AU2002318112A1 - Nucleic acids and corresponding proteins useful in the detection and treatment of various cancers - Google Patents

Description

NUCLEIC ACIDS AND CORRESPONDING PROTEINS USEFUL IN THE DETECTION AND TREATMENT OF VARIOUS CANCERS

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to United States Provisional Application Serial No. 60/283,112 filed 10 April 2001; United States Provisional Application Serial No. 60/282,739, filed 10 April 2001; and, United States Provisional Application Serial No. 60/286,630 filed 25 April 2001. The content of each of which is hereby incorporated by reference herein in its entirety.

STATEMENT OF RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH

Not applicable.

FIELD OF THE INVENTION

The invention described herein relates to genes and their respective encoded proteins, such as those set forth, e.g., in Figure 2, expressed in certain cancers, and to diagnostic and therapeutic methods and compositions useful in the management of cancers that express a gene of Figure 2.

BACKGROUND OF THE INVENTION

Cancer is the second leading cause of human death next to coronary disease. Worldwide, millions of people die from cancer every year. In the United States alone, as reported by the American Cancer Society, cancer causes the death of well over a half-million people annually, with over 1.2 million, new cases diagnosed per year. While deaths from heart disease have been declining significantly, those resulting from cancer generally are on the rise. In the early part ofthe next century, cancer is predicted to become the leading cause of death.

Worldwide, several cancers stand out as the leading killers. In particular, carcinomas ofthe 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 ofthe 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 has been a very useful tool, however its specificity and 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 ofthe 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 exhibited the capacity to mimic the transition from androgen dependence to androgen independence (Klein et al, 1997, Nat. Med. 3:402). More recently identifiέd prostate cancer markers include PCTA-1 (Su et al, 1996, Proc. Natl. Acad. Sci. USA 93: 7252), prostate-specific membrane (PSM) antigen (Pinto et al, Clin Cancer Res 1996 Sep 2 (9): 1445-51), STEAP (Hubert, et al, Proc Natl Acad Sci U S A. 1999 Dec 7; 96(25): 14523-8) and prostate stem cell antigen (PSCA) (Reiter et 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 further improve diagnosis and therapy.

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 ofthe renal pelvis or ureter. The incidence of renal cell adenocarcinoma is estimated at more than 29,000 cases in the United States, and more than 11,600 patients died of this disease in 1998. Transitional cell carcinoma is less frequent, with an incidence of approximately 500 cases per year in the United States.

Surgery has been the primary therapy for renal cell 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 aggressively in appropriate patients with a possibility of durable responses. Nevertheless, there is a remaining need for effective therapies for these patients.

Of all new cases of cancer in the United States, bladder cancer represents approximately 5 percent in men (fifth most common neoplasm) and 3 percent in women (eighth most common neoplasm). The incidence is increasing slowly, concurrent with an increasing older population. In 1998, there was an estimated 54,500 cases, including 39,500 in men and 15,000 in women. The age-adjusted incidence in the United States is 32 per 100,000 for men and 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 an estimated 11,000 deaths from bladder cancer in 1998 (7,800 in men and 3,900 in women). Bladder cancer incidence and mortality strongly increase with age and will be an increasing problem as the population becomes more elderly.

Most bladder cancers recur in the bladder. Bladder cancer is managed with a combination of transurethral resection ofthe bladder (TUR) and intravesical chemotherapy or immunotherapy. The multifocal and recurrent nature of bladder cancer points out the 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 the United States, including 93,800 cases of colon cancer and 36,400 of rectal cancer. Colorectal cancers are the 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 deaths (47,700 from colon cancer, 8,600 from rectal cancer) in 2000, accounting for about 11% of all U.S. cancer deaths.

At present, surgery is the 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 the bowel wall or has spread to the 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 the 1990s, the rate of increase among women began to slow. In 1996, the incidence rate in women was 42.3 per 100,000.

Lung and bronchial cancer caused an estimated 156,900 deaths in 2000, accounting for 28% of all cancer deaths. During 1992-1996, mortality from lung cancer declined significantly among men (-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 than breast cancer, which, for over 40 years, was the major cause of cancer death in women. Decreasing lung cancer incidence and mortality rates most likely resulted from decreased smoking rates over the previous 30 years; however, decreasing smoking patterns among women lag behind those of men. Of concern, although the declines in adult tobacco use have slowed, tobacco use in youth is increasing again.

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

An estimated 182,800 new invasive cases of breast cancer were expected to occur among women in the United States during 2000. Additionally, about 1,400 new cases of breast cancer were expected to be diagnosed in men in 2000. After increasing about 4% per year in the 1980s, breast cancer incidence rates in women have leveled off in the 1990s to about 110.6 cases per 100,000.

In the U.S. alone, there were an estimated 41,200 deaths (40,800 women, 400 men) in 2000 due to breast cancer. Breast cancer ranks second among cancer deaths in women. According to the most recent data, mortality rates declined significantly during 1992-1996 with the largest decreases in younger women, both white and black. These decreases were probably the result of earlier detection and improved treatment.

Taking into account the medical circumstances and the patient's preferences, treatment of breast cancer may involve lumpectomy (local removal ofthe tumor) and removal ofthe lymph nodes under the arm; mastectomy (surgical removal ofthe breast) and removal ofthe 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 ofthe 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 ofthe female reproductive system.

Surgery, radiation therapy, and chemotherapy are treatment options for ovarian cancer. Surgery usually includes the removal of one or both ovaries, the 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 intra-abdominal 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.

SUMMARY OF THE INVENTION

The present invention relates to genes and respective encoded proteins set forth in Figure 2, that have now been found to be over-expressed in the cancer(s) listed in Table I. Northern blot expression analysis ofthe genes of Figure 2 in normal tissues shows a restricted expression pattern in adult tissues. The nucleotide (Figure 2) and amino acid (Figure 2, and Figure 3) sequences of Figure 2 are provided. The tissue-related expression profile ofthe genes set forth in Figure 2 in normal adult tissues, combined with the over-expression observed in the tumors listed in Table I, shows that the genes of Figure 2 are aberrantly over-expressed in certain cancers, and thus serves as a useful diagnostic, prophylactic, prognostic, and/or therapeutic target for cancers ofthe tissue(s) such as those listed in Table I.

The invention provides polynucleotides corresponding or complementary to all or part ofthe genes of Figure 2, corresponding/related mRNAs, coding and/or complementary sequences, preferably in isolated form, including polynucleotides encoding Figure 2-related proteins and fragments of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or more than 25 contiguous amino acids of a Figure 2-related protein; at least 30, 35, 40, 45, 50, 55, 60, 65, 70, 80, 85, 90, 95, 100 or more than 100 contiguous amino acids of a Figure 2-related protein, as well as the peptides/proteins themselves; DNA, RNA, DNA/RNA hybrids, and related molecules such as, polynucleotides or oligonucleotides complementary or having at least a 90% homology to the genes set forth in Figure 2 or mRNA sequences or parts thereof, and polynucleotides or oligonucleotides that hybridize to the genes set forth in Figure 2, mRNAs, or to polynucleotides that encode proteins of Figure 2 or figure 3 or analogs or variants thereof; or to polynucleotides that encode proteins of fragments of a peptide of Figure 2 or Figure 3 such as set forth in Tables V to XVIII, Table XX, Tables XXIII to XXVI, or analogs or variants thereof; or to polynucleotides that encode fragments/subsequences of a peptide of Figure 2 or Figure 3 such as any 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, 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, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 505, 510, 515, 520, 525, 530, 535, 540, 545, 550, 555, 560, 565, 570, 575, 580, 585, 590, 595, 600, 605, 610, 615, 620, 625, 630, 635, 640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690, 695, 700, 705, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760, 765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 815, 820, 825, 830, 835, 840, 845, 850, 855, 860, 865, 870, 875, 880, 885, 890, 895, 900, 905, 910, 915, 920, 925, 930, 935, 940, 945, 950, 955, 960, 965, 970, 975, 980, 985, 990, 995, 1000, 1025, 1050, 1075, 1100, 1125, 1150, 1175, 1200, etc., or more contiguous amino acids of a peptide of Figure 2 or 3, or an analog or variant thereof.

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

The invention further provides methods for detecting the presence and status of Figure 2 polynucleotides and proteins in various biological samples, as well as methods for identifying cells that express the genes set forth in Figure 2. A typical embodiment of this invention provides methods for monitoring the Figure 2 gene products in a tissue or hematology sample having or suspected of having some form of growth dysregulation such as cancer.

The invention further provides various immunogenic or therapeutic compositions and strategies for treating cancers that express a gene set forth in Figure 2 such as cancers of tissues listed in Table I, including therapies aimed at inhibiting the transcription, translation, processing or function ofthe genes of Figure 2 as well as cancer vaccines. In one aspect, the invention provides compositions, and methods comprising them, for treating a cancer that expresses a gene set forth in Figure 2 in a human subject wherein the composition comprises a carrier suitable for human use and a human unit dose of one or more than one agent that inhibits the production or function of a gene or proteins of Figure 2. Preferably, the carrier is a uniquely for use in humans. In another aspect ofthe invention, the agent is a moiety that is immunoreactive with a protein of Figure 2. Non-limiting examples of such moieties include, but are not limited to, antibodies (such as single chain, monoclonal, polyclonal, humanized, chimeric, or human antibodies), functional equivalents thereof (whether naturally occurring or synthetic), and combinations thereof. The antibodies can be conjugated to a diagnostic or therapeutic moiety. In another aspect, the agent is a small molecule as defined herein.

In another aspect, the agent comprises one or more than one peptide which comprises a cytotoxic T lymphocyte (CTL) epitope that binds an HLA class I molecule in a human to elicit a CTL response to a protein of Figure 2 and/or one or more than one peptide which comprises a helper T lymphocyte (HTL) epitope which binds an HLA class II molecule in a human to elicit an HTL response. The peptides ofthe invention may be on the same or on one or more separate polypeptide molecules. In a further aspect ofthe invention, the agent comprises one or more than one nucleic acid molecule that expresses one or more than one ofthe CTL or HTL response stimulating peptides as described above. In yet another aspect ofthe invention, the one or more than one nucleic acid molecule may express a moiety that is immunologically reactive with a protein of Figure 2 as described above. The one or more than one nucleic acid molecule may also be, or encodes, a molecule that inhibits production of a protein set forth in Figure 2. Non-limiting examples of such molecules include, but are not limited to, those complementary to a nucleotide sequence essential for production of a protein of Figure 2 (e.g. antisense sequences or molecules that form a triple helix with a nucleotide double helix essential for production of a protein set forth in Figure 2) or a ribozyme effective to lyse mRNA (sense or antisense) encoded by a gene of Figure 2.

Please note, to determine the starting position of any peptide set forth in Tables V-XVIII and Tables XXIII to XXVI (collectively HLA Peptide Tables) respective to its parental protein in Figure 2 or Figure 3, reference is made to its respective protein.

One embodiment ofthe invention comprises an HLA peptide, that occurs at least twice in Tables V-XVIII and XXIII to XXVI collectively, or an oligonucleotide that encodes the HLA peptide. Another embodiment ofthe invention comprises an HLA peptide that occurs at least twice in Tables V-XVIII, or an oligonucleotide that encodes the HLA peptide. Another embodiment ofthe invention comprises an HLA peptide that occurs at least twice in Tables XXIII to XXVI, or an oligonucleotide that encodes the HLA peptide. Another embodiment ofthe invention comprises an HLA peptide that occurs at least once in Tables V-XVIII and is embedded within at least one peptide in Tables XXIH to XXVI, or an oligonucleotide that encodes the HLA peptide.

Another embodiment ofthe invention is antibody epitopes which comprise a peptide region, or an oligonucleotide encoding the peptide region, that has one two, three, four, or five ofthe following characteristics: i) a peptide region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to the full length of that protein in Figure 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Hydrophilicity profile of Figure 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 the 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 the full length of that protein in Figure 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Percent Accessible Residues profile of Figure 7; iv) a peptide region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to the full length of that protein in Figure 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Average Flexibility profile of Figure 8; or v) a peptide region of at least 5 amino acids of a particular peptide of Figure 3, in any whole number increment up to the full length of that protein in Figure 3, that includes an amino acid position having a value equal to or greater than 0.5, 0.6, 0.7, 0.8, 0.9, or having a value equal to 1.0, in the Beta-turn profile of Figure 9. BRIEF DESCRIPTION OF THE FIGURES

Figure 1. The SSH sequences ofthe invention.

Figure 2. Genes and respective encoded proteins ofthe invention.

Figure 3. Amino acid sequences ofthe invention.

Figure 4. Nucleic acid sequence and protein alignments.

Figure 5. Hydrophilicity amino acid profile ofthe proteins set forth in Figure 2 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 the Protscale website (www.expasy.ch/cgi-bin/protscale.pl) through the ExPasy molecular biology server.

Figure 6. Hydropathicity amino acid profile ofthe proteins set forth in Figure 2 determined by computer algorithm sequence analysis using the method of Kyte 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) through the ExPasy molecular biology server.

Figure 7. Percent accessible residues amino acid profile ofthe proteins set forth in Figure 2 determined by computer algorithm sequence analysis using the method of Janin (Janin J., 1979 Nature 277:491-492) accessed on the ProtScale website (www.expasy.ch/cgi-bin/protscale.pl) through the ExPasy molecular biology server.

Figure 8. Average flexibility amino acid profile ofthe proteins set forth in Figure 2 determined by computer algorithm sequence analysis using the method of Bhaskaran and Ponnuswamy (Bhaskaran R., and Ponnuswamy P.K., 1988. Int. J. Pept. Protein Res. 32:242-255) accessed on the ProtScale website (www.expasy.ch/cgi-bin/protscale.pl) through the ExPasy molecular biology server.

Figure 9. Beta-turn amino acid profile ofthe proteins set forth in Figure 2 determined by computer algorithm sequence analysis using the method of Deleage and Roux (Deleage, G., Roux B. 1987 Protein Engineering 1:289-294) accessed on the ProtScale website (www.expasy.ch cgi-bin/protscale.pl) through the ExPasy molecular biology server.

Figure 10. Secondary structure predictions for the proteins set forth in Figure 2.. The secondary structures ofthe proteins set forth in Figure 2 were predicted using the HNN - Hierarchical Neural Network method (Guermeur, 1997, at World Wide Web URL pbil.ibcp.fr/cgi-bin/npsa_automat.pl?page=npsa_ι_n.html), accessed from the ExPasy molecular biology server (at World Wide Web URL www.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 ofthe protein in a given secondary structure is also listed for each variant.

Transmembrane predictions for the proteins set forth in Figure 2. Schematic representations ofthe probability of existence of transmembrane regions and orientation ofthe proteins of Figure 2 based on the TMpred algorithm of Hofmann and Stoffel which utilizes TMB ASE (K. Hofmann, W. Stoffel. TMBASE - A database of membrane spanning protein segments Biol. Chem. Hoppe-Seyler 374:166, 1993). Schematic representation ofthe probability ofthe existence of transmembrane regions and the extracellular and intracellular orientation ofthe proteins of Figure 2 based on the TMHMM algorithm of Sonnhammer, von Heijne, and Krogh (Erik L.L. Sonnhammer, Gunnar von Heijne, and Anders Krogh: A hidden Markov model for predicting transmembrane helices in protein sequences. In Proc. of Sixth Int. Conf. on Intelligent Systems for Molecular Biology, p l75-182 Ed J. Glasgow, T. Littlejohn, F. Major, R. Lathrop, D. Sankoff, and C. Sensen Menlo Park, CA: AAAI Press, 1998). The TMpred and TMHMM algorithms are accessed from the ExPasy molecular biology server (at World Wide Web URL .expasy.ch/tools/). Figure 11. The nucleotide sequences of transcript variants ofthe invention.

Figure 12. This Figure shows amino acid sequences of proteins translated from the corresponding transcript variants set forth in Figure 11.

Figure 13. This Figure displays the alignment of he nucleotide sequences of respective transcript variants.

Figure 14. This Figure displays the alignment ofthe protein sequences from the respective transcript variants. The sub-numbering nomenclature of Figure 11 through Figure 14 is set forth in the following legend:

Figure 15. Expression of 74P3B3 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), two prostate metastasis to lymph node (LN) harvested from two different patients, prostate cancer pool, bladder cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 74P3B3, was performed at 26 and 30 cycles of amplification. Results show strong expression of 74P3B3 in the two prostate metastasis to LN specimens and in prostate cancer pool. Expression was also detected in bladder cancer pool, cancer metastasis pool, and vital pool 2 but not in the vital pool 1.

Figure 16. Expression of 74P3B3 in normal tissues. Two multiple tissue northern blots (A and B; Clontech) both with 2 μg of mRNA/lane, and a LAPC xenograft blot with 10 μg of total RNA/lane (C) were probed with the 74P3B3 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of approximately 7 kB 74P3B3 transcript in prostate but not in the other normal tissues tested. Expression was also detected in LAPC-4AD and LAPC-4AI but not in LAPC-9AD and LAPC-9AI.

Figure 17. Expression of 74P3B3 in prostate cancer patient specimens. RNA was extracted from normal prostate (NP), pool of 3 prostate cancer patient tumors (T) and their normal adjacent tissues (N). Northern blot with 10 μg of total RNA lane was probed with 74P3B3 SSH sequence. Size standards in kilobases (kb) are indicated on the side. The results show strong expression of 74P3B3 in normal prostate and in patient prostate cancer specimens.

Figure 18. Expression of 74P3B3 in patient cancer specimens. Expression of 74P3B3 was assayed in a panel of human cancers (T) and their respective matched normal tissues (N) on RNA dot blots. Upregulated expression of 74P3B3 in tumors compared to normal tissues was observed in prostate, kidney, breast and colon tumors. The expression detected in normal adjacent tissues (isolated from diseased tissues) but not in normal tissues (isolated from healthy donors) may indicate that these tissues are not fully normal and that 74P3B3 may be expressed in early stage tumors.

Figure 19. Expression of 83P4B8 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 83P4B8, was performed at 30 cycles of amplification. Results show strong expression of 83P4B8 in all cancer pools tested. Very low expression was detected in the vital pools.

Figure 20. Expression of 83P4B8 in normal tissues. Two multiple tissue northern blots (A and B; Clontech) both with 2 μg of mRNA lane, and a LAPC xenograft blot with 10 jug of total RNA lane (C) were probed with the 83P4B8 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of two approximately 4.4kb 83P4B8 transcripts in testis and to lower level in thymus but not in the other normal tissues tested. Expression was also detected in all 4 LAPC prostate cancer xenografts.

Figure 21. Expression of 83P4B8 in patient cancer specimens and normal tissues. RNA was extracted from a pool of three prostate cancers (PC), bladder cancers (BC), kidney cancers (KC), colon cancers (CC), lung cancers (LC), ovary cancers (OC), cancer metastasis (Met), pancreas cancers (PaC), as well as from normal prostate (NP), normal bladder (NB), normal kidney (NK), normal colon (NC), normal lung (NL), normal breast (NBr) normal ovary (NO) and normal pancreas (NPa). Northern blot with 10 μg of total RNA lane was probed with 83P4B8 sequence. Size standards in kilobases (kb) are indicated on the side. Results show expression of 83P4B8 in the bladder cancers and ovary cancers. Expression of 83P4B8 was also detected in prostate cancers, kidney cancers, colon cancers, lung cancers, cancer metastasis and pancreas cancer but not in the normal tissues tested.

Figure 22. Expression of 83P4B8 in prostate cancer patient specimens. RNA was extracted from normal prostate (NP), prostate cancer patient tumors (T) and their normal adjacent tissues (N). Northern blot with the side. The results show strong expression of 83P4B8 in the patient prostate cancer specimens.

Figure 23. Expression of 83P4B8 in colon cancer patient specimens. RNA was extracted from colon cancer cell lines (CL), normal colon (N), colon cancer patient tumors (T) and their normal adjacent tissues (Nat). Northern blots with 10 μg of total RNA were probed with the 83P4B8 SSH fragment. Size standards in kilobases are indicated on the side. Results show strong expression of 83P4B8 in the colon tumor tissues and in all three colon cancer cell lines tested, but not in the normal tissues.

Figure 24. Expression of 109P1D4 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool, and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi- quantitative PCR, using primers to 109P1D4, was performed at 30 cycles of amplification. Results show strong expression of 109P1D4 in all cancer pools tested. Very low expression was detected in the vital pools

Figure 25. Expression of 109P1D4 in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA/lane, were probed with the 109P1D4 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of approximately 10 kb 109P1D4 transcript in ovary. Weak expression was also detected in placenta and brain, but not in the other normal tissues tested.

Figure 26. Expression of 109P1D4 in human cancer cell lines. RNA was extracted from a number of human prostate and bone cancer cell lines. Northern blots with 10 μg of total RNA/lane were probed with the 109P1D4 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of 109P1D4 in LAPC-9AD, LAPC-9AI, LNCaP prostate cancer cell lines, and in the bone cancer cell lines, SK-ES-1 and RD-ES.

Figure 27. Expression of 109P1D4 in patient cancer specimens. Expression of 109P1D4 was assayed in a panel of human cancers (T) and their respective matched normal tissues (N) on RNA dot blots. Upregulated expression of 109P1D4 in tumors compared to normal tissues was observed in uterus, lung and stomach. The expression detected in normal adjacent tissues (isolated from diseased tissues) but not in normal tissues (isolated from healthy donors) may indicate that these tissues are not fully normal and that 109P1D4 may be expressed in early stage tumors.

Figure 28. Expression of 151P1C7A by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC prostate cancer xenograft pool (LAPC- 4AD, LAPC-4AI, LAPC-9AD and LAPC-9AI), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 151P1C7A, was performed at 26 and 30 cycles of amplification. Results show strong expression of 151 P 1 C7 A in bladder, lung, and metastasis cancer pools tested. Expression was also detected in xenograft, prostate, kidney and colon cancer pools but not in the vital pools.

Figure 29. Expression of 151P1C7A in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA lane, were probed with the 151P1C7A SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of an approximately 2 kb 151P1C7A transcript in placenta but not in the other normal tissues tested.

Figure 30. Expression of 151P1C7A in bladder cancer patient specimens. RNA was extracted from bladder cancer cell lines (CL; UM-UC-3, J82, SCaBER), normal bladder (Nb), bladder cancer patient tumors (T) and their normal adjacent tissue (N) isolated from bladder cancer patients. Northern blots with lυ μg ol total KISA were probed with the 151P1C7A SSH sequence. Size standards in kilobases are indicated on the side. Results show expression of 15 IP 1 C7A in patient bladder cancer tissues, and in all bladder cancer cell lines tested, but not in normal bladder.

Figure 31. Expression of 151P1C7A in prostate cancer patient specimens. RNA was extracted from normal prostate (NP), prostate cancer patient tumors (T) and their normal adjacent tissues (N). Northern blot with 10 μg of total RNA lane was probed with 151P1C7A SSH sequence. Size standards in kilobases (kb) are indicated on the side. Results show expression of 151P1C7A in the patient prostate cancer specimens.

Figure 32. Expression of 151P4E11 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC prostate cancer xenograft pool (LAPC- 4AD, LAPC-4AI, LAPC-9AD and LAPC-9AI), prostate cancer pool, bladder cancer pool, colon cancer pool, lung cancer pool, ovary 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 151P4E 11 , was performed at 26 and 30 cycles of amplification. Results show strong expression of 151P4E11 in all cancer pools tested. Expression was detected in vital pool 2 but not in vital pool 1.

Figure 33. Expression of 151P4E11 in normal tissues. Two multiple tissue northern blots (A and B; Clontech) both with 2 μg of mRNA/lane, and a LAPC xenograft blot with 10 μg of total RNA/lane (C) were probed with the 151P4E11 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of an approximately 1.2 kb 151P4E11 transcript in prostate, testis, colon and small intestine. Expression was also detected in all the LAPC prostate cancer xenografts LAPC-4AD, LAPC-4AI, and LAPC-9AI, but not in LAPC-9AD.

Figure 34. Expression of 154P2A8 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 154P2A8, was performed at 26 and 30 cycles of amplification. Results show strong expression of 154P2A8 in bladder cancer pool and lung cancer pool. Expression was also detected in prostate cancer pool, kidney cancer pool, colon cancer pool, and cancer metastasis pool but not in vital pool 1 and vital pool 2.

Figure 35. Expression of 156P1D4 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC prostate cancer xenograft pool (LAPC- 4AD, LAPC-4AI, LAPC-9AD and LAPC-9AI), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary 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 156P1D4, was performed at 26 and 30 cycles of amplification. Results show strong expression of 156P1D4 in kidney cancer pool and vital pool 1. Expression was also detected in xenograft pool, prostate cancer pool, bladder cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool and vital pool 2.

Figure 36. Expression of 156P1D4 in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA lane, were probed with the 156P1D4 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of an approximately 2 kb 156P1D4 transcript in kidney and prostate but not in the other normal tissues tested. Figure 37. Expression of 156P1D4 in kidney cancer patient specimens. RNA was extracted from normal kidney (Nk), kidney cancer patient tumors (T) and their normal adjacent tissues (N). Northern blots with 10 μg of total RNA were probed with the 156P1D4 SSH fragment. Size standards in kilobases are indicated on the side. Results show strong expression of 156P1D4 in all kidney tumor tissues tested. The expression of 156P1D4 detected in tumor tissues is stronger than in normal tissues.

Figure 38. Expression of 156P5C12 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC prostate cancer xenograft pool (LAPC- 4AD, LAPC-4AI, LAPC-9AD and LAPC-9AI), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary 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 156P5C12, was performed at 26 and 30 cycles of amplification. Results show strong expression of 156P5C12 in kidney cancer pool and vital pool 1. Expression was also detected in xenograft pool, prostate cancer pool, bladder cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool and vital pool 2.

Figure 39. Expression of 156P5C12 in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA lane, were probed with the 156P5C12 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of an approximately 1.4 kb 156P5C12 transcript in kidney but not in the otiier normal tissues tested.

Figure 40. Expression of 156P5C12 in kidney cancer patient specimens. RNA was extracted from kidney cancer cell lines (CL; 769-P, A498, SW839), normal kidney (N), kidney cancer patient tumors (T) and their normal adjacent tissues (NAT). Northern blots with 10 μg of total RNA were probed with the 156P5C12 SSH fragment. Size standards in kilobases are indicated on the side. Results show expression of 156P5C12 in normal tissues, and in some but not all kidney tumor tissues. Expression was absent in the kidney cancer cell lines tested.

Figure 41. Expression of 159P2B5 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), and bladder cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 159P2B5, was performed at 26 and 30 cycles of amplification. Results show expression of 159P2B5 in bladder cancer pool tested but not in the vital pools.

Figure 42. Expression of 159P2B5 in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA/lane, were probed with the 159P2B5 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show very weak expression of an approximately 4.5 kbl59P2B5 transcript in spleen, kidney and small intestine.

Figure 43. Expression of 159P2B5 in bladder cancer patient specimens. RNA was extracted from bladder cancer cell lines (CL; UM-UC-3, J82, SCaBER), normal bladder (NB), and bladder cancer patient tumors (T) isolated from bladder cancer patients. Northern blots with 10 μg of total RNA were probed with the 159P2B5 SSH sequence. Size standards in kilobases are indicated on the side. Results show expression of 159P2B5 in patient bladder cancer tissues, and in the SCaBER bladder cancer cell line, but not in normal bladder, nor in the other cancer cell lines tested.

Figure 44. Expression of 161P2B7A by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate metastasis to lymph node (LN), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 161P2B7A, was performed at 26 and 30 cycles of amplification. Results show strong expression of 161P2B7A in lung cancer pool and pancreas cancer pool. Expression was also detected in prostate metastasis to LN, prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, ovary cancer pool, breast cancer pool, and cancer metastasis pool. Very low expression was observed in vital pool 2 but not in vital pool 1.

Figure 45. Expression of 161P2B7A in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA/lane, were probed with the 161P2B7A SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show very low expression of 161P2B7A in testis but not in the other normal tissues tested.

Figure 46. Expression of 161P2B7A in Multiple Normal Tissues. An mRNA dot blot containing 76 different samples from human tissues was analyzed using a 161P2B7A SSH probe. Expression was not detected in any ofthe 76 normal tissues tested. The positive genomic DNA control showed very strong signal confirming the validity ofthe experiment.

Figure 47. Expression of 161P2B7A in kidney cancer patient specimens. RNA was extracted from normal kidney (Nk), kidney cancer patient tumors (T) and their normal adjacent tissues (N) isolated from kidney cancer patients. Northern blots with 10 μg of total RNA were probed with the 161P2B7A SSH fragment. Size standards in kilobases are indicated on the side. Results show expression of two 161P2B7A transcripts, approximately 1.2 and 7 kb, in kidney cancer specimens but not in normal kidney.

Figure 48. Expression of 161P2B7A in lung cancer patient specimens. RNA was extracted from lung cancer cell lines (CL), normal lung, lung tumors (T), and their normal adjacent tissues (NAT) isolated from lung cancer patients. Northern blot with 10 μg of total RNA/lane was probed with the 161P2B7A fragment. Size standards in kilobases (kb) are indicated on the side. The results show expression of 161P2B7A in the lung tumors, but not in normal lung tissues. Expression was also detected in the lung cancer cell lines CALU-1, A427 and NCI- 146 but not in the small cell lung cancer cell line NCI-H82.

Figure 49. Expression of 161P2B7A in pancreas and ovary cancer patient specimens. RNA was extracted from normal pancreas (NPa), pancreas cancer (PC), normal ovary (NO), and ovary cancer patient specimen (OC). Northern blot with 10 μg of total RNA lane was probed with the 161P2B7A fragment. Size standards in kilobases (kb) are indicated on the side. The results show expression of 161P2B7A in the pancreas and ovary cancer patient specimens, but not in the normal tissues.

Figure 50. Expression of 179P3G7 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), bladder cancer pool, kidney cancer pool, lung cancer pool, breast cancer pool, cancer metastasis pool, pancreas cancer pool and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 179P3G7, was performed at 26 and 30 cycles of amplification. Results show strong expression of 179P3G7 in kidney cancer pool and breast cancer pool. Expression was also detected in bladder cancer pool, lung cancer pool, cancer metastasis pool, pancreas cancer pool and prostate metastasis to LN, and vital pool 1, but not in vital pool 2.

Figure 51. Expression of 179P3G7 in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA/lane, were probed with the 179P3G7 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of 179P3G7 strongly in skeletal muscle, and weakly in kidney, liver and heart but not in the other normal tissues tested. Figure 52. Expression of 179P3G7 in kidney cancer patient specimens. RNA was extracted from normal kidney (Nk), kidney cancer patient tumors (T) and their normal adjacent tissues (N) isolated from kidney cancer patients. Northern blots with 10 μg of total RNA were probed with the 179P3G7 SSH fragment. Size standards in kilobases are indicated on the side. Results show expression of 179P3G7 in kidney cancer specimens. Expression of 179P3G7 is stronger in kidney tumors compared to normal kidney tissues.

Figure 53. Expression of 184P3C10B by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC prostate cancer xenograft pool (LAPC- 4 AD, LAPC-4AI, LAPC-9AD and LAPC-9AI), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 184P3C10B, was performed at 26 and 30 cycles of amplification. Results show expression of 184P3C10B in xenograft pool, prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, and cancer metastasis pool. Expression was also detected in vital pool 2 but at a much lower level in vital pool 1.

Figure 54. Expression of 184P3C10B in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA lane, were probed with the 184P3C10B SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of approximately 2.4 and 5 kb 184P3C10B transcripts in placenta and to lower level in colon and small intestine, but not in the other normal tissues tested.

Figure 55. Expression of 184P3C10B in bladder cancer patient specimens. RNA was extracted from bladder cancer cell lines (CL; UM-UC-3, J82, SCaBER), normal bladder (Nb), bladder cancer patient tumors (T) and their normal adjacent tissue (N) isolated from bladder cancer patients. Northern blots with 10 μg of total RNA were probed with the 184P3C10B SSH sequence. Size standards in kilobases are indicated on the side. Results show expression of 184P3 C 10B in patient bladder cancer tissues, and in the bladder cancer cell line SCaBER, but not in normal bladder nor in the other bladder cancer cell lines tested.

Figure 56. Expression of 184P3G10 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC prostate cancer xenograft pool (LAPC- 4AD, LAPC-4AI, LAPC-9AD and LAPC-9AI), bladder cancer pool, kidney cancer pool, colon cancer pool, and lung cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 184P3G10, was performed at 26 and 30 cycles of amplification. Results show strong expression of 184P3G10 in bladder cancer pool, kidney cancer pool, and colon cancer pool. Expression was also detected in xenograft pool, lung cancer pool, vital pool 2 but not in vital pool 1.

Figure 57. Expression of 184P3G10 in normal tissues. Two multiple tissue northern blots (Clontech) both with 2 μg of mRNA/lane, were probed with the 184P3G10 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of two approximately 4.4kb 184P3G10 transcripts in colon and small intestine, but not in the other normal tissues tested.

Figure 58. Expression of 184P3G10 in patient cancer specimens and normal tissues. RNA was extracted from a pool of three bladder cancers, colon cancers, lung cancers, breast cancers, ovary cancers, cancer metastasis, as well as from normal prostate (NP), normal bladder (NB), and normal kidney (NK). Northern blot with 10 μg of total RNA lane was probed with 184P3G10 sequence. Size standards in kilobases (kb) are indicated on the side. Results show strong expression of 184P3G10 in the bladder cancers, colon cancers and ovary cancers. Expression of 184P3G10 was also detected in lung cancers, breast cancers, and cancer metastasis but not in the normal tissues tested. Figure 59. Expression of 184P3G10 in bladder cancer patient specimens. RNA was extracted from bladder cancer cell lines (CL; UM-UC-3, J82, SCaBER), normal bladder (N), bladder cancer patient tumors (T) and their normal adjacent tissue (Nat) isolated from bladder cancer patients. Northern blots with 10 μg of total RNA were probed with the 184P3G10 SSH sequence. Size standards in kilobases are indicated on the side. Results show expression of 184P3G10 in patient bladder cancer tissues, but not in normal bladder nor in the bladder cancer cell lines tested.

Figure 60. Expression of 185P2C9 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate metastasis to lymph node (LN), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 185P2C9, was performed at 30 cycles of amplification. Results show strong expression of 185P2C9 in bladder cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool and pancreas cancer pool. Expression was also detected in prostate metastasis to LN, prostate cancer pool, kidney cancer pool, breast cancer pool, cancer metastasis pool, vital pool 2 but not in vital pool 1.

Figure 61. Expression of 185P2C9 in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA/lane, were probed with the 185P2C9 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of and approximately 8.5 kb 185P2C9 transcript in testis and brain, but not in the other normal tissues tested.

Figure 62. Expression of 185P2C9 in bladder cancer patient specimens. RNA was extracted from bladder cancer cell lines (CL; UM-UC-3, J82, SCaBER), normal bladder (Nb), bladder cancer patient tumors (T) and their normal adjacent tissue (N) isolated from bladder cancer patients. Northern blots with 10 μg of total RNA were probed with the 185P2C9 SSH sequence. Size standards in kilobases are indicated on the side. Results show expression of 185P2C9 in bladder cancer patient tissues, and in the bladder cancer cell lines tested. Expression of 185P2C9 is significantly stronger in bladder tumor tissues compared to normal tissues.

Figure 63. Expression of 185P2C9 in kidney cancer patient specimens. RNA was extracted from kidney cancer cell lines (CL; 769-P, A498, Caki-1), normal kidney (N), kidney cancer patient tumors (T) and their normal adjacent tissues (NAT) isolated from kidney cancer patients. Northern blots with 10 μg of total RNA were probed with the 185P2C9 SSH fragment. Size standards in kilobases are indicated on the side. Results show expression of 185P2C9 in kidney cancer specimens and kidney cancer cell lines, but not in normal kidney.

Figure 64. Expression of 186P1H9 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, cancer metastasis pool, and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 186P1H9, was performed at 26 and 30 cycles of amplification. Results show strong expression of 186P1H9 in kidney cancer pool, colon cancer pool, ovary cancer pool, cancer metastasis pool, and pancreas cancer pool. Expression was also detected in bladder cancer pool, lung cancer pool, vital pool 2 but not in vital pool 1.

Figure 65. Expression of 186P1H9 in normal tissues. Two multiple tissue northern blots (Clontech) both with 2 μg of mRNA lane, were probed with the 186P1H9 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of an approximately 2.6 kb 186P1H9 transcript in testis, spleen, pancreas and brain. Lower expression is also detected in heart, skeletal muscle, prostate, colon and small intestine. Figure 66. Expression of 186P1H9 in patient cancer specimens and normal tissues. RNA was extracted from a pool of three kidney cancers (KC), ovary cancers (OC), cancer metastasis (Met), pancreas cancers (PaC), as well as from normal prostate (NP), normal bladder (NB), and normal kidney (NK), normal colon (NC), normal lung (NL), normal breast (NBr), normal ovary (NO), and normal pancreas (NPa). Northern blot with 10 μg of total RNA/lane was probed with 186P1H9 sequence. Size standards in kilobases (kb) are indicated on the side. Results show strong expression of 186P1H9 in the bladder cancers, ovary cancers, cancer metastasis and pancreas cancers, but not in normal tissues. Expression of 186P 1 H9 is significantly stronger in patient cancer tissues compared to normal tissues.

Figure 67. Expression of 186P1H9 in kidney cancer patient specimens. RNA was extracted from kidney cancer cell lines (CL; 769-P, A498, Caki-1), normal kidney (N), kidney cancer patient tumors (T) and their normal adjacent tissues (NAT) isolated from kidney cancer patients. Northern blots with 10 μg of total RNA were probed with the 186P1H9 SSH fragment. Size standards in kilobases are indicated on the side. Results show strong expression of 186P1H9 in kidney cancer patient specimens, but not in normal kidney, nor in the kidney cancer cell lines.

Figure 68. Expression of 186P1H9 in ovarian and testicular cancer patient specimens. RNA was extracted from normal ovary (NO), ovary cancer patient specimens (PI, P2, P3), normal testis (NTe), and testis cancer patient specimens (P4, P5, P6). Northern blot with 10 μg of total RNA lane was probed with the 186P1H9 SSH fragment. Size standards in kilobases (kb) are indicated on the side. The results show strong expression of 186P1H9 in the ovary cancer patient specimens, but not in the normal ovary. Expression was also detected in normal and in testis cancer specimens.

Figure 69. Expression of 187P3F2 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), kidney cancer pool, and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 187P3F2, was performed at 26 and 30 cycles of amplification. Results show strong expression of 187P3F2 in kidney cancer pool, pancreas cancer pool and vital pool 1, but not in vital pool 2.

Figure 70. Expression of 187P3F2 in normal tissues. Two multiple tissue northern blots (Clontech) both with 2 μg of mRNA/lane, were probed with the 187P3F2 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of an 4.5 kb 187P3F2 transcript in kidney and brain, but not in the other tissues tested.

Figure 71. Expression of 187P3F2 in patient cancer specimens and normal tissues. RNA was extracted from a pool of three kidney cancers (KC), pancreas cancers (PaC), as well as from normal prostate (NP), normal bladder (NB), and normal kidney (NK), normal colon (NC), normal lung (NL), normal breast (NBr), normal ovary (NO), and normal pancreas (NPa). Northern blot with 10 μg of total RNA/lane was probed with 187P3F2 sequence. Size standards in kilobases (kb) are indicated on the side. Results show strong expression of 187P3F2 in kidney cancers, pancreas cancers, and normal kidney, but not in the other normal tissues.

Figure 72. Expression of 187P3F2 in pancreas cancer patient specimens. RNA was extracted from pancreas cancer cell lines (CL), normal pancreas (N), and pancreas tumor tissues (T) isolated from pancreatic cancer patients. Northern blot with 10 μg of total RNA/lane was probed with the 187P3F2 SSH fragment. Size standards in kilobases (kb) are indicated on the side. The results show strong expression of 187P3F2 in the pancreas cancer specimens, but not in normal pancreas nor in the cancer cell lines tested. Figure 73. Expression of 192P2G7 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate cancer pool, bladder cancer pool, kidney cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool, pancreas cancer pool, and prostate metastasis to lymph node (LN). Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 186P1H9, was performed at 26 and 30 cycles of amplification. Results show strong expression of 186P1H9 in pancreas cancer pool and prostate metastasis to LN. Expression was also detected in prostate cancer pool, bladder cancer pool, kidney cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool, vital pool 2 but not in vital pool 1.

Figure 74. Expression of 185P3C2 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), and bladder cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 185P3C2, was performed at 26 and 30 cycles of amplification. Results show strong expression of 185P3C2 in bladder cancer pool. Low level expression was detected in vital pool 2, but not in vital pool 1.

DETAILED DESCRIPTION OF THE INVENTION Outline of Sections I.) Definitions

II.) Polynucleotides of the Invention

H.A.) Uses Polynucleotides of the Invention

H.A.I.) Monitoring of Genetic Abnormalities H.A.2.) Antisense Embodiments H.A.3.) Primers and Primer Pairs

II.A.4.) Isolation of Nucleic Acid Molecules that Encode Proteins of the Invention H.A.5.) Recombinant Nucleic Acid Molecules and Host-Vector Systems HI.) Proteins of the Invention

HLA.) Motif-bearing Protein Embodiments HI.B.) Expression of Figure 2-related Proteins HI.C.) Modifications of Figure 2-related Proteins IH.D.) Uses of Figure 2-related Proteins IV.) Antibodies of the Invention V.) Cellular Immune Responses of the Invention VI.) Transgenic Animals of the Invention

VII.) Methods for the Detection of a Gene or Protein of the Invention Viπ.) Methods for Monitoring the Status of Genes and Proteins of the Invention IX.) Identification of Molecules That Interact With the Proteins of Figure 2 X.) Therapeutic Methods and Compositions X.A.) Anti-Cancer Vaccines

X.B.) A Protein of Figure 2 as a Target for Antibody-Based Therapy X.C.) A Protein of Figure 2 as a Target for Cellular Immune Responses X.C.1. 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 H L Peptides X.D.) Adoptive Immunotherapy X.E.) Administration of Vaccines for Therapeutic or Prophylactic Purposes

XI.) Diagnostic and Prognostic Embodiments of the Invention

XII.) Inhibition of the Function of a Protein of the Invention

Xπ.A.) Inhibition of a Protein of Figure 2 with Intracellular Antibodies

Xπ.B.) Inhibition of a Protein of Figure 2 with Recombinant Proteins

XII.C.) Inhibition of Transcription or Translation in Accordance with the Invention

XII.D.) General Considerations for Therapeutic Strategies HI.) 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, terms with commonly understood meanings are defined herein for clarity and/or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art. Many ofthe techniques and procedures described or referenced herein are well understood and commonly employed using conventional methodology by those skilled in the art, such as, for example, the widely utilized molecular cloning methodologies described in Sambrook et al, Molecular Cloning: A Laboratory Manual 2nd. edition (1989) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. As appropriate, procedures involving the use of 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 the American Urological Association (AUA) system, stage CI - C2 disease under the Whitmore-Jewett system, and stage T3 - T4 and N+ disease under the TNM (tumor, node, metastasis) system. In general, surgery is not recommended for patients with locally advanced disease, and these patients have substantially less favorable outcomes compared to patients having clinically localized (organ-confined) prostate cancer. Locally advanced disease is clinically identified by palpable evidence of induration beyond the lateral border ofthe prostate, or asymmetry or induration above the prostate base. Locally advanced prostate cancer is presently diagnosed pathologically following radical prostatectomy if the tumor invades or penetrates the prostatic capsule, extends into the surgical margin, or invades the seminal vesicles.

"Altering the native glycosylation pattern" is intended for purposes herein to mean deleting one or more carbohydrate moieties found in native sequence ofthe genes set forth in Figure 2 (either by removing the underlying glycosylation site or by deleting the glycosylation by chemical and/or enzymatic means), and/or adding one or more glycosylation sites that are not present in the native sequence of a protein set forth in Figure 2. In addition, the phrase includes qualitative changes in the glycosylation ofthe native proteins, involving a change in the nature and proportions ofthe various carbohydrate moieties present.

The term "analog" refers to a molecule wliich is structurally similar or shares similar or corresponding attributes with another molecule (e.g. a protein of Figure 2). For example an analog of a protein of Figure 2 can be specifically bound by an antibody or T cell that specifically binds to the respective protein of Figure 2. The term "antibody" is used in the broadest sense. Therefore an "antibody" can be naturally occurring or man- made such as monoclonal antibodies produced by conventional hybridoma technology. Antibodies ofthe invention comprise monoclonal and polyclonal antibodies as well as fragments containing the antigen-binding domain and/or one or more complementarity determining regions of these antibodies that specifically bind a protein of Figure 2.

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

The term "codon optimized sequences" refers to nucleotide sequences that have been optimized for a particular host species by replacing any codons having a usage frequency of less than about 20%. Nucleotide sequences that have been optimized for expression in a given host species by elimination of spurious polyadenylation sequences, elimination of 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 an "expression enhanced sequences."

The term "cytotoxic agent" refers to a substance that inhibits or prevents the expression activity of cells, function of cells and/or causes destruction of cells. The term is intended to include radioactive isotopes chemotherapeutic agents, and toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof. Examples of cytotoxic agents include, but are not limited to maytansinoids, yttrium, bismuth, ricin, ricin A-chain, doxorubicin, daunorubicin, taxol, ethidium bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxy anthracin 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, calicheamicin, sapaonaria officinalis inhibitor, and glucocorticoid and other chemotherapeutic agents, as well as radioisotopes such as At²¹¹, I¹³¹, 1¹²⁵, Y⁹⁰, Re¹⁸⁶, Re¹⁸⁸, Sm¹⁵³, Bi²¹², P³² and radioactive isotopes of Lu. Antibodies may also be conjugated to an anti-cancer pro-drug activating enzyme capable of converting the 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 al, 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 μg/ml 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 that correspond or are complementary to genes other than the genes of Figure 2 or that encode polypeptides other than proteins of Figure 2 product or fragments thereof. A skilled artisan can readily employ nucleic acid isolation procedures to obtain an isolated polynucleotide. A protein is said to be "isolated," for example, when physical, mechanical or chemical methods are employed to remove a protein of Figure 2 from cellular constituents that are normally associated with the protein. A skilled artisan can readily employ standard purification methods to obtain an isolated Figure 2 protein. Alternatively, an isolated protein can be prepared by chemical means.

The term "mammal" refers to any organism classified as a mammal, including mice, rats, rabbits, dogs, cats, cows, horses and humans. In one embodiment ofthe invention, the mammal is a mouse. In another embodiment ofthe invention, the 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 the case with locally advanced prostate cancer, surgery is generally not indicated for patients with metastatic disease, and hormonal (androgen ablation) therapy is a preferred treatment modality. Patients with 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 the spine, followed by die femur, pelvis, rib cage, skull and humerus. Other common sites for metastasis include lymph nodes, lung, liver and brain. Metastatic prostate cancer is typically diagnosed by open or laparoscopic pelvic lymphadenectomy, whole body radionuclide scans, skeletal radiography, and/or bone lesion biopsy.

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

A "motif, as in biological motif of a Figure 2-related protein, refers to any pattern of amino acids forming part ofthe 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 that is correlated with being immunogenic, either Immorally or cellularly. A motif can be either contiguous or capable of being aligned to certain positions that are generally coπelated with a certain function or property. In the context of HLA motifs, "motif refers to the pattern of residues in a peptide of defined length, usually a peptide of from about 8 to about 13 amino acids for a class I HLA motif and from about 6 to about 25 amino acids for a class II HLA motif, which is recognized by a particular HLA molecule. Peptide motifs for HLA binding are typically different for each protein encoded by each human HLA allele and differ in the pattern ofthe primary and secondary anchor residues.

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

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

The term "polynucleotide" means a polymeric form of nucleotides of at least 10 bases or base pairs in length, either 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 the art, this term if often used interchangeably with "oligonucleotide". A polynucleotide can comprise a nucleotide sequence disclosed herein wherein thymine (T), as shown for example in Figure 2, can also be uracil (U); this definition pertains to the differences between the chemical structures of DNA and RNA, in particular the observation that one ofthe four major bases in RNA is uracil (U) instead of thymine (T). The term "polypeptide" means a polymer of at least about 4, 5, 6, 7, or 8 amino acids. Throughout the specification, standard three letter or single letter designations for amino acids are used. In the art, this 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 the immunogenic peptide and the HLA molecule. One to three, usually two, primary anchor residues within a peptide of defined length generally defines a "motif for an immunogenic peptide. These residues are understood to fit in close contact with peptide binding groove of an HLA molecule, with their side chains buried in specific pockets ofthe binding groove. In one embodiment, for example, the primary anchor residues for an HLA class I molecule are located at position 2 (from the amino terminal position) and at the carboxyl terminal position of a 8, 9, 10, 11, or 12 residue peptide epitope in accordance with the invention. In another embodiment, for example, the primary anchor residues of a peptide that will bind an HLA class II molecule are spaced relative to each other, rather than to the termini of a peptide, where the peptide is generally of at least 9 amino acids in length. The primary anchor positions for each motif and supermotif are set forth in Table IV. For example, analog peptides can be created by altering die presence or absence of particular residues in the primary and/or secondary anchor positions shown in Table IV. Such analogs are used to modulate the binding affinity and/or population coverage of a peptide comprising a particular HLA motif or supermotif.

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

Non-limiting examples of small molecules include compounds that bind or interact with the proteins of Figure 2;, ligands including hormones, neuropeptides, chemokines, odorants, phospholipids, and functional equivalents thereof that bind and preferably inhibit function of a Figure 2 protein. 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, a Figure 2 protein; and are not found in naturally occurring metabolic pathways; and/or are more soluble in aqueous than non-aqueous solutions

"Stringency" of hybridization reactions is readily determinable by one of ordinary skill in the art, and generally is an empirical calculation dependent upon probe length, washing temperature, and salt concentration. In general, longer probes require higher temperatures for proper annealing, while shorter probes need lower temperatures. Hybridization generally depends on the ability of denatured nucleic acid sequences to reanneal when complementary strands are present in an environment below their melting temperature. The higher the degree of desired homology between the probe and hybridizable sequence, the higher the relative temperature that can be used. As a result, it follows that higher relative temperatures would tend to make the reaction conditions more stringent, while lower temperatures less so. For additional details and explanation of stringency of hybridization reactions, see Ausubel et al, Current Protocols in Molecular Biology, Wiley Interscience Publishers, (1995).

