q T I P T Y T 6 59 E Q F Q E- E L E K 1 169 Y S T V E G Y I A 6 68 F Q Q A I D S R E 1 185 S . C F I Q T L V D 6 99 ' D G E M V K L E N 1 209 V T R R M A E A E 6 1 13 N N K N C Q A L R 1 200 213P1F11 v.1: HLA-A*0201 nonamers B13P1F11 v.l: HLA-A1 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 117 C Q A L R A K P K 1 139 P G Ξ T V G G D E 13 124 P K V Y. I I Q A C 1 28 A R Ξ G S E E D L 12 133 R G E Q R D P G E 1 35 D L D A L E H M F 12 172 ' V E G Y I A Y R H 1 80 S C A F V V L M A 12 179 R H. D Q K G S C F 1 144 G G D E I V M V I 12 207 T E V T R R M A E 1 145 G D E I V M V I K 12 215 E A E L V Q E G K 1 .160 • P T Y T D A L H V 12 220 Q E G K A R K T N 1 171 T V E G Y I A Y R 12 224 A R K T N P E I Q 1 202 I L E L L T E V T 12 4 P R S L E E E K Y -1 215 E A E L V Q E G K 12 8 E E E K Y D M S G -1 7 L E E E K Y D M s 11 62 Q E E L E K F Q Q -1 8 E E E K Y D M S G 11 92 E G F L K G E D G -1 11 K Y D M S G A R L 11 106 E N L F E A L N N -1 56 P T A E Q F Q E E 11 228 N P E I Q S T L R -1 61 F Q E E L E K F Q 11 232 Q S T L R K R L Y -1 64 E L E K F Q Q A I 11 40 E H M F R Q L R F -2 71 A I D S R E D P V 11 47 R F E S T M K R D -2 90 G R E G F L K G E 11 55 D P T A E Q F Q E -2 152 I K D S P Q T I P 11 66 E K F Q Q A I D S -2 179 R H D Q K G S C F 11 173 E G Y I A Y R H D -2 185 S C F I Q T L V D 11 139 P G E T V G G D E -4 47 R F E S T M K R D 10 221 E G K A R K N P -4 57 T A E Q F Q E E L 10 62 Q E E L E K F Q Q 10 213P1F11 v.l: HLA-A1 nonamers 133 R G E Q R D P G E 10 Pos 1 2 3 4 5 6 7 8 9 score 153 K D S P Q T I P T 10 170 S T V E G Y I A Y 29 Portion of 157 Q T I P T Y T D A 10 232 Q S T L R K R L Y 21 SEQID 191 L V D V F T K R K 10 154 D S P Q T I P T Y 19 NO: 3; each 213 M A E A E L V Q E 10 start 99 D G E M V K L E N 18 226 K T N P E I Q S T 10 position is 119 A L R A K P K V Y 18 141 E T V G G D E I V 9 specified, 162 Y T D A L H V Y S 18 A L the length 13 D M S G A R L 8 206 • L .E V T R R M A 18 of each 15 S G A R L A L I L 8 4 P R S L E E E K Y 17 peptide is 9 50 S T M R D P T A 8 136 Q R D P G E T V G 17 amino 79 V S C A F V V L M 8 33 E E D L D A L E H 16 acids, the 114 N K N C Q A L R A 8 75 R E D P V S C A F 16 end position 190 T L V D V F T K R 8 for each 161 T 'Y D A L H V Y 16 195 F T K R K G H I L 8 peptide is 167 H V Y S T V E G Y 16 T K R K G H .1 L E the start 196 . 8 38 A L E H M F R Q L 15 position 199 K G H I L E L L T 8 233 S T L R K R L Y L 15 plus eight 17 A R L A L I L C V 7 1 M S N P R S L E E 14 25 V T K A R E G S E 7 31 G S E E D L D A L 14 102 M V K L E N L F E 7 32 S E E D L D A L E .14 122 A K P K V Y I I Q 7 53 K R D P T A E Q F 14 146 D E I V M V I K D 7 74 S R E D P V S C A 14 169 Y S T V E G Y I A 7 104 K L E N L F E A L 14 182 Q K G S C F I Q T 7 219 V Q E G K A R K T 14 189 Q T L V D V F T K 7 228 N P E I Q S T L R 14 197 K R K G H I L E L 7 6 S L E E E K Y D M 13 209 V T R R M A E A E 7 89 H G R E G F .L K G 13 212 R M A E A E L V Q 7 96 K G E D G E M V K 13 12 Y D S G A R L A 6 97 G E D G E M V K L 13 14 M S G A R L A L I 6 108 L F E A L N N K N 13, 16 G A R L A L I L c 6 201 Q13P1F11 v.l: HLA-A1 nonamers E13P1F11 v.l: HLA-A1 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score E G S E E D L D A 6 109 F E A L N N K N C 2 40 E H M F R Q L R F 6 113 N N N C Q A L R 2 59 E Q F Q E E L E K 6 118 Q A L R A K P K V 2 66 E K F Q Q A I D s 6 127 Y I I Q A C R G E 2 106 E N L F E A L N N 6 130 Q A c R G E Q R D 2 142 T V G G D E I V M 6 131 A C R G E Q R D P 2 184 G S C F I Q T L V 6 135 E Q R D P G E T V 2 200 G H I L E L L T E 6 143 V G G D E I V M V 2 229 P E I Q S T L R K 6 147 E I V M V I K D S 2 A L I L C V T K A 5 155 S P Q T I P T Y T 2 49 E S T M K R D P T 5 159 I P T Y T D A L H 2 58 A E Q F Q E E L E 5 164 D A L H V Y S T V 2 121 R A K P K V Y I I 5 ' 174 G Y I A Y R H D Q 2 123 K P K V Y I I Q A 5 175 Y I A Y R H D Q K 2 216 A E L V Q E G K A 5 176 I A Y R H D Q K G 2 225 R K T N P E I Q S 5 180 H D Q K G S c F I 2 R S L E E E K Y D 4 188 I Q T L V D V F T 2 43 F R Q L R F E S T 4 198 R K G H I L E L L 2 73 D S R E D P V S C 4 204 E L L T E V T R R 2 78 P V S C A F V V L 4 211 R R M A E A E L V 2 88 A H G R E G F L K 4 214 A E A .E L V Q E G 2 165 A L H V Y S T V E 4 217 E L V Q E G K A R 2 178 Y R H D Q K G S C 4 218 L V Q E G K A R K 2 39 L E H M F R Q L R 3 230 E I Q S T L R K R 2 63 E E L E K F Q Q A 3 22 I L c V T K A R E 1 85 V L M A H G R E G 3 24 C V T K A R E G S 1 86 L M A H G R E G F 3 26 T K A R E G S E E 1 94 F L K G E D G E M 3 42 M F R Q L R F E S 1 98 E D G E M V K L E 3 48 F E S T. M K R D P 1 1 11 A L N N K N C Q A 3 52 M K R D P T A E Q 1 1 16 N C Q A L R A K P 3 68 F Q Q A I D S R E 1 126 V Y I I Q A C R G 3 70 Q A I D s R E D P 1 132 C R G E Q R D P G 3 72 I D s R E D P V S 1 140 G E T V G G D E I 3 77 D P V S C A F V V ] 168 V Y S T V E G Y I 3 82 A F V V L M A H G 1 187 F I Q T L V D V F 3 83 F V V L M A H G R 1 192 V D V F T K R K G 3 84 V V L M A H G R E 1 205 L L T E V T R R M 3 87 M A H G R E G F L 1 234 T L R K R L Y L Q 3 92 E G F L K G E D G 1 2 S N P R S L E E E 2 103 V K L E N L F E A 1 18 R L A L I L C V T 2 107 ■ N L F E A L N N 1 21 L I L C V T K A R 2 120 L R A K P K V Y I 1 27 K A R E G S E E D 2 128 I I Q A C R G E Q r 29 R E G S E E D L D 2 129 I Q A C R G E Q R 1 41 H M F R Q :L R F E 2 137 R D P G Έ T V G G 1 45 Q L R F E s T M K 2 138 D P G E- T V G G D 1 46 L R F E S T M K R 2 149 V M V I K D S P Q 1 60 Q F Q E E L E K F 2 151 V I K D S P Q T I 1 65 L E K F Q Q A I D 2 156 P Q T I P T Y T D 1 81 C A F V V •L M A H 2 158 T I P T Y T D A L 1 93 G F L K G E D G Ξ 2 163 T D A L H V Y S T 1 95 L K G E D G E M V 2 172 V E G Y I A Y R H 1 101 E M V K L E N L F 2 177 A Y R H D Q K G S 1 105 L E N L F E A L N 2 181 D Q K G S C F I Q 1 202 213P1F11 v.l: HLA-A1 nonamers P13P1F11 v.l: HLA-A26 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 193 D'V F T K R K G H 1 100 G E M V K L E N L 14 194 V F T K R K G H I 1 127. Y I I Q. A C R G E 14 207 T E V T R R M A E 1 151 V I K D S P Q T I 14 220 Q E G K A R K T N 1 179 R H D Q K G S C F 14 223 K A R K T N P E I 1 186 C F I Q T L V D V 14 224 A R K T N P E I Q 1 189 Q T L V D V F T K 14 231 I Q S T L R K R L 1 190 T L V D V F T K R 14 218 L V Q E G K A R K 14 213P1F11 v.l: HLA-A26 nonamers | 9 E E K Y D M S G A 13 Pos 1 2 3 4 5 6 7 8 9 score 18 R L A L I L C V T 13 D L D A L E H M F 27 Portion of 37 D A L E H M F R Q 13 170 S T V E G Y I A Y 27 SEQID 79 V S C A F V V L M 13 167 H V Y S T V E G Y 26 NO: 3; each 82 A F V V L M A H G 13 start 187 F I Q T L V D V F 25 138 D P G E T V G G D 13 position is 60 Q F Q E E L E K F 23 ' 146 D E I V M V I K D 13 specified, 78 P V S C A F V V L 23 the length 162 Y T D A L H V Y S 13 154 D S P Q T I P T Y 23 of each 183 K G S C F I Q T L 13 104 K L E N L F E A L 22 peptide is 9 234 T L R K- R L Y L Q 13 208 E V T R R M A E A 22 amino 11 K Y .D M S G A R L 12 230 E I Q S T L R K R ' 22 acids, the 24 C V T K A R E G S 12 E H M F R Q L end position 38 A L 21 25 V T K A R E G S E 12 for each 34 E D L D A L E H M 20 47 R F E S T M K R D 12 peptide is 56 P T A E Q F Q E E 20 50 S T M K R D P T A 12 the start 94 F L K G E D G E M 20 position 67 K F Q Q A I D S R 12 142 T V G G D E I V M 20 plus eight 71 A I D S R E D P V 12 147 E I V M V I K D S 20 . 86 L M A H G R E G F 12 158 T I P T Y T D A L 20 98 E D G E M V K L E 12 193 D V F T K R K G H 20 175 Y I A Y R H D Q K 12 195 F T K R K G H I L 20 191 L V D V F T K R K 12 119 A L R A K P K V Y 19 198 R K G H I L E L L 12 204 E L L T E V T R R 19 227 T N P E I Q S T L 12 205 L L T E V T R R M 19 4 P R S L E E E K Y 11 233 S T L R K R L Y L 19 21 L I L C V T K A R 1 6 S L E E E K Y D M 18 28 A R E G s E E D L 1 101 E M V K L E N L F 18 83 F V V L M A H G R 11 141 E T. V G G D E I V 18 84 V V L M A H G R E 11 157 Q T I P T Y T D A 18 102 M V K L E N L F E 1] 201 H I L E L L T E V 18 112 L N N K N C Q A. L 11 226 K T N P E I Q S T 18 125 K V Y I I Q A C R 11 13 D M S G A R L A L 17 128 I I Q A C R G E Q 11 40 E H M F R 'Q L R F 17 148 I V M V I K D S P 11 64 E L E K F Q Q A I 17 160 P T Y T D A L H V 11 97 G E D G E M V K L 17 164 D A L H V Y S T V 11 107 N L .F E A L N N K 17 173 . E G Y I A Y R H D 11 171 T V E G Y I A Y R 17 206 L T E V T R R M A 11 197 K R K G H I L E L 16 209 V T R R M A E A E 11 217 E L V Q E G K A R 16 15 S G A R L A L I L 10 A L I L C V T K A 15 22 I L C V T K A R E 10 31 G S E E D L D A L 15 57 T A E Q F Q E E L 10 63 E E L E K F Q Q A 15 5 E Q F Q E E L E K 10 150 M V I K D S P Q T 15 66 E K F Q Q A I D S 10 161 T Y T D A L H V Y 15 73 D S R E D P V S C 10 53 K R D P T A E Q F 14 87 M A H G R E G F L 10 75 R E D P V S C A F 14 111 A L N N K N C Q A 10 203 E13P1F11 v.l: HLA-A26 nonamers E13P1F11 v.l: HLA-A26 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 181 D Q K G S C F I. Q 10 145 G D E I V M V I K 5 210 T R R M A E A E L 10 229 P E I Q S T L R K 5 214 A E A E L V Q E G 10 16 G A R L A L I L c 4 231 I Q S T L R K R L 10 70 Q A I D S R E D P 4 232 Q s T L R K R L Y 10 126 V Y I I Q A C R G 4 8 E E E K Y D M S G 9 219 V Q E G K A R K T 4 42 M F R Q L R F E S 9 1 M S N P R S L E E 3 44 R Q L R F E S T M 9 3 N P R S L E E E K 3 45 Q L R F E S T M K 9 26 T K A R E G S E E 3 76 E D P V S C A F V 9 27 K A R E G S E E D 3 85 V L M A H G R E G 9 32 s E E D L D A L E 3 92 E G F L K G E D G 9 52 M K R D P T A E Q 3 202 I L E L L T E V T 9 61 F Q E E L E K F Q 3 7 L E E E K Y D M S 8 65 L E K F Q Q A I D 3 E K Y D M S G A R 8 95 L K G E D G E M V 3 E G S E E D L D A 8 116 N C Q A L R A K P 3 33 E E D L D A L E H 8 120 L R A K P K V Y I 3 81 C A F V V L M A H 8 132 C R G E Q R D P G 3 93 G F L K G E D G E 8 133 R G E Q R D P G E 3. 108 L F E A L N N K N 8 136 Q R D P G E T V G 3 121 R A K P K V Y I I 8 153 K D S P Q T I P T 3 144 G G D E I V M V I 8 166 L H V Y S T V E G 3 165 A L H V Y S T V E 8 178 Y R H D Q K G S C 3 194 V F T K R K G H I 8 185 S C F I Q T L V D 3 221 E G K A R K T N P 8 212 R M A E A E L V Q 3 2 ' S N P R S L E E E 7 223 K A R K T N P E I 3 41 H M F R Q L R F E 7 5 R S L E E E K Y D 2 46 L R F E S T M K R 7 19 L A L I L C V T K 2 49 E S T M K R D P T 7 23 L C V T K A R E G 2 55 D P T A E Q F Q E 7 36 L D A L E H M F R 2 77 D P V S C A F V V - 7 51 T M K R D P T A E 2 89 H G R E G F L K G 7 54 R D P T A E Q F Q 2 90 G R E G F L K G E 7 68 F Q Q A I D S R E 2 99 D G E M V K L E N 7 69 Q Q A I D S R E D 2 103 V K L E N L F E A 7 91 R E G F L K G E D 2 106 E N L F E A L N N 7 96 K G E D G E M V K 2 135 E Q R D P G E T V 7 109 F E A L N N K N C 2 143 ■ V G G D E I V M V 7 113 N N K -N C Q A L R 2 215 E A E L V Q E G K 7 114 N K N •C Q A L R A 2 74 S R E D P V S C A 6 118 Q A L R A K P K V 2 80 S C A F V V L M A 6 129 I Q A C R G E Q R 2 110 E A L N N K N C Q 6 130 Q A C R G E Q R D 2 1 15 K N C Q A L R A K 6 131 A C R G E Q R D P 2 122 A K P K V Y I I Q 6 149 V M V I K D S P Q 2 123 .. K P K V Y I I Q' A 6 152 I K D S P Q T I P 2 124 P K V Y I I Q A, C 6 174 G Y I A Y R H D Q 2 163 T D A L H V Y S T 6 176 I A Y R H D Q K G . 2 182 Q K G S C F I Q T 6 207 T E V T R R M A E 2 200 G H Ί L E L L T E 6 216 A E L V Q E G K A 2 213 M A E A E L V Q E 6 222 G K A R K T N P E 2 14 M S G A R L A L I 5 12 Y D M S G A R L A 1 17 A R L A L I L C V 5 29 R E G S E E D L D 1 43 F R Q L R F E S T 5 39 L E H M F R Q L R 1 137 R D P G E T V G G 5 58 A E Q F Q E E L E 1 204 1F11 v.l : HLA-A26 nonamers P13P1F11 v.l: HLA-A3 nonamers Pos 1 2 3 4 5 6 7 8 9 score 150 M V I K D S P Q T 16 187 F I Q T L V D V F 16 193 D V F T K R K G H 16 229 P E I Q S T L R K 16 22 I- L C V T K A R E 15 85 V L M A H G R E G 15 102 M V K L E L F E 15 104 K L E N L F E A L 15 129 I Q A C R G E Q R 15 151 V I K D S P Q T I 15 179 R H D Q K G S c F 15 203 L E L L T E V T R 15 234 T L R K R L Y L Q 15 94 F L K G E D G E M 14 1 15 K N C Q A L R A K 14 128 I I Q A C R G E Q 14 148 I V M V I K D S P 14 3 N P R S L E E E K 13 59 E Q F Q E E L E K 13 71 A I D S R , E D P V 13 75 R E D P V S C A F 13 84 V V L M A H G R E 13 212 R M A E A E L V Q 13 6 S L E E E K Y D M 12 136 Q R D P G E T V G 12 201 H I L E L L T E V 12 205 L L T E V T R R M 12 215 E A E L V Q E G K 12 230 E I Q S T L R K R 12 E K Y D M S G A R 11 S G A R L A L I L 1 1 24 C V T K A R E G S 1 1 33 E E D L D A L E H 1 1 64 E L E K F Q Q A I 1 1 73 D S R E D P V S c 1 1 127 Y I I Q A C R G E 1 1 135 E Q R D P G E T V 11 161 T Y T D A L H V Y 1 1 170 S T V E G Y I A Y 1 1 200 G H I L E L L T E 11 232 Q S T L R K R L Y 1 1 233 S T L R K R L Y L 1 1 17 A R L A L I L c V 10 26 T K A R E G S E E 10 137 R D P G E T V G G 10 154 D S P Q T I P T Y 10 160 P T Y T D A L H V 10 164 D A L H V Y S T V 10 209 V T R R M A E A Ξ 10 11 K Y D M S G A R L 9 27 K A R E G S E E D 9 40 E H M F R Q L R F ' 9 67 K F Q Q A I D S R 9 72 I D s R E D P V S 9 205 E13P1F11 v.l: HLA-A3 nonamers E13P1F11 v.l: HLA-A3 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 89 H G R E G F L K G 9 144 G G D E I V M V I 6 113 N N K N C Q A L R 9 153 K D S P Q T I. P T 6 121 R A K P K V Y I I 9 162 Y T D A L H V Y s 6 147 E I V M V I K D s 9 174 G Y I A Y R H D Q 6 157 Q T I P T Y T D A 9 177 A Y R H D Q K G s 6 158 T I P T Y T D A L 9 210 T R R M A E A E L 6 159 I P T Y T D A L •H 9 216 A E L V Q E G K A 6 176 I A Y R H D Q K G 9 219 V Q E G K A R K T 6 185 s C F I Q T L V D 9 221 E G K A R K T N P 6 197 K R K G H I L E L 9 223 K A R K T N P E I 6 211 R R M A E A E L V 9 228 N P E I Q S T L R 6 213 M A E A E' L V Q E 9 12 Y D M S G A R L A 5 1 M S N P R S L E E 8 28 A R E G S E E D L 5 V T K A R E G S E 8 32 ■ s E E D L D A L E 5 86 L M A H G R E G F 8 39 L E H M F R Q L R 5 106 . E N L F E A L N N 8 43 F R Q L R F E S T 5 131 A C R G E Q R D P 8 55 D P T A E Q F Q E 5 188 I Q T L V D V F T 8 68 F Q Q A I D S R E 5 195 F T K R K G H I L 8 74 S R E D P V S C A 5 225 R K T N P E I Q S 8 87 M A H G R E G F L 5 226 K T N P E I Q S T 8 91 R E G F L K G E D 5 227 T N P E I Q s T L 8 92 E G F L K G E D G 5 4 P R S L E E E K Y 7 95 L K G E D G E M V 5 R S L E E E K Y D 7 120 L R A K P K V Y I 5 13 D M S G A R L A L 7 134 G E Q R D P G E T 5 14 M S G A R L A L I 7 139 P G E T V G G D E 5 36 , L D A L E H M F R 7 140 G Ξ T V G G D E I 5 50 S T M K R D P T A 7 149 V M V I K D S P Q 5 51 T M K R D P T A E 7 156 P Q T I P T Y T D 5 52 M K R D P T A E Q 7 173 E G Y I A Y R H D 5 60 Q F Q E E L E K F 7 182 ' Q K G S C F I Q T 5 63 E Ξ L E K F Q Q A 7 196 T K R K G H I L E 5 70 Q A I D s R E D P 7 207 T E V T R R M A E 5 77 D P V S c A F V V 7 214 A Ξ A E L V Q Έ G 5 80 S C A F V V L M A 7 222 G K A R K T N P Ξ 5 81 C A F V V L M A H 7 224 A R K T N P E I Q 5 118 Q A L R A K P K V 7 2 S N P R S L E E E 4 123 - K P K V Y I I Q A 7 9 E E K Y D M S G A 4 126 V Y I I Q A C R G 7 16 G A R L A L I L C 4 130 Q A c R G E Q R D 7 23 L C V T K A R E G 4 172 V E G Y I A Y R H 7 29 R Ξ G S E E D L D 4 183 K G S C F I Q T L . 7 ■ 30 E G S E E D L D A 4 186 c F I Q T L V D V 7 42 M F R Q L R F E S 4 199 K G H I L E L L T 7 47 R F E S T M K R D 4 220 Q E G K A R K T N 7 54 R D P T A E Q F Q " 4 46 L R F E S T M K R 6 65 L E K F Q Q A I D 4 62 Q E E L E K F Q Q 6 69 Q Q A I D S R E D 4 79 V S C A F V V L M 6 93 G F L K G E D G E 4 82 A F V V L M A H G 6 99 D G E M V K L E N 4 97 G Ξ D G E M V K L 6 101 E M V K L E N L F 4 114 N K N C Q A L R A 6 105 L Ξ N L F E A L N 4 116 N c Q A L R A K P 6 122 A K P K V Y I I Q 4 133 R G E Q R D P G E 6 141 E T V G G' D E I V 4 143 V G G D E I V M V 6 163 T D A L H V Y s - T 4 206 E13P1F11 v.l: HLA-A3 nonamers bl3PlFll v.l : HLA-B*0702 nonan ers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 166 L H V Y S T V E G 4 13 . D M S G A R L A L 18 Portion of | 178 Y R H D Q K G S . C 4 77 D P V S C A F V V 17 SEQ ID 198 R K G H I L E L L 4 123 K P K V Y I I Q A 17 NO: 3; each| L E E E K Y D M S stiirt 7 3 155 S P Q T I P T Y T 17 position is 8 E Ξ E K Y D M S G 3 78 P V s C A F V V L 16 specified, 34 E D L D A L E H M 3 97 G E D G E M V K L 15 the length 37 D A L E H M F R Q 3 197 K R K G H I L E L 15 of each 41 H M F R Q L R F E 3 1 1 K Y D M S G A R L 14 peptide is 9 58 A E Q F Q E E L E 3 28 A R E G S E E D L 14 amino 66 . E K F Q •Q A I D S 3 231 I Q S T L R K R L 14 acids, the E D P V S C A F V 3 15 S G A R L A L I L 13 ]end position| 76 for each 103 V K L E N L F E A 3 38 A L E H M F R Q L 13 peptide is 1 10 E Ά L N N K N C Q 3 87. M A H G R E G F L 13 the start 146 D E I V M V I K D 3 104 K L E N L F E A L 13 position 152 I K D S P Q T I P 3 120 L R A K P K V Y I 13 plus eight 155 S P Q T I P T Y T 3 135 E Q R D P G E T V 13 169 Y s T V E G Y I A 3 183 K G S C F I Q T L 13 181 D Q K G S C F I Q 3 210 T R R M A E A E L 13 194 V F T K R K G H I 3 233 s T L R K R L Y L 13 231 I Q s T L R K R L 3 3 N P R S L E E E K 12 31 G s E E D L D A L 2 1 12 L N N K N C Q A L 12 49 E s T M K R D P T 2 138 D P G E T V G G D 12 56 P T A E Q F Q E E 2 153 K D S P Q T I P T 12 90 G R E G F L K G E 2 158 T I P T Y T D A L 12 108 L F E A L N N K N 2 159 I P T Y T D A L H 12 109 F E A L N N K ■N C 2 198 R K G H I L E L L 12 132 C R G E Q R D P G 2 17 A R L A L I L C V 1 1 138 D P G E T V G G D 2 30 E G S E E D .L D A 1 1 168 V Y S T V E G Y I 2 31 G S E E D L D A L 1 1 180 H D Q K G S C F I 2 55 D P T A E Q F Q E 1 1 206 L T E V T R R M A 2 100 G E M V K L E N L 1 1 . 61 F Q E E L E K F Q 1 195 F T K R K G H I L 1 1 98 E D G E M V K L E 1 223 K A R K T N P E I 1 1 100 G E M V K L E N L 1 228 N P E I Q S T L R 1 1 1 12 L N N K N C Q A L 1 20 A L I L C V T K A 10 124 P K V Y I I Q A C 1 40 E H M F R Q L R F 10 184 G s C F I Q T L V 1 57 T A E Q F Q E E L 10 71 A I D S R E D P V 10 74 S R E D P V S C A 10 75 •R E D P V S C A F 10 79 V S C A F V V L M 10 80 S C A F V V L M A 10 142 T V G G D E I V 10 188 I Q T L V D V F T 10 227 T N P E I Q S T L 10 14 M S G A R L A L I 9 18 R L A L I L C V T 9 49 E S T M K R D P T 9 50 S T M K R D P T A 9 53 K R D P T A E Q F 9 64 E L E K F Q Q A I 9 76 E D P V S C A F V 9 121 R A K P K V Y I I 9 141 E T V G G D E I V 9 207 I213P1F11 v.l: HLAJB*0702 nonamers E13P1F11 v.l: HLA-B*0702 nonamers Pos 1 2 3 4 5 6 7 Θ 9 score Pos 1 2 3 4 5 6 7 8 9 score 143 V G G D E I V M V 9 33 E E D L D A L E H 4 144 G G D E I V M V I 9 73 D S R- E D P V S C 4 179 R H D Q K G S C F 9 88 A H G R E G F L K 4 199 K G H I L E L L T 9 136 Q R D P G E T V G 4 202 I L E L L T E V T 9 156 P Q T I P T Y T D 4 211 R R M A E A E L V 9 162 Y T D A L H V Y S 4 D L D A L E H M F 8 165 A L H V Y S T V E 4 63 E E L E K F Q Q A 8 177 A Y R H D Q K G S 4 86 L M A H G R E G F 8 185 S c F I Q T L V D 4 94 F L K G E D G E M 8 214 A E A E L V Q E G 4 95 L K G E D G E M V 8 225 R K T N P E I Q S 4 101 E V K L E N L F 8 1 M. s N P R S L E E 3 11 1 A L N N K N C Q A 8 27 K. A R E G S E E D 3 1 14 N K N C Q A L R A 8 42 M F R Q L R F E S 3 150 M V I K D S P Q T 8 45 Q L R F E S T M K 3 157 Q T I P T Y T D A 8 59 E Q F Q E E L E K 3 160 P T Y T D A L H V . 8 82 A F V V L M A H G 3 163 T D A L H V Y S T 8 85 V L M A H G R E G 3 168 V Y S T V E G Y I 8 98 E D G E M V K L E 3 180 H D Q K G S C F I 8 102 M V K L E N L F Ξ 3 182 Q K G s C F I Q T 8 106 E N L F E A L N 3 186 C F I Q T L V D V 8 116 N C Q A L R A K P 3 187 F I Q T L V D V F 8 122 A K P K V Y I I Q 3 208 E V T R R M A E A 8 128 I I Q.A C R G E Q 3 216 A Ξ L V Q E G K A 8 129 I Q A C R G E Q R 3 219 V Q E G K A R K T 8 132 C R G E Q R D P G . 3 226 K 1, N P E I Q S T 8 166 L H V Y S T V E G 3 9 E E K Y D M S G A 7 171 T V E G Y I A Y R 3 12 Y D M S G A R L A 7 196 T K R K G H I L E 3 34 E D L D A L E H M 7 204 E L L T E V T R R 3 43 F R Q L R F E S T 7 209 V T R R M A E A E 3 44 R Q L R F E S M 7 213 M A E A E L V Q E 3 52 M K R D P T A E Q 7 217 E L V Q E G K A R 3 1 18 Q A L R A K P K V 7 220 Q E G K A R K T N 3 140 G E T V G G D E I 7 221 E G K A R K T N P 3 169 Y S T V. E G Y I A 7 222 G K A R K T N P E 3 184 G S C F I Q T L V 7 229 P E I Q S T L R K 3 194 V F T K R K G H I . 7 234 T L R K R L Y L Q 3 201 H I L E L L T Έ V 7 4 P R S L E E E K Y 2 205 L L T E V T R R M 7 8 E E E K Y D M S G 2 206 L T E V T R R M A 7 10 E K Y D M S G A R 2 6 S L E E E K Y D 6 16 G A R L A L I L C 2 60 Q F Q E E L E K F 6 19 L A L I L C V T K 2 103 V K L E N L F E A 6 21 L I L C V T K A R 2 119 A L R A K P K V Y 6 22 I L C V T K A R E 2 131 A C R G E Q R D P 6 24 C V T K A R E G S 2 134 G Ξ Q R D P G E T 6 26 T A R E G S E E 2 137 R D P G E T V G G 6 29 R E G S E E D L D 2 151 V I K D S P Q T I 6 36 . L D A L E H M F R 2 152 I K D S P Q T I P 6 48 F Ξ S T M K R D P 2 164 D A L H V Y S T V 6 51 T M K R D P T A E 2 72 I D S R E D P V s 5 54 R D P T A E Q F Q 2 89 H G R E G F L K G 5 56 P T A E Q F Q E E 2 212 R M A E A E L V Q 5 58 A E Q F Q E E L E 2 208 I213P1F11 v.l: HLA-B*0702 nonamers G13P1F11 v. l: HLA-B*0702 nonamcrs Pos 1 2 3 4 5 6 7 8 9 score 90 G R E G F L K G E 2 91 R E G F L K G E D 2 92 E G F' L K G E D G 2 96 K G E D G E M V 2 99 D G E M V K L E N ' 2 115 K N C Q A L R A K 2 117 C Q A L R A K P K 2 125 K V Y I I Q A C R 2 133 R G E Q R D P G E 2 145 ■ G D E I V M V I K 2 148 I V M V I K D S P 2 174 G Y I A Y R H D Q 2 181 D Q K G. S C F I Q 2 190 T L V D V F T K R 2 191 L V D V F T K R K 2 200 G H I L E L L T E 2 203 L E L L T E V T R 2 207 T Ξ V T R R M A E 2 224 A R K T N P E I Q 2 R S L E E E K Y D 1 V T K A R E G S E 1 32 S E E D L D A L E 1 39 L E H M F R Q L R 1 41 H M F R Q L R F E 1 46 L R F E S T M K R 1 47 R F E S T M K R D 1 61 F Q E E L E K F Q 1 65 L E K F Q Q A I D 1 66 E K F Q Q A I D S 1 67 K F Q Q A I D S R 1 68 F Q Q A I D S R E 1 69 Q Q A I D S R E D 1 70 Q A I D S R E D P 1 81 C A F V V L M A H 1 93 G F L K G E D G E 1 105 L E N L F E A L N 1 108 L F E A L N N K N 1 109 F E A L N N K N C 1 1 10 E A L N N K N C Q 1 113 N N K N C Q A L R 1 124 P K V Y I I Q A C 1 139 P G E T V G. G D E 1 146 D E I V M V I K D 1 1 7 E I V M V I ■K D S 1 149 V M V I K D S P Q 1 154 D S P Q T I P T Y 1 161 T Y T D A L H V Y 1 167 H V Y S T V E G Y 1 170 S T V E G Y I A Y 1 172 V E G Y I A Y R H 1 173 E G Y I A Y R H 0 1 175 Y I A Y R H D Q K 1 176 I A Y R H D Q K G 1 189 Q T L V D V F T K 1 209 I213P1F11 v.l: HLA-B*08 nonamei s Q13P1F11 v.l: HLA-B*08 nonamei s Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 78 P V S C A F V V L 11 215 E A E L V Q E G K 7 112 L N N K N C Q A L 11 21 L I L C V T K A R 6 149 ' V M V I K D s P Q 11 37 D A L E H M F R Q 6 183 K G S c F I Q T L 11 53 K R D P T A E Q F 6 193 D V F T K R K G H 1 1 55 D P T A E Q F Q Ξ 6 208 E V T R R M A E A 11 77 D P V S C A F V V 6 219 V Q E G K A R K T 11 81 C A F V V L M A H 6 222 G K A R K T N P E 1 1 86 L M A H G R E G F 6 6 S L E E E K Y D 10 98 E D G E M V K L E 6 1 1 K Y D M S G A R L 10 110 E A L N N K N C Q 6 28 A R E G S E E D L 10 128 I I Q A C R G E Q 6 43 F R Q L R F E S T 10 13 1 A C R G E Q R D P 6 50 S T M K R D P T A 10 190 T L V D V F T K R 6 65 L E K F Q Q A I D 10 196 T K R K G H I L E 6 113 N N K N C Q A L R 10 228 N P E I Q S T L R 6 117 C Q A' L R A K P 10 230 E I Q S T L R K R 6 144 G G D E I V M V I 10 19 L A L I L C V T K 5 181 D Q K G s C F I Q 10 170 S T V E G Y I A Y 5 198 R K G H I L E L L 10 176 I A Y R H D Q K G 5 217 E L V Q E G K A R 10 213 M A Ξ A E L V Q E 5 224 A R K T N P E I Q 10 30 E G S E E D L D A 4 1 M S N P R S L E Ξ 9 .33 . E E D L D A L E H 4 73 D S R E D P V S C 9 61 F Q Ξ E L E K F Q 4 89 H G R E G F L K G 9 70 Q A I D S R E D P 4 129 I Q A C R G E Q R 9 103 V K L E N L F E A 4 138 D P G E T V G G D 9 109 F E A L N N K N C 4 140 G E T V G G D E I 9 1 18 Q A L R A K P K V 4 204 E L L T E V T R R 9 127 Y I I Q A C R G E 4 207 T E V T R R M A E 9 130 Q A c R G E Q R D 4 232 Q S T L R K R L Y 9 164 D A L H V Y S T V 4 A L I L C V T K A 8 214 A E A E L V Q E G 4 22 I L C V T K A R E 8 2 S N P R S L E E E 3 60 Q F Q E E L E K F 8 5 R S L E E E K Y D 3 85 V L A H G R E G 8 .10 E K Y D S G A R 3 101 E M V K L E N L F 8 56 P T A E Q F Q E E 3 107 N L F E A L N N K 8 59' E Q F Q E E L E K 3 133 R G Ξ Q R D P G E 8 74 S R E D P V S C A 3 135 E Q R D .P G E T V 8 76 E D P V S C A F V 3 147 E I V M V I K D S 8 79 V S C A F V V L M 3 155 S P Q T I P T Y T 8 . 80 S c A F V V L M A 3 159 I P T Y T D A L H 8 124 P K V Y I I Q A C 3 201 H I L E L L T E V 8 143 V G G D E I V M V 3 202 I L E L L T E V T 8 166 L H V Y S T V E G 3 205 .L L T E V T R R M 8 188 I Q T L V D V F T 3 209 V T R R M A E A E 8 8 E E E K Y D M S G 2 18 R L A L I L C V T 7 32 S E E D L D A L E 2 42 M F R Q L R F E S 7 34 E D L D A L E H M 2 52 M K R D P T A E Q 7 46 L R F E S T M K R 2 75 R E D P V S C A F 7 66 E K F Q Q A I D S 2 120 L R A K P K V Y I 7 83 F V V L M A H G R 2 165 A L H V Y S T V E 7 90 G R E G F L K G E 2 168 V y S T V E G Y I 7 95 L K G E D G E M V 2 177 A Y R H D Q K G s 7 106 E N L F E A L N N 2 180 H D Q K G S C F I 7 132 C R G E Q R D P G 2 210 13P1 F11 v.l: HLA-B*08 iioniimers E13P1 F11 v.l: HLA-B*1510 nonamers Pos 1 2 3 4 5 6 7 8 9 score 40 E H M F R Q L R F 1 Portion of | 179 R H D Q K G S C F 17 SEQ ID 231 - I Q S T L R K R L 16 S R ,E D P V S C A 4 173 E G Y I A Y R H D 3 K N C Q A L R A K 4 178 Y H D Q K G S C 3 Y I I Q A C R G E 4 186 C F I Q T L V D V 3 I I Q A C R G E Q 4 189 Q T L V D V F T K 3 E Q R D P G E T V 4 191 • L V D V F T K R K 3 G E T V G G D E I 4 192 V D V F T K R K G 3 V G G D E I V M V 4 196 T K R K G H I L E 3 I V M V I K D S P 4 203 L E L L T E V T R 3 D S P Q T I P T Y 4 207 T E V T R R M A E 3 T y T D A L H V Y 4 208 E V T R R M A E A 3 T V E G Y I A Y R 4 213 M A E A E L V Q E 3 I Q T L V D V F T 4 217 E L V Q E G K A R 3 E L L T E V T R R 4 220 Q E G K A R K T N 3 R A E A E L V Q 4 223 K A R K T N P E I 3 A Ξ A E L V Q E G 4 226 K T N P E I Q S T 3 V Q E G K A R K T 4 229 P E I Q S T L R K 3 M S N P R S L E E ' 3 ■230 E I Q S T L R K R 3 E E E K Y D M S G 3 232 . Q s T L R K R L Y 3 E Y D M S G A R 3 7 L E E E K Y D M S 2 R L A L I L C V T 3 9 E E K Y D M S G A 2 L A L I L C V T K 3 24 C V T K A R E G S 2 L C V T K A R E G 3 32 S E E D L D A L E 2 T Κ· A R E G S E E 3 33 E E D L D A L E H 2 K A R E G S E E D 3 41 H M F R Q L R F E 2 E G S E E D L D A 3 49 E S T M K R D P T 2 M F R Q L R F E S 3 61 F Q E E L E K F Q 2 S T M K R D P T A 3 62 Q E E L E K F Q Q 2 T M K R D P T A E 3 63 E E L E K F Q Q A 2 M R D P T A E Q J 3 66 E K F Q Q A I D S 2 P T A E Q F Q E E 3 67 F Q Q A I D S R 2 E Q F Q E E L E K 3 68 F Q Q A I D S R E 2 V L M A H G R E G 3 70 Q A I D S R E D P 2 G R E G F L K G E 3 76 E D P V S C A F V 2 G F L K G E D G Ξ 3 77 D P V S C A F V V 2 E D G E M V K L E 3 80 S C A F V V L M A 2 D G E V K L E N 3 81 . C A F V V L M A H 2 V K L E N L F E A 3 82 A F V V L M A H G 2 E A L N N K N C Q 3 84 V V L M A H G R E 2 A L R A K P K V Y 3 89 H G R E G F L G 2 R A K P K V Y I I 3 91 R E G F L K G E D 2 I Q A C R G E Q R 3 109 F E A L N N K N C 2 Q A C R G E Q R D 3 114 N K N C Q A L R A 2 A C R G E Q R D P 3 118 Q A L R A K P K V 2 R G E Q R D P G E 3 123 K P K V Y I I Q A 2 G Ξ Q R D P G E T 3 124 P K V Y I I Q A C 2 R D P G E T V G G 3 126 V Y I I Q A C R G 2 E T V G G D E I V 3 132 C R G E Q R D P G 2 V I K D S P Q T I 3 138 D P G E T V G G D 2 I K D S P Q T I P 3 146 D E I V M V I K D 2 K D S P Q T I P T 3 147 E I V M V I K D S 2 212 T3P1F11 v. l: HLA-B*1510 nonamers Ω13Ρ1ΪΊ1 v.l: HLA-B*2705 noiiamer's Pos 1 2 3 4 5 6 7 8 9 score 197 K R K G H I L E L 29 46 . L R F E S T M K R 28 53 K R D P T A E Q F 25 28 A R E G s E E D L 24 4 P R S L E E E K Y 22 210 T R R M A E A E L 22 120 L R A K P K V Y I 21 97 G E D G E M V K L 19 17 A R L A L I L C V 18 59 E Q F Q E E L E K 18 67 K F Q Q A I D S R 18 107 N L F E A L N N K 18 125 K V Y I I. Q A C R 18 179 R H D Q K G S C F 18 '204 E L L T E V T R R 18 229 P E I Q S T L R K 18 75 R E D P V s C A F 17 100 G E M V K L E N L 17 218 L V Q E G K A R K 17 11 K Y D M S G A R L 16 44 R Q L R F E S T M 16 90 G R E G F L K G E 16 96 K G E D G E M V K 16 136 Q R D P G E T V G 16 171 T V E G Y I A Y R 16 183 K G S C F I Q T L 16 190 T L V D V F T K R 16 198 R K G H I L E L L 16 211 R R M A E A E L V 16 227 T N P E I Q S T L 16 19 L A L I L C V T 15 31 G S E E D L D A L 1 40 E H M F R Q L R F 15 57 T A E Q F Q E E L 15 60 Q F Q E E L E K F 15 · 101 E M V K L E N L F 15 115 K N C Q A L R A K 15 145 G D E I V M V I K 15 154 D S P Q T I P T Y 15 203 L E L L T E V T R 15 45 Q L R F E S T M K 14 81 C A F V V L M A H 14 121 R A K P K V Y I I 14 144 G G D E r V M V I 14 189 Q T L V D V F T 14 215 E A E L V Q E G K 14 217 E L V Q E G K A R 14 231 I Q S T L R K R L 14 233 S T L R K R L Y L 14 E K Y D M S G A R 13 13 D M S G A R L A L 13 S G A R L A L I L 13 21 L I L C V T. K A R 13 36 L D A L E H M F R 13 213 (213P1F11 v.l: HLA-B*2705 nonamers I213P1F11 v. l: HLA-B* 2705 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 74 s R E D P V S C A 13 151 V I K D S P Q T .1 8 83 F V V L M A H G R 13 184 G s c F I Q T L V 8 94 F L K G E D G E 13 201 H I L E L L T E V 8 104 K L E N L F E A L 13 226 K T N P E I Q S T 8 113 N N K N C Q A L R 13 27 K A R E G S E E D 7 170 S T V E G Y I A Y 13 66 . E K F Q Q A I D S 7 172 V E G Y I A Y R H 13 91 R E G F L K G E D 7 187 F I Q T L V D V F 13 106 E N L F E A L N N 7 223 K A R K T N P E I 13 123 K P K V Y I I Q A 7 228 N P E I Q S T L R 13 150 M V I K D S P Q T 7 230 E I Q s T L R K R 13 168 V Y s T V E G Y I 7 3 N P R s L E E E K 12 212 R M A E A E L V Q 7 6 S L E E E K Y D M 12 225 R K T N P E I Q s 7 33 E E D L D A L E H 12 16 G A R L A L I. L c 6 38 A L E H M F R Q L 12 47 R F E S T M K R D 6 43 F R Q L R F E s T 12 89 H G R E G F L K G 6 78 P V S C A F V V L 12 1 18 Q A L R A K P K V 6 86 L M A H. G R E G F 12 13 1 A C R G E Q R D P 6 87 M A H G R E G F L 12 141 E T V G G D E I V 6 88 A H G R E G F L 12 146 D E I V M V I K D 6 112 L N N K N C Q A L 12 152 I K D S P Q T I P 6 117 C Q A L R A K P 12 176 I A Y R H D Q K G 6 129 I Q A C R G E Q R 12 186 C F I Q T L V D V 6 140 G E T V G G D E I 12 219 V Q E G K A R K T 6 142 T V G G D E I V M 12 8 E E E K Y D M S G 5 178 Y R H D Q K G S C 12 22 I L C V T K A R E 5 191 L V D V F T K R K 12 29 R E G S Έ E D L O 5 193 D V F T K R K- G H 12 37 D A L E H M F R Q 5 205 L L T E V T R R M 12 50 S T M K R D P T A 5 34 E D •L D A L E H 11 63 E E L E K F Q Q A 5 D L D A L E H M F 11 68 F Q Q A I D s R E 5 119 A L R A K P K V Y 11 72 I D S R E D P V S 5 132 C R G E Q R D P G 11 92 E G F L K G E D G 5 159 I P T Y T D A L H 11 103 V K L E N L F E A 5 167 H V Y S T V E G Y 11 108 L F E A L N N K N 5 175 Y I A Y R H D Q K 11 122 A K P K V Y I I Q 5 180 H D Q K G S C F I 11 124 P K V Y I I Q A C 5 195 F T K R K G H I L 11 126 V Y I I Q A c R G 5 . 224 A R K T N P E I Q 11 143 V G G D E I V M V 5 39 . L Ξ H M F R Q L R 10 147 E I V M V I K D S 5 64 E L E K F Q Q A I 10 157 Q T I P T Y T D A 5 79 V S C A F V V L M 10 164 D A L H V Y s T V 5 158 T I P T Y T D A L 10 165 A L H V Y S T V E 5 161 T Y T D A L H V Y 10 174 G Y I A Y R H D Q 5 200 G H I L E L L T E 10 185 S C F I Q T L V D 5 232 Q S T L R K R L Y 10 188 I Q T L V D V F T 5 A L I L C V T K A 9 196 T K R K G H I L E 5 93 G F L K G E D G E 9 214 A E A E L V Q E G 5 194 V F T K R K G H I 9 216 A E L V Q E G K A 5 R S L E E E K Y D 8 221 E G .K A R K T N P 5 14 M S G A R L A L I 8 222 G K A R K T N P E 5 18 R L A L I L C V T 8 1 M S N P R S L E Ξ 4 133 R G E Q R D P G E 8 23 L c V T K A R E G 4 137 R D P G E T V G G 8 30 E G S E E D L D A 4 214 P13P1F11 v.l: HLA-B*2705 nonamers P13P1F11 v.l: HLA-B*2705 nonamers Pos 1 2 3 4 5 6 7 8 9 score 41 H M F R Q L R F E 4 42 M F R Q L R F E S 4 54 R D P T A E Q F Q 4 55 ' D P T A E Q F Q E 4 62 Q E E L E K F Q Q 4 82 A F V V L M A H G 4 84 V V L M A H G R E 4 99 D G E M V K L E N 4 102 M V K L E N . L F E 4 109 F " E A L N N K N C 4 1 10 E A L N N K N C Q 4 1 1 1 A L N N K N C Q A 4 114 N K N C Q A L R A 4 116 N c Q A L R A K P 4 127 Y I I Q A C R G E 4 130 Q A c R G E Q R D 4 134 G E Q R D P G E T 4 148 I V M- V I K D S P 4 149 V M V I K D S P Q 4 153 K D S P Q T I P T 4 160 P T Y T D A L H V . 4 163 T D A L H V Y S T 4 166 L H V Y S T V E G 4 192 V D V F T K R K G 4 199 K G H I L E L L T 4 202 I L E L L T E V T 4 213 M A E A E L V Q E 4 234 T L R K R L Y L Q 4 2 S N P R S L E E E 3 12 Y D M S G A R L A 3 26 T K A R E G S E Ξ 3 32 s E E D L D A L Ξ 3 52 M K R D P T A E Q 3 65 L E K F Q Q A I D 3 70 Q A I D s R E D P 3 73 D S R E D P V S C 3 77 D P V S C A F V V 3 80 S c A F V V L M A 3 155 S P Q T I P T Y T 3 156 P Q T I P T Y T D 3 177 A Y R H D Q K G S 3 181 D Q K G S C F I Q 3 220 Q E G K A R K T N 3 7 L E E E K Y D M S 2 24 C V T K A R E G S 2 51 T M K R D P T A E 2 56 • P T A E Q F Q E E 2 61 F Q E E L E K F Q 2 69 Q Q A I D S R E D 2 76 E D P V S C A F V 2 98 E D G E M V K L Ξ 2 105 L E N L F E A L N 2 128 I I Q A C R G E Q 2 135 E Q R D P G E T V 2 215 213P1F11 v.l : HLA-B*2709 nonamers 213P1F11 v.l: HLA-B*27()9 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 223 K A R K T N P E I 11 145 G D E I V M V I K 4 224 A R K T N P E I Q 11 150 M V I K D S P Q T 4 227 T N P E I Q S T L 11 167 H V Y s T V E G Y 4 4 P R S L E E E K Y 10 172 V E G Y I A Y R H 4 34 E D L D A L E H M 10 174 G Y I A Y R H D Q 4 40 E H M F R Q L R F 10 189 Q T L V D V F T K 4 43 F R Q L R F E S T 10 204 E L L T E V T R R 4 57 T A E Q F Q E E L 10 16 G A R L A L I L C 3 71 A I D S R E D P V 10 19 L A L I L C V T K 3 79 V S C A F V V L M 10 20 A L I L C V T K A 3 87 M A H G R E G F L 10 33 E E D L D A L E H 3 112 L N N K N C Q A L 10 37 D A L E H M F R Q 3 132 C R G E Q R D P G 10 59 E Q F Q E E L E K 3 158 T I P T Y T D A L 10 63 E E L E K F Q Q A 3 164 D A L H V Y S T V 10 66 E K F Q Q A I D ■S 3 178 Y R H D Q K G s C 10 84 V V L M A H G R E 3 186 C F I Q T L V D V 10 107 N L F E A L N N K . 3 195 F T K R K G H I L 10 114 N K N C Q A L R A 3 201 H I L E L L T E V 10 123 K P K V Y I I Q A 3 205 L L T E V T R R M 10 126 V Y I I Q A c R G 3 6 S L E E E K Y D M 9 153 ■ K D s P Q T I P T 3 95 L K G E D G E M V 9 176 I A Y R H D Q K G 3 101 E M V K L E N L F 9 185 s C F I Q T L V D 3 141 E T V G G D E I V 9 188 I Q T L V D V F T 3 142 T V G G D E I V M 9 199 K G H I L E L L T 3 143 V G G D E I V M V 9 203 L E L L T E V T R 3 187 F I Q T L V D V F 9 216 A E L V Q E G K A 3 194 V F T K R K G H I 9 222 G K A R K T N P E •3 14 M S G A R L A L I 8 226 K T N P E I Q S T 3 D L D A L E H M F 8 229 P E Γ Q S T L R K 3 60 Q F Q E E L E K F 8 1 M S N P R S L E E 2 64 E L E K F Q Q A I 8 10 E K Y D M S G A R 2 76 E D P V S C A F V 8 12 Y D M S G A R L A 2 86 L M A H G R E G F 8 21 L I L C V T K A R 2 94 F L K G E D G E M 8 22 I L C V T K A R E 2 135 E Q R D P G E T V 8 23 L C V T K A R E G 2 151 V I K D S P Q T I 8 27 K A R E G S E E D 2 168 V Y S T V E G Y I 8 41 H M F R Q L R F E 2 180 H D Q K G S C F I 8 55 D P T A E Q F Q E 2 R S L E E E K Y D 6 67 K F Q Q A I D S R 2 47 R F E S T M K R D 6 68 F Q Q A I D S R E 2 225 R K T N P E I Q S 6 72 I D S R E D P V S 2 18 R L A L I L c V T 5 73 D S R E D P V S C 2 29 R E G S E E D L D 5 80 S C A F V V L M A . 2 93 G F L K G E D G E ' 5 81 C A F V V L M A H 2 106 E N L F E A L. N N 5 82 A F V V L M A H G 2 125 K V Y I I Q A C R 5 83 F V V L M A H G R 2 133 R G E Q R D P G E 5 92 E G F L K G E D G 2 137 R D P G E T V G G 5 96 K G E D G E M V K 2 200 G H I L E L L T E 5 103 V K L E N L F E A 2 212 R M A E A E L V Q 5- 110 E' A L N N K N C Q 2 54 R D P T A E Q F Q 4 111 A L N N K N C Q A 2 91 R E G F L K G E D 4 115 K N C Q A L R A K 2 134 G E Q R D P G E T 4 124 P K V Y I I Q A ■C 2 216 213P1F11 v.l: HLA-B*2709 nonamers E13P1F11 v.l: HLA-B*2709 nonamers Pos 1 2 3 4 5 6 7 8 9 score 129 I Q A C R G E Q R 146 D E I V M V I K D 148 I V M V I K D S P 152 I K D S P Q T I P 156 P Q T I P T Y T D 157 Q T I P T Y T D A 159 I P T Y T D A L H 163 T D A L H V Y S T 166 L H V Y S T V E G 169 Y S T V E G Y I A 173 E G Y I A Y R H D 177 A Y R H D Q.K G S E13P1F11 v.l: HLA-B*4402 nomimers 182 Q K G S C F I Q T 193 D V F T K R K G H Portion of 213 M A E A E L V Q E SEQED 214 A E A E L V Q E G I O: 3; each| N P R S L E E E K start position is E E K Y D. M S G A specified, 24 C V T K A R E G S the length E G S E E D L D A of each 49 E S T M K R D P T peptide is 9 50 S T M K R D P T A amino 51 T M K R D P T A E acids, the A E Q F Q E E L E jend position] 58 for each 62 Q E E L E K F Q Q peptide is 69 Q Q A I D S R E D the start 70 Q A I D S R E D P position 88 A H G R E G F L K plus eight 89 H G R E G. F L K G 98 E D G E M V K L E 99 D G E M V K L E N 102 M V K L E N L F E 109 F E A L N N K N C 117 C Q A L R A K P K 119 A L R A K P K V Y 122 A K P K V Y I I Q 127 Y I I Q A C R G E 128 I I Q A C R G E Q 130 Q A C R G E Q R D 131 A C R G E Q R D P 138 D P G E T V G G D 147 E I V M V I K D S 149 V M V I K D S P Q 154 D S P Q T I P T Y 155 S P Q T I P T Y T 161 T Y T D A L H V Y 162 Y T D A L H V Y S 165 A L H V Y S T V E 170 .S V E G Y I A Y 175 Y I A .Y H D Q K 190 T L V D V F T K R 191 L V D V F T K R K 192 V D V F T K R K.G 202 I L E L L T E V T 217 Q13P1F11 v.l: HLA-B*4402 nonamers Q13P1F11 v.l: HLA-B*4402 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 227 T N P E I Q S T L 13 70 Q A I D S R E D P 5 8 E E E K Y D M S G 12 71 . A I D S R E D P V 5 9 E E K Y D M S G A 12 81 C A F V V L M A H 5 1 1 K Y D M S G A R L 12 88 A H G. R E G F L K 5 D L D A L E H M F 12 92 E G F L K G E D G 5 39 L E. H M F R Q L R 12 106 E N L F E A L N N 5 60 Q F Q E E L E K F 12 107 N L F E A L N N K 5 62 Q E E L E K F Q Q 12 1 1 1 A L N N K N C Q A 5 64 E L E F Q Q A I 12 115 K N C Q A L R A K 5 ■ 87 M A H G R E G F L 12 117 C Q A L R A K P K 5 105 L E N L F E A L N 12 122 A K P K V Y I I Q 5 121 R A K P K V Y I I 12 13 1 A C R G E Q R D P 5 134 G E Q R D P G E T 12 136 Q R D P G E T V G 5 144 G G D E I V M V I 12 143 V G G D E I V M V 5 172 V E G Y I A Y R H 12 174 G Y I A Y R H D Q 5 195 F T K R K G H I L 12 177 A Y R H D Q K G s 5 14 M S G A R L A L I 1 1 186 C F I Q T L V D V 5 29 R E G S E E D L D 1 1 204 E L L T E V T R R 5 86 L M A H G R E G F 11 230 E I Q S T L R K R 5 151 V I K D S P Q T I 1 1 2 S N P R S L E E E 4 167 H V Y S T V E G Y 1 1 12 Y D M S G A R L A 4 179 R H D Q K G S C F 1 1 16 G A R L A L I L C 4 7 L E E E K Y D M S 10 18 R L A L I L C V T 4 57 T A E Q F Q E E L 10 30 E G S E E D L D A 4 65 L E K F. Q Q A I D JO 46 L R F E S T M K R 4 91 R E G F L K G E D 10 50 S T M K R D P T A 4 168 V Y S T V E G Y I 10 51 T M K R D P T A E 4 210 T R R M A E A E L 10 67 K F Q Q A I D S R 4 A L I L C V T K A 9 90 G R E G F L K G E 4 120 L R A K P K V Y I 9 126 V Y I I Q A C R G 4 194 V F T K R K G H I 9 127 Y I I Q A C R G E 4 223 K A R K T N P E I 9 135 E Q R D P G E T V 4 17 A R L A. L I L C V 8 137 R D P G E T V G G 4 180 ,H D Q K G s C F I 8 150 M V I K D S P Q T 4 147 E I V M V I K D s 7 155 S P Q T I P T Y T 4 153 K D S P Q T I P T 7 165 A L H V Y S T V E 4 200 G H I L E L L T E 7 171 T V E G Y I A Y R 4 226 K T N P E I Q S T 7 191 L V D V F T K R K 4 21 L I L C V T K A R 6 208 E V T R R M A E A 4 66 E K F Q Q A I D S 6 209 V T R R M A E A E 4 98 E D G E M V K L E 6 219 V Q E G K A R K T 4 1 10 E A L N N K N C Q 6 5 R s L E E E K Y D 3 123 K P K V Y I I Q A 6 19 L A L I L C V T 3 124 P K V Y I I Q A C 6 23 L C V T K A R E G 3 157 Q T I P T Y T D A 6 43 F R Q L R F E S T 3 185 S C F I Q T L V D 6 44 R Q L R F E S T M 3 193 D V F T K R K G H 6 49 E S T M K R D P T 3 217 E L V Q E G K A R 6 72 I D S R E D P V S 3 224 A R K T N P E I Q 6 74 S R E D P V S C A 3 1 M S N P R S L E E 5 76 E D P V S C A F V 3 E K Y D M S G A R 5 79 V S C A F V V L M 3 34 E D L D A L E H M 5 80 S C A F V V L M A 3 41 H M F R Q L R F E 5 82 A F V V L M A H G 3 59 E Q F Q E E L E K 5 83 F V V L M A H G R ' 3 218 E13P1F11 v.l : HLA-B*4402 nonamers 213P1F11 v.l: HLA-B 402 nonamers Pos 1 2 3 4 5 6 7 8 9 score 89 H G R E G F L K G 3 96 K G E D G E M V K 3 103 V K L E N L F E A 3 113 N N K N C Q A L R 3 114 N K N C Q A L R A 3 116 N c Q A L R A K P 3 118 Q A L R A K P K V 3 141 E T V G G D E I V 3 142 T V G G D E I V M 3 160 P T Y T D A L H V 3 173 E G Y I A Y R H D 3 182 Q K G s C F I Q T 3 189 Q T L V D V F T K 3 190 T L V D V F T K R 3 199 G H I L E L L T 3 202 I L E L L T E V T 3 211 R R M A E A E L V 3 213 M A E A E L V Q E 3 215 E A E L V Q E G K 3 221 E G K A R K T N P 3 222 G K A R K T N P E 3 225 R K T N P E I Q S 3 234 T L R K R L Y L Q 3 47 R F E s T M K R D 2 52 M K R D P T. A E Q 2 54 D P T A E Q F Q 2 61 F Q E E L E K F Q 2 73 D • Q T F R L K E E Q 5 43 R G S S V H Q K L 5 61 V F G G G V G D I 5 66 V G D I V G R D L 5 13 V Q P E K R T G L 4 21 L R D E N G E C G 4 28 C G Q T F R L K E 4 49 Q K L V N D P R E 4 55 P R E T Q E V F G 4 56 R E T Q E V F G G 4 58 T Q E V F G G G V 4 78 F R N S E T S A S 4 85 A S E E E K Y D M 4 86 S E E E K Y D M S 4 4 C Q E Y. D K S L S 3 37 E Q G R A F R G S 3 53 N D P R E T. Q E V 3 57 E T Q E V F G G G 3 232 233 F11 V.4: HLA-B* 08 nonamers E13P1F11 v.4: HLA-B*08 nonamers 1 2 3 4 5 6 7 Θ 9 score E N 6 E C G Q T F 10 E E Q G R A F R G 10 R G s S V H Q K L 10 bl3PlFll v.4: HLA-B*1510 nonamers S L s V Q P E K R 9 Pos 1 2 3 4 5 6 7 8 9 score R T G L R D E N G 9 26 G E C G Q T F R L 14 Portion of L K E E Q G R A F 9 13 V Q P E K R T G L 12 SEQ ID G R A F R G S S V 9 47 V H Q K L V N D P 12 NO: 9, eachl start V N D P R E T Q E 9 66 V G D I V G R D L 12 position is S A S E E E K Y D 9 3 K C Q E Y D K 3 L 11 specified, E K R T G L R , D E 8 34 L K E E Q G R A F 11 the length V F G G G V G D I 8 43 R G S S V H Q K L 11 of each S F R N S E T S A 8 24 E N G E c G Q T F 9 peptide is 9 A F R G S S V H Q 7 54 D P R E T Q E V F 9 amino K L V N D P R E T 7 85 A S E E E K Y D M 9 acids, the ' Q G R A F R G S S 6 70 V G R D L S I S F 7 |end position| for each D L S I S F R N S 6 12 S V Q P E K R T G 6 peptide is ' R A F R G S S V H 4 49 Q K L V N D P R E 5 the start E T Q E V F G G G 4 60 E V F G G G V G 0 5 position G S S V H Q K L V 3 63 G G G V G D Ί V G 5 plus eight E V F G G G V G D 3 64 G G V G D I V G R 5 G G V G D I V G R 3 65 G V G D I V G R D 5 G R D . L S I S F R 3 6 E Y D K S L S V Q 4 S E T S A S E E E 3 11 L S V Q P E K R T 4 E T S A S E E E 3 16 E K R T G L R D E 4 E Y D K S L S V Q 2 27 E C G Q T F R L K 4 D K S L S V Q P E 2 35 K E E Q G R A F R 4 S V Q P E K R T G 2 36 E Ξ Q G R A F R G 4 L R D E N G E C G 2 50 K L V N D P R E T 4 E C G Q. T F R L K 2 51 L V N D P R E T Q 4 .