"Stringent conditions" or "high stringency conditions", as defined herein, are identified by, but not limited to, those that: (1) employ low ionic strength and high temperature for washing, for example 0.015 M sodium chloride/0.0015 M sodium citrate/0.1% sodium dodecyl sulfate at 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/0.1 % polyvinylpyrrolidone/50 mM sodium phosphate buffer at pH 6.5 with 750 mM 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 x Denhardt's solution, sonicated salmon sperm DNA (50 μg/ml), 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 al, 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 moderately stringent conditions is overnight incubation at 37°C in a solution comprising: 20% formamide, 5 x SSC (150 mM NaCl, 15 mM frisodium citrate), 50 mM sodium phosphate (pH 7.6), 5 x Denhardt's solution, 10% dexfran sulfate, and 20 mg/mL 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 the 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 ofthe 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 ofthe invention. There are numerous embodiments of such vaccines, such as a cocktail of one or more individual peptides; one or more peptides ofthe 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 1-150 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, or 150 or more peptides ofthe invention. The peptides or polypeptides can optionally be modified, such as by lipidation, addition of targeting or other sequences. HLA class I peptides ofthe 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 exhibits a variation from a described type or norm, such as a protein that has one or more different amino acid residues in the corresponding position(s) of a specifically described protein (e.g. a protein of Figure 2 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 "genes of Figure 2-related proteins" ofthe invention include those specifically identified herein, as well as allelic variants, conservative substitution variants, analogs and homologs that can be isolated/generated and characterized without undue experimentation following the methods outlined herein or readily available in the art. Fusion proteins that combine parts of different genes set forth in Figure 2 proteins ofthe invention or fragments thereof, ^■ as well as fusion proteins of a gene of Figure 2 protein and a heterologous polypeptide are also included. Such genes of Figure 2 proteins are collectively referred to as the genes of Figure 2-related proteins, the proteins ofthe invention, or proteins of Figure 2. The term " genes of Figure 2-related protein" refers to a polypeptide fragment or a Figure 2 protein sequence of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or more than 25 amino acids; or, at least 30, 35, 40, 45, 50, 55, 60, 65, 70, 80, 85, 90, 95, 100 or more than 100 amino acids. In certain cases the phrase "corresponding to" or "respective" is used instead ofthe term "-related."

11.1 Polynucleotides of the Invention

One aspect ofthe invention provides polynucleotides corresponding or complementary to all or part of: a gene of Figure 2; gene of Figure 2-related mRNA, a coding sequence of a gene of Figure 2, an open reading frame of a gene of Figure 2, each ofthe foregoing preferably in isolated form. Polynucleotides ofthe invention include polynucleotides encoding Figure 2-related proteins and fragments thereof, DNA, RNA, DNA RNA hybrid, and related molecules, polynucleotides or oligonucleotides complementary to a Figure 2 gene or mRNA sequence or a part thereof, and polynucleotides or oligonucleotides that hybridize to a Figure 2 gene, mRNA, or to a Figure 2 encoding polynucleotide (collectively, "Figure 2 polynucleotides"). In all instances when referred to in this section, T can also be U in Figure 2.

Embodiments of a Figure 2 polynucleotide include: a Figure 2 polynucleotide having the sequence shown in Figure 2, the nucleotide sequence ofthe genes of Figure 2 as shown in Figure 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 ofthe Figure 2 nucleotides comprise, without limitation:

(1) a polynucleotide comprising, consisting essentially of, or consisting of a sequence as shown in Figure 2 (SEQ ID NO: ), wherein T can also be U;

(2) a polynucleotide comprising, consisting essentially of, or consisting of a sequence as shown in

Figure 2 (SEQ ID NOs: ), from the first nucleotide residue of a reading frame through the last nucleotide residue of that reading frame, optionally followed by a stop codon, wherein T can also be U;

(3) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2A.1 (SEQ ID NO: ), from nucleotide residue number 289 through nucleotide residue number

828, optionally followed by a stop codon, wherein T can also be U;

(4) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2A.2 (SEQ ID NO: ), from nucleotide residue number 756 through nucleotide residue number

1439, optionally followed by a stop codon, wherein T can also be U;

(5) . a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2B (SEQ ID NO: ), from nucleotide residue number 25 through nucleotide residue number

4008, optionally followed by a stop codon, wherein T can also be U;

(6) a.polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2C (SEQ ID, NO: ), from nucleotide residue number 846 through nucleotide residue number

3908, optionally followed by a stop codon, wherein T can also be U;

(7) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2D (SEQ ID NO: ), from nucleotide residue number 103 through nucleotide residue number

900, optionally followed by a stop codon, wherein T can also be U; (8) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2E (SEQ ID NO: ), from nucleotide residue number 3 through nucleotide residue number 371, optionally followed by a stop codon, wherein T can also be U;

(9) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2F (SEQ ID NO: ), from nucleotide residue number 250 through nucleotide residue number

1323, optionally followed by a stop codon, wherein T can also be U;

(10) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2G (SEQ ID NO: ), from nucleotide residue number 24 through nucleotide residue number

599, optionally followed by a stop codon, wherein T can also be U;

(11) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2H (SEQ ID NO: ), from nucleotide residue number 178 through nucleotide residue number

858, optionally followed by a stop codon, wherein T can also be U;

(12) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 21 (SEQ ID NO: ), from nucleotide residue number 1517 through nucleotide residue number

2188, optionally followed by a stop codon, wherein T can also be U;

(13) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2J (SEQ ID NO: ), from nucleotide residue number 198 through nucleotide residue number

767, optionally followed by a stop codon, wherein T can also be U;

(14) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2K (SEQ ID NO: ), from nucleotide residue number 72 through nucleotide residue number

1097, optionally followed by a stop codon, wherein T can also be U;

(15) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2L (SEQ ID NO: ), from nucleotide residue number 118 through nucleotide residue number

1233, optionally followed by a stop codon, wherein T can also be U;

(16) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2M (SEQ ID NO: ), from nucleotide residue number 14 through nucleotide residue number

2257, optionally followed by a stop codon, wherein T can also be U;

(17) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2N.1 (SEQ ID NO: ), from nucleotide residue number 140 through nucleotide residue number

4060, optionally followed by a stop codon, wherein T can also be U;

(18) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2N.2 (SEQ ID NO: ), from nucleotide residue number 140 through nucleotide residue number

3565, optionally followed by a stop codon, wherein T can also be U; (19) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2N.3 (SEQ ID NO: ), from nucleotide residue number 140 through nucleotide residue number

4075, optionally followed by a stop codon, wherein T can also be U;

(20) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2(0) (SEQ ID NO: ), from nucleotide residue number 3 through nucleotide residue number

1655, optionally followed by a stop codon, wherein T can also be U;

(21) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2P (SEQ ID NO: ), from nucleotide residue number 170 through nucleotide residue number

1459, optionally followed by a stop codon, wherein T can also be U;

(22) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2Q (SEQ ID NO: ), from nucleotide residue number 60 through nucleotide residue number

1559, optionally followed by a stop codon, wherein T can also be U;

(23) a polynucleotide comprising, consisting essentially of, or consisting ofthe sequence as shown in

Figure 2R (SEQ ID NO: ), from nucleotide residue number 84 through nucleotide residue number

938, optionally followed by a stop codon, wherein T can also be U;

(24) a polynucleotide that encodes a Figure 2-related protein that is at least 90% homologous to an entire amino acid sequence shown in Figure 2A-R (SEQ ID NO: );

(25) a polynucleotide that encodes a Figure 2-related protein that is at least 90% identical to an entire amino acid sequence shown in Figure 2A-R (SEQ ID NO: );

(26) a polynucleotide that encodes at least one peptide set forth in Tables V-XVIII, Table XX, or Tables XXIII to XXVI;

(27) a polynucleotide that encodes a peptide region of at least five amino acids of a peptide of Figure 3 in any whole number increment up to the entire length of that protein, that includes an amino acid position having a value greater than 0.5 in the Hydrophilicity profile of Figure 5 for that protein,;

(28) a polynucleotide that encodes a peptide region of at least five amino acids of a peptide of Figure 3 in any whole number increment up to the entire length ofthe protein, that includes an amino acid position having a value less than 0.5 in the Hydropathicity profile of Figure 6 for that protein;

(29) a polynucleotide that encodes a peptide region of at least five amino acids of a peptide of Figure 3 in any whole number increment up to the entire length ofthe protein, that includes an amino acid position having a value greater than 0.5 in the Percent Accessible Residues profile of Figure 7for that protein;

(30) a polynucleotide that encodes a peptide region of at least 5 amino acids of a peptide of Figure 3 in any whole number increment up to the entire length of that protein, that includes an amino acid position having a value greater than 0.5 in the Average Flexibility profile of Figure 8 for that protein; (31) a polynucleotide that encodes a peptide region of at least 5 amino acids of a peptide of Figure 3 in any whole number increment up to the entire length ofthe protein, that includes an amino acid position having a value greater than 0.5 in the Beta-turn profile of Figure 9 for that protein;

(32) a polynucleotide that encodes a Figure 2-related protein whose sequence is encoded by the cDNAs contained in the plasmid 74P3B3 that was deposited with American Type Culture Collection (ATCC) as Accession No. PTA-1892 on 19 May 2000;

(33) a polynucleotide that is fully complementary to a polynucleotide of any one of (l)-(32);

(34) a polynucleotide that selectively hybridizes under stringent conditions to a polynucleotide of (1) to (33);

(35) a peptide that is encoded by any of (1)— (32); and,

(36) a polynucleotide of any of (l)-(34)or peptide of (35) 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, i.e., integer positions, thereof.

Typical embodiments ofthe invention disclosed herein include the proteins of Figure 2 polynucleotides that encode specific portions ofthe Figure 2 mRNA sequences (and those which are complementary to such sequences) such as those that encode the proteins and/or fragments thereof, for example:

4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 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, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 505, 510, 515, 520, 525, 530, 535, 540, 545, 550, 555, 560, 565, 570, 575, 580, 585, 590, 595, 600, 605, 610, 615, 620, 625, 630, 635, 640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690, 695, 700, 705, .710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760, 765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 815, 820, 825, 830, 835, 840, 845, 850, 855, 860, 865, 870, 875, 880, 885, 890, 895, 900, 905, 910, 915, 920, 925, 930, 935, 940, 945, 950, 955, 960, 965, 970, 975, 980, 985, 990, 995, 1000, 1025, 1050, 1075, 1100, 1125, 1150, 1175, 1200, etc., or more contiguous amino acids of a peptide ofthe invention.

For example, representative embodiments ofthe invention disclosed herein include: polynucleotides and their encoded peptides themselves encoding about amino acid 1 to about amino acid 10 of a Figure 2 protein or variants thereof, polynucleotides encoding about amino acid 10 to about amino acid 20 of a Figure 2 protein or variants thereof, polynucleotides encoding about amino acid 20 to about amino acid 30 of a Figure 2 protein or variants thereof, polynucleotides encoding about amino acid 30 to about amino acid 40 of a Figure 2 protein or variants thereof, polynucleotides encoding about amino acid 40 to about amino acid 50 of a Figure 2 protein or variants thereof, polynucleotides encoding about amino acid 50 to about amino acid 60 of a Figure 2 protein or variants thereof, polynucleotides encoding about amino acid 60 to about amino acid 70 of a Figure 2 protein or variants thereof, polynucleotides encoding about amino acid 70 to about amino acid 80 of a Figure 2 protein or variants thereof, polynucleotides encoding about amino acid 80 to about amino acid 90 of a Figure 2 protein or variants thereof, polynucleotides encoding about amino acid 90 to about amino acid 100 of a Figure 2 protein or variants thereof, or encoding regions from about amino acid 100 to amino acids later in the sequence, in increments of about 10 amino acids, ending at the carboxyl terminal amino acid of a protein ofthe invention, e.g. a protein set forth in Figure 2 or Figure 3. Accordingly polynucleotides encoding portions ofthe amino acid sequence (in increments of about 10 amino acids), of amino acids 100 through the carboxyl terminal amino acid of a Figure 2 protein are embodiments ofthe 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 Figure 2 protein are also within the scope ofthe 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 a Figure 2 protein or variants thereof can be generated by a variety of techniques well known in the art. These polynucleotide fragments can include any portion of gene ofthe invention as shown, e.g., in Figure 2.

Additional illustrative embodiments ofthe invention disclosed herein include a protein of Figure 2 polynucleotide fragments encoding one or more ofthe biological motifs contained within a Figure 2 protein sequence or a variant sequence thereof, including one or more ofthe motif-bearing subsequences of a Figure 2 protein or variant, e.g., set forth in Tables V-XVIII, Table XX, and/or Tables XXIII to XXVI. In another embodiment, typical polynucleotide fragments ofthe invention encode one or more ofthe regions of a Figure 2 protein or variant thereof that exhibit homology to a known molecule. In another embodiment ofthe invention, typical polynucleotide fragments encode one or more ofthe Figure 2 proteins or variants N-glycosylation sites, cAMP and cGMP-dependent protein kinase phosphorylation sites, casein kinase II phosphorylation sites or N- myristoylation site and amidation sites (see, e.g., Table XX).

π.A.. Uses Polynucleotides of the Invention π.A.l._ Monitoring of Genetic Abnormalities

The polynucleotides ofthe preceding paragraphs have a number of different specific uses. The human genes set forth in Figure 2 maps to the chromosomal locations set forth in Example 3. For example, because a Figure 2 gene map to a particular chromosome, polynucleotides that encode different regions ofthe Figure 2 proteins are used to characterize cytogenetic abnormalities of this chromosomal locale, such as abnormalities that are identified as being associated with various cancers. In certain genes, a variety of chromosomal abnormalities including 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) and Finger et al, P.N.A.S. 85(23): 9158-9162 (1988)). Thus, polynucleotides encoding specific regions of the Figure 2 proteins provide new tools that can be used to delineate, with greater precision than previously possible, cytogenetic abnormalities in the chromosomal region that encodes the proteins set forth in Figure 2 that may contribute to the malignant phenotype. In this context, these polynucleotides satisfy a need in the art for expanding the sensitivity of chromosomal screening in order to identify more subtle and less common chromosomal abnormalities (see e.g. Evans et al, Am. J. Obstet. Gynecol 171(4): 1055-1057 (1994)). Furthermore, as the genes set forth in Figure 2 are shown to be-highly expressed in cancers, the Figure 2 polynucleotides are used in methods assessing the status ofthe Figure 2 gene products in normal versus cancerous tissues. Typically, polynucleotides that encode specific regions ofthe Figure 2 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 ofthe Figure 2 genes, such as regions containing one or more motifs. Exemplary assays include both RT-PCR assays as well as single-strand conformation polymorphism (SSCP) analysis (see, 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.

H.A.2.. Antisense Embodiments

Other specifically contemplated nucleic acid related embodiments ofthe 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 a gene set forth in Figure 2. For example, antisense molecules can be RNAs or other molecules, including peptide nucleic acids (PNAs) or non-nucleic acid molecules such as phosphorothioate 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 Figure 2 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., a gene of Figure 2. See for example, Jack Cohen, Oligodeoxynucleotides, Antisense Inhibitors of Gene Expression, CRC Press, 1989; and Synthesis 1:1-5 (1988). The Figure 2 antisense oligonucleotides ofthe present invention include derivatives such as S-oligonucleotides (phosphorothioate 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 ofthe phosphate group is replaced by a sulfur atom. The S-oligos ofthe present invention can be prepared by treatment ofthe corresponding O-oligos with 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. 112:1253-1254 (1990).' Additionally, the Figure 2 antisense oligonucleotides ofthe present invention include morpholino antisense oligonucleotides known in the art (see, e.g., Partridge et al, 1996, Antisense & Nucleic Acid Drug Development 6: 169-175).

The Figure 2 antisense oligonucleotides ofthe present invention typically can be RNA or DNA that is complementary to and stably hybridizes with the first 100 5' codons or last 1003' codons of a genomic sequence or the corresponding mRNA ofthe invention. Absolute complementarity is not required, although high degrees of complementarity are preferred. Use of an oligonucleotide complementary to this region allows for the selective hybridization to mRNA ofthe invention and not to mRNA specifying other regulatory subunits of protein kinase. In one embodiment, the Figure 2 antisense oligonucleotides ofthe present invention are 15 to 30-mer fragments of the antisense DNA molecule that have a sequence that hybridizes to mRNA ofthe invention. Optionally, a Figure 2 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 a gene set forth in Figure 2. Alternatively, the antisense molecules are modified to employ ribozymes in the inhibition of expression of a gene set forth in Figure 2, see, e.g., L. A. Couture & D. T. Stinchcomb; Trends Genet 12: 510-515 (1996). π.A.3.. Primers and Primer Pairs Further specific embodiments ofthe nucleotides ofthe invention include primers and primer pairs, which allow the specific amplification of polynucleotides ofthe invention or of any specific parts thereof, and probes that selectively or specifically hybridize to nucleic acid molecules ofthe 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 Figure 2 polynucleotide in a sample and as a means for detecting a cell expressing a Figure 2 protein.

Examples of such probes include polynucleotides comprising all or part of a human gene set forth in Figure 2. Examples of primer pairs capable of specifically amplifying an mRNA ofthe invention are also disclosed herein. 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 an mRNA of he invention.

The Figure 2 polynucleotides ofthe invention are useful for a variety of purposes, including but not limited to their use as probes and primers for the amplification and or detection ofthe Figure 2 gene(s), mRNA(s), or fragments thereof; as reagents for the diagnosis and/or prognosis of prostate cancer and other cancers; as coding sequences capable of directing the expression of a Figure 2 polypeptide; as tools for modulating or inhibiting the expression of a Figure 2 gene(s) and/or translation of a Figure 2 franscript(s); and as therapeutic agents.

The present invention includes the use of any probe as described herein to identify and isolate a gene set forth in Figure 2 or Figure 2-related nucleic acid sequence ofthe invention 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.

II.A.4.^") Isolation of Nucleic Acid Molecules that Encode Proteins of the Invention

The cDNA sequences described herein, see, e.g., Figure 2, enable the isolation of other polynucleotides encoding gene produces) ofthe invention, as well as the isolation of polynucleotides encoding homologs of protein of Figure 2, alternatively spliced isoforms, allelic variants, and mutant forms of agene product of a gene ofthe invention as well as polynucleotides that encode analogs ofthe Figure 2-related proteins. Various molecular cloning methods that can be employed to isolate full length cDNAs encoding a Figure 2 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 and 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 a Figure 2 gene cDNA can be identified by probing with a labeled cDNA of Figure 2 or a fragment thereof. For example, in one embodiment, a Figure 2 cDNA or a portion thereof is synthesized and used as a probe to retrieve overlapping and full-length cDNAs corresponding to a gene set forth in Figure 2. A gene set forth in Figure 2 itself can be isolated by screening genomic DNA libraries, bacterial artificial chromosome libraries (BACs), yeast artificial chromosome libraries (YACs), and the like, with a respective gene in Figure 2 DNA probe or primer.

H.A.5.. Recombinant Nucleic Acid Molecules and Host-Vector Systems

The invention also provides recombinant DNA or RNA molecules containing a polynucleotide, a fragment, analog or homologue thereof in accordance with the invention, 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 fransformed 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 polynucleotide (fragment, analog or homologue thereof) in accordance with the invention 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 an Sf9 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 a protein in Figure 2 or a fragment, analog or homolog thereof can be used to generate Figure 2 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 Figure 2 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 (Invifrogen) and the retroviral vector pSRαtkneo (Muller et al, 1991, MCB 11:1785). Using these expression vectors, proteins set forth in Figure 2 can be expressed in several prostate cancer and non-prostate cell lines, including for example 293, 293T, rat-1, NIH 3T3 and TsuPrl. The host-vector systems ofthe invention are useful for the production of a Figure 2 protein or fragment thereof. Such host-vector systems can be employed to study the functional properties of proteins set forth in Figure 2 and ofthe proteins of Figure 2 mutations or analogs.

Recombinant human proteins ofthe invention, e.g., set forth in Figure 2, or an analog or homolog or fragment thereof can be produced by mammalian cells transfected with a construct containing a Figure 2-related nucleotide. For example, 293T cells can be transfected with an expression plasmid encoding a protein of Figure 2 or fragment, analog or homolog thereof, a Figure 2-related protein is expressed in the 293T cells, and the recombinant protein ofthe invention is isolated using standard purification methods (e.g., affinity purification using antibodies ofthe invention, e.g., an antibody that specifically binds a protein ofthe invention such as one set forth in Figure 2). In another embodiment, a Figure 2 coding sequence is subcloned into the retroviral vector pSRoJVISVtkneo and used to infect various mammalian cell lines, such as NTH 3T3, TsuPrl, 293 and rat-1 in order to establish cell lines that express a protein ofthe invention. Various other expression systems well known in the art can also be employed. Expression constructs encoding a leader peptide joined in frame to a Figure 2 coding sequence can be used for the generation of a secreted form of recombinant Figure 2 proteins.

As discussed herein, redundancy in the genetic code permits variation in the gene sequences set forth in Figure.2. 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 known sequences ofthe 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 www.dna.affrc.go.jp/~nakamura/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, fransposon-like repeats, and or other such well-characterized sequences that are deleterious to gene expression. The GC content ofthe 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 ofthe open reading frame, as described in Kozak, Mol. Cell Biol, 9:5073-5080 (1989). Skilled artisans understand that the general rule that eukaryotic ribosomes initiate translation exclusively at the 5' proximal AUG codon is abrogated only under rare conditions (see, e.g., Kozak PNAS 92(7): 2662-2666, (1995) and Kozak NAR 15(20): 8125-8148 (1987)).

IH.) Proteins of the Invention

Another aspect ofthe present invention provides Figure 2-related proteins, i.e., proteins ofthe invention. Specific embodiments of Figure 2-related proteins comprise a polypeptide having all or part ofthe amino acid sequence of a human protein set forth in Figure 2. Alternatively, embodiments of Figure 2 proteins comprise variant, homolog or analog polypeptides that have alterations in their amino acid sequence relative to a protein set forth in Figure 2.

In general, naturally occurring allelic variants of a protein set forth in Figure 2 shares a high degree of structural identity and homology (e.g., 90% or more homology). Typically, allelic variants of a Figure 2 protein contain conservative amino acid substitutions within the protein sequences set forth in Figure 2 described herein or contain a substitution of an amino acid from a coπesponding position in a homologue of a protein set forth in Figure 2. One class of Figure 2 allelic variants are proteins that share a high degree of homology with at least a small region of a particular Figure 2 amino acid sequence, but further contain a radical departure from the sequence, such as a non-conservative substitution, truncation, insertion or frame shift. In comparisons of protein sequences, die 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 ofthe protein. Proteins ofthe 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 the environment ofthe particular amino acid and its role in the three-dimensional structure ofthe protein. For example, glycine (G) and alanine (A) can frequently be interchangeable, as can alanine (A) and valine (V). Methionine (M), wliich is relatively hydrophobic, can frequently be interchanged with leucine and isoleucine, and sometimes with valine. Lysine (K) and arginine (R) are frequently interchangeable in locations in which the significant feature ofthe amino acid residue is its charge and the differing pK'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" 2^nd ED. Lubert Stryer ed (Stanford University); Henikoff et al, PNAS 1992 Vol 89 10915-10919; Lei etal, J Biol Chem 1995 May 19; 270(20): 11882-6).

Embodiments ofthe invention disclosed herein include a wide variety of art-accepted variants or analogs of Figure 2 proteins such as polypeptides having amino acid insertions, deletions and substitutions. Figure 2 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 al, Nucl. Acids Res., 13:4331 (1986); Zoller et al, Nucl. Acids Res., 10:6487 (1987)), cassette mutagenesis (Wells et al, Gene, 34:315 (1985)), restriction selection mutagenesis (Wells et al, Philos. Trans. R. Soc. London SerA, 317:415 (1986)) or other known techniques can be performed on the cloned DNA to produce variant DNA in accordance with the invention.

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 prefeπed scanning amino acids are relatively small, neutral amino acids. Such amino acids include alanine, glycine, serine, and cysteine. Alanine is typically a prefeπed scanning amino acid among this group because it eliminates the side-chain beyond the beta-carbon and is less likely to alter the main-chain conformation ofthe variant. Alanine is also typically prefeπed because it is the most common amino acid. Further, it is frequently found in both buried and 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, an isosteric amino acid can be used.

As defined herein, Figure 2 variants, analogs or homologs, have the distinguishing attribute of having at least one epitope that is "cross reactive" with a protein of Figure 2. As used in this sentence, "cross reactive" means that an antibody or T cell that specifically binds to a Figure 2 variant also specifically binds to a Figure 2 protein having an amino acid sequence set forth in Figure 3. A polypeptide ceases to be a variant of a protein shown in Figure 3, when it no longer contains any epitope capable of being recognized by an antibody or T cell that specifically binds to the starting of a Figure 2 protein. Those skilled in the art understand that antibodies that recognize proteins bind to epitopes of varying size, and a grouping ofthe 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 al, J. Immunol 2000 165(12): 6949-6955; Hebbes et al, Mol Immunol (1989) 26(9):865-73; Schwartz et al, J Immunol (1985) 135(4):2598-608.

Other classes of Figure 2-related protein variants share 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more similarity, homology or identity with an amino acid sequence of Figure 3,or a fragment thereof. Another specific class of Figure 2 protein variants or analogs comprise one or more ofthe Figure 2 biological motifs described herein (see, e.g., Table V or XVIII, Table XX, or Tables XXIII to XXVI) or presently known in the art. Thus, encompassed by the present invention are analogs ofthe proteins set forth in Figure 2 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 ofthe art are to be applied to the nucleic or amino acid sequences of Figure 2 or Figure 3.

As discussed herein, embodiments ofthe claimed invention include polypeptides containing less than the full amino acid sequence of a protein shown in Figure 2 or Figure 3. For example, representative embodiments of the invention comprise peptides/proteins having any: 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 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, 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, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465,- 470, 475, 480, 485, 490, 495, 500, 505, 510, 515, 520, 525, 530, 535, 540, 545, 550, 555, 560, 565, 570, 575, 580, 585, 590, 595, 600, 605, 610, 615, 620, 625, 630, 635, 640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690, 695, 700, 705, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760, 765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 815, 820, 825, 830, 835, 840, 845, 850, 855, 860, 865, 870, 875, 880, 885, 890, 895, 900, 905, 910, 915, 920, 925, 930, 935, 940, 945, 950, 955, 960, 965, 970, 975, 980, 985, 990, 995, 1000, 1025, 1050, 1075, 1100, 1125, 1150, 1175, 1200, etc., or more contiguous amino acids of a protein shown in Figure 2 or Figure

3-

Moreover, representative embodiments ofthe invention disclosed herein include polypeptides consisting of about amino acid 1 to about amino acid 10 of a Figure 2 protein shown, polypeptides consisting of about amino acid 10 to about amino acid 20 of a Figure 2 protein, polypeptides consisting of about amino acid 20 to about amino acid 30 of a Figure 2 protein, polypeptides consisting of about amino acid 30 to about amino acid 40 of a Figure 2 protein, polypeptides consisting of about amino acid 40 to about amino acid 50 of a Figure 2 protein, polypeptides consisting of about amino acid 50 to about amino acid 60 of a Figure 2 protein, polypeptides consisting of about amino acid 60 to about amino acid 70 of a Figure 2 protein, polypeptides consisting of about amino acid 70 to about amino acid 80 of a Figure 2 protein, polypeptides consisting of about amino acid 80 to about amino acid 90 of a Figure 2 protein, polypeptides consisting of about amino acid 90 to about amino acid 100 of a Figure 2 protein, etc. throughout the entirety of a protein set forth in Figure 2 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 Figure 2 protein are embodiments ofthe invention. It is to be appreciated that the starting and stopping positions in this paragraph refer to the specified position as well as that position plus or minus 5 residues.

Figure 2-related proteins are generated using standard peptide synthesis technology or using chemical cleavage methods well known in the art. Alternatively, recombinant methods can be used to generate nucleic acid molecules that encode a Figure 2-related protein. In one embodiment, nucleic acid molecules provide a means to generate defined fragments of a Figure 2 protein (or variants, homologs or analogs thereof). πLA.) Motif-bearing Protein Embodiments

Additional illustrative embodiments ofthe invention disclosed herein include polypeptides ofthe invention that comprise the amino acid residues of one or more ofthe biological motifs contained within a protein of Figure 2 polypeptide sequence set forth in Figure 2 or Figure 3. Various motifs are known in the art, and a protein can be evaluated for the presence of such motifs by a number of publicly available Internet sites (see, e.g., World Wide Web URL addresses: pfam.wustl.edu/; searcmauncher.bcm.tmc.edu/seq-search/struc-predict.html; psort.ims.u- tokyo.ac.jp/; www.cbs.dtu.dk/; www.ebi.ac.uk/interpro/scan.html; www.expasy.ch/tools/scnpsitl.html; Epimatrix™ and Epimer™, Brown University, www.brown.edu/Rese__rch/TB-H_V_Lab/φimatrix/epimatι_x.htnn and BIMAS, bimas.dcrt.nih.gov/.). Accordingly, see, e.g., the motif bearing subsequences of all Figure 2 proteins set forth and identified in Tables V to XVIII, Table XX, Table XXI, and Tables XXIII to XXVI. Additionally, Table XIX sets forth several frequently occurring motifs based on pfam searches (see URL address pfam.wustl.edu/). The columns of Table Vffl list (1) motif name abbreviation, (2) percent identity found amongst the different member ofthe 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 ofthe motifs set forth in Tables V to XVIII, Table XX, Table XXI, and Tables XXIII to XXVI are useful in elucidating the specific characteristics of a malignant phenotype in view of the observation that the motifs discussed above are associated with growth dysregulation and because the proteins of Figure 2 are 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 ofthe 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): 1119-1126 (1996); Peterziel 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 al, Exp. Cell Res. 235(1): 145-154 (1997)). Amidation is another protein modification also associated with cancer and cancer progression (see e.g. Treston et al, J. Natl. Cancer Inst. Monogr. (13): 169-175 (1992)).

In another embodiment, proteins ofthe invention comprise one or more ofthe immunoreactive epitopes identified in accordance with art-accepted methods, such as the peptides set forth in Tables V-XVIII and XXIII to XXVI. CTL epitopes can be determined using specific algorithms to identify peptides within a Figure 2 protein that are capable of optimally binding to specified HLA alleles (e.g., Table IV; Epimatrix™ and Epimer™, Brown University, URL www.brown.edu/Rese__rch/TB-H_V_Lab/epirr_atrix/epii___atrix.html and BIMAS, URL bimas.dcrt.nih.gov/.) Moreover, processes for identifying peptides that have sufficient binding affinity for HLA molecules and which are coπelated with being immunogenic epitopes, are well known in the art, and are carried out without undue experimentation. In addition, processes for identifying peptides that are immunogenic epitopes, are well known in the 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 with 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 ofthe 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 prefeπed residue as defined in Table IV; substitute a less-prefeπed residue with a prefeπed residue as defined in Table rV; or substitute an originally-occurring prefeπed residue with another prefeπed residue as defined in Table IV. Substitutions can occur at primary anchor positions or at other positions in a peptide; see, e.g., Table IV.

A variety of references reflect the art regarding the identification and generation of epitopes in a protein of interest as well as analogs thereof. See, for example, WO 9733602 to Chesnut et al; Sette, Immunogenetics 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., Immunogenetics 199745(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)); Kast et al, 1994 152(8): 3904-12; Boπas-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.

Related embodiments ofthe inventions include polypeptides comprising combinations ofthe different motifs set forth in Table X3X; and/or, one or more ofthe predicted CTL epitopes of Tables V to XVIII, and/or, one or more ofthe predicted HTL epitopes of Tables XXIII to XXVI and/or, one or more ofthe T cell binding motifs known in the art. Prefeπed embodiments contain no insertions, deletions or substitutions either within the motifs or the intervening sequences ofthe polypeptides. In addition, embodiments which include a number of either N- terminal and or C-terminal amino acid residues on either side of these motifs may be desirable (to, for example, include a greater portion ofthe polypeptide architecture in which the motif is located). Typically the number of N- terminal and/or C-terminal amino acid residues on either side of a motif is between about 1 to about 100 amino acid residues, preferably 5 to about 50 amino acid residues.

Figure 2-related proteins are embodied in many forms, preferably in isolated form. A purified Figure 2 protein molecule will be substantially free of other proteins or molecules that impair the binding of a protein of Figure 2 to an antibody, T cell or other ligand. The nature and degree of isolation and purification will depend on the intended use. Embodiments of Figure 2-related proteins include purified Figure 2-related proteins and functional, soluble Figure 2-related proteins. In one embodiment, a functional, soluble Figure 2 protein or fragment thereof retains the ability to be bound by an antibody, T cell or other ligand.

The invention also provides Figure 2 proteins comprising biologically active fragments of a Figure 2 amino acid sequence. Such proteins exhibit properties ofthe starting Figure 2 protein, such as the ability to elicit the generation of antibodies that specifically bind an epitope associated with the starting Figure 2 protein; to 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.

Figure 2-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, Kyte-Doolittle, Eisenberg, Karplus-Schultz or Jameson- Wolf analysis, or on the basis of immunogenicity. Fragments that contain such stractures are particularly useful in generating subunit-specific antibodies that bind to a protein of Figure 2, or T cells or in identifying cellular factors that bind to a protein set forth in Figure 2. 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.K., 1988, Int. J. Pept. Protein Res. 32:242-255. Beta-turn profiles can be generated, and immunogenic peptide fragments identified, using the method of Deleage, G., Roux B., 1987, Protein Engineering 1 :289-294.

CTL epitopes can be determined using specific algorithms to identify peptides within a Figure 2 protein that are capable of optimally binding to specified HLA alleles (e.g., by using the SYFPEITHI site at World Wide Web URL sylpeithi.bmi-heidelberg.com/; the listings in Table IV(A)-(E); Epimatrix™ and Epimer™, Brown University, URL (www.brown.edu/Researc_ι/TB-HTV_Lab/epiιra^ and BIMAS, URL bimas.dcrt.nih.gov/).

Illustrating this, peptide epitopes from the proteins set forth in Figure 2 that are presented in the context of human MHC class I molecules HLA-A1, A2, A3, All, A24, B7 and B35 were predicted (Tables V-XVIII). Specifically, the complete amino acid sequence of a Figure 2 protein and relevant portions of other presented variants, i.e., for HLA Class I predictions 9 flanking residues on either side of a point mutation for 10-mers, and for HLA Class II predictions 14 flanking residues on either side of a point mutation for 15-mers, were entered into the HLA Peptide Motif Search algorithm found in the Bioinformatics and Molecular Analysis Section (BIMAS) web site listed above; for HLA Class II the site SYFPEITHI at URL sy_peithi.bmi-heidelberg.com/ was used for HTL epitopes of . Tables XXIII to XXVI.

The HLA peptide motif search algorithm was developed by Dr. Ken Parker based on binding of specific peptide sequences in the groove of HLA Class I molecules, in particular HLA-A2 (see, e.g., 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)). This algorithm allows location and ranking of 8-mer, 9-mer, and 10-mer peptides from a complete protein sequence for predicted binding to HLA-A2 as well as numerous other HLA Class I molecules. Many HLA class I binding peptides are 8-, 9-, 10 or 11-mers. For example, for class I HLA-A2, the epitopes preferably contain a leucine (L) or methionine (M) at position 2 and a valine (V) or leucine (L) at the C- terminus (see, e.g., Parker et al, J. Immunol. 149:3580-7 (1992)). Selected results from a complete protein sequence set forth in Figure 2 that predicted binding peptides are shown in Tables V-XVII. In Tables V-XVIII, the top 50 ranking candidates, 9-mers and 10-mers, for each family member are shown along with their location, the amino acid sequence of each specific peptide, and an estimated binding score. The binding score coπesponds to the estimated half time of dissociation of complexes containing the peptide at 37°C at pH 6.5. Peptides with the highest binding score are predicted to be the most tightly bound to HLA Class I on the cell surface for the greatest period of time and thus represent the best immunogenic targets for T-cell recognition.

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

It is to be appreciated that every epitope predicted by the BIMAS site, Epimer™ and Epimatrix™ sites, or specified by the HLA class I or class II motifs available in the art or which become part ofthe art such as set forth in Table IV (or deteπnined using World Wide Web site URL syfpeithi.bmi-heidelberg.com/, or BIMAS, b-mas.dcrt.nih.gov/) are to be "applied" to a Figure 2 protein in accordance with the invention. As used in this context "applied" means that a Figure 2 protein is evaluated, e.g., visually or by computer-based patterns finding methods, as appreciated by those of skill in the relevant art. Every subsequence of a Figure 2 protein of 8, 9, 10, or 11 amino acid residues that bears an HLA Class I motif, or a subsequence of 9 or more amino acid residues that bear an HLA Class II motif are within the scope ofthe invention.

III.B.) Expression of Figure 2-related Proteins

In an embodiment described in the examples that follow, the proteins set forth in Figure 2 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 a protein of Figure 2 with a C-terminal 6XHis and MYC tag (pcDNA3.1/mycHIS, Invitrogen or Tag5, GenHunter Corporation, Nashville TN). The Tag5 vector provides an IgGK secretion signal that can be used to facilitate the production of a secreted Figure 2 protein in transfected cells. A secreted HIS-tagged Figure 2 protein in the culture media can be purified, e.g., using a nickel column using standard techniques.

IH.C.) Modifications of Figure 2-related Proteins

Modifications of Figure 2-related proteins such as covalent modifications are included within the scope of this invention. One type of covalent modification includes reacting targeted amino acid residues of a protein of Figure 2 polypeptide with an organic derivatizing agent that is capable of reacting with selected side chains or the N- or C- terminal residues of a Figure 2 protein. Another type of covalent modification to a protein of Figure 2 polypeptide included within the scope of this invention comprises altering the native glycosylation pattern of a protein ofthe invention. Another type of covalent modification to a protein of Figure 2 comprises linking a Figure 2 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.

Figure 2-related proteins ofthe present invention can also be modified to form a chimeric molecule comprising a protein of Figure 2 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 ofthe invention fused to another tumor-associated antigen or fragment thereof. Alternatively, a protein in accordance with the invention can comprise a fusion of fragments of a Figure 2 sequence (amino or nucleic acid) such that a molecule is created that is not, through its length, directly homologous to the amino or nucleic acid sequences shown in Figure 2 or Figure 3. Such a chimeric molecule can comprise multiples ofthe same subsequence of a protein set forth in Figure 2. A chimeric molecule can comprise a fusion of a Figure 2-related protein with a polyhistidine epitope tag, which provides an epitope to which immobilized nickel can selectively bind, with cytokines or with growth factors. The epitope tag is generally placed at the amino- or carboxyl- terminus of a Figure 2 protein. In an alternative embodiment, the chimeric molecule can comprise a fusion of a Figure 2-related protein with an immunoglobulin or a particular region of an immunoglobulin. For a bivalent form of the chimeric molecule (also refeπed to as an "immunoadliesin"), such a fusion could be to the Fc region of an IgG molecule. The lg fusions preferably include the substitution of a soluble (transmembrane domain deleted or inactivated) form of a Figure 2 polypeptide in place of at least one variable region within an lg molecule. In a prefeπed embodiment, the immunoglobulin fusion includes the hinge, CH2 and CH3, or the hinge, CHI, CH2 and CH3 regions of an IgGl molecule. For the production of immunoglobulin fusions see, e.g., U.S. Patent No. 5,428,130 issued June 27, 1995.

HI.D.) Uses of Figure 2-related Proteins

The proteins ofthe invention have a number of different specific uses. As the proteins set forth in Figure 2 are highly expressed in one or more cancers, Figure 2-related proteins are used in methods that assess the status of Figure 2 gene products in normal versus cancerous tissues, thereby elucidating the malignant phenotype. Typically, polypeptides from specific regions of a Figure 2 protein are used to assess the presence of perturbations (such as deletions, insertions, point mutations etc.) in those regions (such as regions containing one or more motifs). Exemplary assays utilize antibodies or T cells targeting Figure 2-related proteins comprising the amino acid residues of one or more ofthe biological motifs contained within a protein of Figure 2 polypeptide sequence hi order to evaluate the characteristics of this region in normal versus cancerous tissues or to elicit an immune response to the epitope. Alternatively, Figure 2-related proteins that contain the amino acid residues of one or more ofthe biological motifs in a Figure 2 protein are used to screen for factors that interact with that region ofthe respective protein set forth in Figure 2.

Figure 2 protein fragments/subsequences are particularly useful in generating and characterizing domain- specific antibodies (e.g., antibodies recognizing an extracellular or intracellular epitope of a Figure 2 protein), for identifying agents or cellular factors that bind to a protein in Figure 2 or a particular structural domain thereof, and in various therapeutic and diagnostic contexts, including but not limited to diagnostic assays, cancer vaccines and methods of preparing such vaccines.

Proteins encoded by a gene ofthe invention (e.g., a Figure 2 gene, or analog, homolog or fragment thereof) have a variety of uses, including but not limited to generating antibodies and in methods for identifying ligands and other agents and cellular constituents that bind to a Figure 2 gene product. Antibodies raised against a Figure 2 protein or fragment thereof are useful in diagnostic and prognostic assays, and imaging methodologies in ■ the management of human cancers characterized by expression of a Figure 2 protein, such as those listed in Table I. Such antibodies can be expressed infracellularly and used in methods of treating patients with such cancers. Figure 2-related nucleic acids or proteins are also used in generating HTL or CTL responses.

Various immunological assays useful for the detection of Figure 2 proteins are used, including but not limited to various types of radioimmunoassays, enzyme-linked immunosorbent assays (ELISA), enzyme-linked immunofluorescent assays (ELIFA), immimocytochemical methods, and the like. Antibodies can be labeled and used as immunological imaging reagents capable of detecting cells that express a protein set forth in Figure 2 (e.g., in radioscintigraphic imaging methods). Figure 2 proteins are also particularly useful in generating cancer vaccines, as further described herein.

TV.) Antibodies of the Invention

Another aspect ofthe invention provides antibodies that bind to Figure 2-related proteins. Preferred antibodies specifically bind to a Figure 2-related protein and do not bind (or bind weakly) to peptides or proteins that are not Figure 2-related proteins. For example, antibodies that bind to proteins in Figure 2 can bind to Figure 2-related proteins such as the homologs or analogs thereof.

Antibodies ofthe invention are particularly useful in cancer (see, e.g., the cancers refeπed to in 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 the genes and respective encoded proteins set forth in Figure 2 are also expressed or overexpressed in these other cancers. Moreover, intracellularly expressed antibodies (e.g., single chain antibodies) are therapeutically useful in treating cancers in which the expression of a gene and encoded protein of Figure 2 is involved, such as advanced or metastatic prostate cancers.

The invention also provides various immunological assays useftd for the detection and quantification a protein of Figure 2 and mutants thereof. Such assays can comprise one or more Figure 2 antibodies capable of recognizing and binding a Figure 2-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 radioi munoassays, enzyme-linked immunosorbent assays (ELISA), enzyme-linked immunofluorescent assays (ELIFA), and the like.

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

In addition, immunological imaging methods capable of detecting a cancer expressing a gene ofthe invention are also provided by the invention, including but not limited to radioscintigraphic imaging methods using labeled Figure 2 antibodies. Such assays are clinically useful in the detection, monitoring, and prognosis of a gene ofthe invention- expressing cancer.

Antibodies ofthe invention are also used in methods for purifying a Figure 2-related protein and for isolating proteins ofthe invention, e.g., Figure 2 homologues and related molecules. For example, a method of purifying a Figure 2-related protein comprises incubating a Figure 2 antibody, which has been coupled to a solid matrix, with a lysate or other solution containing a Figure 2-related protein under conditions that permit the antibody to bind to the Figure 2- related protein; washing the solid matrix to eliminate impurities; and eluting the Figure 2-related protein from the coupled antibody. Other uses of antibodies in accordance with the invention include generating anti-idiotypic antibodies that mimic a Figure 2 protein.

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

In addition, naked DNA immunization techniques known in the art are used (with or without a purified Figure 2-related protein or agene of Figure 2-expressing cells) to generate an immune response to the encoded immunogen (for review, see Donnelly et al, 1997, Ann. Rev. Immunol. 15: 617-648). The amino acid sequence of a Figure 2 protein can be analyzed to select specific regions ofthe protein for generating antibodies. For example, hydrophobicity and hydrophilicity analyses of Figure 2 amino acid sequences are used to identify hydrophilic regions in the protein. Regions of a Figure 2 protein that show immunogenic structure, as well as other regions and domains, can readily be identified using various other methods known in the art, such as Chou- Fasman, Garnier-Robson,"Kyte-Doolittle, Eisenberg, Karplus-Schultz or Jameson-Wolf analysis. Hydrophilicity profiles can be generated 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 using the method of Kyte, J. and Doolittle, R.F., 1982, J. Mol. Biol. 157:105-132. Percent (%) Accessible Residues profiles can be generated using the method of Janin J., 1979, Nature 277:491-492. Average Flexibility profiles can be generated using the method of Bhaskaran R., Ponnuswamy P.K., 1988, Int. J. Pept. Protein Res. 32:242-255. Beta-turn profiles can be generated using the method of Deleage, G., Roux B., 1987, Protein Engineering 1:289-294. Thus, each region identified by any of these programs or methods is within the scope of ti e present invention. Metiiods for die generation of antibodies in accordance with the invention are further illustrated by way ofthe Examples provided herein. Methods for preparing a protein or polypeptide for use as an immunogen are well known in the art. Also well known in the art are methods for preparing immunogenic conjugates of a protein with a carrier, such as BSA, KLH or other carrier protein. In some circumstances, direct conjugation using, for example, carbodiimide reagents are used; in other instances linking reagents such as those supplied by Pierce Chemical Co., Rockford, IL, are effective. Administration of protein immunogen is often conducted by injection over a suitable time period and with use of a suitable adjuvant, as is understood in die art. During the immunization schedule, titers of antibodies can be taken to determine adequacy of antibody formation.

Monoclonal antibodies ofthe invention 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 the standard hybridoma technology of Kohler and Milstein or modifications that immortalize antibody-producing B cells, as is generally known. Immortalized cell lines that secrete the desired antibodies are screened by immunoassay in which the antigen is a Figure 2-related protein. When the 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 ofthe invention can also be produced, by recombinant means. Regions that bind specifically to the desired regions of a Figure 2 protein can also be produced in the context of chimeric or coirφlementarity-determining region (CDR) grafted antibodies of multiple species origin. Humanized or human antibodies that specifically bind to a proteins of Figure 2 can also be produced, and are prefeπed for use in therapeutic contexts. Methods for humanizing murine and other non-human antibodies, by substituting one or more ofthe non- human antibody CDRs for coπesponding 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. Nad. Acad. Sci. USA 89: 4285 and 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 al, 1998, Nature Biotechnology 16: 535-539). Fully human monoclonal antibodies ofthe invention can be generated using cloning technologies employing large human lg 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 monoclonal antibodies ofthe invention 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 efficientiy produces high affinity authentic human antibodies.

Reactivity of antibodies ofthe invention with a Figure 2-related protein can be established by a number of well known means, including Western blot, immunoprecipitation, ELISA, and FACS analyses using, as appropriate, Figure 2-related proteins, or protein of Figure 2-expressing cells or extracts thereof. An Figure 2 antibody ofthe invention, 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 an enzyme. Further, bi-specific antibodies specific for two or more epitopes are generated using methods generally known in the art. 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.) Cellular Immune Responses of the Invention

The mechanism by which T cells recognize antigens has been delineated. Efficacious peptide epitope vaccine compositions ofthe invention induce a therapeutic or prophylactic immune responses in very broad segments ofthe world-wide population. For an understanding ofthe 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 (Buus, S. et al, Cell 47:1071, 1986; Babbitt, B. P. et al, Nature 317:359, 1985; Townsend, A. and Bodmer, E..,Annu. Rev. Immunol. 7:601, 1989; Germain, R. N., Annu. Rev. Immunol. 11:403, 1993). Through the study of single amino acid substituted antigen analogs and the sequencing of endogenously bound, naturally processed peptides, critical residues that coπespond to motifs required for specific binding to HLA antigen molecules have been identified and are set forth in Table IV (see also, e.g., Southwood, et al, J. Immunol. 160:3363, 1998; Rammensee, et al, Immunogenetics 41:178, 1995; Rammensee et al, SYFPEITHI, access via World Wide Web at URL syfpeit_ii.bmi-heidelberg.com/; Sette, A. and Sidney, J. Curr. Opin. Immunol. 10:478, 1998; Engelhard, V. H., Curr. Opin. 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 74:929-937, 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 crystallographic analyses of HLA-peptide complexes have revealed pockets within the peptide binding cleft/groove of HLA molecules which accommodate, in an allele-specific mode, residues borne by peptide ligands; these residues in turn determine the HLA binding capacity ofthe peptides in which they are present. (See, e.g., Maddejn, D.R. Annu. Rev. Immunol. 13:587, 1995; Smith, et al, Immunity 4:203, 1996; Fremont et al, Immunity 8:305, 1998; Stern et al, Structure 2:245, 1994; Jones, E.Y. Curr. Opin. Immunol. 9:75, 1997; Brown, J. H. et al, Nature 364:33, 1993; Guo, 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. et al, Science 257:919, 19.92; Saper, M. A. , Bjorkman, P. J. and Wiley, D. C, J. Mol. 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 coπelated 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 ofthe epitope and its coπesponding HLA molecule. Additional confirmatory work can be performed to select, amongst these vaccine candidates, epitopes with prefeπed characteristics in terms of population coverage, and/or immunogenicity.

Various sfrategies can be utilized to evaluate cellular immunogenicity, including:

1) Evaluation of primary T cell cultures from normal individuals (see, e.g., Wentworth, P. A. et al, Mol. Immunol. 32:603, 1995; Celis, E. et al, Proc. Natl. Acad. Sci. USA 91:2105, 1994; Tsai, V. et al, J. Immunol. 158:1796, 1997; Kawashirna, 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 the 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 fransgenic mice (see, e.g., Wentworth, P. A. et al, 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 a<_ωinistered subcutaneously to HLA fransgenic mice. Several weeks following immunization, splenocytes are removed and cultured in vitro in the presence of test peptide for approximately one week. Peptide-specific T cells are detected using, e.g., a ->lCr-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 either 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 7:97, 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 the presence of test peptide plus antigen presenting cells (APC) to allow activation of "memory" T cells, as compared to "naive" T cells. At the end ofthe culture period, T cell activity is detected using assays including ^* lcr release involving peptide-sensitized targets, T cell proliferation, or lymphokine release.

VI.) Transgenic Animals of the Invention

Nucleic acids that encode a Figure 2-related protein can also be used to generate either transgenic animals or "knock out" animals that, in turn, are useful in the development and screening of therapeutically useful reagents. In accordance with established techniques, cDNA encoding a protein of Figure 2 can be used to clone genomic DNA that encodes a protein of Figure 2. The cloned genomic sequences can then be used to generate transgenic animals containing cells that express DNA that encode a Figure 2 protein. Methods for generating transgenic animals, particularly animals such as mice or rats, have become conventional in the art and are described, for example, in U.S. Patent Nos. 4,736,866 issued 12 April 1988, and 4,870,009 issued 26 September 1989. Typically, particular cells would be targeted for a nucleic acid sequence of Figure 2 transgene incorporation with tissue- specific enhancers.

Transgenic animals that include a copy of a transgene encoding a Figure 2 protein can be used to examine the effect of increased expression of DNA that encodes the Figure 2 protein. 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 ofthe 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 homologues of Figure 2 proteins can be used to construct a Figure 2 protein "knock out" animal that has a defective or altered gene encoding the Figure 2 protein as a result of homologous recombination between the endogenous gene encoding the Figure 2 protein and altered genomic DNA encoding the Figure 2 protein, infroduced into an embryonic cell ofthe animal. For example, cDNA that encodes a Figure 2 protein can be used to clone genomic DNA encoding the Figure 2 protein, in accordance with established techniques. A portion ofthe genomic DNA encoding a Figure 2 protein 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 wliich the infroduced DNA has homologously recombined 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 Stem Cells: A Practical Approach, E. J. Robertson, ed. (IRL, Oxford, 1987), pp. 113-152). A chimeric embryo can then be implanted into a suitable pseudopregnant female foster animal, and the embryo brought to term to create a "knock out" animal. Progeny harboring the homologously recombined DNA in their germ cells can be identified by standard techniques and used to breed animals in which all cells ofthe animal contain the homologously recombined 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 protein of Figure 2.

VH.) Methods for the Detection of a Gene or Protein of the Invention

Another aspect ofthe present invention relates to methods for detecting Figure 2 polynucleotides and Figure 2- related proteins, as well as methods for identifying a cell that expresses a gene set forth in Figure 2. The expression profile of a gene or protein set forth in Figure 2 makes it a diagnostic marker for metastasized disease. Accordingly, the status of Figure 2 gene products provides information useful for predicting a variety of factors mcluding susceptibility to advanced stage disease, rate of progression, and/or tumor aggressiveness. As discussed in detail herein, the status of Figure 2 gene products in patient samples can be analyzed by a variety protocols that are well known in the art including immunohistochemical analysis, the variety of Northern blotting techniques including in situ hybridization, RT-PCR analysis (for example on laser capture micro-dissected samples), Western blot analysis and tissue aπay analysis.

More particularly, the invention provides assays for the detection of Figure 2 polynucleotides in a biological sample, such as serum, bone, prostate, and other tissues, urine, semen, cell preparations, and the like. Detectable Figure 2 polynucleotides include, for example, a Figure 2 gene or fragment thereof, a Figure 2 mRNA, alternative splice variants of Figure 2 mRNAs, and recombinant DNA or RNA molecules that contain a Figure 2 polynucleotide. A number of methods for amplifying and/or detecting the presence of Figure 2 polynucleotides are well known in the art and can be employed in the practice of this aspect ofthe invention.

In one embodiment, a method for detecting an a Figure 2 mRNA in a biological sample comprises producing cDNA from the sample by reverse transcription using at least one primer; amplifying the cDNA so produced using Figure 2 polynucleotides as sense and antisense primers to amplify Figure 2 cDNAs therein; and detecting the presence o the amplified Figure 2 cDNA. Optionally, the sequence ofthe amplified Figure 2 cDNA can be determined.

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

The invention also provides assays for detecting the presence of a Figure 2 protein in a tissue or other biological sample such as serum, semen, bone, prostate, urine, cell preparations, and the like. Methods for detecting a Figure 2-related protein are also well known and include, for example, immunoprecipitation, immunohistochemical analysis, Western blot analysis, molecular binding assays, ELISA, ELIFA and the like. For example, a method of detecting the presence of a Figure 2-related protein in a biological sample comprises first contacting the sample with a Figure 2 antibody, a Figure 2-reactive fragment thereof, or a recombinant protein containing an antigen binding region of a Figure 2 antibody; and then detecting the binding of a Figure 2-related protein in the sample.

Methods for identifying a cell that expresses a gene of Figure 2 are also within the scope ofthe invention. In one embodiment, an assay for identifying a cell that expresses a Figure 2 gene comprises detecting the presence of a Figure 2 mRNA in the cell. Methods for the detection of particular mRNAs in cells are well known and include, for example, hybridization assays using complementary DNA probes (such as in situ hybridization using labeled riboprobes to a gene of Figure 2, Northern blot and related techniques) and various nucleic acid amplification assays (such as RT- PCR using complementary primers specific for genes of Figure 2, and other amplification type detection methods, such as, for example, branched DNA, SISBA, TMA and the like). Alternatively, an assay for identifying a cell that expresses a Figure 2 gene comprises detecting the presence of a Figure 2-related protein in the cell or secreted by the cell. Various methods for the detection of proteins are well known in the art and are employed for the detection of Figure 2-related proteins and cells that express Figure 2-related proteins.

Expression analysis of Figure 2 proteins is also useful as a tool for identifying and evaluating agents that modulate Figure 2 gene expressions. For example, Figure 2 gene expression is significantly upregulated in prostate cancer, and is expressed in cancers ofthe tissues listed in Table I. Identification of a molecule or biological agent - that inhibits Figure 2 gene expression or over-expression in cancer cells is of therapeutic value. For example, such an agent can be identified by using a screen that quantifies a Figure 2 gene expression by RT-PCR, nucleic acid hybridization or antibody binding.

VUI.) Methods for Monitoring the Status of Genes and Proteins of the Invention

Oncogenesis is known to be a multistep process where cellular growth becomes progressively dysregulated and cells progress from a normal physiological state to precancerous and then cancerous states (see, e.g., Alers et al, Lab Invest. 77(5): 437-438 (1997) and Isaacs et al, Cancer Surv. 23: 19-32 (1995)). In this context,' examining a biological sample for evidence of dysregulated cell growth (such as abeπant gene of Figure 2 expression in cancers) allows for early detection of such abeπant physiology, before a pathologic state such as cancer has progressed to a stage that therapeutic options are more limited and or the prognosis is worse. In such examinations, the status ofthe genes and proteins in Figure 2 in a biological sample of interest can be compared, for example, to the status of that gene and/or protein of Figure 2 in a coπesponding 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 a gene and/or protein of Figure 2 in the biological sample (as compared to the normal sample) provides evidence of dysregulated cellular growth. In addition to using a biological sample that is not affected by a pathology as a normal sample, one can also use a predetermined normative value such as a predetermined normal level of mRNA expression (see, e.g., Grever et al, J. Comp. Neural. 1996 Dec 9; 376(2): 306-14 and U.S. Patent No. 5,837,501) to compare the status of a gene or protein in a sample.

The term "status" in this context is used according to its art accepted meaning and refers to the condition or state of a gene and its products. Typically, skilled artisans use a number of parameters to evaluate the condition or state of a gene and its products. These include, but are not limited to the location of expressed gene products (including the location of gene of Figure 2 expressing cells) as well as the level, and biological activity of expressed gene products (such as Figure 2 mRNA, polynucleotides and polypeptides). Typically, an alteration in the status of a gene and/or protein of Figure 2 comprises a change in the location of a protein Figure 2 and/or cells that express a protein of Figure 2and/or an increase in Figure 2 mRNA and/or protein expression.

The status in a sample of a gene or protein of Figure 2 can be analyzed by a number of means well known in the art, including without limitation, immunohistochemical analysis, in situ hybridization, RT-PCR analysis on laser capture micro-dissected samples, Western blot analysis, and tissue aπay analysis. Typical protocols for evaluating the status of a Figure 2 gene and gene product are found, for example in Ausubel et al. eds., 1995, Cuπent Protocols In Molecular Biology, Units 2 (Northern Blotting), 4 (Southern Blotting), 15 (Immunoblotting) and 18 (PCR Analysis). Thus, the status of a gene or protein in Figure 2 in a biological sample is evaluated by various methods utilized by skilled artisans including, but not limited to genomic Southern analysis (to examine, for example perturbations in a Figure 2 gene), Northern analysis and/or PCR analysis of Figure 2 mRNA (to examine, for example alterations in the polynucleotide sequences or expression levels of Figure 2 mRNAs), and, Western and/or immunohistochemical analysis (to examine, for example alterations in polypeptide sequences, alterations in polypeptide localization within a sample, alterations in expression levels of Figure 2 proteins and/or associations of Figure 2 proteins with polypeptide binding partners). Detectable Figure 2 polynucleotides include, for example, a Figure 2 gene or fragment thereof, a Figure 2 mRNA, alternative splice variants, Figure 2 mRNAs, and recombinant DNA or RNA molecules containing a Figure 2 polynucleotide.

The expression profile of each gene of Figure 2 makes it a diagnostic marker for local and/or metastasized disease, and provides information on the growth or oncogenic potential of a biological sample. In particular, the status of agene or protein of Figure 2 provides information useful for predicting susceptibility to particular disease stages, progression, and/or tumor aggressiveness. The invention provides methods and assays for deteπiiining the expression or mutational status of a gene of Figure 2 and diagnosing cancers that express a gene of Figure 2, such as cancers ofthe tissues listed in Table I. For example, because each gene of Figure 2 mRNA is highly expressed in cancers relative to normal tissue, assays that evaluate the levels of Figure 2 mRNA transcripts or proteins in a biological sample are used to diagnose a disease associated with dysregulation of a gene set forth in Figure 2, and can provide prognostic information useful in defining appropriate therapeutic options.

The expression status ofthe genes and proteins set forth in Figure 2 provides information including the presence, stage and location of dysplastic, precancerous and cancerous cells, predicting susceptibility to various stages of disease, and/or for gauging tumor aggressiveness. Moreover, the expression profile makes it useful as an imaging reagent for metastasized disease. Consequently, an aspect ofthe invention is directed to the various molecular prognostic and diagnostic methods for exaniining the status of these genes and proteins 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 ofthe genes and proteins in Figure 2 in a biological sample can be examined by a number of well-known procedures in the art. For example, the status ofthe genes and proteins in Figure 2 in a biological sample taken from a specific location in the body can be examined by evaluating the sample for the presence or absence of a Figure 2 protein expressing cells (e.g. those that express Figure 2 mRNAs or proteins). This examination can provide evidence of dysregulated cellular growth, for example, when Figure 2 protein-expressing cells are found in a biological sample that does not normally contain such cells (such as a lymph node), because such alterations in the status ofthe genes and proteins in Figure 2 in a biological sample are often associated with dysregulated cellular growth. Specifically, one indicator of dysregulated cellular growth is the metastases of cancer cells from an organ of origin (such as the prostate) to a different area ofthe body (such as a lymph node). In this context, evidence of dysregulated cellular growth is important for example because occult lymph node metastases can be detected in a substantial proportion of patients with prostate cancer, and such metastases are associated with known predictors of disease progression (see, 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 Figure 2 gene products by determining the status of Figure 2 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 Figure 2 gene products in a coπesponding normal sample. The presence of abeπant Figure 2 gene products in the test sample relative to the normal sample provides an indication ofthe presence of dysregulated cell growth within the cells ofthe individual.

In another aspect, the invention provides assays useful in deteπnining the presence of cancer in an individual, comprising detecting a significant increase in Figure 2 mRNA or protein expression in a test cell or tissue sample relative to expression levels in the coπesponding normal cell or tissue. The presence of Figure 2 mRNA can, for example, be evaluated in tissue samples including but not limited to those listed in Table I. The presence of significant Figure 2 protein expression or over-expression in any of these tissues is useful to indicate the emergence, presence and/or severity of a cancer, where the coπesponding normal tissues do not express'Figure 2 mRNA or express it at lower levels.

In a related embodiment, the genes and proteins in Figure 2 status is determined at the protein level rather than at the nucleic acid level. For example, such a method comprises determining the level of a Figure 2 protein expressed by cells in a test tissue sample and comparing the level so determined to the level of a Figure 2 protein expressed in a coπesponding normal sample. In one embodiment, the presence of a Figure 2 protein is evaluated, for example, using immunohistochemical methods. Antibodies ofthe invention or binding partners capable of detecting a Figure 2 protein expression are used in a variety of assay formats well known in the art for this purpose.

In a further embodiment, one can evaluate the status of Figure 2 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 the nucleotide and amino acid sequences are observed in a large number of proteins associated with a growth dysregulated phenotype (see, e.g., Maπogi et al, 1999, J. Cutan. Pathol. 26(8):369-378). For example, a mutation in the sequence of a Figure 2 gene can indicate the presence or promotion of a tumor. Such assays therefore have diagnostic and predictive value where a mutation in a Figure 2 gene 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, the size and structure of nucleic acid or amino acid sequences of Figure 2, or the gene products of one of these genes are observed by the Northern, Southern, Western, PCR and DNA sequencing protocols as discussed herein. In addition, other methods for observing perturbations in nucleotide and amino acid sequences such as single strand conformation polymorphism analysis are well known in the 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 the methylation status of a Figure 2 gene in a biological sample. Abeπant demethylation and/or hypermethylation of CpG islands in gene 5' regulatory regions frequently occurs in immortalized and transformed cells, and can result in altered expression of various genes. For example, promoter hypermethylation ofthe pi-class glutathione S-transferase (a protein expressed in normal prostate but not expressed in >90% of prostate carcinomas) appears to permanently silence transcription of this gene and is the most frequently detected genomic alteration in prostate carcinomas (De Marzo et al, Am. J. Pathol. 155(6): 1985-1992 (1999)). In addition, this alteration is present in at least 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 ofthe LAGE-I tumor specific gene (which is not expressed in normal prostate but is expressed in 25-50% of prostate cancers) is induced by deoxy-azacytidine in lymphoblastoid cells, suggesting that tumoral expression is due to demethylation (Lethe et al, Int. J. Cancer 76(6): 903-908 (1998)). A variety of assays for examining methylation status of a gene are well known in the art. For example, one can utilize, in Southern hybridization approaches, methylation-sensitive restriction enzymes that cannot cleave sequences that contain methylated CpG sites to assess the methylation status of CpG islands. In addition, MSP (methylation specific PCR) can rapidly profile the methylation status of all the CpG sites present in a CpG island of a given gene. This procedure involves initial modification of DNA by sodium bisulfite (which will convert all unmethylated cytosines to uracil) followed by amplification using primers specific for methylated versus unmethylated DNA. Protocols involving methylation interference can also be found for example in Cuπent Protocols . In Molecular Biology, Unit 12, Frederick M. Ausubel et al. eds., 1995.

Gene amplification is an additional method for assessing the status of a Figure 2 gene. Gene amplification is measured in a sample directly, for example, by conventional Southern blotting or Northern blotting to quantitate the transcription of mRNA (Thomas, 1980, Proc. Natl. Acad. Sci. USA, 77:5201-5205), dot blotting (DNA analysis), or in situ hybridization, using an appropriately labeled probe, based on the sequences provided herein. Alternatively, antibodies are employed that recognize specific duplexes, including DNA duplexes, RNA duplexes, and DNA-RNA hybrid duplexes or DNA-protein duplexes; The antibodies in turn are labeled and the assay carried out where the duplex is bound to a surface, so that upon the formation of duplex on the surface, the presence of antibody bound to the duplex can be detected.

Biopsied tissue or peripheral blood can be conveniently assayed for the presence of cancer cells using for example, Northern, dotblot or RT-PCR analysis to detect expression of a gene of Figure 2. The presence of RT-PCR amplifiable Figure 2 mRNA provides an indication ofthe presence of cancer. RT-PCR assays are well known in the art. RT-PCR detection assays for tumor cells in peripheral blood are cuπently being evaluated for use in the diagnosis and management of a number of human solid tumors. In the prostate cancer field, these include RT-PCR assays for the detection of cells expressing PSA and PSM (Verkaik et al, 1997, Urol. Res. 25:373-384; Ghossein et al, 1995, J. Clin. Oncol. 13:1195-2000; Heston etal, 1995, Clin. Chem 41:1687-1688).

A further aspect ofthe invention is an assessment ofthe susceptibility that an individual has for developing cancer. In one embodiment, a method for predicting susceptibility to cancer comprises detecting Figure 2 mRNA or a protein ofthe invention in a tissue sample, its presence indicating susceptibility to cancer, wherein the degree of Figure 2 mRNA expression coπelates to the degree of susceptibility. In a specific embodiment, the presence of a protein ofthe invention in, e.g., prostate tissue is examined, with the presence of a protein of Figure 2 in the sample providing an indication of prostate cancer susceptibility (or the emergence or existence of a prostate tumor). Similarly, one can evaluate the integrity a gene in Figure 2 nucleotide and amino acid sequences in a biological sample, in order to identify perturbations in the structure of these molecules such as insertions, deletions, substitutions and the like. The presence of one or more perturbations in genes or gene products ofthe invention in the sample is an indication of cancer susceptibility (or the emergence or existence of a tumor).

The invention also comprises methods for gauging tumor aggressiveness. In one embodiment, a method for gauging aggressiveness of a tumor comprises deterrriining the level of Figure 2 mRNA or a Figure 2 protein expressed by tumor cells, comparing the level so determined to the level of Figure 2 mRNA or a Figure 2 protein expressed in a coπesponding normal tissue taken from the same individual or a normal tissue reference sample, wherein the degree of Figure 2 mRNA or a Figure 2 protein expression in the tumor sample relative to the normal sample indicates the degree ■ of aggressiveness. In a specific embodiment, aggressiveness of a tumor is evaluated by determining the extent to which a gene of Figure 2 is expressed in the tumor cells, with higher expression levels indicating more aggressive tumors. Another embodiment is the evaluation ofthe integrity of Figure 2 nucleotide and/or amino acid sequences in a biological sample, in order to identify perturbations in the structure of these molecules such as insertions, deletions, substitutions and the like. The presence of one or more perturbations indicates more aggressive tumors.

Another embodiment of the invention is directed to methods for observing the progression of a malignancy in an individual over time. In one embodiment, methods for observing the progression of a malignancy in an individual over time comprise deterrriining the level of Figure 2 mRNA or a Figure 2 protein expressed by cells in a sample ofthe tumor, comparing the level so determined to the level of Figure 2 mRNA or a Figure 2 protein expressed in an equivalent tissue sample taken from the same individual at a different time, wherein the degree of Figure 2 mRNA or a Figure 2 protein expression in the tumor sample over time provides information on the progression ofthe cancer. In a specific embodiment, the progression of a cancer is evaluated by deteimining Figure 2 gene or protein expression in the tumor cells over time, where increased expression over time indicates a progression ofthe cancer. Also, one can evaluate the integrity of Figure 2 nucleotide and amino acid sequences in a biological sample in order to identify perturbations in the structure of these molecules such as insertions, deletions, substitutions and the like, where the presence of one or more perturbations indicates a progression ofthe cancer.

The above diagnostic approaches can be combined with any one of a wide variety of prognostic and diagnostic . protocols known in the art. For example, another embodiment ofthe invention is directed to methods for observing a coincidence between the expression of a Figure 2 gene and/or Figure 2 gene products (or perturbations in a Figure 2 gene and/or Figure 2 gene products) and a factor that is associated with malignancy, as a means for diagnosing and prognosticating the status of a tissue sample. A wide variety of factors associated with malignancy can be utilized, such ■ as the expression of genes associated with 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 et 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 the expression of a Figure 2 gene and/or Figure 2 gene products (or. perturbations in a Figure 2 gene and/or Figure 2 gene products) and another factor that is associated with malignancy are useful, for example, because the presence of a set of specific factors that coincide with disease provides information crucial for diagnosing and prognosticating the status of a tissue sample.

In one embodiment, methods for observing a coincidence between the expression of a Figure 2 gene and Figure 2 gene products (or perturbations in a Figure 2 gene and/or Figure 2 gene products) and another factor associated with malignancy entails detecting the overexpression of Figure 2 mRNA and/or protein in a tissue sample; detecting the overexpression of PSA mRNA or protein in a tissue sample (or PSCA or PSM expression, etc.), and observing a coincidence of Figure 2 mRNA and/or protein and PSA mRNA or protein overexpression (or PSCA or PSM expression). In a specific embodiment, the expression of a gene of Figure 2 and PSA mRNA in prostate tissue is examined, where the coincidence of a Figure 2 gene and PSA mRNA overexpression in the sample indicates the existence of prostate cancer, prostate cancer susceptibility or the emergence or status of a prostate tumor.

Methods for detecting and quantifying the expression of Figure 2 mRNA or protein are described herein, and standard nucleic acid and protein detection and quantification technologies are well known in the art. Standard methods for the detection and quantification of Figure 2 mRNA include in situ hybridization using labeled Figure 2 gene riboprobes, Northern blot and related techniques using Figure 2 polynucleotide probes, RT-PCR analysis using primers specific for Figure 2 genes, and other amplification type detection methods, such as, for example, branched DNA, SISBA, TMA and the like. In a specific embodiment, semi-quantitative RT-PCR is used to detect and quantify Figure 2 mRNA expression. Any number of primers capable of amplifying a Figure 2 gene can be used for this purpose, including but not limited to the various primer sets specifically described herein. In a specific embodiment, polyclonal or monoclonal antibodies specifically reactive with a wild-type Figure 2 protein can be used in an immunohistochemical assay of biopsied tissue.

IX.) Identification of Molecules That Interact With Proteins of Figure 2

The Figure 2 protein and nucleic acid sequences disclosed herein allow, a skilled artisan to identify proteins, small molecules and other agents that interact with the genes or proteins in Figure 2, as well as pathways activated by genes or proteins in Figure 2 via any one of a variety of art accepted protocols. For example, one can utilize one ofthe so-called interaction trap systems (also refeπed to as the "two-hybrid assay"). In such systems, molecules interact and reconstitute a transcription factor which directs expression of a reporter gene, whereupon the expression ofthe 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 1998 and 6,004,746 issued 21 December 1999. Algorithms are also available in the art for genome-based predictions of protein function (see, e.g., Marcotte, et al, ■ Nature 402: 4 November 1999, 83-86).

Alternatively one can screen peptide libraries to identify molecules that interact with a protein sequence of the invention, e.g., a protein of Figure 2. In such methods, peptides that bind to Figure 2 proteins are identified by screening libraries that 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 a Figure 2 protein(s).

Accordingly, peptides having a wide variety of uses, such as therapeutic, prognostic or diagnostic reagents, are thus identified without any prior information on the structure ofthe expected ligand or receptor molecule. Typical peptide libraries and screening methods that can be used to identify molecules that interact with Figure 2 protein sequences are disclosed for example in U.S. Patent Nos. 5,723,286 issued 3 March 1998 and 5,733,731 issued 31 March 1998.

Alternatively, cell lines that express a protein of Figure 2 are used to identify protein-protein interactions mediated by the respective proteins of Figure 2. Such interactions can be examined using immunoprecipitation techniques (see, e.g., Hamilton B.J., et al. Biochem. Biophys. Res. Commun. 1999, 261:646-51). Figure 2 proteins can be immunoprecipitated from the respective proteins of Figure 2-expressing cell line using antibodies ofthe invention that specifically bind that protein. Alternatively, antibodies against His-tag can be used in a cell line engineered to express fusions of a protein of Figure 2 and a His-tag (vectors mentioned above). The immunoprecipitated complex can be examined for protein association by procedures such as Western blotting, ³⁵S- methionine labeling of proteins, protein microsequencing, silver staining and two-dimensional gel electrophoresis.

Small molecules and ligands that interact with the genes and proteins in Figure 2 can be identified through related embodiments of such screening assays. For example, small molecules can be identified that interfere with protein function, including molecules that interfere with protein ofthe invention'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 tumorigenesis. Similarly, small molecules that modulate a proteins of Figure 2-related ion channel, protein pump, or cell communication functions are identified and used to treat patients that have a cancer that expresses a Figure 2 gene (see, e.g., Hille, B., Ionic Channels of Excitable Membranes 2^nd Ed., Sinauer Assoc, Sunderland, MA, 1992). Moreover, ligands that regulate the function of a protein ofthe invention can be identified based on tiieir ability to bind proteins ofthe invention and activate a reporter construct. Typical methods are discussed for example in U.S. Patent No. 5,928,868 issued 27 July 1999, and include methods for forming hybrid ligands in which at least one ligand is a small molecule. In an illustrative embodiment, cells engineered to express a fusion protein of a Figure 2 protein and a DNA-binding protein are used to co-express a fusion protein of a hybrid ligand/small molecule and a cDNA library transcriptional activator protein. The cells further contain a reporter gene, the expression of which is conditioned on the proximity ofthe first and second fusion proteins to each other, an event that occurs only if the hybrid ligand binds to target sites on both hybrid proteins. Those cells that express the reporter gene are selected and the unknown small molecule or the unknown ligand is identified. This method provides a means of identifying modulators which activate or inhibit a protein of the invention.

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

X.) Therapeutic Methods and Compositions

The identification of a Figure 2 as a protein that is normally expressed in a restricted set of tissues, but which is also expressed in certain cancers, opens a number of therapeutic approaches to the treatment of such cancers. As contemplated herein, the genes and proteins in Figure 2 function as a transcription factor involved in activating tumor-promoting genes or repressing genes that block tumorigenesis.

Accordingly, therapeutic approaches that inhibit the activity of a Figure 2 protein are useful for patients suffering from a cancer that expresses a gene of Figure 2. These therapeutic approaches generally fall into two classes. One class comprises various methods for inhibiting the binding or association of a Figure 2 protein with its binding partner or with other proteins. Another class comprises a variety of methods for inhibiting the franscription of a Figure 2 gene or translation of Figure 2 mRNA.

X.A.) Anti-Cancer Vaccines The invention provides cancer vaccines comprising a Figure 2-related protein or a Figure 2-related nucleic acid. In view ofthe expression of a Figure 2 protein, cancer vaccines prevent and/or treat genes of Figure 2-expressing cancers with minimal or no effects on non-target tissues. The use of a tumor antigen in a vaccine tiiat 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 Figure 2-related protein, or a nucleic acid sequence that encodes a Figure 2-related protein and recombinant vectors capable of expressing and presenting immunogen ofthe invention (which typically comprises a number of antibody or T cell epitopes). Skilled artisans understand that a wide variety of vaccine systems for delivery of immunoreactive epitopes are known in the art (see, e.g., Heryln et al, Ann Med 1999 Feb 31(l):66-78; Maruyama et al, Cancer Immunol Immunother 2000 Jun 49(3):123- 32) Briefly, such methods of generating an immune response (e.g. humoral and/or cell-mediated) in a mammal, comprise the steps of: exposing the mammal's immune system to an immunoreactive epitope (e.g. an epitope present in a protein ofthe invention, e.g., shown in Figure 3 or analog or homolog thereof) so that the mammal generates an immune response that is specific for that epitope (e.g. generates antibodies that specifically recognize that epitope). In a prefeπed method, an immunogen contains a biological motif, see e.g., Tables V-XVIII, Tables XXIII to XXVI; or a peptide of a size range from a protein in Figure 2 indicated in Figure 5, Figure 6, Figure 7, Figure 8, and/or Figure 9.

The entire Figure 2 protein, immunogenic regions or epitopes thereof can be combined and delivered by various means. Such vaccine compositions can include, for example, lipopeptides (e.g.,Vitiello, A. et al, J. Clin. Invest. 95:341, 1995), peptide compositions encapsulated in poly(DL-lactide-co-glycolide) ("PLG") microspheres (see, e.g., Eldridge, et al, Molec. Immunol. 28:287-294, 1991: Alonso et al, Vaccine 12:299-306, 1994; Jones et al, Vaccine 13:675-681, 1995), peptide compositions contained in immune stimulating complexes (ISCOMS) (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., Tam, J. P., Proc. Natl. Acad. Sci. U.S.A. 85:5409-5413, 1988; Tam, 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 (Waπen, H. S., Vogel, F. R., and Chedid, L. A. Annu. Rev. Immunol. 4:369, 1986; Gupta, R. K. et al, Vaccine 11:293, 1993), liposomes (Reddy, R. et al, J. Immunol. 148:1585, 1992; Rock, K. L., Immunol. Today 17:131, 1996), or, naked or particle absorbed cDNA (Ulmer, J. B. et al, Science 259:1745, 1993; Robinson, H. L., Hunt, L. A., and Webster, R. G., Vaccine 11:957, 1993; Shiver, J. W. et al, In: Concepts in vaccine development, Kaufmann, S. H. E., ed., p. 423, 1996; Cease, K. B., and Berzofsky, J. A., Annu. Rev. Immunol. 12:923, 1994 and Eldridge, J. H. et al, Sem. Hematol. 30:16, 1993). Toxin-targeted delivery technologies, also known as receptor mediated targeting, such as those of Avant Immunotherapeutics, Inc. (Needham, Massachusetts) may also be used.

In patients with a protein of Figure 2-associated cancer, the vaccine compositions ofthe 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 a Figure 2 protein that bind coπesponding HLA alleles (see e.g., Table IV; Epimer™ and Epimatrix™, Brown University (URL www.brown.edu Rese_u:ch/IH-HIV_Lab/epiir_a1xix/epimatr .h and, BIMAS, (URL bimas.dcrt.nih.gov/; SYFPEITHI at URL syfpeithi.bmi-heidelberg.com/). In a prefeπed embodiment, an ofthe invention contains one or more amino acid sequences identified using techniques well known in the art, such as the sequences shown in Tables V-XVIII 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 motifsupermotif (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 II molecule, as a convention 15-mer peptides that bind to HLA class II alleles are generally presented (see, e.g., Tables XXIII to XXVI). Due to the binding groove differences between HLA Class I and II, HLA Class I 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 other, not the overall peptide, i.e., additional amino acids can be attached to the amino and/or carboxyl termini of a motif-bearing sequence. HLA Class II epitopes are often 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids long, or longer than 25 amino acids.

Antibody-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 the mammal's immune system to an immunogenic epitope on a protein (e.g. a Figure 2 protein) so that an immune response is generated. A typical embodiment consists of a method for generating an immune response to a protein in Figure 2 in a host, by contacting the host with a sufficient amount of at least one protein in Figure 2 B cell or cytotoxic T-cell epitope or analog thereof; and at least one periodic interval thereafter re-contacting the host with the B cell or cytotoxic T-cell epitope or analog thereof. A specific embodiment consists of a method of generating an immune response against a Figure 2-related protein or a man-made multiepitopic peptide comprising: administering an immunogen of the invention (e.g. a Figure 2 protein or a peptide fragment thereof, an Figure 2 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, J. Immunol. 2000 164(3); 164(3): 1625-1633; Alexander et al, 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 an immunogen ofthe invention by: adniinistering in vivo to muscle or skin ofthe individual's body a DNA molecule that comprises a DNA sequence that encodes an immunogen ofthe invention, the DNA sequence operatively linked to regulatory sequences which control the expression ofthe 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 antiidiotypic antibody can be administered that mimics a protein set forth in Figure 2, in order to generate a response to the target antigen.

Nucleic Acid Vaccines:

Vaccine compositions ofthe invention include nucleic acid-mediated modalities. DNA or RNA that encode protein(s) ofthe 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 Figure 2 proteins. Constructs comprising DNA encoding a Figure 2-related protein/immunogen and appropriate regulatory sequences can be injected directly into muscle or skin of an individual, such that the cells ofthe muscle or skin take-up the construct and express the encoded Figure 2 protein immunogen. Alternatively, a vaccine comprises a Figure 2-related protein. Expression ofthe Figure 2-related protein immunogen results in the generation of prophylactic or therapeutic humoral and cellular immunity against cells that bear the Figure 2-related protein. Various prophylactic and therapeutic genetic immunization techniques known in the art can be used (for review, see information and references published at Internet address www.genweb.com). Nucleic acid-based delivery is described, for instance, in Wolff et. al, Science 247:1465 (1990) as well as U.S. Patent Nos. 5,580,859; 5,589,466; 5,804,566; 5,739,118; 5,736,524; 5,679,647; WO 98/04720. Examples of DNA-based delivery technologies include "naked DNA", facilitated (bupivicaine, polymers, peptide-mediated) delivery, cationic lipid complexes, and particle-mediated ("gene gun") or pressure-mediated delivery (see, e.g., U.S. Patent No. 5,922,687).

For therapeutic or prophylactic immunization purposes, proteins ofthe invention can be expressed via viral or bacterial vectors. Various viral gene delivery systems that can be used in the practice ofthe invention include, but are not limited to, vaccinia, fowlpox, canarypox, adenovirus, influenza, poliovirus, adeno-associated virus, lentivirus, and sindbis virus (see, e.g., Restifo, 1996, Cuπ. Opin. Immunol. 8:658-663; Tsang etal. J. Natl. Cancer Inst. 87:982-990 (1995)). Non-viral delivery systems can also be employed by introducing naked DNA encoding a Figure 2-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. Another vector is BCG (Bacille Calmette Guerin). BCG vectors are described in Stover et al, Nature 351:456-460 (1991). A wide variety of other vectors useful for therapeutic administration or immunization ofthe peptides ofthe invention, e.g. adeno and 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 Figure 2-related nucleic acid molecule. In one embodiment, the ftdl-length human gene of Figure 2 cDNA is employed. In another embodiment, Figure 2 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 QDC) to present antigen of the invention to a patient's immune system. Dendritic cells express MHC class I and II molecules, B7 co-stimulator, and IL-12, and are thus highly specialized antigen presenting cells. In prostate cancer, autologous dendritic cells pulsed with peptides ofthe 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 peptide immunogens ofthe invention to T cells in the context of MHC class I or II molecules. In one embodiment, autologous dendritic cells are pulsed with immunogenic peptides capable of binding to MHC class I and/or class II molecules. In another embodiment, dendritic cells are pulsed with the complete Figure 2 protein. Yet another embodiment involves engineering the overexpression of a Figure 2 gene in dendritic cells using various implementing vectors known in the art, such as adenovirus (Arthur et al, 1997, Cancer Gene Ther. 4:17-25), retrovirus (Henderson et al, 1996, Cancer Res. 56:3763-3770), lentivirus, adeno-associated virus, DNA transfection (Ribas et al, 1997, Cancer Res. 57:2865-2869), or tumor-derived RNA transfection (Ashley et al, 1997, J. Exp. Med. 186:1177-1182). Cells that express proteins ofthe invention can also be engineered to express immune modulators, such as GM-CSF, and used as immunizing agents.

X.B.) A Protein of Figure 2 as a Target for Antibody-based Therapy

Proteins ofthe invention, e.g. Figure 2, are attractive targets for antibody-based therapeutic strategies. A number of antibody strategies are 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 infrabodies). Because ofthe expression profiles ofthe proteins set forth in figure 2, , e.g., expressed by cancer cells of various lineages at higher levels compared to ^• coπesponding normal cells, systemic administration of proteins in Figure 2-immunoreactive compositions are prepared that exhibit excellent sensitivity without toxic, non-specific and/or non-target effects caused by binding of the immunoreactive composition to non-target organs and tissues. Antibodies specifically reactive with domains of Figure 2 proteins are useful to systemically treat cancers that express a protein of Figure 2, either as conjugates with a toxin or therapeutic agent, or as naked antibodies capable of inhibiting cell proliferation or function.

Antibodies ofthe invention can be introduced into a patient such that the antibody binds to a protein ofthe invention and modulate a function, such as an interaction with a binding partner, and consequently mediates destruction ofthe tumor cells and/or inhibits the growth ofthe tumor cells. Mechanisms by which such antibodies exert a therapeutic effect can include complement-mediated cytolysis, antibody-dependent cellular cytotoxicity, modulation ofthe physiological function of proteins ofthe invention, inhibition 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 protein ofthe invention such as a protein 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 that cell (e.g. a protein of Figure 2), 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 the 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 antibody that specifically binds a protein of Figure 2) that binds to a marker (e.g. a protein of Figure 2) 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 a Figure 2 protein, comprising conjugating the cytotoxic agent to an antibody that iminunospecifically binds to a protein in Figure 2 epitope, and, exposing the cell to the antibody-agent conjugate. Another illustrative embodiment is a method of treating an individual suspected of suffering from metastasized cancer, comprising a step of administering parenterally to said individual a pharmaceutical composition comprising a therapeutically effective amount of an antibody conjugated to a cytotoxic and/or therapeutic agent.

Cancer immunotherapy using antibodies o the invention 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 90:3179-3186, 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 (Zhong et al, 1996, Leuk. Res. 20:581-589), colorectal cancer (Moun et al, 1994, Cancer Res. 54:6160- 6166; Velders et al, 1995, Cancer Res. 55:4398-4403), and breast cancer (Shepard et al, 1991, J. Clin. Immunol. 11 : 117-127). Some therapeutic approaches involve conjugation of naked antibody to a toxin or radioisotope, such as the conjugation of Y⁹¹ or I¹³¹ to anti-CD20 antibodies (e.g., Zevalin™, IDEC Pharmaceuticals Corp. or Bexxar™, Coulter Pharmaceuticals), while others involve co-administration of antibodies and other therapeutic agents, such as Herceptin™ (trastuzumab) with paclitaxel (Genentech, Inc.). The antibodies can be conjugated to a therapeutic agent. To treat prostate cancer, for example, antibodies ofthe invention 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 maytansinoid (e.g., taxane-based Tumor-Activated Prodrug, TAP, platform, ImmunoGen, Cambridge, MA, also see e.g., US Patent 5,416,064).

Although antibody therapy directed to a protein of the invention is useful for all stages of cancer, antibody therapy can be particularly appropriate in advanced or metastatic cancers. Treatment with the antibody therapy ofthe invention is indicated for patients who have received one or more rounds of chemotherapy. Alternatively, antibody therapy ofthe invention is combined with a chemotherapeutic or radiation regimen for patients who have not received chemotherapeutic treatment. Additionally, antibody therapy can enable the use of reduced dosages of concomitant chemotherapy, particularly for patients who do not tolerate the toxicity ofthe chemotherapeutic 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 the use of various antibodies together with chemotherapeutic agents. Treatment with the antibody therapy ofthe invention is indicated for patients who have received one or more rounds of chemotherapy. Alternatively, antibody therapy ofthe invention is combined with a chemotherapeutic or radiation regimen for patients who have not received chemotherapeutic treatment. Additionally, antibody therapy can enable the use of reduced dosages of concomitant chemotherapy, particularly for patients who do not tolerate the toxicity ofthe chemotherapeutic agent very well.

Cancer patients can be evaluated for the presence and level of expression of a gene of Figure 2, preferably using immunohistochemical assessments of tumor tissue, quantitative imaging of a protein ofthe invention, or other techniques that reliably indicate the presence and degree of a Figure 2 protein expression. Immunohistochemical analysis of tumor biopsies or surgical specimens is prefeπed for this purpose. Methods for immunohistochemical analysis of tumor tissues are well known in the art.

Monoclonal antibodies ofthe invention that freat cancers (e.g., of a tissue of Table I) include those that initiate a potent immune response against the tumor or those that are directly cytotoxic. In this regard, monoclonal antibodies (mAbs) ofthe invention can elicit tumor cell lysis by either complement-mediated or antibody-dependent cell cytotoxicity (ADCC) mechanisms, both of which require an intact Fc portion ofthe immunoglobulin molecule for interaction with effector cell Fc receptor sites on complement proteins. In addition, mAbs ofthe invention that exert a direct biological effect on tumor growth are useful to treat cancers that express proteins in Figure 2. Mechanisms by which directly cytotoxic mAbs act include: inhibition of cell growth, modulation of cellular differentiation, modulation of tumor angiogenesis factor profiles, and the induction of apoptosis. The mechanism(s) by which a particular mAbs ofthe invention exert 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 monoclonal antibodies, or human/mouse chimeric mAbs can induce moderate to sfrong immune responses against the non-human antibody. This can result in clearance ofthe 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, prefeπed monoclonal antibodies used in the therapeutic methods ofthe invention are those that are either fully human or humanized and that bind specifically to the target of proteins in Figure 2 antigens with high affinity but exhibit low or no antigenicity in the patient.

Therapeutic methods ofthe invention contemplate the administration of single mAbs as well as combinations, or cocktails, of different mAbs. Such mAb cocktails can have certain advantages inasmuch as they contain mAbs that target different epitopes, exploit different effector mechanisms or combine directly cytotoxic mAbs with mAbs that rely on immune effector functionality. Such mAbs in combination can exhibit synergistic therapeutic effects. In addition, mAbs ofthe invention can be administered concomitantly with other therapeutic modalities, including but not limited to various chemotherapeutic agents, androgen-blockers, immune modulators (e.g., IL-2, GM-CSF), surgery or radiation. The mAbs ofthe invention are administered in their "naked" or unconjugated form, or can have a therapeutic agent(s) conjugated to them.

Antibody formulations ofthe invention are administered via any route capable of delivering the antibodies to a tumor cell. Routes of administration include, but are not limited to, intravenous, infraperitoneal, intramuscular, intratumor, intradermal, and the like. Treatment generally involves repeated achninistration of an antibody preparation of the invention, via an acceptable route of adminisfration 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 mg 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 ofthe mAb preparation represents an acceptable dosing regimen. Preferably, the initial loading dose is administered as a 90 minute or longer infusion. The periodic maintenance dose is administered as a 30 minute or longer infusion, provided the initial dose was well tolerated. As appreciated by those of skill in the art, various factors can influence the ideal dose regimen in a particular case. Such factors include, for example, the binding affinity and half life ofthe Ab or mAbs used, the degree of expression ofthe protein ofthe invention in the patient, the extent of circulating shed protein ofthe invention, the desired steady-state antibody concenfration level, frequency of treatment, and the influence of chemotherapeutic or other agents used in combination with the treatment method ofthe invention, as well as the health status of a particular patient.

Optionally, patients should be evaluated for the levels of a protein ofthe invention in a given sample (e.g. the levels of circulating Figure 2 protein antigen and/or proteins of Figure 2-expressing cells) in order to assist in the determination ofthe most effective dosing regimen, etc. Such evaluations are also used for monitoring purposes throughout therapy, and are useful to gauge therapeutic success in combination with the evaluation of other parameters (for example, urine cytology and or ImmunoCyt levels in bladder cancer therapy, or by analogy, serum PSA levels in prostate cancer therapy).

Anti-idiotypic antibodies ofthe invention can also be used in anti-cancer therapy as a vaccine for inducing an immune response to cells that express a Figure 2-related protein. In particular, the generation of anti-idiotypic antibodies is well known in the art; this methodology can readily be adapted to generate anti-idiotypic anti-protein of Figure 2 antibodies that mimic an epitope on a Figure 2-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 al, 1996, Cancer Immunol. Immunother. 43:65-76). Such an anti-idiotypic antibody can be used in cancer vaccine strategies.

X.C.) A Protein of Figure 2 as a Target for Cellular Immune Responses

Vaccines and methods of preparing vaccines that contain an immunogenically effective amount of one or more HLA-binding peptides as described herein are further embodiments ofthe invention. Furthermore, vaccines in accordance with the invention encompass compositions of one or more ofthe claimed peptides. A peptide can be present in a vaccine individually. Alternatively, the peptide can exist as a homopolymer comprising multiple copies ofthe same peptide, or as a heteropolymer of various peptides. Polymers have the advantage of increased immunological reaction and, where different peptide epitopes are used to make up the polymer, the additional ability to induce antibodies and or CTLs that react with different antigenic determinants ofthe 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.

Caπiers that can be used with vaccines ofthe invention are well known in the art, and include, e.g., thyroglobulin, albumins such as human serum albumin, tetanus toxoid, polyamino acids such as poly L-lysine, poly L-glutamic acid, influenza, hepatitis B virus core protein, and the 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 the art. Additionally, as disclosed herein, CTL responses can be primed by conjugating peptides ofthe invention to lipids, such as tripalmitoyl-S- glycerylcysteinlyseryl- serine (P₃CSS). Moreover, an adjuvant such as a synthetic cytosine-phosphorothiolated- guanine-containing (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 the invention, via injection, aerosol, oral, transdermal, transmucosal, intrapleural, infrathecal, or other suitable routes, the immune system ofthe host responds to the vaccine by producing large amounts of CTLs and/or HTLs specific for the desired antigen. Consequently, the host becomes at least partially immune to later development of cells that express or overexpress an antigen of a protein of Figure 2, or the host derives at least some therapeutic benefit when the antigen was tumor- associated. , , 1

In some embodiments, it may be desirable to combine the class I peptide components with components that induce or facilitate neutralizing .antibody and or helper T cell responses directed to the target antigen. A prefeπed embodiment of such a composition comprises class I and class II epitopes in accordance with the invention. An alternative embodiment of such a composition comprises a class I and/or class II epitope in accordance with the invention, along with a cross reactive HTL epitope such as PADRE™ (Epimmune, San Diego, CA) molecule (described e.g., in U.S. Patent Number 5,736,142). A vaccine ofthe invention can also include antigen-presenting cells (APC), such as dendritic cells (DC), as a vehicle to present peptides ofthe 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 prefeπed that each ofthe following principles be balanced in order to make the selection. The multiple epitopes to be incorporated in a given vaccine composition may be, but need not be, contiguous in sequence in the native antigen from which the epitopes are derived.

1.) Epitopes are selected which, upon administration, mimic immune responses that have been observed to be coπelated 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 coπelated 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-peptides, or a sufficient aπay 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.

5.) Of particular relevance are epitopes refeπed 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 ofthe amino terminal epitope and the carboxyl terminus ofthe carboxyl terminal epitope in the peptide. When providing a multi-epitopic sequence, such as a sequence comprising nested epitopes, it is generally important to screen the sequence in order to insure that it does not have pathological or other deleterious biological properties.

6.) If a polyepitopic protein is created, or when creating a minigene, an objective is to generate the smallest peptide that encompasses the epitopes of interest. This principle is similar, if not the same as that employed when selecting a peptide comprising nested epitopes. However, with an artificial polyepitopic peptide, the size minimization objective is balanced against the need to integrate any spacer sequences between epitopes in the polyepitopic protein. Spacer amino acid residues can, for example, be introduced to avoid junctional epitopes (an epitope recognized by the immune system, not present in the target antigen, and only created by the 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 an immune response to that non- native epitope. Of particular concern is a junctional epitope that is a "dominant epitope." A dominant epitope may lead to such a zealous response that immune responses to other epitopes are diminished or suppressed. •

7.) Where the sequences of multiple variants ofthe same target protein are present, potential peptide epitopes can also be selected on the basis of their conservancy. For example, a criterion for conservancy may define that the entire sequence of an HLA class I binding peptide or the entire 9-mer core of a class II binding peptide be conserved in a designated percentage ofthe 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 the peptides ofthe invention are a particularly useful embodiment ofthe invention. Epitopes for inclusion in a minigene are preferably selected according to the guidelines set forth in the previous section. A prefeπed means of administering nucleic acids encoding the peptides ofthe invention uses minigene constructs encoding a peptide comprising one or multiple epitopes ofthe invention.

The use of multi-epitope minigenes is described below and in, Ishioka 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 plasmid encoding supermotif- and/or motif-bearing epitopes derived from a protein ofthe invention, the PADRE® universal helper T cell epitope (or multiple HTL epitopes from a protein ofthe invention), and an endoplasmic reticulum-franslocating signal sequence can be engineered. A vaccine may also comprise epitopes that are derived from other TAAs.

The immunogenicity of a multi-epitopic minigene can be confirmed in transgenic mice to evaluate the magnitude of CTL induction responses against the epitopes tested. Further, the immunogenicity of DNA-encoded epitopes in vivo can be coπelated with the in vitro responses of specific CTL lines against target cells transfected with the DNA plasmid. Thus, these experiments can show that the minigene serves: V.) to generate a CTL response; and, 2.) that the induced CTLs recognized cells expressing the encoded epitopes.

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

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

Standard regulatory sequences well known to those of skill in the art are preferably included in the vector to ensure expression in the target cells. Several vector elements are desirable: a promoter with a down-stream 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., the human cytomegaloviras (hCMV) promoter. See, e.g., U.S. Patent Nos. 5,580,859 and 5,589,466 for other suitable promoter sequences.

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

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

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

In some embodiments, a bi-cisfronic expression vector which allows production of both the minigene- encoded epitopes and a second protein (included to enhance or decrease immunogenicity) can be used. Examples of proteins or polypeptides that could beneficially enhance the immune response if co-expressed include cytokines (e.g., IL-2, IL-12, GM-CSF), cytokine-inducing molecules (e.g., LeIF), costimulatory molecules, or for HTL responses, pan-DR binding proteins (PADRE™, Epimmune, San Diego, CA). Helper (HTL) epitopes can be joined to intracellular targeting signals and expressed separately from expressed CTL epitopes; this allows direction ofthe HTL epitopes to a cell compartment different than that ofthe CTL epitopes. If required, this could facilitate more efficient entry of HTL epitopes into the HLA class II pathway, thereby improving HTL induction. In contrast to HTL or CTL induction, specifically decreasing the immune response by co-expression of immunosuppressive molecules (e.g. TGF-β) may be beneficial in certain diseases.

Therapeutic quantities of plasmid DNA can be produced for example, by fermentation in E. coli, followed by purification. Aliquots from the working cell bank are used to inoculate growth medium, and grown to saturation in shaker flasks or a bioreactor according to well-known techniques. Plasmid 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 reconstitution of lyophilized DNA in sterile phosphate-buffer saline (PBS). This approach, known as "naked DNA," is cuπently being used for intramuscular (IM) administration in clinical trials. To maximize the immunotherapeutic 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, glycohpids, 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 al, Proc. Nat'l Acad. Sci. USA 84:7413 (1987). In addition, peptides and compounds refeπed 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. Electroporation can be used for "naked" DNA, whereas cationic lipids allow direct in vitro transfection. A plasmid expressing green fluorescent protein (GFP) can be co-fransfected to allow enrichment of transfected cells using fluorescence activated cell sorting (FACS). These cells are then chromium-51 (⁵¹Cr) labeled and used as target cells for epitope-specific CTL lines; cytolysis, detected by ^5ICr release, indicates both production of, and HLA presentation of, minigene-encoded CTL epitopes. Expression of HTL epitopes may be evaluated in an analogous manner using assays to assess HTL activity.

In vivo immunogenicity is a second approach for functional testing of minigene DNA formulations. Transgenic mice expressing appropriate human HLA proteins are immunized with the DNA product. The dose and route of administration are formulation dependent (e.g., IM 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, ⁵¹Cr-labeled target cells using standard techniques. Lysis of target cells that were sensitized by HLA loaded with peptide epitopes, coπesponding to minigene-encoded epitopes, demonstrates DNA vaccine function for in vivo induction of CTLs. Immunogenicity of HTL epitopes is confirmed in transgenic mice in an analogous manner.

Alternatively, the nucleic acids can be administered using ballistic delivery as described, for instance, in U.S. Patent No. 5,204,253. Using this technique, particles comprised solely of DNA are administered. In a further alternative embodiment, DNA can be adhered to particles, such as gold particles.

Minigenes can also be delivered using other bacterial or viral delivery systems well known in the art, e.g., an expression construct encoding epitopes ofthe 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 ofthe invention can be modified, e.g., analoged, to provide desired attributes, such as improved serum half life, broadened population coverage or enhanced immunogenicity.