E Q G R A F R G S 2 67 G D I V G R D L s 4 F R G S S V H Q K 2 76 I S F R N S E T s 4 S S V H Q K L V N 2 7 Y D K S L S V Q P 3 V H Q K L V N D P 2 10 S L S V Q P E K R 3 G G G V G D I V G 2 17 K R T G L R D E N 3 G V G D I V G R D 2 20 G L R D E N G E C 3 G D I V G R D L S 2 33 R L K E E Q G R A 3 F R N S E T S A S 2 37 E Q G R A F R G S 3 R N S E T S A S E 2 40 R A F R G S S V H 3 A S E E E K Y D M 2 41 A F R G S S y H Q 3 G K C Q E Y D K S 1 44' G S S V H Q K L V 3 C Q E Y D K S L S 1 45 S S V H Q K L V N 3 K S L S V Q P E K 1 46 'S V H Q K L V N D 3 L S V Q P E K R T 1 55 P R E T Q E V F G 3 K R T G L R D ■E N 1 58 T Q E V F G G G V 3 Q T F R L K E E Q 1 59 Q E V F G G G V G 3 F R L K Ξ E Q G R 1 61 V F G G G V G D I 3 Q K L V N D P R E 1 73 D L S I S F R N s 3 N D P R E T Q E V 1 82 E T S A S E E E K 3 P R E T Q E V F G 1 2 G K C Q E Y D K s 2 R E T Q Ξ V F G G 1 4 C Q E Y D K S L s 2 T Q Ξ V F G G G V 1 8 D K S L S V Q P E 2 F G G G V G D I V 1 9 K s L S V Q P E K 2 I V G R D L S I S 1 14 Q P E K R T G L R 2 I S F R N S E T S 1 15 P E K R T G L R D 2 234 B13P1F11 v.4: HLA-B* 1510 nonamers B13P1F11 v.4: HLA-B*2705 nonamers Pos 1 2 3 4 5 6 7 8 9 score 14 Q P E K R T G L R 12 24 . E N G E C G Q T F 12 27 E C G Q T F R L K 12 68 D I V G R D L S I 12 78 F R N S E T S A s 12 82 E T S A . S E E E K 12 1 M G K C Q E Y D K 1 1 21 L R D E N G E C G 1 1 34 L K E E Q G R A F 11 55 P. R E T Q E V F G 11 61 V F G G G V G D I 11 66 V G D I V G R D L 11 83 T S A S E E E K Y 1 1 33 R L K E E Q G R A 10 72 R D L S I S F R N 10 2 - G K C Q E Y D K S 8 18 R T G L R D E N G 8 56 R E T Q E V F G G 8 60 E V F G G G V G D 8 63 G G G V G D I V G 8 79 R N Ξ E T S A S E 8 46 S V H Q K L V N D 7 65 G V G D I V G R D 7 67 G D I V G R D L S 7 Q E Y D K S L S V 6 6 E Y D K S L S V Q 6 1 1 L s V. Q P E K R T 6 G L R D E N G E c 6 22 R D E N G E C G Q 6 Q T F R L K E E Q 6 41 A F R G S S V H Q 6 49 Q K L V N D P R E 6 76 I s F R. N S E T S 6 29 G. Q T F R L K E E 5 36 E Ξ Q G R A F R G 5 44 G S S V H Q K L V 5 47 V H Q K L V N D P 5 . 7 Y D K S L S V Q P 4 12 S V Q P E K R T G 4 23 D E N G E C G Q T 4 45 S S V H Q K L V N 4 50 K L V N D P R E T 4 52 V N D P R E T Q E 4 59 Q E V F G G G V G 4 74 L S I S F R N S E 4 77 S F R N S E T S A 4 80 N S E T S A S E E 4 8 D K S L S V Q P E 3 P Ξ K R T G L R D 3 16 E K R T G L R D E 3 19 T G L R D E N G E 3 28 c G Q T F R L K E 3 3 1 T F R L K E E Q G 3 62 F G G G V G D I V 3 235 B13P1F11 v.4: HLA-B*2705 nonamers B13P1F11 v.4: HLA-B*2709 nonamers Pos 1 2 3 4 5 6 7 8 9 score 7 Y D K S L S V Q P 3 19 T G L R D E N G E 3 G L R D E N G E C 3 . 60 E V F G G G V G D 3 63 G G G V G D I V G 3 76 I S F R N S E T S 3 11 L S V Q P E K R T■ 2 P E K R T G L R D 2 Q T F R L K E E Q 2 K E E Q G R A F R 2 36 E E Q G R A F R G 2 41 A F R G S S V H Q 2 45 S S V H Q K L V N 2 46 S V H Q K L V N D 2 52 V N D P R E T Q E 2 69 I V G R D L S I S 2 74 L S I S F R N S E 2 81 S E T S A S E E E 2 1 M G K C Q E Y D K 1 4 C Q E Y D K S L S 1 8 D K S L S V Q P E 1 12 S V Q P, E K R T G 1 23 D E N G E c G Q T 1 28 e G Q T F R L K E 1 31 T F R L K E E Q G 1 47 V H Q K L V N D P 1 59 Q E V F G G G V G 1 73 D L S I S F R N S 1 80 N S E T S A S E E 1 83 T S A S E E E K Y 1 213P1F11 v.4: HLA-B* 4402 nonamers Pos 1 2 3 4 5 6 7 8 9 score 26 G Ξ C G Q T F R L 22 Portion of 36 E E Q G R A F R G 15 SEQ ID 3 K C Q E Y D K S L 13 NO: 9; each E N G E C G start 24 Q T F 13 position is 34 L K E E Q G R A F 13 specified, Q E Y D K S L S V 12 the leneth 13 V Q P E K R T G L 12 of each P E K R T G L R D 12 peptide is 9 23 D E N G E C G Q T 12 amino K E E Q G R A F R 12 acids, the 43 R G S S V H K L 12 end position Q for each 66 V G D I V G R D L 12 peptide is 70 V G R D L S I S F 12 the start 54 D P R E T Q E V F 11 position 56 R E T Q E V F G G 11 plus eight 59 Q E V F G G G V G 11 68 D I V G R D L S I 11 81 S E T S A S E E E 11 83 T S A S E E E K Y 11 86 S E E E K Y D M S 11 61 V F G G G V G D I 10 236 ai3PlFll v.4: HLA-B*4402 nonamers G13P1F11 v.4: HLA-B*4402 nonamers Pos 1 2 3 4 5 6 7 8 9 score 12 s V Q P E K R T G 6 52 V N D P R E T Q E 6 60 E V F G G G V G D 6 6 E Y D K S L S V Q 5 16 E K R T G L R D E 5 ' 27 E c G Q T F R L K 5 37 E Q G R A F R G S 5 41 A F R G S S V H Q 5 . 64 G G V G D I V G R 5 67 G D I V G R D L S 5 P13P1F11 v.4: HLA-B*5101 nonamers 71 G R D L S I S F R 5 Pos 1 2 3 4 5 6 7 8 9 score 74 L S I S F R N S E 5 54 D P R E T Q E V F ' 20 Portion of 76 I S F , R N S E T S 5 62 F G G G V G D I V 18 SEQ ID S L S V Q P E K R 4 43 R G S S V H Q K L 16 [NO: 9; eachl C G R L K 4 start 28 Q T F 66 V G D I V G R D L 16 position is 29 G Q T F R L K E E 4 68 D I V G R D L S I 16 specified, 40 R A F R G S S V H 4 5 Q E Y D K S L S V 15 the length 44 G S S V H Q K L V 4 40 R A F R G S S V H 14 of each 50 K L V N D P R E T 4 61 V F G G G V G D I 14 peptide is 9 53 N D P R E T Q E V 4 84 S A S E E E K Y D 13 amino 84 S A S E E E K Y D 4 13 V Q P E K R T G L 12 acids, the 8 D K S L S V Q P E 3 ]end position| 14 Q P E K R T G L R 11 for each 14 Q P E K R T G L R 3 3 K C Q E Y D K S L 10 peptide is 17 K R T G L R D E N 3 19 T G L R D E N G E 10 the start 19 T G L R D E N G E 3 28 C G Q T F R L K E 10 position Q T F R L K E E Q 3 53 N D P R E T Q E V 10 plus eight 42 F R 'G S S V H Q K 3 63 G G G V G D I V G 10 45 S S V H Q K L V N 3 44 G S S V H Q K L V 9 46 S V H Q K L V N D 3 ■ 58 ■ T Q E V F G G G V 9 51 L V N D P R E T Q 3 64 G G V G D I V G R 9 57 E T Q E V F G G G 3 70 V G R D L S I S F ' 9 65 G V G D I V G R D 3 25 N G E C G Q T F R 8 73 D L S I S F R N S. 3 26 G E C G Q T F R L 8 75 S I S F R N S E T 3 39 G R A F R G S S V 8 78 F R N S E T S A S 3 1 M G K C Q E Y D K 7 80 N S E T S A S E E 3 8 D K S L S V Q P E 7 82 E T S A S E E E K 3 38 Q G R A F R G S S 7 85 A S E E E K Y D M 3 73 D L S I S F R N S 7 7 Y D K S L S V Q P ' 2 6 E Y D K S L S V Q 6 9 K S L S V Q P E K 2 23 D E N G E C G Q T 5 11 L S V Q P E K R T 2 34 L K E E Q G R A F 5 18 R T G L R D E N G 2 47 . V H Q K L V N D P 5 21 L R D E N G E C G 2 51 L V N D P R E T Q 5 31 T F R L K E E Q G 2 57 E T Q E V F G G G 5 47 V H Q K L V N D P 2 76 I S F R N S E T S 5 49 Q K L V N D P R E 2 9 K S L S V Q P E 4 55 P R E T Q E V F G 2 11 L S V Q P E K R T 4 63 G G G V G D I V G 2 21 L R D E N G E C G 4 69 I V G R D L S I S 2 24 E N G E C G Q T F 4 72 R D L S I S F R N 2 32 F R L K E E Q G R 4 77 S F R N S E T S A 2 41 A F R G S S V H Q 4 2 G K C Q E Y D K S 1 42 F R G S S V H Q K 4 4 C Q E Y D K S L S 1 45 S S V H Q K L V N 4 G L R D E N G E C 1 65 G V G D I V G R D 4 237 I213P1F11 v.4: HLA-B*5101 nonamers Q13P1F11 v.4: RT1.A1 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 80 N S E T S A S E E . 4 ' 76 ■ I S F R N S E T S 18 Portion of | 7 Y D K S L S V Q P 3 83 T S A S E E E K Y 18 SEQID S L S V Q P E K R 3 60 E V F G G G V G D 17 [NO: 9; each| S V P E K R T G start 12 Q 3 85 A S' E E E K Y D M 17 position is j P E K R T G L R D 3 40 R A F R G S S V H 16 specified, 16 E K R T G L R D E 3 9 K S L S V Q P E K 14 the length 27 E c G Q T F R L K 3 30 Q T F R L K E E Q 13 of each 33 R L K E E Q G R A 3 24 E N G E C G Q T F 12 peptide is 9 36 E E Q G R A F R G 3 54 D P R E T Q E V F 12 amino 46 S V H Q K L V N D 3 12 S V Q P E K R T G 11 acids, the 48 H Q K L V N D P R 3 34 L K E E Q G R A F 11 |end position for each 49 Q K L V N D P R E 3 5 Q E Y D K S L S V 10 peptide is 59 Q Ξ V F G G G V G 3 70 V G R D L S I S F 10 the start 60 E V F G G G V G D 3 51 L V N D P R E T Q 9 position 69 I V G R D L S I S 3 84 S A S E E E K Y D 9 plus eight 72 R D L S I S F R N 3 3 K C Q E Y D K s L 8 74 L S I S F R N S Ξ 3 13 V Q P E K R T G L 8 79 R N S E T S A S E 3 32 F R L K E E Q G R 8 83 T S A S E E E K Y 3 45 S S V H Q K L V N 8 86 S E E E K Y D M s 3 46 S V H Q K L V N D 8 2 G K C Q E Y D K s 2 74 L S I S F R N S Ξ 8 G L R D E N G E c 2 11 L S V Q P E K R T 7 29 G Q T F R L K E E 2 19 T G L R D E N G E 7 31 T F R L K E E Q G 2 26 G E C G Q T F' R L 7 37 E Q G R A F R G S •2 66 V G D I V G R D L 7 50 K L V N D P R E T 2 69 I V G R D L S I S •7 52 V N D P R E T Q E 2 80 N S E T S A S E E 7 55 P R E T Q E V F G 2 43 R G S S V H Q K L 6 56 R E T Q E V F G G 2 44 G S S V H Q K L V 6 71 G R D L S I S F R 2 49 Q K L V N D P R E 6 77 S F R N S E T S A 2 65 G V G D I V G R D 6 82 E T S A S E E E K 2 72 R D L S I S F R N 6 85 A S E E E K Y D M 2 17 K R T G L R D E N 5 17 K R T G L R D E N 1 2 G K C Q E Y D K S 3 Q T F R L K E E Q 1 35 K E E Q G R A F R 3 75 s I S F R N S E T 1 50 K L V N D P R E T 3 78 F R N S E T S A S 1 81 S E T S A S E E E 3 86 s E E E K Y D M S 3 4 c Q E Y D K S L S 2 s L S V Q P E K R 2 18 R T G L R D E N G 2 G L R D E N G E C 2 21 L R D E N G E C G 2 23 D E N G E C G Q T 2 28 C G Q T F R L K E 2 31 T F R L K E E Q G 2 33 R L K E E Q G R A 2 38 Q G R A F R G S S 2 41 A F R G S S V H Q 2 47 V H Q K L V N D P 2 48 H Q K L V N D P R 2 53 N D P R E T Q E V 2 56 R E T Q E V F G G 2 61. V F G G G V G D I 2 , 238 P13P1F11 v.4: RT1.A1 nonamers Q13P1F11 V.5: HLA-A26 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 62 F G G G V G D I V 2 5 A L I L R V T K A 15 Portion of 64 G G V G D I V G R 2 3 R L A L I L R V T 13 67 G D I V G R D L S 2 9 R V T K A R E G S 12 75 S I s F R N S E T 2 6 L I L R V T K A R 11 82 E T s A S E E E 2 7 I L R V T K A R E 10 1 M G K C Q E Y D K 1 2 A R L A L I L R V 6 6 E Y D K S L S V Q 1 I G A R L A L I L R 4 8 D K S L S V Q P E 1 4 L A L I L R V T K 2 14 Q P E K R T G L R 1 8 L R V T K A R E G 2 16 E K R T G L R D E TABLE XIXA, part 5: MHC Class I nonamer analysis of 213P1F1 1 v.5 (aa 1-242) B13P1F11 v.5: HLA-A*0201 nonamers Pos 1 2 3 4 5 6 7 8 9 score A L I L R V T K A 24 Portion of 2 A R L A L I L R V 20 SEQ ID R L A L I L R V T 20 NO: 1 1; 3 each start 6 L I. L R V T K A R 14 position is 7 I L R V T K A R E 14 specified, 4 L A L I L R V T K 13 the length 1 G A R L A L I L R 9 of each 8 L R V T K A R E G 5 peptide is 9 9 R V T K A R E G S 4 amino acids, the end position for each peptide is the start position plus eight 213P1F11 v.5: HLA-A1 nonamers Pos 1 2 3 4 5 6 7 8 9 score 2 A R L A L I L R V 7 Portion of bl3PlFll v.5: HLA-B*0702 nonamers SEQ ID Pos 1 2 3 4 5 6 7 8 9 score NO: 11; 2 , A R L A L I L R V 11 Portion of each start SEQ ID position is NO: 1 1 ; specified, each start . the length position is of each specified, peptide is 9 the length I G A R L A L I L R 6 amino of each A L I L R V T K A 5 acids, the peptide is 3 R L A L I L R V T 2 end 5 A L I L R V T K A 10 9 amino 6 L I L R V T K A R 2 position fo 3 R L A L I L R V T 9 acids, the 4 L A L I L R V T K 1 reach 7 I L R V T K A R Ξ 4 end 7 I L R V T K A R E 1 peptide is 4 L A L I L R V T K 3 position 9 R V T K A R E G S 1 the start 9 R V T K A R E G s 3 for each position 1 G A R L A L I L R 2 peptide is plus eight 6 L I L R V T K A R 2 the start position plus eight 239 P13P1F11 v.5: HLA-B*08 nonamers E13P1F11 v.5: HLA-B*2709 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score A L I L R V T K A 16 Portion of 2 A R L A L I L R V 23 Portion of 7 I L R V T K A R E 14 SEQ ID 8 , L R V T K A R E G 12 1 G A R L A L I L R 12 NO: 11 ; 3 R L- A L I L R V T 5 each start 8 L R V T K A R E 6 11 9 R V T K A R E G S 5 position is 3 R L A L i L R V T 7 G A R L A L I L R specified, 1 3 6 L I L R V T K A R 6 the length 4 L A L I L R V T K 3 4 L A L I L R V T K 5 of each 5 A L I L R V T K A 3 2 A R L A L I L R V 1 peptide is 9 6 L I L R V T K A R 2 amino 7 I L R V T K A R E 2 acids, the end position for each peptide is the start position plus eight 213P1F11 v.5: HLA-B*1510 nonamers Q13P1F11 v.5: HLA-B 402 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 7 I L R V T K A R E 6 Portion of 5 A L I L R V T K A 10 Portion of 3 R L A L L L R V T 5 SEQ ID 2 A R L A L I L R V 8 SEQ ID 4 L A L I L R V T K 4 NO: 11 ; 6 L I L R V T K A R 7 NO: 11 ; each start each start 2 A R L A L I L R V 3 3 R L A L I L R position is position is 8 L R V T K A R E G 3 specified, specified, 9 R V T K A R E G S 2 the length the length 1 G A R L A L I L R 1 of each of each A L I L R V T K A 1 peptide is 9 peptide is 9 6 L I L R V T K A R 1 amino amino acids, the acids, the end position end position for each for each peptide is peptide is the start the start position position plus eight plus eight Q13P1F1 1 v.5: HLA-B*2705 nonamers Pos 1 2 3 4 5 6 7 8 9 score TABLE XIXA, part 6: MHC Class I nonamer 2 A R L A L I L R V 20 Portion of analysis of 213P1F11 v.6 (aa 1-242) 1 G A R L A L I L R 16 SEQ ID bl3PlFll v.6: HLA-A*0201 nonamers 4 L A L I L R V T K 15 NO: 11 ; Pos 1 2 3 4 5 6 7 8 9 score each start 8 L R V T K A R E G 14 1 K L E N L F E A M 16 Portion of position is 6 L I L R V T K A R 13 8 ■ A M N N K N C Q A 16 SEQ ID · specified, A L I L R V T K A 9 4 N L F E A M N N K 14 NO: 13; the length each start 3 R L A L I L R V T 8 of each 9 M N N K N C Q A L 12 position is 9 R V T K A R E G S 6 peptide is 9 2 L E N L F E A M N 6 specified, 7 I L R V T K A R E 5 amino 5 L F E A M N N K N 4 the length acids, the 7 E A M N N K N C Q 3 of each end position 6 F E A M N N K N C 2 peptide is 9 for each 3 E N L F E A M N N -2 amino peptide is acids, the the start end position position for each plus eight peptide is the start position plus eight 240 P13P1F11 v.6: HLA-B*0702 nonamers P13P1F11 v.6: HLA-Al nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 Θ 9 score 9 M N N K N C Q A L 12 Portion of 1 K L E N L F E A 14 Portion of 1 K L E N L F E A M 9 ■ L F E A M N N K N 13 SEQ 1D 8 A M N N K N C Q A 8 6 F E A M N N K N C 2 NO: 13; 7 E A M N N K N C Q 2 C each start 8 A M N N K N Q A 2 2 L E N L F E A M N 1 position is 2 L E N L F E A M N 1 specified, 3 E N L F E A M N 1 4 N L F E A M N N K 1 the length 5 L F E A M N N K N 1 of each 6 F E A M N N K N C 1 peptide is 9 amino acids, the end position for each peptide is the start position plus eight bl3PlFll v.6: HLA-B*1510 nonamers E13P1F11 v.6: HLA-A3 nonamers Pos 1 2 3 4 5 6 7 8 9 score Pos 1 2 3 4 5 6 7 8 9 score 9 M N N K N C Q A L 13 Portion of 4 N L F E A M N N 21 Portion of 1 K L E N L F E A M 8 SEQ ID 1 K L E N L F E A M 15 SEQ ID 7 E A M N N K N C Q 3 NO: 13; C each start 8 A M N N K N Q A 8 NO: .13; 4 N L F E A M N N K 2 each start position is 3 E N L F E A M N N 6 6 F E A M N N K N C 2 position is specified, 2 - L E N L F E A M N 5 specified, 3 E N L F E A M N N 1 the length L F E A M N N K N 2 the length 5 L F E A M N N K N 1 of each 6 F E A M N N K N C 1 of each peptide is 9 7 E A M N N K N C Q 1 peptide is 9 amino 9 M N N K N C Q A L .1 amino acids, the acids, the end position end position for each for each peptide is peptide is the start the start position position plus eight plus eight 241 213P1F11 v.6: HLA-B*2705 nonamers TABLE XIXB: MHC Class I Analysis of Pos 1 2 3 4 5 6 7 8 9 score 213P1F11 (decamers). Listed are scores which 4 N L F E A M N N K 17 Portion of correlate with the ligation stiengtli to a defined HLA 9 M N N K N C Q A L 12 SEQ ID type for a sequence of amino acids. The algoritluns 1 K L E N L F E A M 11 NO: 13; used are based on tire book "MHC Ligands and E N L F E A .M N each start 3 7 Peptide Motifs" by H.G.Rammensee, J.Bachiuann position is L F E A M N N K N 4 and S.Stevanovic. The probability of being specified, 8 A M N K N C Q A 4 processed and presented is given in order to predict the length 6 F E A M N N K N C 3 of each T-cell epitopes. 2 L E N L F E A M N 2 peptide is 9 7 E A M N N K N C Q 2 amino Table ΧΓΧΒ, part 1: MHC Class I decamer acids, the analysis of 213P1F11 v.l (aa 1-242). end position P13P1F11 v.l: HLA-A*0201 decamers for each peptide is the start position plus eight 213P1F11V.6: HLA-B*2709 nonamers | Pos 1 2 3 4 5 6 7 8 9 score 1 K L E N L F E A M 10 Portion of 9 M N N K N C Q A L 10 SEQ ID 3 E N L F E A M N N 4 NO: 13; each start 4 N L F E A M N N K 3 position is 8 A M N N K N C Q A 2 specified, 6 F E A M N N K N C 1 the length of each peptide is 9 amino acids, the end position for each peptide is the start position plus eight E13P1F11 v.6: HLA-B*4402 nonamers Pos 1 2 3 4 5 6 7 8 9 score F E A M N N K N C 13 Portion of M N N K N C Q A L 13 SEQ ID L E N L F E A M 11 NO: 13; E A M N N K N C Q each start position is K L E N L F E A M specified, A M N N K N C Q A the length E N L F E A M N N of each N L F E A M N N K peptide is 9 L F E A M N N K N amino acids, the |end position| for each peptide is the start position plus eight 242 213P1F11 v.l: HLA-A*0201 decamers bl3PlFll v.l: HLA-A*0201 decamers Pos 1 2 3 4 5 6 7 8 9 0 score Pos 1 2 3 4 5 6 7 8 9 0 score 149 V M V I K D S P Q T 14 179 R H D Q K G S C F I 8 163 T D A L H V Y S T V 14 189 Q T L V D V F T K R 8 170 s T V E G Y I A Y R 14 191 L V D V F T K R K G 8 218 L V Q E G K A R K T 14 199 K G H I L E L L T E 8 222 G K A R K T N P E I 14 203 L Ξ L L T E V T R R 8 230 E I Q S T L R K R L 14 207 ' T E V T R R M A E A 8 6 S L E E E K Y D M S 13 43 F R .Q L R F E S T M 7 85 V L M A H G R E G F 13 76 E D P V S C A F V V 7 102 M V K L E N L F Ξ A 13 81 C A F V V L M A H G 7 134 G E Q R D P G E T V 13 110 E A L N N K N C Q A 7 167 H V Y S T V E G Y I 13 137 R D P G E T V G G D 7 175 Y I A Y R H D Q K G 13 141 E T V G G D E I V M 7 204 E L L T E V T R R M 13 147 E I V M V I K D S P 7 233 S T L R K R L Y L Q 13 148 I V M V I K D S P Q . 7 17 A R L A L I L C V T 12 160 P. T Y T D A L H V Y 7 99 D G E M V K L E N L 12 176 I A Y R H D Q K G S 7 104 K L E N L F E A L N 12 188 I Q T L V D V F T K 7 . 193 D V F T K R K G H I 12 214 A E A E L V Q E G K 7 E K Y D M S G A R L 11 25 V T K A R E G S E E 6 14 M S G A R L A L I L 11 42 M F R Q L R F E S T 6 38 A L E H M F R Q L R 11 72 I D S R E D P V S C 6 41 H M F R Q L R F Ξ S 11 87 M A H G R E G F L K 6 45 Q L R F E S T M K R 11 89 H G R E G F L K G E 6 51 T M K R D P T A E Q 11 97 G E D G E M V K L E 6 71 A I D S R E D P V S 11 101 E M V K L E N L F E 6 77 D P V S C A F V V L 11 114 N K N C Q A L R A K 6 78 P V S C A F V V L M 11 125 K V Y I I Q A C R G 6 159 I P T Y T D A L H V 11 139 P G E T V G G D E I 6 183 K G s C F I Q T L V 11 145 G D E I V M V I D 6 194 V F T K R K G H I L 11 152 I K D S P Q T I P T 6 63 E E L E K F Q Q A I 10 166 L H V Y S T V E G Y 6 79 V S C A F -V V L A 10 168 V Y S T V E G Y I A 6 122 A P K V Y I I Q A 10 171 T V E G Y I A Y R H 6 128 I I Q A C R G E Q R 10 186 C F I Q T L V D V F 6 140 G Ξ T V G G D E I V 10 206 L T E V T R R M A E 6 153 K D s P Q T I P T Y 10 215 E A E L V Q E G K A 6 158 T I P T Y T D A L H 10 7 L E E E K Y D M S G 5 190 T L V D V F T K R K 10 11 K Y D M s G A R L A 5 210 T R R M A ,E A E L V 10 24 C V T K A R E G S E 5 232 Q S T L R K R L Y L 10 29 R E G S E E D L D A 5 1 M S N P R s L E E E 9 32 S E E D L D A L E H 5 R S L E E E K Y D M 9 33 E E D L D A L E H M 5 D L D A L E H M F R 9 48 F E S T M K R D P T 5 73 D S R E D P V S C A 9 83 F V V L M A H G R E 5 84 V V L M A H G R E G 9 88 A H G R E G F L K G 5 93 G F L K G E D G E M 9 112 L N N K N C Q A L R 5 151 V I K D S P Q T I P 9 115 K N C Q A L R A K P 5 213 M A E A E L V Q E G 9 118 Q A L R A K P K V Y .5 217 E L V Q E G K A R K 9 121 R A K P K V Y I I Q 5 225 R K T N P E I Q S T 9 123 K P K V Y I I Q A C 5 S G A R L A L I L C 8 133 R G E Q R D P G E T 5 50 S T M K R D P T A E 8 136 Q R D P G E T V G G 5 64 E L E K F Q Q A I D 8 144 G G D E I V M V I K 5 80 S C A F V V L M A H 8 155 . S P Q T I P T Y T D 5 243 244 Q13P1F11 v.l: HLA-A*0202 decamers P13P1F11 v.l: HLA-A*0203 decamers Pos 1 2 3 4 5 6 7 8 9 0 score 215 E A E L V Q E G K A 2 223 K A R K T N P E I Q 2 17 A R L A L I L C V T 1 A L I L C V T K A R 1 28 A R E G S E E D L D 1 38 A L E H M F R Q L R 1 58 A E Q F Q E E L E K 1 71 A I D S R E D P V S 1 82 A F V V L M A H G R 1 88 A H G R E G F. L K G 1 1 11 A L N N K N C Q A L 1 1 19 A L R A K P K V Y I 1 122 A P K V Y I I Q A 1 131 A C R G E Q R D P G 1 165 A L H V Y s T V Ξ G 1 177 A Y R H D Q K G S C 1 216 A Ξ L V Q E G K A R 1 224 A R K T N P E I Q S 1 213P1F11 v. l : HLA-A*0203 decamers Pos 1 2 3 4 5 6 7 8 9 0 score 207 T E V T R R M A E A 18 Portion of 8 E E E K Y D M S G A 10 SEQ ID 11 K Y D M S G A R L A 10 NO: 3; each start 19 L A L I L C V T K A 10 position is 29 R E G S, E E D L D A 10 specified, 49 E S T M K R D P T A 10 the length 62 Q El E L E K F Q Q A 10 of each 73 D S R E D P V S c A 10 peptide is 79 V S C A F V V L M A 10 10 amino 102 M V K L E N L F E A 10 acids, the end posit E A L N N K ion 1 10 N C Q A 10 for each 1 13 N N K N C Q A L R A 10 peptide is 122 A K P K V Y I I Q A 10 the start 156 P Q T I P T Y T D A 10 position 168 V Y S T V E G Y I A 10 plus nine 205 L L T E V T R R M A 10 215 E A E L V Q E G K A 10 9 E E K Y D M S G A R 9 12 Y D M S G A R L A L 9 A L I L C V T K A R 9 . E G S E E D L D A L 9 50 S T M K R D P T Ά E 9 63· E E L E K F Q Q A I 9 74 S R E D P V S c A F 9 80 S C A F V V L M A H 9 103 V K L E N L F E A L 9 1 11 A L N N K N C Q A L 9 1 14 N K N C Q A L R A K 9 123 K P K V Y I I Q A C 9 157 Q T I P T Y T D A L 9 169 Y S T V E G Y I A Y 9 206 L T Ξ V T R R M A E 9 208 E V T R R M A E A E 9 245 E13P1F11 v.l: HLA-A1 decamers E13P1F11 v.l: HLA-A1 decamers Pos 1 2 3 4 5 6 7 8 9 0 score Pos 1 2 3 4 5 6 7 8 9 0 score 191 L V D V F T K R K G 12 49 E S T M K R D P T A 4 215 E A Ξ L V Q E G K A 12 65 L E K F Q Q A I D S 4 D L D A L E H M F R 11 77 D P V S c A F V V L 4 61 F Q E E L E K F Q Q 11 87 M A H G R E G F L K 4 64 E L E K F Q Q A I D 11 103 V K L E N L F E A L 4 90 G R E G F L K G E D 11 107 N L F E A L N N K N 4 108 L F E A L N N K N C 11 154 D S P Q T I P T Y T 4 139 P G E T V G G D E I 11 175 Y I A Y R H D Q K G 4 171 T V E G Y I A Y R H 11 13 D M S G A R L A L I 3 184 G S C F I Q T L V D 11 19 L A L I L C V T K A 3 189 Q T L V D V F T K R 11 37 D A L E H M F R Q L 3 202 I L E L L T E V T R 11 42 M F R Q L R F E S T 3 213 M A E A E L V Q E G 11 45 Q L R F E S T M K R 3 232 Q S T L R K R L Y L 11 55 D P T A E Q F Q E E 3 7 L E E E K Y D M s G 10 78 P V S C A F V V L M 3 8 E E E K Y D M S G A 10 85 V L M A H G R E G F 3 14 M S G A R L A L I L 10 115 K N C Q A L R A K P 3 33 E E D L D A L E H M 10 127 Y I I Q A C R G E Q 3 47 R F Ξ S T M K' R D P 10 131 A C R G E Q R D P G 3 99 D G Ξ M V K L E N L 10 143 V G G D E I V M V I 3 133 R G E Q R D P G E T 10 178 Y R H D Q K G S C F 3 144 G G D E I V M V I K 10 197 K R K G H I L E L L 3 157 Q T I P T Y T D A ,L 10 205 L L T E V T R R M A 3 179 R H D Q K G S C F I 10 2 S N P R S L E E E K 2 226 K T N P E I Q S T L 10 17 A R L A L I L C V T 2 233 S T L R K R L Y L Q 10 20 A L I L C V T K A R 2 12 Y D M S G A R L A L 9 48 F E s T M K R D P T 2 1 . M S< N P R S L E E E 8 59 E Q F Q E E L E K F 2 S G A R L A L I L C 8 80 S C A F V V L M A H 2 V T K A R E G S E E 8 84 V V L M A H G R E G 2 50 s T M K R D P T A E 8 100 G E M V K L E N L F 2 121 R A K P K V Y I I Q 8 111 A L N N K N C Q A L 2 170 S T V E G Y I A Y R 8 112 L N N K N C Q A L R 2 181 D Q K G S C F I Q T 8 117 C Q A L R A K P K V 2 198 R K G H I L E L L T 8 119 A L R A K P K V Y I 2 58 A E Q-F Q E E L E K 7 137 R D P G E T V G G D 2 101 E M V K L E N L F E 7 · 142 T V G G D E I V M V 2 209 V T R R M A E A E L 7 155 S P Q T I P T Y T D 2 211 R R M A E A E L V Q 7 158 T I P T Y T D A L H 2 16 G A R L A L I L C V 6 165 A L H V Y S T V E G 2 29 R E G S E E D L D A 6 168 V Y S T V E G Y I A 2 39 L E H M F R Q L R F 6 183 K G S C F I Q T L V 2 56 P T A E Q F Q E E L 6 185 S C F I Q T L V D V 2 98 E D G E M V K L E N 6 186 C F I Q T L V D V F 2 105 L E N L F E A L N N 6 192 V D V F T K R K G H 2 113 N N K N C Q A L R A 6 194 V F T K R K G H I L 2 159 I P T Y T D A L H V 6 210 T R R M A E A E L V 2 196 T K R K G H I L E L 6 216 A E L V Q E G K A R 2 199 K G H I L E L L T E 6 218 L V Q E G K A R K T 2 73 D S R E D P V S C A 5 227 T N P E I Q S T L R . 2 94 F L K G E D G E M V 5 229 P E I Q S T L R K R 2 122 A K P K V Y I I Q A 5 10 E K Y D M s G A R L 1 224 A R K T N P E I Q S 5 18 R L A L I L C V T K 1 R S L E E E K Y D M 4 22 I L C V T K A R E G 1 246 P13P1F11 v.l: HLA-A1 decamers 247 213PlFll v.l : HLA-A26 decamers Q13P1F11 v.l: HLA-A26 decamers Pos 1 2 3 4 5 6 7 Θ 9 0 score Pos 1 2 3 4 5 6 7 8 9 0 score 191 L V D V F T K R K G 12 82 A F V V L M A H G R 7 201 H I L E L L T E V T 12 89 ■ H G R E G F L K G E 7 206 L T E V T R R M A E 12 97 ' G E D G E M V K L E 7 A L I L C V T K A R 1 1 101 E M V K L E N L F E 7 . 39 L E H M F R Q L R F 1 1 108 L F E A L N N K N C 7 83 F V V L M A H G R E 1 1 110 E A L N N K N C Q A 7 84 V V L M A H G R E G 1 1 121 R A K P K V Y I I Q 7 106 E N L F E A L N N K 1 1 122 A K P K V Y I I Q A 7 125 K V Y I I Q A- C R G 1 1 123 K P K V Y I I Q A C 7 128 I I Q A C R G E Q R 1 1 135 E Q R D P G E T V G 7 148 I V M V I K D S P Q 1 1 185 S C F I Q T L V D V 7 167 H V Y S T V E G Y I 1 1 200 G H I L E L L T E V 7 187 F I Q T L V D V F T 1 1 215 E A E L V Q E G K A 7 195 F T K R K G H I L E 1 1 221 E G K A R K T N P E 7 205 L L T E V T R R M A 1 1 36 L D A L E H M F R Q 6 R S L E E E K Y D M 10 62 Q E E L E K F Q Q A 6 12 Y D M S G A R L A L 10 120 L R A K P K V Y I I 6 18 R L A L I L C V T K 10 143 V G G D E ■I V M V I 6 27 K A R E G S E E D L 10 163 T D A L H V Y S T V 6 40 E H M F R Q L R F E 10 164 D A L H V Y S T V E 6 45 Q L R F E s T M K R 10 1 73 E G Y I A Y R H D Q 6 63 E E L E K F Q Q A I 10 207 T E V T R R M A E A 6 100 G E M V K L E N L F 10 213 M A E A Έ L V Q E G 6 1 18 Q A L R A K P K V Y 10 225 R K T N P E I Q 3 T 6 165 A L H V Y s T V E G 10 15 S G A R L A L I L c 5 190 T L V D V F T K R K 10 19 L A L I L C V T K A 5 23 1 I Q S T L R K R L Y 10 79 V S C A F V V L M A 5 22 I L' C V T K A R E G 9 88 A H G R E G F L K G 5 38 A L E H M F R Q L R 9 1 14 N K N C Q A L R A K 5 43 F R Q L R F E S T M 9 136 Q R D P G E T V G G 5 46 L R F E S T M K R D 9 145 G D E I V M V I K D 5 67 K F Q Q A I D S R E 9 172 V E G Y I A Y R H D 5 86 L M A H G R E G F L 9 203 L E L L T E V T R R 5 92 E G F L K G E D G E 9 7 L E E E K Y D M S G 4 98 E D G E M V K L E N 9 16 G A R L A L I L C V 4 104 K L E N L F E A L N 9 17 A R L A L I L C V T 4 1 19 A L R A K P K V Y I 9 53 K R D P T A E Q F Q 4 202 I L E L L T E V T R 9 75 R E D P V S C A F V 4 2 12 R M A E A E L V Q E 9 132 C R G E Q R D P G E 4 9 E E K Y D M S G A R 8 156 P Q T I P T Y T D A 4 14 M S G A R L A L I L 8 188 I Q T L V D V F T K 4 60 Q F Q E E L E K F Q 8 199 K G H I L E L L T E 4 81 C A F V V L M A H G 8 214 A E A E L V Q E G K 4 137 R D P G E T V G G D 8 3 1 G S E E D L D A L E 3 138 D P G E T V G G D E 8 41 H M F R Q L R F E S 3 144 G G D E I V M V I K 8 51 T M K R D P T A E Q 3 154 D S P Q T I P T Y T 8 61 F Q E E L E K F Q Q 3 229 P E I Q s T L R K R 8 70 Q A I D S R E D P V 3 232 Q S T L R K R L Y L 8 90 G R E G F L K G E D 3 1 M S N P R s L E E E 7 95 L K G E D G E M V K 3 47 R F E S T M K R D P 7 1 15 K N C Q A L R A K P 3 49 E S T M K R D P T A 7 1 17 C Q A L R A K P K V 3 76 E D P V S C A F V V 7 126 V Y I I Q A C R G E 3 80 S C A F y V L M A H 7 129 I Q A C R G E Q R D 3 248 I213P1F11 v.l: HLA-A26 decamers 249 1213P1F11 v.l: HLA-A3 decamers B13P1F11 v.l: HLA-A3 decamers Pos 1 2 3 4 5 .6 7 8 9 0 score Pos 1 2 3 4 5 6 7 8 9 0 score 32 s E E D L D A L E H 12 54 R D P T A E Q F Q E 7 72 I D S R E D P V S C 12 73 D S R E D E V S c A 7 83 F V V L M A H G R E 12 74 S R E D P V s C A F 7 102 M V K L E N L F E A 12 76 E D P V S C A F V V 7 106 E N L F E A L N N K 12 91 R E G F L. K G E D G 7 187 F I Q T L V D V F T 12 105 L E N L F E A L N N 7 204 E L L T E V T R R M 12 112 L N N K N C Q A L R 7 E Y D M S G A R L 11 115 K N C Q A L R A K P 7 17 A R L A L I L C V T 11 124 P K V Y I I Q A C R 7 107 N L F E A L N N K N 11 137 R D P G E T V G G D 7 114 N N C Q A L R A K 11 164 D A L H V Y s T V E 7 151 V I K D S P Q T I P 11 169 Y S T V E G γ A Y 7 178 Y R H D Q K. G S c F 11 184 G S C F I Q T L V D 7 186 C F I Q T L V D V F 11 198 R K G H I L E L L T 7 193 D V F T K R K G H I 11 219 V Q E G K A R K T N 7- 199 K G H I L E L L T E 11 223 K A R K T N P E I Q 7 75 R E D P V S C A F V 10 224 A R K T N P E I Q S 7 82 A F V V L M A H G R 10 225 R K T N P E I Q s T 7 134 G E Q R D P G E T V 10 229 P E I Q s T L R K R 7 147 E I V M V I K D S P 10 232 Q S T L R K R L Y L 7 191 L V D V F T K R K G 10 11 K Y D M S G A R L A 6 3 N P R S L E E E Y 9 37 D A L E H M F R Q L 6 V T K A R E G S Ξ E 9 53 K R D P T A E Q F Q 6 88 A H G R E G F L K G 9 62 Q E E L E K F Q Q A 6 131 A C R G E Q R D P G 9 66 E K F Q Q A I D s R 6 135 E Q R D P G E T V G 9 67 K F Q Q A I D S R E 6 136 Q R D P G E T V G G 9 70 Q A I D S R E D P V 6 157 Q T I P T Y T D A L 9 79 V S C A F V V L M A 6 163 T D A L H V Y S T V 9 110 E A L N N K N C Q A 6 170 S T V E G Y I A Y R 9 122 A K P K V Y I I Q A 6 175 Y I A Y R H D Q K G 9 133 R G E Q R D P G E T 6 176 I A Y R H D Q K G S 9 141 E T V G G D E I V M 6 177 A Y R H D Q K G S C 9 143 V G G D E I V M V I 6 189 Q T L V D V F T K R • 9 166 L H V Y S T V E G Y 6 209 V T R R M A E A E L 9 173 E G Y I A Y R H D Q 6 230 E I Q S T L R K R L 9 181 D Q K G S c F I Q T 6 14 M s G A R L A L I L 8 182 Q K G S C F I Q T L 6 34 E D ' L D A L E H M F 8 185 S C F I Q T L V D V 6 43 F R Q L R F E S T M 8 196 T K R K G H I L E L 6 77 D P V S C A F V V L 8 197 K R K G H I L E L L 6 80 S C A F V V L M A H 8 210 T R R M A E A E L V 6 96 K G E D G E M V K L 8 227 T N P E I Q S T L R 6 113 N N K N C Q A L R A 8 233 s T L R K R L Y L Q 6 121 R A K P K V Y I I Q 8 12 Y D M S G A R L A L 5 159 I P T Y T D A L H V 8 13 D M S G A R L A L I 5 203 L E L L T E V T R R 8 15 S G A R L A L I L C 5 231 I Q S T L R K R L Y 8 28 A R E G S E E D L D 5 R S L E E E K Y D M 7 42 M F R Q L R F E S T 5 9 E E K Y D M S G A R 7 69 Q Q A I D S R E D P 5 16 G A R L A L I L C V 7 123 K P K V Y I I Q A C 5 27 K A R E G S E E D L 7 130 Q A C R G E Q R D P 5 29 R E G S E E D L D A 7 138 D P G E T V G G D E 5 39 L E H M F R Q L R F 7 155 S P Q T I P T Y T D 5 51 T M K R D P T A E Q 7 161 T Y T D A L H V Y S 5 250 251 213P1F11 v.l: HLA-B*0702 decamers 213P1F11 v.l: HLA-B*0702 decamers Pos 1 2 3 4 5 6 7 8 9 0 score Pos 1 2 3 4 5 6 7 8 9 0 score 152 I K D S P Q T I P T 10 88 A H G R E G F L K G 6 183 K G S C F I Q T L V 10 102 M V K L E N L F Ξ A 6 198 R K G H I L E L L T 10 134 G E Q R D P G E T V 6 42 M F R Q L R F E S T 9 139 P G E T V G G D E I 6 73 D S R E D P V S C A 9 140 G E T V G G D E I V 6 85 V L M A H G R E G F 9 167 H V Y S T V E G Y I 6 120 L R A K P K V Y I I 9 178 Y R H D Q K G S c F 6 122 A P K V Y I I Q A 9 193 D V F T K R K G H I 6 143 V G G D E I V M V I 9 211 R R M A E A E L V Q 6 162 Y T D A L H V Y s T 9 218 L V Q E G K A R K T 6 179 R H D Q K G S c F I 9 71 A I D S R E D P V S 5 181 D Q K G S C F I Q T 9 72 I D S R E D P V S C 5 187 F I Q T L V D V F T 9 101 E M V K L E N L F E 5 8 E E E K Y D M s G A 8 135 E Q R D P G E T V G 5 11 K Y D M S G A R L A 8 136 Q R D P G E T V G G 5 19 L A L I L C V T K A 8 153 K D S P Q T I P T Y 5 33 E E D L D A L E H M 8 165 A L H V Y S T V E G 5 39 L E H M F R Q L R F 8 18 R L A L I L C V T K 4 49 E S T M K. R D P T A 8 20 A L I L C V T K A R 4 52 M K R D P T A E Q F 8 28 A R E G S E E D L D 4 63 E E L E K F Q Q A I 8 50 S T M K R D P T A E 4 76 E D P V S C A F V V 8 ' 53 K R D -P T A E Q F Q 4 94 F L K G E D G E M V 8 58 A Ξ Q F Q E E L E K 4 1 13 N N K N C Q A L R A 8 97 G Ξ D G E M V K L E 4 117 C Q A L R A K P K V 8 98 E D G E M V K L E N 4 131 A c R G E Q R D P G 8 137 R D P G E T V G G D 4 168 V Y S T V E G Y I A 8 177 A Y R H D Q K G S C 4 185 S c F I Q T L V D V 8 184 G S C F I Q T L V D 4 186 C F I Q T L V D V F 8 212 R M A E A E L V Q E 4 201 H I L E L L T E V T 8 221 E G K A R K T N P E 4 204 E L L T E V T R R M 8 223 K A R K T N P E I Q 4 210 T R R M A E A E L V 8 35 D L D A L E H M F R 3 222 G K A R K T N P E I 8 38 A L E H M F R Q L R 3 R S L E E E K Y D M 7 40 E H M F R Q L R F E 3 34 E D L D A L E H M F 7 45 Q L R F E S T M K R 3 59 E Q F Q E E L E K F 7 64 E L E K F Q Q A I D 3 62 Q E E L E K F Q Q A 7 115 K N C Q A L R A K P 3 70 Q A I D S R E D P V 7 144 G G D E I V M V I K 3 74 s R E D P V S C A F 7 148 I V M V I K D S P Q 3 93 G F L K G E D G E M 7 199 K G H I L E L L T E 3 100 G E M V K L E N L F 7 202 I L E L L T E V T R 3 1 10 E A L N N K N C Q A 7 214 A E A E L V Q E G K 3 133 R G E Q R D. P G E T 7 216 A E L V Q E G K A R 3 149 V M V I K D S P Q T 7 219 V Q E G K A R K T N 3 1 50 M V I D S P Q T I 7 220 Q E G K A R K T N P 3 154 D S P Q T I P T Y T 7 231 I Q S T L R K R L Y 3 156 P Q T I P T Y T D A 7 4 P R S L E E E K Y D i 163 T D A L H V Y s T V 7 9 E E K Y D M S G A R 2 200 G H I L E L L T E V 7 32 S E E D L D A L E H 2 205 L L T E V T R R M A 7 44 R Q L R F E S T M K 2 207 T E V T R R M A E A 7 51 T M K R D P T A E Q 2 215 E A E L V Q E G K A 7 69 Q Q A I D S R E D P 2 225 R K T N P E I Q s T 7 80 S c A F V V L M A H 2 43 F R Q L R F E S T M 6 82 A F V V L M A H G R 2 252 253 213P1F11 v.l: HLA-B 402 decamers 213P1F11 v.l : HLA-B*4402 decamers Pos 1 2 3 4 5 6 7 8 9 0 score Pos 1 2 3 4 5 6 7 8 9 0 score 77 D P V S C A F V V L 13 50 s T M K R D P T A E 5 105 L Ξ N L F E A L N N 13 53. K R D P T A E Q F Q 5 150 M V I K D S P Q T I 13 71 A I D s R E D P V s 5 169 Y s T V E G Y I A Y 13 76 E D P V S C A F V V 5 203 L E L L T E V T R R 13 82 A F V V L M A H G R 5 214 A Ξ A E L V Q E G K 13 92 E G F L K G E D G E 5 232 Q S T L R K R L Y L 13 106 E N L F E A L N N K 5 3 N P R S L E E E K Y 12 107 N L F E A L N N K N 5 8 E Ξ E K Y D M S G A 12 110 E A L N N K N C Q A 5 13 D M S G A R L A L I 12 114 N K N C Q A L R A K 5 14 M S G A R L A L .1 L 12 131 A C R G E Q R D P G 5 A L I L C V T K A R 12 141 E T V G G D E I V M 5 65 L Ξ K F Q Q A I D S 12 142 T V G G D E I V M V 5 85 V L M A H G R E G F 12 165 A L H V Y S T V E G 5 109 F E A L N N K N C Q 12 185 S C F I Q T L V D V 5 119 A L R A K P K V Y I 12 200 G H I L E L L T E V 5 134 G E Q R D P G E T V 12 219 V Q E G K A R K T N 5 27 K A R E G s E E D L 11 233 S T L R K R L Y L Q 5 29 R E G S E E D L D A 11 1 M S N P R S L E E E 4 99 D G E M V K L E N L 11 4 P R S L E E E K Y D 4 143 V G G D E I V M V I 11 41 H M F R Q L R F E S 4 166 L H V Y S T V E G Y 11 46 L R F E S T M K R D 4 172 V E G Y I A Y R H D 11 70 Q A I D S R E D P V 4 178 Y R H D Q K G S C F 11 72 I D S R E D P V S C 4 193, D V F T K R K G H I 11 78 P V S C A F V V L M 4 194 V F T K R K. G H I L 11 80 s C A F V V L M A H 4 207 T E V T R R M A E A 11 113 N N K N C Q A L R A 4 209 V T R R M A E A Ξ L 11 116 N C Q A L R A K P K 4 7 L E E E K Y D M S G 10 121 R A K P K V Y I r Q 4 56 P T A E Q F Q E E L 10 126 V Y I I Q A C R G E 4 86 L M A H G R E G F L 10 127 Y I I Q A C R G E Q 4 91 R E G F L K G Έ D G 10 135 E Q R D P G E T V G 4 140 G Ξ T V G G D E I V 10 155 S P Q T I P T Y T D 4 220 Q E G K. A R K T N P 10 170 S T V E G Y I A Y R 4 120 L R A K P K V Y I I 9 173 E G Y I A Y R H D Q 4 122 A K E K V Y I I Q A 9 174 G Y I A Y R H D Q K 4 139 P G E T V G G D Ξ I 9 181 D Q K G S C F I Q T 4 179 R H D Q K G S C F I 9 190 T L V D V F T K R K 4 222 G K A R K T N P E I 9 199 K G H I L E L L T E 4 167 H V Y S T V E G Y I 8 215 E A E L V Q E G K A 4 66 E K F Q Q A I D S R 7 221 E G K A R K T N P E 4 88 A H G R E G F L K G 7 225 R T N P E I Q S T 4 17 A R L A L I L C V T 6 2 S N P R S L E E E K 3 123 K P K V Y I I Q A C 6 18 R L A L I L C V T K 3 136 Q R D P G E T V G G 6 19 L A L I L C V T K A 3 204 E L L T E V T R R M 6 49 E S T M K R D P T A 3 208 E V T R R M A E A E 6 54 R D P T A. E Q F Q E 3 224 A R K T N P E I Q S 6 89 H G R E G F L K G E 3 11 K Y D M S G A R L A 5 98 E D G E M V K L E N 3 S G A R L A L I L C 5 101 E M V K L E N L F E 3 16 G A R L A L I L C V 5 104 K L E N L F E A L N 3 28 A R E G S E E D L D 5 115 K N C Q A L R A K P 3 38 A L E H M F R Q L R 5 137 R D P G E T V G G D 3 40 E. H M F R Q L R F E 5 . 145 G D E I V M V I D 3 254 255 P13P1F11 v.2: HLA-A*0201 dccamers E13P1F11 v.2: HLA-A*0203 decamers 256 257 Q13P1F11 v.2: HLA-A3 decamers E13P1F11 v.2: HLA-B*0702 decamers 258 213P1F11 v.2: HLA-B*4402 decamers P13P1F11 v.3: HLA-A*0203 decamers TABLE XIXB, part 3: MHC Class 1 decamer anal sis of 213P1F11 v.3 (aa 1-146).

B13P1F11 v.3: HLA-A1 decamers Q13P1F11 v.3: HLA-A*0202 decamers [213P1F11 v.3: HLA-A26 decamers 259 260 513P1F11 v.4: HLA-A*0201 decamers 213P1F11 v.4: HLA-A*0202 decamers 261 213P1F11 v.4: HLA-A1 decamcrs 262 213P1F11 v.4: HLA-A3 decamers 263 213P1F11 v.4: HLA-B*0702 decamers 264 265 1 213P1F11 v.5: HLA-A3 decamers TABLE XIXB, part 6: MHC Class I decamer Pos 1 2 3 4 5 6 7 8 9 0 score analysis of 213P1F11 v.6 (aa 1 -242). 4 R L A L I L R V T K 35 Portion of 213P1F11 v.6: HLA-A*0201 decamers 6 A L I L R V T K A R 22 SEQ ID 8 I L R V T K A R E G 18 NO: 1 1 ; E G S E each start R V T K A R 17 position is 7 L I L R V T K A R E 16 spec i tied, 3 A R L A L I L R V T 11 the length 1 S G A R L A L I L R 9 of each 2 G A R L A L I L R V 7 peptide is L A L I L R V T K A 4 10 amino 9 L R V T K A R E G S 1 acids, the end position for each peptide is the start position plus nine 213P1F11 v.5: HLA-B*0702 decamers Pos 1 2 3 4 5 6 7 8 9 0 score 2 G A R L A L I L R V 10 Portion of 3 A R L A L I L R V T 10 SEQ ID L A L I L R V T K A 8 NO: 11 ; each start 4 R I A L I L R V T K 5 position is 6 A L I L R V T K A R 4 specified, 8 I L R V T K A R E G 3 the length R V T K A R E G S E 2 of each 1 S G A R L A L I L R 1 peptide is 7 L I L R V T K A R E 1 10 amino 9 L K V T K A R E G S 1 acids, the End position for each peptide is the start position plus nine 266 267 TABLE XKC: MHC Class Π Analysis of 213P1F11. Listed are scores which correlate with the ligation strengtli to a defined HLA type for a sequence of amino acids. The algorithms used are based on the book "MHC Ligands and Peptide Motifs" by H.G.Rammensee, J.Bachmann and S.Stevanovic. The probability of being processed and presented is given in order to predict T-cell epitopes.