For instance, the ability of a peptide to induce CTL activity can be enhanced by linking the peptide to a sequence which contains at least one epitope that is capable of inducing a T helper cell response. Although a CTL peptide can be directly linked to a T helper peptide, often CTL epitope HTL epitope conjugates are linked by a spacer molecule. The spacer is typically comprised of relatively small, neutral molecules, such as amino acids or amino acid mimetics, which are substantially uncharged under physiological conditions. The spacers are typically selected from, e.g., Ala, Gly, or other neutral spacers of nonpolar amino acids or neutral polar amino acids. It will be understood that the optionally present spacer need not be comprised ofthe same residues and thus may be a hetero- or homo-oligomer. When present, the spacer will usually be at least one or two residues, more usually three to six residues and sometimes 10 or more residues. The CTL peptide epitope can be linked to the T helper peptide epitope either directly or via a spacer either at the amino or carboxy terminus ofthe CTL peptide. The amino terminus of either the immunogenic peptide or the T helper peptide may be acylated. In certain embodiments, the T helper peptide is one that is recognized by T helper cells present in a majority of a genetically diverse population. This can be accomplished by selecting peptides that bind to many, most, or all ofthe 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: ), Plasmodiumfalciparum circumsporozoite (CS) protein at positions 378-398

(DIEK-tOAKMEKASSVFNVVNS; SEQ ID NO: ), and Streptococcus 18kD protein at positions 116-131

(GAVDSILGGVATYGAA; SEQ ID NO: ). Other examples include peptides bearing a DR 1-4-7 supermotif, or either ofthe 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) molecules. For instance, a pan-DR-binding epitope peptide having the formula: aKXVAAWTLKAAa (SEQ ID NO: ), where "X" is either cyclohexylalanine, 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 the 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 can 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 they can be conjugated to other molecules such as lipids, proteins, carbohydrates, and the like to increase their biological activity. For example, a T helper peptide can be conjugated to one or more palmitic 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 ofthe 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 α- 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 an adjuvant, e.g., incomplete Freund's adjuvant. In a prefeπed embodiment, a particularly effective immunogenic composition comprises palmitic acid attached to ε- and α- amino groups of Lys, which is attached via linkage, e.g., Ser-Ser, to the amino terminus ofthe immunogenic peptide.

As another example of lipid priming of CTL responses, E. coli hpoproteins, such as tripalmitoyl-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 ofthe invention can be coupled to P₃CSS, for example, and the lipopeptide adrninistered to an individual to specifically prime an immune response to the target antigen. Moreover, because the induction of neutralizing antibodies can also be primed with P₃CSS- conjugated epitopes, two such compositions can be combined to more effectively elicit both humoral and cell- mediated responses.

X.C.4. Vaccine Compositions Comprising DC Pulsed with CTL and/or HTL Peptides

An embodiment of a vaccine composition in accordance with the invention comprises ex vivo aclministration 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/IL-4. After pulsing the DC with 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 the pulsed peptide epitopes complexed with HLA molecules on their surfaces.

The DC can be pulsed ex vivo with a cocktail of peptides, some of which stimulate CTL responses to a protein of Figure 2. 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 with the invention is used to treat a cancer which expresses or overexpresses a protein of Figure 2.

X.D. Adoptive Immunotherapy

Antigenic peptides ofthe invention, e.g., peptides derived from a protein of Figure 2, are used to elicit a CTL and or HTL response ex vivo, as well. The resulting CTL or HTL cells, can be used to treat tumors in patients that do not respond to other conventional forms of therapy, or will 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 with a source of antigen-presenting cells (APC), such as dendritic cells, and the appropriate immunogenic peptide. After an appropriate incubation time (typically about 7-28 days), in which the precursor cells are activated and expanded into effector cells, the cells are infused back into the patient, where they 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 ofthe invention are typically used to treat and/or prevent a cancer that expresses or overexpresses a Figure 2 protein. 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 the antigen and to cure or at least partially aπest 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., the particular composition aclministered, the manner of aehr-inistration, the stage and severity ofthe disease being treated, the weight and general state of health ofthe patient, and the judgment ofthe prescribing physician. _ .

For pharmaceutical compositions, the immunogenic peptides ofthe invention, or DNA encoding them, are generally administered to an individual already bearing a tumor that expresses a protein of Figure 2. The peptides or DNA encoding them can be administered individually or as fusions of one or more peptide sequences. Patients can be freated with the immunogenic peptides separately or in conjunction with other freatments, such as surgery, as appropriate.

For therapeutic use, administration should generally begin at the first diagnosis of a protein of Figure 2- associated cancer. This is followed by boosting doses until at least symptoms are substantially abated and for a period thereafter. The embodiment ofthe 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 the patient may vary according to the stage ofthe disease or the patient's health status. For example, in a patient with a tumor tiiat expresses a protein of Figure 2, a vaccine comprising CTLs specific for the respective protein of Figure 2 may be more efficacious in killing tumor cells in patient with advanced disease than alternative embodiments. It is generally important to provide an amount ofthe peptide epitope delivered by a mode of adminisfration 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 ofthe invention.

The dosage for an initial therapeutic immunization generally occurs in a unit dosage range where the lower . value is about 1, 5, 50, 500, or 1,000 μg and the higher value is about 10,000; 20,000; 30,000; or 50,000 μg. Dosage values for a human typically range from about 500 μg to about 50,000 μg per 70 kilogram patient. Boosting dosages of between about 1.0 μg to about 50,000 μg of peptide pursuant to a boosting regimen over weeks to months may be administered depending upon the patient's response and condition as determined by measuring the specific activity of CTL and HTL obtained from the patient's blood. Administration should continue until at least clinical symptoms or laboratory tests indicate that the neoplasia, has been eliminated or reduced and for a period thereafter. The dosages, routes of administration, and dose schedules are adjusted in accordance with methodologies known in the art.

In certain embodiments, the peptides and compositions ofthe present invention are employed in serious disease states, that is, life-threatening or potentially life threatening situations. In such cases, as a result ofthe minimal amounts of extraneous substances and the relative nontoxic nature ofthe peptides in prefeπed compositions ofthe invention, it is possible and may be felt desirable by the treating physician to administer substantial excesses of these peptide compositions relative to these stated dosage amounts.

The vaccine compositions ofthe invention can also be used purely as prophylactic agents. Generally the dosage for an initial prophylactic irnmunization generally occurs in a unit dosage range where the lower value is about 1, 5, 50, 500, or 1000 μg and the higher value is about 10,000; 20,000; 30,000; or 50,000 μg. Dosage values for a human typically range from about 500 μg to about 50,000 μg per 70 kilogram patient. This is followed by boosting dosages of between about 1.0 μg to about 50,000 μg of peptide administered at defined intervals from about four weeks to six months after the initial administration of vaccine. The immunogenicity ofthe vaccine can be assessed by measuring the specific activity of CTL and HTL obtained from a sample ofthe patient's blood.

The pharmaceutical compositions for therapeutic treatment are intended for parenteral, topical, oral, nasal, ^■ infrathecal, 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 ofthe 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, the 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 chloride, calcium chloride, sorbitan monolaurate, triethanolamine oleate, etc.

The concenfration of peptides ofthe 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 numan unit dose lorm ot a composition is typically included m a pharmaceutical composition that comprises a human unit dose of an acceptable caπier, in one embodiment an aqueous caπier, 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. 17"' 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 μg, generally 100-5,000 μg, 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 IM (or SC or ID) in the amounts of 0.5-5 mg at multiple sites. The nucleic acid (0.1 to 1000 μg) 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 vims administered at a dose of 5-10⁷ to 5xl0⁹ pfu.

For antibodies, a treatment generally involves repeated administration ofthe antibody preparation, via an acceptable route of administration such as intravenous injection (IV), typically at a dose in the range of about 0.1 to about 10 mg/kg body weight. In general, doses in the range of 10-500 mg mAb per week are effective and well tolerated. Moreover, an initial loading dose of approximately 4 mg/kg patient body weight IV, followed by weekly doses of about 2 mg/kg IV ofthe anti-Figure 2 protein mAb preparation represents an acceptable dosing regimen. As appreciated by those of skill in the art, various factors can influence the ideal dose in a particular case. Such factors include, for example, half life of a composition, the binding affinity of an Ab, the immunogenicity of a substance, the degree of expression ofthe protein ofthe invention in the patient, the extent of circulating shed of protein ofthe invention antigen, the desired steady-state concentration level, frequency of treatment, and the influence of chemotherapeutic or other agents used in combination with the treatment method ofthe invention, as well as the health status of a particular patient. Non-limiting prefeπed human unit doses are, for example, 500μg - lmg, lmg - 50mg, 50mg - lOOmg, lOOmg - 200mg, 200mg - 300mg, 400mg - 500mg, 500mg - 600mg, 600mg - 700mg, 700mg - 800mg, 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, three or four weeks by weekly doses; 225, 250, 275, 300, 325, 350, 375, 400mg m² of body area weekly; l-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 ofthe polynucleotide, molecular weight ofthe polynucleotide and route of administration. Dosages are generally selected by the physician or other health care professional in accordance with a variety of parameters known in the art, such as severity of symptoms, history ofthe patient and the like. Generally, for a polynucleotide of about 20 bases, a dosage range maybe selected from, for example, an independently selected lower limit such as about 0.1, 0.25, 0.5, 1, 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400 or 500 mg/kg up to an independently selected upper limit, greater than the lower limit, of about 60, 80, 100, 200, 300, 400, 500, 750, 1000, 1500, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000 or 10,000 mg/kg. For example, a dose may be about any ofthe 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 a suitable dosage range or effective amount that provides any therapeutic effect. As appreciated by one of ordinary skill in the art, a therapeutic effect depends on a number of factors. Dosages are generally selected by the physician or other health care professional in accordance with a variety of parameters known in the art, such as severity of symptoms, history ofthe patient and the like. A dose may be about 10⁴ cells to about 10⁶ cells, about 10⁶ cells to about 10⁸ cells, about 10⁸ to about 10¹¹ cells, or about 10⁸ 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 10⁸ cells/m² .

Proteins(s) ofthe invention, and/or nucleic acids encoding the protein(s), can also be administered via liposomes, which may also serve to: 1) target the proteins(s) to a particular tissue, such as lymphoid tissue; 2) to target selectively to diseases cells; or, 3) to increase the half-life ofthe peptide composition. Liposomes include emulsions, foams, micelles, insoluble monolayers, liquid crystals, phospholipid dispersions, lamellar layers and the like. In these preparations, the peptide to be delivered is incorporated as part of a liposome, alone or in conjunction with a molecule which binds to a receptor prevalent among lymphoid cells, such as monoclonal antibodies which bind to the CD45 antigen, or with other therapeutic or immunogenic compositions. Thus, liposomes either filled or decorated with a desired peptide ofthe invention can be directed to the site of lymphoid cells, where the liposomes then deliver the peptide compositions. Liposomes for use in accordance with the 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 ofthe liposomes in the blood stream. A, variety of methods are available for preparing liposomes, as described in, e.g., Szoka, et al, Ann. Rev. Biophys. Bioeng. 9:467 (1980), and U.S. Patent Nos. 4,235,871, 4,501,728, 4,837,028, and 5,019,369.

For targeting cells ofthe immune system, a ligand to be incorporated into the liposome can include, e.g., . antibodies or fragments thereof specific for cell surface determinants ofthe 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, the manner of administration, the peptide being delivered, and the stage ofthe disease being freated.

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 the like. For oral administration, a pharmaceutically acceptable nontoxic composition is formed by incorporating any ofthe normally employed excipients, such as those caπiers previously listed, and generally 10-95% of active ingredient, that is, one or more peptides ofthe invention, and more preferably at a concentration of 25%-75%.

For aerosol aclministratiόn, immunogenic peptides are preferably supplied in finely divided form along with a surfactant and propellant. Typical percentages of peptides are about 0.01%-20% by weight, preferably about 1%-10%. The surfactant must, of course, be nontoxic, and preferably soluble in the propellant. Representative of such agents are the esters or partial esters of fatty 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 0.1%-20% by weight ofthe composition, preferably about 0.25-5%. The balance ofthe composition is ordinarily propellant. A carrier can also be included, as desired, as with, e.g., lecithin for intranasal delivery.

XI.) Diagnostic and Prognostic Embodiments of the Invention.

As disclosed herein, polynucleotides, polypeptides, reactive cytotoxic T cells (CTL), reactive helper T cells (HTL) and anti-polypeptide antibodies ofthe invention 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 Example 4).

Proteins of Figure 2 can be analogized to the 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., Meπill et al, 3. 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 Figure 2 polynucleotides and polypeptides (as well as Figure 2-related polynucleotide probes and anti- Figure 2 protein antibodies used to identify the presence of these molecules) and their properties allows skilled artisans to utilize these molecules in methods that are analogous to those used, for example, in a variety of diagnostic assays directed to examining conditions associated with cancer.

Typical embodiments of diagnostic methods, which utilize the polynucleotides, polypeptides, reactive T cells and antibodies ofthe invention, are analogous to those methods 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 Figure 2 polynucleotides described herein can be utilized in the same way to detect the respective Figure 2 protein overexpression or the metastasis of prostate and other 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 methods 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., Alanen et al, Pathol. Res. Pract. 192(3):233-7 (1996)), the Figure 2 polypeptides described herein can be utilized to generate antibodies for use in detecting the respective proteins of Figure 2 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 the lung or prostate gland etc.) to a different area ofthe body (such as a lymph node), assays which examine a biological sample for the presence of cells expressing Figure 2 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 gene or protein of Figure 2-expressing cells (e.g., a lymph node) is found to contain a protein of Figure 2-expressing cells, this finding is indicative of metastasis.

Alternatively polynucleotides and/or polypeptides ofthe invention can be used to provide evidence of cancer, for example, when cells in a biological sample that do not normally express Figure 2 genes or express Figure 2 genes at a different level are found to express Figure 2 genes or have an increased expression of Figure 2 genes (see, e.g., the expression in the cancers of tissues listed in Table I and in patient samples etc. shown in the accompanying Figures). In such assays, artisans may further wish to generate supplementary evidence of metastasis by testing the biological sample for the presence of a second tissue restricted marker (in addition to a protein of Figure 2) 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, a gene of Figure 2 polynucleotide fragments and polynucleotide variants are used in an analogous manner. In particular, typical PSA polynucleotides used in methods of monitoring PSA are probes or primers which consist of fragments ofthe PSA cDNA sequence. Illustrating this, primers used to PCR amplify a PSA polynucleotide must include less than the whole PSA sequence to function in the polymerase chain reaction. In the context of such PCR reactions, skilled artisans generally create a variety of different 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 al, Methods Mol. Biol. 98: 121-154 (1998)). An additional illustration ofthe use of such fragments is ' provided in Example 4, where a gene of Figure 2 polynucleotide fragments are used as a probe to show the expression of respective gene of Figure 2 RNAs in cancer cells. In addition, variant polynucleotide sequences are typically used as primers and probes for the coπesponding mRNAs in PCR and Northern analyses (see, e.g., Sawai et al, Fetal Diagn. Ther. 1996 Nov-Dec 11(6):407-13 and Cuπent 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 are capable of binding to a target polynucleotide sequence (e.g., a Figure 2 polynucleotide or variant thereof) under conditions of high stringency.

Furthermore, PSA polypeptides which contain an epitope that can be recognized by an antibody or T cell - > that specifically binds to that epitope are used in methods of monitoring PSA. Polypeptide fragments, polypeptide analogs or variants of a protein of Figure 2 can also be used in an analogous manner. This practice of using polypeptide fragments or polypeptide variants to generate antibodies (such as anti-PSA antibodies or T cells) is typical in the art with a wide variety of systems such as fusion proteins being used by practitioners (see, e.g., Cuπent Protocols In Molecular Biology, Volume 2, Unit 16, Frederick M. Ausubel et al. eds., 1995). In this context, each epitope(s) functions to provide the architecture with which an antibody or T cell is reactive. Typically, skilled artisans create a variety of different polypeptide fragments that can be used in order to generate immune responses specific for different portions of a polypeptide of interest (see, 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 ofthe biological motifs of a protein of Figure 2 discussed herein or a motif-bearing subsequence which is readily identified by one of skill in the art based on motifs available in the art. Polypeptide fragments, variants or analogs are typically useful in this 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 protein of Figure 2).

As shown herein, the Figure 2 polynucleotides and polypeptides (as well as the Figure 2 polynucleotide probes and anti-proteins of Figure 2 antibodies or T cells used to identify the presence of these molecules) exhibit specific properties that make them useful in diagnosing cancers such as those listed in Table I. Diagnostic assays that measure the presence of gene of Figure 2 gene products, in order to evaluate the presence or onset of a disease condition described herein, such as prostate cancer, are used to identify patients for preventive measures or further monitoring, as has been done so 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 al., Pathol. Res. Pract. 192(3): 233-237 (1996)), and consequently, materials such as Figure 2 polynucleotides and polypeptides (as well as the gene of Figure 2 polynucleotide probes and anti-proteins of Figure 2 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 Figure 2 polynucleotides disclosed herein have a number of other utilities such as their use in the identification of oncogenetic associated chromosomal abnormalities in the chromosomal region to which a Figure 2 gene maps (see Example 3 below). Moreover, in addition to their use in diagnostic assays, the Figure 2-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 Int 1996 Jun 28_.80(1-2): 63-9).

Additionally, Figure 2-related proteins or polynucleotides ofthe invention can be used to treat a pathologic condition characterized by the over-expression of Figure 2 proteins. For example, the amino acid or nucleic acid sequences in Figure 2 or Figure 3, or fragments of either, can be used to generate an immune response to a protein of Figure 2 antigen. Antibodies or other molecules that react with proteins ofthe invention Figure 2 can be used to modulate the function of this molecule, and thereby provide a therapeutic benefit.

Xπ.) Inhibition of the Function of a Protein in the Invention

The invention includes various methods and compositions for inhibiting the binding of proteins in Figure 2 to its binding partner or its association with other protein(s) as well as methods for inhibiting the function of proteins in Figure 2.

Xπ.A.) Inhibition of a Protein of Figure 2 with Intracellular Antibodies

In one approach, a recombinant vector that encodes single chain antibodies that specifically bind to a Figure 2 protein are introduced into proteins of Figure 2 expressing cells via gene transfer technologies. Accordingly, the encoded single chain anti-protein of Figure 2 antibodies are expressed intracellularly, and bind to the respective Figure 2 protein, and thereby inhibits its function. Methods for engineering such intracellular single chain antibodies are well known. Such intracellular antibodies, also known as "infrabodies", are specifically targeted to a particular compartment within the cell, providing control over where the inhibitory activity ofthe treatment is focused. This technology has been successfully applied in the art (for review, see Richardson and Marasco, 1995, TIBTECH vol. 13). Infrabodies have been shown to virtually eliminate the expression of otherwise abundant cell surface receptors (see, e.g., Richardson et al, 1995, Proc. Natl. Acad. Sci. USA 92: 3137-3141; Beerli et al, 1994, J. Biol. Chem. 289: 23931-23936; Deshane et al, 1994, Gene Ther. 1: 332-337).

Single chain antibodies comprise the variable domains ofthe 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 chain antibodies in order to precisely target the intrabody to the desired intracellular compartment. For example, infrabodies targeted to the 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. Infrabodies intended to exert activity in the nucleus are engineered to include a nuclear localization signal. Lipid moieties are joined to infrabodies in order to tether the intrabody to the cytosolic side ofthe plasma membrane. Infrabodies can also be targeted to exert function in the cytosol. For example, cytosolic infrabodies are used to sequester factors within the cytosol, thereby preventing them from being transported to their natural cellular destination. In one embodiment, infrabodies are used to capture proteins of Figure 2 in the nucleus, thereby preventing the activity of that protein(s) within the nucleus. Nuclear targeting signals are engineered into such Figure 2-related infrabodies in order to achieve the desired targeting. Such Figure 2-related infrabodies are designed to bind specifically to a particular Figure 2 protein domain. In another embodiment, cytosolic infrabodies that specifically bind to a Figure 2 protein are used to prevent the protein in Figure 2 from gaining access to the nucleus, thereby preventing it from exerting any biological activity within the nucleus (e.g., preventing proteins of Figure 2 from forming transcription complexes with other factors).

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

XII.B.) Inhibition of a Protein of Figure 2 with Recombinant Proteins

In another approach, recombinant molecules bind to a Figure 2 protein and thereby inhibit the function of a protein of Figure 2. For example, these recombinant molecules prevent or inhibit Figure 2 proteins from accessing/binding to its binding partner(s) or associating with other protein(s). Such recombinant molecules can, for ; example, contain the reactive part(s) of an antibody molecule specific for a protein of Figure 2. hi a particular embodiment, the Figure 2 protein binding domain of a coπesponding binding partner is engineered into a dimeric fusion protein, whereby the fusion protein comprises two protein of Figure 2 ligand binding domains linked to the Fc portion of a human IgG, such as human IgGl . Such IgG portion can contain, for example, the C_H2 and C_H3 domains and the hinge region, but not the C 1 domain. Such dimeric fusion proteins are aclministered in soluble form to patients suffering from a cancer associated with the expression of proteins ofthe invention, see, e.g., Figure 2, whereby the dimeric fusion protein specifically binds to a Figure 2 protein and blocks the interaction of a Figure 2 protein with one or more binding partners. Such dimeric fusion proteins are further combined into multimeric proteins using known antibody linking technologies.

XH.C.) Inhibition of Transcription or Translation in Accordance with the Invention

The present invention also comprises various methods and compositions for inhibiting the transcription of a Figure 2 gene. Similarly, the invention also provides methods and compositions for inhibiting the translation of the genes in Figure 2-related mRNA into protein.

In one approach, a method of inhibiting the franscription of a Figure 2 gene comprises contacting the Figure 2 gene with a respective Figure 2 antisense polynucleotide. In another approach, a method of inhibiting gene of Figure 2-related mRNA translation comprises contacting a gene of Figure 2-related mRNA with an antisense polynucleotide. In another approach, agene of Figure 2 specific ribozyme is used to cleave a gene of Figure 2- relatedmessage, thereby inhibiting translation. Such antisense and ribozyme based methods can also be directed to the regulatory regions of a Figure 2 gene, such as a promoter and/or enhancer element for a gene of Figure 2. Similarly, proteins capable of inhibiting agene of Figure 2 transcription factor are used to inhibit the gene of Figure 2 mRNA franscription. 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.

Other factors that inhibit the transcription of a Figure 2 gene by interfering with that gene's franscriptional activation are also useful to treat cancers expressing genes of Figure 2. Similarly, factors that interfere with a gene of Figure 2 gene processing are useful to freat cancers that express genes of Figure 2. Cancer treatment methods utilizing such factors are also within the scope ofthe invention. XII.D.) General Considerations for Therapeutic Strategies

Gene transfer and gene therapy technologies can be used to deliver therapeutic polynucleotide molecules to tumor cells synthesizing proteins ofthe invention, see, e.g., Figure 2, (e.g., antisense, ribozyme, polynucleotides encoding infrabodies and other gene/protein of Figure 2 inhibitory molecules). A number of gene therapy approaches are known in the art. Recombinant vectors encoding Figure 2 antisense polynucleotides, ribozymes, factors capable of interfering with transcription of a gene of Figure 2, 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 ofthe invention can enable the use of reduced dosages of chemotherapy (or other therapies) and/or less frequent achninistration, an advantage for all patients and particularly for those that do not tolerate the toxicity ofthe 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 which a therapeutic composition will inhibit the binding of a protein of Figure 2 to one or more of its binding partners, etc.

In vivo, the effects of a therapeutic composition ofthe invention can be evaluated in a suitable animal model. For example, xenogenic prostate cancer models can be used, wherein human prostate cancer explants or passaged xenograft tissues are introduced into immune compromised animals, such as nude or SCID mice (Klein et al, 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 the formation of osteoblastic metastases characteristic of late stage disease. Efficacy can be predicted using assays that measure inhibition of tumor formation, tumor regression or metastasis, and the like.

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

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

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

Dosages and administration protocols for the treatment of cancers using the foregoing methods will vary with the method and the target cancer, and will generally depend on a number of other factors appreciated in the art.

XIII.) Kits

For use in the diagnostic and therapeutic applications described herein, kits are also within the scope ofthe invention. Such kits can comprise a carrier, package or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each ofthe container(s) comprising one ofthe separate elements to be used in the method. For example, the container(s) can comprise a probe that is or can be detectably labeled. Such probe can be an antibody or polynucleotide specific for a Figure 2-related protein or a Figure 2 gene or message, respectively. Where the method utilizes nucleic acid hybridization to detect the target nucleic acid, the kit can also have containers containing nucleotide(s) for amplification ofthe 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 ofthe amino acid sequences in Figure 2 or Figure 3 or analogs thereof, or a nucleic acid molecules that encodes such amino acid sequences.

The kit ofthe invention will typically comprise the container described above and one or more other containers comprising materials desirable from a commercial and user standpoint, including buffers, diluents, filters, needles, syringes; carrier, package, container, vial and/or tube labels listing contents and/or instructions for use, and package inserts with instructions for use.

A label can be present on the container to indicate that the 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 the kit.

EXAMPLES:

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

Example 1: SSH-Generated Isolation of a cDNA Fragment of the target of the invention Gene

The suppression subtractive hybridization (SSH) cDNA fragments shown in Figure 1 were derived from many different subtractions utilizing LAPC xenografts in differing states of androgen dependence and or casfration as well as using cancer patient derived tissues. The cancer patient tissue SSHs utilized prostate, bladder, and kidney with tumors representing all stages and grades ofthe diseases. Information for additional sequences disclosed in Figure 2 and Figure 3 were derived from other clones and the use of various sequence databases.

Materials and Methods

LAPC Xenografts and Human Tissues:

LAPC xenografts were obtained from Dr. Charles Sawyers (UCLA) and generated as described (Klein et al, 1997, Nature Med. 3: 402-408; Craft et al., 1999, Cancer Res. 59: 5030-5036). Androgen dependent and independent LAPC xenografts were grown in male SCID mice and were passaged as small tissue chunks in recipient males. LAPC xenografts were derived from LAPC tumors. To generate the androgen independent (Al) xenografts, male mice bearing androgen dependent (AD) tumors were casfrated and maintained for 2-3 months. After the tumors re- grew, the tumors were harvested and passaged in castrated males or in female SCID mice. Tissues from prostate, bladder, kidney, colon, lung, pancreas, ovary and breast cancer patients as well as the coπesponding normal tissues were stored frozen at -70 C prior to RNA isolation.

RNA Isolation:

Tumor tissue and cell lines were homogenized in Trizol reagent (Life Technologies, Gibco BRL) using 10 ml/ g tissue or 10 ml/ 10⁸ cells to isolate total RNA. Poly A RNA was purified from total RNA using Qiagen's Oligotex mRNA Mini and Midi kits. Total and mRNA were quantified by spectrophotomettic analysis (O.D. 260/280 nm) and analyzed by gel elecfrophoresis.

Oligonucleotides:

The following HPLC purified oligonucleotides were used.

DPNCDN (cDNA synthesis primer):

5 'TTTTGATCAAGCTT₃_3 ' (SEQ ID NO: XX)

Adaptor 1 :

5'CTAATACGACTCACTATAGGGCTCGAGCGGCCGCCCGGGCAG3' (SEQ ID NO: XX)

3'GGCCCGTCCTAG5' (SEQ ID NO: XX) Adaptor 2: 5'GTAATACGACTCACTATAGGGCAGCGTGGTCGCGGCCGAG3' (SEQ ID NO:XX)

3'CGGCTCCTAG5' (SEQ ID NO: XX) PCR primer 1: 5 'CTAATACGACTCACTATAGGGC3 ' (SEQ ID NO: XX)

Nested primer (NP) 1 :

5 'TCGAGCGGCCGCCCGGGCAGGA3 ' (SEQ ID NO: XX)

Nested primer fNP)2:

5 ' AGCGTGGTCGCGGCCGAGGA3 ' (SEQ ID NO: XX)

Suppression Subtractive Hybridization:

Suppression Subtractive Hybridization (SSH) was used to identify cDNAs coπesponding to genes that are differentially expressed in cancer. The SSH reaction utilized cDNA from the prostate cancer xenografts, LAPC-4 AD, LAPC-4 Al, LAPC-9 AD, and LAPC-9AI as well as from prostate, bladder, and kidney cancer patients. Specifically, to isolate genes that are involved in the progression of androgen dependent (AD) prostate cancer to androgen independent (Al) cancer, experiments were conducted with the LAPC-9 AD and LAPC-4 AD xenograft in male SCID mice. Mice that harbored these xenografts were castrated when the tumors reached a size of 1 cm in diameter. The tumors regressed in size and temporarily stopped producing the androgen dependent protein PSA. Seven to fourteen days post-castration, PSA levels were detectable again in the blood ofthe mice. Eventually the tumors develop an Al phenotype and start growing again in the casfrated males. Tumors were harvested at different time points after castration to identify genes that are turned on or off during the transition to androgen independence.

The cDNAs derived from LAPC-4 AD and LAPC-9 AD tumors (post-castration) were used as the source ofthe "tester" cDNAs, while the cDNAs from LAPC4-AD and LAPC-9 AD tumors (grown in intact male mouse) were used as the source ofthe "driver" cDNAs respectively. Some SSHs also used any combination ofthe LAPC-4 AD, LAPC-4 Al, LAPC-9 AD, and LAPC9-AI xenografts as "tester" or "driver". In addition, cDNAs derived from patient tumors of prostate, bladder and kidney cancer were used as "tester" while cDNAs derived from normal prostate, bladder, and kidney were used as "driver" respectively. Double stranded cDNAs coπesponding to tester and driver cDNAs were synthesized from 2 μg of ρoly(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. PTI 117-1, Catalog No. K1804-1). The resulting cDNA was digested with Dpn II for 3 hrs at 37°C. Digested cDNA was extracted with phenol/chloroform (1:1) and ethanol precipitated.

Tester cDNA was generated by diluting 1 μl of Dpn II digested cDNA from the relevant xenograft source (see above) (400 ng) in 5 μl of water. The diluted cDNA (2 μl, 160 ng) was then ligated to 2 μl of Adaptor 1 and Adaptor 2 (10 μM), in separate ligation reactions, in a total volume of 10 μl at 16°C overnight, using 400 u of T4 DNA ligase (CLONTECH). Ligation was terminated with 1 μl of 0.2 M EDTA and heating at 72°C for 5 min.

The first hybridization was performed by adding 1.5 μl (600 ng) of driver cDNA to each of two tubes containing 1.5 μl (20 ng) Adaptor 1- and Adaptor 2- ligated tester cDNA. In a final volume of 4 μl, 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 μl of fresh denatured driver cDNA and were allowed to hybridize overnight at 68°C. The second hybridization was then diluted in 200 μl of 20 mM Hepes, pH 8.3, 50 mM NaCl, 0.2 mM 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 the primary PCR reaction 1 μl ofthe diluted final hybridization mix was added to 1 μl of PCR primer 1 (10 μM), 0.5 μl dNTP mix (10 μM), 2.5 μl 10 x reaction buffer (CLONTECH) and 0.5 μl 50 x Advantage cDNA polymerase Mix (CLONTECH) in a final volume of 25 μl. PCR 1 was conducted using the following conditions: 75°C for 5 min., 94^CC 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 μl 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 μM) 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 72oC 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 aπayed into 96 well plates and were grown in liquid culture overnight. To identify inserts, PCR amplification was performed on 1 μl of bacterial culture using the conditions of PCRl and NPl and 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 ofthe GenBank, dBest, and NCI-CGAP databases.

A full-length cDNA clone can be identified by assembling EST fragments homologous to the SSH fragment into a large contiguous sequence with an ORF and amplifying the ORF by PCR using xenograft, prostate, bladder, kidney, prostate cancer, bladder cancer, or kidney cancer first strand cDNA.

Example 2: Full Length Cloning of a Target ofthe Invention '

Full length cDNA clones were isolated by a variety of methods known in the art. For example, cDNA phage libraries were constructed from normal and cancer tissues using methods based on those set forth in Cuπent Protocols in Molecular Biology, Ed Ausubel et al., page 5.01. to 5.11.1, through supplement 52, Wiley and Sons; Molecular Cloning, 2^nd Edition, Sambrook et al. Eds, pp. 8.2 to 8.45, 1989, Cold Spring Harbor Press) and full length cDNA clone isolated using probes derived from SSH clones and methods based on (Ausubel et al., supra, pp. 6.0.1 to 6.5.2; Sambrook et al. Eds, supra, 1989, pp. 8.46 to 8.86). In addition, some full length cDNAs were cloned using PCR with primers derived from the extreme ends of ORFs identified in ESTs assembled into contigs. The PCR product is subsequently cloned into pCR2.1. cloning vector (Invitrogen, Carlsbad, CA). Sequences ofthe cloned genes are listed in Figure 2.

Example 3: Chromosomal Mapping

Chromosomal localization can implicate genes in disease pathogenesis. 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, Huntsville Al), human-rodent somatic cell hybrid panels such as is available from the Coriell Institute (Camden, New Jersey), and genomic viewers utilizing BLAST homologies to sequenced and mapped genomic clones (NCBI, Bethesda, Maryland).

Using Figure 2 gene sequences and the NCBI BLAST tool: (see World Wide Web URL www.ncbi.nlm.nih.gov/genome/seq/page.cgi?F=HsBlast.html&&ORG=Hs). placed the genes of Figure 2 to the chromosome locations listed in Table XXII.

Accordingly, as the human genes set forth in Figure 2 map to the designated chromosomes, polynucleotides encoding different regions ofthe of Figure 2 protein can be used to characterize cytogenetic abnormalities on a respective chromosome For example, when chromosomal abnormalities in a chromosome listed in Table XXII have been identified as frequent cytogenetic abnormalities in different cancers (see, e.g., Lai et al., 2000, Clin. Cancer Res. 6(8):3172-6; Oya and Schulz, 2000, Br. J. Cancer 83(5):626-31; Svaren et al., Sept. 12, 2000, J. Biol. Chem.); polynucleotides encoding specific regions ofthe of a Figure 2 protein provide new tools that are used to delineate, with greater precision than previously possible, the specific nature ofthe cytogenetic abnormalities in this region ofthe respective chromosome that contribute to the malignant phenotype. In this context, these polynucleotides satisfy a need in the art for expanding the sensitivity of chromosomal screening in order to identify more subtle and less common chromosomal abnormalities (see, e.g., Evans et al, 1994, Am. J. Obstet. Gynecol. 171(4):1055-1057).

Example 4: Expression Analysis of a Gene of the Invention in Normal Tissues and Patient Specimens

Expression analysis by RT-PCR and Northern analysis demonstrated that normal tissue expression of a gene of Figure 2 is restricted predominantly to the tissues set forth in Table I.

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

RT-PCR analysis and Northern blotting were used to evaluate gene expression in a selection of normal and cancerous urological tissues. The results are summarized in Figures 15-74.

RT-PCR Expression Analysis:

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

Normalization ofthe first sfrand cDNAs from multiple tissues was performed by using the primers 5'atatcgccgcgctcgtcgtcgacaa3' (SEQ ID NO: XX) and 5'agccacacgcagctcattgtagaagg 3' (SEQ ID NO: XX) to amplify β-actin. First strand cDNA (5 μl) were amplified in a total volume of 50 μl containing 0.4 μM primers, 0.2 μM each dNTPs, IXPCR buffer (Clontech, 10 mM Tris-HCL, 1.5 mM MgCl₂, 50 mM KCl, pH8.3) and IX Klentaq DNA polymerase (Clontech). Five μl ofthe PCR reaction can be removed at 18, 20, and 22 cycles and used for agarose gel elecfrophoresis. 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, the band intensities ofthe 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. Three rounds of normalization can be required to achieve equal band intensities in all tissues after 22 cycles of PCR.

To determine expression levels ofthe gene, 5 μl of normalized first strand cDNA are 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. RT-PCR expression analysis is performed on first strand cDNAs generated using pools of tissues from multiple samples. The cDNA normalization was demonstrated in every experiment using beta-actin PCR. Northern Blot Expression Analysis:

Expression of mRNA in normal and cancerous human tissues was analyzed by northern blotting. Expression in normal tissues was analyzed using two multiple tissue blots (Clontech; Palo Alto, California), comprising a total of 16 different normal human tissues, using labeled SSH fragment as a probe. To further analyze expression in prostate cancer tissues, northern blotting was performed on RNA derived from the LAPC xenografts and/or prostate cancer patient samples. In addition, expression in other cancers was studied using patient samples and/or various cancer cell lines.

Figure 15 shows expression of 74P3B3 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), two prostate metastasis to lymph node (LN) harvested from two different patients, prostate cancer pool, bladder cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 74P3B3, was performed at 26 and 30 cycles of amplification. Results show sfrong expression of 74P3B3 in the two prostate metastasis to LN specimens and in prostate cancer pool. Expression was also detected in bladder cancer pool, cancer metastasis pool, and vital pool 2 but not in the vital pool 1.

Figure 16 shows expression of 74P3B3 in normal tissues. Two multiple tissue northern blots (A and B; Clontech) both with 2 μg of mRNA/lane, and a LAPC xenograft blot with 10 μg of total RNA/lane (C) were probed with the 74P3B3 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of approximately 7 kb 74P3B3 transcript in prostate but not in the other normal tissues tested. Expression was also detected in LAPC-4AD and LAPC-4AI but not in LAPC-9AD and LAPC-9AI.

Figure 17 shows expression of 74P3B3 in prostate cancer patient specimens. RNA was extracted from normal prostate (NP), pool of 3 prostate cancer patient tumors (T) and their normal adjacent tissues (N). Northern blot with 10 mg of total RNA/lane was probed with 74P3B3 SSH sequence. Size standards in kilobases (kb) are indicated on the side. The results show strong expression of 74P3B3 in normal prostate and in patient prostate cancer specimens.

Figure 18 shows expression of 74P3B3 in patient cancer specimens. Expression of 74P3B3 was assayed in a panel of human cancers (T) and their respective matched normal tissues (N) on RNA dot blots. Upregulated expression of 74P3B3 in tumors compared to normal tissues was observed in prostate, kidney, breast and colon tumors. The expression detected in normal adjacent tissues (isolated from diseased tissues) but not in normal tissues (isolated from healthy donors) may indicate that these tissues are not fully normal and that 74P3B3 may be expressed in early stage tumors.

Figure 19 shows expression of 83P4B8 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 83P4B8, was performed at 30 cycles of amplification. Results show strong expression of 83P4B8 in all cancer pools tested. Very low expression was detected in the vital pools.

Figure 20 shows expression of 83P4B8 in normal tissues. Two multiple tissue northern blots (A and B; Clontech) both with 2 μg of mRNA/lane, and a LAPC xenograft blot with 10 μg of total RNA/lane (C) were probed with the 83P4B8 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of two approximately 4.4kb 83P4B8 transcripts in testis and to lower level in thymus but not in the other normal tissues tested. Expression was also detected in all 4 LAPC prostate cancer xenografts. Figure 21 shows expression of 83P4B8 in patient cancer specimens and normal tissues. RNA was extracted from a pool of three prostate cancers (PC), bladder cancers (BC), kidney cancers (KC), colon cancers (CC), lung cancers (LC), ovary cancers (OC), cancer metastasis (Met), pancreas cancers (PaC), as well as from normal prostate (NP), normal bladder (NB), normal kidney (NK), normal colon (NC), normal lung (NL), normal breast (NBr) normal ovary (NO) and normal pancreas (NPa). Northern blot with 10 mg of total RNA lane was probed with 83P4B8 sequence. Size standards in kilobases (kb) are indicated on the side. Results show expression of 83P4B8 in the bladder cancers and ovary cancers. Expression of 83P4B8 was also detected in prostate cancers, kidney cancers, colon cancers, lung cancers, cancer metastasis and pancreas cancer but not in the normal tissues tested.

Figure 22 shows expression of 83P4B8 in prostate cancer patient specimens. RNA was extracted from normal prostate (NP), prostate cancer patient tumors (T) and their normal adjacent tissues (N). Northern blot with 10 mg of total RNA/lane was probed with 83P4B8 SSH sequence. Size standards in kilobases (kb) are indicated on the side. The results show sfrong expression of 83P4B8 in the patient prostate cancer specimens.

Figure 23 shows expression of 83P4B8 in colon cancer patient specimens. RNA was extracted from colon cancer cell lines (CL), normal colon (N), colon cancer patient tumors (T) and their normal adjacent tissues (Nat). Northern blots with 10 μg of total RNA were probed with the 83P4B8 SSH fragment. Size standards in kilobases are indicated on the side. Results show strong expression of 83P4B8 in the colon tumor tissues and in all three colon cancer cell lines tested, but not in the normal tissues.

Figure 24 shows expression of 109P1D4 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool, and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi- quantitative PCR, using primers to 109P1D4, was performed at 30 cycles of amplification. Results show strong expression of 109P1D4 in all cancer pools tested. Very low expression was detected in the vital pools

Figure 25 shows expression of 109P1D4 in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA/lane, were probed with the 109P1D4 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of approximately 10 kb 109P1D4 transcript in ovary. Weak expression was also detected in placenta and brain, but not in the other normal tissues tested.

Figure 26 shows expression of 109P1D4 in human cancer cell lines. RNA was extracted from a number of human prostate and bone cancer cell lines. Northern blots with 10 μg of total RNA lane were probed with the 109P1D4 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of 109P1D4 in LAPC-9AD, LAPC-9 Al, LNCaP prostate cancer cell lines, and in the bone cancer cell lines, SK-ES-1 and RD-ES.

Figure 27 shows expression of 109P1D4 ^m patient cancer specimens. Expression of 109P1D4 was assayed in a panel of human cancers (T) and their respective matched normal tissues (N) on RNA dot blots. Upregulated expression of 109P1D4 in tumors compared to normal tissues was observed in uterus, lung and stomach. The expression detected in normal adjacent tissues (isolated from diseased tissues) but not in normal tissues (isolated from healthy donors) may indicate that these tissues are not fully normal and that 109P1D4 may be expressed in early stage tumors.

Figure 28 shows expression of 151P1C7A by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC prostate cancer xenograft pool (LAPC- 4AD, LAPC-4AI, LAPC-9 AD and LAPC-9 Al), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 151P1C7A, was performed at 26 and 30 cycles of amplification. Results show strong expression of 15 IP 1 C7A in bladder, lung, and metastasis cancer pools tested. Expression was also detected in xenograft, prostate, kidney and colon cancer pools but not in the vital pools.

Figure 29 shows expression of 151P1C7A in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA lane, were probed with the 151P1C7A SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of an approximately 2 kb 151P1C7A franscript in placenta but not in the other normal tissues tested.

Figure 30 shows expression of 151P1C7A in bladder cancer patient specimens. RNA was extracted from bladder cancer cell lines (CL; UM-UC-3, J82, SCaBER), normal bladder (Nb), bladder cancer patient tumors (T) and their normal adjacent tissue (N) isolated from bladder cancer patients. Northern blots with 10 μg of total RNA were probed with the 151P1C7A SSH sequence. Size standards in kilobases are indicated on the side. Results show expression of 151P1C7A in patient bladder cancer tissues, and in all bladder cancer cell lines tested, but not in normal bladder.

Figure 31 shows expression of 151P1C7A in prostate cancer patient specimens. RNA was extracted from normal prostate (NP), prostate cancer patient tumors (T) and their normal adjacent tissues (N). Northern blot with 10 mg of total RNA/lane was probed with 151P1C7A SSH sequence. Size standards in kilobases (kb) are indicated on the side. Results show expression of 151 P 1 C7A in the patient prostate cancer specimens.

Figure 32 shows expression of 151P4E11 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC prostate cancer xenograft pool (LAPC- 4AD, LAPC-4AI, LAPC-9 AD and LAPC-9 Al), prostate cancer pool, bladder cancer pool, colon cancer pool, lung cancer pool, ovary 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 151P4E11, was performed at 26 and 30 cycles of amplification. Results show sfrong expression of 151P4E11 in all cancer pools tested. Expression was detected in vital pool 2 but not in vital pool 1.

Figure 33 shows expression of 151P4E11 in normal tissues. Two multiple tissue northern blots (A and B; Clontech) both with 2 μg of mRNA/lane, and a LAPC xenograft blot with 10 μg of total RNA lane (C) were probed with the 151P4E11 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of an approximately 1.2 kb 151P4E11 transcript in prostate, testis, colon and small intestine. Expression was also detected in all the LAPC prostate cancer xenografts LAPC-4AD, LAPC-4AI, and LAPC-9 Al, but not in LAPC-9AD.

Figure 34 shows expression of 154P2A8 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 154P2A8, was performed at 26 and 30 cycles of amplification. Results show strong expression of 154P2A8 in bladder cancer pool and lung cancer pool. Expression was also detected in prostate cancer pool, kidney cancer pool, colon cancer pool, and cancer metastasis pool but not in vital pool 1 and vital pool 2.

Figure 35 shows expression of 156P1D4 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC prostate cancer xenograft pool (LAPC- 4AD, LAPC-4AI, LAPC-9 AD and LAPC-9 Al), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary 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 156P1D4, was performed at 26 and 30 cycles of amplification. Results show strong expression of 156P1D4 in kidney cancer pool and vital pool 1. Expression was also detected in xenograft pool, prostate cancer pool, bladder cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool and vital pool 2.

Figure 36 shows expression of 156P1D4 in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA/lane, were probed with the 156P1D4 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of an approximately 2 kb 156P1D4 transcript in kidney and prostate but not in the other normal tissues tested.

Figure 37 shows expression of 156P1D4 in kidney cancer patient specimens. RNA was extracted from normal kidney (Nk), kidney cancer patient tumors (T) and their normal adjacent tissues (N). Northern blots with 10 μg of total RNA were probed with the 156P1D4 SSH fragment. Size standards in kilobases are indicated on the side. Results show strong expression of 156P1D4 in all kidney tumor tissues tested. The expression of 156P1D4 detected in tumor tissues is stronger than in normal tissues.

Figure 38 shows expression of 156P5C12 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC prostate cancer xenograft pool (LAPC- 4AD, LAPC-4AI, LAPC-9AD and LAPC-9 AT), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary 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 156P5C12, was performed at 26 and 30 cycles of amplification. Results show strong expression of 156P5C12 in kidney cancer pool and vital pool 1. Expression was also detected in xenograft pool, prostate cancer pool, bladder cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool and vital pool 2.

Figure 39 shows expression of 156P5C12 in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA lane, were probed with the 156P5C12 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of an approximately 1.4 kb 156P5C12 transcript in kidney but not in the other normal tissues tested.

Figure 40 shows expression of 156P5C12 in kidney cancer patient specimens. RNA was extracted from kidney cancer cell lines (CL; 769-P, A498, SW839), normal kidney (N), kidney cancer patient tumors (T) and their normal adjacent tissues (NAT).' Northern blots with 10 μg of total RNA were probed with the 156P5C12 SSH fragment. Size standards in kilobases are indicated on the side. Results show expression of 156P5C12 in normal tissues, and in some but not all kidney tumor tissues. Expression was absent in the kidney cancer cell lines tested.

Figure 41 shows expression of 159P2B5 by RT-PCR. First sfrand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), and bladder cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 159P2B5, was performed at 26 and 30 cycles of amplification. Results show expression of 159P2B5 in bladder cancer pool tested but not in the vital pools.

Figure 42 shows expression of 159P2B5 in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA lane, were probed with the 159P2B5 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show very weak expression of an approximately 4.5 kbl59P2B5 transcript in spleen, kidney and small intestine. Figure 43 shows expression of 159P2B5 in bladder cancer patient specimens. RNA was extracted from bladder cancer cell lines (CL; UM-UC-3, J82, SCaBER), normal bladder (NB), and bladder cancer patient tumors (T) isolated from bladder cancer patients. Northern blots with 10 μg of total RNA were probed with the 159P2B5 SSH sequence. Size standards in kilobases are indicated on the side. Results show expression of 159P2B5 in patient bladder cancer tissues, and in the SCaBER bladder cancer cell line, but not in normal bladder, nor in the other cancer cell lines tested.

Figure 44 shows expression of 161P2B7A by RT-PCR. First sfrand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate metastasis to lymph node (LN), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 161P2B7A, was performed at 26 and 30 cycles of amplification. Results show strong expression of 161P2B7A in lung cancer pool and pancreas cancer pool. Expression' was also detected in prostate metastasis to LN, prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, ovary cancer pool, breast cancer pool, and cancer metastasis pool. Very low expression was observed in vital pool 2 but not in vital pool 1.

Figure 45 shows expression of 161P2B7A in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA/lane, were probed with the 161P2B7A SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show very low expression of 161P2B7A in testis but not in the other normal tissues tested.

Figure 46 shows expression of 161P2B7A in Multiple Normal Tissues. An mRNA dot blot containing 76 different samples from human tissues was analyzed using a 161P2B7A SSH probe. Expression was not detected in any ofthe 76 normal tissues tested. The positive genomic DNA confrol showed very strong signal confirming the validity ofthe experiment.

Figure 47 shows expression of 161P2B7A in kidney cancer patient specimens. RNA was extracted from normal kidney (Nk), kidney cancer patient tumors (T) and their normal adjacent tissues (N) isolated from kidney cancer patients. Northern blots with 10 μg of total RNA were probed with the 161P2B7A SSH fragment. Size standards in kilobases are indicated on the side. Results show expression of two 161P2B7A transcripts, approximately 1.2 and 7 kb, in kidney cancer specimens but not in normal kidney.

Figure 48 shows expression of 161P2B7A in lung cancer patient specimens. RNA was extracted from lung cancer cell lines (CL), normal lung, lung tumors (T), and their normal adjacent tissues (NAT) isolated from lung cancer patients. Northern blot with 10 mg of total RNA lane was probed with the 161P2B7A fragment. Size standards in kilobases (kb) are indicated on the side. The results show expression of 161P2B7A in the lung tumors, but not in normal lung tissues. Expression was also detected in the lung cancer cell lines CALU-1, A427 and NCI- 146 but not in the small cell lung cancer cell line NCI-H82.

Figure 49 shows expression of 161P2B7A in pancreas and ovary cancer patient specimens. RNA was extracted from normal pancreas (NPa), pancreas cancer (PC), normal ovary (NO), and ovary cancer patient specimen (OC). Northern blot with 10 mg of total RNA/lane was probed with the 161P2B7A fragment. Size standards in kilobases (kb) are indicated on the side. The results show expression of 161P2B7A in the pancreas and ovary cancer patient specimens, but not in the normal tissues.

Figure 50 shows expression of 179P3G7 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), bladder cancer pool, kidney cancer pool, lung cancer pool, breast cancer pool, cancer metastasis pool, pancreas cancer pool and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 179P3G7, was performed at 26 and 30 cycles of amplification. Results show strong expression of 179P3G7 in kidney cancer pool and breast cancer pool. Expression was also detected in bladder cancer pool, lung cancer pool, cancer metastasis pool, pancreas cancer pool and prostate metastasis to LN, and vital pool 1, but not in vital pool 2.

Figure 51 shows expression of 179P3G7 in normal tissues. Two multiple tissue northern blots (Clontech), botii with 2 μg of mRNA/lane, were probed with the 179P3G7 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of 179P3G7 sfrongly in skeletal muscle, and weakly in kidney, liver and heart but not in the other normal tissues tested.

Figure 52 shows expression of 179P3G7 in kidney cancer patient specimens. RNA was extracted from normal kidney (Nk), kidney cancer patient tumors (T) and their normal adjacent tissues (N) isolated from kidney cancer patients. Northern blots with 10 μg of total RNA were probed with the 179P3G7 SSH fragment. Size standards in kilobases are indicated on the side. Results show expression of 179P3G7 in kidney cancer specimens. Expression of 179P3G7 is stronger in kidney tumors compared to normal kidney tissues.

Figure 53 shows expression of 184P3C10B by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC prostate cancer xenograft pool (LAPC- 4AD, LAPC-4AI, LAPC-9 AD and LAPC-9 Al), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, and cancer metastasis pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 184P3C10B, was performed ^• at 26 and 30 cycles of amplification. Results show expression of 184P3C10B in xenograft pool, prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, and cancer metastasis pool. Expression was also detected in vital pool 2 but at a much lower level in vital pool 1.

Figure 54 shows expression of 184P3C10B in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA lane, were probed with the 184P3C10B SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of approximately 2.4 and 5 kb 184P3C10B transcripts in placenta and to lower level in colon and small intestine, but not in the other normal tissues tested.

Figure 55 shows expression of 184P3C10B in bladder cancer patient specimens. RNA was extracted from bladder cancer cell lines (CL; UM-UC-3, J82, SCaBER), normal bladder (Nb), bladder cancer patient tumors (T) and their normal adjacent tissue (N) isolated from bladder cancer patients. Northern blots with 10 μg of total RNA were probed with the 184P3C10B SSH sequence. Size standards in kilobases are indicated on the side. Results show expression of 184P3C10B in patient bladder cancer tissues, and in the bladder cancer cell line SCaBER, but not in normal bladder nor in the other bladder cancer cell lines tested.

Figure 56 shows expression of 184P3G10 by RT-PCR. First sfrand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), LAPC prostate cancer xenograft pool (LAPC- 4AD, LAPC-4AI, LAPC-9 AD and LAPC-9 Al), bladder cancer pool, kidney cancer pool, colon cancer pool, and lung cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 184P3G10, was performed at 26 and 30 cycles of amplification. Results show strong expression of 184P3G10 in bladder cancer pool, kidney cancer pool, and colon cancer pool. Expression was also detected in xenograft pool, lung cancer pool, vital pool 2 but not in vital pool 1.

Figure 57 shows expression of 184P3G10 in normal tissues. Two multiple tissue northern blots (Clontech) both with 2 μg of mRNA lane, were probed with the 184P3G10 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of two approximately 4.4kb 184P3G10 franscripts in colon and small intestine, but not in the other normal tissues tested. Figure 58 shows expression of 184P3G10 in patient cancer specimens and normal tissues. RNA was extracted from a pool of three bladder cancers, colon cancers, lung cancers, breast cancers, ovary cancers, cancer metastasis, as well as from normal prostate (NP), normal bladder (NB), and normal kidney (NK). Northern blot with 10 mg of total RNA/lane was probed with 184P3G10 sequence. Size standards in kilobases (kb) are indicated on the side. Results show strong expression of 184P3G10 in the bladder cancers, colon cancers and ovary cancers. Expression of 184P3G10 was also detected in lung cancers, breast cancers, and cancer metastasis but not in the normal tissues tested.

Figure 59 shows expression of 184P3G10 in bladder cancer patient specimens. RNA was extracted from bladder cancer cell lines (CL; UM-UC-3, J82, SCaBER), normal bladder (N), bladder cancer patient tumors (T) and their normal adjacent tissue (Nat) isolated from bladder cancer patients. Northern blots with 10 μg of total RNA were probed with the 184P3G10 SSH sequence. Size standards in kilobases are indicated on the side. Results show expression of 184P3G10 in patient bladder cancer tissues, but not in normal bladder nor in the bladder cancer cell lines tested.

Figure 60 shows expression of 185P2C9 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate metastasis to lymph node (LN), prostate cancer pool, bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 185P2C9, was performed at 30 cycles of amplification. Results show strong expression of 185P2C9 in bladder cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool and pancreas cancer pool. Expression was also detected in prostate metastasis to LN, prostate cancer pool, kidney cancer pool, breast cancer pool, cancer metastasis pool, vital pool 2 but not in vital pool 1.

Figure 61 shows expression of 185P2C9 in normal tissues. Two multiple tissue northern blots (Clontech), both with 2 μg of mRNA/lane, were probed with the 185P2C9 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of and approximately 8.5 kb 185P2C9 transcript in testis and brain, but not in the other normal tissues tested.

Figure 62 shows expression of 185P2C9 in bladder cancer patient specimens. RNA was extracted from bladder cancer cell lines (CL; UM-UC-3, J82, SCaBER), normal bladder (Nb), bladder cancer patient tumors (T) and their normal adjacent tissue (N) isolated from bladder cancer patients. Northern blots with 10 μg of total RNA were probed with the 185P2C9 SSH sequence. Size standards in kilobases are indicated on the side. Results show expression of 185P2C9 in bladder cancer patient tissues, and in the bladder cancer cell lines tested. Expression of 185P2C9 is significantly stronger in bladder tumor tissues compared to normal tissues.

Figure 63 shows expression of 185P2C9 in kidney cancer patient specimens. RNA was extracted from kidney cancer cell lines (CL; 769-P, A498, Caki-1), normal kidney (N), kidney cancer patient tumors (T) and their normal adjacent tissues (NAT) isolated from kidney cancer patients. Northern blots with 10 μg of total RNA were probed with the 185P2C9 SSH fragment. Size standards in kilobases are indicated on the side. Results show expression of 185P2C9 in kidney cancer specimens and kidney cancer cell lines, but not in normal kidney.

Figure 64 shows expression of 186P1H9 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), bladder cancer pool, kidney cancer pool, colon cancer pool, lung cancer pool, ovary cancer pool, cancer metastasis pool, and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 186P1H9, was performed at 26 and 30 cycles of amplification. Results show sfrong expression of 186P1H9 in kidney cancer pool, colon cancer pool, ovary cancer pool, cancer metastasis pool, and pancreas cancer pool. Expression was also detected in bladder cancer pool, lung cancer pool, and vital pool 2 but not in vital pool 1.

Figure 65 shows expression of 186P1H9 in normal tissues. Two multiple tissue northern blots (Clontech) both with 2 μg of mRNA lane, were probed with the 186P1H9 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of an approximately 2.6 kb 186P1H9 transcript in testis, spleen, pancreas and brain. Lower expression is also detected in heart, skeletal muscle, prostate, colon and small intestine.

Figure 66 shows expression of 186P1H9 in patient cancer specimens and normal tissues. RNA was extracted from a pool of three kidney cancers (KC), ovary cancers (OC), cancer metastasis (Met), pancreas cancers (PaC), as well as from normal prostate (NP), normal bladder (NB), and normal kidney (NK), normal colon (NC), normal lung (NL), normal breast (NBr), normal ovary (NO), and normal pancreas (NPa). Northern blot with 10 mg of total RNA lane was probed with 186P1H9 sequence. Size standards in kilobases (kb) are indicated on the side. Results show sfrong expression of 186P1H9 in the bladder cancers, ovary cancers, cancer metastasis and pancreas cancers, but not in normal tissues. Expression of 186P1H9 is significantly stronger in patient cancer tissues compared to normal tissues.

Figure 67 shows expression of 186P1H9 in kidney cancer patient specimens. RNA was extracted from kidney cancer cell lines (CL; 769-P, A498, Caki-1), normal kidney (N), kidney cancer patient tumors (T) and their normal adjacent tissues (NAT) isolated from kidney cancer patients. Northern blots with 10 μg of total RNA were probed with the 186P1H9 SSH fragment. Size standards in kilobases are indicated on the side. Results show strong expression of 186P 1 H9 in kidney cancer patient specimens, but not in normal kidney, nor in the kidney cancer cell lines.

Figure 68 shows expression of 186P1H9 in ovarian and testicular cancer patient specimens. RNA was extracted from normal ovary (NO), ovary cancer patient specimens (PI, P2, P3), normal testis (NTe), and testis cancer patient specimens (P4, P5, P6). Northern blot with 10 mg of total RNA/lane was probed with the 186P1H9 SSH fragment. Size standards in kilobases (kb) are indicated on the side. The results show strong expression of 186P1H9 in the ovary cancer patient specimens, but not in the normal ovary. Expression was also detected in normal and in testis cancer specimens.

Figure 69 shows expression of 187P3F2 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), kidney cancer pool, and pancreas cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers ^'■ to 187P3F2, was performed at 26 and 30 cycles of amplification. Results show strong expression of 187P3F2 in kidney cancer pool, pancreas cancer pool and vital pool 1, but not in vital pool 2.

Figure 70 shows expression of 187P3F2 in normal tissues. Two multiple tissue northern blots (Clontech) both with 2 μg of mRNA/lane, were probed with the 187P3F2 SSH fragment. Size standards in kilobases (kb) are indicated on the side. Results show expression of a 4.5 kb 187P3F2 transcript in kidney and brain, but not in the other tissues tested.

Figure 71 shows expression of 187P3F2 in patient cancer specimens and normal tissues. RNA was extracted from a pool of three kidney cancers (KC), pancreas cancers (PaC), as well as from normal prostate (NP), normal bladder (NB), and normal kidney (NK), normal colon (NC), normal lung (NL), normal breast (NBr), normal ovary (NO), and normal pancreas (NPa). Northern blot with 10 mg of total RNA lane was probed with 187P3F2 sequence. Size standards in kilobases (kb) are indicated on the side. Results show strong expression of 187P3F2 in kidney cancers, pancreas cancers, and normal kidney, but not in the other normal tissues. Figure 72 shows expression of 187P3F2 in pancreas cancer patient specimens. RNA was extracted from pancreas cancer cell lines (CL), normal pancreas (N), and pancreas tumor tissues (T) isolated from pancreatic cancer patients. Northern blot with 10 mg of total RNA/lane was probed with the 187P3F2 SSH fragment. Size standards in kilobases (kb) are indicated on the side. The results show strong expression of 187P3F2 in the pancreas cancer specimens, but not in normal pancreas nor in the cancer cell lines tested.

Figure 73 shows expression of 192P2G7 by RT-PCR. First sfrand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), prostate cancer pool, bladder cancer pool, kidney cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool, pancreas cancer pool, and prostate metastasis to lymph node (LN). Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 186P1H9, was performed at 26 and 30 cycles of amplification. Results show strong expression of 186P1H9 in pancreas cancer pool and prostate metastasis to LN. Expression was also detected in prostate cancer pool, bladder cancer pool, kidney cancer pool, lung cancer pool, ovary cancer pool, breast cancer pool, cancer metastasis pool, vital pool 2 but not in vital pool 1.

Figure 74 shows expression of 185P3C2 by RT-PCR. First strand cDNA was prepared from vital pool 1 (liver, lung and kidney), vital pool 2 (pancreas, colon and stomach), and bladder cancer pool. Normalization was performed by PCR using primers to actin and GAPDH. Semi-quantitative PCR, using primers to 185P3C2, was performed at 26 and 30 cycles of amplification. Results show strong expression of 185P3C2 in bladder cancer pool. Low level expression was detected in vital pool 2, but not in vital pool 1.

Example 5: Transcript Variants of Genes of the Invention

Transcript variants are variants of matured mRNA from the same gene by alternative transcription or alternative splicing. Alternative transcripts are transcripts from the same gene but start transcription at different points. Splice variants are mRNA variants spliced differently from the same transcript. In eukaryotes, when a multi-exon gene is transcribed from genomic DNA, the initial RNA is spliced to produce functional mRNA, which has only exons and is used for translation into an amino acid sequence. Accordingly, a given gene can have zero to many alternative transcripts and each franscript 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 the 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 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 and assembled into a consensus sequence. The original gene sequence is compared to the consensus sequence(s) or other full-length sequences. Each consensus sequence is a potential splice variant for that gene (see, e.g., the URL www.doubletwist.com/products/cl l_agentsOverview.jhtml). Even when a variant is identified that is not a full- length clone, that portion ofthe variant is very useful for antigen generation and for further cloning ofthe full- length splice variant, using techniques known in the art.

Moreover, computer programs available in the art are used that identify franscript 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 (at the World Wide Web URL: compbio.ornl.gov/Grail-bin/EmptyGrailForm) and GenScan (at the World Wide Web URL: genes.mit.edu/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 available in the art are used, such as full-length cloning, proteomic validation, PCR-based validation, and 5' RACE validation, etc. (see e.g., Proteomic Validation: Brennan, S.O., et al, Albumin banks peninsula: a new termination variant characterized by elecfrospray mass spectrometry, Biochem Biophys Acta. 1999 Aug 17;1433(l-2):321-6; Feπanti P, et al, Differential splicing of pre-messenger RNA produces multiple forms of mature caprine alρha(sl)-casein, Eur J Biochem. 1997 Oct l;249(l):l-7. For PCR-based Validation: Wellmann S, et al, Specific reverse transcription- PCR quantification of vascular endothelial growth factor (VEGF) splice variants by LightCycler technology, Clin Chem. 2001 Apr;47(4):654-60; Jia, H.P., et al, Discovery 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 ofthe murine reduced folate carrier gene and identification of variant splice forms, Biochem Biophys Acta. 1997 Aug 7; 1353(2): 191-8).

It is known in the art that genomic regions are modulated in cancers. When the genomic region to which a gene maps is modulated in a particular cancer, the alternative transcripts or splice variants ofthe gene are modulated as well. Disclosed herein is a particular expression profile ofthe target genes related to cancer. Alternative transcripts and splice variants of these genes may also be involved in cancers in the same or different tissues, thus serving as tumor-associated markers/antigens.

Using the full-length gene and EST sequences, five franscript variants were identified for 83P4B8 , seven for 109P1D4, one for 151P4E11, two for 161P2B7A, one for 179P3G7, four for 184P3G10, two for 185P2C9, four for 185P3C2, and two for 192P2G7, as displayed in Figures 11-14.

Figure 11 through Figure 14 are set forth herein on a gene-by-gene basis. The following list shows the numbering of figures and the coπesponding genes, nucleotide sequence of a franscript variant. Figure 11 displays the nucleotide sequences of franscript variants. Figure 12 shows amino acid sequences of proteins translated from the coπesponding transcript variants. Figure 13 displays the alignment of nucleotide sequences of franscript variants. Figure 14 displays the alignment of protein sequences from the coπesponding transcript variants.

Number of transcript variants for target genes and the numbering of associated igures .

Example 6: Production of Recombinant Targets ofthe Invention in Prokaryotic Systems

To express a recombinant gene of Figure 2 in prokaryotic cells, full or partial length gene cDNA sequences are cloned into any one of a variety of expression vectors known in the art. One or more ofthe following regions of genes set forth in Figure 2, or variants or analogs thereof, are expressed in these constructs: regions that encode the entire, respective, amino acid sequence of a particular target, or any 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 or more contiguous amino acids from a protein of Figure 2, variants, or analogs thereof.

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

B. Bacterial Constructs: pGEX Constructs: To generate recombinant proteins ofthe invention in bacteria that are fused to the Glutathione S-transferase (GST) protein, all of or parts of a cDNA protein coding sequence ofthe invention are fused to the GST gene by cloning into pGEX-6P-l or any other GST- fusion vector ofthe pGEX family (Amersham Pharmacia Biotech, Piscataway, NJ). These constracts allow controlled expression of recombinant target ofthe invention protein sequences with GST fused at the amino-terminus and a six histidine epitope (6X His) at the carboxyl-terminus. The GST and 6X His tags permit purification ofthe recombinant fusion protein from induced bacteria with the appropriate affinity matrix and allow recognition ofthe 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., ofthe open reading frame (ORF). A proteolytic cleavage site, such as the PreScission™ recognition site in pGEX-6P-l, can be employed to permit cleavage ofthe GST tag from target ofthe invention-related protein. The ampicillin resistance gene and pBR322 origin permits selection and maintenance ofthe pGEX plasmids in _5. coli. pMAL Constructs: To generate, in bacteria, recombinant target ofthe invention proteins that are fused to maltose-binding protein (MBP), all of or parts ofthe target ofthe invention cDNA protein coding sequence are fused to the MBP gene by cloning into the pMAL-c2X and ρMAL-p2X vectors (New England Biolabs, Beverly, MA). These constructs allow controlled expression of recombinant target ofthe invention protein sequences with MBP fused at the amino-terminus and a 6X His epitope tag at the carboxyl-terminus. The MBP and 6X His tags permit purification ofthe recombinant protein from induced bacteria with the appropriate affinity matrix and allow recognition ofthe 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 a target ofthe invention. The pMAL-c2X and ρMAL-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 a target ofthe invention in bacterial cells, all of or parts ofthe target ofthe invention cDNA protein coding sequence are cloned into the pET family of vectors (Novagen, Madison, WI). These vectors allow tightly controlled expression of recombinant target ofthe invention protein in bacteria with and without fusion to proteins that enhance solubility, such as NusA and tliioredoxin (Trx), and epitope tags, such as 6X His and S-Tag ™ that aid purification and detection ofthe recombinant protein. For example, constructs are made utilizing pET NusA fusion system 43.1 such that regions ofthe target ofthe invention protein are expressed as amino-terminal fusions to NusA. In one embodiment, a NusA-fusion protein encompassing certain amino acids of a Figure 2 protein with a C-terminal 6xHis tag are expressed in E. coli, purified by metal chelate affinity chromatography, and used as an immunogen for generation of antibodies.

C. Yeast Constructs: pESC Constructs: To express a target ofthe invention in the yeast species Saccharomyces cerevisiae for generation of recombinant protein and functional studies, all of or parts of a target of the invention cDNA protein coding sequence are cloned into the pESC family of vectors each of which contain 1 of 4 selectable markers, HIS3, TRP1, LEU2, and URA3 (Stratagene, La Jolla, CA). These vectors allow controlled expression from the same plasmid of up to 2 different genes or cloned sequences containing either Flag™ or Myc epitope tags in the same yeast cell. This system is useful to confirm protein-protein interactions of a target ofthe invention. In addition, expression in yeast yields similar post-translational modifications, such as glycosylations and phosphorylations, that are found when expressed in eukaryotic cells. pESP Constracts: To express a target ofthe invention in the yeast species Saccharomyces pombe, all of or parts of a target ofthe invention cDNA protein coding sequence are cloned into the pESP family of vectors. These vectors allow controlled high level of expression of a target ofthe invention protein sequence that is fused at either the amino teπninus or at the carboxyl terminus to GST which aids purification ofthe recombinant protein. A Flag™ epitope tag allows detection ofthe recombinant protein with anti- Flag™ antibody.

Example 7: Production of Recombinant Target of the Invention in Eukaryotic Systems A. Mammalian Constructs:

To express a recombinant target ofthe invention in eukaryotic cells, the full or partial length target ofthe invention cDNA sequences can be cloned into any one of a variety of expression vectors known in the art. One or more of the following peptide regions of a protein ofthe invention are expressed in these constructs: any 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 or more contiguous amino acids from a protein of Figure 2, variants, or analogs thereof. In certain embodiments a region of a specific variant of a target ofthe invention is expressed that encodes an amino acid at a specific position which differs from the amino acid of any other respective variant found at that position. In other embodiments, a region of a variant ofthe invention is expressed that lies partly or entirely within a sequence that is unique to that variant respective to other variants of that target.

The constructs can be transfected into any one of a wide variety of mammalian cells such as 293T cells. Transfected 293T cell lysates can be probed with the anti-target ofthe invention polyclonal serum, described herein. pcDNA4/HisMax Constructs: To express a target ofthe invention in mammalian cells, a target ofthe invention ORF, or portions thereof, are cloned into pcDNA4/HisMax Version A (Invitrogen, Carlsbad, CA). Protein expression is driven from the cytomegaloviras (CMV) promoter and the SP16 translational enhancer. The recombinant protein has Xpress™ and six histidine (6X His) epitopes fused to the amino-terminus. The pcDNA4/HisMax vector also contains the bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability along with the SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen. The Zeocin resistance gene allows for selection of mammalian cells expressing the protein and the ampicillin resistance gene and ColEl origin permits selection and maintenance ofthe plasmid in E. coli. pcDNA3.1/MvcHis Constructs: To express a target ofthe invention in mammalian cells, a target ofthe invention ORF, or portions thereof, with a consensus Kozak translation initiation site are cloned into ρcDNA3.1/MycHis Version A (Invitrogen, Carlsbad, CA). Protein expression is driven from the cytomegalovirus (CMV) promoter. The recombinant proteins have the myc epitope and 6X His epitope fused to the carboxyl- terminus. The pcDNA3.1/MycHis vector also contains the bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability, along with the SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen. The Neomycin resistance gene can be used, as it allows for selection of mammalian cells expressing the protein and the ampicillin resistance gene and ColEl origin permits selection and maintenance ofthe plasmid in E. coli.

PCDNA3.1/CT-GFP-TOPO Construct: To express a target ofthe invention in mammalian cells and to allow detection ofthe recombinant proteins using fluorescence, a target ofthe invention ORF, or portions thereof, with a consensus Kozak translation initiation site are cloned into pcDNA3.1/CT-GFP-TOPO (Invitrogen, CA). Protein expression is driven from the cytomegalovirus (CMV) promoter. The recombinant proteins have the Green Fluorescent Protein (GFP) fused to the carboxyl-terminus facilitating non-invasive, in vivo detection and cell biology studies. The pcDNA3.1CT-GFP-TOPO vector also contains the bovine growth hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability along with the SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen. The Neomycin resistance gene allows for selection of mammalian cells that express the protein, and the ampicillin resistance gene and ColEl origin permits selection and maintenance ofthe plasmid in E. coli. Additional constructs with an amino- terminal GFP fusion are made in pcDNA3.1/NT-GFP-TOPO spanning the entire length of a target ofthe invention protein.

PAPtag: A target ofthe invention ORF, or portions thereof, is cloned into pAPtag-5 (GenHunter Corp. Nashville, TN). This construct generates an alkaline phosphatase fusion at the carboxyl-terminus of a target ofthe invention protein while fusing the IgGκ signal sequence to the amino-terminus. Constructs are also generated in which alkaline phosphatase with an ammo-terminal IgG signal sequence is fused to the amino-terminus of a target ofthe invention protein. The resulting recombinant target ofthe invention proteins are optimized for secretion into the media of transfected mammalian cells and can be used to identify proteins such as ligands or receptors that interact with a target ofthe invention protein. Protein expression is driven from the CMV promoter and the recombinant proteins also contain myc and 6X His epitopes fused at the carboxyl-terminus that facilitates detection and purification. The Zeocin resistance gene present in the vector allows for selection of mammalian cells expressing the recombinant protein and the ampicillin resistance gene permits selection ofthe plasmid in E. coli. ptag5: A target ofthe invention ORF, or portions thereof, is cloned into pTag-5. This vector is similar to pAPtag but without the alkaline phosphatase fusion. This construct generates a target ofthe invention protein with an amino-terminal IgGκ signal sequence and myc and 6X His epitope tags at the carboxyl-terminus that facilitate detection and affinity purification. The resulting recombinant target ofthe invention protein is optimized for secretion into the media of transfected mammalian cells, and is used as immunogen or ligand to identify proteins such as ligands or receptors that interact with target ofthe invention proteins. Protein expression is driven from the CMV promoter. The Zeocin resistance gene present in the vector allows for selection of mammalian cells expressing the protein, and the ampicillin resistance gene permits selection ofthe plasmid in E. coli. PsecFc: A target ofthe invention ORF, or portions thereof, is also cloned into psecFc. The psecFc vector was assembled by cloning the human immunoglobulin GI (IgG) Fc (hinge, CH2, CH3 regions) into pSecTag2 (Invitrogen, California). This construct generates an IgGl Fc fusion at the carboxyl-terminus of a target ofthe invention protein, while fusing the IgGK signal sequence to N-terminus. Target ofthe invention fusions utilizing the murine IgGl Fc region are also used. The resulting recombinant target ofthe invention proteins are optimized for secretion into the media of transfected mammalian cells, and can be used as immunogens or to identify proteins such as ligands or receptors that interact with a target ofthe invention protein. Protein expression is driven from the CMV promoter. The hygromycin resistance gene present in the vector allows for selection of mammalian cells that express the recombinant protein, and the ampicillin resistance gene permits selection ofthe plasmid in E. coli. pSRoc Constructs: To generate mammalian cell lines that express a target ofthe invention constitutively, a target ofthe invention ORF, or portions thereof, are cloned into pSRα constracts. Amphotropic and ecotropic retroviruses are generated by transfection of pSRα constracts into the 293T-10A1 packaging line or co-transfection of pSRα 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 ofthe cloned gene, a target ofthe invention, into the host cell-lines. Protein expression is driven from a long terminal repeat (LTR). The Neomycin resistance gene present in the vector allows for selection of mammalian cells that express the protein, and the ampicillin resistance gene and ColEl origin permit selection and maintenance ofthe plasmid in E. coli. The retroviral vectors can thereafter be used for infection and generation of various cell lines using, for example, PC3, NIH 3T3, TsuPrl, 293 or rat-1 cells.

Additional pSRα constructs are made that fuse an epitope tag such as the FLAG™ tag to the carboxyl- terminus of a target ofthe invention sequence to allow detection using anti-Flag antibodies. For example, the

FLAG™ sequence 5' gat tac aag gat gac gac gat aag 3' (SEQ ID NO: ) is added to cloning primer at the 3' end ofthe ORF. Additional pSRα constructs are made to produce both arnino-terminal and carboxyl-terminal GFP and myc/6X His fusion proteins ofthe full-length target ofthe invention proteins.

Additional Viral Vectors: Additional constructs are made for viral-mediated delivery and expression of a target ofthe invention. High virus titer leading to high level expression of a target ofthe invention is achieved in viral delivery systems such as adenoviral vectors and heφes amplicon vectors. A target ofthe invention coding sequence or fragments thereof is 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, target ofthe invention 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 a target ofthe invention in mammalian cells, coding sequences of a target ofthe invention, or portions thereof, are cloned into regulated mammalian expression systems such as the T-Rex System (Invitrogen), the GeneSwitch System (Invitrogen) and the tightly-regulated Ecdysone System (Sratagene). These systems allow the study ofthe temporal and concentration dependent effects of recombinant targets ofthe invention. These vectors are thereafter used to confrol expression of a target ofthe invention in various cell lines such as PC3, NIH 3T3, 293 or rat-1 cells.

B. Baculovirus Expression Systems

To generate recombinant target ofthe invention proteins in a baculoviras expression system, a target ofthe invention ORF, or portions thereof, are cloned into the baculoviras transfer vector pBlueBac 4.5 (Invitrogen), which provides a His-tag at the N-terminus. Specifically, pBlueBac-target ofthe invention nucleic acid sequence is co- ttansfected with helper plasmid pBac-N-Blue (Invitrogen) into SF9 (Spodoptera frugiperda) insect cells to generate recombinant baculoviras (see Invitrogen instruction manual for details). Baculoviras is then collected from cell supernatant and purified by plaque assay.

Recombinant target ofthe invention protein is then generated by infection of HighFive insect cells (Invifrogen) with purified baculovirus. Recombinant target ofthe invention protein can be detected using anti- target of the invention or anti-His-tag antibody. Target of the invention protein can be purified and used in various cell-based assays or as immunogen to generate polyclonal and monoclonal antibodies specific for a target ofthe invention.

Example 8: Antigenicity Profiles and Secondary Structure

Figure 5, Figure 6, Figure 7, Figure 8, and Figure 9 depict graphically five amino acid profiles ofthe target ofthe invention amino acid sequences, each assessment available by accessing the ProtScale website (URL www.expasy.ch/cgi-bin/protscale.pl) 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 J., 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 Ponnuswamy P.K., 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 ofthe target ofthe invention proteins. Each ofthe above amino acid profiles were generated using the following ProtScale parameters for analysis: 1) A window size of 9; 2) 100% weight ofthe window edges compared to the window center; and, 3) amino acid profile values normalized to lie between 0 and 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 the surface ofthe protein, and thus be 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 for immune recognition, such as by antibodies.

Antigenic sequences ofthe target ofthe invention proteins indicated, e.g., by the profiles set forth in Figure 5, Figure 6, Figure 7, Figure 8, and/or Figure 9 are used to prepare immunogens, either peptides or nucleic acids that encode them, to generate therapeutic and diagnostic anti-target ofthe invention 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 coπesponding nucleic acids that encode them, from the target ofthe invention variant proteins. In particular, peptide immunogens for target ofthe invention proteins can comprise, a peptide region of at least 5 amino acids of a protein ofthe invention in any whole number increment up to an entire protein that includes an amino acid position having a value greater than 0.5 in the Hydrophilicity profile of Figure 5; a peptide region of at least 5 amino acids of a protein ofthe invention in any whole number increment up to the entire protein that includes an amino acid position having a value less than 0.5 in the Hydropathicity profile of Figure 6; a peptide region of at least 5 amino acids of a protein ofthe invention in any whole number increment up to the entire protein that includes an amino acid position having a value greater than 0.5 in the Percent Accessible Residues profile of Figure 7; a peptide region of at least 5 amino acids of a protein ofthe invention in any whole number increment up to the entire protein that includes an amino acid position having a value greater than 0.5 in the Average Flexibility profile on Figure 8; and, a peptide region of at least 5 amino acids of a protein ofthe invention in any whole number increment up to the entire protein that includes an amino acid position having a value greater than 0.5 in the Beta-rum profile of Figure 9.

All immunogens ofthe invention, whether peptides or nucleic acids, can be embodied in human unit dose form, or comprised by a composition that includes a pharmaceutical excipient compatible with human physiology.

The secondary structure of a protein ofthe invention, namely the predicted presence and location of alpha helices, extended strands, and random coils, is predicted from the primary amino acid sequence using the HNN - Hierarchical Neural Network method (Guermeur, 1997, World Wide Web URL pbil.ibcp.fr/cgi- bin/npsa_automat.pl?page=npsa_nn.html), accessed from the ExPasy molecular biology server (World Wide Web URL www.expasy.ch/tools/). The analysis provides the data set forth in iFigure 10 on protein by protein basis.

Analysis for the presence of transmembrane domains in a protein ofthe invention was carried out using a variety of transmembrane prediction algorithms many of which were accessed from the ExPasy molecular biology server (World Wide Web URL www.expasy.ch/tools/). The programs provide the data summarized in Table XXI on a protein by protein basis.

Example 9: Generation of Polyclonal Antibodies of the Invention

Polyclonal antibodies can be raised in a mammal, for example, by one or more injections of an immunizing agent (e.g., a protein ofthe invention) 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 a full length protein ofthe invention such as that set forth in figure 2, computer algorithms are employed in design of immunogens that, based on amino acid sequence analysis contain characteristics of being antigenic and available for recognition by the immune system ofthe immunized host (see the Example entitled "Antigenicity Profiles"). Such regions would be predicted to be hydrophilic, flexible, in beta-rum conformations, and/or be exposed on the surface ofthe protein (see, e.g., Figure 5, Figure 6, Figure 7, Figure 8, and Figure 9 for amino acid profiles that indicate such regions of a protein ofthe invention).

For example, of Figure 2 recombinant bacterial fusion proteins or peptides containing hydrophilic, flexible, beta-turn regions, generally found in regions between transmembrane domains and at the amino and carboxyl termini, are used as antigens to generate polyclonal antibodies in New Zealand White rabbits. Examples of such regions can be extracellular or intracellular. In addition, the amino-terminal region of a variant that is not present in a respective variant can be used as an immunogen. Antibodies to such regions are useful to distinguish one variant protein from another variant of that target. It is useful to conjugate the immunizing agent to a protein known to be ^• immunogenic in the mammal being immunized. Examples of such immunogenic proteins include, but are not limited to, keyhole limpet hemocyanin (KLH), serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. In one embodiment, a peptide encoding amino acids from a protein ofthe invention is conjugated to KLH and used to immunize the rabbit. Alternatively the immunizing agent can include all or portions ofthe of a protein ofthe invention e.g. in Figure 2, analogs or fusion proteins thereof. For example, a Figure 2 amino acid sequence can 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 of a protein ofthe invention is produced and purified and used as immunogen. Other recombinant bacterial fusion proteins that can be employed include maltose binding protein, LacZ, thioredoxin, NusA, or an immunoglobulin constant region (see the Example entitled "Production of Recombinant Targets ofthe Invention in Prokaryotic Systems" and Cuπent Protocols In Molecular Biology, Volume 2, Unit 16, Frederick M. Ausubul et al. eds., 1995; Linsley, P.S., Brady, W., Umes, 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 Example entitled "Production of Recombinant Targets ofthe Invention in Eukaryotic Systems"), and retain post- translational modifications such as glycosylations found in native protein. In one embodiment, amino acids from a protein ofthe invention are 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-produced protein ofthe invention 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 ofthe host animal. Examples of adjuvants include, but are not limited to, complete Freund's adjuvant (CFA) and MPL-TDM adjuvant (monophosphoryl Lipid A, synthetic frehalose dicorynomycolate).

In a typical protocol, rabbits are initially immunized subcutaneously with up to 200 μg, typically 100-200 μg, 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 μg, typically 100-200 μg, ofthe 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 ofthe antiserum by ELISA. >

To test reactivity and specificity of immune serum, such as the rabbit serum derived from immunization with Tag5-produced protein ofthe invention, a full-length Figure 2 cDNA is cloned into pCDNA 3.1 myc-his expression vector (Invitrogen, see the Example entitled "Production of Recombinant Targets ofthe Invention in Eukaryotic Systems"). After transfection ofthe constructs into 293T cells, cell lysates are probed with the respective anti-protein ofthe invention antibodies and with anti-His antibody (Santa Cruz Biotechnologies, Santa Cruz, CA) to determine specific reactivity ofthe antibodies to the respective denatured protein ofthe invention using the Western blot technique. Immunoprecipitation and flow cytometric analyses of 293T and other recombinant of Figure 2-expressing cells determine recognition of native protein by the antibodies. In addition, Westem.blot, immunoprecipitation, fluorescent microscopy, and flow cytometric techniques using cells that endogenously express the protein ofthe invention are carried out to test specificity.

Anti-serum from rabbits immunized with target of the invention 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 iπelevant fusion protein. For example, antiserum derived from a GST-of a Figure 2 fusion protein 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 comprising those amino acids covalently coupled to Affigel matrix. The serum is then further purified by protein G affinity chromatography to isolate the IgG fraction. Sera from other His-tagged antigens and peptide immunized rabbits as well as fusion partner depleted sera are affinity purified by passage over a column matrix composed ofthe original protein immunogen or free peptide.

Example 10: Generation of Monoclonal Antibodies .mAbs) of the Invention

In one embodiment, therapeutic mAbs to a protein ofthe invention comprise those that react with epitopes ofthe protein that would disrupt or modulate the biological function of a protein ofthe invention, for example antibodies that disrupt its interaction with ligands and bmding partners. Therapeutic mAbs also comprise those that specifically bind epitopes of a protein ofthe invention exposed on the cell surface and thus are useful in targeting mAb-toxin conjugates. Immunogens for generation of such mAbs include those designed to encode or contain an entire protein ofthe invention, regions of a protein ofthe invention predicted to be antigenic from computer analysis ofthe amino acid sequence (see, e.g., Figure 5, Figure 6, Figure 7, Figure 8, or Figure 9, and the Example entitled "Antigenicity Profiles"), and regions such as extracellular domains. Immunogens include peptides, recombinant bacterial proteins, and mammalian expressed Tag 5 proteins and human and murine IgG FC fusion proteins. In addition, cells expressing high levels of a protein ofthe invention, such as 293T-protein ofthe invention or 300.19-protein ofthe invention murine Pre-B cells, are used to immunize mice.

To generate mAbs to a protein ofthe invention, mice are first immunized intraperitoneally (IP) with, typically, 10-50 μg of protein immunogen or 10⁷ protein ofthe invention-expressing cells mixed in complete Freund's adjuvant. Mice are then subsequently immunized IP every 2-4 weeks with, typically, 10-50 μg of protein ' immunogen or 10⁷ cells mixed in incomplete Freund's adjuvant. Alternatively, MPL-TDM adjuvant is used in immunizations. In addition to the above protein and cell-based immunization strategies, a DNA-based immunization protocol is employed in which a mammalian expression vector encoding a protein ofthe invention sequence is used to immunize mice by direct injection ofthe plasmid DNA. For example, amino acids from a protein ofthe invention are cloned into the Tag5 mammalian secretion vector and the recombinant vector is used as immunogen. In another example the same amino acids are cloned into an Fc-fusion secretion vector in which the protein, ofthe invention sequence is fused at the amino-terminus to an IgK leader sequence and at the carboxyl- terminus to the coding sequence ofthe human or murine IgG Fc region. This recombinant vector is then used as immunogen. The plasmid immunization protocols are used in combination with purified proteins expressed from the same vector and with cells expressing protein ofthe invention.

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

In one embodiment for generating monoclonal antibodies reactive with a protein ofthe invention, a Tag5- protein ofthe invention antigen is expressed and purified from stably transfected 293T cells. Balb C mice are initially immunized intraperitoneally with 25 μg ofthe Tag5-protein ofthe invention mixed in complete Freund's adjuvant. Mice are subsequently immunized every two weeks with 25 μg ofthe 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 protein ofthe invention is monitored by Western blotting, immunoprecipitation and flow cytometry using 293T cells transfected with an expression vector encoding the protein ofthe invention cDNA (see e.g., the Example entitled "Production of Recombinant Targets of the Invention in Eukaryotic Systems"). Other recombinant protein ofthe invention-expressing cells or cells endogenously expressing a protein ofthe invention 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 ofthe sacrificed mice are harvested and 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, immunoprecipitation, fluorescent microscopy, and flow cytometry to identify protein ofthe invention-specific antibody-producing clones.

The binding affinity of a monoclonal antibody is determined using standard technologies. Affinity measurements quantify the strength of antibody to epitope binding and are used to help define which monoclonal antibodies reactive with proteins ofthe invention are suitable for diagnostic or therapeutic use, as appreciated by one of skill in the art. The BIAcore system (Uppsala, Sweden) is a useful 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, Methods in Enzymology 295: 268) to monitor biomolecular interactions in real time. BIAcore analysis conveniently generates association rate constants, dissociation rate constants, equilibrium dissociation constants, and affinity constants.

Example 11: HLA Class I and Class II Binding Assays

HLA class I and class II binding assays using purified HLA molecules are performed in accordance with disclosed protocols (e.g., PCT publications WO 94/20127 and WO 94/03205; Sidney et al, Current Protocols in Immunology 18.3.1 (1998); Sidney, et al, J. Immunol 154:247 (1995); Sette, et al, Mol Immunol. 31:813 (1994)). Briefly, purified MHC molecules (5 to 500 nM) are incubated with various unlabeled peptide inhibitors and 1-10 nM ¹²⁵I-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 the concenfration of HLA molecules necessary to bind 10-20% ofthe total radioactivity. All subsequent inhibition and direct binding assays are performed using these HLA concentrations.

Since under these conditions [label]<[HLA] and ICso≥tHLA], the measured IC₅₀ values are reasonable approximations ofthe true K_D values. Peptide inhibitors are typically tested at concentrations ranging from 120 μg/ml to 1.2 ng/ml, and are tested in two to four completely independent experiments. To allow comparison ofthe data obtained in different experiments, a relative binding figure is calculated for each peptide by dividing the IC₅₀ of a positive control for inhibition by the IC₅₀ for each tested peptide (typically unlabeled versions ofthe 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 ofthe positive confrols for inhibition by the relative binding ofthe peptide of interest. This method of data compilation is accurate and consistent for comparing peptides that have been tested on different days, or with different lots of purified MHC.

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

Example 12: Identification of HLA Supermotif- and Motif-Bearing CTL Candidate Epitopes HLA vaccine compositions ofthe invention can include multiple epitopes. The multiple epitopes can comprise multiple HLA supermotifs or motifs to achieve broad population coverage. This example illustrates the identification and confirmation of supermotif- and motif-bearing epitopes for the inclusion in such a vaccine composition. Calculation of population coverage is performed using the strategy described below.

Computer searches and algorithms for identification of supermotif and/or motif-bearing epitopes

The searches performed to identify the motif-bearing peptide sequences in the Example entitled "Antigenicity Profiles" (and, e.g., Tables V-XVIII, and Tables XXIII to XXVI) employ the protein sequence data from the protein set forth in Figures 2 and 3.

Computer searches for epitopes bearing HLA Class I or Class II supermotifs or motifs are performed as follows. All translated Figure 2 protein sequences are analyzed using a text string search software program to identify potential peptide sequences containing appropriate HLA binding motifs; such programs are readily produced in accordance with information in the art in view of known motif/supermotif disclosures. Furthermore, such calculations can be made mentally.

Identified A2-, A3-, and DR-supermotif sequences are scored using polynomial algorithms to predict their capacity to bind to specific HLA-Class I or Class π molecules. These polynomial algorithms account for the impact of different amino acids at different positions, and are essentially based on the premise that the overall affinity (or ΔG) of peptide-HLA molecule interactions can be approximated as a linear polynomial function ofthe type:

"ΔG" = a« x a_2l- x a_3l- x a_m- where a,-,- is a coefficient which represents the effect of the presence of a given amino acid (/) at a given position (i ) 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 i in the peptide, it is assumed to contribute a constant amount./ to the free energy of binding ofthe peptide irrespective ofthe sequence ofthe rest ofthe peptide.

The method of derivation of specific algorithm coefficients has been described in Gulukota et al, J. Mol. Biol. 267:1258-126, 1997; (see also Sidney et al, Human Immunol. 45:79-93, 1996; and Southwood et al, J. Immunol. 160:3363-3373, 1998). Briefly, for all i positions, anchor and non-anchor alike, the geometric mean of the average relative binding (ARB) of all peptides carrying j is calculated relative to the remainder ofthe group, and used as the estimate of/,-. For Class II peptides, if multiple alignments^fare 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 coπesponding to the sequence ofthe peptide are multiplied. If this product exceeds a chosen threshold, the peptide is predicted to bind. Appropriate thresholds are chosen as a function ofthe degree of stringency of prediction desired.

Selection of HLA-A2 supertype cross-reactive peptides

Protein sequences from Figure 2 proteins are scanned utilizing motif identification software, to identify 8-, 9- 10- and 11-mer sequences containing the HLA-A2-supermotif main anchor specificity. Typically, these sequences are then scored using the protocol described above and the peptides coπesponding 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 that bind to at least three ofthe five A2-supertype alleles tested are typically deemed A2-supertype cross-reactive binders. Prefeπed 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-bearing epitopes

The Figure 2 protein sequence(s) scanned above are also examined for the presence of peptides with the HLA- A3 -supermotif primary anchors. Peptides coπesponding to the HLA A3 supermotif-bearing sequences are then synthesized and tested for binding to HLA-A*0301 and HLA- A* 1101 molecules, the molecules encoded by the two most prevalent A3-supertype alleles. The peptides that bind at least one ofthe 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 ofthe five HLA- A3-supertype molecules tested.

Selection of HLA-B7 supermotif bearing epitopes

The Figure 2 ρrotein(s) scanned above are also analyzed for the presence of 8-, 9- 10-, or 11-mer peptides with the HLA-B7-supermotif. Coπesponding peptides are synthesized and tested for binding to HLA-B*0702, the 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*3501, B*5101, B*5301, and B*5401). Peptides capable of binding to three or more ofthe five B7-suρertype 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 ofthe Figure 2 proteins is 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 13: Confirmation of Immunogenicity

Cross-reactive candidate CTL A2-supermotif-bearing peptides that are identified as described herein are selected to confirm in vitro immunogenicity. Confirmation is performed using the following methodology:

Target Cell Lines for Cellular Screening:

The .221A2.1 cell line, produced by transferring the HLA-A2.1 gene into the HLA-A, -B, -C null mutant human B-lymphoblastoid cell line 721.221, is used as the 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 pyravate, 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 the 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 μg/ml DNAse, washed twice and resuspended in complete medium (RPMI-1640 plus 5% AB human serum, non-essential a ino 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, the non-adherent cells are removed by gently shaking the plates and aspirating the supernatants. The wells are washed a total of three times with 3 ml RPMI to remove most ofthe non- adherent and loosely adherent cells. Three ml of complete medium containing 50 ng/ml of GM-CSF and 1,000 U/ml of IL-4 are then added to each well. TNFα is added to the DCs on day 6 at 75 ng/ml and the cells are used for CTL induction cultures on day 7.

Induction of CTL with DC and Peptide: CD8+ T-cells are isolated by positive selection with Dynal immunomagnetic beads (Dynabeads® M-450) and the detacha-bead® reagent. Typically about 200-250xl0⁶ PBMC are processed to obtain 24xl0⁶ CD8⁺ T-cells (enough for a 48-well plate culture). Briefly, the PBMCs are thawed in RPMI with 30μg/ml DNAse, washed once with 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 with PBS/AB serum, added to the cells (140μl beads/20xl0⁶ cells) and incubated for 1 hour at 4°C with continuous mixing. The beads and cells are washed 4x with PBS/AB serum to remove the nonadherent cells and resuspended at lOOxlO⁶ cells/ml (based on the original cell number) in PBS/AB serum containing lOOμl/ml detacha-bead® reagent and 30 μg/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 40μg/ml of peptide at a cell concentration of l-2xl0⁶/ml in the presence of 3μg/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 lxlO⁵ 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 the presence of 10 ng/ml of IL-7. Recombinant human IL-10 is added the next day at a final concentration of 10 ng/ml and rhuman IL-2 is added 48 hours later at 10 IU/ml.

Restimulation ofthe induction cultures with peptide-pulsed adherent cells: Seven and fourteen days after the primary induction, the cells are restimulated with peptide-pulsed adherent cells. The PBMCs are thawed and washed twice with RPMI and DNAse. The cells are resuspended at 5x10⁶ cells/ml and iπadiated at ~4200 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 RPMI by tapping the plate gently to remove the nonadherent cells and the adherent cells pulsed with lOμg/ml of peptide in the 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 ofthe media is aspirated from the induction cultures (CD8+ cells) and brought to 0.5 ml with fresh media. The cells are then transfeπed to the wells containing the peptide-pulsed adherent cells. Twenty four hours later recombinant human IL-10 is added at a final concentration of 10 ng/ml and recombinant human IL2 is added the next day and again 2-3 days later at 50IU/ml (Tsai et al, Critical Reviews in Immunology 18(l-2):65-75, 1998). Seven days later, the cultures are assayed for CTL activity in a ⁵¹Cr release assay. In some experiments the cultures are assayed for peptide-specific recognition in the in situ IFNγ ELISA at the time ofthe 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 lvtic activity bv ⁵¹Cr release.

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

Adherent target cells are removed from culture flasks with trypsin-EDTA. Target cells are labeled with 200μCi of ⁵¹Cr sodium chromate (Dupont, Wilmington, DE) for 1 hour at 37°C. Labeled target cells are' resuspended at 10* per ml and diluted 1:10 with K562 cells at a concentration of 3.3xl0⁶/ml (an NK-sensitive erythroblastoma cell line used to reduce non-specific lysis). Target cells (100 μl) and effectors (lOOμl) are plated in 96 well round-bottom plates and incubated for 5 hours at 37°C. At that time, 100 μl of supernatant are collected from each well and percent lysis is determined according to the formula:

[(cpm ofthe test sample- cpm ofthe spontaneous ⁵¹Cr release sample)/(cpm ofthe maximal ⁵¹Cr release sample- cpm ofthe spontaneous ⁵¹Cr release sample)] x 100.

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

In situ Measurement of Human IFNγ Production as an Indicator of Peptide-specific and Endogenous

Recognition

Immulon 2 plates are coated with mouse anti-human IFNy_ monoclonal antibody (4 μg/ml 0.1M NaHC0₃, pH8.2) overnight at 4°C. The plates are washed with Ca²⁺, Mg²⁺-free PBS/0.05% Tween 20 and blocked with PBS/10% FCS for two hours, after which the CTLs (100 μl/well) and targets (100 μl/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 concenfration 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 μl 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-stteptavidin ( 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 IM 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, 5x10⁴ CD8+ cells are added to a T25 flask containing the following: lxlO⁶ iπadiated (4,200 rad) PBMC (autologous or allogeneic) per ml, 2xl0⁵ iπadiated (8,000 rad) EBV- transformed cells per ml, and OKT3 (anti-CD3) at 30ng per ml in RPMI-1640 containing 10% (v/v) human AB serum, non-essential amino acids, sodium pyruvate, 25 μM 2-mercaptoethanol, L-glutamine and penicillin/streptomycin. Recombinant human IL2 is added 24 hours later at a final concenfration of 200IU/ml and every 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 ⁵¹Cr release assay or at lxl0⁶/ml in the in situ IFNγ 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 5x10⁴ 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 iπadiated (4,200 rad); 2x10⁵ iπadiated (8,000 rad) EBV-fransformed cells per ml RPMI- 1640 containing 10%(v/v) human AB serum, non-essential AA, sodium pyravate, 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 the ability to induce peptide-specific CTL in normal individuals. In this analysis, a peptide is typically considered to be an epitope if it induces peptide-specific CTLs in at least individuals, and preferably, also recognizes the endogenously expressed peptide.

Immunogenicity can also be confirmed using PBMCs isolated from patients bearing a tumor that expresses a Figure 2 protein. Briefly, PBMCs are isolated from patients, re-stimulated with peptide-pulsed monocytes and assayed for the ability to recognize peptide-pulsed target cells as well as transfected cells endogenously expressing the antigen.

Evaluation of A*03/A11 immunogenicity

HLA-A3 supermotif-bearing cross-reactive binding peptides are also evaluated for immunogenicity using methodology analogous for that used to evaluate the immunogenicity ofthe HLA-A2 supermotif peptides.

Evaluation of B7 immunogenicity

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

Peptides bearing other supermotifs/motifs, e.g., HLA-A1, HLA-A24 etc. are also confirmed using similar methodology

Example 14: 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 the identification and preparation of highly cross-reactive native peptides, as demonstrated herein. Moreover, the definition of HLA motifs and supermotifs also allows one to engineer highly cross-reactive epitopes by identifying residues within a native peptide sequence which can be analoged to confer upon the peptide certain characteristics, e.g. greater cross-reactivity within the group of HLA molecules that comprise a supertype, and/or greater binding affinity for some or all of those HLA molecules. Examples of analoging peptides to exhibit modulated binding affinity are set forth in this example.

Analoging at Primary Anchor Residues

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

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

Alternatively, a peptide is confirmed as binding one or all supertype members and then analoged to modulate binding affinity to any one (or more) ofthe supertype members to add population coverage.

The selection of analogs for immunogenicity in a cellular screening analysis is typically further restricted by the capacity ofthe parent wild type (WT) peptide to bind at least weakly, i.e., bind at an IC₅₀ of 5000nM or less, to three of more A2 supertype alleles. The rationale for this requirement is that the WT peptides must be present endogenously in sufficient quantity to be biologically relevant. Analoged peptides have been shown to have increased immunogenicity 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, Proc. 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 endogenously express the epitope.

Analoging of HLA- A3 and B7-supermotif-bearing peptides

Analogs of HLA- A3 supermotif-bearing epitopes are generated using strategies similar to those employed in analoging HLA-A2 supermotif-bearing peptides. For example, peptides binding to 3/5 ofthe A3-supertype molecules are engineered at primary anchor residues to possess a prefeπed residue (V, S, M, or A) at position 2.

The analog peptides are then tested for the ability to bind A*03 and A*l 1 (prototype A3 supertype alleles). Those peptides that demonstrate < 500 nM binding capacity are then confirmed as having A3-supertype cross- reactivity.