Table X1XC, part 1 : MHC Class II 15-mer anal sis of 213P1F11 v. l a a 1 -242 . 268 213P1F11 v.l: HLA-DRB1*010115 -mers \ 269 213P1F11 v. l : HLA-DRB1 *0101 15 - niers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 47 R F E S T M K R D P T A E Q F 10 57 T A E Q F Q E E L E K F Q Q A 10 62 Q E E L E F Q Q A I D S R E 10 65 , L E K F Q Q A I D S R E D P V 10 71 A I D S R E D P V S C A F V V 10 94 F L K G E D G E M V K L E N L 10 1 15 K N C Q A L . R A K P K V Y I I 10 1 18 Q A L R A K P K V Y I I Q A C 10 128 I I Q A C R G E Q R D P G E T 10 138 D P G Ξ T V G G D E I V M V I 10 143 V G G D E I V M V I K D S P Q 10 172 V E G Y I A Y R H D Q K G S c 10 189 Q T L V D V F T K R K G H' I L 10 198 R K G H I L E L L T E V T R R 10 212 R M A E A E L V Q E G K A R K 10 215 E A E L V Q E G K A R K T N P 10 22 1 E G K A R K T N P E I Q S T L 10 3 N P R S L E E E K Y D M S G A 9 27 K A R E G S E E D L D A L E H 9 31 G S E E D L D A L E H M F R Q 9 D L D A L E H M F R Q L R F E 9 37 D A L E H M F R Q L R F E S T 9 41 H M F R Q L R F E S T M K R D 9 48 F E S T M K R D P T A E Q F Q 9 59 E Q F Q E Ξ L E K F Q Q A I D 9 60 Q F Q E E L E K F Q Q A I D S 9 75 R E D P V S C A F V V L M A H 9 85 V L M A H G R E G F L K G E D 9 105 L E N L F E A L N N K N C Q A 9 1 13 N N K N C Q A L R A K P K V Y 9 134 G E Q R D P G E T V G G D E I 9 139 P G E T V G G D E I V M V I K 9 161 T Y T D A L H V Y S T V E G Y 9 173 Ξ G Y I A Y R H D Q K G S C F 9 181 D Q K G S c F I Q T L V D V F 9 195 F T K R K G H I L E L L T E V 9 205 L L T E V T R R M A E A E L V 9 210 T R R M A E A E L V Q E G K A 9 222 G K A R K T N P E I Q S T L R 9 224 A R K T N P E I Q S T L R K R 9 , E K Y D M S G A R L A L I L C 8 13 D M S G A R L A L I L C V T K 8 S G A R L A L I L C V T K A R - 8 28 A R E G S E E D L D A L E H M 8 32 S E E D L D A L E H M F R Q L 8 42 M F R Q L R F E S T M R D P 8 56 P T A E Q F Q E E L E K F Q Q 8 61 F Q E E L E K F Q Q A I D S R 8 68 F Q Q A I D S R Ξ D P V S C A 8 77 D P V S C A F V V L M A H G R 8 98 E D G E M V K L E N L F E A L 8 101 E M. V K L Ξ N L F E A L N N K 8 108 L F E A L N N K N C Q A L R A 8 1 12 ' L N N K N C Q A L R A K P K V 8 125 K V Y I I Q A C R G E Q R D P 8 270 213P1F11 v.l: HLA-DRB1*0101 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 130 Q A C R G E Q R D P G E T V G 8 136 Q R D P G E T V G G D E I V M 8 141 E T V G G D E I V M V I K D S 8 156 P Q T I P T Y T D A L H V Y S 8 160 P T Y T D A L H V Y S T V E G 8 180 H D Q K G S C F I Q T L V D V 8 186 C F I Q T L V D V F T K R K G 8 220 Q E G K A R K T N P E I Q S T 8 7 L E E E K Y D M S G A R L A L 7 24 C V T K A R E G S E E D L D A 7 78 P V S C A F V V L M A H G R E 7 116 N C Q A L R A K P K V Y I I Q 7 122 A K P K V Y I I Q A C R G E Q 7 152 I K D s P Q T I P T Y T D A L 7 170 s T V E G Y I A Y R H D Q K G 7 219 V Q E G K A R K T N P E I Q S 7 46 . L R F E S T M K R D P T A E Q 6 64 Ξ L E K F Q Q A I D s R E D P 6 70 Q A I D S R E D P V s C A F V 6 79 V S C A F V V L M A H G R E G 6 96 K G E D G E M V K L E N L F E 6 13 1 A C R G E Q R D P G E T V G G 6 142 T V G G D E I V M V I K D S P 6 150 M V I K D S P Q T I P T Y T D 6 151 V I K D S P Q T I P T Y T D A 6 196 T K R G H I L E L L T E V T 6 87 M A H G R E G F L K G E D G E 5 190 T L V D V F T K R K G H I L E 5 90 G R E G F L K G E D G E M V K 4 ' 133 R G E Q R D P G Ξ T V G G D E 4 2 S N P R S L E E Ξ K Y D M S G 3 110 E A L N N K N C Q A L R A K P 3 164 D A L H V Y S T V E G Y I A Y 3 171 T V E G Y I A Y R H D Q K G S 3 201 H I L E L L T E V T R R M A E 3 38 A L E H M F R Q L R F E S T M 2 86 L M A H G R E G F L K G E D G 2 129 I Q A C R G E Q R D P G E T V 2 135 E Q R D P G E T V G G D E I V 2 167 H V Y S T V E G Y I A Y R H D 2 178 Y R H D Q K G S C F I Q T L V 2 187 F I Q T L V D V F T K R K G H . 2 193 D V F T K R K G H I L E L L T 2 223 K A R K T N P E I Q S T L R K 2 227 T N P E I Q S T L R K R L Y L 2 26 T K A R E G S E E D L D A L E 1 34 E D L D A L E H M F R Q L R F 1 44 R Q L R F E S T M K R D P T A 1 52 M K R D P T A E Q F Q E E L E 1 53 K R D P T A E Q F Q E E L E K 1 93 G F L K G E D G E M V K L E N 1 95 L K G E D G E M V K L E N L F 1 104 K L E N L. F E A L N N K N C Q 1 107 N L F Ξ A L N N K N C Q A L R 1 1 19 A L R A K P K V Y I I Q A C R 1 271 213P1F11 v.l: HLA-DRB1*010115 - mcrs 272 213P1F11 v.l: HLA-DRB1*0301 (DR17) 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5' score 43 F R Q L R F E S T M K R D P T 13 71 A I D S R E D P V S c A F V V 13 82 Ά F V V L M A H G R E G F L K 13 101 E M V K L E N L F E A L N N K 13 - 187 F I Q T L V D V F T K R K G H 13 210 T R R M A E A E L V Q E G K A 13 216 A E L V Q E G K A R K T N P E 13 4 P R S L E E E K Y D M S G A R 12 7 L E E E K Y D M S G A R L A L 12 11 K Y D M S G A R L A L I L C V 12 69 Q Q A I D S R E D P V S C A F 12 76 E D P V S C A F V V L M A H G 12 81 C A F V V L M A H G R E G F L 12 100 G E M V K L E N L F E A L N N 12 156 P Q T I P T Y T D A L H V Y S 12 163 T D A L H V Y S T V E G Y I A 12 173 E G Y I A Y R H D Q K G s C F 12 202 I L E L L T E V T R R M A E A 12 206 L T E V T R R M A E A E L V Q 12 A L I L C V T K A R E G S E E 11 31 G S E E D L D A L E H M F R Q 11 33 E E D L D A L E H M F R Q L R 11 D L p A L E H M F R Q L R F E 11 61 F Q E E L E K F Q Q A I D S R 11 75 R E D P V S C A F V V L M A H 11 91 R E G F L K G E D G E M V K L 11 92 Ξ G F L K G E D G E M V K L E 11 94 F L K G E D G E M V K L E N L 11 97 G E D G E M V K L E N L F E A 11 116 N C Q A L R A K P K V Y I I Q 11 132 C R G E Q R D P G E T V G G D 11 146 D E I V M V I K D S P Q T I P 11 147 E I V M V I K D S P Q T I P T 11 155 S P Q T. I P .T Y T D A L H V Y 11 167 H V Y S T V E G Y I A Y R H D 11 185 S C F I Q T L V D V F T K R K 11 188 I Q T L V D V F T K R K G H I 11 3 N P R S L E E E K Y D M S G A 10 8 E E E K Y D M S G A R L A L I 10 12 Y D M s G A R L A L I L C V T 10 28 A R E G S E E D L D A L E H M 10 57 T A E Q F Q E E L E K F Q Q A 10 123 K P K V Y I I Q A C R G E Q R 10 . 126 V Y Γ I Q A C R G E Q R D P G 10 169 Y S T V E G Y I A Y R H D Q K 10 180 H D Q K G S C F I Q T L V D V 10 194 y F T K R K G H I L E L L T E 10 195 F T K R K G H I L E L L T E V 10 40 E H M F R Q L R F E S T M K R 9 50 S T M R D P T A E Q F Q E E 9 72 I D S R E D P V S C A F V V L 9 80 s C A F V V L M A H G R E G F 9 108 L F E A L N N K N C Q A L R A 9 113 N N K N C Q A L R A K P K V Y 9 151 V I K D S P Q T I P T Y T D A 9 273 274 213P1F11 v.l: HLA-DRB1*0301 (DR17) 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 168 V Y S T V E G Y I A Y R H D Q 3 193 D V F T K R K G H I L E L L T 3 198 R K G H I L E L L T E V T R R 3 219 V Q E G K A R K T N P E I Q S 3 223 K A R K T N P E I Q S T L R K 3 S G A R L A L I L C V T K A R 2 23 L C V T K A R Έ G S E E D L D 2 47 R F E S T M K R D P T A E Q F 2 53 . K R D P T A E Q F Q E E L E K 2 79 V S C A F V V L M A H G R Έ G 2 85 V L M A H G R E G F L K G E D 2 87 M A H G R Ξ G F L K G E D G E 2 111 A L N N K N C Q A L R A K P K 2 119 A L R A K P K V Y I I Q A C R 2 138 D P G E T V G G D E I V- M V I 2 142 T V G G D E I V M V I K D S P .2 157 Q T I P T Y T D A L H V Y S T 2 162 Y T D A L M V Y S T V E G Y I 2 186 c F I Q T L V D V F T K R K G. 2 197 K R K G H I L E L L T E V T R 2 205 L L T E V T R R M A E A E L V 2 211 R R M A E A E L V Q E G K A R 2 .225 R K T N P Ξ I Q S T L R K R L 2 226 T N P E I Q S T L R K R L Y 2 227 T N P E I Q s T L R K R L Y L 2 R S L E E Ξ K Y D M S G A R L 1 13 D M S G A R L A L I L C V T K 1 24 C V T K A R E G S E E D L D A 1 E G S Ξ E D L D A L E H M F R 1 34 E D L D A L E H M F R Q L R F 1 44 R Q L R F E S T M K R D P T A 1 48 F E S T M K R D P T A E Q F Q 1 63 E E L E K F Q Q A I D S R E D 1 70 Q A I D S R E D P V S C A F V 1 77 D P V S C A F V V L M A H G R 1 78 P V s C A F V V L M A H G R E 1 89 H G R'' E G F L K G E D G E M V 1 95 L K G Ξ D G E M V K L E N L F 1 112 L N N K N C Q A L R A K P K V 1 118 Q A L R A K P K V Y I I Q A C 1 124 P K V Y I I Q A C R G E Q R D 1 130 Q A C R G E Q R D P G E T V G 1 136 Q R D P G E T V G G D E I V M 1 137 R D P G E T V G G D E I V M V 1 143 V G G D E I V M V I K D S P Q 1 144 G G D E I V M V I K D S P Q T 1 150 M V I K D s P Q T I P T Y T D 1 153 K D S P Q T I P T Y T D A L H 1 154 D S P Q T I P T Y T D A L H V 1 161 T Y T D A L H V Y s T V E G Y 1 166 L H V Y S T V E G Y I A Y R H 1 172 V E G Y i A Y R H D Q K G S C 1 177 A Y R H D Q K G S C F I Q T L 1 178 Y R H D Q K G S C F I' Q T L V 1 179 R H D Q K G S C F I Q T L V D 1 275 213P1F11 v.l: HLA-DRB1*0301 (DR17) 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 182 Q K G S C F I Q T L V D V F T 1 183 K G S C F I Q T L V D V F T K 1 221 E G K A R K T N P E I Q S T L 1 276 213P1F11 v.l: HLA-DRB 1*0401 (DR4Dw4) 15 - nicrs Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 84 V V L M A H G R E G F L K G E 14 123 K P K V Y I I Q A C R G E Q R 14 145 G D E I V M V I K D S P Q T I 14 149 V M V I K D S P Q T I P T Y T 14 163 T D A L H V Y S T V E G Y I A 14 165 A L H V Y S T V E G Y I A Y R 14 189 Q T L V D V F T K R K G H I L 14 200 G H I L E L L T E V T R R M A 14 210 T R R M A E A E L V Q E G K A 14 216 A E L V Q E G K A R K T N P E 14 N P R S L E E E K Y D M S G A 12 L E E E K Y D M S G A R L A L 12 E K Y D M S G A R L A L I L C 12 14 M S G A R L A L I L C V T K A 12 S G A R L A L I L C V T K A R 12 17 A R L A L I L C V T K A R E G 12 24 C V T A R E G S E E D L D A 12 27 K A R E G S E E D L D A L E H 12 28 A R E G S E E D L D A L E H M 12 31 G S E E D L D A L E H M F R Q 12 34 E D L D A L E H M F R Q L R F 12 37 D A L E H M F R Q L R F E S T 12 46 L R F E S T M K R D P T A E Q 12 52 M K R D P T A E Q F Q E E L E 12 54 R D P T A E Q F Q E E L E K F 12 59 E Q F Q E E L E K F Q. Q A I D 12 60 Q F Q E E L E K F Q Q A I D S 12 66 E K F Q Q A I D S R E D P V S 12 67 K F Q Q A I D S R E D P V S C 12 71 A I D S R E D P V S C A F V V 12 73 D S R E D P V S C A F V V L M 12 77 D P V S C A F V V L M A H G R 12 93 G F L K G E D G E M V K L E N 12 97 G E D G E M V K L E N L F E A 12 103 V K L E N L F E A L N N K N C 12 104 K L E N L F E A L N N K N C Q 12 108 L F E A L N N K N C Q A L R A 12 113 N N K N C Q A L R A K P K V Y 12 114 N K N C Q A L R A K P K V Y I 12 120 L R A K P V Y I I Q A C R G 12 122 A K P K V I I Q A C R G E Q 12 129 I Q A C R G E Q R D P G E T V 12 132 C R G E Q R D P G E T V G G D 12 137 R D P G E T V G G D E I V M V 12 141 E T V G G D E I V M V I K D S 12 142 T V G G D E I V M V I K D S P 12 150 M V I K D S P Q T I P T Y T D 12 153 K D S P Q T I P T Y T D A L H 12 155 S P Q T I P T Y T D A L H V Y 12 160 P T Y T D A L H V Y S T V E G 12 162 Y T D A L H V Y S T V E G Y I 12 174 G Y I A Y R H D Q K G S C F I 12 182 Q K G S C F I Q T L V D V F T 12 186 C F I Q T L V D V F T K R K G 12 196 T K R G H I L E L L T E V T 12 277 213P1F11 v.l: HLA-DRB1*0401 (DR4Dw4) 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 198 R K G H I L E L L T E V T R R 12 208 E V T R R M A E A E L V Q E G 12 209 V T R R M A E A E L V Q E G K 12 213 M A E A E L V Q E G K A R K T 12 225 R K T N P E I Q, S T L R K R L 12 226 K T N P E I Q S T L R K R L Y 12 192 V D V F T K R K G H I L E L L 11 45 Q L R F E s T M K R D P T A E 10 124 P K V γ I I Q A C R G E Q R D 10 184 G S C F I Q T L V D V F T K R 10 69 Q Q A I D s R E D P V S C A F 9 109 F E A L N N K N C Q A L R A K 9 117 C Q A L R A K P K V Y I I Q A 9 191 L V D V F T K R K G H I L E L 9 A L I L C V T K A R E G S E E 8 126 V Y I I Q A C R G E Q R D P G 8 .6 P. Q T I P T Y T D A L H V Y S 8 169 Y S T V E G Y I A Y R H D Q K 8 21 L I L c V T K A R E G S E E D 7 61 F Q E E L E K F Q Q A I D S R 7 177 A Y R H D Q K G S C F I Q. T L 7 217 E L V Q E G K A R K T N P E I 7 220 Q E G K A R K T N P E I Q S T 7 1 M S N P R S L E E E K Y D M S 6 2 S N P R ■S L Ξ E E K Y D M S G 6 6 S L E Ξ E K Y D M S G A R L A 6 8 E E E K Y D M S G A R L' A L I 6 13 D M S G A R L A L I L C V T K 6 V T K A R E G S E E D L D A L 6 29 R E G S E E D L D A L E H M F 6 Ξ G S E E D L D A L E H M F R 6 32 S E E D L D A L E H M F R Q L 6 D L D A L E H M F R Q L R F E 6 51 T M K R D P T A E Q F Q E E L 6 53 K R D P T A E Q F Q E E L E K 6 55 0 P T A E Q F Q E E L E K F Q 6 56 P T A E Q F Q E E L E K F Q Q 6 57 T A E Q F Q E E L E K F Q Q A 6 63 E E L E K F Q Q A I D S R E D 6 68 F Q Q A I D s R Ξ D P V S C A 6 70 Q A I D S R E D P V S c A F V 6 72 I D s R E D P V S C A F V V L 6 74 s R E D P V s C A F V V L M A 6 75. R E D P V S c A F V V L M A H 6 78 P V S C A F V V L M A H G R E 6 79 V S C A F V V L M A H G R E G 6 86 L M A H G R E G F L K G E D G 6 88 A H G R E G F L K G E D G E M 6 89 H G R Ξ G F L K G E D G E M V 6 94 F L K G E D G E M V K L E N L 6 95 L K G Ξ D G E M V K L E N L F 6 96 K G E D G E M V K L E N L F E 6 101 E M V K L E N L F E A L N N K 6 107 N L F E A L N N K N C Q A L R 6 110 E A L N N K N C Q A L R A K P 6 278 213P1F11 v.l: HLA-DRB1*0401 (DR4Dw4) 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 1 1 1 Ά L N N K N C Q A L R A K P K 6 112 L N N K N C Q A L R A K P K V 6 1 16 N C Q A L R A K P K V Y I I Q 6 1 18 Q A L R A K P K V Y I I Q A c 6 128 I I Q A C R G E Q R D P G E T 6 134 G E Q R D. P G E T V G G D E I 6 135 E Q R D P G E T V G G D E I V 6 138 0 P G E T V G G D E I V M V I 6 139 P G E T V G G D E I V M V I K 6 143 V G G D E I V M V I K D S P Q 6 144 G G D Ξ I V V I K D S P Q T 6 152 I K D S P Q T I P T Y T D A L 6 154 D S P Q T I P T Y T D A L H V 6 157 Q T I P T Y T D A L H V Y S T 6 158 T I P T Y T D A L H V Y S T V 6 161 T Y T D A L H V Y S T V E G Y 6 167 H V Y S T V E G Y I A Y R H D 6 170 S T V E G Y I A Y R H D Q K G 6 178 Y R H D Q K G S c F I Q T. L V 6 179 R H D Q K G S C F I Q T L V D 6 180 H D Q G S C F I Q T L V D V 6 183 K G S C F I Q T L V D V F T K 6 187 F I Q T L V D V F T K R K G H 6 190 T L V D V F T K R K G H I L E 6 194 V F T K R K G H I L E L L T E 6 195 F T K R K G H I L E L L T E V 6 197 K R K G H I L E L L T E V T R 6 204 ' E L L T E V T R R M A E A E L 6 207 T E V T R R M A E A E L V Q E 6 21 1 R R M A E A E L V Q E G K A R 6 218 L V Q E G K A R K T N P E I Q 6 221 E G K A R K T N P E I Q S T L . 6 222 G K A R K T N P E I Q S T L R 6 223 K A R K T N P E I Q S T L R K 6 173 E G Y I A Y R H D Q K G S C F 3 206 L T E V T R R M A E A E L V Q 3 12 Y D M s G A R L A L I L C V T 1 41 H M F R Q L R F E S T M K R D 1 47 R F E S T M K R D P T A E Q F 1 85 V L M A H G R E G F L K G E D 1 1 15 K N C Q A L R A K P K V Y I I 1 1 19 A L R A K P K V Y I I Q A C R 1 13 1 A C R G E Q R D P G E T V G G 1 193 p V F T K' R K G H I L E L L T 1 205 L L T E V T R R M A E A E L V 1 219 V Q E G K A R K T N P E I Q S 1 R S L E E E K Y D M S G A R L . -5 23 L C V T K A R E G S E E D L D -5 38 A L E H M F R Q L R F E S T M -5 48 F E S T M K R D P T A E Q F Q -5 90 G R E G F L K G E D G E M V K -5 98 E D G E M V L E N L F E A L -5 127 Y I I Q A C R G E Q R D P G E -5 279 213P1F11 v.l: HLA-DRB1*1101 15 - mers Portion of SEQ ID NO: 3; each start position is specified, the length of each peptide is 15 amino acids, the end position for each peptide is the start position plus fourteen 280 213P1F11 v.l: HLA-DRB1*1101 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 200 G H I L E L L T E V T R R M A 12 124 P K V Y I I Q A C R G E Q R D 11 184 G s c F I Q T L V D V F T K R 11 202 I L E L L T E V T R R M A E A 11 58 A E Q F Q E E L Ξ K F Q Q A I 10 91 R E G F L K G E D G E M V K L 10 115 K N C Q A L R A K P K V Y I I 9 142 T V G G D Ξ I V M V I K D S P 9 193 D V F T K R K G H I L E L L T 9 204 E L L T E V T R R M A E A E L 9 3 N P R S L E E E K Y D M S G A 8 8 E E E K Y D M S G A R L A L I 8 E K y D M S G A R L A L I L C 8 18 R L A L I L C V T K A R E G S 8 22 I L c V T A R E G S E E D L 8 47 R F E s T M K R D P T A E Q F 8 77 D P V s C A F V V L M A H G R 8 78 P V s c A F V V L M A H G R E 8 88 A H G R E G F L K G E D G E M 8 105 L E N L F E A L N N K N C Q A 8 107 N L F E A L N N K N C Q A L R 8 111 A L N N K N C Q A L R A K P K 8 154 D S P Q T I P T Y T D A L H V . 8 165 A L H V Y s T V E G Y I A Y R 8 169 Y S T V E G Y I A Y R H D Q K 8 171 T V E G Y I A Y R H D Q K G S 8 186 C F I Q T L V D V F T K R K G 8 190 T L V D V F T K R K G H I L E 8 206 L T E V T R R M A E A E L V Q 8 212 R M A E A E L V Q E G K A R K 8 216 A E L V Q E G K A R K T N P E 8 217 E L V Q E G K A R K T N P E I 8 1 M S N P R S L E E E K Y D M S 7 11 K Y D M S G A R L A L I L C V 7 13 D M S G A R L A L I L C V T K 7 S G A R L A L I L C V T K A R 7 43 F R Q L R F E s T M K R D P T 7 62 Q E E L E K F Q Q A I D S R E 7 66 E K F Q Q A I D S R E D P V S 7 76 E D P V S C A F V V L M A H G 7 86 L M A H G R E G F L K G E D G 7 128 I I Q A C R G E Q R D P G E T 7 133 R G E Q R D P G E T V G G D E 7 143 V G G D E I V M V I K D S P Q 7 156 P Q T I P T Y T D A L H V Y S 7 162 Y T D A L H V Y S T V E G Y I 7 170 S T V Ξ G Y I A Y R H D Q K G 7 182 Q K G S C F I Q T L V D V F T 7 196 T K R K G H I L E L L T E V T 7 225 R K T N P E I Q S T L R K R L 7 R s L E E E K Y D M S G A R L 6 6 S L E E E K Y D M S G A R' L A 6 27 K A R E G S E E D L D A L E H 6 E G S E E D L D A L E H M F R 6 49 E S T M K R D P T A E Q F Q E 6 281 213P1F11 v. l: HLA-DRB1*1101 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 60 Q F Q E E L E K F Q Q A I D S ■ 6 63 E E L Ξ K F Q Q A I D S R E D 6 71 A I D S R E D P V s C A F V V 6 73 D S R E D P V S C A F V V L M 6 84 V V L M A H G R E G F L K G E 6 92 E G F L K G E D G E M V L E 6 97 6 E D G E M V K L E N L F E A 6 108 L F E A L N N K N C Q A L R A 6 109 F E A L N N K N C Q A L R A K 6 114 N K N C Q A L R A K P K V Y I 6 122 A K P K V Y I I Q A C R G E Q 6 126 V Y I I Q A C R G E Q R D P G 6 134 G E Q R D P G E T V G G D E I 6 137 R D P G E T V G G D E I V M V 6 140 G E T V G G D E I V M V I K D 6 147 E I V M V I K D s P Q T I P T 6 148 I V M V I K D S P Q T I P T Y 6 153 K D S P Q T I P T Y T D A L H 6 160 P T Y T D A L H V Y s T V E G 6 174 G Y I A Y R H D Q K G S C F I 6 197 K R K G H I L E L L T E V T R . 6 205 L L T E V T R R M A E A E L V 6 207 T E V T R R M A E A E L V Q E 6 210 T -R R M A E A E L V Q E G K A 6 21 1 R R M A E A E L V Q E G K A R 6 222 G K A R K T N P E I Q S T L R 6 138 D P G Ξ T V G G D E I V M V I 4 187 F I Q T L V D V F T K R K G H 4 44 R Q L R F E S T M K R D P T A ; 3 1 19 A L R A K P K V Y I I Q A C R 3 167 H V Y S T V E G Y I A Y R H D 3 34 E D L D A L E H F R Q L R F 2 37 D A L E H M F R Q L R F E S T 2 53 K R D P T A E Q F Q E E L E K 2 57 T A E Q F Q E E L E K F Q Q A 2 72 I D S R E D P V S C A F V V L 2 74 s R E D P V S C A F V V L M A 2 98 E D G E M V K L Ξ N L F E A L 2 127 Y I I Q A C R G E Q. R D' P G E 2 136 Q R D P G E T V G G D E I V M 2 157 Q T I P T Y T D A L H V Y S T 2 161 T Y T D A L H V Y S T V E G Y 2 ■ 201 H I L E L L T E V T R R M A E 2 223 K A R K T N P E I Q S T L R K 2 226 K T N P E I Q S T L R K R L Y 2 227 T N P Ξ i Q S T L R K R L Y L 2 12 Y D M S G A R L A L I L C V T 1 26 T K A R E G S E E D L D A L E 1 29 R E G S E E D L D A L E H M F 1 D L D A L E H M F R Q L R F E 1 38 A L E H M F R Q L R F E S T M 1 55 D P T A E Q F Q E E L E K F Q 1 64 E L E K F Q Q A I D S R E D P 1 75 R E D P V S C A F V V L M A H 1 85 V L M A H G R E G F L K G E D 1 282 213P1FH Λ : HLA-DRB1*110115 - mers 283 213P1F11 v.2: HLA-DRB OIOI 15 - mers 284 213P1F11 v.2: HLA-DRB1*0301 (DR17) 15 - niers 285 213P1F11 v.2: HLA-DRB1*0401 (DR4Dw4) 15 - meis Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 22 L Y L P S E A P P N P P L W N 6 P S E A P P N P P L W N S Q D 6 26 S E A P P .N P P L W N S Q D T 6 27 E A P P N P P L W N S Q D T S 6 34 L W N S Q D T S P T D M I R K 6 37 S Q D T S P T D M I R K A H A 6 3 D T S P T D M I R K A H A L S 6 47 R K A H A L S R P W W M C S R 6 52 L S R P W W C S R R G K D I 6 41 S P T D M I R K A H A L S R P 1 56 W W M C S R R G K D I S W N F 1 48 K A H A L S R P W W M C S R R -5 286 TABLE XIXC, part 3: MHC Class II 15-mer analysis of 213P1F11 v.3 (aa 1-146). 213P1F11 v.3: HLA-DRB1*0101 15 - mers 213P1F11 v.3: HLA-DRB1*0301 (DR17) 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 6 V Y I I Q A C R G A T L P S P 19 Portion of K V Y I I Q A C R G A T L P S 18 SEQ ID 12 c R G A T L P S P F P Y L s L 11 NO: 7; each start 3 K P K V Y I I Q A C R G A T L 10 position is Q A C R G A T L P S P F P Y L 9 specified, 2 A K P K V Y I I Q A C R G A T 8 the length 1 R A K P K V Y I I Q A C R G A 3 of each 7 Y I I Q A C R G A T L P S P F 2 peptide is 4 P K V Y I I Q A C R G A T L P 1 15 amino 8 I I Q A C R G A T L P S P F P 1 acids, the 9 ■ I end Q A C R G A T L P S P F P Y 1 position for 11 A C R G A T L P S P F P Y L S 1 each peptide is the start position plus fourteen 287 213P1F11 v.3: HLA-DRB1*0401 (DR4Dw4) 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score K V Y I I Q A C R G A T L P S 26 Portion of 4 P K V Y I I Q A C R G A T L P 16 SEQD K P K V Y I I Q A C R G A T L 14 NO: 7; 3 each start 6 V Y I I Q A C R G A T L P S P 14 position is 2 A K P K V Y I I Q A C R G A T 12 specified, 8 I I Q A C R G A T L P S P F P 12 the length 11 A C R G A T L P S P F P Y L S 12 of each 9 I Q A C R G A T L P S P F P Y 6 peptide is 7 Y I I Q A C R G A T L P S P F -5 15 amino acids, the end position for each peptide is the stall position plus fourteen 213P1F11 v.3: HLA-DRB1*110115 - mers 288 TABLE XDiC, part 4: MHC Class II 15-mer analysis of 213P1F1 1 v,4 (aa 1-321). 213P1F11 v.4: HLA-DRB1 *0101 15 - mers Portion of SEQ ID NO: 9; each start position is specified, the length of each peptide is 15 amino acids, the end position for each peptide is the start position plus fourteen 289 213P1F11 v.4: HLA-DRB1*01()1 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 .1 2 3 4 5 score . 43 R G S S V H Q K L V N D P R E 7 47 V H Q K L V N D P R E T Q E V 7 60 E V F G G G V G D I V G R D L 7 69 I V G R D L S I S F R N S E T 7 76 I S F R N S E T S A S E E E K 7 86 s E E E K Y D M S G A R L A L 7 11 L S V Q P E K R T G L R D E N 6 38 Q G R A F R G S S V H Q K L V 6 78 F R N S E T S A S E E E K Y D 6 9 K S L S V Q P E K R T G L R D 3 13 V Q P E K R T G L R D E N G E 3 16 E K R T G L R D Ξ N G E C G Q 3 19 T G L R D E N G E C G Q T F R 3 54 D P R E T Q E V F G G G V G D 3 14 Q P E K R T G L R D E N G E C 2 22 R D E N G E C G Q T F R L K E 2 27 E C G Q T F R L K E E Q G R A 2 52 V N D P R E T Q E V F G G G V 2 61 V F G G G V G D I V G R D L S 2 80 N S E T S A S E E E K Y D M S 2 81 S E T S A S E E E K Y D M S G 2 6 E Y D K S L S V Q P E K R T G 1 17 K R T G L R D E N G E' C G Q T 1 G L R D E N G E C G Q T F R L 1 N G E C G Q T F R L K E . E Q G 1 26 G E C G Q T F R L K E E Q G R 1 62 F G G G V G D I V G R D L S I 1 79 ■ R N S E T S A S E E E K Y D M 1 213P1F11 v.4: HLA-DRB1 *0301 (DR17) 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 67 G D I V G R D L S I s F R N S 29 Portion of | 48 H Q K L V N D P R E T Q E V F 28 SEQ BD S L s V Q P E K R T G L R D E 26 NO: 9; T F R L K E E each start 3 1 Q G R A F R G S 21 position is j 17 K R T G L R D E N G E C G Q T 19 specified, 29 G Q T F R L K E Ξ Q G R A F R 18 the length 63 G G G y G D I V G R D L S I S 18 of each 8 D K S L S V Q P E K R T G L R 17 peptide is 71 G R D L S I S F R N S E T S A 17 15 amino 49 Q K L V N D P R E T Q E V F G 16 acids, the N S E T S A s E E E K Y D M S end 80 16 position for 51 L V N D P R E T Q E V F G G G 14 each 2 G K C Q E Y D K s L S V Q P E 12 peptide is 44 G S S V H Q K L V N D P R E T 12 the start 86 S E E E K Y D M s G A R L A L 12 position 18 R T G L R D E N G E C G Q T F 1 1 plus 40 R A F R G S S V H Q K L V N D 1 1 fourteen 58 T Q E V F G G G V G D I V G R 11 62 F G G G V G D I V G R D L S I 11 66 V G D I V G R D L S I S F R N 1 1 75 S I S F R N S E T S A S E E E 1 1 39 G R A F R G S S V H Q K L V N 10 73 D L S I S F R N S E T S A S E 10 21 L R D E N G E C G Q T F R L K 9 290 291 213P1F11 v.4: HLA-DRB1*0301 (DR17) 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 69 I V G R D L S I S F R N S E T 1 84 S A S E E E K Y D M S G A R L 1 213P1F11 v.4: HLA-DRB 1*0401 (DR4Dw4) 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score S L S V Q P E K R T G L R D E 26 Portion of 48 H Q K L V N D P R E T Q E V F 26 SEQID 75 S I S F R N S E T S A S E E E 22 NO: 9; 63 G G G V G D I V G R D L S I S eacli start position is 67 G D I V G R D L S I S F R N S 20 specified, 2 G K C Q E Y D K S L S V Q P E 18 the length Q T F R L K E E Q G R A F R G 18 of each K E E Q G R A F R G S S V H Q 18 peptide is 41 A F R G S S V H Q K L V N D P 18 15 amino 64 G G V G D I V G R D L S I S F 18 acids, the 72 R D L S I S F R N S E T S A S 18 end position for 4 C Q E Y D K S L S V Q P E K R 16 each 39 G R A F R G S S V H Q K L V N 16 peptide is 59 Q E V F G G G V G D I V G R D 16 the start 73 D L S I S F R N S E T S A S E 15 position 8 D K S L S V Q P E K R T G L R 14 pl s 18 R T G L R D E N G E C G Q T F 14 fourteen 31 T F R L K E E Q G R A F R G S 14 58 T Q E V F G G G V G D I V G R 14 71 G R D L S I S F R N S E T S A 14 6 E Y D K S L S V Q P E K R T G 12 14 Q P E K R T G L R D E N G E C 12 17 K R T G L R D E N G E C G Q T 12 23 D E N G E C G Q T F R L K E E 12 36 E E Q G R. A F R G S S V H Q K 12 38 Q G R A F R G S S V H Q K L V 12 40 R A F R G S S V H Q K L V N D 12 45 S S V H Q K L V N D P R E T Q 12 51 L V N D P R E T Q E V F G G G 12 55 P R E T Q E V F G G G V G D I 12 69 I V G R D L S I S F R N S E T 12 70 V G R D L S I S F R N S E T S 12 76 I S F R N S E T S A S E E E K 12 79 R N S E T S A S E E E K Y D M ' 12 82 E T S A S E E E K Y D M S G A 12 83 T S A S E E E K Y D M S G A R 12 86 S E E E K Y D M S G A R L A L 12 66 V G D I V G R D L S I S F R N 9 49 Q K L V N D P R E T Q E V F G 8 50 K L V N D P R E T Q E V F G G 7 1 G K C Q E Y D K S L S V Q P 6 Q E Y D K S L S V Q P E K R T 6 7 Y D K S L S V Q P E K R T G L 6 .P E K T G L R D E N G E C G 6 19 T G L R D E N G E C G Q T F R 6 G L R D E N G E C G Q T F R L 6 21 L R D E N G E C G Q T F R L K 6 •22 R D E N G E C G Q T F R L K E 6 24 E N G E C G Q T F R L K E E Q 6 N G E C G Q T F R L K E E Q G 6 292 213P1F11 v.4: HLA-DRB1*0401 (DR4Dw4) 15 - mers 293 213P1F11 v.4: HLA-DRB1M10115 - mei s Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score N G E C G Q T F R L K E E Q G 9 14 Q P E K R T G L R D E N G E C 8 Q T F R L K E E Q G R A F R G 8 32 F R L K E E Q G R A F R G S S 8 33 R L K E E Q G R A F R G S S V 8 45 S S V H Q K L V N D P R E T Q 8 46 S V H Q K L V N D P R E T Q E 8 56 R E T Q E V F G G G V G D I V 8 82 E T S A S E E E K Y D M S G A 8 7 Y D K S L S V Q P E K R T G L 7 31 T F R L K E E Q G R A F R G S 7 34 L K E E. Q G R A F R G S S V H 7 41 A F R G S S V H Q K L V N D P 7 58 T Q E V F G G G V G D I V G R 7 74 L S I S' F R N S E T S A S E E 7 Q E Y D K S L S V Q P E K R T 6 49 Q K L V N D P R E T Q E V F G 6 52 V N D P R E T Q E V F G G G V 6 53 N D P R E T Q E V F G G G V G 6 55 P R E T Q E V F G G G V G D I 6 67 G D I V G R D L S I S F N S . 6 68 D I V G R D L S I S F R N S E 6 83 T S A S E E E K Y D M S G A R 6 84 S A S E E E K Y D M S G A R L 6 85 A S E E E K Y D M S G A R L A 6 6 E Y D K S L S V Q P E K R T G 2 12 S V Q P E K R T G L. R D E N G 2 23 D E N G E C G Q T F R L K E E 2 24 E N G E C G Q T F R L K E E Q 2 47 V H Q K L V N D P R E T Q E V 2 62 TABLE XIXC, part 5: MHC Class II 15-mer analysis of 213P1F11 v.5 (aa 1-242). 213P1F11 v.5: HLA-DRB1*010115 -mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 8 A R L A L I L R V T K A R E G 21 Portion of 11 A L I L R V . T K A R E G S E E 19 SEQ ID 2 K Y D M S G A R L A L I L R V 16 NO: 11; Y D M S G A R L A L I L R V T 16 each start 3 position is 9 R L A L I L R V T K A R E G s 16 specified, 7 G A R L A L I L R V T k A R E 15 the length L A L I L R V T K A R E G S E 14 of each M S G A R L A L I L R V T K A 11 peptide is 12 L I L R V T K A R E G S E E D 11 15 amino 13 I L R V T K A R E G S E E D L 11 acids, the 14 L R V T K A R E G S E E D L D 10 end position for 6 S G A R L A L I L R V T k A R 9 each 1 E K Y D M S G A R L A L I L R 8 peptide is 4 D M S G A R L A L I L R V T K 8 the start R V T K A R E G S E E D L D A \ position pLus fourteen 294 213P1F11 v.5: HLA-DRB1*0301 (DR17) 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 7 G A R L A L I L R V T K A R E 19 Portion of Ii A L I L R V T K A R E G S E 19 SEQID 1 E K Y D M S G A R L A L I L R 17 NO: 11; I L R V T K A R E G S E E D L 17 each start 13 position is 9 L A L I L R V T K A R E G S 13 specified, 2 K Y D S G A R L A L I L R V 12 the length 11 A L I L R V T K A R E G S E E 12 of each 3 Y D M S G A R L A L I L R V T 10 peptide is 12 L I L R V T K A R E G S E E D 7 15 amino M S G A R L A L I L R V T K A 4 acids, the end 8 A R L A L I. L R V T K A R E G 3 position for 6 S G A R L A L I L R V T K A R 2 each 14 L R V T K A R E G S E E D L D 2 peptide is 4 D M S G A R L A L I L R V T K 1 the start R V T K A R E G S E E D L D A 1 position plus fourteen 213P1F11 v.5: HLA-DRB1*0401 (DR4Dw4) 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 7 G A R L A L I L R V T K A R E 26 Portion of 2 K Y D M S G A R L A L I L R V 20 SEQID L A L I L R V T K A R E G S E 14 NO: 11; each start 13 I L R V T K A R Ξ G S E E D L 14 position is 1 E K Y D M S G A R L A L I L R 12 specified, S G A R L A L I L R V T K A 12 the length 6 S G A R L A L I L R V T K A R 12 of each 8 A R L A L I L R V T K A R E' G 12 peptide is R V T A R E G S E E D L D A . 12 15 amino 9 R L A L I L R V T K A R E G S 9 acids, the end 11 A L I L R V T K A R E G S E E 8 position for 12 L I L R V T K A R E G S E E D 7 each 4 D M S G A R L A L I L R V T K 6 peptide is 3 Y D M S G A R L A L I L R V T 1 the start 14 L R V T K A R E G S E E D L D -5 position plus fourteen 295 213P1F11 v.5: HLA-DRB1 * 1101 15 - mers TABLE XIXC, part 6: MHC Class II 15-mer analysis of 213P1F11 v.6 (aa 1-242). 213P1F11 v.6: HLA-DRB1 *0101 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score 2 D G E M V K L E N L F E A M N 20 Portion of 3 G E M V K L E N L F E A M N N 18 SEQ ID 9 E N L F E A M N N K N C Q A L 17 NO: 13; each start 12 F E A M N N K N C Q A L R A K 17 position is 14 A M N N K N C Q A L R A K P K 17 specified, M V K L E N L F Ξ A M N N K N 16 the length 6 V K L E N L F E A M N N K N C 11 of each 8 L E N L F E A M N N K N C Q A 9 peptide is 1 E D G E M V K -L E N L F E A M 8 15 amino 4 E M V K L E N ■ L F E A M N N K 8 acids, the 11 L F E A M N N K N C end Q A L R A 8 position for M N N K N C Q A L R A K P K V 8 each 1'3 E A M N N K N C Q A L R A K P 3 peptide is 7 K L E N L F E A M N N K N C Q 1 the start N L F E A M N N K N C Q A L R 1 position plus fourteen 296 213P1F11 v.6: HLA-DRB1*0301 (DR17) IS - mers Pos ■ 1 2 3 4 5 6 7 β 9 0 1 2 3 4 5 score 12 F E A M N N K N C Q A L R A K 20 Portion of 2 D G E M V K L E N L F E A M N 19 SEQ E) 8 L E N L F E A M N N K N C Q A 19 NO: 13; each start 1 Ξ D G E M V K L E N L F E A M 17 position is M V K L E N L F E A M N N K N 17 ' specified, 9 E N L F E A M N N K N C Q A L 15 the length 3 G E M V K L E N L F E A M N N 12 of each • 10 N L F E A M N N K N C Q A L R 8 peptide is 13 Ξ A M N N K N C Q A L R A K P 8 15 amino 1 1 L F E A M N N K N C Q A L R A 7 acids, the N L F E A M N N K end 4 E M V K L E 5 position for 7 K L E N L F E A M N N K N C Q 4 each 6 V K L E N L F E A M N N K N C 3 peptide is 14 A M N N K N C Q A L R A K P K 2 the start position plus fourteen 213P1F11 v.6: HLA-DRB1*0401 (DR4Dw4) 15 - mers Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 5 score 8 L E N L F E A M N N K N C Q A 26 Portion of 2 D G E M V K L E N L F E A M N 20 SEQ ID 3 G E M V K L E N L F E A N N 20 NO: 13; each start M V K L E N L F E A M N N K N 20 position is 9 E N L F E A M N N K N C Q A L 16 specified, 6 V K L E N L F E A M N . N K N C 12 the length 7 K L E N L F E A M N N K N C Q 12 of each 11 L F E A M N N K N C Q A L R A 12 peptide is 12 F E A M N N K N C Q A L R A K 9 15 amino 4 E M V K L E N L F E A M N N K 6 acids, the end N L F E A M N N K N C Q A L R 6 position for 13 Ξ A M N N K N C Q A L R A K P 6 each 14 A M N N K N C Q A L R A K P K 6 peptide is M N N K N C Q A L R A K P K V 6 the start 1 E D G E M V K L E N L F E A M -5 position plus fourteen 297 213P1F11 v.6: HLA-DRB1*1101 15 - mers Pos 1 2 3 4 5 6 7 ' 8 9 0 1 2 3 4 5 score 9 E N L F E A M N N K N C Q A L 16 Portion of 2 D G E M V K L E N L F E A M N 12 SEQ ID 3 . G E M V K L E N L F E. A M N N 12 NO: 13; each start M V K L E N L F Ξ A M N N K N 12 position is 8 L E N L F E A M N N K N C Q A 8 specified, N L F E A M N N K N c Q A L R •8 the length 14 A M N N K N C Q A L R A K P K 8 of each 1 1 L F E A M N N K N C Q A L R A 6 peptide is 12 F E A M N N K N C Q A L R A K 6 1 5 amino 1 E D G E M V K L E N L F E A M 2 acids, the R A K P end 13 E A M N N N C Q A L 1 position for each . peptide is the start position plus fourteen 298 Table XX: Frequently Occurring Motifs avrg. % Name Description Potential Function identity Nucleic acid-binding protein Zinc finger, C2H2 functions as transcription factor, zf-C2H2 34% type nuclear location probable cytochrome b Cytochrome b(N- membrane bound oxidase, generate N 68% terminal)/b6/petB superoxide domains are one hundred amino Immunoglobulin acids long and include a conserved is 19% domain int adomain disulfide bond. tandem repeats of about 40 residues, each containing a Trp-Asp motif.

WD domain, G-beta Function in signal transduction and WD40 18% repeat protein interaction may function in targeting signaling PDZ 23% PDZ domain molecules to sub-membranous sites LRR 28% Leucine Rich Repeat short sequence motifs involved in protein-protein interactions conserved catalytic core common to both serine/threonine and tyrosine protein kinases containing an ATP pkinase 23% Protein kinase domain binding site and a catalytic site pleckstrin homology involved jn intracellular signaling or as PH 16% PH domain constituents of the cytoskeleton -40 amino-acid long found in the extracellular domain of membrane- bound proteins or in secreted EOF 34% EGF-like domain proteins Reverse transcriptase (RNA-dependent rvt 49% DNA polymerase) Cytoplasmic protein, associates integral membrane proteins to the ank 25% Ank repeat cytoskeleton NADH- Ubiquinone/plastoquin membrane associated. Involved in one (complex I), proton translocation across the oxidored q l 32% various chains membrane 299 Table XX, continued: Frequently Occurring Motifs avrg. % Name Description Potential Function identity calcium-binding domain, consists of al2 residue loop flanked on both sides by a 12 residue alpha-helical efhand 24% EF hand domain Retroviral aspartyl Aspartyl or acid proteases, centered rvp 79% protease on a catalytic aspartyl residue extracellular structural proteins involved in formation of connective tissue. The sequence consists of the Collagen triple helix G-X-Y and the polypeptide chains Collagen 42% repeat (20 copies) forms a triple helix.

Located in the extracellular ligand- binding region of receptors and is about 200 amino acid residues long Fibronectin type III with two pairs of cysteines involved fa3 20% domain in disulfide bonds seven hydrophobic transmembrane regions, with the N-terminus located 7 transmembrane extracellularly while the C-terminus receptor (rhodopsin is cytoplasmic. Signal through G 7tm 1 19% family) proteins 300 Table XXI: Motifs and Post-translational Modifications of 213P1F11 A) Post-translational Modifications of 213P1F11- Tyrosine sulfation site. - 19 rsleeekYdinsgarl Protein kinase C phosphorylation site 51-53 Tni 196 - 198 TkR 210-212 TrR 234 - 236 T1R Casein kinase II phosphorylation site 6- 9 SleE 32 - 35 SeeD 74 -77 SreD 161 - 164 TytD 170 - 173 StvE 190 - 193 TlvD 227 - 230 TnpE Tyrosine kinase phosphorylation site - 12 RsleEek.Y N-nryristoylation site 16-21 GArlAL Bipartite nuclear targeting sequence 211 -227 RRniaeaelvqegkarkt B) Post-translational Modifications of 213P1F11-v.2 Tyrosine sulfation site . - 19 rsleeekYdinsgarl Protein kinase C phosphorylation site. 51-53 TmK 220-222 SrR Casein kinase II phosphorylation site . 6- 9 SleE 32 - 35 SeeD 74 -77 SreD 161 - 164 TytD 170 - 173 StvE 201 -204 SptD Tyrosine kinase phosphorylation site - 12 RsleEek.Y N-myristoylation site 16-21 GArlAL 301 TABLE XXII: Protein Properties of 213P1F1 1 TABLE XXII, continued: Protein Properties of 213P1F11 Table XXIIIA. Exon compositions of'213PlFll v.l Table XXIVA. Nucleotide sequence of transcript variant 213P1F1 1 v.2 (SEQ. I'D. NO. 55). ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc- acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcotct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaaatgagca atccgoggtc 420 tttggaagag gagaaatatg atatgtcagg tgcccgcctg gccctaatac tgtgtgtcac 480 caaagcccgg gaaggttccg aagaagacct ggatgctctg gaacacatgt ttcggcagct 540 gagattcgaa agcaccatga aaagagaccc cactgccgag caattccagg aagagctgga 600 aaaattccag caggccatcg attccoggga agatcccgtc agttgtgcct tegtggtact 660 catggctcac gggagggaag gcttcctcaa gggagaagat ggggagatgg tcaagctgga 720 gaatctcttc gaggccctga acaacaagaa ctgcoaggcc ctgcgagcta agcccaaggt 780 gtacatcata caggcctgtc gaggagaa'ca aagggacccc ggtgaaacag taggtggaga 840 tgagattgtg atggtcatca aagacagccc acaaaccatc ccaacataca cagatgcctt 900 gcacgtttat tccacggtag agggacccac gcccttccag gatcccctct acctaccctc 960 tgaagctccc ccgaacccac ctctctggaa ttcccaggat acatcgccta ccgacatgat 1020 cagaaaggct catgctttat ccagaccctg gtggatgtgt tcacgaagag gaaaggacat 1080 atcttggaac ttctgacaga ggtgacccgg cggatggcag aagcagagct ggttcaagaa 1140 ggaaaagcaa ggaaaacgaa ccctgaaatc caaagcaccc tccggaaacg gctgtatctg 1200 cagtagaagt agaaagacca ggaggagctt tccttccagc attctttctg tctcacagaa 1260 atttagaggc agctcttacc tctccccaag atcttctgtt cccaaggcca aatggcaccc 1320 agtttctttt ccatcacacc cttcatgcag gtcctcctgt ccttattaga gcaagccagc 1380 caaaacttag cacaaggcat ggtggcaaca ttaacatcac ctccctcagg ctggactttc 1440 tatctttatt aatgcaaccg aagagaccta agagtgcatt cacttatccc actttctgtt 1500 cctgtggtct tctttctccc atgaagcaga aactggataa agctcaagat tttccataga 1560 caaaccaaag cccactcatc ccctcctacc ccaatccaac ctctgctggc tcctgcatct 1620 cacttggagg tcaaacctcc tcctgaggcc aatgcattcc caacttccag ttctttcctt 1680 taccctggag agttagtaag gtaagaacca ttctttctct ccaaaaccac tcctccttgg 1740 ctggcaagtt ggtgtcctaa ctccgttctc ttcctagctc atggcctctc tagataataa 1800 agttgtctcc tcctttctgg atctcttcct cctaacaccc ctcccctgaa accctggact 1860 ctgccctctc tccaagaaaa tccatctatt caactattct tgc'attcaat tactctaaat 1920 gagagcgtgt tggagctatg gcaaattccc tgttgtcacc ttgctatttt gcagacaaca 1980 taatatttaa cctctcataa ccagagaggt taaataattt gtcaaatgca atacagtaag 2040 acagaggcaa ggacaaggtt tgacttccag cccagcctct tttccacaac ctgctaaatc 2100 ctgatccatc tgaaaacttt tctaattagt gaagatgact aataaaaatt ttccctatct 2160 ccaaggtagg agctttctgg aagtttctag aaattttcaa taaccaccag ccaaggttac 2220 ctccaggtaa ccttgcagca ccaggctgga agtcagatcg gcttcactat cttccaactc 2280 tacagcctgt atctctccat ccccagcttt gacctttcct gctcaagtaa cctacgggca 2340 catccagcgt cactaaaaac tcagggcttt tcttcccggt tactcctcca agcgttccct · 2400 ggtatcctca acctcagatc ccaggttcag atttctgcag tcaatctatg acccctctct 2460 tcttgcatcc ttcatatgcc accagacacc atgcccagtc cagcctgatt ttgaaacaac 2520 tttcatgccg gtcttctctt ccctgacatg ttactgtcca ggctcaagtc ctcagcttct 2580 catatctgca tctttgcaac caacttcctc ccttgcctct ctgcttttcc atcccacttt . 2640 tcatgtgtcc tccataccat ctataacagt gatctccctg gaacactcaa gaagacacaa 2700 cataccatat tatttaaaga ccagggtact ggacagtggc tcacacctgt attcccgact 2760 ttgagagtct gaagcgggag gatcacttga ggccaggagt taagagacca. gcctgggcaa 2820 cacagcaaga ccctgtctct aaaaaaaaaa attaattaac tgggtatggt ggcacatgcc 2880 tgtagtccca gctactcagg aggctgaggt gggaggatga cttgagccca ggagtttgag 2940 gctgcaagga gctatgatca tgccagtgca tcccagctct aggtgagaca' gtgagatccg 3000 gtctccaaaa taaatcaatc aatcaaataa agaccaaagt caaaccgcac atcaggatct 3060 ctcacaccct tccaattttg ccatctacca gcacttagct aaacccatct cccatctctt 3120 ccaccatgaa ttcactcttt caaaaaggct aatgtcttct tactcaccct tgcctctaag 3180 cctttgctat caccatttcc cccaagctgg agggccctcc ctctcccttt acccctcttc 3240 cactacctcc cacccctact ttttccagaa agc'catttcc tctctttttt ctgattgatc 3300 cttccctctc acccaggatt agatgctgga aatgaccact tctggagggc agggaacaag 3360 cccttaatct gcataatgag tgttcaataa acagttgtca aactttgaaa 3410 Table XXIVB. Nucleotide sequence of transcript variant 213P1F11 v.3 (SEQ. ID. NO. 56). ctqactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gaoagaaaac tcaccgacaa taaagggcca ggtgattgtg -180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattoca caqtgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaaatgagca atccgcggtc 420 tttggaagag gagaaatatg atatgtcagg tgcccgcctg gccctaatac tgtgtgtcac 480 caaagcccgg gaaggttccg aagaagacct ggatgctctg gaacacatgt ttcggcagct 540 gagattcgaa agcaccatga aaagagacoc cactgccgag caattccagg aagagctgga 600 aaaattccag caggccatcg attcccggga agatcccgtc agttgtgcct tcgtggtact 660 catggctcac gggagggaag gcttcctcaa gggagaagat ggggagatgg tcaagctgga 720 gaatctcttc gaggccctga acaacaagaa ctgccaggcc ctgcgagcta agcccaaggt 780 gtacatcata caggcctgtc gaggagccac cctgcccagc ccctttcott acctttctct 840 ctgactttgc ctcctcctct tcttgttgtt tcagaacaaa gggaccccgg tgaaacagta 900 ggtggagatg agattgtgat. ggtcatcaaa gacagcccac aaaccatcoc aacatacaca 960 gatgccttgc acgtttattc cacggtagag ggatacatcg cctaccgaca tgatcagaaa 1020 ggctcatgct ttatccagac cctggtggat gtgttcacga agaggaaagg acatatcttg 1080 gaacttctga cagaggtgac ccggcggatg gcagaagcag agctggttca agaaggaaaa 1140 gcaaggaaaa cgaaccctga aatccaaagc accctccgga aacggctgta totgcagtag 1200 aagtagaaag accaggagga gctttccttc cagcattctt tctgtctcac agaaatttag 1260 aggcagctct tacctctccc caagatctto tgttcccaag gccaaatggc aoccagtttc 1320 ttttccatca caoccttcat gcaggtcctc ctgtcottat tagagcaagc cagccaaaac 1380 ttagcacaag gcatggtggc aacattaaca tcacctccct caggctggac tttctatctt 1440 tattaatgca accgaagaga cctaagagtg cattcactta tcccacttt.c tgttcctgtg 1500 gtcttctttc tcccatgaag cagaaactgg ataaagctca agattttcca tagacaaacc 1560 aaagcccact catcccctcc taccccaatc caacctctgc tggctcctgc atctcacttg 1620 gaggtcaaac ctcctcctga ggccaatgca ttcccaactt ccagttcttt cctttaccct 1680 ggagagttag taaggtaaga accattcttt ctctccaaaa ccactcctcc ttggctggca 1740 agttggtgtc ctaactccgt tctcttccta gctcatggcc tctctagata ataaagttgt 1800 ctcctccttt ctggatctct tcctcctaac acccctcccc tgaaaccctg gactctgccc 1860 tctctccaag aaaatccatc tattcaacta ttcttgcatt caattactct aaatgagagc 1920 gtgttggagc tatggcaaat tccctgttgt caccttgcta ttttgcagac aacataatat 1980 ttaacctctc ataaccagag aggttaaata atttgtcaaa tgcaatacag taagacagag 2040 gcaaggacaa ggtttgactt ccagcccagc ctcttttcca caacctgcta aatcctgatc 2100 catctgaaaa cttttctaat tagtgaagat gactaataaa aattttccct atctccaagg 2160 taggagcttt ctggaagttt ctagaaattt tcaataacca ccagccaagg ttacctccag 2220 gtaaccttgc agcaccaggc tggaagtcag atcggcttca ctatcttcca actctacagc 2280 ctgtatctct ccatccccag ctttgacctt tcctgctcaa gtaacctacg ggcacatcca 2340 gcgtcactaa aaactcaggg cttttcttcc cggttactcc tccaagcgtt ccctggtatc 2400 ctcaacctca gatcccaggt tcagatttct gcagtcaatc tatgacccct ctcttcttgc 2460 atccttcata tgccaccaga caccatgccc agtccagcct gattttgaaa caactttcat 2520 gccggtcttc tcttccctga catgttactg tccaggctca agtcctcagc ttctcatatc 2580 tgcatctttg caaccaactt cctcccttgc ctctctgctt ttccatccca cttttcatgt 2640 gtcctecata ccatctataa cagtgatctc cctggaacac tcaagaagac acaacatacc 2700 atattattta aagaccaggg tactggacag tggctcacac ctgtattccc gactttgaga 2760 gtctgaagcg ggaggatcac ttgaggccag gagttaagag accagcctgg gcaacacagc 2820 aagaccctgt ctctaaaaaa aaaaattaat taactgggta tggtggcaca tgcctgtagt 2880 cccagctact caggaggctg aggtgggagg atgacttgag cccaggagtt tgaggctgca 2940 aggagctatg atcatgccag tgcatcccag ctctaggtga gacagtgaga tccggtctcc 3000 aaaataaatc aatcaatcaa ataaagacca aagtcaaacc gcacatcagg atctctcaca 3060 cccttccaat tttgccatct accagcactt agctaaaccc atctcccatc tcttccacca 3120 tgaattcact ctttcaaaaa ggctaatgtc ttcttactca cccttgcctc taagcctttg 3180 ctatcaccat ttcccccaag ctggagggcc ctccctctcc ctttacccct cttccactac 3240 ctcccacccc tactttttcc agaaagccat ttcctctctt ttttctgatt gatccttccc 3300 tctcacccag gattagatgc tggaaatgac cacttctgga gggcagggaa caagccctta 3360 atctgcataa tgagtgttca ataaacagtt gtcaaacttt gaaa 3404 Table XXIVC. Nucleotide sequence of transcript variant 213P1F11 v.4 (SEQ. ID. NO. 57). atggggaaat gccaagagta tgacaaaagt ctgtctgtgc agccagagaa gagaacagga 60 ctcagagatg agaatggaga atgtggacag acattcagac tcaaggaaga gcaagggagg 120 gctttcaggg gaagttcagt ccaccagaag ctggtgaatg acccacggga gacacaggaa . 180 gtttttgggg g'cggagtggg ggacattgtg ggacgggatc tcagtattag cttcagaaac 240 tctgagacct ctgcaagtga ggaggagaaa tatgatatgt caggtgcccg cctggcccta 300 atactgtgtg tcaccaaagc ccgggaaggt tccgaagaag acctggatgc tctggaacac 360 atgtttcggc agctgagatt cgaaagcacc atgaaaagag accccactgc cgagcaattc 420 caggaagagc tggaaaaatt ccagcaggcc atcgattccc gggaagatcc cgtcagttgt 480 gccttcgtgg tactcatggc tcacgggagg gaaggcttcc tcaagggaga agatggggag 540 atggtcaagc tggagaatct cttcgaggcc ctgaacaaca agaactgcca ggccctgcga 600 gctaagccca aggtgtacat catacaggcc tgtcgaggag aacaaaggga ccccggtgaa 660 acagtaggtg gagatgagat tgtgatggtc atcaaagaca gcccacaaac catcccaaca .720 tacacagatg ccttgcacgt ttattccacg gtagagggat acatcgccta ccgacatgat 780 cagaaaggct catgctttat ccagaccctg gtggatgtgt tcacgaagag gaaaggacat 840 atcttggaac ttctgacaga ggtgacccgg cggatggcag aagcagagct ggttcaagaa 900 ggaaaagcaa ggaaaacgaa ccctgaaatc caaagcaccc tccggaaacg gctgtatctg 960 cagtag 966 Table XXV A. Nucleotide sequence alignment of 213P1F11 v.l (SEQ. I'D. NO.58) and 213P1F11 v.2 (SEQ. ID. NO.59). 213P1F11V.1 CTGACTCATTTAGACTCTCTGCCTAGGCCACCTTTGCCAGAGGGAGTCCCCTCAGCCTTG 60 213 PI Fl 1 v .2 C'TGACTCATTTAGACTCTCTGCCTAGGCCACCTTTGCCAGAGGGAGTCCCCTCAGCCTTG 60 ************************************************************ 213P1F11V.1 CGATCACTCATCCCATTGGCGTTGGCTCCATTTCCACACCACAGCTGTGTGCCAAGGGTG 120 213P1F11V.2 CGATCACTCATCCCATTGGCGTTGGCTCCATTTCCACACCACAGCTGTGTGCCAAGGGTG 120 ************************************************************ 213P1F11V.1 TGTCATGAGGTTTCTTGAGTGACAGAAAACTCACCGACAATAAAGGGCCAGGTGATTGTC 180 213P1 11V.2 TGTCATGAGGTTTCTTGAGTGACAGAAAACTCACCGACAATAAAGGGCCAGGTGATTGTG 1B0 ************************************************************ 213P1F11V.1 CCACCCGATTCATAGACCAGGCTTCTCAGGAGAAACCCCGGGAGATTCCACACTGTCAGC 240 213P1F11V.2 CCACCCGATTCATAGACCAGGCTTCTCAGGAGAAACCCCGGGAGATTCCACACTGTCAGC 240 ************************************************************ 213P1F11V.1 CCCTTC'TCCAAGATCAGTACGTGGGCCTGACTCCTCCTCGGTGCCCAGCTCAGTATTGGC 300 213P1F1 lv.2 CCCTTCTCCAAGATCAGTACGTGGGCCTGACTCCTCCTCGGTGCCCAGCTCAGTATTGGC 300 ************************************************************ 213P1F1 lv.1 . AACTAGGAGAGTAGTGAGATTGAACTTGGCCTTGAGGAACAGCTGCCTCTAGAGTTGGAT 360 213P1F11 .2 . AACTAGGAGAGTAGTGAGATTGAACTTGGCCTTGAGGAACAGCTGCCTCTAGAGTTGGAT 360 213P1F11V.1 CAGACAAGGGTGCTGAGAGCCGGGACTCACAACCAAAGGAGAAATGAGCAATCCGCGGTC 420 213P1F1 lv.2 CAGACAAGGGTGCTGAGAGCCGGGACTCACAACCAAAGGAGAAATGAGCAATCCGCGGTC 420 ************************************************************ 213P1F11V.1 TTTGGAAGAGGAGAAATATGATATGTCAGGTGCCCGCCTGGCCCTAATACTGTGTGTCAC 480 213P1F11V.2 TTTGGAAGAGGAGAAATATGATATGTCAGGTGCCCGCCTGGCCCTAATACTGTGTGTCAC 480 ************************************************************ 213P1F11V.1 CAAAGCCCGGGAAGGTTCCGAAGAAGACCTGGATGCTCTGGAACACATGTTTCGGCAGCT 540 213 P 1 Fl 1 v .2 CAAAGCCCGGGAAGGTT CCGAAGAAGACCT GGATGCT CTGGAACACATGTTT CGGCAGCT 5 0 ************************************************************ 213P1F11V.1 GAGATTCGAAAGCACCATGAAAAGAGACCCCACTGCCGAGCAATTCCAGGAAGAGCTGGA 600 213P1 Fl 1 .2 GAGATTCGAAAGCACCATGAAAAGAGACCCCACTGCCGAGCAATTCCAGGAAGAGCTGGA 600 ************************************************************ 213P1F11V.1 AAAATTCCAGCAGGCCATCGATTCCCGGGAAGATCCCGTCAGTTGTGCCTTCGTGGTACT 660 213P1F11V .2 AMATTCCAGCAGGCCATCGATTCCCGGGAAGATCCCGTCAGTTGTGCCTTCGTGGTACT 660 ************************************************.************ 213P1F11V.1 CATGGCTCACGGGAGGGAAGGCTTCCTCAAGGGAGAAGATGGGGAGATGGTCAAGCTGGA 720 213P1 l lv .2 CATGGCTCACGGGAGGGAAGGCTTCCTCAAGGGAGAAGATGGGGAGATGGTCAAGCTGGA 720 213P1F11V.1 GAATCTCTTCGAGGCCCTGAACAACAAGAACTGCCAGGCCCTGCGAGCTAAGCCCAAGGT 780 213 PI l 1 v .2 GAATCTCTTCGAGGCCCTGAACAACAAGAACTGCCAGGCCCTGCGAGCTAAGCCCAAGGT 780 ************************************************************ 213P1F11V.1 GTACATCATACAGGCCTGTCGAGGAGAACAAAGGGACCCCGGTGAAACAGTAGGTGGAGA 8 0 213P1F11V.2 GTACATCATACAGGCCTGTCGAGGAGAACAAAGGGACCCCGGTGAAACAGTAGGTGGAGA 840■ 213P1 Fl 1 v .1 TGAGATTGTGATGGTCATCAAAGACAGCCCACAAACCATCCCAACATACACAGATGCCTT 900 213 PI Fl 1 v .2 TGAGATTGTGATGGTCATCAAAGACAGCCCACAAACCATCCCAACATACACAGATGCCTT 900 ************************************************************ 213P1F11V.1 GCACGTTTATTCCACGGTAGAGGGA 925 213P1F11V.2 GCACGTTTATTCCACGGTAGAGGGACCCACGCCCTTCCAGGATCCCCTCTACCTACCCTC 960 ************************* / . : TACATCGCCTACCGACATGAT 946 TGAAGCTCCCCCGAACCCACCTCTCTGGAATTCCCAGGATACATCGCCTACCGACATGAT 1020 ********************* CAGAAAGGCTCATGCTTTATCCAGACCCTGGTGGATGTGTTCACGAAGAGGAAAGGACAT 100.6 CAGAAAGGCTCATGCTTTATCCAGACCCTGGTGGATGTGTTCACGAAGAGGAAAGGACAT 1080 ************************************************************ ATCTTGGAACTTCTGACAGAGGTGACCCGGCGGATGGCAGAAGCAGAGCTGGTTCAAGAA 1066 ATCTTGGAACTTCTGACAGAGGTGACCCGGCGGATGGCAGAAGCAGAGCTGGTTCAAGAA 1140 ************************************************************ GGAAAAGCAAGGAAAACGAACCCTGAAATCCAAAGCACCCTCCGGAAACGGCTGTATCTG 1126 GGAAAAGCAAGGAAAACGAACCCTGAAATCCAAAGCACCCTCCGGAAACGGCTGTATCTG 1200 ************************************************************ CAGTAGAAGTAGAAAGACCAGGAGGAGCTTTCCTTCCAGCATTCTTTCTGTCTCACAGAA 1186 CAGTAGAAGTAGAAAGACCAGGAGGAGCTTTCCTTCCAGCATTCTTTCTGTCTCACAGAA 1260 ************************************************************ ATTTAGAGGCAGCTCTTACCTCTCCCCAAGATCTTCTGTTCCCAAGGCCAAATGGCACCC 1246 ATTTAGAGGCAGCTCTTACCTCTCCCCAAGATCTTCTGTTCCCAAGGCCAAATGGCACCC 1320 ************************************************************ AGTTTCTTTTCCATCACACCCTTCATGCAGGTCCTCCTGTCCTTATTAGAGCAAGCCAGC 1306 AGTTTCTTTTCCATCACACCCTTCATGCAGGTCCTCCTGTCCTTATTAGAGCAAGCCAGC 1380 ************************************************************ CAAAACTTAGCACAAGGCATGGTGGCAACATTAACATCACCTCCCTCAGGCTGGACTTTC 1366 CAAAACTTAGCACAAGGCATGGTGGCAACATTAACATCACCTCCCTCAGGCTGGACTTTC 1 40 ************************************************************ TATCTTTATTAATGCAACCGAAGAGACCTAAGAGTGCATTCACTTATCCCACTTTCTGTT 1426 TATCTTTATTAATGCAACCGAAGAGACCTAAGAGTGCATTCACTTATCCCACTTTCTGTT 1500 ************************************************************ CCTGTGGTCTTCTTTCTCCCATGAAGCAGAAACTGGATAAAGCTCAAGATTTTCCATAGA 1486 CCTGTGGTCTTCTTTCTCCCATGAAGCAGAAACTGGATAAAGCTCAAGATTTTCCATAGA 1560 ************************************************************ CAAACCAAAGCCCACTCATCCCCTCCTACCCCAATCCAACCTCTGCTGGCTCCTGCATCT 1546 CAAACCAAAGCCCACTCATCCCCTCCTACCCCAATCCAACCTCTGCTGGCTCCTGCATCT 1620 ************************************************************ CACTTGGAGGTCAAACCTCCTCCTGAGGCCAATGCATTCCCAACTTCCAGTTCTTTCCTT 1606 CACTTGGAGGTCAAACCTCCTCCTGAGGCCAATGCATTCCCAACTTCCAGTTCTTTCCTT 1680 ************************************************************ TACCCTGGAGAGTTAGTAAGGTAAGAACCATTCTTTCTCTCCAAAACCACTCCTCCTTGG 1666 TACCCTGGAGAGTTAGTAAGGTAAGAACCATTCTTTCTCTCCAAAACCACTCCTCCTTGG 1740 ************************************************************ CTGGCAAGTTGGTGTCCTAACTCCGTTCTCTTCCTAGCTCATGGCCTCTCTAGATAATAA 1726 CTGGCAAGTTGGTGTCCTAACTCCGTTCTCTTCCTAGCT.CATGGCCTCTCTAGATAATAA 1800 ************************************************************ AGTTGTCTCCTCCTTTCTGGATCTCTTCCTCCTAACACCCCTCCCCTGAAACCCTGGACT 1786 AGTTGTCTCCTCCTTTCTGGATCTCTTCCTCCTAACACCCCTCCCCTGAAACCCTGGACT 1860 ****+******************************+************************ CTGCCCTCTCTCCAAGAAAATCCATCTATTCAACTATTCTTGCATTCAATTACTCTAAAT 1846 CTGCCCTCTCTCCAAGAAAATCCATCTATTCAACTATTCTTGCATTCAATTACTCTAAAT 1920 ****************************************+******************* GAGAGCGTGTTGGAGCTATGGCAAATTCCCTGTTGTCACCTTGCTATTTTGCAGACAACA 1906 GAGAGCGTGTTGGAGCTATGGCAAATTCCCTGTTGTCACCTTGCTATTTTGCAGACAACA 1980 TAATATTTAACCTCTCATAACCAGAGAGGTTAAATAATTTGTCAAATGCAATACAGTAAG 1966 TAATATTTAACCTCTCATAACCAGAGAGGTTAAATAATTTGTCAAATGCAATACAGTAAG 2040 ************************************************************ ACAGAGGCAAGGACAAGGTTTGACTTCCAGCCCAGCCTCTTTTCC'ACAACCTGCTAAATC 2026 ACAGAGGCAAGGACAAGGTTTGACTTCCAGCCCAGCCTCTTTTCCACAACCTGCTAAATC 2100 ************************************************************ CTGATCCATCTGAAAACTTTTCTAATTAGTGAAGATGACTAATAAAAATTTTCCCTATCT 2086 CTGATCCATCTGAAAACTTTTCTAATTAGTGAAGATGACTAATAAAAATTTTCCCTATCT 2160 ************************************************************ CCAAGGTAGGAGCTTTCTGGAAGTTTCTAGAAATTTTCAATAACCACCAGCCAAGGTTAC 2146 CCAAGGTAGGAGCTTTCTGGAAGTTTCTAGAAATTTTCAATAACCACCAGCCAAGGTTAC 2220 ************************************************************ CTCCAGGTAACCTTGCAGCACCAGGCTGGAAGTCAGATCGGCTTCACTATCTTCCAACTC 2206 CTCCAGGTAACCTTGCAGCACCAGGCTGGAAGTCAGATCGGCTTCACTATCTTCCAACTC 2280 ***************** ******************************************* TACAGCCTGTATCTCTCCATCCCCAGCTTTGACCTTTCCTGCTCAAGTAACCTACGGGCA 2266 TACAGCCTGTATCTCTCCATCCCCAGCTTTGACCTTTCCTGCTCAAGTAACCTACGGGCA 2340 ************************************************************ CATCCAGCGTCACTAAAAACTCAGGGCTTTTCTTCCCGGTTACTCCTCCAAGCGTTCCCT 2326 CATCCAGCGTCACTAAAAACTCAGGGCTTTTCTTCCCGGTTACTCCTCCAAGCGTTCCCT 2400 ************************************************************ GGTATCCTCAACCTCAGATCCCAGGTTCAGATTTCTGCAGTCAATCTATGACCCCTCTCT 2386 GGTATCCTCAACCTCAGATCCCAGGTTCAGATTTCTGCAGTCAATCTATGACCCCTCTCT 2460 ************************************************************ TCTTGCATCCTTCATATGCCACCAGACACCATGCCCAGTCCAGCCTGATTTTGAAACAAC ' 24 6 TCTTGCATCCTTCATATGCCACCAGACACCATGCCCAGTCCAGCCTGATTTTGAAACAAC 2520 ************************************************************ TTTCATGCCGGTCTTCTCTTCCCTGACATGTTACTGTCCAGGCTCAAGTCCTCAGCTTCT 2506 TTTCATGCCGGTCTTCTCTTCCCTGACATGTTACTGTCCAGGCTCAAGTCCTCAGCTTCT 2580 ************************************************************ CATATCTGCATCTTTGCAACCAACTTC'CTCCCTTGCCTCTCTGCTTTT.CCATCCCACTTT 2566 CATATCTGCATCTTTGCAACCAACTTCCTCCCTTGCCTCTCTGCTTTTCCATCCCACTTT 2640 ************************************************************ TCATGTGTCCTCCATACCATCTATAACAGTGATCTCCCTGGAACACTCAAGAAGACACAA 2626 TCATGTGTCCTCCATACCATCTATAACAGTGATCTCCCTGGAACACTCAAGAAGACACAA 2700 ************************************************************ CATACC'ATATTATTTAAAGACCAGGGTACTGGACAGTGGCTCACACCTGTATTCCCGACT 2686 CATACCATATTATTTAAAGACCAGGGTACTGGACAGTGGCTCACACCTGTATTCCCGACT 2760 ************************************************************ TTGAGAGTCTGAAGCGGGAGGATCACTTGAGGCCAGGAGTTAAGAGACCAGCCTGGGCAA 2746 TTGAGAGTCTGAAGCGGGAGGATCACTT.GAGGCCAGGAGTTAAGAGACCAGCCTGGGCAA 2820 ************************************************************ CACAGCAAGACCCTGTCTCTAAAAAAAAAAATTAATTAACTGGGTATGGTGGCACATGCC 2806 CACAGCAAGACCCTGTCTCTAAAAAAAAAAATTAATTAACTGGGTATGGTGGCACATGCC 2880 ************************************************************ TGTAGTCCCAGCTACTCAGGAGGCTGAGGTGGGAGGATGACTTGAGCCCAGGAGTTTGAG 2866 TGTAGTCCCAGCTACTCAGGAGGCTGAGGTGGGAGGATGACTTGAGCCCAGGAGTTTGAG 2940 *****************************+****************************** GCTGCAAGGAGCTATGATCATGCCAGTGCATCCCAGCTCTAGGTGAGACAGTGAGATCCG 2926 GCTGCAAGGAGCTATGATCATGCCAGTGCATCCCAGCTCTAGGTGAGACAGTGAGATCCG 3000 ************************************************************ GTCTCCAAAATAAATCAATCAATCAAATAAAGACCAAAGTCAAACCGCAC'ATCAGGATCT 2986 GTCTCCAAAATAAATCAATCAATCAAATAAAGACCAAAGTCAAACCGCACATCAGGATCT 3060 CTCACACCCTTCCAATTTTGCCATCTACCAGCACTTAGCTAAACCCATCTCCCATCTCTT 3046 CTCACACCCTTCCAATTTTGCCATCTACCAGCACTTAGCTAAACCCATCTCCC'ATCTCTT 3120 a*********************************************************** CCACCATGAATTCACTCTTTCAAAAAGGCTAATGTCTTCTTACTCACCCTTGCCTCTAAG 3106 CCACCATGAATTCACTCTTTCAAAAAGGCTAATGTCTTCTTACTCACCCTTGCCTCTAAG 3180 CCTTTGCTATCACCATTTCCCCCAAGCTGGAGGGCCCTCCCTCTCCCTTTACCCCTCTTC 3166 CCTTTGCT AT CACCATTTCCCCC AAGCTGGAGGGCCCT CCCT CT CCCTTT ACCCCTCTT C 3240 CACTACCTCCCACCCCTACTTTTTCCAGAAAGCCATTTCCTCTCTTTTTTCTGATTGATC 3226 CACTACCTCCCACCCCTACTTTTTCCAGAAAGCCATTTCCTCTCTTTTTTCTGATTGATC 3300 **********+**********+*********+*+++*******************+**** CTTCCCTCTCACCCAGGATTAGATGCTGGAAATGACCACTTCTGGAGGGCAGGGAACAAG 3286 CTTCCCTCTCACCCAGGATTAGATGCTGGAAATGACCACTTCTGGAGGGCAGGGAACAAG 3360 *****************************************+************+***** CCCTTAATCTGCATAATGAGTGTTCAATAAACAGTTGTCAAACTTTGAAA 3336 ■ CCCTTAATCTGCATAATGAGTGTTCAATAAACAGTTGTCAAACTTTGAAA 3410 Table XXVB. Nucleotide sequence alignment of 213P1F11 v.l (SEQ. ID. NO.60) and 213P1F11 v.3 (SEQ. ID. NO.61).