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

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

The analog peptides are then be confirmed for immunogenicity, typically in a cellular screening assay. Again, it is generally important to demonsfrate 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 highly cross-reactive peptides and/or peptides that bind HLA molecules with increased affinity by identifying particular residues at secondary anchor positions that are associated with such properties. For example, the binding capacity of a B7 supermotif-bearing peptide with an F residue at position 1 is analyzed. The peptide is then analoged to, for example, substitute L for F at position 1. The analoged peptide is evaluated for increased binding affinity, binding half life and/or increased cross-reactivity. Such a procedure identifies analoged peptides with enhanced properties.

Engineered analogs with sufficiently improved binding capacity or cross-reactivity can also be tested for immunogenicity in HLA-B7-transgenic mice, following for example, IFA immunization or lipopeptide iirimunization. Analoged peptides are additionally tested for the ability to stimulate a recall response using PBMC from patients with protein(s) of Figure 2-expressing tumors.

Other analoging strategies

Another form of peptide analoging, unrelated to anchor positions, involves the substitution of a cysteine with α-amino butyric acid. Due to its chemical nature, cysteine has the propensity to form disulfide bridges and sufficiently alter the peptide structurally so as to reduce binding capacity. Substitution of α-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 al, In: Persistent Viral Infections, Eds. R. Ahmed and I. Chen, John Wiley & Sons, England, 1999).

Thus, by the use of single amino acid substitutions, the binding properties and/or cross-reactivity of peptide ligands for HLA supertype molecules can be modulated. Example 15: Identification of HLA-DR binding motifs in Proteins of Figure 2 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-bearing epitopes.

To identify HLA class II HTL epitopes derived from a protein of Figure 2, a Figure 2 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- and C-terminal flanking regions (15 amino acids total).

Protocols for predicting peptide binding to DR molecules have been developed (Southwood 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 the presence of DR-supermotif primary anchors (i.e., at position 1 and position 6) within a 9-mer core, but also additionally evaluates sequences for the presence of secondary anchors. Using allele-specific selection tables (see, e.g., Southwood et al, ibid.), it has 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 DRl, DR4w4, and DR7, can efficiently select DR cross-reactive peptides.

The protein in Figure 2-derived peptides identified above are tested for their binding capacity for various common HLA-DR molecules. All peptides are initially tested for binding to the DR molecules in the primary panel: DRl, DR4w4, and DR7. Peptides binding at least two of these three DR molecules are then tested for binding to DR2w2 βl, DR2w2 β2, DR6wl9, and DR9 molecules in secondary assays. Finally, peptides binding at least two ofthe four secondary panel DR molecules, and thus cumulatively at least four of seven different DR molecules, are screened for binding to DR4wl5, DR5wl 1, and DR8w2 molecules in tertiary assays. Peptides binding at least seven ofthe ten DR molecules comprising the primary, secondary, and tertiary screening assays are considered cross-reactive DR binders. Proteins in Figure 2-derived peptides found to bind common HLA-DR alleles are of particular interest.

Selection of DR3 motif peptides

Because HLA-DR3 is an allele that is prevalent in Caucasian, Black, and Hispanic populations, DR3 binding capacity is a relevant criterion in the selection of HTL epitopes. Thus, peptides shown to be candidates may also be assayed for their DR3 binding capacity. However, in view ofthe binding specificity ofthe 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, Figure 2 antigens are analyzed for sequences carrying one ofthe two DR3-specific binding motifs reported by Geluk et al. (J. Immunol. 152:5742-5748, 1994). The coπesponding peptides are then synthesized and confirmed as having the ability to bind DR3 with an affinity of lμM. or better, i.e., less than 1 μM. Peptides are found that meet this binding criterion and qualify as HLA class II high affinity binders.

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

Similarly to the case of HLA class I motif-bearing peptides, the class II motif-bearing peptides are analoged to improve affinity or cross-reactivity. For example, aspartic acid at position 4 ofthe 9-mer core sequence is an optimal residue for DR3 binding, and substitution for that residue often improves DR 3 binding. Example 16: Immunogenicity of HTL epitopes derived from a Protein of Figure 2

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 proteins of Figure 2-expressing tumors.

Example 17: Calculation of phenotypic frequencies of HLA-supertypes in various ethnic backgrounds to determine breadth of population coverage

This example illustrates the assessment ofthe breadth of population coverage of a vaccine composition comprised of multiple epitopes comprising multiple supermotifs and/or motifs.

In order to analyze population coverage, gene frequencies of HLA alleles are deteπnined. 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, 1996). To obtain overall phenotypic frequencies, cumulative gene frequencies are calculated, and the cumulative antigen frequencies derived by the use ofthe inverse formula [af=l-(l-Cgf)²].

Where frequency data is not available at the level of DNA typing, coπespondence to the serologically defined antigen frequencies is assumed. To obtain total potential supertype population coverage no linkage disequilibrium is assumed, and only alleles confirmed to belong to each ofthe 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 ofthe non-A covered population that could be expected to be covered by the B alleles considered (e.g., total=A+B*(l-A)). Confirmed members ofthe A3-like supertype are A3, Al 1, A31, A*3301, and A*6801. Although the A3-like supertype may also include A34, A66, and A*7401, these alleles were not included in overall frequency calculations. Likewise, confirmed members ofthe A2-like supertype family are A*0201, A*0202, A*0203, A*0204, A*0205, A*0206, A*0207, A*6802, and A*6901. Finally, the B7-like supertype- confirmed 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-supertypes is approximately 86% in five major ethnic groups. Coverage may be extended by including peptides bearing the Al and A24 motifs. On average, Al is present in 12% and A24 in 29% ofthe population across five different major ethnic groups (Caucasian, North American Black, Chinese, Japanese, and Hispanic). Together, these alleles are represented with an average frequency of 39% in these same ethnic populations. The total coverage across the major ethnicities when Al and A24 are combined with the coverage ofthe 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 et al, J. Clin. Invest. 100:503, 1997; Doolan et al, Immunity 7:97, 1997; and Threlkeld et al, J. Immunol. 159:1648, 1997) have shown that highly cross-reactive binding peptides are almost always recognized as epitopes. The use of highly cross-reactive binding peptides is an important selection criterion in identifying candidate epitopes for inclusion in a vaccine that is immunogenic in a diverse population. With a sufficient number of epitopes (as disclosed herein and from the art), an average population coverage is predicted to be greater than 95% in each of five major ethnic populations. The game theory Monte Carlo simulation analysis, wliich is known in the art (see e.g., Osbome, MJ. and Rubinstein, A. "A course in game theory" MIT Press, 1994), can be used to estimate what percentage ofthe individuals in a population comprised of the Caucasian, North American Black, Japanese, Chinese, and Hispanic ethnic groups would recognize the vaccine epitopes described herein. A prefeπed percentage is 90%. A more prefeπed percentage is 95%.

Example 18: CTL Recognition Of Endogenously Processed Antigens After 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 fransgenic 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 further re-stimulated. An additional six days later, these cell lines are tested for cytotoxic activity on ⁵¹Cr labeled Jurkat-A2.1/K^b target cells in the absence or presence of peptide, and also tested on ⁵¹Cr labeled target cells bearing the endogenously synthesized antigen, i.e. cells that are stably transfected with a gene of Figure 2-related expression vector.

The results demonsfrate that CTL lines obtained from animals primed with peptide epitopes recognize endogenously synthesized Figure 2 antigens. The choice of transgenic mouse model to be used for such an analysis depends upon the eρitope(s) that are being evaluated. In addition to HLA-A*0201/K^b transgenic mice, several other transgenic mouse models including mice with human Al l, which may also be used to evaluate A3 epitopes, and B7 alleles have been characterized and others (e.g., fransgenic mice for HLA-A1 and A24) are being developed. HLA- DR1 and HLA-DR3 mouse models have also been developed, which may be used to evaluate HTL epitopes.

Example 19: Activity Of CTL-HTL Conjugated Epitopes In Transgenic Mice

This example illustrates the induction of CTLs and HTLs in transgenic mice, by use of a protein of Figure 2-derived CTL and HTL peptide vaccine compositions. The vaccine compositions used herein comprise peptides to be administered to a patient with a protein of Figure 2-expressing tumor. The peptide composition can comprise multiple CTL and/or HTL epitopes. The epitopes are identified using methodology as described herein. This example also illustrates that enhanced immunogenicity can be achieved by inclusion of one or more HTL epitopes in a CTL vaccine composition; such a peptide composition can comprise an HTL epitope conjugated to a CTL epitope. The CTL epitope can be one that binds to multiple HLA family members at an affinity of 500 nM or less, or analogs of that epitope. The peptides may be lipidated, if desired.

Immunization procedures: Immunization of transgenic mice is performed as described (Alexander et al, J. Immunol. 159:4753-4761, 1997). For example, A2/K^b 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 ofthe 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 the peptide composition is a polypeptide, in PBS or Incomplete Freund's Adjuvant. Seven days after priming, splenocytes obtained from these animals are restimulated with syngenic iπadiated LPS-activated lymphoblasts coated with peptide.

Cell lines: Target cells for peptide-specific cytotoxicity assays are Jurkat cells transfected with the HLA- A2.1/K^b chimeric gene (e.g., Vitiello et al, J. Exp. Med. 173:1007, 1991) In vitro CTL activation: One week after priming, spleen cells (30xl0⁶ cells/flask) are co-cultured at 37°C with syngeneic, iπadiated (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 1.5xl0⁶) are incubated at 37°C in the presence of 200 μl of ⁵¹Cr. After 60 minutes, cells are washed three times and resuspended in R10 medium. Peptide is added where required at a concentration of 1 μg/ml. For the assay, 10^{4 5I}Cr-labeled target cells are added to different concentrations of effector cells (final volume of 200 μl) 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 and radioactivity is determined in a Micromedic automatic gamma counter. The percent specific lysis is determined by the formula: percent specific release = 100 x (experimental release - spontaneous release)/(maximum release - spontaneous release). To facilitate comparison between separate CTL assays run under the same conditions, % ⁵¹Cr 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 ⁵¹Cr release assay. To obtain specific lytic units/10⁶, the lytic units/10⁶ obtained in the absence of peptide is subtracted from the lytic units/10⁶ obtained in the presence of peptide. For example, if 30% ⁵¹Cr release is obtained at the effector (E): target (T) ratio of 50:1 (i.e., 5x10⁵ effector cells for 10,000 targets) in the 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.

The results are analyzed to assess the magnitude ofthe CTL responses of animals injected with the immunogenic CTL/HTL conjugate vaccine preparation and are compared to the magnitude ofthe 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 corifirm the immunogenicity of peptide conjugates containing multiple CTL epitopes and/or multiple HTL epitopes. In accordance with these procedures, it is found that a CTL response is induced, and concomitantly that an HTL response is induced upon administration of such compositions.

Example 20: Selection of CTL and HTL Epitopes for inclusion in a vaccine specific for a protein of Figure 2

This example illustrates a procedure for selecting peptide epitopes for vaccine compositions ofthe 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 ofthe following principles is balanced in order to make the selection.

Epitopes are selected which, upon administration, mimic immune responses that are coπelated with Figure 2 protein clearance. The number of epitopes used depends on observations of patients who spontaneously clear a Figure 2 protein. For example, if it has been observed that patients who spontaneously clear a Figure 2 protein generate an immune response to at least three (3) epitopes from a protein of Figure 2 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 IC₅₀ of 500 nM or less for an HLA class I molecule, or for class II, an IC₅₀ of 1000 nM or less; or HLA Class I peptides with high binding scores from the BIMAS web site, at URL bimas.dcrt.nih.gov/. In order to achieve broad coverage ofthe vaccine through out a diverse population, sufficient supermotif bearing peptides, or a sufficient aπay 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 the 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 the 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 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. Epitopes may be nested or overlapping (i.e., frame shifted relative to one another). For example, with overlapping epitopes, two 9-mer epitopes and one 10-mer epitope can be present in a 10 amino acid peptide. Each epitope can be exposed and bound by an HLA molecule upon administration of such a peptide. A multi-epitopic, peptide can be generated synthetically, recombinantly, or via cleavage from the native source. Alternatively, an analog can be made of this native sequence, whereby one or more ofthe epitopes comprise substitutions that alter the cross-reactivity and/or binding affinity properties ofthe polyepitopic peptide. Such a vaccine composition is administered for therapeutic or prophylactic purposes. This embodiment provides for the possibility that an as yet undiscovered aspect of immune system 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 the creating of any analogs) directs the immune response to multiple peptide sequences that are actually present in Figure 2 proteins, 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 that controls or clears cells that bear or overexpress a Figure 2 protein.

Example 21: Construction of "Minigene" Multi-Epitope DNA Plasmids

. This example discusses the construction of a minigene expression plasmid. Minigene plasmids may, of course, contain various configurations of B cell, CTL and/or HTL epitopes or epitope analogs as described herein.

A minigene expression plasmid typically includes multiple CTL and HTL peptide epitopes. In the present example, HLA-A2, -A3, -B7 supermotif-bearing peptide epitopes and HLA-A1 and -A24 motif-bearing peptide epitopes are used in conjunction with DR supermotif-bearing epitopes and or DR3 epitopes. HLA class I supermotif or motif-bearing peptide epitopes derived from a protein of Figure 2, are selected such that multiple supermotifs/motifs are represented to ensure broad population coverage. Similarly, HLA class II epitopes are selected from a Figure 2 protein to provide broad population coverage, i.e. both HLA DR- 1-4-7 supermotif-bearing epitopes and HLA DR-3 motif-bearing epitopes are selected for inclusion in the minigene construct. The selected CTL and HTL epitopes are then incorporated into a minigene for expression in an expression vector.

Such a construct may additionally include sequences that direct the HTL epitopes to the endoplasmic reticulum. For example, the Ii protein may be fused to one or more HTL epitopes as described in the art, wherein the CLIP sequence ofthe Ii protein is removed and replaced with an 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 plasmid. Other expression vectors that may be used for minigene compositions are available and known to those of skill in the art.

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

Overlapping oligonucleotides that can, for example, average about 70 nucleotides in length with 15 nucleotide overlaps, are synthesized and HPLC-purified. The oligonucleotides encode the selected peptide epitopes as well as appropriate linker nucleotides, Kozak sequence, and signal sequence. The final multiepitope minigene is assembled by extending the overlapping oligonucleotides in three sets, of reactions using PCR. A Perkin/Elmer 9600 PCR machine is used and a total of 30 cycles are performed using the following conditions: 95°C for 15 sec, annealing temperature (5° below the lowest calculated Tm of each primer pair) for 30 sec, and 72°C for 1 min.

For example, a minigene is prepared as follows. For a first PCR reaction, 5 μg 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 μl reactions containing Pfu polymerase buffer (lx= 10 mM KCL, 10 mM (NH4)₂S0₄, 20 mM Tris-chloride, pH 8.75, 2 mM MgS0₄, 0.1% Triton X-100, 100 μg/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 and 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 (Invifrogen) and individual clones are screened by sequencing.

Example 22: The Plasmid Construct and the Degree to Which It Induces Immunogenicity.

The degree to which a plasmid construct, for example a plasmid constructed in accordance with the previous Example, is able to induce immunogenicity is confirmed in vitro by determining epitope presentation by APC following fransduction or transfection ofthe APC with an epitope-expressing nucleic acid construct. Such a study determines "antigenicity" and allows the use of human APC. The assay determines the ability ofthe epitope to be presented by the APC in a context that is recognized by a T cell by quantifying the density of epitope-HLA class I complexes on the cell surface. Quantitation can be performed by directly measuring the amount of peptide eluted from the APC (see, 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 the amount of lysis or lymphokine release induced by diseased or transfected target cells, and then determining the concentration of peptide necessary to obtain equivalent levels of lysis or lymphokine release (see, e.g., Kageyama et al, J. Immunol. 154:567-576, 1995).

Alternatively, immunogenicity is confirmed through in vivo injections into mice and subsequent in vitro assessment of CTL and HTL activity, which are analyzed using cytotoxicity and proliferation assays, respectively, as detailed 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 supermotif peptide to induce CTLs in vivo, HLA-A2.1/K^b transgenic mice, for example, are immunized intramuscularly with 100 μg 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 ofthe respective compositions (peptide epitopes encoded in the minigene or the polyepitopic peptide), then assayed for peptide-specific cytotoxic activity in a ^5ICr release assay. The results indicate the magnitude ofthe CTL response directed against the A2- restricted epitope, thus indicating the in vivo immunogenicity ofthe minigene vaccine and polyepitopic vaccine.

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

To confirm the capacity of a class II epitope-encoding minigene to induce HTLs in vivo, DR transgenic mice, or for those epitopes that cross react with the appropriate mouse MHC molecule, I-A^b-restricted mice, for example, are immunized intramuscularly with 100 μg of plasmid DNA. As a means of comparing the level of HTLs induced by DNA immunization, a group of control animals is also immunized with an actual peptide composition emulsified in complete Freund's adjuvant. CD4+ T cells, i.e. HTLs, are purified from splenocytes of immunized animals and stimulated with each ofthe respective compositions (peptides encoded in the minigene). 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 the magnitude ofthe HTL response, thus demonstrating the in vivo immunogenicity ofthe minigene.

DNA minigenes, constructed as described in the 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., Bamett et al, Aids Res. and Human Retroviruses 14, Supplement 3:S299-S309, 1998) or recombinant vaccinia, for example, expressing a minigene or DNA encoding the complete protein of interest (see, e.g., Hanke et al, Vaccine 16:43.9-445, 1998; Sedegah et al, Proc. Natl. Acad. Sci USA 95:7648-53, 1998; Hanke and McMichael, Immunol. Letters 66:177-181, 1999; and Robinson et al, Nature Med. 5:526-34, 1999).

For example, the efficacy ofthe DNA minigene used in a prime boost protocol is initially evaluated in fransgenic mice. In this example, A2.1/K^b transgenic mice are immunized IM with 100 μg of a DNA minigene encoding the immunogenic peptides including at least one HLA-A2 supermotif-bearing peptide. After an incubation period (ranging from -3 -9 weeks), the mice are boosted IP with 10⁷ pfu/mouse of a recombinant vaccinia virus expressing the same sequence encoded by the DNA minigene. Control mice are immunized with 100 μg of DNA or recombinant vaccinia without the minigene sequence, or with DNA encoding the minigene, but without the vaccinia boost. After an additional incubation period of two weeks, splenocytes from the mice are immediately assayed for peptide-specific activity in an ELISPOT assay. Additionally, splenocytes are stimulated in vitro with the A2-restricted peptide epitopes encoded in the minigene and recombinant vaccinia, then assayed for peptide- specific activity in an alpha, beta and/or gamma IFN ELISA.

It is found that the minigene utilized in a prime-boost protocol elicits greater immune responses toward the HLA-A2 supermotif peptides than with DNA alone. Such an analysis can also be performed using HLA-A11 or HLA-B7 transgenic mouse models to assess CTL induction by HLA-A3 or HLA-B7 motif or supermotif epitopes. The use of prime boost protocols in humans is described below in the Example entitled "Induction of CTL Responses Using a Prime Boost Protocol ."

Example 23: Peptide Compositions for Prophylactic Uses

Vaccine compositions ofthe present invention can be used to prevent a gene of Figure 2 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% ofthe population, is aclministered to individuals at risk for a protein of Figure 2-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 μg, generally 100-5,000 μg, for a 70 kg patient. The initial adminisfration of vaccine is followed by booster dosages at 4 weeks followed by evaluation ofthe magnitude ofthe 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 protein of Figure 2-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 24: Polyepitopic Vaccine Compositions Derived from Native Protein Sequence of Figure 2

A native Figure 2protein sequence is analyzed, preferably using computer algorithms defined for each class I and/or class II supermotif or motif, to identify "relatively short" regions ofthe 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 the peptide, which coπesponds to the 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 ofthe 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, two 9-mer epitopes and one 10-mer 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 a protein antigen ofthe invention 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 ofthe epitopes comprise substitutions that alter the cross-reactivity and/or binding affinity properties ofthe polyepitopic peptide.

The embodiment of this example provides for the possibility that an as yet undiscovered aspect of immune system processing will apply to the native nested sequence and thereby facilitate the production of therapeutic or prophylactic immune response-inducing vaccine compositions. Additionally such an embodiment provides for the possibility of motif-bearing epitopes for an HLA makeup that is presently unknown. Furthermore, this embodiment (excluding an analoged embodiment) directs the immune response to multiple peptide sequences that are actually present in native proteins ofthe invention, thus avoiding the 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 the art which can be used to identify in a target sequence, the greatest number of epitopes per sequence length.

Example 25: Polyepitopic Vaccine Compositions from Multiple Antigens

The protein peptide epitopes ofthe present invention are used in conjunction with epitopes from other target tumor-associated antigens (such as from one or more proteins of Figure 2), to create a vaccine composition that is useful for the prevention or treatment of cancer that expresses protein(s) ofthe invention and such other antigens. For example, a vaccine composition can be provided as a single polypeptide that incorporates multiple epitopes from a protein ofthe invention as well as tumor-associated antigens that are often expressed witii the particular target cancer that is also associated with expression of a protein ofthe invention, or can be administered as a composition comprising a cocktail of one or more discrete epitopes. Alternatively, the vaccine can be administered as a minigene construct or as dendritic cells which have been loaded with the peptide epitopes in vitro.

Example 26: Use of peptides to evaluate an immune response

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

In this example highly sensitive human leukocyte antigen teframeric complexes ("tetramers") are used for a crossrsectional analysis of, for example, a protein of Figure 2 HLA-A*0201 -specific CTL frequencies from HLA A*0201-positive individuals at different stages of disease or following immunization comprising a protein of Figure 2 peptide containing an A*0201 motif. Teframeric complexes are synthesized as described (Musey et al, N. Engl. J. Med. 337:1267, 1997). Briefly, purified HLA heavy chain (A*0201 in this example) and β2-microglobulin are synthesized by means of a prokaryotic expression system. The heavy chain is modified by deletion ofthe ' fransmembrane-cytosolic tail and COOH-terminal addition of a sequence containing a BirA enzymatic biotinylation site. The heavy chain, β2-microglobulin, and peptide are refolded by dilution. The 45-kD refolded product is isolated by fast protein liquid chromatography and then biotinylated by BirA in the presence of biotin (Sigma, St. Louis, Missouri), adenosine 5' triphosphate and magnesium. Streptavidin-phycoerythrin conjugate is added in a 1:4 molar ratio, and the teframeric product is concentrated to 1 mg/ml. The resulting product is refeπed to as teframer- phycoerythrin.

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

Example 27: Use of Peptide Epitopes to Evaluate Recall Responses

The peptide epitopes ofthe 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 a protein of the invention-associated disease or who have been vaccinated with a protein ofthe invention vaccine.

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

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

In the microculture format, 4 x 10⁵ PBMC are stimulated with peptide in 8 replicate cultures in 96-well round bottom plate in 100 μl/well of complete RPMI. On days 3 and 10, 100 μl of complete RPMI and 20 U/ml final concenfration of rIL-2 are added to each well. On day 7 the cultures are fransfeπed into a 96-well flat-bottom plate and restimulated 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 ofthe eight replicate cultures to display greater than 10% specific ⁵¹Cr release, based on comparison with non-diseased control subjects as previously described (Rehermann, et al, Nature Med. 2: 1104, 1108, 1996; Rehermann et al, J. Clin. Invest. 97:1655-1665, 1996; and Rehermann et al. J. Clin. Invest. 98:1432-1440, 1996).

Target cell lines are autologous and allogeneic EBV-fransformed B-LCL that are either purchased from the American Society for Histocompatibility and Immunogenetics (ASHI, Boston, MA) or established from the pool of patients as described (Guilhot, et al. J. Virol. 66:2610-2618, 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 ofthe invention at 10 μM, and labeled with 100 μCi of ⁵¹Cr (Amersham Corp., Arlington Heights, IL) for 1 hour after which they are washed four times with HBSS.

Cytolytic activity is determined in a standard 4-h, split well ⁵¹Cr 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-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 proteins ofthe invention or a protein ofthe invention-related 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 μg/ml synthetic peptide ofthe invention, a whole protein ofthe invention antigens, 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 μCi 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-thymidine incoφoration in the presence of antigen divided by the ³H-thymidine incoφoration in the absence of antigen.

Example 28: Induction Of Specific CTL Response In Humans

A human clinical frial for an immunogenic composition comprising CTL and HTL epitopes ofthe invention is set up as an IND Phase I, dose escalation study and carried out as a randomized, double-blind, placebo- controlled trial. Such a trial is designed, for example, as follows:

A total of about 27 individuals are enrolled and divided into 3 groups:

Group I: 3 subjects are injected with placebo and 6 subjects are injected with 5 μg of peptide composition;

Group II: 3 subjects are injected with placebo and 6 subjects are injected with 50 μg peptide composition;

Group III: 3 subjects are injected with placebo and 6 subjects are injected with 500 μg 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 ofthe peptide composition as well as its immunogenicity. Cellular immune responses to the peptide composition are an index ofthe intrinsic activity of this the peptide composition, and can therefore be viewed as a measure of biological efficacy. The following summarize the clinical and laboratory data that relate to safety and efficacy endpoints.

Safety: The incidence of adverse events is monitored in the placebo and drag freatment 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 FicoU-Hypaque density gradient centrifugation, aliquoted in freezing media and stored frozen. Samples are assayed for CTL and HTL activity.

The vaccine is found to be both safe and efficacious.

Example 29: Phase II Trials In Patients Expressing a Gene of the Invention

Phase II trials are performed to study the effect of administering the CTL-HTL peptide compositions to patients having a cancer that expresses genes ofthe invention. The main objectives ofthe trial are to determine an effective dose and regimen for inducing CTLs in cancer patients that express a gene(s) ofthe invention, 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 ofthe same dose. The dosages are 50, 500 and 5,000 micrograms per injection. Drag- associated adverse effects (severity and reversibility) are recorded.

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

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 a gene ofthe invention-associated disease.

Example 30: Induction of CTL Responses Using a Prime Boost Protocol

A prime boost protocol similar in its underlying principle to that used to confirm the efficacy of a DNA vaccine in transgenic mice, such as described above in the Example entitled "The Plasmid Construct and the Degree to Which It Induces Immunogenicity," can also be used for the administration ofthe vaccine to humans. Such a vaccine regimen can include an initial adminisfration of, for example, naked DNA followed by a boost using recombinant virus encoding the vaccine, or recombinant protein/polypeptide or a peptide mixture administered in an adjuvant.

For example, the initial immunization may be performed using an expression vector, such as that constructed in the Example entitled "Construction of 'Minigene' Multi-Epitope DNA Plasmids" in the form of naked nucleic acid administered IM (or SC or ID) in the amounts of 0.5-5 mg at multiple sites. The nucleic acid (0.1 to 1000 μg) 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 viras administered at a dose of 5-10⁷ to 5x10⁹ pfu. An alternative recombinant virus, such as an MVA, canarypox, adenoviras, or adeno-associated viras, can also be used for the booster, or the polyepitopic protein or a mixture ofthe peptides can be administered. For evaluation of vaccine efficacy, patient blood samples are obtained before immunization as well as at intervals following adrninistration ofthe initial vaccine and booster doses ofthe vaccine. Peripheral blood mononuclear cells are isolated from fresh heparinized blood by FicoU-Hypaque density gradient centrifugation, aliquoted in freezing media and stored frozen. Samples are assayed for CTL and HTL activity.

Analysis ofthe results indicates that a magnitude of response sufficient to achieve a therapeutic or protective immunity against of Figure 2 is generated.

Example 31: Administration of Vaccine Compositions Using Dendritic Cells CDC)

Vaccines comprising peptide epitopes ofthe invention can be aclministered 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 ofthe invention. The dendritic cells are infused back into the patient to elicit CTL and HTL responses in vivo. The induced CTL and HTL then destroy or facilitate destruction, respectively, ofthe target cells that bear the proteins ofthe invention from which the epitopes in the vaccine are derived. For example, a cocktail of epitope-comprising peptides is administered ex vivo to PBMC, or isolated DC therefrom. A pharmaceutical to facilitate harvesting of DC can be used, such as Progenipoietin™ (Monsanto, St. Louis, MO) or GM-CSF/IL-4. After pulsing the DC with peptides, and prior to reinfusion into patients, the DC are washed to remove unbound peptides.

As appreciated clinically, and readily determined by one of skill based on clinical outcomes, the number of DC reinfused into the 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 10⁸ 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 ofthe DC. For example, PBMC generated after treatment with an agent such as Progenipoietin™ are injected into patients without purification ofthe DC. The total number of PBMC that are administered often ranges from 10⁸ to 10¹⁰. Generally, the cell doses injected into patients is based on the percentage of DC in the blood of each patient, as determined, for example, by immunofluorescence analysis wifli 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 the patient will be injected with a total of 2.5 x 10⁸ peptide-loaded PBMC. The percent DC mobilized by an agent such as 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 protein antigens ofthe invention can be induced by incubating, in tissue culture, the patient's, or genetically compatible, CTL or HTL precursor cells together with a source of APC, such as DC, and immunogenic peptides. After an appropriate incubation time (typically about 7-28 days), in which the precursor cells are activated and expanded into effector cells, the cells are infused into the patient, where they will destroy (CTL) or facilitate destruction (HTL) of their specific target cells, i.e., tumor cells.

Example 32: An Alternative Method of Identifying 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 the antigens of interest, e.g. antigens of Figure 2. Peptides produced by endogenous antigen processing of peptides produced as a result of transfection will then bind to HLA molecules within the cell and be transported and displayed on the cell's surface. Peptides are then eluted from the HLA molecules by exposure to mild acid conditions and their amino acid sequence deteimined, e.g., by mass spectral analysis (e.g., Kubo et al, J. Immunol. 152:3913, 1994). Because the majority of peptides that bind a particular HLA molecule are motif-bearing, this is an alternative modality for obtaining the motif-bearing peptides coπelated with the particular HLA molecule expressed on the cell.

Alternatively, cell lines that do not express endogenous HLA molecules can be transfected 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 proteins ofthe invention, to isolate peptides coπesponding to proteins of Figure 2 that have been presented on the cell surface. Peptides obtained from such an analysis will bear motif(s) that coπespond to binding to the single HLA allele that is expressed in the cell. As appreciated by one in the art, one can perform a similar analysis on a cell bearing more than one HLA allele and subsequently determine peptides specific for each HLA allele expressed. Moreover, one of skill would also recognize that means other than transfection, such as loading with a protein antigen, can be used to provide a source of antigen to the cell.

Example 33: Complementary Polynucleotides

Sequences complementary to Figure 2 protein-encoding sequences, or any parts thereof, are used to detect, decrease, or inhibit expression of naturally occurring proteins ofthe invention. Although use of oligonucleotides comprising from about 15 to 30 base pairs is described, essentially the 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 the coding sequences of proteins ofthe invention. To inhibit franscription, a complementary oligonucleotide is designed from the most unique 5' sequence and used to prevent promoter binding to the coding sequence. To inhibit translation, a complementary oligonucleotide is designed to prevent ribosomal binding to a Figure 2 protein-encoding franscript.

Example 34: Purification of Naturally-occurring or Recombinant Figure 2 Proteins Using Specific Antibodies

Naturally occurring or recombinant Figure 2 proteins are substantially purified by immunoaffinity chromatography using antibodies specific for a protein ofthe invention. An immunoaffinity column is constructed by covalently coupling, e.g., anti-protein of Figure 2 antibodies to an activated chromatographic resin, such as CNBr-activated SEPHAROSE (Amersham Pharmacia Biotech). After the coupling, the resin is blocked and washed according to the manufacturer's instructions.

Media containing protein(s) ofthe invention are passed over the immunoaffinity column, and the column is washed under conditions that allow the preferential absorbance of proteins ofthe invention (e.g., high ionic sfrength buffers in the presence of detergent). The column is eluted under conditions that disrupt antibody/Figure 2 protein binding (e.g., a buffer of pH 2 to pH 3, or a high concentration of a chaofrope, such as urea or thiocyanate ion), and GCR.P is collected.

Example 35: Identification of Molecules Which Interact with Proteins of the Invention

Figure 2 proteins, 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 aπayed in the wells of a multi-well plate are incubated with the labeled Figure 2 proteins, washed, and any wells with labeled Figure 2 protein complexes are assayed. Data obtained using different concentrations of Figure 2 proteins are used to calculate values for the number, affinity, and association of Figure 2 proteins with the candidate molecules.

Example 36: In Vivo Assay for Tumor Growth Promotion

The effect of a Figure 2 protein on tumor cell growth is evaluated in vivo by gene overexpression in tumor- bearing mice. For example, SCID mice are injected subcutaneously on each flank with 1 x 10⁶ of either PC3, DU145 or 3T3 cells containing tkNeo empty vector or a nucleic acid sequence ofthe invention. At least two strategies can be used: (1) Constitutive expression under regulation of a promoter such as a constitutive promoter obtained from the genomes of viruses such as polyoma viras, fowlpox virus (UK 2,211,504 published 5 July 1989), adenovirus (such as Adenoviras 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-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 the 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 the host cell systems. Tumor volume is then monitored at the appearance of palpable tumors and followed over time to determine if the cells expressing a gene ofthe invention grow at a faster rate and whether tumors of a Figure 2 protein-expressing cells demonstrate characteristics of altered aggressiveness (e.g. enhanced metastasis, vascularization, reduced responsiveness to chemotherapeutic drags).

Additionally, mice can be implanted with 1 x 10⁵ of the same cells orthotopically to determine if a protein ofthe invention has an effect on local growth in the prostate or on the ability ofthe cells to metastasize, specifically to lungs, lymph nodes, and bone maπow.

The assay is also useful to determine the inhibitory effect of candidate therapeutic compositions, such as for example, Figure 2 protein-related infrabodies, Figure 2 gene-related antisense molecules and ribozymes.

Example 37: Tumors In Vivo, With Monoclonals specific to a Figure 2 Protein

The significant expression of a Figure 2 proteins in cancer tissues of Table I and its restrictive expression in normal tissues, together with its expected cell surface expression, makes Figure 2 proteins excellent targets for antibody therapy. Similarly, Figure 2 proteins are a target for T cell-based immunotherapy. Thus, for Figure 2 genes expressed, e.g., in prostate cancer, the therapeutic efficacy of anti- Figure 2 protein mAbs in human prostate cancer xenograft mouse models is evaluated by using androgen-independent LAPC-4 and LAPC-9 xenografts (Craft, N., et al,. Cancer Res, 1999. 59(19): p. 5030-6) and the androgen independent recombinant cell line PC3-of Figure 2 (see, e.g., Kaighn, M.E., et al, Invest Urol, 1979. 17(1): p. 16-23); analogous models are used for other cancers.

Antibody efficacy on tumor growth and metastasis formation is studied, e.g., in a mouse orthotopic prostate cancer xenograft models and mouse kidney xenograft models. The antibodies can be unconjugated, as discussed in this Example, or can be conjugated to a therapeutic modality, as appreciated in the art. Anti-Figure 2 protein mAbs inhibit formation of both the androgen-dependent LAPC-9 and androgen-independent PC3-Figure 2 protein tumor xenografts. Anti-Figure 2 protein mAbs also retard the growth of established orthotopic tumors and prolonged survival of tumor-bearing mice. These results indicate the utility of anti-Figure 2 protein mAbs in the treatment of local and advanced stages of prostate cancer. (See, e.g., (Saffran, D., et al., PNAS 10:1073-1078 or World Wide Web URL www.pnas.org/cgi/doi/10.1073/pnas.051624698).

Administration ofthe anti-Figure 2 protein mAbs lead to retardation of established orthotopic tumor growth and inhibition of metastasis to distant sites, resulting in a significant prolongation in the survival of tumor- bearing mice. These studies indicate that proteins ofthe invention are attractive targets for immunotherapy and demonstrate the therapeutic potential of anti-Figure 2 protein mAbs for the freatment of local and metastatic cancer. This example demonstrates that unconjugated Figure 2 protein-related monoclonal antibodies are effective to inhibit the growth of human prostate tumor xenografts and human kidney xenografts grown in SCID mice; accordingly a combination of such efficacious monoclonal antibodies is also effective. Tumor inhibition using multiple unconjugated mAbs Materials and Methods

Figure 2 Protein-related Monoclonal Antibodies:

Monoclonal antibodies are raised against proteins ofthe invention as described in Example 10. The antibodies are characterized by ELISA, Western blot, FACS, and immunoprecipitation for their capacity to bind to the respective protein ofthe invention. Epitope mapping data for, e.g., the anti-Figure 2 protein mAbs, as determined by ELISA and Western analysis, indicate that the antibodies recognize epitopes on the respective Figure 2 protein. Immunohistochemical analysis of prostate cancer tissues and cells with these antibodies is performed.

The monoclonal antibodies are purified from ascites or hybridoma tissue culture supernatants 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 and used for the treatment of mice receiving subcutaneous or orthotopic injections of LAPC-9 prostate tumor xenografts.

Cancer Xenografts 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 immunodeficient (SCID) mice (Taconic Farms) by s.c. trocar implant (Craft, N., et al, supra). The prostate carcinoma cell line PC3 (American Type Culture Collection) is maintained in RPMI supplemented with L-glutamine and 10% FBS.

Recombinant PC3 and 3T3- cell populations expressing a protein ofthe invention are generated by retroviral gene fransfer 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): p. 14523-8. Anti-protein ofthe invention staining is detected by using an FITC-conjugated goat anti-mouse antibody (Southern Biotechnology Associates) followed by analysis on a Coulter Epics-XL flow cytometer.

Xenograft Mouse Models.

Subcutaneous (s.c.) tumors are generated by injection of 1 x 10 ⁶ LAPC-9, PC3, recombinant PC3-protein ofthe invention, 3T3 or recombinant 3T3-protein ofthe invention cells mixed at a 1 :1 dilution with Matrigel (Collaborative Research) in the right flank of male SCID mice. To test antibody efficacy on tumor formation, i.p. antibody injections are started on the same day as tumor-cell injections. As a confrol, mice are injected with either purified mouse IgG (ICN) or PBS; or a purified monoclonal antibody that recognizes an irrelevant antigen not expressed in human cells. In preliminary studies, no difference is found between mouse IgG or PBS on tumor growth. Tumor sizes are determined by vernier caliper measurements, and the tumor volume is calculated as length x width x height. Mice with s.c. tumors greater than 1.5 cm in diameter are sacrificed. PSA levels are determined by using a PSA ELISA kit (Anogen, Mississauga, Ontario). Circulating levels of, e.g., anti-Figure 2 protein mAbs are determined by a capture ELISA kit (Bethyl Laboratories, Montgomery, TX). (See, e.g., Saffran, D., et al., PNAS 10:1073-1078 or www.pnas.org/cgi/ doi/10.1073/pnas.051624698)

Orthotopic injections are performed under anesthesia by using ketamine/xylazine. For prostate orthotopic studies, an incision is made through the abdominal muscles to expose the bladder and seminal vesicles, which then are delivered through the incision to expose the dorsal prostate. LAPC-9 or PC3 cells (5 x 10^s ) mixed with Matrigel are injected into each dorsal lobe in a 10-μl volume. To monitor tumor growth, mice are bled on a weekly basis for determination of PSA levels. The mice are segregated into groups for the appropriate treatments, with anti-protein ofthe invention or confrol mAbs being injected i.p. Anti-Figure 2 Protein mAbs Inhibit Growth of Respective Figure 2 Protein-Expressing Xenograft-Cancer Tumors

The effect of anti-Figure 2 protein mAbs on tumor formation is tested by using LAPC-9 and recombinant PC3-protein ofthe invention orthotopic models. As compared with the s.c. tumor model, the orthotopic model, which requires injection of tumor cells directly in the mouse prostate or kidney, respectively, results in a local tumor growth, development of metastasis in distal sites, deterioration of mouse health, and subsequent death (Saffran, D., et al., PNAS supra; Fu, X., et al., Int J Cancer, 1992. 52(6): p. 987-90; Kubota, T., J Cell Biochem, 1994. 56(1): p. 4-8). The features make the orthotopic model more representative of human disease progression and allowed us to follow the therapeutic effect of mAbs on clinically relevant end points.

Accordingly, tumor cells are injected into the mouse prostate or kidney, and 2 days later, the mice are segregated into two groups and treated with either: a) 200-500μg, of anti-Figure 2 protein Ab, or b) PBS three times per week for two to five weeks.

A major advantage ofthe orthotopic prostate-cancer model is the ability to study the development of metastases. Formation of metastasis in mice bearing established orthotopic tumors is studied by IHC analysis on lung sections using an antibody against a prostate-specific cell-surface protein STEAP expressed at high levels in LAPC-9 xenografts (Hubert, R.S., et al, Proc Natl Acad Sci U S A, 1999. 96(25): p. 14523-8).

Mice bearing established orthotopic LAPC-9 or recombinant PC3-Figure 2 protein tumors are administered lOOOμg injections of either anti-Figure 2 protein mAbs or PBS over a 4-week period. Mice in both groups are allowed to establish a high tumor burden (PSA levels greater than 300 ng/ml for 1APC-9), to ensure a high frequency of metastasis formation in mouse lungs. Mice then are killed and their prostate and lungs are analyzed for the presence of tumor cells by IHC analysis.

These studies demonsfrate a broad anti-tumor efficacy of anti-Figure 2 protein antibodies on initiation and progression of prostate cancer in xenograft mouse models. Anti-Figure 2 protein antibodies inhibit tumor formation of both androgen-dependent and androgen-independent tumors, retard the growth of aheady established tumors, and prolong the survival of treated mice. Moreover, anti-Figure 2 protein mAbs demonsfrate a dramatic inhibitory effect on the spread of local prostate tumor to distal sites, even in the presence of a large tumor burden. Thus, anti- Figure 2 protein mAbs are efficacious on major clinically relevant end points (tumor growth), prolongation of survival, and health.

Example 38: Therapeutic and Diagnostic use of Antibodies specific to a protein of Figure 2

Anti-protein of Figure 2 monoclonal antibodies are safely and effectively used for diagnostic, prophylactic, prognostic and/or therapeutic puφoses in humans. Western blot and immunohistochemical analysis of cancer tissues and cancer xenografts with anti-protein of Figure 2 mAb show strong extensive staining in carcinoma but significantly lower or undetectable levels in normal tissues. Detection of a protein of Figure 2 in carcinoma and in metastatic disease demonsfrates the usefulness ofthe mAb as a diagnostic and/or prognostic indicator. Anti-protein of Figure 2 antibodies are therefore used in diagnostic applications such as immunohistochemistry of biopsy specimens to detect cancer from suspect patients.

As determined by flow cytometry, anti-protein of Figure 2 mAbs specifically bind to carcinoma cells. Thus, anti-protein of Figure 2 antibodies are used in diagnostic whole body imaging applications, such as radioimmunoscintigraphy and radioimmunotherapy, (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 a protein of Figure 2. Shedding or release of an extracellular domain of a protein of Figure 2 into the extracellular milieu, such as that seen for alkaline phosphodiesterase BIO (Meerson, N. R., Hepatology 27:563-568 (1998)), allows diagnostic detection of a protein of Figure 2 by coπesponding anti-protein of Figure 2 antibodies in serum and/or urine samples from suspect patients.

Anti-protein of Figure 2 antibodies that specifically bind protein of Figure 2 are used in therapeutic applications for the treatment of cancers that express that protein of Figure 2. Anti-protein of Figure 2 antibodies are used as an unconjugated modality and as a conjugated form in which the antibodies are attached to one of various therapeutic or imaging modalities well known in the art, such as a prodrugs, enzymes or radioisotopes. In preclinical studies, unconjugated and conjugated anti-protein of Figure 2 antibodies are tested for efficacy of tumor prevention and growth inhibition in the SCID mouse cancer xenograft models, e.g., kidney cancer models AGS-K3 and AGS-K6, (see, e.g.,, the Example entitled "Monoclonal Antibody-mediated Inhibition of Prostate Tumors In vivo"). Conjugated and unconjugated anti-protein of Figure 2 antibodies are used as a therapeutic modality in human clinical trials either alone or in combination with other treatments as described in the following Examples.

Example 39: Human Clinical Trials for the Treatment and Diagnosis of Human Carcinomas through use of Human Antibodies Specific for a Protein of Figure 2 In vivo

Antibodies are used in accordance with the present invention which recognize an epitope of a Figure 2 protein, and are used in the treatment of certain tumors such as those listed in Table I. Based upon a number of factors, including Figure 2 protein expression levels, tumors such as those listed in Table I are presently prefeπed indications. In connection with each of these indications, three clinical approaches are successfully pursued.

I.) Adjunctive therapy: In adjunctive therapy, patients are treated with antibodies ofthe invention, e.g., antibodies that specifically bind a protein ofthe invention, in combination with a chemotherapeutic or antineoplastic agent and or radiation therapy. Primary cancer targets, such as those listed in Table I, are freated under standard protocols by the addition anti-Figure 2 protein antibodies to standard first and second line therapy. Protocol designs address effectiveness as assessed by reduction in tumor mass as well as the ability to reduce usual doses of standard chemotherapy. These dosage reductions allow additional and/or prolonged therapy by reducing dose-related toxicity ofthe chemotherapeutic agent. Anti-Figure 2 protein 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 doxorabicin (preclinical).

II.) Monotherapy: In connection with the use ofthe anti-Figure 2 protein 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-Figure 2 protein antibodies, the radiolabeled antibodies are utilized as a diagnostic and/or imaging agent. In such a role, the labeled antibodies localize to both solid tumors, as well as, metastatic lesions of cells expressing a protein ofthe invention. In connection with the use ofthe anti-Figure 2 protein antibodies as imaging agents, the antibodies are used as an adjunct to surgical treatment of solid tumors, as both a pre-surgical screen as well as a post-operative follow-up to determine what tumor remains and/or returns. In one embodiment, a (" ' In)-Figure 2 protein antibody is used as an imaging agent in a Phase I human clinical trial in patients having a carcinoma that expresses a protein of the invention (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

As appreciated by those of ordinary skill in the art, dosing considerations can be determined through comparison with the analogous products that are in the clinic. Thus, anti-Figure 2 protein 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-Figure 2 protein 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-Figure 2 protein antibodies tiiat are fully human antibodies, as compared to the chimeric antibody, have slower clearance; accordingly, dosing in patients with such fully human anti- Figure 2 protein 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 delivery of anti-Figure 2 protein antibodies. Conventional intravenous delivery is one standard delivery technique for many tumors. However, in connection with tumors in the peritoneal cavity, such as tumors ofthe ovaries, biliary duct, other ducts, and the like, infraperitoneal adminisfration may prove favorable for obtaining high dose of antibody at the tumor and to also minimize antibody clearance. In a similar manner, certain solid tumors possess vasculature that is appropriate for regional perfusion. Regional perfusion allows for a high dose of antibody 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-Figure 2 protein antibodies in connection with adjunctive therapy, monotherapy, and as an imaging agent. Trials initially demonstrate safety and thereafter confirm efficacy in repeat doses. Trials are open label comparing standard chemotherapy with standard therapy plus anti-Figure 2 protein antibodies. As will be appreciated, one criteria that can be utilized in connection with enrollment of patients is Figure 2 protein expression levels in their tumors as determined e.g. from biopsy specimens. 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 a protein ofthe invention. Standard tests and follow-ups are utilized to monitor each of these safety concerns. Anti-Figure 2 protein antibodies are found to be safe upon human administration.

Example 40: Human Clinical Trial Adjunctive Therapy with Human Antibody (Specific to a Protein of Figure 2. and Chemotherapeutic Agent

A phase I human clinical trial is initiated to assess the safety of six infravenous doses of a human anti- Figure 2 protein antibody in connection with the treatment of a solid tumor, e.g., a cancer of a tissue listed in Table I. In the study, the safety of single doses of anti-Figure 2 protein antibodies when utilized as an adjunctive therapy to an antineoplastic, or chemotherapeutic agent, such as cisplatin, topotecan, doxorabicin, adriamycin, taxol, or the like,' is assessed. The trial design includes delivery of six single doses of an anti-Figure 2 protein antibody with dosage of antibody escalating from approximately about 25 mg m ² to about 275 mg/m ² over the course of the freatment in accordance with the following schedule: Day O Day 7 Day 14 Day 21 Day 28 Day 35

mAb Dose 25 75 125 175 225 275 mg/m² mg/m ² mg/m ² mg/m ² mg/m² mg/m² Chemotherapy + + + + + +

(standard dose)

Patients are closely followed for one-week following each administration of antibody and chemotherapy. In particular, patients are assessed for the safety concerns mentioned above: (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 human antibody therapeutic, or HAHA response); and, (iii) toxicity to normal cells that express a protein ofthe invention. Standard tests and follow-up are utilized to monitor each of these safety concerns. Patients are also assessed for clinical outcome, and particularly reduction in tumor mass as evidenced by MRI or oflier imaging.

The anti-Figure 2 protein antibodies are demonstrated to be safe and efficacious, Phase II trials confirm the efficacy and refine optimum dosing.

Example 41: Human Clinical Trial: Monotherapy with Human Antibody Specific to a Protein of

Figure 2 ■ Anti-Figure 2 protein antibodies are safe in connection with 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 the above-described adjunctive trial with the exception being that patients do not receive chemotherapy concuπently with the receipt of doses of anti-Figure 2 protein antibodies.

Example 42: Human Clinical Trial: Diagnostic Imaging with Antibody Specific to a Protein of

Figure 2

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-Figure 2 protein 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 et 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 43: Effects on Tumor Growth and Promotion

The genes in Figure 2 contribute to the growth of cancer cells. The role of these genes in tumor growth is investigated in a variety of primary and fransfected cell lines including prostate, colon, bladder and kidney cell lines as well as NIH 3T3 cells engineered to stably express the gene of interest. Parental cells lacking the gene of interest and cells expressing that gene 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 Drags. 1996, 7:288). To determine the role of genes in Figure 2 in the transformation process, the effect of individual genes in colony forming assays is investigated. Parental NIH3T3 cells lacking the gene of interest are compared to NHI-3T3 cells expressing that gene, using a soft agar assay under stringent and more permissive conditions (Song Z. et al. Cancer Res. 2000;60:6730). It is found that genes set forth in Figure 2 asversely affect fransformation.

To determine the role ofthe genes of Figure 2 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 the gene of interest are compared to cells expressing that gene. Cells are loaded with the fluorescent dye, calcein, and plated in the top well ofthe Transwell insert coated with a basement membrane analog. Invasion is determined by fluorescence of cells in the lower chamber relative to the fluorescence ofthe entire cell population. It is found that genes set forth in Figure 2 adversely invasion and/or metastasis.

The genes in Figure 2 also play a role in cell cycle modulation and apoptosis. Parental cells and cells expressing the gene of interest 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 and stained with anti-BrdU Ab and propidium iodide. Cells are analyzed for entry into the GI, S, and G2M phases ofthe cell cycle. Alternatively, the effect of stress on apoptosis is evaluated in control parental cells and cells expressing the gene of interest, including normal and tumor prostate, colon and lung cells. Engineered and parental cells are treated with various chemotherapeutic agents, such as etoposide, flutamide, etc, and protein synthesis inhibitors, such as cycloheximide. Cells are stained with annexin V-FITC and cell death is measured by FACS analysis. The modulation of cell death by genes of Figure 2 play a critical role in regulating tumor progression and tumor load.

When a genes set for in Figure 2, and/or its respective gene product, plays a role in cell growth, transformation, invasion or apoptosis, it is used as a target for diagnostic, prognostic, preventative and therapeutic puφoses.

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 disclosures of each of these references are hereby incoφorated by reference herein in their entireties.

The present invention is not to be limited in scope by the embodiments disclosed herein, which are intended as single illustrations of individual aspects ofthe invention, and any that are functionally equivalent are within the scope ofthe invention. Various modifications to the models and methods ofthe invention, in addition to those described herein, will become apparent to those skilled in the art from the foregoing description and teachings, and are similarly intended to fall within the scope ofthe invention. Such modifications or other embodiments can be practiced without departing from the true scope and spirit ofthe invention. TABLES

TABLE I: Exemplary Tissues that Express a Gene of Figure 2 When Malignant.

TABLE II: AMINO ACID ABBREVIATIONS

TABLE III: AMINO ACID SUBSTITUTION MATRIX

Adapted from the GCG Software 9.0 BLOSUM62 amino acid substitution matrix (block substitution matrix). The higher the value, the more likely a substitution is found in related, natural proteins. (See URL www.ikp.unibe.ch/manual/blosum62.htinl )

A C D E F G H I K L M N P Q R S T V W y .

4 0 -2 -1 -2 0 -2 -1 -1 -1 -1 -2 -1 -1 -1 1 0 0 -3 -2 A

9 -3 -4 -2 -3 -3 -1 -3 -1 -1 -3 -3 -3 -3 -1 -1 -1 -2 -2 C

6 2 -3 -1 -1 -3 -1 -4 -3 1 -1 0 -2 0 -1 -3 -4 -3 D

5 -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 1

5 -2 -1 0 -1 1 2 0 -1 -2 -3 -2 K

4 2 -3 -3 -2 -2 -2 -1 1 -2 -1 L

5 -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

5 1 0 -1 -2 -2 -1 Q

5 -1 -1 -3 -3 -2 R

4 1 -2 -3 -2 S

5 0 -2 -2 T

4 -3 -1 V

11 2 W

7 Y

TABLE IV (A) HLA Class I Supermotifs/Motifs

Bolded residues are prefeπed, italicized residues are less prefeπed: 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 H SUPERMOTIF

TABLE IV (C): HLA Class π Motifs

MOTIFS 1° anchor 1 2 3 4 5 1° anchor 6 7 8 9

DR4 prefeπed FMYLIVW M T I YSTCPALIM MH MH deleterious W R WDE

DRl prefeπed MFLIVWY PAMQ VMATSPLIC M AVM deleterious C CH FD CWD GDE D

DR7 prefeπed MFLIVWY M W A TVMSACTPL M IV deleterious C G GRD N G

DR3 MOTIFS 1° anchor 1 1° anchor 4 1° anchor 6 motif a LΓVMFY D prefeπed motif b LΓVMFAY DNQEST KRH prefeπed

DR MFLIVWY VMSTACPI/

Supermotif

Italicized residues indicate less prefeπed or "tolerated" residues

TABLE IV (D): HLA Class I Supermotifs

POSITION: C-terminus

SUPERMOTIFS

Al 1° Anchor 1° Anchor TIL VMS FWY

A2 1° Anchor 1° Anchor UVMATQ LΓVMAT

A3 prefeπed 1° Anchor YFW YFW YFW P T Anchor YSMATLI (4/5) (3/5) (4/5) (4/5) RK deleterious DE (3/5); DE P (5/5) (4/5)

A24 1° Anchor 1° Anchor YFWIVLMT FTYWLM

B7 prefeπed FWY (5/5) 1° Anchor FWY FWY l°Anchor

LrVM (3/5) P (4/5) (3/5) YTLFMWYA deleterious DE (3/5); DE G QN DE P(5/5); (3/5) (4/5) (4/5) (4/5) ⁽3(4/5); A(3/5); QN(3/5)

B27 1° Anchor 1 "Anchor RHK FYLWMIVA

B44 1° Anchor 1° Anchor ED FWYLΓMVA

■ B58 1° Anchor 1° Anchor ATS FWYLIVMA

B62 1° Anchor 1° Anchor QUVMP FWYMIVLA

Itahcized residues indicate less prefeπed or "tolerated" residues

TABLE IV (E): HLA Class I Motifs

POSITION: C-teπriinus or

C-terminus

Al prefeπed GFY TAnchor DEA YFW DEQN YFW TAnchor 9-mer W STM Y deleterious DE RHKLIVMP G

Al prefeπed GRHK ASTCLIVM TAnchor GSTC ASTC LIVM DE TAnchor

9-mer DE_4S Y deleterious A RHKDEPY DE PQN RHK PG GP

FW

Al prefeπed YFW TAnchor DEAQN A YFWQN PASTC GDE P TAnchor

10-mer STM Y deleterious GP RHKGLrVM DE RHK QNA RHKYFW RHK A

Al prefeπed YFW STCLΓVM TAnchor A YFW PG G YFW TAnchor

10-mer ΏΈAS Y deleterious RHK RHKDEPY P G PRHK QN

FW

A2.1 prefeπed YFW TAnchor YFW STC YFW A . P TAnchor

9-mer LMIVQAT YLIMAT deleterious DEP DERKH RKH DERKH

Italicized residues indicate less prefeπed or "tolerated" residues

TABLE TV (E): HLA Class I Motifs, continued

POSITION: 1 2 3 4 5 6 7 8 9 C-Terminus

A2.1 prefeπed AYFW TAnchor LVIM G G FYWL TAnchor

10-mer IMIVQA VIM YLIMAT T deleterious DEP DE RKHA P RKH DERKH RKH

A3 prefeπed RHK TAnchor YFW PRHKYFW A YFW P TAnchor

LMVISA KYRHFA

TFCGD deleterious DEP DE

All prefeπed A TAnchor YFW YFW A YFW YFW P TAnchor VTLMIS KRYH AGNCD deleterious DEP A G

A24 prefeπed YFWRHK TAnchor STC YFW YFW TAnchor

9-mer YFWM FLIW deleterious DEG DE G QNP DERH G AQN K

A24 prefeπed TAnchor P YFWP P TAnchor

10-mer YFWM FLIW deleterious GDE QN RHK DE A QN DEA

A3101 prefeπed RHK TAnchor YFW P YFW YFW AP TAnchor MVTALIS RK deleterious DEP DE ADE DE DE DE

A3301 prefeπed TAnchor YFW AYFW TAnchor MVALF7 RK

ST deleterious GP DE

Italicized residues indicate less prefeπed or "tolerated" residues

TABLE IV (E): HLA Class I Motifs, continued

POSITION 1 2 3 4 ^" 5 6 7 8 9 C-Terminus

A6801 prefeπed YFWSTC TAnchor YFWLIV YFW P TAnchor AYTMSLI M RK deleterious GP DEG RHK A

B0702 prefeπed RHKFW TAnchor RHK RHK RHK RHK PA TAnchor

Y P MFWYAIV deleterious DEQNP DEP DE DE GDE QN DE

B3501 prefeπed FWYLΓV TAnchor FWY FWY TAnchor

M P LMFWY/K

A deleterious AGP G G

B51 prefeπed LΓVMFW TAnchor FWY STC FWY G FWY TAnchor Y P VFWYAM deleterious AGPDER DE G DEQN GDE HKSTC

B5301 prefeπed LIVMFW TAnchor FWY STC FWY LΓVMFWY FWY TAnchor Y P 1MFWY_1__ V deleterious AGPQN G RHKQN DE

B5401 prefeπed FWY TAnchor FWYL LIVM ALIVM FWYAP TAnchor P JVM ATΓVIMF

WY deleterious GPQNDE GOES RHKDE DE QNDGE DE TC

Italicized residues indicate less prefeπed or "tolerated" residues. The information in this Table is specific for 9-mers unless otherwise specified.

TABLE XVI - 187P3F2 HLA B7 10-MERS

SEQ. ID

START SUBSEQUENCE SCORE NUM.

TABLE XI 192P2G7 HLA AllOl 9- MERS

SEQ. ID

START SUBSEQUENCE SCORE NUM.

267 LVYKQKMGK 8.000

24 GVRLPPFCR 3.600

47 DVWIVTYPK 2.400

139 WSYYQFHR 1.200

156 GTFQEFCRR 1.200

10 STPGEFESK 1.000

212 RFLGVSCDK 0.906

128 VIYMARNPK 0.800

204 VTMVEQLAR 0.800

18 KYFEFHGVR 0.480

178 HVQEFWEHR 0.400

I 264 KFDLVYKQK 0.300

101 ELTSPRLIK 0.240

256 IFTVSMNEK 0.200

161 FCRRFMNDK 0.200

193 FLKYEDMHR ||0.160 246 GRGRVGLWK 0.120

TABLE XIII - 192P2G7 HLA A24 MERS

SEQ. ID

START SUBSEQUENCE SCORE NUM.

195 KYEDMHRDL 720.000

52 TYPKSGTSL 300.000

141 SYYQFHRSL 200.000

153 SYRGTFQEF 158.400

90 EYPQPGLDI 75.000

19 YFEFHGVRL 30.000

113 PYRFLPSDL 24.000

157 TFQEFCRRF 18.000

14 EFESKYFEF 16.500

78 GLMNIDEQL 8.400

186 RMDSNVLFL 8.000

270 KQKMGKCDL 8.000

109 KSHLPYRFL 8.000

206 MVEQLARFL 7.200

150 RTMSYRGTF 6.000

59 SLLQΞWYL 6.000

259 VSMNEKFDL 6.000

171 GYGSWFEHV 6.000

88 VLEYPQPGL 6.000

202 DLVTMVEQL 6.000

104 SPRLIKSHL 5.600

268 VYKQKMGKC 5.500

81 NIDEQLPVL 4.800

225 ALTEHCHQL 4.800

71 GADPDEIGL 4.800

272 KMGKCDLTF 4.000

235 DQCCNAEAL 4.000

215 GVSCDKAQL 4.000

53 YPKSGTSLL 4.000

249 RVGLWKDIF 4.000

130 YMARNPKDL 4.000

205 TMVEQLARF 3.600

257 FTVSMNEKF 3.300

TABLE XX; Motif-bearing Subsequences of a Protein of Figure 2

74P3B3 var . l

Tyrosine sulfation site

91 - 105 fqseneaYppaeris

Protein kinase C phosphorylation site

127 - 129 TdK

166 - 168 SsR

176 - 178 SgR

Casein kinase II phosphorylation site

93 - 96 SenE

105 - 108 SaeE

119 - 122 SeeD

136 - 139 SfeE

N-myristoylation site

2 - 7 GQskSK

23 - 28 GlkaST

109 - 114 GGdaAE

115 - 120 GGedSE

172 - 177 GlirSG

74P3B3 var.2

Protein kinase C phosphorylation site 4 - 6 TkK 116 - 118 SfK Casein kinase II phosphorylation site

151 - 154 TpcD N-myristoylation site

106 - 111 GAhgAV

204 - 209 GGq GT

205 - 210 GQ gTV 208 - 213 GTvnNH

83P4B8

N-glycosylation site

580 - 583 NETF

740 - 743 NISA

757 - 760 NPSI

798 - 801 NKTS

1218 - 1221 NKSK

1224 - 1227 NYTG

Tyrosine sulfation site

692 - 706 eeeeeafYedlddil cAMP- and cGMP-dependent protein kinase phosphorylation site 614 - 617 RRnS 779 - 782 KKlS Protein kinase C phosphorylation site 23 - 25 TlR

62 - 64 TlR

139 - 141 TkK

166 - 168 SgR

221 - 223 SsK

225 - 227 SrK

337 - 339 SfK

429 - 431 TfK

478 - 480 SsK 512 - 514 SmR

590 - 592 SIR

710 - 712 TnR

806 - 808 SmK

895 - 897 SgK

937 - 939 TdK

952 - 954 TqR

1010 - 1012 TsK

1038 - 1040 SyK

1268 - 1270 SkK

1283 - 1285 TsR

Casein kinase II phosphorylation site

23 - 26 TlrE

139 - 142 TkkE

152 - 155 SgeE

187 - 190 TaeE

228 - 231 SvlE

238 - 241 SalD

249 - 252 SgdE

337 - 340 SfkD

512 - 515 SmrD

653 - 656 TqgD

659 - 662 SlqE

716 - 719 SelE

824 - 827 ShqE

834 - 837 SsnE

860 - 863 SgpD

890 - 893 TsvE

891 - 894 SveE

937 - 940 TdkE

971 - 974 SsqE

972 - 975 SqeE

1018 - 1021 SreD

1090 - 1093 SqaE

1114 - 1117 TlsE

.1157 - 1160 ScvD

1283 - 1286 TsrD

1310 - 1313 TasE Tyrosine kinase phosphorylation site

480 - 487 Kvt . EafdY

833 - 841 RssnEfmrY

N-myristoylation site

53 - 58 GSpcSE

119 - 124 GSlvNG

149 - 154 GVlsGE

224 - 229 GSrkSV

551 - 556 GSlsSS

736 - 741 GlknNI

1139 - 1144 GT11TF

1189 - 1194 GIpkNM

Amidation site

895 - 898 sGKK

Bipartite nuclear targeting sequence

779 - 795 KKlsdilnekagkaktk ATP/GTP-binding site motif A (P-loop)

119 - 126 GslvnGKS

896 - 903 GkkekGKS Leucine zipper pattern

130 - 151 LpiiltaLatkken aygkgv

109P1D4

N-glycosylation :site

27 - 30 NYTI

48 - 51 N SL

54 - 57 NKSL

145 - 148 NISI

303 - 306 NATT

344 - 347 NVTD

365 - 368 NDTV

447 - 450 NQSA

553 - 556 NVTV

628 - 631 NISF

679 - 682 NCSY

703 - 706 NDTG

732 - 735 NITL

773 - 776 NESV

778 - 781 NATL

1018 - 1021 NTTF Tyrosine sulfation site

416 - 430 flletaaYldyestk

419 - 433 etaayldYestkeya

592 - 606 itvtdpdYgdnsavt cAMP- and cGMP-dependent protein kinase phosphorylation site

787 - 790 RKsT

874 - 877 KKhS Protein kinase C phosphorylation site 29 - 31 TiR

310 - 312 TiK

386 - 388 TdK

428 - 430 StK

656 - 658 SaK

877 - 879 SpK

927 - 929 TfK

Casein kinase II phosphorylation site

147 - 150 SipE

238 - 241 SvtD

252 - 255 TeiE

257 - 260 SipE

274 - 277 TdaD

310 - 313 TikE

384 - 387 TvtD

386 - 389 TdkD

400 - 403 Td E

428 - 431 StkE

474 - 477 SipE

489 - 492 SatriD

559 - 562 SiiD

571 - 574 ThnE

593 - 596 TvtD

595 - 598 TdpD

608 - 611 SilD

734 - 737 TlmE 887 - 890 TieE

891 - 894 TkaD

984 - 987 SssD

990 - 993 SvsD

99 - 1002 TtfE

Tyrosine kinase phosphorylation site 530 - 536 Rek . Edk . Y N-myristoylation site

265 - 270 GTsvTQ

333 - 338 GLmpAR

482 - 487 GlqlTK

648 - 653 GGrvSR

757 - 762 GQpdSL

900 - 905 GNrvTL

1013 - ■ 1018 GlqvSN

Cadherin domain signature

237 - 247 VsVtDtNDNhP

343 - 353 VnVtDvNDNvP

454 - 464 IkVkDeNDNaP

558 - 568 VsIiDqNDNsP

661 - 671 InVvDvNDNkP Cadherins domain

140 - ₂49 PATVINISIPENSAINSKY..TLPAAVDPDVGINGVQNYELIKSQ---..NIFGLDVIEt pegdkmPQLIVQKELDRE EKDTYVMKVKVEDGG. FPQRSSTAI QVSVTDT

NDN HPVF

26 - 139 KNYTIREEMPENVLIGDL kdLNLSLIPN -SLTTAMQFK VYKTGDV.. PLIRIEEDTg EIFTTGARIDREklcagipr EHCFYEVEVAILPDE. IFRLVKIRFUEDI

NDN APLF

250 - 355 KETEIEVSIPENAPVGTSV. .TQLHATDADIGENAKIHFSFSNLVSNIarR FH NATT . GLITIKEPLDRE ETPNHKL VI ASDGG . - -LMPARAMVLVNVTDV

NDN VPSI

362 - 466 NPVNDTW SENIPLNTKI ..ALITVTDKDADHNGRVTCFTDHEI- -- ..-PFR RPVFs n.... qFLLETAAYLDYE STKEYAIKLLAADAG . KPPLNQSAMLFIKVKDE

NDN APVF

467 - 570 TQSFVTVSIPENNSPGIQL. . TKVSA DADSGPNAKINYLLGPDAPP- . . -EFS DCRT. GMLTWKKLDRE KEDKYLFTILAKDNG . VPPLTSNVTVFVSIIDQ

NDN SPVF

571 - 673 THNEYNFYVPENLPRHGTV. .GLITVTDPDYGDNSAVT SILDEND-- . . -DFTIDSQT. GVIRPNISFDRE KQESYTFYVKAEDGG . RVSRSSSAKVTINWDV

NDN KPVF

685 - 795 VLPSTNPGTW.. QVIAVDNDTGMNAEVCYSIVGGNTRD ..- FAIDQET. GNITLMEKCDVT DLGLHRVLVKANDLGqPDS FSWIVNLFVNES

VTNat1inelvrksteAPVT

151P1C7A

N-glycosylation site

256 - 259 NSSR cAMP- and cGMP-dependent protein kinase phosphorylation site

170 - 173 RRtT

225 - 228 RKgS Protein kinase C phosphorylation site

169 - 171 SrR

175 - 177 SsK

244 - 246 ScR

257 - 259 SsR Casein kinase II phosphorylation site

153 - 156 TitE 163 - 166 StlD N-myristoylation site

5 - 10 GAagAT 28 ^■- 33 GVsaTL

50 - 55 GGaaGH

57 ^■ - 62 GSavSA

103 - 108 GGdaGV

107 - 112 GVqiCL

136 - 141 GIcvSS

159 - 164 GNdhST

183 - 188 GQegSV

198 - 203 GLccAR

217 - 222 GQvcTK

151P4E11

N-glycosylation site

1 - 4 NSSR

55 ^■ - 58 NASE

168 - 171 NASE

284 - 287 NASE cAMP- and cGMP-dependent protein kinase phosphorylation site

131 - 134 RRtT

135 - 138 RKgS

246 - 249 RRsS Protein kinase C phosphorylation site 2 - 4 SsR

45 - 47 TpR

81 ^■ - 83 TeR

95 ^■ - 97 TeR

133 - 135 TtR

134 - 136 TrK

144 - 146 SpK

195 - 197 TeR

209 - 211 TeR

245 - 247 SrR

248 - 250 SsK

251 - 253 ScR

254 - 256 SsR

262 - 264 ScK

312 - 314 TeR

326 - 328 TeR

406 - 408 TeR

420 - 422 TeR

Caseinkinase II phosphorylation site

3 - 6 SrpD

22 - 25. SitE

74 ■ - 77 SitE

155 - 158 SitE

188 - 191 SitE '

255 - 258 SrpD

272 - 275 SitE

305 - 308 SitE

362 - 365 TitE

366 - 369 StlD

399 - 402 SitE

497 - 500 SgπD

503 - 506 TgqE

N-myristoylation site

456 - 461 GAagAT 462 - 467 GVsaTL

468 - 473 GGaaGH

474 - 479 GSavSA

480 - 485 GGdaGV

486 - 491 GVqiCL

492 - 497 GIcvSS

498 - 503 GNdhST

504 - 509 GQegSV

510 - 515 GLccAR

516 - 521 GQvcTK

154P2A8

N-glycosylation site 4 - 7 NLTL 25 - 28 NRSD 33 - 36 NTTL 72 - 75 NKTS 251 - 254 NQSI Protein kinase C phosphorylation site 236 - 238 SsR

244 • - 2 24466 SSssRR

245 • - 2 24477 SSrrKK

253 ^■ - 225555 SSiiRR

322 • - 332244 SSrrRR

337 ^■ - 333399 SSiiRR

343 - 345 SvR Casein kinase II phosphorylation site 278 - 281 ShlD 333 - 336 TrsE Tyrosine kinase phosphorylation site 145 - 153 KpfgDsrmY 345 - 352 Rrs . EvriY N-myristoylation site

224 - 229 GCyiAI G-protein coupled receptors family 1 signature

127 - 143 TSIvfLgllSIDRYlkV G-protein coupled receptors family 1

59 - 314 LNGIiAVWIFFHIRN-KTSFIFYLKNIVVADLI TLTFPFRIVHDAGFGP YFKFILCRYT SVLFYANMYTSIVFLGLISIDRYLKWKPFGDS_røSITFTIWLSVCT/WIMAVLSLPNI

I -LTNGQPTEDNIHDCSKLK SPLGVK HTAVTyvNSCLFVAVLVILIGCYIAISRYI

HKSSRQF ISQSSRKRKHNQSIRVWAVFFTCFLPYHLCRIPFTFShldrlLDESA

QKILYYCKEITLFLSACNVCLDPI IY

156P1D4

N-glycosylation site

76 - 79 NVTQ

93 - 96 NHTL

Tyrosine sulfation site

41 - 55 awdtneeYl:

Protein kinase C phosphorylation site

30 - 32 SiR

57 - 59 SmR

78 - 80 TqR

Casein kinase II phosphorylation site

44 - 47 TneΞ 177 - 180 SevD

214 - 217 TedE N-myristoylation site

20 - 25 GAenAF

148 - 153 GVifCI

205 - 210 GGhiND

156P5C12

Protein kinase C phosphorylation site

12 - 14 SdR

32 - 34 TfR

103 - 105 SeR

131 - 133 TlR

221 - 223 SsK

Casein kinase II phosphorylation site

131 - 134 TlrE N-myristoylation site

48 - 53 GGplAL

120 - 125 GMvgAL

152 - 157 GlakAL

159P2B5 cAMP- and cGMP-dependent protein kinase phosphorylation site

168 - 171 RRhT N-myristoylation site

27 - 32 GNppTG

38 - 43 GSppGG

43 - 48 GGflGS

64 - 69 GQdvSR

122 - 127 GAtlSA

149 - 154 GIlgGV

158 - 163 GSraCT

161P2B7A

N-glycosylation site 17 - 20 NFTL 171 - 174 NSSI Tyrosine sulfation site

28 - 42 rlfdethYpdafmre Protein kinase C phosphorylation site 45 - 47 SqR 168 - 170 TsK Casein kinase II phosphorylation site 83 - 86 SqfE 173 - 176 SiaD Tyrosine kinase phosphorylation site

28 - 35 RlfdEth .Y

N-myristoylation site .

49 - 54 GLseAR

76 - 81- GVliGA

142 - 147 GLplAT

' Homeobox ' domain signature

44 - 67 LSqrLgLsearVQVWFqNrrakcR ¹ Homeobox ' domain

9 - 69 IKQRRSRTNFTLEQLNELERLFDETHYPDAFMREELSQRLGLSEARVQ FQNRRAKCRK OAR domain

172 - 185 SSIADLRLKAKKHA

179P3G7

N-glycosylation site

257 - 260 NTTG

306 - 309 NLTD

Protein kinase C phos iphorylation site

54 - 56 SkR

104 - 106 SvK

201 - 203 TpK

230 - 232 SeK

264 - 266 TaK

267 - 269 SgR

308 - 310 TdR

Casein kinase II phosphorylation site

11 - 14 SyaE

35 - 38 SgsD

54 - 57 SkrD

74 - 77 SqlD

78 - 81 SwgD

104 - 107 SvkE

122 - 125 SgpE

156 - 159 SalD

197 - 200 SfpE

210 - 213 SpnE

226 - 229 SpsE

228 - 231 SesE

230 - 233 SekE

240 - 243 SspD

245 - 248 SdnE

Tyrosine kinase phosphorylation site 121 - 129 KsgpEaalY N-myristoylation site

36 - 41 GSdfNC

42 - 47 GVmrGC

48 - 53 GLapSL

59 - 64 GSspSL

193 - 198 GGkvSF

260 - 265 GNwlTA

Amidation site

267 - 270 sGRK 'Homeobox' domain signature

301 - 324 ISktlnLtdrqVKIWFqNrrmklK 'Homeobox' domain

266 - 326 KSGRKKRCPYTKHQTLELEKEFLF YLTRERRLEISKTINLTDRQVKIWFQNRRMKLKK M

184P3C10B

N-glycosylation site

10 - 13 NATL

64 - 67 NTSM

184 - 187 NLTL

202 - 205 NASF

362 - 365 NLTC 367 - 370 NQTQ Protein kinase C phosphorylation site 33 - 35 TcK 186 - 188 TlK Casein kinase II phosphorylation site 69 - 72 ThpD 169 - 172 ThgD 259 - 262 TqnE 330 - 333 SsfD N-myristoylation site

308 - 313 GLkpAS 316 - 321 GirtSG Bipartite nuclear targeting sequence

122 - 138 RRellrrtwgrerkvrg TonB -dependent receptor proteins signature 1

1 - 32 mkylrhrrpnatlilaigaf till FSLLV sPP

184P3G10

N-glycosylation site

440 - 443 NLSS

678 - 681 NFTI Tyrosine sulfation site

336 - 350 gravqavYedmdark Protein kinase C phosphorylation site

63 - 65 SsR

105 - 107 SgK

188 - 190 StR

648 - 650 ThK

663 - 665 SrR Casein kinase II phosphorylation site

13 - 16 TkyD

46 - 49 SpgΞ

84 - 87 TflE

301 - 304 SeeE

373 - 376 SglE

422 - 425 SflE

540 - 543 SaeE

553 - 556 TdgD

570 - 573 SgaD 737 - 740 SaqE

N-myristoylation site

59 - 64 GLpfSS

104 - 109 GSgkST

413 - 418 GSclCL

493 - 498 GVilSE

571 - 576 GAdeA 581 - 586 GLnwAA 614 - 619 GSeqNR 725 - 730 GMadST 732 - 737 GSksSS

ATP/GTP-binding site motif A (P-loop)

101 - 108 GepgsGKS

Prenyl group binding site (CAAX box)

745 - 748 CVLq 185P2C9 var 1

N-glycosylation site

834 - 837 NKS 1000 - 1003 NLSD 1033 - 1036 NGSR 1129 - 1132 NRTS 1296 - 1299 NQTV Tyrosine sulfation site

822 - 836 IspddlkYieefnks cAMP- and cGMP-dependent protein kinase phosphorylation site

155 - 158 KKeS

Protein kinase C phosphorylation site

102 - 104 SaR

111 - 113 SgK

143 - 145 SpR

192 - 194 SgK

218 - 220 TlK

360 - 362 SIR

434 - 436 S1K

496 - 498 SdR

- 519 - 521 SIR

571 - 573 SdK

639 - 641 SeK

680 - 682 SpR

704 - 706 SIR

735 - 737 SdR

813 - 815 SsK

937 - 939 TsR

963 - 965 SpR

1011 - 1013 SvR

1069 - 1071 TpK

1085 - 1087 SIR

1108 - 1110 SpK

1113 - 1115 SpK

1150 - 1152 TtR

Casein kinase II phosphorylation site

32 - 35 SstE

53 - 56 SisE

55 - 58 SeiE

143 - 146 SprD

147 - 150 SdaE

158 - 161 SdgE

178 - 181 SdsE

235 - 238 TdtD

415 - 418 TlhE

420 - 423 T sD

467 - 470 TegE

603 - 606 SsaE

739 - 742 SasE

750 - 753 SldD

790 - 793 SmsE

813 - 816 SskE

814 - 817 SkeD

823 - 826 SpdD 890 - 893 TspE

943 - 946 SpgD

1150 - 1153 TtrE

1159 - 1162 TinD

1246 - 1249 TltE

1284 - 1287 SlgD

Tyrosine kinase phosphorylation site

208 - 215 KareDse . Y

N-myristoylation site

85 - 90 GAspGA

89 - 94 GAggGA

245 - 250 GLrgGA

275 - 280 GTinAK

383 - 388 GGqtCF

482 - 487 GVqgGH

485 - 490 GGhqAD

685 - 690 GSflCD

808 - 813 GLpsTS

1057 - 1062 GSgvTS

1184 - 1189 GLraGS

1203 - 1208 GQetGT

1221 - 1226 GSemCR

Amidation site

153 - 156 aGKK

Cell attachment sequence

354 - 356 RGD

185P2A9 var 2

N-glycosylation site

834 - 837 NKS 1000 - 1003 NLSD 1033 - 1036 NGSR Tyrosine sulfation site

822 - 836 IspddlkYieefnks cAMP- and cGMP-dependent protein kinase phosphorylation site

155 - 158 KKeS Protein kinase C phosphorylation site

102 - 104 SaR

111 - 113 SgK

143 - 145 SpR

192 - 194 SgK

218 - 220 T1K

360 - 362 SIR

434 - 436 SlK

496 - 498 SdR

519 - 521 SIR

571 - 573 SdK

639 - 641 SeK

680 - 682 SpR

704 - 706 SIR

735 - 737 SdR

813 - 815 SsK

937 - 939 TsR

963 - 965 SpR

1011 - - 1013 SvR 1069 - 1071 TpK

1085 - 1087 SIR

1108 - 1110 SpK

1113 - 1115 SpK

Casein kinase II phosphorylation site

32 - 35 SstE

53 - 56 SisE

55 - 58 SeiE

143 - 146 SprD

147 - 150 SdaB

158 - 161 SdgE

178 - 181 SdsE

235 - 238 TdtD

415 - 418 TlhE

420 - 423 T sD

467 - 470 TegE

603 - 606 SsaE

739 - 742 SasE

750 - 753 SldD

790 - 793 SmsE

813 - 816 SskE

814 - 817 SkeD

823 - 826 SpdD

890 - 893 TspE

943 - 946 SpgD

Tyrosine kinase phosphorylation site 208 - 215 KareDse . Y

N-myristoylation site

85 - 90 GAspGA 89 - 94 GAggGA

245 ^■ - 250 GLrgGA

275 - - 280 GTinAK

383 - - 3 38888 GGqtCF

482 • - 4 48877 GVqgGH

485 - 490 GGhqAD

685 - 690 GSflCD 808 - 813 GLpsTS ' 1057 - 1062 GSgvTS Amidation site. .

153 - 156 aGKK

185P2A9 var 3

N-glycosylation site

844 - 847 NKSW

1010 - 1013 NLSD

1043 - 1046 NGSR

1139 - 1142 NRTS

Tyrosine sulfation site

832 - 846 IspddlkYieefnks cAMP- and cGMP-dependent protein kinase phosphorylation site

155 - 158 KKeS Protein kinase C phosphorylation site

102 - 104 SaR

111 - 113 SgK

143 - 145 SpR 192 - 194 SgK

218 - 220 TiK

360 - 362 SIR

434 - 436 SIK

496 - 498 SdR

519 - 521 SIR

571 - 573 SdK

614 - 616 TvK

617 - 619 TiK

677 - 679 SeK

714 - 716 SIR

745 - 747 SdR

823 - 825 SsK

947 - 949 TsR

973 - 975 SpR

1021 - 1023 SvR

1079 - 1081 TpK

1095 - 1097 SIR

1118 - 1120 SpK

1123 - 1125 SpK

1160 - 1162 TtR

Caseinkinase II phosphorylation site

32 - 35 SstE

53 - 56 SisE

55 - 58 SeiE

143 - 146 SprD

147 - 150 SdaE

158 - 161 SdgE

178 - 181 SdsE

235 - 238 TdtD

415 - 418 TlhE

420 - 423 TwsD

467 - - 447700 T TeeggEE

641 - - 664444 S SssaaEE

749 - - 775522 S SaassEE

760 - ^• 776633 S SllddDD

800 - - 8 80033 SSmmssEE

823 ^■ - 8 82266 SSsskkEE

824 - ^■ 8 82277 SSkkeeDD

833 - - 883366 SSppddDD

900 ^■ - 990033 TTssppEE

953 - - 995566 SSppggDD

1160 - 1163 TtrE

1169 - 1172 TinD

1256 - 1 1225599 TTllttEE

1294 - 11229977 SSllggDD

Tyrosine kinase phosphorylation site

208 ^• - 215 KareDse.Y

N-myristoylation site

85 - 90 GAspGA

89 - 94 GAggGA

245 ^■ - 250 GLrgGA

275 ^■ - 280 GTinAK

383 - - 388 GGqtCF

482 ^■ - 487 GVqgGH

485 ^■ - 490 GGhqAD

818 ^■ - 823 GLpsTS 1067 - 1072 GSgvTS

1194 - 1199 GLraGS

1213 - 1218 GQetGT

1231 - 1236 GSemCR

Amidation site

153 - 156 aGKK

Cell attachment sequence

354 - 356 RGD

185P3C2

N-elvcosvlation site

8 - 11 NKSV

92 - 95 NGSL

Tyrosine sulfation site

328 - 342 qeqtdfaYdsdvtgc

Protein kinase C phosphorylation site

10 - 12 SvR

46 - 48 TpR

65 - 67 SgR

86 - 88 SsK

94 - 96 SIR

159 - 161 TtR

168 - 170 SpR

178 - 180 SrK

291 - 293 SfK

430 - 432 TgR

465 - 467 SIR

Casein kinase II phosphorylation site

94 - 97 SlrE

195 - ^■ 198 SayD

356 - ^• 359 SpgD

516 - ^■ 519 SeeD

524 - ^■ 527 ShlD

Tyrosine kinase phosphorylation site

477 - ^■ 485 KvagEryvY

N-myristoylation site

16 - 21 GAgaAL

32 - 37 GSraCS

312 - ^• 317 GGvnGH

449 - ^■ 454 GlqkNR

Ets-domain signature 1.

410 - - 418 LWQFLVaLL

Ets-domain signature 2

454 - ^• 469 RpaMnYDkLSRSLRyY

Ets-domain

408 - 488 j l_QLWQFLVALLDDPTNAHFIAWTGRGMEFKLIEPEEVARLWGIQ_aJRPAMNYDKLSRSLR -TYYEKGIMQKVAGERYVYKFV

186P1H9

N-glycosylation site

147 - 150 NVSN

188 - 191 NETS

392 - 395 NCST Protein kinase C phosphorylation site

79 - 81 SaR

87 - 89 TgK

107 - 109 TgK

132 - 134 TIK

316 - 318 TtR

Casein kinase II phosphorylation site

54 - 57 SpeE

107 - 110 TgkD

111 - 114 TmgD

132 - 135 TlkD

209 - 212 TelE

242 - 245 TlpE

316 - 319 TtrD

422 - 425 TceE

N-myristoylation site

95 - 100 GLagGK

103 - 108 GAgaTG

264 - 269 GTpfSY

320 - 325 GMweAF

332 - 337 GTgeNL

Leucine zipper pattern

165 - 186 LgelerqLlrkvaeLedekslL

Pentaxin family signature

310 - 317 HiCvTWtT

187P3F2 cAMP- and cGMP-dependent protein kinase phosphorylation site

408 - 411 KKrT

409 - 412 KRtS

462 - 465 KRmT Protein kinase C phosphorylation site 69 - 71 SvK

369 - 371 SfK

416 - 418 SvK Casein kinase II phosphorylation site

312 - 315 SdeD

318 - 321 TsdD

339 - 342 TqaD

395 - 398 TsiD

411 - 414 TsiE

432 - 435 SaqE

473 - 476 TpdD stoylati on site

28 - 33 GAggGG

30 - 35 GGggGG

31 - 36 GGggGG

32 - 37 GGggGG

33 - 38 GGggGG

34 - 39 GGggGS

35 - 40 GGggSG

36 - 41 GGgsGG

37 - 42 GGsgGG

38 - 43 GSggGG

40 - 45 GGggAG 41 - 46 GGgaGG 42 - 47 GGagGG

43 - 48 GAggGG

45 - 50 GGggGG

46 - 51 GGggGM

50 • - 55 G qpGS

61 - 66 GAyrGD

80 - 85 GAmaAS

124 - 129 GSavG

185 - 190 GGwgAA

188 - 193 GAaaAA

241 - 246 GGggGG

242 - 247 • GGggGA

243 - 248 GGggAG

244 - 249 GGgaGG

245 - 250 GGagGG

246 - 251 GAggGA

249 - 254 GGaqSL

258 - 263 GLvrGD

295 - 300 GGggGG

296 - 301 GGggGA

297 - 302 GGggAG

300 - 305 GAgpGL

344 - 349 GLalGT

348 - 353 GTlyGN

352 - 357 GNvfSQ

392 - 397 GSptSI

419 - 424 GAleSH

482 - 487 GTvsAD

494 - 499 GLqtSV

Amidation site

403 - 406 qGRK

Cell attachment sequence

64 - 66 RGD

261 - 263 RGD

'Homeobox' domain signature

439 - 462 LAdsLqLekevVRVWFcNrrqkeK

'Homeobox' domain

404 - • _64 GRKRIOCRTSIEVSVKGALESHFLKCPKPSAQEITNLADSLQLEPCEVVRVWFCNRRQKEKR

'POU' domain signature 1

332 - 344 RRIkLGFtQaDVG

'POU' domain s signature 2

356 - 369 SQTTICRFEaLqLS

192P2G7

Protein kinase C phosphorylation site 104 - 106 SpR 148 - 150 SIR 153 - 155 SyR Casein kinase II phosphorylation site 4 - 7 SeaE 11 - 14 TpgE 157 - 160 TfqE 174 - 177 SwfE ^' 205 - 208 TmvE 260 - 263 SmnE Tyrosine kinase phosphorylation site 164 - 172 RfmnDklgY

TABLE XXI: Properties of Gene/Protein of Figure 2

74P3B3 var. lA Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bp289-831 (includes stop)

Protein length 180aa

Transmembrane TM Pred xh.embnet.org/ no TM region

HMMTop .enzim.hu/hmmtop/ no TM, intracellular

Sosui .genome.ad.jp/SOSui/ no TM, soluble protein

TMHMM .cbs.dtu.dk/services/TMHMM no TM

Signal Peptide Signal P .cbs.dtu.dk/services/SignalP/ no

Pi pl/MW tool .expasy.ch/tools/ pI5.5

Molecular weight pl/MW tool .expasy.ch/tools/ 20.2kDa

Localization PSORT psort.nibb.ac.jp/ 46% peroxisome, 45% cytoplasmic PSORT II psort.nibb.ac.jp/ 39% nuclear, 22% mitochondrial,

22% cytoplasm

Motifs Pfam . sanger. ac. uk/Pfam/ retroviral GAG plO protein

Prints .biochem.ucl.ac.uk/ none

Blocks .blocks.fhcrc.org/ RNA polymerases K 14 to 18kD subunits

74P3B3 var.lB Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bp756-1442 (includes stop)

Protein length 228aa

Transmembrane TM Pred xh.embnet.org/ 1TM 189-208aa, N terminus region intracellular

HMMTop . enzim. hu/hmmtop/ no TM, extracellular

Sosui .genome.ad.jp/SOSui/ no TM, soluble protein

TMHMM xbs.dtu.dk/services/TMHMM no TM

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ no pi pl/MW tool .expasy.ch/tools/ pI7.0

Molecular weight pl/MW tool .expasy.ch/tools/ 25.8kDa

Localization PSORT psort.nibb.ac.jp/ 74% peroxisome, 24% lysome (lumen)

PSORT II psort.nibb.ac.jp/ 56.5%cytoplasm, 26% nuclear

Motifs Pfam .sanger.ac.uk/Pfam/ none

Prints .biochem.ucl.ac.uk/ none

Blocks .blocks.fhcrc.org/ Omptin serine protease family

83P4B8 var.l Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bp25-4011 (includes stop)

Protein length 1328aa

Transmembrane TM Pred xh.embnet.org/ 2 TM helices (452-474aa, 739- region

763 aa), N terminus intracelluar

HMMTop .enzim.hu/hmmtop/ no TM, N terminus extracellular 83P4B8 var. l Bioinformatic URL on World Wide Web Outcome Program

Sosui .genome.ad.jp/SOSui/ no TM, soluble protein

TMHMM . cbs.dtu.dk services/TMHMM no TM, N terminus extracellular

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ no pi pl/MW tool .expasy.ch/tools/ pI6.3

Molecular weight pl/MW tool .expasy.ch/tools/ 149.3kDa

Localization PSORT psort.nibb.ac.jp/ 96% nuclear, 30% peroxisome

PSORT II psort.nibb.ac.jp/ 52% cytoplasmic, 22% nuclear 13% mitochondrial

Motifs Pfam .sanger.ac.uk Pfam/ none

Prints .biochem.ucI.ac.uk/ none

Blocks .bIocks.fhcrc.org/ aspartate-semi aldehyde dehydro genase, dihydrodipicolinate reductase, hydroxymethylglut- aryl-coenzyme A synthase, sigma 54 factor family, ribosomal protein S16

109P lD4 var.l Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bp846-3911 (includes stop)

Protein length 1021aa

Transmembrane TM Pred xh.embnet.org/ 3TM helices (3-23aa, 756-776aa, region

816-834aa), N terminus intracellular

HMMTop . enzim.hu/hmmtop/ no TM, N terminus extracellular

Sosui .genome.ad.jp/SOSui/ 3TM helices (2-24aa, 756-778aa, 810-832aa), N terminus extracellular

TMHMM xbs.dtu.dk/services/TMHMM 1TM helix (813-835aa), N terminus extracellular

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ yes pi pl/MW tool .expasy.ch/tools/ pI4.8

Molecular weight pl/MW tool .expasy.ch/tools/ 112.7kDa

Localization PSORT psort.nibb.aC.jp/ 40% plasma membrane

PSORT II psort.nibb.ac.jp/ 67% endoplasmic reticulum

Motifs Pfam .sanger.ac. uk Pfam/ Cadherin domain

Prints .biochem.ucl.ac.uk/ Cadherin domain

Blocks .blocks.fhcrc.org/ Cadherin domain, ribosomal protein L10E, ribulose biphos- phate carboxylase (large chain), ornithine decarboxylase antizyme protein phosphatase 2C subfamily

151PlC7A var.l Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bplO3-903 (includes stop)

Protein length 266aa Transmembrane TM Pred xh.embnet.org/ 2TM helices (8-29, 52-71) 151PlC7A var.l Bioinformatic URL on World Wide Web Outcome Program region

N terminus intracellular

HMMTop .enzim.hu/hmmtop/ 1TM helix (12-36aa), N terminus intracellular

Sosui .genome.ad.jp/SOSui/ no TM, soluble protein

TMHMM xbs.dtu.dk/services/TMHMM no TM

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ yes

Pi pl/MW tool .expasy.ch/tools/ pI8.8

Molecular weight pl/MW tool .expasy.ch/tools/ 28.7kDa

Localization PSORT psort.nibb.ac.jp/ 51% outside

PSORT II psort.nibb.ac.jp/ 52% nuclear, 17% cytoplasmic,

17% cytoskeletal

Motifs Pfam .sanger.ac.uk/Pfam/ none

Prints .biochem.ucl.ac.uk/ none

Blocks .blocks.fhcrc.org/ fungal transcriptional regulatory protein N terminus, transketolase granulin, chitin binding domain,

WAP-type four disulphide core, serine protease inhibitor (squash family, adenosine and AMP deaminase

151P4E11 var.l Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bp3-374 (includes stop)

Protein length 123aa Transmembrane TM Pred xh.embnet.org 1 TM helix (6-24aa) region

N terminus intracellular

HMMTop .enzim.hu/hmmtop/ 1TM helix (6-26aa), N terminus extracellular

Sosui .genome.ad.jp/SOSui/ 1TM helix 3-24aa), TMHMM xbs.dtu.dk/services/TMHMM 1TM helix (7-26aa) N terminus intracellular

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ yes

Pi pl/MW tool .expasy.ch/tools/ pI6.8

Molecular weight pl/MW tool .expasy.ch/tools/ 13.3kDa

Localization PSORT psort.nibb.ac.jp/ 82% outside

PSORT II psort.nibb.ac.jp/ 56% extracellular including cell wall, 22% nuclear

Motifs Pfam .sanger.ac.uk/Pfam/ galanin

Prints .biochem.ucI.ac.uk/ galanin signature, IL-8 signature Blocks .blocks.fhcrc.org/ galanin family, methyl malonyl- CoA mutase

154P2A8 var.l Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bp250-1326 (includes stop) Protein length 358aa 154P2A8 var. l Bioinformatic URL on World Wide Web Outcome Program

Transmembrane TM Pred xh.embnet.org/ 7TM helices (47-68, 79-99, 119- region

137, 160-180, 206-229, 253-271,

298-317aa)

N terminus extracellular

HMMTop .enzim.hu/hmmtop/ 7TM helices (43-67, 74-92, 119-

137, 160-179, 206-230, 253-271,

298-317aa)

N terminus extracellular

Sosui .genome.ad.jp/SOSui/ 6TM helices (43-65, 76-98, 127-

149, 157-179, 210-232, 296-

318aa)

TMHMM xbs.dtu.dk/services/TMHMM 7TM helices (47-69, 79-98, 118-

137, 157-179, 207-229, 256-278,

298-317aa)

N terminus extracellular

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ no pi pl/MW tool .expasy.ch tools/ pI9.55

Molecular weight pl/MW tool .expasy.ch/tools/ 41.4kDa Localization PSORT psort.nibb.ac.jp/ 60% plasma membrane, 40%

Golgi, 30% endoplasmic reticulum 30% peroxisome

PSORT II psort.nibb.ac.jp/ 56.5% plasma membrane, 13% vacuolar, 13% endoplasmic reticulum

Motifs Pfam .sanger.ac.uk/Pfam/ 7TM receptor (rhodopsin family)

Prints .biochem.ucl.ac.uk/ P2Y purinoceptor signature, rhodopsin like GPCR super- family signature

Blocks .blocks._hcrc.org/ rhodopsin like GPCR super- family, proton/sugar symporter (LacY family), amino acid perm- ease, xanthine/uracil permease family

156PID4 var.l Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bp24-692 (includes stop)

Protein length 222aa Transmembrane TM Pred xh.embnet.org/ 2TM helices (1-17, 142-167aa) region

N terminus extracellular

HMMTop .enzim.hu/hmmtop/ 1TM helix (140-164aa), N terminus extracellular

Sosui .genome.ad.jp/SOSui/ 2TM (l-23, 142-164aa) TMHMM xbs.dtu.dk/servjces/TMHMM 1TM helix (142-164aa) N terminus extracellular

Signal Peptide Signal P .cbs.dtu.dk/services/SignalP/ yes pi pl/MW tool .expasy.ch/tools/ pI5.4 156P1D4 var.l Bioinformatic URL on World Wide Web Outcome Program

Molecular weight pl/MW tool .expasyxh/tools/ 25.2kDa

Localization PSORT psort.nibb.ac.jp/ 46% plasma membrane

PSORT II psort.nibb.ac.jp/ 44% endoplasmic reticulum, 33% Golgi, 22% plasma membrane

Motifs Pfam .sanger.ac.uk/Pfam/ cytochrome B6-F complex sub- unit 5

Prints .biochem.ucl.ac.uk/ bacterial chemotaxis sensory transducer signature

Blocks .blocks.fhcrc.org/ nucleoside diphosphate kinase, integrins alpha chain, large conductance mechanosensitive channel mscL

156P5C12 var.1 Bioinformatic URL on World Wide Web Outcome

Program

ORF ORF finder bp 178-861 (includes stop)

Protein length 227aa Transmembrane TM Pred xh.embnet.org 3TM helices (60-78, 173-193, region

196-216aa) N terminus extracellular

HMMTop . enzim. hu/hmmtop/ 1TM helix (44-68aa),

N terminus intracellular

Sosui .genome.ad.jp/SOSui/ 1 TM helix (57-78aa)

TMHMM xbs.dtu.dk/services/TMHMM 1TM helix (43-65aa)

N terminus intracellular

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ yes pi pl/MW tool .expasy.ch/tools/ pI9.1

Molecular weight pl MW tool .expasy.ch tools 25.6kDa

Localization PSORT psort.nibb.ac.jp/ 60% plasma membrane, 40%

Golgi, 30% endoplasmic reticulum (membrane)

PSORT II psort.nibb.ac.jp/ 78% endoplasmic reticulum

Motifs Pfam . sanger. ac. uk/Pfam/ acetyltransferase (GNAT) family

Prints .biochem.ucl.ac.uk/ none

Blocks .blocks.fhcrc.org/ glycosylhydrolases family 16, elongation factor P (EF-P)

159P2B5 var.l Bioinformatic URL on World Wide Web Outcome

Program

ORF ORF finder bpl517-2191 (includes stop)

Protein length 224aa

Transmembrane TM Pred xh.embnet.org 2TM helices (90-109, 140-159aa) region

N terminus extracellular

HMMTop .enzim.hu/hmmtop/ no TM, N terminus intracellular

Sosui .genome.ad.jp/SOSui/ soluble protein

TMHMM xbs.dtu.dk/services/TMHMM 1 TM helix (140-159aa),

N terminus extracellular 159P2B5 var.l Bioinformatic URL on World Wide Web Outcome Program

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ no pi pl/MW tool .expasy.ch/tools/ pll l.6

Molecular weight pl/MW tool .expasyxh/tools/ 23.7kDa

Localization PSORT psort.nibb.ac.jp/ 70% plasma membrane, 51.5% peroxisome, 20% endoplasmic reticulum

PSORT II psort.nibb.ac.jp/ 70% nuclear, 13% plasma membrane

Motifs Pfam .sanger. ac. uk/Pfam/ none

Prints .biochem. ucl.ac. uk none

Blocks .blocks.fhcrc.org prion protein, ribosomal protein L10E, aminotransferase class- V

161P2B7 var.l Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bpl98-770 (includes stop)

Protein length 190aa

Transmembrane TM Pred xh.embnet.org/ 1TM helix (145-165aa) region

N terminus intracellular

HMMTop .enzim.hu/hmmtop/ 1TM helix (141-165aa), N terminus extracellular

Sosui .genome.ad.jp/SOSui no TM, soluble protein

TMHMM xbs.dtu.dk/services/TMHMM no TM

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ no pi pl/MW tool .expasy.ch/tools/ pI9.55

Molecular weight pl/MW tool .expasy.ch/tools/ 21kDa

Localization PSORT psort.nibb.ac.jp/ 65% cytoplasmic, 26% lysosome (lumen)

PSORT II psort.nibb.ac.jp/ 43.5% nuclear, 30% cytoplasmic

Motifs Pfam . sanger. ac. uk/Pfam/ Homeobox domain

Prints .biochemfucl.ac.uk/ lambda-repressor HTH signature, POU domain signature

Blocks .blocks.fhcrc.org/ Homeobox antennapedia-type protein, Homeobox domain, POU domain, involucrin, ribosomal protein LOe

179P3G7 var.l Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bp72-l 100 (includes stop)

Protein length 342aa Transmembrane TM Pred xh.embnet.org/ no TM region

HMMTop .enzim.hu/hmmtop/ no TM, N terminus intracellular

Sosui .genome.ad.jp/SOSui/ no TM, soluble protein

TMHMM xbs.dtu.dk/services/TMHMM no TM

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ no p pl/MW tool .expasy.ch/tools/ pI8.3 179P3G7 var.1 Bioinformatic URL on World Wid Outcome Program