CTGACTCATTTAGACTCTCTGCCTAGGCCACCTTTGCCAGAGGGAGTCCCCTCAGCCTTG 60 CTGACTCATTTAGACTCTCTGCCTAGGCCACCTTTGCCAGAGGGAGTCCCCTCAGCCTTG 60 ******************,****************************************** CGATCACTCATCCCATTGGCGTTGGCTCCATTTCCACACCACAGCTGTGTGCCAAGGGTG 120 CGATCACTCATCCCATTGGCGTTGGCTCCATTTCCACACCACAGCTGTGTGCCAAGGGTG 120 ************************************************************ TGTCATGAGGTTTCTTGAGTGACAGAAAACTCACCGACAATAAAGGGCCAGGTGATTGTG 180 TGTCATGAGGTTTCTTGAGTGACAGAAAACTCACCGACAATAAAGGGCCAGGTGATTGTG 180 *****************************************************'******* CCACCCGATTCATAGACCAGGCTTCTCAGGAGAAACCCCGGGAGATTCCACACTGTCAGC 240 CCACCCGATTCATAGACCAGGCTTCTCAGGAGAAACCCCGGGAGATTCCACACTGTCAGC 240 ************************************************************ CCCTTCTCCAAGATCAGTACGTGGGCCTGACTCCTCCTCGGTGCCCAGCTCAGTATTGGC 300 CCCTTCTCCAAGATCAGTACGTGGGCCTGACTCCTCCTCGGTGCCCAGCTCAGTATTGGC 300 ************************************************************ AACTAGGAGAGTAGTGAGATTGAACTTGGCCTTGAGGAACAGCTGCCTCTAGAGTTGGAT 360 AACTAGGAGAGTAGTGAGATTGAACTTGGCCTTGAGGAACAGCTGCCTCTAGAGTTGGAT 360 ************************************************************ CAGACAAGGGTGCTGAGAGCCGGGACTCACAACCAAAGGAGAAATGAGCAATCCGCGGTC 420 CAGACAAGGGTGCTGAGAGCCGGGACTCACAACCAAAGGAGAAATGAGCAATCCGGGGTC 420 ************************************************************ TTTGGAAGAGGAGAAATATGATATGTCAGGTGCCCGCCTGGCCCTAATACTGTG.TGTCAC 480 TTTGGAAGAGGAGAAATATGATATGTCAGGTGCCCGCCTGGCCCTAATACTGTGTGTCAC 480' ************************************************************ CAAAGCCCGGGAAGGTTCCGAAGAAGACCTGGATGCTCTGGAACACATGTTTCGGCAGCT 540 . CAAAGCCCGGGAAGGTTCCGAAGAAGACCTGGATGCTCTGGAACACATGTTTCGGCAGCT 540 ************************************************************ GAGATTCGAAAGCACCATGAAAAGAGACCCCACTGCCGAGCAATTCCAGGAAGAGCTGGA 600 GAGATTCGAAAGCACCATGAAAAGAGACCCCACTGCCGAGCAATTCCAGGAAGAGCTGGA 600 ************************************************************ AAAATTCCAGCAGGCCATCGATTCCCGGGAAGATCCCGTCAGTTGTGCCTTCGTGGTACT 660 AAAATTCCAGCAGGCCATCGATTCCCGGGAAGATCCCGTCAGTTGTGCCTTCGTGGTACT 660 ************************************************************ CAT GGCT CACGGGAGGGAAGGCTT CCTCAAGGGAGAAGATGGGGAGAT GGTCAAGCTGGA 720 CAT GGCT CACGGGAGGGAAGGCTT CCTCAAGGGAGAAGATGGGGAGAT GGTCAAGCTGGA 720 ************************************************************ GAATCTCTTCGAGGCCCTGAACAACAAGAACTGCCAGGCCCTGCGAGCTAAGCCCAAGGT 780 GAATCTCTTCGAGGCCCTGAACAACAAGAACTGCCAGGCCCTGCGAGCTAAGCCCAAGGT 780 GTACATCATACAGGCCTGTCGAGGAG 806 GTACATCATACAGGCCTGTCGAGGAGCCACCCTGCCCAGCCCCTTTCCTTACCTTTCTCT ' 840 ************************** ■ AACAAAGGGACCCCGGTGAAACAGTA 832 CTGACTTTGCCTCCTCCTCTTCTTGTTGTTTCAGAACAAAGGGACCCCGGTGAAACAGTA 900 213P1F11V.1 GGTGGAGATGAGATTGTGATGGTCATCAAAGACAGCCCACAAACCATCCCAACATACACA 892 213P1 Fl 1 v .3 GGTGGAGATGAGATTGTGATGGTCATCAAAGACAGCCCACAAACCATCCCAACATACACA 960 ************************************************************ 213P1F11V.1 GATGCCTTGCACGTTTATTCCACGGTAGAGGGATACATCGCCTACCGACATGATCAGAAA 952 213P1 Fl 1v .3 GATGCCTTGCACGTTTATTCCACGGTAGAGGGATACATCGCCTACCGACATGATCAGAAA 1020 ************************************************************ 213P1F11V.1 GGCTCATGCTTTATCCAGACCCTGGTGGATGTGTTCACGAAGAGGAAAGGACATATCTTG 1012 213PlFllv;3 GGCTCATGCTTTATCCAGACCCTGGTGGATGTGTTCACGAAGAGGAAAGGACATATCTTG 1080 ************************************************************ 213P1F11V.1 GAACTTCTGACAGAGGTGACCCGGCGGATGGCAGAAGCAGAGCTGGTTCAAGAAGGAAAA 1072 213PI Fl 1v .3 GAACTTCTGACAGAGGTGACCCGGCGGATGGCAGAAGCAGAGCTGGTTCAAGAAGGAAAA 1140 213P1 Fl 1v .1 GCAAGGAAAACGAACCCTGAAATCCAAAGCACCCT.CCGGAAACGGCTGTATCTGCAGTAG 1132 213P1F11V.3 GCAAGGAAAACGAACCCTGAAATCCAAAGCACCCTCCGGAAACGGCTGTATCTGCAGTAG 1200 ************************************************************ 213PI Fl 1v .1 AAGTAGAAAGACCAGGAGGAGCTTTCCTTCCAGCATTCTTTCTGTCTCACAGAAATTTAG 1192 213P1 11V.3 AAGTAGAAAGACCAGGAGGAGCTTTCCTTCCAGCATTCTTTCTGTCTCACAGAAATTTAG 1260 ************************************************************ 213P1F11V.1 AGGCAGCTCTTACCTCTCCCCAAGATCTTCTGTTCCCAAGGCCAAATGGCACCCAGTTTC 1252 213P1 11V.3 AGGCAGCTCTTACCTCTCCCCAAGATCTTCTGTTCCCAAGGCCAAATGGCACCCAGTTTC 1320 *************************** *.* ******************************* 213P1F11V.1 TTTTCCATCACACCCTTCATGCAGGTCCTCCTGTCCTTATTAGAGCAAGCCAGCCAAAAC 1312 213P1F11V.3 TTTTCCATCACACCCTTCATGCAGGTCCTCCTGTCCTTATTAGAGCAAGCCAGCCAAAAC 1380 ************************************************************ 213P1F11V.1 TTAGCACAAGGCATGGTGGCAACATTAACATCACCTCCCTCAGGCTGGACTTTCTATCTT 1312 213P1F11V.3 TTAGCACAAGGCATGGTGGCAACATTAACATCACCTCCCTCAGGCTGGACTTTCTATCTT 1440 ************************************************************ 213PI Fl 1v .1 TATTAATGCAACCGAAGAGACCTAAGAGTGCATTCACTTATCCCACTTTCTGTTCCTGTG 1432 213P1 Fl 1v .3 TATTAATGCAACCGAAGAGACCTAAGAGTGCATTCACTTATCCCACTTTCTGTTCCTGTG 1500 '■ ************************************************************ 213P1F11V.1 GTCTTCTTTCTCCCATGAAGCAGAAACTGGATAAAGCTCAAGATTTTCCATAGACAAACC 1492 213PI Fl 1v .3 GTCTTCTTTCTCCCATGAAGCAGAAACTGGATAAAGCTCAAGATTTTCCATAGACAAACC 1560 ************************************+*********************** 213P1F11V.1 AAAGCCCACTCATCCCCTCCTACCCCAATCCAACCTCTGCTGGCTCCTGCATCTCACTTG 1552 21 PI Fl 1v .3 AAAGCCCACTCATCCCCTCCTACCCCAATCCAACCTCTGCTGGCTCCTGCATCTCACTTG 1620 ******************************************************* 213P1F11V .1 GAGGTCAAACCTCCTCCTGAGGCCAATGCATTCCCAACTTCCAGTTCTTTCCTTTACCCT 1612 .213P1F11V.3 GAGGTCAAACC CCTCCTGAGGCCAATGCATTCCCAACTTCCAGTTCTTTCCTTTACCCT 1680 ******************************************************+***** j 213P1F11V.1 GGAGAGTTAGTAAGGTAAGAACCATTCTTTCTCTCCAAAACCACTCCTCCTTGGCTGGCA 1672 213P1F11V.3 GGAGAGTTAGTAAGGTAAGAACCATTCTTTCTCTCCAAAACCACTCCTCCTTGGCTGGCA 1740 ************************************************************ 213P1F11V.1 AGTTGGTGTCCTAACTCCGTTCTCTTCCTAGCTCATGGCCTCTCTAGATAATAAAGTTGT 1732 213P1 Fl 1 .3 AGTTGGTGTCCTAACTCCGTTCTCTTCCTAGCTCATGGCCTCTCTAGATAATAAAGTTGT 1800 ************************************************************ 213P1F11V.1 CTCCTCCTTTCTGGATCTCTTCCTCCTAACACCCCTCCCCTGAAACCCTGGACTCTGCCC 1792 213P1F11V.3 CTCCTCCTTTCTGGATCTCTTCCTCCTAACACCCCTCCCCTGAAACCCTGGACTCTGCCC 1860 ************************************************************ 213P1F11V.1 TCTCTCCAAGAAAATCCATCTATTCAACTATTCTTGCATTCAATTACTCTAAATGAGAGC 1852 213P1 Fl 1v .3 TCTCTCCAAGAAAATCCATCTATTCAACTATTCTTGCATTCAATTACTCTAAATGAGAGC 1920 ************************************************************ 213P1F11V.1 GTGTTGGAGCTATGGCAAATTCCCTGTTGTCACCTTGCTATTTTGCAGACAACATAATAT 1912 213P1F11V.3- GTGTTGGAGCTATGGCAAATTCCCTGTTGTCACCTTGCTATTTTGCAGACAACATAATAT 1980 ****************************+***+*+******+****************** TTAACCTCTCATAACCAGAGAGGTTAAATAATTTGTCAAATGCAATACAGTAAGACAGAG 1972 TTAACCTCTCATAACCAGAGAGGTTAAATAATTTGTCAAATGCAATACAGTAAGACAGAG 2040 *************'**************************** + ****************** GCAAGGACAAGGTTTGACTTCCAGCCCAGCCTCTTTTCCACAACCTGCTAAATCCTGATC 2032 GCAAGGACAAGGTTTGACTTCCAGCCCAGCCTCTTTTCCACAACCTGCTAAATCCTGATC 2100 **************************** ******************************** CATCTGAAAACTTTTCTAATTAGTGAAGATGACTAATAAAAATTTTCCCTATCTCCAAGG 2092 CAT C GAAAACTTTTCTAATTAGT GAAGATGACTAAT AAAAATTTTCCCT A CT CCAAGG 2160 *********'*************************************************** TAGGAGCTTTCTGGAAGTTTCTAGAAATTTTCAATAACCACCAGCCAAGGTTACCTCCAG 2152 TAGGAGCTTTCTGGAAGTTTCTAGAAATTTTCAATAACCACCAGCCAAGGTTACCTCCAG 2220 ************************************************************ GTAACCTTGCAGCACCAGGCTGGAAGTCAGATCGGCTTCACTATCTTCCAACTCTACAGC 2212 GTAACCTTGCAGCACCAGGCTGGAAGTC AGATCGGCTTCACTATCTTCCAACTCTACAGC 2280 ***++******************************************************* CTGTATCTCTCCATCCCCAGCTTTGACCTTTCCTGCTCAAGTAACCTACGGGCACATCCA 2272 CTGTATCTCTCCATCCCCAGCTTTGACCTTTCCTGCTCAAGTAACCTACGGGCACATCCA 2340 ************************************************************ GCGTCACTAAAAACTCAGGGCTTTTCTTCCCGGTTACTCCTCCAAGCGTTCCCTGGTATC 2332 GCGTCACTAAAAACTCAGGGCTTTTCTTCCCGGTTACTCCTCCAAGCGTTCCCTGGTATC 2400 **************************************+************+******** CTCAACCTCAGATCCCAGGTTCAGATTTCTGCAGTCAATCTATGACCCCTCTCTTCTTGC 2392 CTCAACCTCAGATCCCAGGTTCAGATTT CTGCAGTCAATCTATGACCCCTCTCTTCTTGC 2460 ******************************************** ATCCTTCATATGCCACCAGACACCATGCCCAGTCCAGCCTGATTTTGAAACAACTTTCAT 2452 ATCCTTCATATGCCACCAGACACCATGCCCAGTCCAGCCTGATTTTGAAACAACTTTCAT 2520 ************************************************************ GCCGGTCTTCTCTTCCCTGACATGTTACTGTCCAGGCTCAAGTCCTCAGCTTCTCATATC 2512 GCCGGTCTTCTCTTCCCTGACATGTTACTGTCCAGGCTCAAGTCCTCAGCTTCTCATATC 2580 ************************************************************ TGCATCTTTGCAACCAACTTCCTCCCTTGCCTCTCTGCTTTTCCATCCCACTTTTCATGT 2572 TGCATCTTTGCAACCAACTTCCTCCCTTGCCTCTCTGGTTTTCCATCCCACTTTTCATGT 2640 ************************************************************ GTCCTCCATACCATCTATAACAGTGATCTCCCTGGAACACTCAAGAAGACACAACATACC 2632 GTCCTCCATACCATCTATAACAGTGATCTCCCTGGAACACTCAAGAAGACACAACATACC 2700 ************************************************************ ATATTATTTAAAGACCAGGGTACTGGACAGTGGCTCACACCTGTATTCCCGACTTTGAGA 2692 ATATTATTTAAAGACCAGGGTACTGGACAGTGGCTCACACCTGTATTCCCGACTTTGAGA 2760 ***********************************************+********+*** GTCTGAAGCGGGAGGATCACTTGAGGCCAGGAGTTAAGAGACCAGCCTGGGCAACACAGC 2752 GTCTGAAGCGGGAGGATCACTTGAGGCCAGGAGTTAAGAGACCAGCCTGGGCAACACAGC 2820 ************************************************************ AAGACCCTGTCTCTAAAAAAAAAAATTAATTAACTGGGTATGGTGGCACATGCCTGTAGT 2812 AAGACCCTGTCTCTAAAAAAAAAAATTAATTAACTGGGTATGGTGGCACATGCCTGTAGT 2880 ************************************************************ CCCAGCTACTCAGGAGGCTGAGGTGGGAGGATGACTTGAGCCCAGGAGTTTGAGGCTGCA 2872 CCCAGCTACTCAGGAGGCTGAGGTGGGAGGATGACTTGAGCCCAGGAGTTTGAGGCTGCA 2940 ************************************************************ AGGAGCTATGATCATGCCAGTGCATCCCAGCTCTAGGTGAGACAGTGAGATCCGGTCTCC 2932 AGGAGCTATGATCATGCCAGTGCATCCCAGCTCTAGGTGAGACAGTGAGATCCGGTCTCC 3000 ************************************************************ AAAATAAATCAATCAATCAAATAAAGACCAAAGTCAAACCGCACATCAGGATCTCTCACA 2992 213P1 Fl 1v .3 AAAATAAATCAATCAATCAAATAAAGACCAAAGTCAAACCGCACATCAGGATCTCTCACA 3060 ************************************************************ 213P1F1 lv.1 CCCTTCCAATTTTGCCATCTACCAGCACTTAGCTAAACCCATCTCCCATCTCTTCCACCA 3052 213P1F11V.3 CCCTTCCAATTTTGCCATCTACCAGCACTTAGCTAAACCCATCTCCCATCTCTTCCACCA 3120 ************************************************************ 213P1F11V.1 TGAATTCACTCTTTCAAAAAGGCTAATGTCTTCTTACTCACCCTTGCCTCTAAGCCTTTG 3112 213P1 Fl 1v .3 TGAATTCACTCTTTCAAAAAGGCTAATGTCTTCTTACTCACCCTTGCCTCTAAGCCTTTG 3180 ************************************************************ 213P1F11V.1 CTATCACCATTTCCCCCAAGCTGGAGGGCCCTCCCTCTCCCTTTACCCCTCTTCCACTAC 3172 213P1F11V.3 CTATCACCATTTCCCCCAAGCTGGAGGGCCCTCCCTCTCCCTTTACCCCTCTTCCACTAC 3240 ************************************************************ 213P1F11V.1 CTCCCACCCCTACTTTTTCCAGAAAGCCATTTCCTCTCTTTTTTCTGATTGATCCTTCCC 3232 213P1 11V.3 CTCCCACCCCTACTTTTTCCAGAAAGCCATTTCCTCTCTTTTTTCTGATTGATCCTTCCC 3300 ************************************************************ 213P1F11V.1 TCTCACCCAGGATTAGATGCTGGAAATGACCACTTCTGGAGGGCAGGGAACAAGCCCTTA 3292 213P1F11V.3 TCTCACCCAGGATTAGATGCTGGAAATGACCACTTCTGGAGGGCAGGGAACAAGCCCTTA 3360 ************************************************************ 213P1F11V.1 ATCTGCATAATGAGTGTTCAATAAACAGTTGTCAAACTTTGAAA 3336 213P1F11V.3 ATCTGCATAATGAGTGTTCAATAAACAGTTGTCAAACTTTGAAA 3404 ******************************************** Table XXVC. Nucleotide sequence alignment of 213P1F11 v. l (SEQ. ID. NO. 62) and 213P1F11 v.4 (SEQ ID. NO. 63). 213PI Fl 1 .1 CTGACTCATTTAGACTCTCTGCCTAGGCCACCTTTGCCAGAGGGAGTCCCCTCAGCCTTG 60 213P1F11V.4 213P1Fl 1v .1 CGATCACTCATCCCATTGGCGTTGGCTCCATTTCCACACCACAGCTGTGTGCCAAGGGTG .120 213P1F11V.4 213PI Fl 1v .1 TGTCATGAGGTTTCTTGAGTGACAGAAAACTCACCGACAATAAAGGGCCAGG-TGATTGT 1 9 213P1F11V.4 ATGGGGAAAT-GCCAAGAGTATGACAAAAGTCTGTCTGT 38 * * * * * ** * * ** ** ** *** 213P1F11V.1 GCCACCCGATTCATAGACCAGGCTTCTCAGGAGAAACCCCGGGAGATTCCACACTGTCAG 239 213P1F1 lv .4 GCAGCCAGAGA-AGAGAACAGGACTCAGAGATGAGAAT GGAGAATGTGGACAGACA- 93 * * * * * * * *** **** ** ** ** * ***** * ** * ** 213P1F1 lv .1 CCCCTTCTCCAAGATCAGTACGTGGGCCTGACTCCTCCTCGGTGCCCAGCTCAGTATTGG 299 213P1 Fl 1v .4 TTCAGACTCAAGGAAGAGCAAGGGAG GGCTTTCAGGGGAAGTTCAGTCCACCAGAAG 150 * +** * ** ** * * * * * ** * * *** ** * * 213P1F11V.1 CAACTAGGAGAGTAGTGAGATTGAACTTGGCCTTGAGGAACAGCTGCCTCTAGAGTTGGA 359 213P1 Fl 1v .4 CTGGTGAATGACCCACGGGA---GACAC AGGAA GTTTTTGGGGGCGGA 195 * * ** * ** ** ***** * * * * * *** 213P1 Fl lv .1 TCAGACAAGGGTGCTGAGAGCCGGGA-CTCACAACCAAAGG-AGAAATGAGCAATCCGCG 17 213P1 Fl 1v .4 GTGGG GGACATTG GGGACGGGATCTC'AGTATTAGCTTCAGAAACTCTGAGACCTCT 252 * ** ** * * ***** **** * * ***** * ** * 213P1F1 lv.1 GTCTTTGGAAGAGGAGAAATATGATATGTCAGGTGCCCGCCTGGCCCTAATACTGTGTGT 477 213P1F11V.4 GCAAGTG-AGGAGGAGAAATATGATATGTCAGGTGCCCGCCTGGCCCTAATACTGTGTGT 311 * ** * ************************************************** 213P1F11V.1 CACCAAAGCCCGGGAAGGTTCCGAAGAAGACCTGGATGCTCTGGAACACATGTTTCGGCA 537 213P1F1 lv.4 CACCAAAGCCCGGGAAGGTTCCGAAGAAGACCTGGATGCTCTGGAACACATGTTTCGGCA 371 ************************************************************ 213P1F11V.1 GCTGAGATTCGAAAGCACCATGAAAAGAGACCCCACTGCCGAGCAATTCCAGGAAGAGCT 597 213PI Fl 1v .4 GCTGAGATTCGAAAGCACCATGAAAAGAGACCCCACTGCCGAGCAATTCCAGGAAGAGCT 431 ***********************++********+**********+*************** 213P1F11V.1 GGAAAAATTCCAGCAGGCCATCGATTCCCGGGAAGATCCCGTCAGTTGTGCCTTCGTGGT 657 213P1 Fl 1v .4 GGAAAAATTCCAGCAGGCCATCGATTCCCGGGAAGATCCCGTCAGTTGTGCCTTCGTGGT 491 ************************************************************ 213P1F11V.1 ACTCATGGCTCACGGGAGGGAAGGCTTCCTCAAGGGAGAAGATGGGGAGATGGTCAAGCT 717 213P1 Fl 1v .4 ACTCATGGCTCACGGGAGGGAAGGCTTCCTCAAGGGAGAAGATGGGGAGATGGTCAAGCT 551 ************************************************************ 213P1F11V.1. GGAGAATCTCTTCGAGGCCCTGAACAACAAGAACTGCCAGGCCCTGCGAGCTAAGCCCAA 777 213P1F1 lv .4 GGAGAATCTCTTCGAGGCCCTGAACAACAAGAACTGCCAGGCCCTGCGAGCTAAGCCCAA 611 ************************************************************ 213P1F11V.1 GGTGTACATCATACAGGCCTGTCGAGGAGAACAAAGGGACCCCGGTGAAACAGTAGGTGG 837 213P1 Fl 1v .4 GG GTACATCATACAGGCCTGTCGAGGAGAACAAAGGGACCCCGGTGAAACAGTAGGTGG 671 ************************************************************ 213P1F11V.1 AGATGAGATTGTGATGGTCATCAAAGACAGCCCACAAACCATCCCAACATACACAGATGC 897 213P1 Fl 1v .4 AGATGAGATTGTGATGGTCATCAAAGACAGCCCACAAACCATCCCAACATACACAGATGC 731 ************************************************************ 213P1F11V.1 CTTGCACGTTTATTCCACGGTAGAGGGATACATCGCCTACCGACATGATCAGAAAGGCTC 957 213P1F11V.4 CTTGCACGTTTATTCCACGGTAGAGGGATACATCGCCTACCGACATGATCAGAAAGGCTC 791 ************************************************************ 213P1F11V.1 ATGCTTTATCCAGACCCTGGTGGATGTGTTCACGAAGAGGAAAGGACATATCTTGGAACT 1017 213P1F11V.4 ATGCTTTATCCAGACCCTGGTGGATGTGTTCACGAAGAGGAAAGGACATATCTTGGAACT 851 213P1 Fl 1v .1 TCTGACAGAGGTGACCCGGCGGATGGCAGAAGCAGAGCTGGTTCAAGAAGGAAAAGCAAG 1077 213P1 Fl 1v .4 TCTGACAGAGGTGACCCGGCGGATGGCAGAAGCAGAGCTGGTTCAAGAAGGAAAAGCAAG 911 ************************************************************ , 213P1F11V.1 GAAAACGAACCCTGAAATCCAAAGCACCCTCCGGAAACGGCTGTATCTGCAGTAGAAGTA 1137 213P1 Fl 1 .4 GAAAACGAACCCTGAAATCCAAAGCACCCTCCGGAAACGGCTGTATCTGCAGTAG 966 ******************************************************* 213PI Fl 1 .1 GAAAGACCAGGAGGAGCTTTCCTTCCAGCATTCTTTCTGTCTCACAGAAATTTAGAGGCA 1197 213P1F11V.4 213P1F11V.1 GCTCTTACCTCTCCCCAAGATCTTCTGTTCCCAAGGCCAAATGGCACCCAGTTTCTTTTC 1257 213P1F11V.4 213P1F11V.1 CATCACACCCTTCATGCAGGTCCTCCTGTCCTTATTAGAGCAAGCCAGCCAAAACTTAGC 1317 213P1F11V.4 . 213P1 11V.1 ACAAGGCATGGTGGCAACATTAACATCACCTCCCTCAGGCTGGACTTTCTATCTTTATTA 1377 213P1F11V.4 213P1F11V.1 ATGCAACCGAAGAGACCTAAGAGTGCATTCACTTATCCCACTTTCTGTTCCTGTGGTCTT 1 37 213P1F11V.4 213P1F11V.1 CTTTCTCCCATGAAGCAGAAACTGGATAAAGCTCAAGATTTTCCATAGACAAACCAAAGC 1497 213P1F11V.4 213PI Fl 1v .1 CCACTCATCCCCTCCTACCCCAATCCAACCTCTGCTGGCTCCTGCATCTCACTTGGAGGT 1557 213P1F11V.4 ■ 213P1F11V.1 CAAACCTCCTCCTGAGGCCAATGCATTCCCAACTTCCAGTTCTTTCCTTTACCCTGGAGA 1617 213P1F11V.4 ' 213P1F11V.1 GTTAGTAAGGTAAGAACCATTCTTTCTCTCCAAAACCACTCCTCCTTGGCTGGCAAGTTG 1677 213P1F11V.4 213P1F11V.1 GTGTCCTAACTCCGTTCTCTTCCTAGCTCATGGCCTCTCTAGATAATAAAGTTGTCTCCT 1737 213P1F11V.4 ' 213 PI Fl 1v .1 CCTTTCTGGATCTCTTCCTCCTAACACCCCTCCCCTGAAACCCTGGACTCTGCCCTCTCT 1797 213P1F11V.4 ' 213P1F11V.1 CCAAGAAAATCCATCTATTCAACTATTCTTGCATTCAATTACTCTAAATGAGAGCGTGTT 1857 213P1F11V.4 : ' 213P1F1 lv .1 GGAGCTATGGCAAATTCCCTGTTGTCACCTTGCTATTTTGCAGACAACATAATATTTAAC 1917 213P1F11V.4 — 213PI Fl 1 .1 CTCTCATAACCAGAGAGGTTAAATAATTTGTCAAATGCAATACAGTAAGACAGAGGCAAG 1977 213P1F11V.4 : 213P1F1 lv.1 GACAAGGTTTGACTTCCAGCCCAGCCTCTTTTCCACAACCTGCTAAATCCTGATCCATCT 2037 213P1F11V.4 213P1F11V.1 GAAAACTTTTCTAATTAGTGAAGATGACTAATAAAAATTTTCCCTATCTCCAAGGTAGGA 2097 213P1F11V.4 : 213P1F11V.1 GCTTTCTGGAAGTTTCTAGAAATTTTCAATAACCACCAGCCAAGGTTACCTCCAGGTAAC 2157 213P1F11V.4 213P1F1 lv .1 CTTGCAGCACCAGGCTGGAAGTCAGATCGGCTTCACTATCTTCCAACTCTACAGCCTGTA 2217 213PlFllv.'4 213P1F11V.1 TCTCTCCATCCCCAGCTTTGACCTTTCCTGCTCAAGTAACCTACGGGCACATCCAGCGTC 2277 213P1F11V.4 213P1F1 lv.1 ACTAAAAACTCAGGGCTTTTCTTCCCGGTTACTCCTCCAAGCGTTCCCTGGTATCCTCAA 2337 213P1F11V.4 213P1F11V.1 CCTCAGATCCCAGGTTCAGATTTCTGCAGTCAATCTATGACCCCTCTCTTCTTGCATCCT 2397 213P1F11V.4 213 PI Fl 1 .1 TCATATGCCACCAGACACCATGCCCAGTCCAGCCTGATTTTGAAACAACTTTCATGCCGG 2457 213P1F11V.4 ' 213P1F11V.1 TCTTCTCTTCCC'TGACATGTTACTGTCCAGGCTCAAGTCCTCAGCTTCTCATATCTGCAT 2517 213P1F11V.4 213 P1 Fl 1v .1 CTTTGCAACCAACTTCCTCCCTTGCCTCTCTGCTTTTCCATCCCACTTTTCATGTGTCCT 2577 213PI Fl 1v .1 CCATACCATCTATAACAGTGATCTCCCTGGAACACTCAAGAAGACACAACATACCATATT 2637 213P1F11V.4 213P1F11V.1 ATTTAAAGACCAGGGTACTGGACAGTGGCTCACACCTGTATTCCCGACTTTGAGAGTCTG 2697 213P1F11V.4 213P1F11.V.1 AAGCGGGAGGATCACTTGAGGCCAGGAGTTAAGAGACCAGCCTGGGCAACACAGCAAGAC 2757 213P1F11V.4 213P1F11V.1 CCTGTCTCTAAAAAAAAAAATTAATTAACTGGGTATGGTGGCACATGCCTGTAGTCCCAG 2817 213P1F11V.4 213P1 l 1 v .1 CTACTCAGGAGGCTGAGGTGGGAGGATGACTTGAGCCCAGGAGTTTGAGGCTGCAAGGAG 2877 213P1F11V.4 213P1F11V.1 CTATGATCATGCCAGTGCATCCCAGCTCTAGGTGAGACAGTGAGATCCGGTCTCCAAAAT 2937 213P1F11V.4 213P1F11V.1 AAATC'AATCAATCAAATAAAGACCAAAGTCAAACCGCACATCAGGATCTCTCACACCC'TT 2997 213P1F11V.4 213P1F11V. CCAATTTTGCCATCTACCAGCACTTAGCTAAACCCATCTCCCATCTCTTCCACCATGAAT 3057 213P1F11V.4 TCACTCTTTCAAAAAGGCTAATGTCTTCTTACTCACCCTTGCCTCTAAGCCTTTGCTATC 3117 ACCATTTCCCCCAAGCTGGAGGGCCCTCCCTCTCCCTTTACCCCTCTTCCAC-TACCTCCC 3177 ACCCCTACTTTTTCCAGAAAGCCATTTCCTCTCTTTTTTCTGATTGATCCTTCCCTCTCA 3237 CCCAGGATTAGATGCTGGAAATGACCACTTCTGGAGGGCAGGGAACAAGCCCTTAATCTG 3297 CATAATGAGTGTTCAATAAACAGTTGTCAAACTTTGAAA 3336 1 Table XXVIA. Peptide sequences of protein coded by 213P1F11 v.2 (SEQ. ID. NO.64).

MSNPRSLEEE KYDMSGARLA LILCVTKARE GSEEDLDALE HMFRQLRFES TMKRDPTAEQ 60 FQEELEKFQQ AIDSREDPVS CAFWLMAHG REGFLKGEDG EMVKLENLFE ALNN NCQAL 120 RAKPKVY I IQ ACRGEQRDPG ETVGGDEIVM VIKDSPQTIP TYTDALHVYS TVEGPTPFQD 180 PLYLPSEAPP NPPLWNSQDT SPTDMI RKAH ALSRPWWMCS RRGKDISWNF 230 Table XXVIB. Peptide sequences of protein coded by 213P1F11 v.3 (SEQ. ID. NO.65).

MSNPRSLEEE KYDMSGARLA LILCVTKARE GSEEDLDALE HMFRQLRFES TMKRDPTAEQ 60 FQEELEKFQQ AIDSREDPVS CAFWLMAHG REGFLKGEDG EMVKLENLFE ALNNKNCQAL 120 RAKPKVY HQ ACRGATLPSP FPYLSL 146 Table XXVI C. Peptide sequences of protein coded by 213P1F11 v.4 (SEQ. ID. NO.66) MGKCQEYDKS LSVQPEKRTG LRDENGECGQ TFRLKEEQGR AFRGSSVHQK LVNDPRETQE 60 VFGGGVGDIV GRDLSISFRN SETSASEEEK YDMSGARLAL ILCVTKAREG SEEDLDALEH 120 MFRQLRFEST MKRDPTAEQF QEELEKFQQA IDSREDPVSC AFWLMAHGR EGFLKGEDGE 180 MVKLENLFEA LNNKNCQALR AKPKVYIIQA CRGEQRDPGE TVGGDEIVMV IKDSPQTIPT 240 YTDALHVYST VEGYIAYRHD QKGSCFIQTL VDVFTKRKGH I LELLTEVTR RMAEAELVQE 300 GKARKTNPEI QSTLRKRLYL Q 321 Table XXVIIA. Amino acid sequence alignment of 213P1F11 v. l (SEQ. ID. NO. 67) and 213P1F11 v.2 (SEQ. ID. NO. 68) 213P1F11V.1 MS PRSLEEEKYDMSGARLALI LCVTKAREGSEEDLDALEHMFRQLRFESTMKRDPTAEQ 60 213P1FHV.2 MSNPRSLEEEKYDMSGARLALI LCVTKAREGSEEDLDALEHMFRQLRFESTMKRDPTAEQ- 60 ************************************************************ 213P1F1 lv.1 FQEELEKFQQAIDSREDPVSCAFVVLMAHGREGFLKGEDGEMVKLENLFEALNNKNCQAL 120 213P1F1 lv.2 FQEELEKFQQAIDSREDPVSCAFVVLMAHGREGFLKGEDGEMVKLENLFEALNNKNCQAL 120 2 1 3 P 1 F1 1 V . 1 RA P VY I IQACRGEQRDPGET GGDEIVMVIKDS PQTI PT TDALHVYSTVEGYIAYRH 180 2 1 3 P1 1 1 V . 2 RAKPKVY I IQACRGEQRDPGETVGGDEI MVIKDSPQTI PTYTDALHVYSTVEGPTPFQD 180 ****************************************************** - · 213P1F11V.1 DQKGSCFIQTLVDVFTKRKGHI LELLTEVTRRMAEAELVQEGKARKTNPEIQSTLRKRLY 240 213P1F11V.2 PLYLPSEAPPNPPLWN SQDTSPTDMIRKAHALSRPWWMCSRRGKDIS 227 Table XXVIIB. Amino acid sequence alignment of 213P1F11 v.l (SEQ. ID. NO. 69) and 213P1F11 v (SEQ. ID. NO. 70) 213P1F1 lv.1 MSNPRSLEEEKYDMSGARLALI LCVTKAREGSEEDLDALEHMFRQLRFESTMKRDPTAEQ 60 213P1F1 lv.3 MSNPRSLEEEKYDMSGARLALI LCVTKAREGSEEDLDALEHMFRQLRFESTMKRDPTAEQ 60 ************************************************************ 213P1F11V.1 FQEELEKFQQAIDSREDPVSCAFVVLMAHGREGFLKGEDGEMVKLENLFEALNNKNCQAL 120 213P1F11V.3 FQEELEKFQQAIDSREDPVSCAFWLMAHGREGFLKGEDGEMVKLENLFEALNNKNCQAL 120 *******+*********************************+*********+******** 213P1F1 lv . '1 RAKPKVY I IQACRGEQRDPGETVGGDEIVMVIKDSPQTI PTYTDALHVYSTVEGYIAYRH 180 213P1F11V.3 RAKPKVYI IQACRG ATLPSPFPYLSL 146 213P1 Fl lv .1 DQKGSCFIQTLVDVFTKRKGHI LELLTEVTRRMAEAELVQEGKARKTNPEIQSTLRKRLY 240 213P1F11V. 3 ' --' 213P1F11V.1 LQ 242 213P1F11V. 3 - - ■ .

Table XXVllC. Amino acid sequence alignment of 213P1F11 v.l (SEQ. ID. NO. 71) and 213P1F11 v (SEQ. ID. NO. 72). 213P1F11V. 1 MSNPR 5 213PlFllv.'3 MGKCQEYDKSLSVQPEKRTGLRDENGECGQTFRLKEEQGRAFRGSSVHQKLVNDPRETQE 60 : . : * * 213P1F11V. 1 SLEEEKYDMSGARLALILCVTKAREGSEEDLDALEH 41 213P1F11V.3 VFGGGVGDIVGRDLSI SFRNSETSASEEEKYDMSGARLALI LCVTKAREGSEEDLDALEH 120 ■ ********************************** 213P1F11V. 1 MFRQLRFESTMKRDPTAEQFQEELEKFQQAI DSREDPVSCAFVVLMAHGREGFLKGEDGE 101 2I3P1F11V.3 MFRQLRFESTMKRDPTAEQFQEELEKFQQAI DSREDPVSCAFVVLMAHGREGFLKGEDGE 180 213P1F11V.1 . MVKLENLFEALNNKNCQALRAKPKVY11QACRGEQRDPGETVGGDEIVMVIKDSPQT I PT 161 213P1F1 lv.3 MVKLENLFEALNNKNCQALRAKPKVYI IQACRGEQRDPGETVGGDEIVMVIKDSPQ I PT 240 213P1F11V. 1 YTDALHVYSTVEGYIAYRHDQKGSCFIQTLVDVFTKRKGHI LELLTEVTRRMAEAELVQE 221 213P1F11V.3 YTDALHVYSTVEGYIAYRHDQKGSCFIQTLVDVFTKRKGHI LELLTEVTRRMAEAELVQE 300 ************************************************************ 213P1F11V.1 GKARKTNPEIQSTLRKRLYLQ 242 213PIF11V.3 GKARKTNPEIQSTLRKRLYLQ 321 ********************* Table XXVIII. Clustal Alignment of 213P1F11 protein variants. (SEQ. ID. NOs: ) v. l_ v.2_ v.3_ v . _ MGKCQEYDKSLS VQPEKRTGLRDENGECGQTFRLKEEQGRAFRGSSVHQKLVNDPRETQE v.5_ ■ v.6_ v.1_ MSNPRSLEEEKYDMSGARLALILCVTKAREGSEEDLDALEH v .2_ MSNPRSLEEEKYDMSGARLALILCVTKAREGSEEDLDALEH v.3_ MSNPRSLEEE YDMSGARLALILCVTKAREGSEEDLDALEH v. _ VFGGGVGDIVGRDLSISFRNSETSASEEEKYDMSGARLALILCVTKAREGSEEDLDALEH v .5_ MSNPRSLEEE YDMSGARLALILRVTKAREGSEEDLDALEH v.6_ — MSNPRSLEEEKYDMSGARLALILCVTKAREGSEEDLDALEH * ; _ . **************** ***************** v.1_ MFRQLRFESTM RDPTAEQFQEELEKFQQAIDSREDPVSCAFVVLMAHGREGFLKGEDGE v .2_ MFRQLRFESTMKRDPTAEQFQEELEKFQQAI DSREDPVSCAFWLMAHGREGFLKGEDGE v .3_ MFRQLRFESTMKRDPTAEQFQEELE FQQAIDSREDPVSCAFWLMAHGREGFLKGE DGE v.4_ ' MFRQLRFESTMKRDPTAEQFQEELEKFQQAI DSREDPVSCAFWLMAHGREGFLKGEDGE v .5_ MFRQLRFESTMKRDPTAEQFQEELEKFQQAI DSREDPVSCAFWLMAHGREGFLKGEDGE v .6_ MFRQLRFESTMKRDPTAEQFQEELEKFQQAI DSREDPVSCAFWLMAHGREGFLKGEDGE ************************************************************ v. l_ MVKLENLFEALNNKNCQALRAKPKVY I IQACRGEQRDPGETVGGDEI VMVI DSPQTIPT v.2_ MVKLENLFEALNNKNCQALRAKPKVY I IQACRGEQRDPGETVGGDEI VMVI DSPQTIPT v.3_ MVKLENLFEALNNKNCQALRAKPKVY I IQACRGATLPSPFPYLSL v.4_ MVKLENLFEALNNKNCQALRAKPKVY II QACRGEQRDPGETVGGDEIVMV I KDSPQT I PT v.5_ ( MVKLENLFEALNNKNCQALRAKPKVY I IQACRGEQRDPGETVGGDEI VMVI DSPQTIPT v.6_ MVKLENLFEAMNNKNCQALRAKPKVYI IQACRGEQRD PGETVGGDEI VMVI DSPQTIPT * * * **** * **·*** * ** * ****** * ****** * * v.1_ YTDALHVYSTVEGY IAYRHDQKGSCFIQTLVDVFTKRKGHILELLTEVTRRMAEAELVQE v .2_ YTDALHVYSTVEGPTPFQDPLYLPSEAPPNPPLWNSQDTSPTDMIRKAH-ALSRPWWMCS v.3_ v .4_ YTDALHVYSTVEGY IAYRHDQKGSCFIQTLVDVFTKRKGHILELLTEVTRRMAEAELVQE v.5_ YTDALHVYSTVEGY IAYRHDQKGSCFIQTLVDVFTKRKGHILELLTEV RRMAEAELVQE v.6_ YTDALHVYSTVEGY I AY RHDQKGSCFIQTLVDVFTKRKGHILELLTEVTRRMAEAELVQE v. l_ GKARKTNPEIQSTLRKRLYLQ v.2_ RRGKDISWNF --v.3_ v.4_ GKARKTNPEIQSTLRKRLYLQ .5_ GKARKTNPEIQSTLRKRLYLQ v.6 GKARKTNPEIQSTLRKRLYLQ Table XXIX. Search Peptides 213P1F11 Variant 1: Nonamers, Decamers, 15-mers (aa.1-242) (SEQ. ID. NO.73) 1 MSNPRSLEEE KYDMSGARLA LILCVTKARE GSEEDLDALE HMFRQLRFES TMKRDPTAEQ 61 FQEELEKFQQ AIDSREDPVS CAFWLMAHG REGFLKGEDG EMVKLENLFE ALNNKNCQAL 121 RAKPKVYIIQ ACRGEQRDPG ETVGGDEIVM VIKDSPQTIP TYTDALHVYS TVEGY IAYRH 181 DQKGSCFIQT LVDVFTKRKG HILELLTEVT RRMAEAELVQ EG ARKTNPE IQSTLRKRLY 241 LQ 213P1F11 Variant 2: Nonamers (aa. 167-230) (SEQ. ID. NO. 74) HVYS TVEGPTPFQD PLYLPSEAPP NPPLWNSQDT SPTDMIRKAH ALSRPWWMCS RRGKDISWNF Decamers (aa. 166-230) (SEQ . ID. NO. 75) LHVYS TVEGPTPFQD PLYLPSEAPP NPPLWNSQDT SPTDMIRKAH ALSRPWWMCS RRGKDISWNF -mers (aa. 161-230) (SEQ. ID. NO. 76) TYTDALHVYS TVEGPTPFQD PLYLPSEAPP NPPLWNSQDT SPTDMIRKAH ALSRPWWMCS RRGKDISWNF 213P1F11 Variant 3: Nonamers (aa. 127-146) (SEQ. ID. NO. 77) YIIQ ACRGATLPSP FPYLSL Decamers (aa.126-146) (SEQ. ID. NO. 78) VYIIQ ACRGATLPSP FPYLSL 15mers (aa. 121-146) (SEQ. ID. NO. 79) RAKPKVYIIQ ACRGATLPSP FPYLSL 213P1F11 Variant 4: Nonamers (aa. 1-94) (SEQ. ID. NO. 80) MGKCQEYDKS LSVQPEKRTG LRDENGECGQ TFRLKEEQGR AFRGSSVHQK LVNDPRETQE VFGGGVGDIV GRDLSISFRN SETSASEEEK YDMS Decamers (aa. 1-95) (SEQ. ID. NO. 81) MGKCQEYDKS LSVQPEKRTG LRDENGECGQ TFRLKEEQGR AFRGSSVHQK LVNDPRETQE VFGGGVGDIV GRDLSISFRN SETSASEEEK YDMSG -mers (aa. 1-100) (SEQ. ID. NO. 82) MGKCQEYDKS LSVQPEKRTG LRDENGECGQ TFRLKEEQGR AFRGSSVHQK LVNDPRETQE VFGGGVGDIV GRDLSISFRN SETSASEEEK YDMS GARLAL 213P1F11 Variant 5: Nonamers (aa. 16-32) (SEQ. ID. NO. 83) GA LA LILRVTKARE GS Decamers (aa. 15-33) (SEQ. ID. NO.. 84) S GAR LA LILRVTKARE GSE -mers (aa. 10-38) ( SEQ . ID. NO.

KYDMSGARLA LILRVTKARE GSEEDLDA 213P1F11 Variant 6: Nonamers (aa. 104-120) (SEQ. ID. NO. 86) KLENLFE AMNNKNCQAL Decamers (aa. 103-121) (SEQ. ID. NO. 87) VKLENLFE AMNNKNCQAL R , -mers (aa. 98-126) (SEQ. ID. NO.

EDG EMVKLENLFE AMNNKNCQAL RAKPKV 164,326/3 327 SEQUENCE LISTING <110> AGENSYS , INC.

CHALLITA-EID, Pia M.

RAITANO, Arthur B.

FARIS, Mary HUBERT, Rene S.

MORRISON, Robert Kendall GE, angmao JAKOBOVITS, Aya <120> NUCLEIC ACID AND CORRESPONDING PROTEIN ENTITLED 213P1F11 USEFUL IN TREATMENT AND DETECTION OF CANCER <130> 511582006640 <140> PCT/US02/10220 <141> 2002-04-01 <160> 88 <170> FastSEQ for Windows Version 4.0 <210> 1 <211> 166 <212> DNA <213> Homo Sapiens <220> <221> misc_feature <222> 156 <223> n = A, T, C or G <400> 1 gatctgaggt tgaggatacc agggaacgct tggaggagta accgggaaga aaagccctga 60 gtttttagtg acgctggatg tgcccgtagg ttacttgagc aggaaaggtc aaagctgggg 120 atggagaggt acaggctgta gagttggaag atagtnaagc tcgatc 166 <210> 2 <211> 3336 <212> DNA <213> Homo Sapiens <220> <221> CDS <222> (404; (1129) <400> 2 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaa atg age aat ccg 415 Met Ser Asn Pro 1 164,326/3 328 egg tct ttg gaa gag gag aaa tat gat atg tea ggt gec cgc ctg gec Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala Arg Leu Ala 10 15 20 eta ata ctg tgt gtc acc aaa gec egg gaa ggt tec gaa gaa gac ctg 511 Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser Glu Glu Asp Leu 30 35 gat get ctg gaa cac atg ttt egg cag ctg aga ttc gaa age acc atg 559 Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu Ser Thr Met 40 45 50 aaa aga gac ccc act gec gag caa ttc cag gaa gag ctg gaa aaa ttc 607 Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu Glu Lys Phe 55 60 65 cag cag gec ate gat tec egg gaa gat ccc gtc agt tgt gec ttc gtg 655 Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser Cys Ala Phe Val 70 75 80 gta etc atg get cac ggg agg gaa ggc ttc etc aag gga gaa gat ggg 703 Val Leu Met Ala His Gly Arg Glu Gly Phe Leu' Lys Gly Glu Asp Gly 85 90 95 100 gag atg gtc aag ctg gag aat etc ttc gag gee ctg aac aac aag aac 751 Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu Asn Asn Lys Asn 105 110 115 tgc cag gee ctg cga get aag ccc aag gtg tac ate ata cag gec tgt 799 Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He He Gin Ala Cys 120 125 130 cga gga gaa caa agg gac ccc ggt gaa aca gta ggt gga gat gag att 847 Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly Gly Asp Glu He 135 140 145 gtg atg gtc ate aaa gac age cca caa acc ate cca aca tac aca gat 895 Val Met Val He Lys Asp Ser Pro Gin Thr He Pro Thr Tyr Thr Asp 150 155 160 gec ttg cac gtt tat tec acg gta gag gga tac ate gec tac cga cat 943 Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He Ala Tyr Arg His 165 170 175 180 gat cag aaa ggc tea tgc ttt ate cag acc ctg gtg gat gtg ttc acg 991 Asp Gin Lys Gly Ser Cys: Phe He Gin Thr Leu Val Asp Val Phe Thr 185 190 195 aag agg aaa gga cat ate ttg gaa ctt ctg aca gag gtg acc egg egg 1039 Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu Val Thr Arg Arg 200 205 210 atg gca gaa gca gag ctg gtt caa gaa gga aaa gca agg aaa acg aac 1087 Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala Arg Lys Thr Asn 215 220 225 cct gaa ate caa age acc etc egg aaa egg ctg tat ctg cag 1129 Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr Leu Gin 230 235 240 tagaagtaga aagaccagga ggagctttcc ttccagcatt ctttctgtct cacagaaatt 1189 164,326/3 329 tagaggcagc tcttacctct ccccaagatc ttctgttccc aaggccaaat ggcacccagt 1249 ttcttttcca tcacaccctt catgcaggtc ctcctgtcct tattagagca agccagccaa 1309 aacttagcac aaggcatggt ggcaacatta acatcacctc cctcaggctg gactttctat 1369 ctttattaat gcaaccgaag agacctaaga gtgcattcac ttatcccact ttctgttcct 1429 gtggtcttct ttctcccatg aagcagaaac tggataaagc tcaagatttt ccatagacaa 1489 accaaagccc actcatcccc tcctacccca atccaacctc tgctggctcc tgcatctcac 1549 ttggaggtca aacctcctcc tgaggccaat gcattcccaa cttccagttc tttcctttac 1609 cctggagagt tagtaaggta agaaccattc tttctctcca aaaccactcc tccttggctg 1669 gcaagttggt gtcctaactc cgttctcttc ctagctcatg gcctctctag ataataaagt 1729 tgtctcctcc tttctggatc tcttcctcct aacacccctc ccctgaaacc ctggactctg 1789 ccctctctcc aagaaaatcc atctattcaa ctattcttgc attcaattac tctaaatgag 1849 agcgtgttgg agctatggca aattccctgt tgtcaccttg ctattttgca gacaacataa 1909 tatttaacct ctcataacca gagaggttaa ataatttgtc aaatgcaata cagtaagaca 1969 gaggcaagga caaggtttga cttccagccc agcctctttt ccacaacctg ctaaatcctg 2029 atccatctga aaacttttct aattagtgaa gatgactaat aaaaattttc cctatctcca 2089 aggtaggagc tttctggaag tttctagaaa ttttcaataa ccaccagcca aggttacctc 2149 caggtaacct tgcagcacca ggctggaagt cagatcggct tcactatctt ccaactctac 2209 agcctgtatc tctccatccc cagctttgac ctttcctgct caagtaacct acgggcacat 2269 ccagcgtcac taaaaactca gggcttttct tcccggttac tcctccaagc gttccctggt 2329 atcctcaacc tcagatccca ggttcagatt tctgcagtca atctatgacc cctctcttct 2389 tgcatcctt.c atatgccacc agacaccatg cccagtccag cctgattttg aaacaacttt 2449 catgccggtc ttctcttccc tgacatgtta ctgtccaggc tcaagtcctc agcttctcat 2509 atctgcatct ttgcaaccaa cttcctccct tgcctctctg cttttccatc ccacttttca 2569 tgtgtcctcc ataccatcta taacagtgat ctccctggaa cactcaagaa gacacaacat 2629 accatattat ttaaagacca gggtactgga cagtggctca cacctgtatt cccgactttg 2689 agagtctgaa gcgggaggat cacttgaggc caggagttaa gagaccagcc tgggcaacac 2749 agcaagaccc tgtctctaaa aaaaaaaatt aattaactgg gtatggtggc acatgcctgt 2809 agtcccagct actcaggagg ctgaggtggg aggatgactt gagcccagga gtttgaggct 2869 gcaaggagct atgatcatgc cagtgcatcc cagctctagg tgagacagtg agatccggtc 2929 tccaaaataa atcaatcaat caaataaaga ccaaagtcaa accgcacatc aggatctctc 2989 acacccttcc aattttgcca tctaccagca cttagctaaa cccatctccc atctcttcca 3049 ccatgaattc actctttcaa aaaggctaat gtcttcttac tcacccttgc ctctaagcct 3109 ttgctatcac catttccccc aagctggagg gccctccctc tccctttacc cctcttccac 3169 tacctcccac ccctactttt tccagaaagc catttcctct cttttttctg attgatcctt 3229 ccctctcacc caggattaga tgctggaaat gaccacttct ggagggcagg gaacaagccc 3289 ttaatctgca taatgagtgt tcaataaaca gttgtcaaac tttgaaa 3336 <210> 3 <211> 242 <212> PRT <213> Homo Sapiens <400> 3 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 164,326/3 330 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 4 <211> 3410 <212> DNA <213> Homo Sapiens <220> <221> CDS <222> (404) . (1093) <400> 4 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gecaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaa atg age aat ccg 415 Met Ser Asn Pro 1 egg tct ttg gaa gag gag aaa tat gat atg tea ggt gec cgc ctg gec 463 Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala Arg Leu Ala 10 15 20 eta ata ctg tgt gtc acc aaa gee egg gaa ggt tec gaa gaa gac ctg 511 Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser Glu Glu Asp Leu 30 35 gat get ctg gaa cac atg ttt egg cag ctg aga ttc gaa age acc atg 559 Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu Ser Thr Met 40 45 50 aaa aga gac ccc act gee gag caa ttc cag gaa gag ctg gaa aaa ttc 607 Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu Glu Lys Phe 55 60 65 cag cag gee ate gat tec egg gaa gat ccc gtc agt tgt gec ttc gtg 655 Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser Cys Ala Phe Val 70 75 80 gta etc atg get cac ggg agg gaa ggc ttc etc aag gga gaa gat ggg 703 Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys Gly Glu Asp Gly 85 90 95 100 gag atg gtc aag ctg gag aat etc ttc gag gee ctg aac aac aag aac 751 164,326/3 331 Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu Asn Asn Lys Asn 105 110 115 tgc cag gcc ctg cga get aag ccc aag gtg tac ate ata cag gec tgt 799 Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr lie lie Gin Ala Cys 120 125 130 cga gga gaa caa agg gac ccc ggt gaa aca gta ggt gga gat gag att 847 Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly Gly Asp Glu He 135 140 145 gtg atg gtc ate aaa gac age cca caa acc ate cca aca tac aca gat 895 Val Met Val He Lys Asp Ser Pro Gin Thr He Pro Thr Tyr Thr Asp 150 155 160 gcc ttg cac gtt tat tec acg gta gag gga ccc acg ccc ttc cag gat 943 Ala Leu His Val Tyr Ser Thr Val Glu Gly Pro Thr Pro Phe Gin Asp 165 170 175 180 ccc etc tac eta ccc tct gaa get ccc ccg aac cca cct etc tgg aat 991 Pro Leu Tyr Leu Pro Ser Glu Ala Pro Pro Asn Pro Pro Leu Trp Asn 185 190 195 tec cag gat aca teg cct acc gac atg ate aga aag get cat get tta 1039 Ser Gin Asp Thr Ser Pro Thr Asp Met He Arg Lys Ala His Ala Leu 200 205 210 tec aga ccc tgg tgg atg tgt tea cga aga gga aag gac ata tct tgg 1087 Ser Arg Pro Trp Trp Met Cys Ser Arg Arg Gly Lys Asp He Ser Trp 215 220 225 aac ttc tgacagaggt gacccggcgg atggcagaag cagagctggt tcaagaagga 1143 Asn Phe 230 aaagcaagga aaacgaaccc tgaaatccaa agcaccctcc ggaaacggct gtatctgcag 1203 tagaagtaga aagaccagga ggagctttcc ttccagcatt ctttctgtct cacagaaatt 1263 tagaggcagc tcttacctct ccccaagatc ttctgttccc aaggccaaat ggcacccagt 1323 ttcttttcca tcacaccctt catgcaggtc ctcctgtcct tattagagca agccagccaa 1383 aacttagcac aaggcatggt ggcaacatta acatcacctc cctcaggctg gactttctat 1443 ctttattaat gcaaccgaag agacctaaga gtgeattcac ttatcccact ttctgttcct 1503 gtggtcttct ttctcccatg aagcagaaac tggataaagc tcaagatttt ccatagacaa 1563 accaaagccc actcatcccc tcctacccca atccaacctc tgctggctcc tgcatctcac 1623 ttggaggtca aacctcctcc tgaggccaat gcattcccaa cttccagttc tttcctttac 1683 cctggagagt tagtaaggta agaaccattc tttctctcca aaaccactcc tccttggctg 1743 gcaagttggt gtcctaactc cgttctcttc ctagctcatg gcctctctag ataataaagt 1803 tgtctcctcc tttctggatc tcttcctcct aacacccctc ccctgaaacc ctggactctg 1863 ccctctctcc aagaaaatcc atctattcaa etattcttge attcaattac tctaaatgag 1923 agcgtgttgg agctatggca aattccctgt tgtcaccttg etattttgea gacaacataa 1983 tatttaacct ctcataacca gagaggttaa ataatttgtc aaatgeaata cagtaagaca 2043 gaggcaagga -caaggtttga cttccagccc agectctttt ccacaacctg ctaaatcctg 2103 atccatctga aaacttttct aattagtgaa gatgactaat aaaaattttc cctatctcca 2163 aggtaggagc tttctggaag tttctagaaa ttttcaataa ccaccagcca aggttacctc 2223 caggtaacct tgcagcacca ggctggaagt cagategget tcactatctt ccaactctac 2283 agectgtate tctccatccc cagctttgac ctttcctgct caagtaacct aegggcacat 2343 ccagcgtcac taaaaactca gggcttttct tcccggttac tcctccaagc gttccctggt 2403 atcctcaacc tcagatccca ggttcagatt tetgeagtea atctatgacc cctctcttct 2463 tgcatccttc atatgccacc agacaccatg cccagtccag cctgattttg aaacaacttt 2523 catgccggtc ttctcttccc tgacatgtta ctgtccaggc tcaagtcctc agcttctcat 2583 atetgeatet ttgcaaccaa cttcctccct tgcctctctg cttttccatc ccacttttca 2643 tgtgtcctcc ataccatcta taacagtgat ctccctggaa cactcaagaa gacacaacat 2703 164,326/3 332 accatattat ttaaagacca gggtactgga cagtggctca cacctgtatt cccgactttg 2763 agagtctgaa gcgggaggat cacttgaggc caggagttaa gagaccagcc tgggcaacac 2823 agcaagaccc tgtctctaaa aaaaaaaatt aattaactgg gtatggtggc acatgcctgt 2883 agtcccagct actcaggagg ctgaggtggg aggatgactt gagcccagga gtttgaggct 2943 gcaaggagct atgatcatgc cagtgcatcc cagctctagg tgagacagtg agatccggtc 3003 tccaaaataa atcaatcaat caaataaaga ccaaagtcaa accgcacatc aggatctctc 3063 acacccttcc aattttgcca tctaccagca cttagctaaa cccatctccc atctcttcca 3123 ccatgaattc actctttcaa aaaggctaat gtcttcttac tcacccttgc ctctaagcct 3183 ttgctatcac catttccccc aagctggagg gccctccctc tccctttacc cctcttccac 3243' tacctcccac ccctactttt tccagaaagc catttcctct cttttttctg attgatcctt 3303 ccctctcacc caggattaga tgctggaaat gaccacttct ggagggcagg gaacaagccc 3363 ttaatctgca taatgagtgt tcaataaaca gttgtcaaac tttgaaa 3410 <210> 5 <211> 230 <212> PRT <213> Homo Sapiens <400> 5 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Pro Thr 165 170 175 Pro Phe Gin Asp Pro Leu Tyr Leu Pro Ser Glu Ala Pro Pro Asn Pro 180 185 190 Pro Leu Trp Asn Ser Gin Asp Thr Ser Pro Thr Asp Met lie Arg Lys 195 200 205 Ala His Ala Leu Ser Arg Pro Trp Trp Met Cys Ser Arg Arg Gly Lys 210 215 220 Asp He Ser Trp Asn Phe 225 230 <210> 6 <211> 3404 <212> DNA <213> Homo Sapiens <220> <221> CDS <222> (404) ... (841) <400> 6 164,326/1 333 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gecaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaa atg age aat ccg 415 Met Ser Asn Pro 1 egg tct ttg gaa gag gag aaa tat gat atg tea ggt gee cgc ctg gee 463 Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala Arg Leu Ala 10 15 20 eta ata ctg tgt gtc ace aaa gee egg gaa ggt tec gaa gaa gac ctg 511 Leu lie Leu Cys Val Thr Lys Ala Arg Glu Gly Ser Glu Glu Asp Leu 30 35 gat get ctg gaa cac atg ttt egg cag ctg aga ttc gaa age ace atg 559 Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu Ser Thr Met 40 45 50 aaa aga gac ccc act gee gag caa ttc cag gaa gag ctg gaa aaa ttc 607 Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu Glu Lys Phe 55 60 65 cag cag gee ate gat tec egg gaa gat ccc gtc agt tgt gec ttc gtg 655 Gin Gin Ala lie Asp Ser Arg Glu Asp Pro Val Ser Cys Ala Phe Val 70 75 80 gta etc atg get cac ggg agg gaa ggc ttc etc aag gga gaa gat ggg 703 Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys Gly Glu Asp Gly 85 90 95 100 gag atg gtc aag ctg gag aat etc ttc gag gee ctg aac aac aag aac 751 Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu Asn Asn Lys Asn 105 110 115 tgc cag gec ctg cga get aag ccc aag gtg tac ate ata cag gee tgt 799 Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He He Gin Ala Cys 120 125 130 cga gga gee acc ctg ccc age ccc ttt cct tac ctt tct etc 841 Arg Gly Ala Thr Leu Pro Ser Pro Phe Pro Tyr Leu Ser Leu 135 140 145 tgactttgee tcctcctctt cttgttgttt cagaacaaag ggaccccggt gaaacagtag 901 gtggagatga gattgtgatg gtcatcaaag acagcccaca aaccatccca acatacacag 961 atgccttgca cgtttattcc acggtagagg gatacatcgc ctaccgacat gatcagaaag 1021 gctcatgctt tatccagacc ctggtggatg tgttcacgaa gaggaaagga catatcttgg 1081 aacttctgac agaggtgacc eggeggatgg cagaagcaga gctggttcaa gaaggaaaag 1141 caaggaaaac gaaccctgaa atccaaagca ccctccggaa acggctgtat ctgcagtaga 1201 agtagaaaga ccaggaggag ctttccttcc agcattcttt ctgtctcaca gaaatttaga 1261 ggcagctctt acctctcccc aagatcttct gttcccaagg ccaaatggca cccagtttct 1321 tttccatcac acccttcatg caggtcctcc tgtccttatt agagcaagee agecaaaact 1381 tagcacaagg catggtggca acattaacat cacctccctc aggctggact ttctatcttt 1441 attaatgeaa ccgaagagac ctaagagtgc attcacttat cccactttct gttcctgtgg 1501 tcttctttct cccatgaagc agaaactgga taaagctcaa gattttccat agacaaacca 1561 aagcccactc atcccctcct accccaatcc aacctctgct ggctcctgca tctcacttgg 1621 aggtcaaacc tcctcctgag gecaatgeat tcccaacttc cagttctttc ctttaccctg 1681 gagagttagt aaggtaagaa ccattctttc tctccaaaac cactcctcct tggctggcaa 1741 164,326/1 334 gttggtgtcc taactccgtt ctcttcctag ctcatggcct ctctagataa taaagttgtc 1801 tcctcctttc tggatctctt cctcctaaca cccctcccct gaaaccctgg actctgccct 1861 ctctccaaga aaatccatct attcaactat tcttgcattc aattactcta aatgagagcg 1921 tgttggagct atggcaaatt ccctgttgtc accttgctat tttgcagaca acataatatt 1981 taacctctca taaccagaga ggttaaataa tttgtcaaat gcaatacagt aagacagagg 2041 caaggacaag gtttgacttc cagcccagcc tcttttccac aacctgctaa atcctgatcc 2101 atctgaaaac ttttctaatt agtgaagatg actaataaaa attttcccta tctccaaggt 2161 aggagctttc tggaagtttc tagaaatttt caataaccac cagccaaggt tacctccagg 2221 taaccttgca gcaccaggct ggaagtcaga tcggcttcac tatcttccaa ctctacagcc 2281 tgtatctctc catccccagc tttgaccttt cctgctcaag taacctacgg gcacatccag 2341 cgtcactaaa aactcagggc ttttcttccc ggttactcct ccaagcgttc cctggtatcc 2401 tcaacctcag atcccaggtt cagatttctg cagtcaatct atgacccctc tcttcttgca 2461 tccttcatat gccaccagac accatgccca gtccagcctg attttgaaac aactttcatg 2521 ccggtcttct cttccctgac atgttactgt ccaggctcaa gtcctcagct tctcatatct 2581 gcatctttgc aaccaacttc ctcccttgcc tctctgcttt tccatcccac ttttcatgtg 2641 tcctccatac catctataac agtgatctcc ctggaacact caagaagaca caacatacca 2701 tattatttaa agaccagggt actggacagt ggctcacacc tgtattcccg actttgagag 2761 tctgaagcgg gaggatcact tgaggccagg agttaagaga ccagcctggg caacacagca 2821 agaccctgtc tctaaaaaaa aaaattaatt aactgggtat ggtggcacat gcctgtagtc 2881 ccagctactc aggaggctga ggtgggagga tgacttgagc ccaggagttt gaggctgcaa 2941 ggagctatga tcatgccagt gcatcccagc tctaggtgag acagtgagat ccggtctcca 3001 aaataaatca atcaatcaaa taaagaccaa agtcaaaccg cacatcagga tctctcacac 3061 ccttccaatt ttgccatcta ccagcactta gctaaaccca tctcccatct cttccaccat 3121 gaattcactc tttcaaaaag gctaatgtct tcttactcac ccttgcctct aagcctttgc 3181 tatcaccatt tcccccaagc tggagggccc tccctctccc tttacccctc ttccactacc 3241 tcccacccct actttttcca gaaagccatt tcctctcttt tttctgattg atccttccct 3301 ctcacccagg attagatgct ggaaatgacc acttctggag ggcagggaac aagcccttaa 3361 tctgcataat gagtgttcaa taaacagttg tcaaactttg aaa 3404 <210> 7 <211> 146 <212> PRT <213> Homo Sapiens <400> 7 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Ala Thr Leu Pro Ser Pro Phe Pro Tyr Leu 130 135 140 Ser Leu 145 <210> 8 <211> 966 <212> DNA <213> Homo Sapiens 164,326/1 335 <220> <221> CDS <222> (1) ... (963) <400> 8 atg ggg aaa tgc caa gag tat gac aaa agt ctg tct gtg cag cca gag 48 Met Gly Lys Cys Gin Glu Tyr Asp Lys Ser Leu Ser Val Gin Pro Glu 1 5 10 15 aag aga aca gga etc aga gat gag aat gga gaa tgt gga cag aca ttc 96 Lys Arg Thr Gly Leu Arg Asp Glu Asn Gly Glu Cys Gly Gin Thr Phe 25 30 aga etc aag gaa gag caa ggg agg get ttc agg gga agt tea gtc cac 144 Arg Leu Lys Glu Glu Gin Gly Arg Ala Phe Arg Gly Ser Ser Val His 40 45 cag aag ctg gtg aat gac cca egg gag aca cag gaa gtt ttt. ggg ggc 192 Gin Lys Leu Val Asn Asp Pro Arg Glu Thr Gin Glu Val Phe Gly Gly 50 55 60 gga gtg ggg gac att gtg gga egg gat etc agt att age ttc aga aac 240 Gly Val Gly Asp lie Val Gly Arg Asp Leu Ser He Ser Phe Arg Asn 65 70 75 80 tct gag acc tct gca agt gag gag gag aaa tat gat atg tea ggt gcc 288 Ser Glu Thr Ser Ala Ser Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala 85 90 95 cgc ctg gcc eta ata ctg tgt gtc acc aaa gcc egg gaa ggt tec gaa 336 Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser Glu 100 105 110 gaa gac ctg gat get ctg gaa cac atg ttt egg cag ctg aga ttc gaa 384 Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu 115 120 125 age acc atg aaa aga gac ccc act gcc gag caa ttc cag gaa gag ctg 432 Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu 130 135 140 gaa aaa ttc cag cag gcc ate gat tec egg gaa gat ccc gtc agt tgt 480 Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser Cys 145 150 155 160 gcc ttc gtg gta etc atg get cac ggg agg gaa ggc ttc etc aag gga 528 Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys Gly 165 170 175 gaa gat ggg gag atg gtc aag ctg gag aat etc ttc gag gcc ctg aac 576 Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu Asn 180 185 190 aac aag aac tgc cag gcc ctg cga get aag ccc aag gtg tac ate ata 624 Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He He 195 200 205 cag gcc tgt cga gga gaa caa agg gac ccc ggt gaa aca gta ggt gga 672 Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly Gly 210 215 220 164,326/1 336 gat gag att gtg atg gtc ate aaa gac age cca caa acc ate cca aca 720 Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro Thr 225 230 235 240 tac aca gat gee ttg cac gtt tat tec acg gta gag gga tac ate gec 768 Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He Ala 245 250 255 tac cga cat gat cag aaa ggc tea tgc ttt ate cag acc ctg gtg gat 816 Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val Asp 260 265 270 gtg ttc acg aag agg aaa gga cat ate ttg gaa ctt ctg aca gag gtg 864 Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu Val 275 280 285 acc egg egg atg gca gaa gca gag ctg gtt caa gaa gga aaa gca agg 912 Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala Arg 290 295 300 aaa acg aac cct gaa ate caa age acc etc egg aaa egg ctg tat ctg 960 Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr Leu 305 310 315 320 cag tag 966 Gin <210> 9 <211> 321 <212> PRT <213> Homo Sapiens <400> 9 Met Gly Lys Cys Gin Glu Tyr Asp Lys Ser Leu Ser Val Gin Pro Glu 1 5 10 15 Lys Arg Thr Gly Leu Arg Asp Glu Asn Gly Glu Cys Gly Gin Thr Phe 25 30 Arg Leu Lys Glu Glu Gin Gly Arg Ala Phe Arg Gly Ser Ser Val His 40 45 Gin Lys Leu Val Asn Asp Pro Arg Glu Thr Gin Glu Val Phe Gly Gly 50 55 60 Gly Val Gly Asp He Val Gly Arg Asp Leu Ser He Ser Phe Arg Asn 65 70 75 80 Ser Glu Thr Ser Ala Ser Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala 85 90 95 Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser Glu 100 105 110 Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu 115 120 125 Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu 130 135 140 Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser Cys 145 150 155 160 Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys Gly 165 170 175 Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu Asn 180 185 190 Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He He 195 200 205 164,326/1 337 Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly Gly 210 215 220 Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro Thr 225 230 235 240 Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He Ala 245 250 255 Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val Asp 260 265 270 Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu Val 275 280 285 Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala Arg 290 295 300 Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr Leu 305 310 315 320 Gin <210> 10 <211> 3336 <212> DNA <213> Homo Sapiens <220> <221> CDS <222> (404) ... (1129) <400> 10 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaa atg age aat ccg 415 Met Ser Asn Pro 1 egg tct ttg gaa gag gag aaa tat gat atg tea ggt gec cgc ctg gec 463 Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala Arg Leu Ala 10 15 20 eta ata ctg cgt gtc ace aaa gec egg gaa ggt tec gaa gaa gac ctg 511 Leu He Leu Arg Val Thr Lys Ala Arg Glu Gly Ser Glu Glu Asp Leu 30 35 gat get ctg gaa cac atg ttt egg cag ctg aga ttc gaa age acc atg 559 Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu Ser Thr Met 40 45 50 aaa aga gac ccc act gec gag caa ttc cag gaa gag ctg gaa aaa ttc 607 Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu Glu Lys Phe 55 60 65 cag cag gec ate gat tec egg gaa gat ccc gtc agt tgt gee ttc gtg 655 Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser Cys Ala Phe Val 70 75 80 gta etc atg get cac ggg agg gaa ggc ttc etc aag gga gaa gat ggg 703 Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys Gly Glu Asp Gly 85 90 95 100 164,326/1 338 gag atg gtc aag etg gag aat etc ttc gag gee etg aac aac aag aac 751 Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu Asn Asn Lys Asn 105 110 115 tgc eag gee etg cga get aag ccc aag gtg tac ate ata eag gee tgt 799 Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr lie lie Gin Ala Cys 120 125 130 cga gga gaa caa agg gac ccc ggt gaa aca gta ggt gga gat gag att 847 Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly Gly Asp Glu lie 135 140 145 gtg atg gtc ate aaa gac age cca caa ace ate cca aca tac aca gat 895 Val Met Val lie Lys Asp Ser Pro Gin Thr lie Pro Thr Tyr Thr Asp 150 155 160 gec ttg cac gtt tat tec acg gta gag gga tac ate gee tac cga cat 943 Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr lie Ala Tyr Arg His 165 170 175 180 gat cag aaa ggc tea tgc ttt ate cag ace etg gtg gat gtg ttc acg 991 Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val Asp Val Phe Thr 185 190 195 aag agg aaa gga cat ate ttg gaa ctt etg aca gag gtg acc egg egg 1039 Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu Val Thr Arg Arg 200 205 210 atg gca gaa gca gag etg gtt caa gaa gga aaa gca agg aaa acg aac 1087 Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala Arg Lys Thr Asn 215 220 225 cct gaa ate caa age acc etc egg aaa egg etg tat etg cag 1129 Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr Leu Gin 230 235 240 tagaagtaga aagaccagga ggagctttcc ttccagcatt ctttctgtct cacagaaatt 1189 tagaggcagc tcttacctct ccccaagatc ttctgttccc aaggccaaat ggcacccagt 1249 ttcttttcca tcacaccctt catgeaggtc ctcctgtcct tattagagca agccagccaa 1309 aacttagcac aaggcatggt ggcaacatta acatcacctc cctcaggctg gactttctat 1369 ctttattaat gcaaccgaag agacctaaga gtgeattcac ttatcccact ttctgttcct 1429 gtggtcttct ttctcccatg aagcagaaac tggataaagc tcaagatttt ccatagacaa 1489 accaaagccc actcatcccc tcctacccca atccaacctc tgctggctcc tgcatctcac 1549 ttggaggtca aacctcctcc tgaggccaat gcattcccaa cttccagttc tttcctttac 1609 cctggagagt tagtaaggta agaaccattc tttctctcca aaaccactcc tccttggctg 1669 gcaagttggt gtcctaactc cgttctcttc ctagctcatg gcctctctag ataataaagt 1729 tgtctcctcc tttctggatc tcttcctcct aacacccctc ccctgaaacc ctggactctg 1789 ccctctctcc aagaaaatcc atctattcaa etattcttge attcaattac tctaaatgag 1849 agcgtgttgg agctatggca aattccctgt tgtcaccttg etattttgea gacaacataa 1909 tatttaacct ctcataacca gagaggttaa ataatttgtc aaatgeaata cagtaagaca 1969 gaggcaagga caaggtttga cttccagccc agectctttt ccacaacctg ctaaatcctg 2029 atccatctga aaacttttct aattagtgaa gatgactaat aaaaattttc cctatctcca 2089 aggtaggagc tttctggaag tttctagaaa ttttcaataa ccaccagcca aggttacctc 2149 caggtaacct tgcagcacca ggctggaagt cagategget tcactatctt ccaactctac 2209 agectgtate tctccatccc cagctttgac ctttcctgct caagtaacct aegggcacat 2269 ccagcgtcac taaaaactca gggcttttct tcccggttac tcctccaagc gttccctggt 2329 atcctcaacc tcagatccca ggttcagatt tetgeagtea atctatgacc cctctcttct 2389 tgcatccttc atatgccacc agacaccatg cccagtccag cctgattttg aaacaacttt 2449 catgccggtc ttctcttccc tgacatgtta ctgtccaggc tcaagtcctc agcttctcat 2509 atetgeatet ttgcaaccaa cttcctccct tgcctctctg cttttccatc ccacttttca 2569 tgtgtcctcc ataccatcta taacagtgat ctccctggaa cactcaagaa gacacaacat 2629 164,326/1 339 accatattat ttaaagacca gggtactgga cagtggctca cacctgtatt cccgactttg 2689 agagtctgaa gcgggaggat cacttgaggc caggagttaa gagaccagcc tgggcaacac 2749 agcaagaccc tgtctctaaa aaaaaaaatt aattaactgg gtatggtggc acatgcctgt 2809 agtcccagct actcaggagg ctgaggtggg aggatgactt gagcccagga gtttgaggct 2869 gcaaggagct atgatcatgc cagtgcatcc cagctctagg tgagacagtg agatccggtc 2929 tccaaaataa atcaatcaat caaataaaga ccaaagtcaa accgcacatc aggatctctc 2989 acacccttcc aattttgcca tctaccagca cttagctaaa cccatctccc atctcttcca 3049 ccatgaattc actctttcaa aaaggctaat gtcttcttac tcacccttgc ctctaagcct 3109 ttgctatcac catttccccc aagctggagg gccctccctc tccctttacc cctcttccac 3169 tacctcccac ccctactttt tccagaaagc catttcctct cttttttctg attgatcctt 3229 ccctctcacc caggattaga tgctggaaat gaccacttct ggagggcagg gaacaagccc 3289 ttaatctgca taatgagtgt tcaataaaca gttgtcaaac tttgaaa 3336 <210> 11 <211> 242 <212> PRT <213> Homo Sapiens <400> 11 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Arg Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 12 <211> 3336 <212> DNA <213> Homo Sapiens <220> <221> CDS <222> (404) ... (1129) 164,326/1 340 <400> 12 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaa atg age aat ccg 415 Met Ser Asn Pro 1 egg tct ttg gaa gag gag aaa tat gat atg tea ggt gec cgc ctg gee 463 Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala Arg Leu Ala 10 15 20 eta ata ctg tgt gtc acc aaa gee egg gaa ggt tec gaa gaa gac ctg 511 Leu lie Leu Cys Val Thr Lys Ala Arg Glu Gly Ser Glu Glu Asp Leu 30 35 gat get ctg gaa cac atg ttt egg cag ctg aga ttc gaa age acc atg 559 Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu Ser Thr Met 40 45 50 aaa aga gac ccc act gee gag caa ttc cag gaa gag ctg gaa aaa ttc 607 Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu Glu Lys Phe 55 60 65 cag cag gee ate gat tec egg gaa gat ccc gtc agt tgt gee ttc gtg 655 Gin Gin Ala lie Asp Ser Arg Glu Asp Pro Val Ser Cys Ala Phe Val 70 75 80 gta etc atg get cac ggg agg gaa ggc ttc etc aag gga gaa gat ggg 703 Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys Gly Glu Asp Gly 85 90 95 100 gag atg gtc aag ctg gag aat etc ttc gag gee atg aac aac aag aac 751 Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Met Asn Asn Lys Asn 105 110 115 tgc cag gee ctg cga get aag ccc aag gtg tac ate ata cag gec tgt 799 Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr lie lie Gin Ala Cys 120 125 130 cga gga gaa caa agg gac ccc ggt gaa aca gta ggt gga gat gag att 847 Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly Gly Asp Glu He 135 140 145 gtg atg gtc ate aaa gac age cca caa acc ate cca aca tac aca gat 895 Val Met Val He Lys Asp Ser Pro Gin Thr He Pro Thr Tyr Thr Asp 150 155 160 gec ttg cac gtt tat tec acg gta gag gga tac ate gee tac cga cat 943 Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He Ala Tyr Arg His 165 170 175 180 gat cag aaa ggc tea tgc ttt ate cag acc ctg gtg gat gtg ttc acg 991 Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val Asp Val Phe Thr 185 190 195 aag agg aaa gga cat ate ttg gaa ctt ctg aca gag gtg acc egg egg 1039 Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu Val Thr Arg Arg 164,326/1 341 200 205 210 atg gca gaa gca gag ctg gtt caa gaa gga aaa gca agg aaa acg aac 1087 Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala Arg Lys Thr Asn 215 220 225 cct gaa ate caa age acc etc egg aaa egg ctg tat ctg cag 1129 Pro Glu lie Gin Ser Thr Leu Arg Lys Arg Leu Tyr Leu Gin 230 235 240 tagaagtaga aagaccagga ggagctttcc ttccagcatt ctttctgtct cacagaaatt 1189 tagaggcagc tcttacctct ccccaagatc ttctgttccc aaggccaaat ggcacccagt 1249 ttcttttcca tcacaccctt catgcaggtc ctcctgtcct tattagagca agccagccaa 1309 aacttagcac aaggcatggt ggcaacatta acatcacctc cctcaggctg gactttctat 1369 ctttattaat gcaaccgaag agacctaaga gtgeattcac ttatcccact ttctgttcct 1429 gtggtcttct ttctcccatg aagcagaaac tggataaagc tcaagatttt ccatagacaa 1489 accaaagccc actcatcccc tcctacccca atccaacctc tgctggctcc tgcatctcac 1549 ttggaggtca aacctcctcc tgaggccaat gcattcccaa cttccagttc tttcctttac 1609 cctggagagt tagtaaggta agaaccattc tttctctcca aaaccactcc tccttggctg 1669 gcaagttggt gtcctaactc cgttctcttc ctagctcatg gcctctctag ataataaagt 1729 tgtctcctcc tttctggatc tcttcctcct aacacccctc ccctgaaacc ctggactctg 1789 ccctctctcc aagaaaatcc atctattcaa etattcttge attcaattac tctaaatgag 1849 agcgtgttgg agctatggca aattccctgt tgtcaccttg etattttgea gacaacataa 1909 tatttaacct ctcataacca gagaggttaa ataatttgtc aaatgeaata cagtaagaca 1969 gaggcaagga caaggtttga cttccagccc agectctttt ccacaacctg ctaaatcctg 2029 atccatctga aaacttttct aattagtgaa gatgactaat aaaaattttc cctatctcca 2089 aggtaggagc tttctggaag tttctagaaa ttttcaataa ccaccagcca aggttacctc 2149 caggtaacct tgcagcacca ggctggaagt cagategget tcactatctt ccaactctac 2209 agectgtate tctccatccc cagctttgac ctttcctgct caagtaacct aegggcacat 2269 ccagcgtcac taaaaactca gggcttttct tcccggttac tcctccaagc gttccctggt 2329 atcctcaacc tcagatccca ggttcagatt tetgeagtea atctatgacc cctctcttct 2389 tgcatccttc atatgccacc agacaccatg cccagtccag cctgattttg aaacaacttt 2449 catgccggtc ttctcttccc tgacatgtta ctgtccaggc tcaagtcctc agcttctcat 2509 atetgeatet ttgcaaccaa cttcctccct tgcctctctg cttttccatc ccacttttca 2569 tgtgtcctcc ataccatcta taacagtgat ctccctggaa cactcaagaa gacacaacat 2629 accatattat ttaaagacca gggtactgga cagtggctca cacctgtatt cccgactttg 2689 agagtctgaa gegggaggat cacttgaggc caggagttaa gagaccagcc tgggcaacac 2749 agcaagaccc tgtctctaaa aaaaaaaatt aattaactgg gtatggtggc acatgectgt 2809 agtcccagct actcaggagg ctgaggtggg aggatgactt gageccagga gtttgaggct 2869 gcaaggagct atgatcatgc cagtgcatcc cagctctagg tgagacagtg agatceggtc 2929 tccaaaataa atcaatcaat caaataaaga ccaaagtcaa accgcacatc aggatctctc 2989 acacccttcc aattttgeca tctaccagca cttagctaaa cccatctccc atctcttcca 3049 ccatgaattc actctttcaa aaaggctaat gtcttcttac tcacccttgc ctctaagcct 3109 ttgetatcac catttccccc aagctggagg gccctccctc tccctttacc cctcttccac 3169 tacctcccac ccctactttt tccagaaagc catttcctct cttttttctg attgatcctt 3229 ccctctcacc caggattaga tgctggaaat gaccacttct ggagggcagg gaacaagccc 3289 ttaatctgea taatgagtgt tcaataaaca gttgtcaaac tttgaaa 3336 <210> 13 <211> 242 <212> PRT <213> Homo Sapiens <400> 13 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu lie Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala- Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 164,326/1 342 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Met 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235- 240 Leu Gin <210> 14 <211> 3336 <212> DNA <213> Homo Sapiens <220> <221> CDS <222> (404) (1129) <400> 14 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaa atg agc aat ccg 415 Met Ser Asn Pro 1 egg tct ttg gaa gag gag aaa tat gat atg tea ggt gec cgc ctg gec 463 Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala Arg Leu Ala 10 15 20 eta ata ctg tgt gtc acc aaa gec egg gaa ggt tec gaa gaa gac ctg 511 Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser Glu Glu Asp Leu 30 35 gat get ctg gaa cac atg ttt egg cag ctg aga ttc gaa age acc atg 559 Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu Ser Thr Met 40 45 50 aaa aga gac ccc act gec gag caa ttc cag gaa gag ctg gaa aaa ttc 607 Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu Glu Lys Phe 55 60 65 cag cag gcc ate gat tec egg gaa gat ccc gtc agt tgt gee ttc gtg 655 Gin Gin Ala lie Asp Ser Arg Glu Asp. Pro Val Ser Cys Ala Phe Val 70 75 80 gta etc atg get cac ggg agg gaa ggc ttc etc aag gga gaa gat ggg 703 Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys Gly Glu Asp Gly 85 90 95 100 gag atg gtc aag ctg gag aat etc ttc gag gcc ctg aac aac aag aac 751 Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu Asn Asn Lys Asn 105 110 115 tgc cag gcc ctg cga get aag ccc aag gtg tac ate ata cag gcc tgt 799 Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr lie lie Gin Ala Cys 120 125 130 cga gga gaa caa agg gac ccc ggt gaa aca gta ggt gga gat gag att 847 Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly Gly Asp Glu He 135 140 145 gtg atg gtc ate aaa gac age cca caa acc ate cca aca tac aca gat 895 Val Met Val He Lys Asp Ser Pro Gin Thr He Pro Thr Tyr Thr Asp 150 155 160 gcc ttg cac gtt tat tec acg gta gag gga tac ate gcc tac cga cat 943 Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He Ala Tyr Arg His 165 170 175 180 gat cag aaa ggc tea tgc ttt ate cag acc ctg gtg gat gtg ttc acg 991 Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val Asp Val Phe Thr 185 190 195 aag agg aaa gga cat ate ttg gaa ctt ctg aca gag gtg acc egg egg 1039 Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu Val Thr Arg Arg 200 205 210 atg gca gaa gca gag ctg gtt caa gaa gga aaa gca agg aaa acg aac 1087 Met Ala Glu Ala Glu Leu. Val Gin Glu Gly Lys Ala Arg Lys Thr Asn 215 220 225 cct gaa ate caa age acc etc egg aaa egg ctg tat ctg cag 1129 Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr Leu Gin 230 235 240 tagaagtaga aagaccagga ggagctttcc ttccagcatt ctttctgtct cacagaaatt 1189 tagaggcagc tcttacctct ccccaagatc ttctgttccc aaggccaaat ggcacccagt 1249 ttcttttcca tcacaccctt catgcaggtc ctcctgtcct tattagagca agccagccaa 1309 aacttagcac aaggcatggt ggcaacatta acatcacctc cctcaggctg gactttctat 1369 ctttattaat gcaaccgaag agacctaaga gtgeattcac ttatcccact ttctgttcct 1429 gtggtcttct ttctcccatg aagcagaaac tggataaagc tcaagatttt ccatagacaa 1489 accaaagccc actcatcccc tcctacccca atccaacctc tgctggctcc tgcatctcac 1549 ttggaggtca aacctcctcc tgaggccaat gcattcccaa cttccagttc tttcctttac 1609 cctggagagt tagtaaggta agaaccattc tttctctcca aaaccactcc tccttggctg 1669 gcaagttggt gtcctaactc cgttctcttc ctagctcatg gcctctctag ataataaagt 1729 tgtctcctcc tttctggatc tcttcctcct aacacccctc ccctgaaacc ctggactctg 1789 ccctctctcc aagaaaatcc atctattcaa etattcttge attcaattac tctaaatgag 1849 agcgtgttgg agctatggca aattccctgt tgtcaccttg etattttgea gacaacataa 1909 tatttaacct ctcataacca gagaggttaa ataatttgtc aaatgeaata cagtaagaca 1969 gaggcaagga caaggtttga cttccagccc agcctctttt ccacaacctg ctaaatcctg 2029 164,326/1 344 atccatctga aaacttttct aattagtgaa gatgactaat aaaaattttc cctatctcca 2089 aggtaggagc tttctggaag tttctagaaa ttttcaataa ccaccagcca aggttacctc 2149 caggtaacct tgcagcacca ggctggaagt cagatcggct tcactatctt ccaactctac 2209 agcctgtatc tctccattcc cagctttgac ctttcctgct caagtaacct acgggcacat 2269 ccagcgtcac taaaaactca gggcttttct tcccggttac tcctccaagc gttccctggt 2329 atcctcaacc tcagatccca ggttcagatt tctgcagtca atctatgacc cctctcttct 2389 tgcatccttc atatgccacc agacaccatg cccagtccag cctgattttg aaacaacttt 2449 catgccggtc ttctcttccc tgacatgtta ctgtccaggc tcaagtcctc agcttctcat 2509 atctgcatct ttgcaaccaa cttcctccct tgcctctctg cttttccatc ccacttttca 2569 tgtgtcctcc ataccatcta taacagtgat ctccctggaa cactcaagaa gacacaacat 2629 accatattat ttaaagacca gggtactgga cagtggctca cacctgtatt cccgactttg 2689 agagtctgaa gcgggaggat cacttgaggc caggagttaa gagaccagcc tgggcaacac 2749 agcaagaccc tgtctctaaa aaaaaaaatt aattaactgg gtatggtggc acatgcctgt 2809 agtcccagct actcaggagg ctgaggtggg aggatgactt gagcccagga gtttgaggct 2869 gcaaggagct atgatcatgc cagtgcatcc cagctctagg tgagacagtg agatccggtc 2929 tccaaaataa atcaatcaat caaataaaga ccaaagtcaa accgcacatc aggatctctc 2989 acacccttcc aattttgcca tctaccagca cttagctaaa cccatctccc atctcttcca 3049 ccatgaattc actctttcaa aaaggctaat gtcttcttac tcacccttgc ctctaagcct 3109 ttgctatcac catttccccc aagctggagg gccctccctc tccctttacc cctcttccac 3169 tacctcccac ccctactttt tccagaaagc catttcctct cttttttctg attgatcctt 3229 ccctctcacc caggattaga tgctggaaat gaccacttct ggagggcagg gaacaagccc 3289 ttaatctgca taatgagtg t tcaataaaca gttgtcaaac tttgaaa 3336 <210> 15 <211> 242 <212> PRT <213> Homo Sapiens <400> 15 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu As Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He. 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin 164,326/1 345 <210> 16 <211> 3336 <212> DNA <213> Homo Sapiens <220> <221> CDS <222> (404) ... (1129) <400> 16 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaa atg age aat ccg 415 Met Ser Asn Pro 1 egg tct ttg gaa gag gag aaa tat gat atg tea ggt gec cgc ctg gec 463 Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala Arg Leu Ala 10 15 20 eta ata ctg tgt gtc acc aaa gec egg gaa ggt tec gaa gaa gac ctg 511 Leu lie Leu Cys Val Thr Lys Ala Arg Glu Gly Ser Glu Glu Asp Leu 30 35 gat get ctg gaa cac atg ttt egg cag ctg aga ttc gaa age acc atg 559 Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu Ser Thr Met 40 45 50 aaa aga gac ccc act gee gag caa ttc cag gaa gag ctg gaa aaa ttc 607 Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu Glu Lys Phe 55 60 65 cag cag gec ate gat tec egg gaa gat ccc gtc agt tgt gec ttc gtg 655 Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser Cys Ala Phe Val 70 75 80 gta etc atg get cac ggg agg gaa ggc ttc etc aag gga gaa gat ggg 703 Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys Gly Glu Asp Gly 85 90 95 100 gag atg gtc aag ctg gag aat etc ttc gag gee ctg aac aac aag aac 751 Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu Asn Asn Lys Asn 105 110 115 tgc cag gec ctg cga get aag ccc aag gtg tac ate ata cag gee tgt 799 Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He He Gin Ala Cys 120 125 130 cga gga gaa caa agg gac ccc ggt gaa aca gta ggt gga gat gag att 847 Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly Gly Asp Glu He 135 140 145 gtg atg gtc ate aaa gac age cca caa acc ate cca aca tac aca gat 895 Val Met Val He Lys Asp Ser Pro Gin Thr He Pro Thr Tyr Thr Asp 150 155 160 164,326/1 346 gcc ttg cac gtt tat tec acg gta gag gga tac ate gee tac cga cat 943 Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He Ala Tyr Arg His 165 170 175 180 gat cag aaa ggc tea tgc ttt ate eag acc etg gtg gat gtg ttc acg 991 Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val Asp Val Phe Thr 185 190 195 aag agg aaa gga cat ate ttg gaa ctt etg aca gag gtg acc egg egg 1039 Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu Val Thr Arg Arg 200 205 210 atg gca gaa gca gag etg gtt caa gaa gga aaa gca agg aaa acg aac 1087 Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala Arg Lys Thr Asn 215 220 225 cct gaa ate caa age acc etc egg aaa egg etg tat etg cag 1129 Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr Leu Gin 230 235 240 tagaagtaga aagaccagga ggagctttcc ttccagcatt ctttctgtct cacagaaatt 1189 tagaggcagc tcttacctct ccccaagatc ttctgttccc aaggccaaat ggcacccagt 1249 ttcttttcca tcacaccctt catgeaggtc ctcctgtcct tattagagca agccagccaa 1309 aacttagcac aaggcatggt ggcaacatta acatcacctc cctcaggctg gactttctat 1369 ctttattaat gcaaccgaag agacctaaga gtgeattcac ttatcccact ttctgttcct 1429 gtggtcttct ttctcccatg aagcagaaac tggataaagc tcaagatttt ccatagacaa 1489 accaaagccc actcatcccc tcctacccca atccaacctc tgctggctcc tgcatctcac 1549 ttggaggtca aacctcctcc tgaggccaat gcattcccaa cttccagttc tttcctttac 1609 cctggagagt tagtaaggta agaaccattc tttctctcca aaaccactcc tccttggctg 1669 gcaagttggt gtcctaactc cgttctcttc ctagctcatg gcctctctag ataataaagt 1729 tgtctcctcc tttctggatc tcttcctcct aacacccctc ccctgaaacc ctggactctg 1789 ccctctctcc aagaaaatcc atctattcaa etattcttge attcaattac tctaaatgag 1849 agcgtgttgg agctatggca aattccctgt tgtcaccttg etattttgea gacaacataa 1909 tatttaacct ctcataacca gagaggttaa ataatttgtc aaatgeaata cagtaagaca 1969 gaggcaagga caaggtttga cttccagccc agectctttt ccacaacctg ctaaatcctg 2029 atccatccga aaacttttct aattagtgaa gatgactaat aaaaattttc cctatctcca 2089 aggtaggagc tttctggaag tttctagaaa ttttcaataa ccaccagcca aggttacctc 2149 caggtaacct tgcagcacca ggctggaagt cagatcggct tcactatctt ccaactctac 2209 agectgtate tctccatccc cagctttgac ctttcctgct caagtaacct aegggcacat 2269 ccagcgtcac taaaaactca gggcttttct tcccggttac tcctccaagc gttccctggt 2329 atcctcaacc tcagatccca ggttcagatt tetgeagtea atctatgacc cctctcttct 2389 tgcatccttc atatgccacc agacaccatg cccagtccag cctgattttg aaacaacttt 2449 catgccggtc ttctcttccc tgacatgtta ctgtccaggc tcaagtcctc agcttctcat 2509 atetgeatet ttgcaaccaa cttcctccct tgcctctctg cttttccatc ccacttttca 2569 tgtgtcctcc ataccatcta taacagtgat ctccctggaa cactcaagaa gacacaacat 2629 accatattat ttaaagacca gggtactgga cagtggctca cacctgtatt cccgactttg 2689 agagtctgaa gegggaggat cacttgaggc caggagttaa gagaccagcc tgggcaacac 2749 agcaagaccc tgtctctaaa aaaaaaaatt aattaactgg gtatggtggc acatgectgt 2809 agtcccagct actcaggagg ctgaggtggg aggatgactt gageccagga gtttgaggct 2869 gcaaggagct atgatcatgc cagtgcatcc cagctctagg tgagacagtg agatceggtc 2929 tccaaaataa atcaatcaat caaataaaga ccaaagtcaa accgcacatc aggatctctc 2989 acacccttcc aattttgeca tctaccagca cttagctaaa cccatctccc atctcttcca 3049 ccatgaattc actctttcaa aaaggctaat gtcttcttac tcacccttgc ctctaagcct 3109 ttgetatcac catttccccc aagctggagg gccctccctc tccctttacc cctcttccac 3169 tacctcccac ccctactttt tccagaaagc catttcctct cttttttctg attgatcctt 3229 ccctctcacc caggattaga tgctggaaat gaccacttct ggagggcagg gaacaagccc 3289 ttaatctgea taatgagtgt tcaataaaca gttgtcaaac tttgaaa 3336 <210> 17 <211> 242 <212> PRT <213> Homo Sapiens <400> 17 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 18 <211> 242 <212> PRT <213> Homo Sapiens <400> 18 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 19 <211> 230 <212> PRT <213> Homo Sapiens <400> 19 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Pro Thr 165 170 175 Pro Phe Gin Asp Pro Leu Tyr Leu Pro Ser Glu Ala Pro Pro Asn Pro 180 185 190 Pro Leu Trp Asn Ser Gin Asp Thr Ser Pro Thr Asp Met He Arg Lys 195 200 205 Ala His Ala Leu Ser Arg Pro Trp Trp Met Cys Ser Arg Arg Gly Lys 210 215 220 Asp He Ser Trp Asn Phe 225 230 <210> 20 <211> 146 <212> PRT <213> Homo Sapiens <400> 20 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Ala Thr Leu Pro Ser Pro Phe Pro Tyr Leu 130 135 140 Ser Leu 145 <210> 21 <211> 321 <212> PRT <213> Homo Sapiens <400> 21 Met Gly Lys Cys Gin Glu Tyr Asp Lys Ser Leu Ser Val Gin Pro Glu 1 5 10 15 Lys Arg Thr Gly Leu Arg Asp Glu Asn Gly Glu Cys Gly Gin Thr Phe 25 30 Arg Leu Lys Glu Glu Gin Gly Arg Ala Phe Arg Gly Ser Ser Val His 40 45 Gin Lys Leu Val Asn Asp Pro Arg Glu Thr Gin Glu Val Phe Gly Gly 50 55 60 Gly Val Gly Asp He Val Gly Arg Asp Leu Ser He Ser Phe Arg Asn 65 70 75 80 Ser Glu Thr Ser Ala Ser Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala 85 90 95 Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser Glu 100 105 110 Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu 115 120 125 Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu 130 135 140 Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser Cys 145 150 155 160 Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys Gly 165 170 175 Glu Asp Gly Glu Met Val Lys Leu Glu As Leu Phe Glu Ala Leu Asn 180 185 190 Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He He 195 200 205 Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly Gly 210 215 220 Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro Thr 225 230 235 240 Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He Ala 245 250 255 Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val Asp 260 265 270 Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu Val 275 280 285 Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala Arg 290 295 300 Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr Leu 305 310 315 320 Gin <210> 22 <211> 242 <212> PRT <213> Homo Sapiens <400> 22 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Arg Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg. His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 23 <211> 242 <212> PRT <213> Homo Sapiens <400> 23 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 · 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 164,326/1 351 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Met 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 24 <211> 776 <212> DNA <213> Homo Sapiens <400> 24 ggatcagaca agggtgctga gagccgggac tcacaaccaa aggagaaatg agcaatccgc 60 ggtctttgga agaggagaaa tatgatatgt caggtgcccg cctggcccta atactgtgtg 120 tcaccaaagc ccgggaaggt tccgaagaag acctggatgc tctggaacac atgtttcggc 180 agctgagatt cgaaagcacc atgaaaagag accccactgc cgagcaattc caggaagagc 240 tggaaaaatt ccagcaggcc atcgattccc gggaagatcc cgtcagttgt gccttcgtgg 300 tactcatggc tcacgggagg gaaggcttcc tcaagggaga agatggggag atggtcaagc 360 tggagaatct cttcgaggcc ctgaacaaca agaactgcca ggccctgcga gctaagccca 420 aggtgtacat catacaggcc tgtcgaggag aacaaaggga ccccggtgaa acagtaggtg 480 gagatgagat tgtgatggtc atcaaagaca gcccacaaac catcccaaca tacacagatg 540 ccttgcacgt ttattccacg gtagagggat acatcgccta ccgacatgat cagaaaggct 600 catgctttat ccagaccctg gtggatgtgt tcacgaagag gaaaggacat atcttggaac 660 ttctgacaga ggtgacccgg cggatggcag aagcagagct ggttcaagaa ggaaaagcaa 720 ggaaaacgaa ccctgaaatc caaagcaccc tccggaaacg gctgtatctg cagtag 776 <210> 25 <211> 776 <212> DNA <213> Homo Sapiens <400> 25 ggatcagaca agggtgctga gagccgggac tcacaaccaa aggagaaatg agcaatccgc 60 ggtctttgga agaggagaaa tatgatatgt caggtgcccg cctggcccta atactgtgtg 120 tcaccaaagc ccgggaaggt tccgaagaag acctggatgc tctggaacac atgtttcggc 180 agctgagatt cgaaagcacc atgaaaagag accccactgc cgagcaattc caggaagagc 240 tggaaaaatt ccagcaggcc atcgattccc gggaagatcc cgtcagttgt gccttcgtgg 300 tactcatggc tcacgggagg gaaggcttcc tcaagggaga agatggggag atggtcaagc 360 164,326/1 352 tggagaatct cttcgaggcc ctgaacaaca agaactgcca ggccctgcga gctaagccca 420 aggtgtacat catacaggcc tgtcgaggag aacaaaggga ccccggtgaa acagtaggtg 480 gagatgagat tgtgatggtc atcaaagaca gcccacaaac catcccaaca tacacagatg 540 ccttgcacgt ttattccacg gtagagggat acatcgccta ccgacatgat cagaaaggct 600 catgctttat ccagaccctg gtggatgtgt tcacgaagag gaaaggacat atcttggaac 660 ttctgacaga ggtgacccgg cggatggcag aagcagagct ggttcaagaa ggaaaagcaa 720 ggaaaacgaa ccctgaaatc caaagcaccc tccggaaacg gctgtatctg cagtag 776 <210> 26 <211> 242 <212> PRT <213> Homo Sapiens <400> 26 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 27 <211> 242· <212> PRT <213> Homo Sapiens <400> 27 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 28 <2H> 242 <212> PRT <213> Homo Sapiens <400> 28 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5' 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 29 <211> 253 <212> PRT <213> Mus musculus <400> 29 Met Ser Asp Pro Gin Pro Leu Gin Glu Glu Arg Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu Thr Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Val Asp Met Glu Ala Leu Glu Arg Met Phe Arg Tyr Leu Lys Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Gin Gin Phe Leu Glu Glu 50 55 60 Leu Asp Glu Phe Gin Gin Thr He Asp Asn Trp Glu Glu Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Glu Glu Gly Leu Leu Lys 85 90 95 Gly Glu Asp Glu Lys Met Val Arg Leu Glu Asp Leu Phe Glu Val Leu 100 105 110 Asn Asn Lys Asn Cys Lys Ala Leu Arg Gly Lys Pro Lys Val Tyr He 115 120 125 lie Gin Ala Cys Arg Gly Glu His Arg Asp Pro Gly Glu Glu Leu Arg 130 135 140 Gly Asn Glu Glu Leu Gly Gly Asp Glu Glu Leu Gly Gly Asp Glu Val 145 150 155 160 Ala Val Leu Lys Asn Asn Pro Gin Ser He Pro Thr Tyr Thr Asp Thr 165 170 175 Leu His He Tyr Ser Thr Val Glu Gly Tyr Leu Ser Tyr Arg His Asp 180 185 190 Glu Lys Gly Ser Gly Phe He Gin Thr Leu Thr Asp Val Phe He His 195 200 205 Lys Lys Gly Ser He Leu Glu Leu Thr Glu Glu He Thr Arg Leu Met 210 215 220 Ala Asn Thr Glu Val Met Gin Glu Gly Lys Pro Arg Lys Val Asn Pro 225 230 235 240 Glu Val Gin Ser Thr Leu Arg Lys Lys Leu Tyr Leu Gin 245 250 <210> 30 <211> 242 <212> PRT <213> Homo : Sapiens <400> 30 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 31 <211> 242 <212> PRT <213> Homo Sapiens <400> 31 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu, Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 32 <211> 134 <212> PRT <213> Homo Sapiens <400> 32 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly 130 <210> 33 <211> 134 <212> PRT <213> Homo Sapiens <400> 33 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly 130 <210> 34 <211> 174 <212> PRT <213> Homo Sapiens <400> 34 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly 165 170 <210> 35 <211> 185 <212> PRT <213> Mus musculus <400> 35 Met Ser Asp Pro Gin Pro Leu Gin Glu Glu Arg Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu Thr Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Val Asp Met Glu Ala Leu Glu Arg Met Phe Arg Tyr Leu Lys Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Gin Gin Phe Leu Glu Glu 50 55 60 Leu Asp Glu Phe Gin Gin Thr He Asp Asn Trp Glu Glu Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Glu Glu Gly Leu Leu Lys 85 90 95 Gly Glu Asp Glu Lys Met Val Arg Leu Glu Asp Leu Phe Glu Val Leu 100 105 110 Asn Asn Lys Asn Cys Lys Ala Leu Arg Gly Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu His Arg Asp Pro Gly Glu Glu Leu Arg 130 135 140 Gly Asn Glu Glu Leu Gly Gly Asp Glu Glu Leu Gly Gly Asp Glu Val 145 150 155 160 Ala Val Leu Lys Asn Asn Pro Gin Ser He Pro Thr Tyr Thr Asp Thr 165 170 175 Leu His He Tyr Ser Thr Val Glu Gly 180 185 <210> 36 <211> 241 <212> PRT <213> Homo Sapiens <400> 36 Ser Glu Thr Ser Ala Ser Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala 1 5 10 15 Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser Glu 25 30 Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu 40 45 Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu 50 55 60 Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser Cys 65 70 75 80 Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys Gly 85 90 95 Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu Asn 100 105 110 Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He He 115 120 125 Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly Gly 130 135 140 Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro Thr 145 150 155 160 Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He Ala 165 170 175 Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val Asp 180 185 190 Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu Val 195 200 205 Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala Arg 210 215 220 Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr Leu 225 230 235 240 Gin <210> 37 <211> 241 <212> PRT <213> Homo Sapiens <400> 37 Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala 1 5 10 15 Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser Glu 25 30 Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu 40 45 Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu 50 55 60 Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser Cys 65 70 75 80 Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys Gly 85 90 95 Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu Asn 100 105 110 Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He He 115 120 125 Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly Gly 130 135 140 Asp Glu He Val Met Val lie Lys Asp Ser Pro Gin Thr He Pro Thr 145 150 155 160 Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr lie Ala 165 170 175 Tyr Arg His Asp Gin Lys Gly Ser Cys Phe lie Gin Thr Leu Val Asp 180 185 190 Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu Val 195 200 205 Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala Arg 210 215 220 Lys Thr Asn Pro Glu lie Gin Ser Thr Leu Arg Lys Arg Leu Tyr Leu 225 230 235 240 Gin <210> 38 <211> 14 <212> PRT <213> Clostridiumn toxi <220> <223> Artificially Synthesized Peptide <400> 38 Gin Tyr He Lys Ala Asn Ser Lys Phe He Gly He Thr Glu 1 5 10 <210> 39 <211> 21 <212> PRT <213> Plasmodium falciparum <400> 39 Asp He Glu Lys Lys He Ala Lys Met Glu Lys Ala Ser Ser Val Phe 1 5 10 15 Asn Val Val Asn Ser <210> 40 <211> 16 <212> PRT <213> Streptococcus Aureus <400> 40 Gly Ala Val Asp Ser He Leu Gly Gly Val Ala Thr Tyr Gly Ala Ala 1 5 10 15 <210> 41 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Artificially Synthesized Peptide <221> VARIANT <222> 3 <223> Xaa = cyclohexylalanine, phenylalanine, or tyrosine <221> VARIANT <222> 1, 13 <223> Xaa = D-alanine or L-alanine <400> 41 Xaa Lys Xaa Val Ala Ala Trp Thr Leu Lys Ala Ala 1 5 10 <210> 42 <211> 43 <212> DNA <213> Artificial Sequence <220> <223> Artificially Synthesized Primer <400> 42 ttttgatcaa gctttttttt tttttttttt tttttttttt ttt <210> 43 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> Artificially Synthesized Primer <400> 43 ctaatacgac tcactatagg gctcgagcgg ccgcccgggc ag <210> 44 <211> 12 <212> DNA <213> Artificial Sequence <220> <223> Artificially Synthesized Primer <400> 44 gatcctgccc gg <210> 45 <211> 40 <212> DNA <213> Artificial Sequence <220> <223> Artificially Synthesized Primer <400> 45 gtaatacgac tcactatagg gcagcgtggt cgcggccgag <210> 46 <211> 10 <212> DNA <213> Artificial Sequence <220> <223> Artificially Synthesized Primer <400> 46 gatcctcggc <210> 47 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Artificially Synthesized Primer <400> 47 ctaatacgac tcactatagg gc <210> 48 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Artificially Synthesized Primer <400> 48 tcgagcggcc gcccgggcag ga <210> 49 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Artificially Synthesized Primer <400> 49 agcgtggtcg cggccgagga <210> 50 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> Artificially Synthesized Primer <400> 50 atatcgccgc gctcgtcgtc gacaa <210> 51 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> Artificially Synthesized Primer <400> 51 agccacacgc agctcattgt agaagg <210> 52 <211> 22 <212> DNA 164,326/1 362 <213> Artificial Sequence <220> <223> Artificially Synthesized Primer <400> 52 ggataccagg gaacgcttgg ag 22 <210> 53 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> Artificially Synthesized Primer <400> 53 tttgaccttt cctgctcaag taacc 25 <210> 54 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Artificially Synthesized Primer <400> 54 gattacaagg atgacgacga taag 24 <210> 55 <211> 3410 <212> DNA <213> Homo Sapiens <400> 55 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaaatgagca atccgcggtc 420 tttggaagag gagaaatatg atatgtcagg tgcccgcctg gccctaatac tgtgtgtcac 480 caaagcccgg gaaggttccg aagaagacct ggatgctctg gaacacatgt ttcggcagct 540 gagattcgaa agcaccatga aaagagaccc cactgccgag caattccagg aagagctgga 600 aaaattccag caggccatcg attcccggga agatcccgtc agttgtgcct tcgtggtact 660 catggctcac gggagggaag gcttcctcaa gggagaagat ggggagatgg tcaagctgga 720 gaatctcttc gaggccctga acaacaagaa ctgccaggcc ctgcgagcta agcccaaggt 780 gtacatcata caggcctgtc gaggagaaca aagggacccc ggtgaaacag taggtggaga 840 tgagattgtg atggtcatca aagacagccc acaaaccatc ccaacataca cagatgcctt 900 gcacgtttat tccacggtag agggacccac gcccttccag gatcccctct acctaccctc 960 tgaagctccc ccgaacccac ctctctggaa ttcccaggat acatcgccta ccgacatgat 1020 cagaaaggct catgctttat ccagaccctg gtggatgtgt tcacgaagag gaaaggacat 1080 atcttggaac ttctgacaga ggtgacccgg cggatggcag aagcagagct ggttcaagaa 1140 ggaaaagcaa ggaaaacgaa ccctgaaatc caaagcaccc tccggaaacg gctgtatctg 1200 cagtagaagt agaaagacca ggaggagctt tccttccagc attctttctg tctcacagaa 1260 atttagaggc agctcttacc tctccccaag atcttctgtt cccaaggcca aatggcaccc 1320 agtttctttt ccatcacacc cttcatgcag gtcctcctgt ccttattaga gcaagccagc 1380 caaaacttag cacaaggcat ggtggcaaca ttaacatcac ctccctcagg ctggactttc 1440 tatctttatt aatgcaaccg aagagaccta agagtgcatt cacttatccc actttctgtt 1500 164,326/1 363 cctgtggtct tctttctccc atgaagcaga aactggataa agctcaagat tttccataga 1560 caaaccaaag cccactcatc ccctcctacc ccaatccaac ctctgctggc tcctgcatct 1620 cacttggagg tcaaacctcc tcctgaggcc aatgcattcc caacttccag ttctttcctt 1680 taccctggag agttagtaag gtaagaacca ttctttctct ccaaaaccac tcctccttgg 1740 ctggcaagtt ggtgtcctaa ctccgttctc ttcctagctc atggcctctc tagataataa 1800 agttgtctcc tcctttctgg atctcttcct cctaacaccc ctcccctgaa accctggact 1860 ctgccctctc tccaagaaaa tccatctatt caactattct tgcattcaat tactctaaat 1920 gagagcgtgt tggagctatg gcaaattccc tgttgtcacc ttgctatttt gcagacaaca 1980 taatatttaa cctctcataa ccagagaggt taaataattt gtcaaatgca atacagtaag 2040 acagaggcaa ggacaaggtt tgacttccag cccagcctct tttccacaac ctgctaaatc 2100 ctgatccatc tgaaaacttt tctaattagt gaagatgact aataaaaatt ttccctatct 2160 ccaaggtagg agctttctgg aagtttctag aaattttcaa taaccaccag ccaaggttac 2220 ctccaggtaa ccttgcagca ccaggctgga agtcagatcg gcttcactat cttccaactc 2280 tacagcctgt atctctccat ccccagcttt gacctttcct gctcaagtaa cctacgggca 2340 catccagcgt cactaaaaac tcagggcttt tcttcccggt tactcctcca agcgttccct 2400 ggtatcctca acctcagatc ccaggttcag atttctgcag tcaatctatg acccctctct 2460 tcttgcatcc ttcatatgcc accagacacc atgcccagtc cagcctgatt ttgaaacaac 2520 tttcatgccg gtcttctctt ccctgacatg ttactgtcca ggctcaagtc ctcagcttct 2580 catatctgca tctttgcaac caacttcctc ccttgcctct ctgcttttcc atcccacttt 2640 tcatgtgtcc tccataccat ctataacagt gatctccctg gaacactcaa gaagacacaa 2700 cataccatat tatttaaaga ccagggtact ggacagtggc tcacacctgt attcccgact 2760 ttgagagtct gaagcgggag gatcacttga ggccaggagt taagagacca gcctgggcaa 2820 cacagcaaga ccctgtctct aaaaaaaaaa attaattaac tgggtatggt ggcacatgcc 2880 tgtagtccca gctactcagg aggctgaggt gggaggatga cttgagccca ggagtttgag 2940 gctgcaagga gctatgatca tgccagtgca tcccagctct aggtgagaca gtgagatccg 3000 gtctccaaaa taaatcaatc aatcaaataa agaccaaagt caaaccgcac atcaggatct 3060 ctcacaccct tccaattttg ccatctacca gcacttagct aaacccatct cccatctctt 3120 ccaccatgaa ttcactcttt caaaaaggct aatgtcttct tactcaccct tgcctctaag 3180 cctttgctat caccatttcc cccaagctgg agggccctcc ctctcccttt acccctcttc 3240 cactacctcc cacccctact ttttccagaa agccatttcc tctctttttt ctgattgatc 3300 cttccctctc acccaggatt agatgctgga aatgaccact tctggagggc agggaacaag 3360 cccttaatct gcataatgag tgttcaataa acagttgtca aactttgaaa 3410 <210> 56 <211> 3404 <212> DNA <213> Homo Sapiens <400> 56 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaaatgagca atccgcggtc 420 tttggaagag gagaaatatg atatgtcagg tgcccgcctg gccctaatac tgtgtgtcac 480 caaagcccgg gaaggttccg aagaagacct ggatgctctg gaacacatgt ttcggcagct 540 gagattcgaa agcaccatga aaagagaccc cactgccgag caattccagg aagagctgga 600 aaaattccag caggccatcg attcccggga agatcccgtc agttgtgcct tcgtggtact 660 catggctcac gggagggaag gcttcctcaa gggagaagat ggggagatgg tcaagctgga 720 gaatctcttc gaggccctga acaacaagaa ctgccaggcc ctgcgagcta agcccaaggt 780 gtacatcata caggcctgtc gaggagccac cctgcccagc ccctttcctt acctttctct 840 ctgactttgc ctcctcctct tcttgttgtt tcagaacaaa gggaccccgg tgaaacagta 900 ggtggagatg agattgtgat ggtcatcaaa gacagcccac aaaccatccc aacatacaca 960 gatgccttgc acgtttattc cacggtagag ggatacatcg cctaccgaca tgatcagaaa 1020 ggctcatgct ttatccagac cctggtggat gtgttcacga agaggaaagg acatatcttg 1080 gaacttctga cagaggtgac ccggcggatg gcagaagcag agctggttca agaaggaaaa 1140 gcaaggaaaa cgaaccctga aatccaaagc accctccgga aacggctgta tctgcagtag 1200 aagtagaaag accaggagga gctttccttc cagcattctt tctgtctcac agaaatttag 1260 aggcagctct tacctctccc caagatcttc tgttcccaag gccaaatggc acccagtttc 1320 164,326/1 364 ttttccatca cacccttcat gcaggtcctc ctgtccttat tagagcaagc cagccaaaac 1380 ttagcacaag gcatggtggc aacattaaca tcacctccct caggctggac tttctatctt 1440 tattaatgca accgaagaga cctaagagtg cattcactta tcccactttc tgttcctgtg 1500 gtcttctttc tcccatgaag cagaaactgg ataaagctca agattttcca tagacaaacc 1560 aaagcccact catcccctcc taccccaatc caacctctgc tggctcctgc atctcacttg 1620 gaggtcaaac ctcctcctga ggccaatgca ttcccaactt ccagttcttt cctttaccct 1680 ggagagttag taaggtaaga accattcttt ctctccaaaa ccactcctcc ttggctggca 1740 agttggtgtc ctaactccgt tctcttccta gctcatggcc tctctagata ataaagttgt 1800 ctcctccttt ctggatctct tcctcctaac acccctcccc tgaaaccctg gactctgccc 1860 tctctccaag aaaatccatc tattcaacta ttcttgcatt caattactct aaatgagagc 1920 gtgttggagc tatggcaaat tccctgttgt caccttgcta ttttgcagac aacataatat 1980 ttaacctctc ataaccagag aggttaaata atttgtcaaa tgcaatacag taagacagag 2040 gcaaggacaa ggtttgactt ccagcccagc ctcttttcca caacctgcta aatcctgatc 2100 catctgaaaa cttttctaat tagtgaagat gactaataaa aattttccct atctccaagg 2160 taggagcttt ctggaagttt ctagaaattt tcaataacca ccagccaagg ttacctccag 2220 gtaaccttgc agcaccaggc tggaagtcag atcggcttca ctatcttcca actctacagc 2280 ctgtatctct ccatccccag ctttgacctt tcctgctcaa gtaacctacg ggcacatcca 2340 gcgtcactaa aaactcaggg cttttcttcc cggttactcc tccaagcgtt ccctggtatc 2400 ctcaacctca gatcccaggt tcagatttct gcagtcaatc tatgacccct ctcttcttgc 2460 atccttcata tgccaccaga caccatgccc agtccagcct gattttgaaa caactttcat 2520 gccggtcttc tcttccctga catgttactg tccaggctca agtcctcagc ttctcatatc 2580 tgcatctttg caaccaactt cctcccttgc ctctctgctt ttccatccca cttttcatgt 2640 gtcctccata ccatctataa cagtgatctc cctggaacac tcaagaagac acaacatacc 2700 atattattta aagaccaggg tactggacag tggctcacac ctgtattccc gactttgaga 2760 gtctgaagcg ggaggatcac ttgaggccag gagttaagag accagcctgg gcaacacagc 2820 aagaccctgt ctctaaaaaa aaaaattaat taactgggta tggtggcaca tgcctgtagt 2880 cccagctact caggaggctg aggtgggagg atgacttgag cccaggagtt tgaggctgca 2940 aggagctatg atcatgccag tgcatcccag ctctaggtga gacagtgaga tccggtctcc 3000 aaaataaatc aatcaatcaa ataaagacca aagtcaaacc gcacatcagg atctctcaca 3060 cccttccaat tttgccatct accagcactt agctaaaccc atctcccatc tcttccacca 3120 tgaattcact ctttcaaaaa ggctaatgtc ttcttactca cccttgcctc taagcctttg 3180 ctatcaccat ttcccccaag ctggagggcc ctccctctcc ctttacccct cttccactac 3240 ctcccacccc tactttttcc agaaagccat ttcctctctt ttttctgatt gatccttccc 3300 tctcacccag gattagatgc tggaaatgac cacttctgga gggcagggaa caagccctta 3360 atctgcataa tgagtgttca ataaacagtt gtcaaacttt gaaa 3404 <210> 57 <211> 966 <212> DNA <213> Homo Sapiens <400> 57 atggggaaat gccaagagta tgacaaaagt ctgtctgtgc agccagagaa gagaacagga 60 ctcagagatg agaatggaga atgtggacag acattcagac tcaaggaaga gcaagggagg 120 gctttcaggg gaagttcagt ccaccagaag ctggtgaatg acccacggga gacacaggaa 180 gtttttgggg gcggagtggg ggacattgtg ggacgggatc tcagtattag cttcagaaac 240 tctgagacct ctgcaagtga ggaggagaaa tatgatatgt caggtgcccg cctggcccta 300 atactgtgtg tcaccaaagc ccgggaaggt tccgaagaag acctggatgc tctggaacac 360 atgtttcggc agctgagatt cgaaagcacc atgaaaagag accccactgc cgagcaattc 420 caggaagagc tggaaaaatt ccagcaggcc atcgattccc gggaagatcc cgtcagttgt 480 gccttcgtgg tactcatggc tcacgggagg gaaggcttcc tcaagggaga agatggggag 540 atggtcaagc tggagaatct cttcgaggcc ctgaacaaca agaactgcca ggccctgcga 600 gctaagccca aggtgtacat catacaggcc tgtcgaggag aacaaaggga ccccggtgaa 660 acagtaggtg gagatgagat tgtgatggtc atcaaagaca gcccacaaac catcccaaca 720 tacacagatg ccttgcacgt ttattccacg gtagagggat acatcgccta ccgacatgat 780 cagaaaggct catgctttat ccagaccctg gtggatgtgt tcacgaagag gaaaggacat 840 atcttggaac ttctgacaga ggtgacccgg cggatggcag aagcagagct ggttcaagaa 900 ggaaaagcaa ggaaaacgaa ccctgaaatc caaagcaccc tccggaaacg gctgtatctg 960 cagtag 966 <210> 58 164,326/1 365 <211> 3336 <212> DNA <213> Homo Sapiens <400> 58 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaaatgagca atccgcggtc 420 tttggaagag gagaaatatg atatgtcagg tgcccgcctg gccctaatac tgtgtgtcac 480 caaagcccgg gaaggttccg aagaagacct ggatgctctg gaacacatgt ttcggcagct 540 gagattcgaa agcaccatga aaagagaccc cactgccgag caattccagg aagagctgga 600 aaaattccag caggccatcg attcccggga agatcccgtc agttgtgcct tcgtggtact 660 catggctcac gggagggaag gcttcctcaa gggagaagat ggggagatgg tcaagctgga 720 gaatctcttc gaggccctga acaacaagaa ctgccaggcc ctgcgagcta agcccaaggt 780 gtacatcata caggcctgtc gaggagaaca aagggacccc ggtgaaacag taggtggaga 840 tgagattgtg atggtcatca aagacagccc acaaaccatc ccaacataca cagatgcctt 900 gcacgtttat tccacggtag agggatacat cgcctaccga catgatcaga aaggctcatg 960 ctttatccag accctggtgg atgtgttcac gaagaggaaa ggacatatct tggaacttct 1020 gacagaggtg acccggcgga tggcagaagc agagctggtt caagaaggaa aagcaaggaa 1080 aacgaaccct gaaatccaaa gcaccctccg gaaacggctg tatctgcagt agaagtagaa 1140 agaccaggag gagctttcct tccagcattc tttctgtctc acagaaattt agaggcagct 1200 cttacctctc cccaagatct tctgttccca aggccaaatg gcacccagtt tcttttccat 1260 cacacccttc atgcaggtcc tcctgtcctt attagagcaa gccagccaaa acttagcaca 1320 aggcatggtg gcaacattaa catcacctcc ctcaggctgg actttctatc tttattaatg 1380 caaccgaaga gacctaagag tgcattcact tatcccactt tctgttcctg tggtcttctt 1440 tctcccatga agcagaaact ggataaagct caagattttc catagacaaa ccaaagccca 1500 ctcatcccct cctaccccaa tccaacctct gctggctcct gcatctcact tggaggtcaa 1560 acctcctcct gaggccaatg cattcccaac ttccagttct ttcctttacc ctggagagtt 1620 agtaaggtaa gaaccattct ttctctccaa aaccactcct ccttggctgg caagttggtg 1680 tcctaactcc gttctcttcc tagctcatgg cctctctaga taataaagtt gtctcctcct 1740 ttctggatct cttcctccta acacccctcc cctgaaaccc tggactctgc cctctctcca 1800 agaaaatcca tctattcaac tattcttgca ttcaattact ctaaatgaga gcgtgttgga 1860 gctatggcaa attccctgtt gtcaccttgc tattttgcag acaacataat atttaacctc 1920 tcataaccag agaggttaaa taatttgtca aatgcaatac agtaagacag aggcaaggac 1980 aaggtttgac ttccagccca gcctcttttc cacaacctgc taaatcctga tccatctgaa 2040 aacttttcta attagtgaag atgactaata aaaattttcc ctatctccaa ggtaggagct 2100 ttctggaagt ttctagaaat tttcaataac caccagccaa ggttacctcc aggtaacctt 2160 gcagcaccag gctggaagtc agatcggctt cactatcttc caactctaca gcctgtatct 2220 ctccatcccc agctttgacc tttcctgctc aagtaaccta cgggcacatc cagcgtcact 2280 aaaaactcag ggcttttctt cccggttact cctccaagcg ttccctggta tcctcaacct 2340 cagatcccag gttcagattt ctgcagtcaa tctatgaccc ctctcttctt gcatccttca 2400 tatgccacca gacaccatgc ccagtccagc ctgattttga aacaactttc atgccggtct 2460 tctcttccct gacatgttac tgtccaggct caagtcctca gcttctcata tctgcatctt 2520 tgcaaccaac ttcctccctt gcctctctgc ttttccatcc cacttttcat gtgtcctcca 2580 taccatctat aacagtgatc tccctggaac actcaagaag acacaacata ccatattatt 2640 taaagaccag ggtactggac agtggctcac acctgtattc ccgactttga gagtctgaag 2700 cgggaggatc acttgaggcc aggagttaag agaccagcct gggcaacaca gcaagaccct 2760 gtctctaaaa aaaaaaatta attaactggg tatggtggca catgcctgta gtcccagcta 2820 ctcaggaggc tgaggtggga ggatgacttg agcccaggag tttgaggctg caaggagcta 2880 tgatcatgcc agtgcatccc agctctaggt gagacagtga gatccggtct ccaaaataaa 2940 tcaatcaatc aaataaagac caaagtcaaa ccgcacatca ggatctctca cacccttcca 3000 attttgccat ctaccagcac ttagctaaac ccatctccca tctcttccac catgaattca 3060 ctctttcaaa aaggctaatg tcttcttact cacccttgcc tctaagcctt tgctatcacc 3120 atttccccca agctggaggg ccctccctct ccctttaccc ctcttccact acctcccacc 3180 cctacttttt ccagaaagcc atttcctctc ttttttctga ttgatccttc cctctcaccc 3240 aggattagat gctggaaatg accacttctg gagggcaggg aacaagccct taatctgcat 3300 aatgagtgtt caataaacag ttgtcaaact ttgaaa 3336 164,326/1 366 <210> 59 <211> 3410 <212> DNA <213> Homo Sapiens <400> 59 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaaatgagca atccgcggtc 420 tttggaagag gagaaatatg atatgtcagg tgcccgcctg gccctaatac tgtgtgtcac 480 caaagcccgg gaaggttccg aagaagacct ggatgctctg gaacacatgt ttcggcagct 540 gagattcgaa agcaccatga aaagagaccc cactgccgag caattccagg aagagctgga 600 aaaattccag caggccatcg attcccggga agatcccgtc agttgtgcct tcgtggtact 660 catggctcac gggagggaag gcttcctcaa gggagaagat ggggagatgg tcaagctgga 720 gaatctcttc gaggccctga acaacaagaa ctgccaggcc ctgcgagcta agcccaaggt 780 gtacatcata caggcctgtc gaggagaaca aagggacccc ggtgaaacag taggtggaga 840 tgagattgtg atggtcatca aagacagccc acaaaccatc ccaacataca cagatgcctt 900 gcacgtttat tccacggtag agggacccac gcccttccag gatcccctct acctaccctc 960 tgaagctccc ccgaacccac ctctctggaa ttcccaggat acatcgccta ccgacatgat 1020 cagaaaggct catgctttat ccagaccctg gtggatgtgt tcacgaagag gaaaggacat 1080 atcttggaac ttctgacaga ggtgacccgg cggatggcag aagcagagct ggttcaagaa 1140 ggaaaagcaa ggaaaacgaa ccctgaaatc caaagcaccc tccggaaacg gctgtatctg 1200 cagtagaagt agaaagacca ggaggagctt tccttccagc attctttctg tctcacagaa 1260 atttagaggc agctcttacc tctccccaag atcttctgtt cccaaggcca aatggcaccc 1320 agtttctttt ccatcacacc cttcatgcag gtcctcctgt ccttattaga gcaagccagc 1380 caaaacttag cacaaggcat ggtggcaaca ttaacatcac ctccctcagg ctggactttc 1440 tatctttatt aatgcaaccg aagagaccta agagtgcatt cacttatccc actttctgtt 1500 cctgtggtct tctttctccc atgaagcaga aactggataa agctcaagat tttccataga 1560 caaaccaaag cccactcatc ccctcctacc ccaatccaac ctctgctggc tcctgcatct 1620 cacttggagg tcaaacctcc tcctgaggcc aatgcattcc caacttccag ttctttcctt 1680 taccctggag agttagtaag gtaagaacca ttctttctct ccaaaaccac tcctccttgg 1740 ctggcaagtt ggtgtcctaa ctccgttctc ttcctagctc atggcctctc tagataataa 1800 agttgtctcc tcctttctgg atctcttcct cctaacaccc ctcccctgaa accctggact 1860 ctgccctctc tccaagaaaa tccatctatt caactattct tgcattcaat tactctaaat 1920 gagagcgtgt tggagctatg gcaaattccc tgttgtcacc ttgctatttt gcagacaaca 1980 taatatttaa cctctcataa ccagagaggt taaataattt gtcaaatgca atacagtaag 2040 acagaggcaa ggacaaggtt tgacttccag cccagcctct tttccacaac ctgctaaatc 2100 ctgatccatc tgaaaacttt tctaattagt gaagatgact aataaaaatt ttccctatct 2160 ccaaggtagg agctttctgg aagtttctag aaattttcaa taaccaccag ccaaggttac 2220 ctccaggtaa ccttgcagca ccaggctgga agtcagatcg gcttcactat cttccaactc 2280 tacagcctgt atctctccat ccccagcttt gacctttcct gctcaagtaa cctacgggca 2340 catccagcgt cactaaaaac tcagggcttt tcttcccggt tactcctcca agcgttccct 2400 ggtatcctca acctcagatc ccaggttcag atttctgcag tcaatctatg acccctctct 2460 tcttgcatcc ttcatatgcc .accagacacc atgcccagtc cagcctgatt ttgaaacaac 2520 tttcatgccg gtcttctctt ccctgacatg ttactgtcca ggctcaagtc ctcagcttct 2580 catatctgca tctttgcaac caacttcctc ccttgcctct ctgcttttcc atcccacttt 2640 tcatgtgtcc tccataccat ctataacagt gatctccctg gaacactcaa gaagacacaa 2700 cataccatat tatttaaaga ccagggtact ggacagtggc tcacacctgt attcccgact 2760 ttgagagtct gaagcgggag gatcacttga ggccaggagt taagagacca gcctgggcaa 2820 cacagcaaga ccctgtctct aaaaaaaaaa attaattaac tgggtatggt ggcacatgcc 2880 tgtagtccca gctactcagg aggctgaggt gggaggatga cttgagccca ggagtttgag 2940 gctgcaagga gctatgatca tgccagtgca tcccagctct aggtgagaca gtgagatccg 3000 gtctccaaaa taaatcaatc aatcaaataa agaccaaagt caaaccgcac atcaggatct 3060 ctcacaccct tccaattttg ccatctacca gcacttagct aaacccatct cccatctctt 3120 ccaccatgaa ttcactcttt caaaaaggct aatgtcttct tactcaccct tgcctctaag 3180 cctttgctat caccatttcc cccaagctgg agggccctcc ctctcccttt acccctcttc 3240 cactacctcc cacccctact ttttccagaa agccatttcc tctctttttt ctgattgatc 3300 164,326/1 367 cttccctctc acccaggatt agatgctgga aatgaccact tctggagggc agggaacaag 3360 cccttaatct gcataatgag tgttcaataa acagttgtca aactttgaaa 3410 <210> 60 <211> 3336 <212> DNA <213> Homo Sapiens <400> 60 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaaatgagca atccgcggtc 420 tttggaagag gagaaatatg atatgtcagg tgcccgcctg gccctaatac tgtgtgtcac 480 caaagcccgg gaaggttccg aagaagacct ggatgctctg gaacacatgt ttcggcagct 540 gagattcgaa agcaccatga aaagagaccc cactgccgag caattccagg aagagctgga 600 aaaattccag caggccatcg attcccggga agatcccgtc agttgtgcct tcgtggtact 660 catggctcac gggagggaag gcttcctcaa gggagaagat ggggagatgg tcaagctgga 720 gaatctcttc gaggccctga acaacaagaa ctgccaggcc ctgcgagcta agcccaaggt 780 gtacatcata caggcctgtc gaggagaaca aagggacccc ggtgaaacag taggtggaga 840 tgagattgtg atggtcatca aagacagccc acaaaccatc ccaacataca cagatgcctt 900 gcacgtttat tccacggtag agggatacat cgcctaccga catgatcaga aaggctcatg 960 ctttatccag accctggtgg atgtgttcac gaagaggaaa ggacatatct tggaacttct 1020 gacagaggtg acccggcgga tggcagaagc agagctggtt caagaaggaa aagcaaggaa 1080 aacgaaccct gaaatccaaa gcaccctccg gaaacggctg tatctgcagt agaagtagaa 1140 agaccaggag gagctttcct tccagcattc tttctgtctc acagaaattt agaggcagct 1200 cttacctctc cccaagatct tctgttccca aggccaaatg gcacccagtt tcttttccat 1260 cacacccttc atgcaggtcc tcctgtcctt attagagcaa gccagccaaa acttagcaca 1320 aggcatggtg gcaacattaa catcacctcc ctcaggctgg actttctatc tttattaatg 1380 caaccgaaga gacctaagag tgcattcact tatcccactt tctgttcctg tggtcttctt 1440 tctcccatga agcagaaact ggataaagct caagattttc catagacaaa ccaaagccca 1500 ctcatcccct cctaccccaa tccaacctct gctggctcct gcatctcact tggaggtcaa 1560 acctcctcct gaggccaatg cattcccaac ttccagttct ttcctttacc ctggagagtt 1620 agtaaggtaa gaaccattct ttctctccaa aaccactcct ccttggctgg caagttggtg 1680 tcctaactcc gttctcttcc tagctcatgg cctctctaga taataaagtt gtctcctcct 1740 ttctggatct cttcctccta acacccctcc cctgaaaccc tggactctgc cctctctcca 1800 agaaaatcca tctattcaac tattcttgca ttcaattact ctaaatgaga gcgtgttgga 1860 gctatggcaa attccctgtt gtcaccttgc tattttgcag acaacataat atttaacctc 1920 tcataaccag agaggttaaa taatttgtca aatgcaatac agtaagacag aggcaaggac 1980 aaggtttgac ttccagccca gcctcttttc cacaacctgc taaatcctga tccatctgaa 2040 aacttttcta attagtgaag atgactaata aaaattttcc ctatctccaa ggtaggagct 2100 ttctggaagt ttctagaaat tttcaataac caccagccaa ggttacctcc aggtaacctt 2160 gcagcaccag gctggaagtc agatcggctt cactatcttc caactctaca gcctgtatct 2220 ctccatcccc agctttgacc tttcctgctc aagtaaccta cgggcacatc cagcgtcact 2280 aaaaactcag ggcttttctt cccggttact cctccaagcg ttccctggta tcctcaacct 2340 cagatcccag gttcagattt ctgcagtcaa tctatgaccc ctctcttctt gcatccttca 2400' tatgccacca gacaccatgc ccagtccagc ctgattttga aacaactttc atgccggtct 2460 tctcttccct gacatgttac tgtccaggct caagtcctca gcttctcata tctgcatctt 2520 tgcaaccaac ttcctccctt gcctctctgc ttttccatcc cacttttcat gtgtcctcca 2580 taccatctat aacagtgatc tccctggaac actcaagaag acacaacata ccatattatt 2640 taaagaccag ggtactggac agtggctcac acctgtattc ccgactttga gagtctgaag 2700 cgggaggatc acttgaggcc aggagttaag agaccagcct gggcaacaca gcaagaccct 2760 gtctctaaaa aaaaaaatta attaactggg tatggtggca catgcctgta gtcccagcta 2820 ctcaggaggc tgaggtggga ggatgacttg agcccaggag tttgaggctg caaggagcta 2880 tgatcatgcc agtgcatccc agctctaggt gagacagtga gatccggtct ccaaaataaa 2940 tcaatcaatc aaataaagac caaagtcaaa ccgcacatca ggatctctca cacccttcca 3000 attttgccat ctaccagcac ttagctaaac ccatctccca tctcttccac catgaattca 3060 ctctttcaaa aaggctaatg tcttcttact cacccttgcc tctaagcctt tgctatcacc 3120 164,326/1 368 atttccccca agctggaggg ccctccctct ccctttaccc ctcttccact acctcccacc 3180 cctacttttt ccagaaagcc atttcctctc ttttttctga ttgatccttc cctctcaccc 3240' aggattagat gctggaaatg accacttctg gagggcaggg aacaagccct taatctgcat 3300 aatgagtgtt caataaacag ttgtcaaact ttgaaa 3336 <210> 61 <211> 3404 <212> DNA <213> Homo Sapiens <400> 61 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaaatgagca atccgcggtc 420 tttggaagag gagaaatatg atatgtcagg tgcccgcctg gccctaatac tgtgtgtcac 480 caaagcccgg gaaggttccg aagaagacct ggatgctctg gaacacatgt ttcggcagct 540 gagattcgaa agcaccatga aaagagaccc cactgccgag caattccagg aagagctgga 600 aaaattccag caggccatcg attcccggga agatcccgtc agttgtgcct tcgtggtact 660 catggctcac gggagggaag gcttcctcaa gggagaagat ggggagatgg tcaagctgga 720 gaatctcttc gaggccctga acaacaagaa ctgccaggcc ctgcgagcta agcccaaggt 780 gtacatcata caggcctgtc gaggagccac cctgcccagc ccctttcctt acctttctct 840 ctgactttgc ctcctcctct tcttgttgtt tcagaacaaa gggaccccgg tgaaacagta 900 ggtggagatg agattgtgat ggtcatcaaa gacagcccac aaaccatccc aacatacaca 960 gatgccttgc acgtttattc cacggtagag ggatacatcg cctaccgaca tgatcagaaa 1020 ggctcatgct ttatccagac cctggtggat gtgttcacga agaggaaagg acatatcttg 1080 gaacttctga cagaggtgac ccggcggatg gcagaagcag agctggttca agaaggaaaa 1140 gcaaggaaaa cgaaccctga aatccaaagc accctccgga aacggctgta tctgcagtag 1200 aagtagaaag accaggagga gctttccttc cagcattctt tctgtctcac agaaatttag 1260 aggcagctct tacctctccc caagatcttc tgttcccaag gccaaatggc acccagtttc 1320 ttttccatca cacccttcat gcaggtcctc ctgtccttat tagagcaagc cagccaaaac 1380 ttagcacaag gcatggtggc aacattaaca tcacctccct caggctggac tttctatctt 1440 tattaatgca accgaagaga cctaagagtg cattcactta tcccactttc tgttcctgtg 1500 gtcttctttc tcccatgaag cagaaactgg ataaagctca agattttcca tagacaaacc 1560 aaagcccact catcccctcc taccccaatc caacctctgc tggctcctgc atctcacttg 1620 gaggtcaaac ctcctcctga ggccaatgca ttcccaactt ccagttcttt cctttaccct 1680 ggagagttag taaggtaaga accattcttt ctctccaaaa ccactcctcc ttggctggca 1740 agttggtgtc ctaactccgt tctcttccta gctcatggcc tctctagata ataaagttgt 1800 ctcctccttt ctggatctct tcctcctaac acccctcccc tgaaaccctg gactctgccc 1860 tctctccaag aaaatccatc tattcaacta ttcttgcatt caattactct aaatgagagc 1920 gtgttggagc tatggcaaat tccctgttgt caccttgcta ttttgcagac aacataatat 1980 ttaacctctc ataaccagag aggttaaata atttgtcaaa tgcaata.cag taagacagag 2040 gcaaggacaa ggtttgactt ccagcccagc ctcttttcca caacctgcta aatcctgatc 2100 catctgaaaa cttttctaat tagtgaagat gactaataaa aattttccct atctccaagg 2160 taggagcttt ctggaagttt ctagaaattt tcaataacca ccagccaagg ttacctccag 2220 gtaaccttgc agcaccaggc tggaagtcag atcggcttca ctatcttcca actctacagc 2280 ctgtatctct ccatccccag ctttgacctt tcctgctcaa gtaacctacg ggcacatcca 2340 gcgtcactaa aaactcaggg cttttcttcc cggttactcc tccaagcgtt ccctggtatc 2400 ctcaacctca gatcccaggt tcagatttct gcagtcaatc tatgacccct ctcttcttgc 2460 atccttcata tgccaccaga caccatgccc agtccagcct gattttgaaa caactttcat 2520 gccggtcttc tcttccctga catgttactg tccaggctca agtcctcagc ttctcatatc 2580 tgcatctttg caaccaactt ' cctcccttgc ctctctgctt ttccatccca cttttcatgt 2640 gtcctccata ccatctataa cagtgatctc cctggaacac tcaagaagac acaacatacc 2700 atattattta aagaccaggg tactggacag tggctcacac ctgtattccc gactttgaga 2760 gtctgaagcg ggaggatcac ttgaggccag gagttaagag accagcctgg gcaacacagc 2820 aagaccctgt ctctaaaaaa aaaaattaat taactgggta tggtggcaca tgcctgtagt 2880 cccagctact caggaggctg aggtgggagg atgacttgag cccaggagtt tgaggctgca 2940 aggagctatg atcatgccag tgcatcccag ctctaggtga gacagtgaga tccggtctcc 3000 164,326/1 369 aaaataaatc aatcaatcaa ataaagacca aagtcaaacc gcacatcagg atctctcaca 3060 cccttccaat tttgccatct accagcactt agctaaaccc atctcccatc tcttccacca 3120 tgaattcact ctttcaaaaa ggctaatgtc ttcttactca cccttgcctc taagcctttg 3180 ctatcaccat ttcccccaag ctggagggcc ctccctctcc ctttacccct cttccactac 3240 ctcccacccc tactttttcc agaaagccat ttcctctctt ttttctgatt gatccttccc 3300 tctcacccag gattagatgc tggaaatgac cacttctgga gggcagggaa caagccctta 3360 atctgcataa tgagtgttca ataaacagtt gtcaaacttt gaaa 3404 <210> 62 <211> 3336 <212> DNA <213> Homo Sapiens <400> 62 ctgactcatt tagactctct gcctaggcca cctttgccag agggagtccc ctcagccttg 60 cgatcactca tcccattggc gttggctcca tttccacacc acagctgtgt gccaagggtg 120 tgtcatgagg tttcttgagt gacagaaaac tcaccgacaa taaagggcca ggtgattgtg 180 ccacccgatt catagaccag gcttctcagg agaaaccccg ggagattcca cactgtcagc 240 cccttctcca agatcagtac gtgggcctga ctcctcctcg gtgcccagct cagtattggc 300 aactaggaga gtagtgagat tgaacttggc cttgaggaac agctgcctct agagttggat 360 cagacaaggg tgctgagagc cgggactcac aaccaaagga gaaatgagca atccgcggtc 420 tttggaagag gagaaatatg atatgtcagg tgcccgcctg gccctaatac tgtgtgtcac 480 caaagcccgg gaaggttccg aagaagacct ggatgctctg gaacacatgt ttcggcagct 540 gagattcgaa agcaccatga aaagagaccc cactgccgag caattccagg aagagctgga 600 aaaattccag caggccatcg attcccggga agatcccgtc agttgtgcct tcgtggtact 660 catggctcac gggagggaag gcttcctcaa gggagaagat ggggagatgg tcaagctgga 720 gaatctcttc gaggccctga acaacaagaa ctgccaggcc ctgcgagcta agcccaaggt 780 gtacatcata caggcctgtc gaggagaaca aagggacccc ggtgaaacag taggtggaga 840 tgagattgtg atggtcatca aagacagccc acaaaccatc ccaacataca cagatgcctt 900 gcacgtttat tccacggtag agggatacat cgcctaccga catgatcaga aaggctcatg 960 ctttatccag accctggtgg atgtgttcac gaagaggaaa ggacatatct tggaacttct 1020 gacagaggtg acccggcgga tggcagaagc agagctggtt caagaaggaa aagcaaggaa 1080 aacgaaccct gaaatccaaa gcaccctccg gaaacggctg tatctgcagt agaagtagaa 1140 agaccaggag gagctttcct tccagcattc tttctgtctc acagaaattt agaggcagct 1200 cttacctctc cccaagatct tctgttccca aggccaaatg gcacccagtt tcttttccat 1260 cacacccttc atgcaggtcc tcctgtcctt attagagcaa gccagccaaa acttagcaca 1320 aggcatggtg gcaacattaa catcacctcc ctcaggctgg actttctatc tttattaatg 1380 caaccgaaga gacctaagag tgcattcact tatcccactt tctgttcctg tggtcttctt 1440 tctcccatga agcagaaact ggataaagct caagattttc catagacaaa ccaaagccca 1500 ctcatcccct cctaccccaa tccaacctct gctggctcct gcatctcact tggaggtcaa 1560 acctcctcct gaggccaatg cattcccaac ttccagttct ttcctttacc ctggagagtt 1620 agtaaggtaa gaaccattct ttctctccaa aaccactcct ccttggctgg caagttggtg 1680 tcctaactcc gttctcttcc tagctcatgg cctctctaga taataaagtt gtctcctcct 1740 ttctggatct cttcctccta acacccctcc cctgaaaccc tggactctgc cctctctcca 1800 agaaaatcca tctattcaac tattcttgca ttcaattact ctaaatgaga gcgtgttgga 1860 gctatggcaa attccctgtt gtcaccttgc tattttgcag acaacataat atttaacctc 1920 tcataaccag agaggttaaa taatttgtca aatgcaatac agtaagacag aggcaaggac 1980 aaggtttgac ttccagccca gcctcttttc cacaacctgc taaatcctga tccatctgaa 2040 aacttttcta attagtgaag atgactaata aaaattttcc ctatctccaa ggtaggagct 2100 ttctggaagt ttctagaaat tttcaataac caccagccaa ggttacctcc aggtaacctt 2160 gcagcaccag gctggaagtc agatcggctt cactatcttc caactctaca gcctgtatct 2220 ctccatcccc agctttgacc tttcctgctc aagtaaccta cgggcacatc cagcgtcact 2280 aaaaactcag ggcttttctt cccggttact cctccaagcg ttccctggta tcctcaacct 2340 cagatcccag gttcagattt ctgcagtcaa tctatgaccc ctctcttctt gcatccttca 2400 tatgccacca gacaccatgc ccagtccagc ctgattttga aacaactttc atgccggtct 2460 tctcttccct gacatgttac tgtccaggct caagtcctca gcttctcata tctgcatctt 2520 tgcaaccaac ttcctccctt gcctctctgc ttttccatcc cacttttcat gtgtcctcca 2580 taccatctat aacagtgatc tccctggaac actcaagaag acacaacata ccatattatt 2640 taaagaccag ggtactggac agtggctcac acctgtattc ccgactttga gagtctgaag 2700 cgggaggatc acttgaggcc aggagttaag agaccagcct gggcaacaca gcaagaccct 2760 gtctctaaaa aaaaaaatta attaactggg tatggtggca catgcctgta gtcccagcta 2820 164,326/1 370 ctcaggaggc tgaggtggga ggatgacttg agcccaggag tttgaggctg caaggagcta 2880 tgatcatgcc agtgcatccc agctctaggt gagacagtga gatccggtct ccaaaataaa 2940 tcaatcaatc aaataaagac caaagtcaaa ccgcacatca ggatctctca cacccttcca 3000 attttgccat ctaccagcac ttagctaaac ccatctccca tctcttccac catgaattca 3060 ctctttcaaa aaggctaatg tcttcttact cacccttgcc tctaagcctt tgctatcacc 3120 atttccccca agctggaggg ccctccctct ccctttaccc ctcttccact acctcccacc 3180 cctacttttt ccagaaagcc atttcctctc ttttttctga ttgatccttc cctctcaccc 3240 aggattagat gctggaaatg accacttctg gagggcaggg aacaagccct taatctgcat 3300 aatgagtgtt caataaacag ttgtcaaact ttgaaa 3336 <210> 63 <211> 966 <212> DNA <213> Homo Sapiens <400> 63 atggggaaat gccaagagta tgacaaaagt ctgtctgtgc agccagagaa gagaacagga 60 ctcagagatg agaatggaga atgtggacag acattcagac tcaaggaaga gcaagggagg 120 gctttcaggg gaagttcagt ccaccagaag ctggtgaatg acccacggga gacacaggaa 180 gtttttgggg gcggagtggg ggacattgtg ggacgggatc tcagtattag cttcagaaac 240 tctgagacct ctgcaagtga ggaggagaaa tatgatatgt caggtgcccg cctggcccta 300 atactgtgtg tcaccaaagc ccgggaaggt tccgaagaag acctggatgc tctggaacac 360 atgtttcggc agctgagatt cgaaagcacc atgaaaagag accccactgc cgagcaattc 420 caggaagagc tggaaaaatt ccagcaggcc atcgattccc gggaagatcc cgtcagttgt 480 gccttcgtgg tactcatggc tcacgggagg gaaggcttcc tcaagggaga agatggggag 540 atggtcaagc tggagaatct cttcgaggcc ctgaacaaca agaactgcca ggccctgcga 600 gctaagccca aggtgtacat catacaggcc tgtcgaggag aacaaaggga ccccggtgaa 660 acagtaggtg gagatgagat tgtgatggtc atcaaagaca gcccacaaac catcccaaca 720 tacacagatg ccttgcacgt ttattccacg gtagagggat acatcgccta ccgacatgat 780 cagaaaggct catgctttat ccagaccctg gtggatgtgt tcacgaagag gaaaggacat 840 atcttggaac ttctgacaga ggtgacccgg cggatggcag aagcagagct ggttcaagaa 900 ggaaaagcaa ggaaaacgaa ccctgaaatc caaagcaccc tccggaaacg gctgtatctg 960 cagtag 966 <210> 64 <211> 230 <212> PRT <213> Homo Sapiens <400> 64 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Pro Thr 165 170 175 Pro Phe Gin Asp Pro Leu Tyr Leu Pro Ser Glu Ala Pro Pro Asn Pro 180 185 190 Pro Leu Trp As Ser Gin Asp Thr Ser Pro Thr Asp Met He Arg Lys 195 200 205 Ala His Ala Leu Ser Arg Pro Trp Trp Met Cys Ser Arg Arg Gly Lys 210 215 220 Asp lie Ser Trp Asn Phe 225 230 <210> 65 <211> 146 <212> PRT <213> Homo Sapiens <400> 65 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Ala Thr Leu Pro Ser Pro Phe Pro Tyr Leu 130 135 140 Ser Leu 145 <210> 66 <211> 321 <212> PRT <213> Homo : Bapiens <400> 66 Met Gly Lys Cys Gin Glu Tyr Asp Lys Ser Leu Ser Val Gin Pro Glu 1 5 10 15 Lys Arg Thr Gly Leu Arg Asp Glu Asn Gly Glu Cys Gly Gin Thr Phe -25 30 Arg Leu Lys Glu Glu Gin Gly Arg Ala Phe Arg Gly Ser Ser Val His 40 45 Gin Lys Leu Val Asn Asp Pro Arg Glu Thr Gin Glu Val Phe Gly Gly 50 55 60 Gly Val Gly Asp He Val Gly Arg Asp Leu Ser He Ser Phe Arg Asn 65 70 75 80 Ser Glu Thr Ser Ala Ser Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala 85 90 95 Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser Glu 100 105 110 Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu 115 120 125 Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu 130 135 140 Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser Cys 145 150 155 160 Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys Gly 165 170 175 Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu Asn 180 185 190 Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He He 195 200 205 Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly Gly 210 215 220 Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro Thr 225 230 235 240 Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He Ala 245 250 255 Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val Asp 260 265 270 Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu Val 275 280 285 Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala Arg 290 295 300 Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr Leu 305 310 315 320 Gin <210> 67 <211> 242 <212> PRT <213> Homo Sapiens <400> 67 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala lie Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu lie Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 68 <211> 230 <212> PRT <213> Homo Sapiens <400> 68 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Pro Thr 165 170 175 Pro Phe Gin Asp Pro Leu Tyr Leu Pro Ser Glu Ala Pro Pro Asn Pro 180 185 190 Pro Leu Trp Asn Ser Gin Asp Thr Ser Pro Thr Asp Met He Arg Lys 195 200 205 Ala His Ala Leu Ser Arg Pro Trp Trp Met Cys Ser Arg Arg Gly Lys 210 215 220 Asp He Ser Trp Asn Phe 225 230 <210> 69 <211> 242 <212> PRT <213> Homo Sapiens <400> 69 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 70 <211> 146 <212> PRT <213> Homo Sapiens <400> 70 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Ala Thr Leu Pro Ser Pro Phe Pro Tyr Leu 130 135 140 Ser Leu 145 <210> 71 <211> 242 <212> PRT <213> Homo Sapiens <400> 71 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 72 <211> 321 <212> PRT <213> Homo Sapiens <400> 72 Met Gly Lys Cys Gin Glu Tyr Asp Lys Ser Leu Ser Val Gin Pro Glu 1 5 10 15 Lys Arg Thr Gly Leu Arg Asp Glu Asn Gly Glu Cys Gly Gin Thr Phe 25 30 Arg Leu Lys Glu Glu Gin Gly Arg Ala Phe Arg Gly Ser Ser Val His 40 45 Gin Lys Leu Val Asn Asp Pro Arg Glu Thr Gin Glu Val Phe Gly Gly 50 55 60 Gly Val Gly Asp He Val Gly Arg Asp Leu Ser He Ser Phe Arg Asn 65 70 75 80 Ser Glu Thr Ser Ala Ser Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala 85 90 95 Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser Glu 100 105 110 Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe Glu 115 120 125 Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu Leu 130 135 140 Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser Cys 145 150 155 160 Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys Gly 165 170 175 Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu Asn 180 185 190 Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He He 195 200 205 Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly Gly 210 215 220 Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro Thr 225 230 235 240 Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He Ala 245 250 255 Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val Asp 260 265 270 Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu Val 275 280 285 Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala Arg 290 295 300 Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr Leu 305 310 315 320 Gin <210> 73 <211> 242 <212> PRT <213> Homo Sapiens <400> 73 Met Ser Asn Pro Arg Ser Leu Glu Glu Glu Lys Tyr Asp Met Ser Gly 1 5 10 15 Ala Arg Leu Ala Leu He Leu Cys Val Thr Lys Ala Arg Glu Gly Ser 25 30 Glu Glu Asp Leu Asp Ala Leu Glu His Met Phe Arg Gin Leu Arg Phe 40 45 Glu Ser Thr Met Lys Arg Asp Pro Thr Ala Glu Gin Phe Gin Glu Glu 50 55 60 Leu Glu Lys Phe Gin Gin Ala He Asp Ser Arg Glu Asp Pro Val Ser 65 70 75 80 Cys Ala Phe Val Val Leu Met Ala His Gly Arg Glu Gly Phe Leu Lys 85 90 95 Gly Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Leu 100 105 110 Asn Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val Tyr He 115 120 125 He Gin Ala Cys Arg Gly Glu Gin Arg Asp Pro Gly Glu Thr Val Gly 130 135 140 Gly Asp Glu He Val Met Val He Lys Asp Ser Pro Gin Thr He Pro 145 150 155 160 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Tyr He 165 170 175 Ala Tyr Arg His Asp Gin Lys Gly Ser Cys Phe He Gin Thr Leu Val 180 185 190 Asp Val Phe Thr Lys Arg Lys Gly His He Leu Glu Leu Leu Thr Glu 195 200 205 Val Thr Arg Arg Met Ala Glu Ala Glu Leu Val Gin Glu Gly Lys Ala 210 215 220 Arg Lys Thr Asn Pro Glu He Gin Ser Thr Leu Arg Lys Arg Leu Tyr 225 230 235 240 Leu Gin <210> 74 <211> 64 <212> PRT <213> Homo Sapiens <400> 74 His Val Tyr Ser Thr Val Glu Gly Pro Thr Pro Phe Gin Asp Pro Leu 1 5 10 15 Tyr Leu Pro Ser Glu Ala Pro Pro Asn Pro Pro Leu Trp Asn Ser Gin 25 30 Asp Thr Ser Pro Thr Asp Met He Arg Lys Ala His Ala Leu Ser Arg 40 45 Pro Trp Trp Met Cys Ser Arg Arg Gly Lys Asp He Ser Trp Asn Phe 50 55 60 <210> 75 <211> 65 <212> PRT <213> Homo Sapiens <400> 75 Leu His Val Tyr Ser Thr Val Glu Gly Pro Thr Pro Phe Gin Asp Pro 1 5 10 15 Leu Tyr Leu Pro Ser Glu Ala Pro Pro Asn Pro Pro Leu Trp Asn Ser 25 30 Gin Asp Thr Ser Pro Thr Asp Met He Arg Lys Ala His Ala Leu Ser 40 45 Arg Pro Trp Trp Met Cys Ser Arg Arg Gly Lys Asp He Ser Trp Asn 50 55 60 Phe 65 <210> 76 <211> 70 <212> PRT <213> Homo Sapiens <400> 76 Thr Tyr Thr Asp Ala Leu His Val Tyr Ser Thr Val Glu Gly Pro Thr 1 5 10 15 Pro Phe Gin Asp Pro Leu Tyr Leu Pro Ser Glu Ala Pro Pro Asn Pro 25 30 Pro Leu Trp Asn Ser Gin Asp Thr Ser Pro Thr Asp Met He Arg Lys 40 45 Ala His Ala Leu Ser Arg Pro Trp Trp Met Cys Ser Arg Arg Gly Lys 50 55 60 Asp He Ser Trp Asn Phe 65 70 <210> 77 <211> 20 <212> PRT <213> Homo Sapiens <400> 77 Tyr He He Gin Ala Cys Arg Gly Ala Thr Leu Pro Ser Pro Phe Pro 1 5 10 15 Tyr Leu Ser Leu <210> 78 <211> 21 <212> PRT <213> Homo Sapiens <400> 78 Val Tyr He He Gin Ala Cys Arg Gly Ala Thr Leu Pro Ser Pro Phe 1 5 10 15 Pro Tyr Leu Ser Leu <210> 79 <211> 26 <212> PRT <213> Homo Sapiens <400> 79 Arg Ala Lys Pro Lys Val Tyr He He Gin Ala Cys Arg Gly Ala Thr 1 5 10 15 Leu Pro Ser Pro Phe Pro Tyr Leu Ser Leu 25 <210> 80 <211> 94 <212> PRT <213> Homo Sapiens <400> 80 Met Gly Lys Cys Gin Glu Tyr Asp Lys Ser Leu Ser Val Gin Pro Glu 1 5 10 15 Lys Arg Thr Gly Leu Arg Asp Glu Asn Gly Glu Cys Gly Gin Thr Phe . 25 30 Arg Leu Lys Glu Glu Gin Gly Arg Ala Phe Arg Gly Ser Ser Val His 40 45 Gin Lys Leu Val Asn Asp Pro Arg Glu Thr Gin Glu Val Phe Gly Gly 50 55 60 Gly Val Gly Asp He Val Gly Arg Asp Leu Ser He Ser Phe Arg Asn 65 70 75 80 Ser Glu Thr Ser Ala Ser Glu. Glu Glu Lys Tyr Asp Met Ser 85 90 <210> 81 <211> 95 <212> PRT <213> Homo Sapiens <400> 81 Met Gly Lys Cys Gin Glu Tyr Asp Lys Ser Leu Ser Val Gin Pro Glu 1 5 10 15 Lys Arg Thr Gly Leu Arg Asp Glu Asn Gly Glu Cys Gly Gin Thr Phe 25 30 Arg Leu Lys Glu Glu Gin Gly Arg Ala Phe Arg Gly Ser Ser Val His 40 45 Gin Lys Leu Val Asn Asp Pro Arg Glu Thr Gin Glu Val Phe Gly Gly 50 55 60 Gly Val Gly Asp He Val Gly Arg Asp Leu Ser He Ser Phe Arg Asn 65 70 75 80 Ser Glu Thr Ser Ala Ser Glu Glu Glu Lys Tyr Asp Met Ser Gly 85 90 95 <210> 82 <211> 100 <212> PRT <213> Homo Sapiens <400> 82 Met Gly Lys Cys Gin Glu Tyr Asp Lys Ser Leu Ser Val Gin Pro Glu 1 5 10 15 Lys Arg Thr Gly Leu Arg Asp Glu Asn Gly Glu Cys Gly Gin Thr Phe 25 30 Arg Leu Lys Glu Glu Gin Gly Arg Ala Phe Arg Gly Ser Ser Val His 40 45 Gin Lys Leu Val Asn Asp Pro Arg Glu Thr Gin Glu Val Phe Gly Gly 50 55 60 Gly Val Gly Asp He Val Gly Arg Asp Leu Ser He Ser Phe Arg Asn 65 70 75 80 Ser Glu Thr Ser Ala Ser Glu Glu Glu Lys Tyr Asp Met Ser Gly Ala 85 90 95 Arg Leu Ala Leu 100 <210> 83 <211> 17 <212> PRT <213> Homo Sapiens <400> 83 Gly Ala Arg Leu Ala Leu He Leu Arg Val Thr Lys Ala Arg Glu Gly 1 5 10 15 Ser <210> 84 <211> 19 <212> PRT <213> Homo Sapiens <400> 84 Ser Gly Ala Arg Leu Ala Leu He Leu Arg Val Thr Lys Ala Arg Glu 1 5 10 15 Gly Ser Glu <210> 85 <211> 29 <212> PRT <213> Homo Sapiens <400> 85 Glu Lys Tyr Asp Met Ser Gly Ala Arg Leu Ala Leu He Leu Arg Val 1 5 10 15 Thr Lys Ala Arg Glu Gly Ser Glu Glu Asp Leu Asp Ala 25 <210> 86 <211> 17 <212> PRT <213> Homo Sapiens <400> 86 Lys Leu Glu Asn Leu Phe Glu Ala Met Asn Asn Lys Asn Cys Gin Ala 1 5 10 15 Leu <210> 87 <211> 19 <212> PRT <213> Homo Sapiens <400> 87 Val Lys Leu Glu Asn Leu Phe Glu Ala Met Asn Asn Lys Asn Cys Gin 1 5 10 15 Ala Leu Arg <210> 88 <211> 29 <212> PRT <213> Homo Sapiens <400> 88 Glu Asp Gly Glu Met Val Lys Leu Glu Asn Leu Phe Glu Ala Met Asn 1 5 10 15 Asn Lys Asn Cys Gin Ala Leu Arg Ala Lys Pro Lys Val 25

Claims (20)

381 164,326/2 What is Claimed is:

1. An isolated polynucleotide that encodes a protein consisting of the polypeptide sequence shown in SEQ ID NO: 18, 19, 20, 21, 22, or 23.

2. The polynucleotide of claim 1, wherein the polynucleotide is selected from the group consisting of: (a) a polynucleotide comprising the sequence of SEQ ID NO:2, from nucleotide residue numbers 404 through 1132; (b) a polynucleotide comprising the sequence of SEQ ID NO:4, from nucleotide residue numbers 404 through 1096; (c) a polynucleotide comprising the sequence of SEQ ID NO:6, from nucleotide residue numbers 404 through 844; (d) a polynucleotide comprising the sequence of SEQ ID NO:8, from nucleotide residue numbers 1 through 966; (e) a polynucleotide comprising the sequence of SEQ ID NO: 10, from nucleotide residue numbers 404 through 1132; (f) a polynucleotide comprising the sequence of SEQ ID NO:2, from nucleotide residue numbers 404 through 1 132, wherein the nucleotide residue at 2027 is T; (g) a polynucleotide comprising the sequence of SEQ ID NO:2, from nucleotide residue numbers 404 through 1132, wherein the nucleotide residue at 2037 is C; (h) a polynucleotide comprising the sequence of SEQ ID NO:2, from nucleotide residue numbers 404 through 1 132, wherein the nucleotide residue at 2268 is G; and (i) a polynucleotide comprising the sequence of SEQ ID NO:2, from nucleotide residue numbers 404 through 1 132, wherein the nucleotide residue at 3196 is T.

3. A recombinant expression vector comprising a polynucleotide of any one of claims 1-2, wherein the vector is a viral vector.

4. The recombinant expression vector of claim 3, wherein the viral vector is selected from the group consisting of vaccinia, fowlpox, canarypox, adenovirus, influenza, poliovirus, adeno-associated virus, lentivirus, and Sindbis virus.

5. A host cell that contains an expression vector of claim 3. 382 164,326/2

6. A process for producing a protein comprising culturing the host cell of claim 5 under conditions sufficient for the production of the protein, wherein the amino acid sequence of the protein is selected from the group consisting of SEQ ID NO: 18, 19, 20, 21, 22, and 23.

7. The process of claim 6, further comprising recovering the protein so produced.

8. The process of claim 7, wherein the protein is recovered using chromatography.

9. A composition comprising a pharmaceutically acceptable carrier and the viral vector of claims 3 or 4.

10. The composition of claim 9, wherein the viral vector is fowlpox.

11. 1 1. An in vitro method for detecting the presence of a protein comprising the amino acid sequence of SEQ ID NO: 18, 19, 20, 21, 22, or 23 or a polynucleotide comprising the polynucleotide sequence of SEQ ID NO: 4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, or SEQ ID NO:2, wherein 2027 is C or T, 2037 is T or C, 2268 is A or G, and 3196 is A or T, in a test sample comprising: contacting the sample with an antibody or polynucleotide, respectively, that specifically binds to the protein or polynucleotide, respectively; and detecting binding of protein or polynucleotide, respectively, in the sample thereto.

12. The method of claim 1 1, wherein the polynucleotide is an mRNA.

13. The method of claim 11, wherein the polynucleotide is a cDNA produced from the sample by reverse transcription.

14. The method of any one of claims 11-13, further comprising comparing an amount of binding of the antibody or polynucleotide that specifically binds to the protein or the polynucleotide in the test sample to the presence of the protein or polynucleotide in a corresponding normal sample.

15. The method of claim 14, wherein the presence of elevated polynucleotide or protein in the test sample relative to the normal tissue sample provides an indication of the presence of cancer. 383 164,326/2

16. The method of claim 15, wherein the cancer is selected from the group consisting of cancer of the prostate, bladder, and breast.

17. Use of a protein comprising the amino acid sequence of SEQ ID NO: 18, 19, 20, 21, 22, or 23, for the preparation of a medicament to induce an immune response from a subject, whereby a T cell or B cell is activated.

18. The use of claim 17, wherein the activated B cell generates antibodies that specifically bind to the protein.

19. The use of claim 17, wherein the activated T cell is a cytotoxic T cell (CTL), whereby the activated CTL kills an autologous cell that expresses the protein.

20. The use of claim 17, wherein the activated T cell is a helper T cell (HTL), whereby the activated HTL secretes cytokines that facilitate the cytotoxic activity of a CTL or the antibody producing activity of a B cell. For the Applicant WOLFF, BREGl D GOLLER