Molecular wei ght pl/MW tool .expasy.ch/tools/ 38kDa

Localization PSORT psort.nibb.ac.jp/ 30% nuclear

PSORT II psort.nibb.ac.jp/ 83% nuclear, 17% mitochondrial

Motifs Pfam .sanger. ac. uk/Pfam/ Homeobox domain Homeobox antennapedia-type protein, Homeobox domain,

Prints .biochem.ucl.ac.uk/ lambda-repressor HTH signature, POU domain signature

Blocks .blocks.fhcrc.org/ Homeobox antennapedia-type protein, Homeobox domain, POU domain, adenylate cyclases class 1

184P3C10 var.l Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bp 118-1236 (includes stop)

Protein length 372aa

Transmembrane TM Pred xh.embnet.org/ 2TM helices (13-29, 264-282aa) region

N terminus intracellular

HMMTop .enzim.hu/hmmtop/ 1TM helix (1 l-29aa), N terminus intracellular

Sosui .genome.ad.jp/SOSui/ 1TM helix (12-32aa) TMHMM xbs.dtu.dk/services/TMHMM 1TM helix (13-30aa)

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ ^• yes pi pl MW tool .expasy.ch/tools/ pI8.9

Molecular weight pl MW tool .expasy.ch/tools/ 42.5kBa Localization PSORT psort.nibb.ac.jp/ 86.5% lysosyme (lumen), 82% outside, 18% nuclear

PSORT II psort.nibb.ac.jp/ 67% extracellular including cell wall

Motifs Pfam .sanger.ac.uk/Pfam/ . galactosyltransferase Prints .biochem.ucl.ac.uk/ none

Blocks .blocks.fhcrc.org/ CTF/NF-1 family, tyrosinase

184P3G10 var.l Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bp 14-2260 (includes stop)

Protein length 748aa

Transmembrane TM Pred xh.embnet.org/ 1TM helix (351 -371 aa) region

N terminus intracellular

HMMTop . enzim.hu/hmmtop/ no TM, N terminus intracellular

Sosui .genome.ad.jp/SOSui/ soluble protein

TMHMM xbs.dtu.dk services/TMHMM no TM, N terminus extracellular

Signal Peptide Signal P xbs.dtu.dk services/SignalP/ no pi pl MW tool .expasy.ch tools/ pI5.0

Molecular weight pl/MW tool .expasy.ch/tools/ 82.8kDa

Localization PSORT psort.nibb.ac.jp/ 65% plasma membrane, 45% 184P3G10 var.l Bioinformatic URL on World Wide Web Outcome Program cytoplasmic, 30%peroxisome

PSORT II psort.nibb.ac.jp/ 22%o nuclear, 22% plasma membrane, 17% mitochondrial, 13% vesicles of secretory system

Motifs Pfam .sanger.ac.uk/Pfam/ luminal portion of cytochrome b559, alpha (gene psbE) subunit

Prints . biochem. uci.ac.uk/ none

Blocks .blocks.fhcrc.org/ bacterial type II secretion system protein E, ribulose biphosphate carboxylase (large chain), unchar- acterized protein family UPF 0038, ATP-dependent helicase (DEAH-box subfamily), adenyl- late kinase, Tub family, GTP- binding elongation factor, AAA protein, legume lectin alpha domain, thymidylate kinase, Sigma-54 transcription factor family, RNA 3'-terminal phosphatase cyclase

185P2C9 var.l Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bp 140-4063 (includes stop)

Protein length 1307aa

Transmembrane TM Pred xh.embnet.org/ no TM region

HMMTop .enzim.hu hmmtop/ no TM, N terminus intracellular

Sosui .genome.ad.jp/SOSui/ soluble protein

TMHMM xbs.dtu.dk/services/TMHMM no TM, N terminus extracellular

Signal Peptide Signal P xbs.dtu.dk services/SignalP/ no pi pl/MW tool .expasy.ch tools/ pI5.7

Molecular weight pl/MW tool .expasy.ch/tools/ 145.4kDa

Localization PSORT psort.nibb.ac.jp/ 45% cytoplasmic, 30% peroxisome

PSORT II psort.nibb.ac.jp/ 70% nuclear, 22% cytoplasmic

Motifs Pfam .sanger.ac.uk/Pfam/ GLTT repeat (6 copies)

. Prints .biochem.ucl.ac.uk/ cadherin signature

Blocks .bIocks.fhcrc.org/ transketolase, glypican, sigma 54 factor family, receptor tyrosine kinase class V, DNA topoisomer- ase II family, 3'5'-cyclic nucleotide phosphodiesterase

185P2C9 var.2 Bioinformatic URL on World Wide Web Outcome

Program

ORF ORF finder bp 140-3568 (includes stop)

Protein length 1142aa

Transmembrane TM Pred xh.embnet.org/ no TM region 185P2C9 var.2 Bioinformatic URL on World Wide Web Outcome Program

HMMTop .enzim.hu/hmmtop/ no TM, N terminus intracellular

Sosui .genome.ad.jp/SOSui/ soluble protein

TMHMM xbs.dtu.dk/services/TMHMM no TM, N terminus extracellular

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ no pi pl MW tool .expasy.ch/tools/ pI5.7

Molecular weight pl/MW tool .expasy.ch/tools/ 127.9kDa

Localization PSORT psort.nibb.ac.jp/ 45% cytoplasmic, 30% peroxisome

PSORT II psort.nibb.ac.jp/ 70%> nuclear, 22% cytoplasmic

Motifs Pfam .sanger.ac.uk/Pfam/ GLTT repeat (6 copies)

Prints .biochem.ucl.ac.uk/ cadherin signature

Blocks .blocks.flicrc.org/ transketolase, glypican, sigma 54 factor family, DNA topoisomer- ase II family, 3'5'-cyclic nucleotide phosphodiesterase

185P2C9 var.3 Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bp 140-4080 (includes stop)

Protein length 1313aa

Transmembrane TM Pred xh.embnet.org/ no TM region

HMMTop .enzim.hu/hmmtop/ no TM, N terminus intracellular

Sosui .genome.ad.jp/SOSui/ soluble protein

TMHMM xbs.dtu.dk/services/TMHMM no TM, N terminus extracellular

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ no

Pi pl/MW tool .expasy.ch/tools/ pI5.6

Molecular weight pl/MW tool .expasy.ch/tools/ 145.9kDa

Localization PSORT psort.nibb.ac.jp/ 45% cytoplasmic, 30% peroxisome

PSORT II psort.nibb.ac.jp/ 70% nuclear, 22% cytoplasmic

Motifs Pfam .sanger.ac.uk/Pfam/ GLTT repeat (6 copies)

Prints .biochem.ucl.ac.uk/ cadherin signature

Blocks .blocks.fhcrc.org/ transketolase, glypican, sigma 54 factor family, receptor tyrosine kinase class V, DNA topoisomer- ase II family, 3'5'-cyclic nucleotide phosphodiesterase

185P3C2 var.1 Bioinformatic URL on World Wide Web Outcome

Program

ORF ORF finder bp3-1658 (includes stop)

Protein length 551aa

Transmembrane TM Pred xh.embnet.org/ no TM region

HMMTop •enzim.hu/hmmtop/ no TM, N terminus intracellular

Sosui •genome.ad.jp/SOSui/ soluble protein

TMHMM xbs.dtu.dk/services/TMHMM no TM, N terminus extracellular

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ no 185P3C2 var. l Bioinformatic URL on World Wid Outcome Program pi pl MW tool .expasy.ch/tools/ pI6.0

Molecular weight pl/MW tool .expasy.ch/tools/ 60.6kDa

Localization PSORT psort.nibb.ac.jp/ 48%peroxisome, 48% cytoplasmic, 18% lysosome(lumen)

PSORT II psort.nibb.ac.jp/ 52% mitochondrial, 39% nuclear

Motifs Pfam .sanger.ac.uk/Pfam/ Ets-domain

Prints .biochem.ucl.ac.uk/ Ets-domain signature

Blocks .blocks._hcrc.org/ Ets-domain, uncharacterized protein family UPF0035, bacterial regulatory proteins MerR family, cereal trypsin alpha amylase inhibitors family, purine and other phosphorylases family 2

186P 1 H9 var.1 Bioinformatic URL on World Wide Web Outcome

Program

ORF ORF finder bpl 70-1462 (includes stop)

Protein length 430aa

Transmembrane TM Pred xh.embnet.org/ 2TM helices (1-17, 246-265aa) region

N terminus extracellular

HMMTop .enzim.hu/hmmtop/ 1TM helix (246-265aa) N terminus intracellular

Sosui .genome.ad.jp/SOSui/ 1TM helix (1-16aa)

TMHMM xbs.dtu.dk/services/TMHMM no TM, N terminus extracellular

Signal Peptide Signal P .cbs.dtu.dk/services/SignalP/ no p pl/MW tool .expasy.ch/tools/ pI5.4

Molecular weight pl MW tool .expasy.ch/tools/ 46.8kDa

Localization PSORT psort.nibb.ac.jp/ 68% outside, 48% lysosome (lumen)

PSORT II psort.nibb.ac.jp/ 56.5%cytopIasmic, 22% nuclear 17% mitochondrial

Motifs Pfam .sanger.ac.uk/Pfam/ pentaxin family

Prints .biochem.ucl.ac.uk pentaxin signature

Blocks .blocks.fhcrc.org/ pentaxin family, interleukin-4 and- 13, amyloidogenic glycoprotein (amyloid A4)

187P3F2 var.1 Bioinformatic URL on World Wide Web Outcome

Program

ORF ORF finder bp60-1562 (includes stop)

Protein length 500aa

Transmembrane TM Pred xh.embnet.org/ 4TM helices (26-51, 100-122, region

185-204, 336-355aa) N terminus extracellular

HMMTop xnzim.hu/hmmtop/ 1TM helix (336-355aa) N terminus intracellular 187P3F2 var. l Bioinformatic URL on World Wide Web Outcome Program

Sosui •genome.ad.jp/SOSui/ soluble protein

TMHMM xbs.dni.dk/services/TMHMM no TM, N terminus extracellular

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ no

Pi pl/MW tool .expasy.ch/tools/ pI7.5

Molecular weight pl/MW tool .expasy.ch/tools/ 50.4kDa

Localization PSORT psort.nibb.ac.jp/ 70% nuclear, 54% peroxisome, 33% lysosome(lumen)

PSORT II psort.nibb.ac.jp/ 100% nuclear

Motifs Pfam .sanger.ac.uk/Pfam/ POU domain - N terminal to homeobox domain, homeobox domain

Prints .biochem.ucl.ac.uk/ POU domain

Blocks .blocks.fhcrc.org/ POU domain, homeobox domain, homeobox antennapedia-type protein, synapsin, prion protein, glycosyl hydrolases family 45, glycoprotease M22 metallo- • protease family, histone H4, ribulose biphosphate carboxylase large chain, galacto- kinase, 'paired box' domain, hemolysin type calcium binding region, complement Clq protein, iron containing alcohol dehydrogenases, GHMP kinases putative ATP binding domain, phospho- glucose isomerase, pyridoxal- dependent decarboxylase family, acyl-coA dehydrogenase, ribosomal protein L15

MISSING AT THE TIME OF PUBLICATION

192P2G7 var.l Bioinformatic URL on World Wide Web Outcome Program

ORF ORF finder bp 84-938 (includes stop)

Protein length 284aa

Transmembrane T Pred xh.embnet.org/ no TM region

HMMTop . enzim. hu/hmmtop/ no TM, N terminus extracellular

Sosui .genome.ad.jp/SOSui/ soluble protein

TMHMM xbs.dtu.dk/services/TMHMM no TM, N terminus extracellular

Signal Peptide Signal P xbs.dtu.dk/services/SignalP/ no pi pl/MW tool .expasy.ch/tools/ pI5.4

Molecular weight pl/MW tool .expasy.ch/tools/ 33.1kDa

Localization PSORT psort.nibb.ac.jp/ 66% peroxisome, 10% mitochondrial space, 10% lysosome(lumen)

PSORT II psort.nibb.ac.jp/ 65% cytoplasmic, 17% nuclear

Motifs Pfam .sanger.ac.uk/Pfam/ sulfotransferase protein

Prints .biochem.ucl.ac.uk/ none

Blocks .blocks.fhcrc.org/ Tub family, ribulose biphosphate carboxylase(large chain), serum albumin family, GPCR family 3

TABLE XXII: Chromosomal localization of Genes of Figure 2.

htr ://wΛVΛV.brni-heidelberg.corn/sy_peithi

TABLE XXIII - 74P3B3 Clone B

(Frame 1) HLA-DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

11 Y L H F I K L L L K R A G I K 29

15 I K L L L K R A G I K A S T E ^• 27

78 w S N A I V K A A S E P F Q 27

69 E G K F I P L T A W S N W A I 26

134 L I S F E E H V G P S A A P K 26

32 I T L F P T V E Q Y c P W F P 25

42 c P W F P E H G T M D F K D w 25

80 N w A I V K A A s E P F Q S E 25

101 A E R I S A E E G G D A A Ξ G 25

138 E E H V G P S A A P K I E K P 25

155 P R C L K Q R R A L R s S R L 25

158 L K Q R R A L R S S R L L I G 24

28 T E N L I T L F P T V E Q Y C 23

25 K A S T E N L I T L F P T V E 22

35 F P T V E Q Y c P W F P E H G 22

50 T M D F K D W E Q V G I A L K 22

52 D F K D W E Q V G I A L K Q V 22

135 I S F E E H V G P s A A P K I 21

132 D Ξ L I S F E E H V G P s A A 20

12 L H F I K L L L K R A G I K A 19

13 H F I K L L L K R A G I K A S 19

16 K L L L K R A G I K A s T E N 19

41 Y C P W F P E H G T M D F K D 19

62 A L K Q V C K E G K F I P L T 19

63 L K Q V C K E G K F I P L T A 19

75 L T A w S N W A I V K A A s E 19

95 N E A Y P P A E R I s A E E G 19

152 P Y M P R C L K Q R R A L R S 19

164 L R S s R L L I G I I R S G R 19

8 H S A Y L H F I K L L L K R A 18

88 S E P F Q S E N E A Y P P A E 18

144 S A A P K I E K P Y M P R C L 18

6 s K H s A Y- L H F I K L L L K 17

14 F I K L L L K R A G I ■K A s T 17

2.1 R A G I K A S T E N L I T L F 17

48 H G T M D F K D W E Q V G I A 17

53 F K D W E Q V G I A L K Q V C 17

60 G I A L K Q V C K E G K F I P 17

68 K Ξ G K F I P L T A W s N w A 17

72 F I P L T S N W A I V K A 17

89 E P F Q S E N E A Y P P A E R 17

102 E R I S A E E G G D A A E G G 17

108 E G G D A A E G G E D S E E D 17

123 F E E N T D K P G D E L I s F 17

143 P S A A P K I E K P Y M P R C 17

20 K R A G I K A S T E N L I T L 16

29 E N L I T L F P T V E Q Y c P 16

38 V E Q Y C P W F P E H G T D 16

55 ^' D W E Q V G I A L K Q V c K E 16

66 V C K E G K F I P L T A w s N 16

79 S N W A I V K A A S E P F Q S 16

93 S E N E A Y P P A E R I S A E 16

147 P K I E K P Y M P R C L K Q R 16

159 K Q R R A L R S S R L L I G I 16 TABLE XXIII - 74P3B3 Clone B

(Frame 1) HLA-DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

163 A L R S S R L L I G I I R S G 16

165 R S S R L I G I I R s G R L 16

1 M G Q S K S K H s A Y L H F I 15

9 S A Y L H F I K L L L K R A G 15

34 L F P T V E Q Y C P W F P E H 15

82 A I V K A A S E P F Q S E N Ξ 15

86 A A S Ξ P F Q S E N E A Y P P 15

119 S E E D F E E N T D K P G D E 15

18 L L K R A G I K A S T E N L I 14

77 A s N W A I V K A A S E P F 14

98 Y P P A E R I S A E E G G D A 14

128 D K P G D Ξ L I S F E E H V G 14

136 S F E E H V G P S A A P K I E 14

TABLE XXIV - 74P3B3, clone B

(Frame 1) HLA-DRB1*0301

(DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

112 A A E G G E D S E E D F E Ξ N 26 12 L H F I K L L L K R A G I K A 20 21 R A G I K A S T Ξ N L I T L F 19 72 F I P L T A W S N W A I V K A 19 80 N W A I V K A A S E P F Q S E 19 126 N T D K P G D E L I S F E E H 19 131 G D E L I S F E E H V G P S A 19 8 H S A Y L H F I K L L L K R A 18

31 L I T L F P T V E Q Y C P W F 18

42 C P W F P E H G T M D F K D W 18

46 P E H G T.M D F K D W E Q V G 18

56 W E Q V G I A L K Q V C K E G 18

81 W A I V K A A S Ξ P F Q s Ξ N 18

116 G E D S E E D F E E N T D K P ^' 18

151 K P Y M P R C L K Q R R A L R 18

49 G T M D F K D W E Q V G I A L 17

50 T M D F K D W E Q V G I A L K 17

60 G I A L K Q V C K E G K F I P 17

63 L K Q V C K E G K F I P L T A 17

61 I A L K Q V C K E G K F I P L 16

87 A S E P F Q S E N E A Y P P A 16

88 S E P F Q S E N Ξ A Y P P A E 16

153 Y M P R C L K Q R R A L R S S 16

32 I T L F P T V E Q Y C P W F P 15

142 G P S A A P K I E K P Y M P R 15

29 E N L I T L F P T V E Q Y C P 14

9 S A Y L H F I K L L L K R A G 13

14 F I K L L L K R A G I K A S T 13

15 I K L L L K R A G I K A S T E 13

16 K L L L K R A G I K A S T E N 13

161 R R A L R S S R L L I G I I R 13

24 I K. A S T E N L I T L F P T V 12

58 Q V G I A'L K Q V C K E G K F 12

70 G K F I P L T A w S N W A I V 12

132 D E L I S F E E H V G P S A A 12

138 E E H V G P S A A P K I E K P 12

155 P R C L K Q R R A L R S S R L 12 TABLE XXIV - 74P3B3, clone B

(Frame 1-) HLA-DRB1*0301

(DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num. 166 S S R L L I G I I R S G R L Q 12

TABLE XXV - 74P3B3, clone B

(frame 1) HLA-DRB1*0401

(DR4D 4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

95 N E A Y P P A E R I S A E E G 28

29 E N L I T L F P T V Ξ Q Y C P 26

8 H S A Y L H F I K L L L K R A 22

11 Y L H F I K L L L K R A G I K 22 32 I T L F P T V Ξ Q Y C P W F P 22 50 T M D F K D W E Q V G I A L K 22 53 F K D W E Q V G I A L K Q V c 22 69 E G K F I P L T A W S N W A I 22 9 S Y L H F I K L L L K R A G 20

16 K L L L K R A G I K A S T E N 20

60 G I A L K Q V C K E G K F I P 20

72 F I P L T A W s N W A I V K A 20

80 N W A I V K A A S E P F Q s E 20

81 W A I V K A A S E P F Q S E N 20

101 A E R I s A E E G G D A A E G 20

132 D E L I s F E E H V G P S A A 20

13 H F I K L L L K R A G I K A S 18

20 K R A G I K A S T E N L I T L 18

57 E Q V G I A L K Q V C K E G K 18

71 K F I P L T A W S N W A I V K 18

118 D S E E D F E E N T D K P G D 18

152 P Y M P R C L K Q R R A L R S 18

158 L K Q R R A L R S S R L L I G 18

41 Y C P W F P E H G T M D F K D 16

42 C P F P E H G T M D F K D W 16

75 L T A W S N W A I V K A A S E 16

^'78 W S N W A I V K A A S E P F Q 16

88 S E F Q S E N E A Y P P A E 16

134 L I S F E E H V G P S A A P K 16

12 L H F I K L L L K R A G I K A 14

21 R A G I K A S T E N L I T L F 14

28 T E N L I T L F P T V E Q Y C 14

56 W E Q V G I A L K Q V C K Ξ G 14

63 L K Q V C K E G K F I P L T A 14

131 G D E L I S F E E H V G P S A 14

138 E E H V G P S A A P K I E K P 14

TABLE XXVI - 74P3B3, clone B

(frame 1) HLA-DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

12 L H F I K L L L K R A G I K A 27 11 Y L H F I K L L L K R A G I K 24 95 N E A Y P P A E R I S A E E G 24 32 I T L F P T V E Q Y C P W F P 22 77 A W S N W A I V K A A S E P F 22 63 L K Q V C K E G K F I P L T A 21 60 G I A L K Q V C K E G K F I P 20 132 D E L I S F E E H V G P S A A 20 TABLE XXVI - 74P3B3 , clone B

( frame 1) HLA-DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

8 H S A Y L H F I K L L L K R A 19

134 L I S F E E H V G P S A A P K 18

53 F K D W E Q V G I A L K Q V C 17

75 L T A W S N W A I V K A A S E 17

38 V E Q Y C P W F P E H G T D 16

69 E G K F I P L T A W S N W A I 16

78 W S N W A I V K A A S E P F Q - 16

88 S E P F Q S E N E A Y P P A E 16

120 E E D F E E N T D K P G D E L 16

151 K P Y M P R C L K Q R R A L R 16

56 E Q V G I A L K Q V C K E G 15

148 K I E K P Y M P R C L K Q R R 15

152 P Y M P R C L K Q R R A L R S 15

13 H F' I K L L L K R A G I K A S 14

143 P S A A P K I E K P Y M P R C ^' 14

155 P R C L K Q R R A L R S S R L 14

165 R S S R L L I G I I R S G R L 14

9 S A Y L H F I K L L L K R A G 13

15 I K L L L K R A G I K A S T Ξ 13

28 T E N L I T L F P T V E Q Y C 13

131 G D E L I s F E E H V G P S A 13

161 R R A L R s S R L L I G I I R 13

166 S S R L L I G I I R S G R L Q 13 http://www.b_in-heide_berg.corn/syfpeithi

TABLE XXIII - 74P3B3, clone B

(frame 3) HLA-DRB1*0101 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score

Num.

42 V S R M N E L W P Q E P Q A H 31

75 P L Q F I I R Q A R L A G D L 30

114 P F S F K L L K D L K A A V G 29

152 P C D W E I L T K V T L S P S 29

194 A I T F E Q L L G I G G Q W G 28

134 S P F I R S L L Q s V A Q N K 26

213 H Q D F E M M P L N K F A I A 26

117 F K L L K D L K A A V G Q Y G 25

155 W E I L T K V T L s P S Q F L 25

168 F L Q F K T W W T D E A Q N Q 25

58 V A P V Q H K A A L P S N V N 24

83 A R L A G D L D A W Q F A V V 24

158 L T K V T L S P S Q F L Q F K 24

24 P ^•I K Q C S L E P w R S E S Q 23

55 A H G V A P V Q H K A A L P S 23

93 Q F A V V L Q P P R Q Q G G A 23

108 H Q A V W E P F s F K L L K D 23

123 L K A A V G Q Y G P N S P F I 23

124 K A A V G Q Y G P N S P F I R 23

138 R S L L Q S V A Q N K L L T P 23 •

157 I L T K V T L S P S Q F L Q F 23

197 F E Q L L G I G G Q W G T V N 23

200 L L G I G G Q W G T V N N H Q 23

5 K K G L E E Q s A P H W D H P 22

21 W P P P I K Q c s L E P W R S 22

39 I C P V S R N E L P Q E P 22 TABLE XXIII - 74P3B3, clone B

(frame 3) HLA-DRB1*0101 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 Seq. ID score Num.

87 G D L D A W Q F A V V L Q P P 22

154 D W E I L T K V T L S P S Q F 22

45 M N E L W P Q E P Q A H G V A 21

30 L E P W R s E S Q I C P V s R 20

112 W E P F S F K L L D L K A A 20

180 Q N Q D R K N R A A N P A I A 20

192 A I A I T F E Q L L G I G G Q 20

13 A P H W D H P E W P P P I K Q 19

89 L D A W Q F A V V L Q P P R Q 19

109 Q A V W E P F S F K L L K D L 19

171 F K T W W T D E A Q N Q D R K 19

190 N P A I A I T F E Q L L G I G 19

2 F K T K K G L E E Q S A P H W 18

18 H P E w P P P I K Q C S L E P 18

47 E L W P Q E P Q A H G V A P V 18

116 S F K L L K D L K A A V G Q Y 18

133 N S P F I R S L L Q S V A Q N 18

57 G V A P V Q H K A A L P S N V 17

68 P S N V N E S P L Q F I I R Q 17

74 S P L Q F I I R Q A R L A G D 17

98 L Q P P R Q Q G G A H Q A V W 17

99 Q P P R Q Q G G A H Q A V W E 17

120 L K D L K A A V G Q Y G P N S 17

121 K D L K A A V G Q Y G P N S P 17

143 S V A Q N K L L T P C D W E I 17

147 N K L L T P c D w E I L T K V 17

181 N Q D R K N R A A N P A I A I 17

186 N R A A N P A I A I T F E Q L 17

207 W G T V N N H Q D F E M M P L 17

49 W P Q E P Q A H G V A P V Q H 16

64 K A A L P S N V N E S P L Q F 16

78 F I I R Q A R L A G D L D A W 16

79 I I R Q A R L A G D L D A W Q 16

86 A G D L D A W Q F A V V L Q P 16

91 A W Q F A V V L Q P P R Q Q G 16

127 V G Q Y G P N S P F I R S L L 16

137 I R S L L Q S V A Q N K L L T 16

141 L Q S V A Q N K L L T P C D W 16

144 V A Q N K L L T P c D W E I L 16

146 Q N K L L T P c D W E I L T K 16

160 K V T L S P S Q F L Q F K T W 16

165 P S Q F L Q F K T W W T D E A 16

183 D R K N R A A N P A I A I T F 16

184 R K N R A A N P A I A I T F Ξ 16

204 G G Q W G T V N N H Q D F E M 16

60 P V Q H K A A L P S N V N E S 15

67 L P S N V N E S P L Q F I I R 15

73 E S P L Q F I I R Q A R L A G 15

92 W Q F A V V L Q P P R Q Q G G 15

106 G A H Q A V W Ξ P F S F K L L 15

135 P F I R S L L Q S V A Q N K L 15

208 G T V N N H Q D F E M M P L N 15

TABLE XXIV - 74P3B3, clone B

(frame 3) HLA-DRB1*0301

(DR17) 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Seq. ID Num .

116 S F K L L K D L K A A V G Q Y 31

138 R S L L Q S V A Q N K L L T P 27

190 N P A I A I T F E Q L L G I G 26

82 Q A R L A G D L D A W Q F A V 22

158 L T K V T L S P S Q F L Q F K 22

94 F A V V L Q P P R Q Q G G A H 20

108 H Q A V W E P F S F K L L K D 20

147 N K L L T P C D W E I L T K V 20

160 K V T L S P s Q F L Q F K T w 20

213 H Q D F E M P L N K F A I A 20

36 E s Q I C P V S R M N E L W P 19

76 L Q F I I R Q A R L A G D L.D 19

134 S P F I R S L L Q S V A Q N K 19

73 E s P L Q F I I R Q A R L A G 18

95 A V V L Q P P R Q Q G G A H Q 18

39 I C P V S R N E L W P Q E P 17

64 K A A L P S N V N E S P L Q F 17

112 W E P F s F K L L K D L K A A 17

148 K L L T P C D W E I L T K V T 17

177 D Ξ A Q N Q D R K N R A A N P 17

198 E Q L L G I G G Q W G T V N N 17

19 P E W P P P I K Q c S L E P W 16

56 H G V A P V Q H K A A L P S N 16

130 Y G P N s P F I R S L L Q S V 16

209 T V N N H Q D F Ξ M M P L N K 16

74 S P L Q F I I R Q A R L A G D 15

91 A W Q F A V V L Q P P R Q Q G 15

125 A A V G Q Y G P N S P F I R S 15

139 S L L Q S V A Q N K L L T P C 15

152 P C D W E I L T K V T L s P S 15

TABLE XXV - 74P3B3, clone B

(frame 3) HLA-DRB1*0401

(DR4Dw4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

165 P S Q F L Q F K T w W T D E A 28

55 A H G V A P V Q H K A A L P S 26

134 S P F I R S L L Q s V A Q N K 26

138 R S L L Q S V A Q N K L L T P 26

30 L E P W R S E S Q I C P V S R 22

46 N E L W P Q E P Q A H^' G V A P 22

91 A W Q F A V V L Q P P R Q Q G 22

109 Q V W E P F S F K L L K D L 22

114 P F S F K L L K D L K A A V G 22

152 P C D W E I L T K V T L S P S 22

168 F L Q F K T W w T D E A Q N Q 22

171 F K T W w T D E A Q N Q D R K 22

172 K T W W T D E A Q N Q D R K N 22

204 G G Q W G T V N N H Q D F E M 22

27 Q C S L E P w R S E S Q I C P 20

36 E S Q I C P V S R M N E L W P 20

73 E S P L Q F I I R Q A R L A G 20

82 Q A R L A G D L D A Q F A V 20

108 H Q A V E P F S F K L L K D 20

116 S F K L L K D L K A A V G Q Y 20

155 W E I L T K V T L S P S Q F L 20

158 L T K V T L S P S Q F L Q F K 20

192 A I A I T F E Q L L G I G G Q 20

198 E Q L L G I G G Q W G T V N N 20

19 P E W P P P I K Q C S L E P W 18 TABLE XXV - 74P3B3 , clone B

(frame 3 ) HLA-DRB1*0401

(DR4Dw4 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

61 V Q H K A A L P S N V N E S P 18

74 S P L Q F I I R Q A R L A G D 18

85 A G D L D A W Q F A V V L Q 18

130 Y G P N S P F I R S L L Q S V 18

131 G P N S P F I R S L L Q S V A 18

135 P F I R S L L Q S V A Q N K L 18

139 S L L Q S V A Q N K L L T P C 18

151 T P C D W E I L T K V T L S P 18

162 T L S P s Q F L Q F K T W W T 18

182 Q D R K N R A A N P A I A I T 18

188 A A N P A I A I T F E Q L L G 18

18 H P E w P P P I K Q C S L E P 16

89 L D A w Q F A V V L Q P P R Q 16

133 N S P F I R S L L Q S V A Q N 16

194 I T F E Q L L G I G G Q W G 16

42 V S R M N E L W P Q E P Q A H 14

64 K A A L P S N V N E S P L Q F 14

68 P S N V N Ξ S P L Q F I I R Q 14

76 L Q F I I R Q A R L A G D L D 14

77 Q F I I R Q A R L A G D L D A 14

86 A G D L D A W Q F A V V L Q P 14

93 Q F A V V L Q P P R Q Q G G A 14

94 F A V V L Q P P R Q Q G G A H 14

95 A V V L Q P P R Q Q G G A H Q 14

117 F K L L K D L K A A V G Q Y G 14

120 L K D L K A A V G Q Y G P N S 14

137 I R S L L Q S V A Q N K L L T 14

146 Q N K L L T P C D W E I L T K 14

147 N K L L T P C D W E I L T K V 14

190 P A I A I T F E Q L L G I G 14

197 F E Q L L G I G G Q W G T V N 14

200 L L G I G G Q W G T V N N H Q 14

207 W G T V N N H Q D F E M M P L 14

TABLE XXVI - 74P3B3, clone B (frame 3) HLA-DRB1*1101 15 - ' mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

55 A H G V A P V Q H K A A L P S 26

18 H P Ξ W P P P I K Q C S L E P 24

152 P C D E I L T K V T L S P S 24

114 P F S F K L L K D L K A A V G 23

194 A I T F E Q L L G I G G Q W G 22

36 E S Q I C P V s R M N E L w P 20

73 E S P L Q F I I R Q A R L A G 20

76 L Q F I I R Q A R L A G D L D 20

94 F A V V L Q P P R Q Q G G A H 20

155 W E I L T K V T L S P S Q F L 20

117 F K L L K D L K A A V G Q Y G 19

134 S P F I R S L L Q S V A Q N K 19

39 I C P V S R M N E L w P Q E P 18

42 V S R M N E L W P Q E P Q A H 18

75 P L Q F I I R Q A R L A G D L 18

89 L D A W Q F A V V L Q P P R Q 18

197 F E Q L L G I G G Q w G T V N 18

91 A W Q F A V V L Q P P R Q Q G 17 TABLE XXVI - 74P3B3, clone B

(frame 3) HLA-DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

213 H Q D F E M M P L N K F A I A 17

13 A P H W D H P E W P P P I K Q 16

30 L E P R S E S Q I c P V s R 16

46 N E L W P Q Ξ P Q A H G V A P 16

113 E P F S F K L L K D L K A A V 16

168 F L Q F K T W W T D E A Q N Q 16

177 D E A Q N Q D R K N R A A N P 16

204 G G Q w G T V N N H Q D F E M. 16

48 L W P Q E P Q A H G V A P V Q^' 15

90 D A W Q F A V V L Q P P R Q Q 15

24 P I K Q c S L Ξ P W R S E S Q 14

26 K Q c S L E P W R S E S Q I C 14

64 K A A L P S N V N E S P L Q F 14

116 S F K L L K D L K A A V G Q Y 14

120 L K D L K A A V G Q Y G P N S 14

154 D W E I L T K V T L S P S Q F 14

138 R S L L Q S V A Q N K L L T P 13

2 F K T K K G L Ξ E Q S A P H W 12

58 V A P V Q H K A A L P S N V N 12 95 A V V L Q P P R Q Q G G A H Q 12 105 G G A H Q A V W E P F S F K L 12 121 K D L K A A V G Q Y G P N S P 12 124 K A A V G Q Y G P N S P F I R 12 141 L Q S V A Q N K L L T P C D W 12 172 K T W W T D E A Q N Q D R K N 12 http://www.b_ni-heidelberg.coni/syfpeithi

TABLE XXIII - 83P4B8 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1100 D W L I T K L K G Q V S Q E T 35

1194 M E K L V K L S G s H L T P L 35

89 I I G L L M L E A H H F P G P 34

170 D Q Q Y V I Q H T s M F K D V 34

261 S G E L R H V E G T I I L H I 34

270 T I I L H I V F A I K L D Y E 34

338 F K D L Q L L Q G s K F L Q N 34

544 L K N F K V L G S L s S S Q C 34

214 V Y Q L L V L S S K G S R K S 33

384 E L G F I L M D S Y G P K K V 33

456 I S H F L D L L S N I V M Y A 32

1026 K S L M N L L F S L H V S Y K 32

1131 E K A I I M Q L G T L L T F F 32

127 L Ξ L L P I I L T A L A T K K 31

324 D Q V L D L L K T s V V K S F 31

410 L S R P N Q H A C K L G A N 31

1000 S P Q F V Q M L S W T S K I C 31

1071 H F A I V N L R T A A P T V C 31

70 T C C I Q L V E S G D L Q K E 30

83 K Ξ I V S E I I G L L L E A 30

227 K S V L E G I I A F F S A L D 30

347 S K F L Q N L V P H R S Y V S 30

442 E Q V L N R V V T R A S S P I 30

765 K N R F E D I L s L F M C Y K 30

807 M K F V S S L L T A L F R D S 30 TABLE XXIII - 83P4B8 HLA- DRBl*010l 15 - mers

Seq. ID

Pos ^• 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

253 L L D V V T V P S G E L R H V 29

982 A L L L V T V L T S L S K L L 29 1260 Y E K F L I H L s K K S K V N 29

250 G D E L L D V V T V P S G E L 28

974 E E D F N S K E A L L L V T V 28 1069 T N H F A I V N L R T A A P T 28

190 E V E F V V E K A L S M F S K 27

415 N Q H A C K L G A N I L L E T 27

494 Q T V Q R L L K A V Q P L L K 27

524 A M F A N Q L D A R K S A V A 27

598 Q A D V R L M L Y E G F Y D V 27

901 G K S I S L L c L E G L Q K I 27 1084 V C L L V L S Q A E K V L E E 27 1142 L T F F H E L V Q T A L P S G 27 1167 C K- M Y T T L T A L V R Y Y L 27

199 L S M F S K M N L Q E I P P L^' 26

379 T Q G L V E L G F I L M D S Y 26

393 Y G P K K V L D G K T I E T S 26

482 T E A F D Y L S F L P L Q T V 26

516 C L I L V L R K A M F A N Q L 26

538 A G F L L L L K N F K V L G S 26

641 L L P P L K L D A C I L T Q G 26

726 S A D F S Q S T S I G I K N N 26

753 I E Y N F S I S S F S K N R 26

989 L T S L S K L L E P S S P Q F 26

1030 N L L F S L H V S Y K S P V I 26

1212 F I S Y V Q N K S K S L N Y T 26

2 D Q K I L S L A A E K T A D K 25

26 E G D L T N L L Q N Q A V K G 25 35 N Q A V K G K V A G A L L R A 25

102 G P L L V E L A N E F I S A V 25

131 P I I L T A L A T K K E N L A 25

180 M F K D V P L T A E E V E F V 25

202 F S K M N L Q E I P P L V Y Q 25

217 L L V L S S K G s R K S V L E 25

224 G S R K S V L E G I I A F F S 25

278 A I K L D Y E L G R E L V K H 25

323 Q D Q V L D L L K T S V V K S 25

387 F I L M D S Y G P V L D G 25

^Kι^K

421 L G A N I L L E T F'K I H E M 25

457 S H F L D L L S N I V M Y A P 25

485 F D Y L S F L P L Q T V Q R L 25

572 H S H Y N S V A N ,E T F C L E 25

606 Y E G F Y D V L R R N S Q L A 25

718 L E D F E L D K S A D F S Q S 25

741 I S A F L V M G V C E V L I E 25

781 L S D I L N E K A G K A K T K 25

811 S S L T A L F R D S I Q S H 25

903 S I S L L c L E G I Q K I F S 25

925 P K I Q Q F L R A L D V T D K 25

948 D V S V T Q R T A F Q I R Q F 25

990 T S L s K L L E P S S P Q F V 25 1081 A P T V c L L V L S Q A E K V 25 1170 Y T T L T A L V R Y Y L Q V C 25 1176 L V R Y Y L Q V c Q S S G G I 25 1202 G S H L T P L c Y S F I S Y V 25 1233 P A A V A T A M A R V L R E-T 25 1270 K S K V N L M Q H M K L S T S 25 TABLE XXIII - 83P4B8 HLA- DRB1* 0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1276 M Q H K L S T S R D F K I K 25 18 Q E F L Q T L R E G D L T N L 24 36 Q A V K G K V A G A L L R A I 24 44 G A L L R A I F K G S P C S E 24 62 T L R R R K I Y T C C I Q L V 24 86 V S E I I G L L M L E A H H F 24 101 P G P L L V E L A N E F I S A 24 116 V R E G S L V N G K S L E L L 24 141 K E N L A Y G K G V L S G Ξ E 24 177 H T S M F K D V P L T A E E V 24 188 A E E V E F V V E K A L S M F 24 192 E F V V E K A L S M F S K M N 24 207 L Q E I P P L V Y Q L L V L S 24 211 P P L V Y Q L L V L S S K G S 24 319 I Q R F Q D Q V L D L L K T S 24 372 V H S W D H V T Q G L V E L G 24 463 L S N I V Y A P L V L Q S C 24 487 Y L S F L P L Q T V Q R L L K 24 500 L K A V Q P L L K V S M S M R 24 514 R D C L I L V L R K A M F A N 24 515 D C L I L V L R K A M F A N Q 24 543 L L K N F K V L G S L S S S Q 24 585 L E I M D S L R R C L S Q Q A 24 588 D S L R R C L S Q Q A D V R 24 631 L K Q F Y E P K P D L L P P L 24 636 E P K P D L L P P L K L D A C 24 648 D A C I L T Q G D K I S L Q E 24 856 T G H V S G P D G Q N P E K I 24 906 L L C L E G L Q K I F S A V Q 24 964 R S L L N L L S S Q E E D F N 24 1146 H E L V Q T A L P S G S C V D 24 1180 Y L Q V C Q S S G G I P K N 24 1283 T S R D F K I K G N I L D M V 24 32 L L Q N Q A V K G K V A G A L 23 118 E G S L V N G K S L E L L P I 23 126 S L E L L P I I L T A L A T K 23 158 K Q L I N T L c S G R W D Q Q 23 210 I P P L V Y Q L L V L S S K G 23 299 G D S N N N L S P F S I A L L 23 307 P F S I A L L L S V T R I Q R 23 339 K D L Q L L Q G S K F L Q N L 23 350 L Q N L V P H R S Y V S T M I 23 438 Q E I L E Q V L N R V V T R A 23 445 L N R V V T R A s s p- I S H F 23 446 N R V V T R A S S P I S H F L 23 470 A P L V L Q S c s s K V T E A 23 488 L S F L P L Q T V Q R L L K A 23 519 L V L R K A M F A N Q L D A R 23 582 T F C L E I M D S L R R C L S 23 612 V L R R N s Q L A N S V M Q T 23 680 N T V I P L Q Q G E E E Ξ E E 23 703 D D I L Ξ S I T N R M I K S E 23 734 S I G I K N N I S A F L V M G 23 735 I G I K N N I s A F L V M G V 23 778^' Y K K L S D I L N E K A G K A 23 798 N K T S D S L L S M K F V S S 23 842 A V N V A L Q K V Q Q L K E T 23. 882 L W R Y T S I P T S V E E S G 23 TABLE XXIII - 83P4B8 HLA- DRB1*0101 15 - mers

9 0 Seq. ID

Pos . 1 2 3 4 5 6 7 8 1 2 3 4 5 score Num.

980 K E A L L L V T V L T S L S K 23

983 L L L V T V L T S L S K L L E 23

985 L V T V L T S L S K L L E P S 23

1085 C L L V L S Q A E K V L E E V 23

1096 L E E V D W L I T K L K G Q V 23

1122 Q A T L P N Q P V E K A I I M 23

1132 K A I I M Q G T L L T F F H 23

1163 L K D L C K M Y T T L T A L V 23

1179 Y Y L Q V C Q S S G G I P K N 23

110 N E F I S A V R E G S L V N G 22

123 N G K s L E L L P I I L T A L 22

242 K Q H N E E Q S G D E L L D V 22

357 R S Y V S T M I L E V V K N S 22

361 S T M I L E V V K N S V H S W 22

398 V L D G K T I E T S P S L S R 22

407 s P S L s R M P N Q H A C K L 22

429 T F K I H E M I R Q E I L E Q 22

437 R Q E I L E Q V L N R V V T R 22

460 L D L L S N I V M Y A P L V L 22

462 L L S N I V M Y A P L V L Q S 22

468 M Y A P L V L Q S c S s K V T 22

527 A N Q L D A R K S A V A G F L 22

531 D A R K S A V A G F L L L L K 22

547 F K V L G S L S S S Q C S Q S 22

592 R R C L S Q Q A D V R L M L Y 22

609 F Y D V L R R N S Q L A N S V 22

621 N S V M Q T L L S Q L K Q F Y 22

655 G D K I S L Q E P L D Y L L c 22

665 D Y L L C C I Q H C L A W Y K 22

702 L D D I L E S I T N R M I K s 22

743 A F L V M G V c E V L I E Y N 22

751 E V L I E Y N F S I S s F s K 22

793 K T K M A N K T S D S L L s M 22

801 S D S L L S M K F V s S L L T 22

804 L L S M K F V S S L L T A L F 22

826 Q E S L S V L R S S N E F M R 22

871 F Q N L C D I T R V L L W R Y 22

878 T R V L L W R Y T S I P T S V 22

909 L E G L Q K I F s A V Q Q F Y 22

963 Q R S L L N L L s S Q E E D F 22

992 L S K L L E P S s P Q F V Q M 22

993 S K L L E P S s P Q F V Q M L 22

1034 S L H V S Y K s P V I L L R D 22

1042 P V I L L R D L s Q D I H G H 22

1061 D Q D V E V E K T N H F A I V 22

1112 Q E T L S Ξ Ξ A s S Q A T L P 22

1135 I Q L G T L L T F F H E L V 22

1144 F F H E L V Q T A L P S G S C 22

1166 L C K Y T T L T A L V R Y Y 22

1252 N L I F A I E Q Y E K F L I H 22

1273 V N L Q H M K L S T S R D F 22

79 G D L Q K Ξ I V S E I I G L L 21

139_. T K K E N L A Y G K G V L S G 21

155 E C K K Q L I N T L C s G R W 21

274 H I V F A I K L D Y E L G R E 21

305 L S P F S I A L L L s V T R I 21

497 Q R L L K A V Q P L L K V S M 21

513 M R D C L I L V L R K A M F A 21 o^j ω ω tυ i W O Ol

» i co c.

t¹ IT •TJ W < H D ι-_ IT IT > tr¹ tfl cn cn Ω O n ω o H tr¹ cn rt ≥! tr¹ < W < < < TJ tfl > IT H rt « cn α tfl < cn H Ω O Ω < « rt H » < α •fl H a a CO < co TI < a IT H Ω tfl O « Ω ιθ cn ιθ n rt ≥S tfl < > » « IT tfl tr IT Ω lO Ifl > tfl IT « tfl IT 2 en IT tr O TI 3 a •fl ri H CO CO 3 to

Ω IT H tfl H •fl TJ 2 H a a 3 <. D tfl tT H cn Ω •fl IT « tfl cn W rt f Ω cn cn o Ω < IT rt IO S « ≥i cn IT cn IT « cn > > IT w a ta •fl « Ω H Ω > α t cn tr > Ω < •fl tr¹ •fl H Ω •fl tr* cn IT ri f •fl H t* IT < H S •fl f •fl K Ά IT rt f tr tr κ| > J TJ •A Ω s •fl •A -3 fe! en jo IT Ά a tr •fl TI iO K ^ td t ω

« « IT β τι ω H « tr¹ W Ti tfl tfl O Ω t_J cn H TJ td Ω " f Ω H Ω « tr¹ W IT H a H IT ta tfl fd rt < n W w < CO O cn CO lO H t f. O O en « UI H I

*

•fl 1-3 tT H O O rt Cn Ω H t*. ι-3 IT tr¹ cn « > » O H H « f £3 tr* cn O H tr H ta H ?? Ω H « * cn « « < co ta co tr Ω H Ω w « σ\ o

H

IT O tr¹ H Ω IT H < tr¹ TJ W H tfl tT r* - tr¹ tfl C tfl « cn tfl •fl IT rt tr rt IT tr IT > Ω •fl IT tfl tr O tfl rt CO a ≥! 3 O H < ιO ta o o tr -α o

O < cn tT tfl D tfl TJ IT tfl f 8 Ω H td O < 3 α Ω « ≥i tfl H « IT S ιO cn X <! rt H a IT fti « tr tr * cn IT » rt ιθ Ω cn TI < lO j o a tfl CO oo 1 a < < M f « < f > f tr¹ tr* < td tfl lO tfl J f tfl ≥i -2i IT¹ S < « H IT < H tr > •fl IT H tfl rt « UJ 3 Ω H IT •fl rt cn rt •d to rt cn •fl TI cn σ VD in

IT « rt H td ιO tfl H ι-3 IT < TJ tT cn wo w a « rt O rt •fl H tfl α tr¹ -3 r ιτ < < CO IT « rt w rt > tfl o ≥I « T rt tfl H tr o 3 IT tfl iO iO tfl IT H o CO to < K W •fl > P. TJ tfl Ω 3 tfl W « tr¹ O « Ω cn CO α n Ω •fl tfl o ^ •fl ω > Ω a tr . S W tr tr CO rt α •d tr cn H < Ω tr tfl Pd « cn tr H 3 t ro

TI b_ H •fl < a <! tfl f*. H tT TJ IT H H > O « H W H •fl t→ ω W < Ω CO > « rt CO IT CO TJ TJ cn IT H cn tr H IT co IT IT ω TI tr CO H < a to m a « F tfl tfl < IS tfl H > F tfl H « σ « « tfl •fl <! cn •fl M ϊβ l→ co •fl H Ω « J tr w H » lO Ά cn < Ά f O O CO tfl o tr cn tr IT ι_ IT Ifl w t td α td H Ω tfl tfl < SJ F a IT > Ω rt Ω « CO f Ω cn cn CO t¹ IT fd cn IT « * « ?. CO « n io « TJ O cn IT tfl » cn IT < < IT < Ω cn IT IO w

CO F •fl ?? H_ « tfl tT ι-3 tfl <_ K F n ?. tr¹ r cn tfl Ω •fl α n co Ω D tr » tfl cn a ω S3 tr < H tr tr tfl α tr Ω « cn < tr a cn tfl a a CO cn cα

H H H H H H H H H H H H H t to t t to to t t to t to to to to t to to to t to t t n

CO 00 00 ω 00 CO vo vo vo O vo vo o o o o o o o o o o o o o o o o H H H H H H o

CO

3 "

' H α

TABLE XXIII - 83P4B8 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

360 V S T M I L E V V K N S V H S 18 376 D H V T Q G L V E L G F I L M 18 399 L D G K T I E T S P S L s R M 18 433 H E I R Q E I L E Q V L N R 18 461 D L L s N I V M Y A P L V L Q 18 466 I V M Y A P L V L Q S C S S K 18 496 V Q R L L K A V Q P L L K V S 18 510 S S M R D C L I L V L R K A 18 536 A V A G F L L L L K N F K V L 18 539 G F L L L L K N F K V L G S L 18 600 D V R L M L Y E G F Y D V L R 18 607 E G F Y D V L R R N S Q L A N 18 628 L S Q L K Q F Y E P K P D L L 18 632 K Q F Y E P K P D L L P P L K 18 647 L D A C I L T Q G D K I s L Q 18 695 E E A F Y E D L D D I L E S I 18 696 E A F Y E D L D D I L E S I T 18 700 E D L D D I L E S I T N R M I 18 715 K S E L E D F E L D K S A D F 18 738 K N N I s A F L V M G V C E V 18 742 S A F L V M G V C Ξ V L I E Y 18 771 I L S L F M C Y K K L S D I L 18 782 S D I L N E K A G K A K T K M 18 790 G K A K T K M A N K T S D s L 18 799 K T S D S L L S M K F V S s L 18 835 S N E F M R Y A V N V A L Q K 18 836 N E F M R Y A V N V A L Q K V 18 838 F M R Y A V N V A L Q K V Q Q 18 849 K V Q Q L K E T G H V S G P D 18 868 E K I F Q N L C D I T R V L L 18 876 D I T R V L L w R Y T S I P T 18 877 I T R V L L W R Y T s I P T S 18 919 V Q Q F Y Q P K I Q Q F L R A 18 920 Q Q F Y Q P K I Q Q F L R A L 18 922 F Y Q P K I Q Q F L R A L D V 18 933 A L D V T D K E G E E R Ξ D A 18 956 A F Q I R Q F Q R S L L N L L 18 959 I R Q F Q R S L L N L L S S Q 18 960 R Q F Q R S L L N L L S S Q E 18 978 N S K E A L L L V T V L T S L 18 1001 P Q F V Q M L S W T S K I C K 18 1043 V I L L R D L S Q D I H G H L 18 1063 D V E V E K T N H F A I V N L 18 1072 F A I V N L R T A A P T V C L 18 1127 N Q P V E K A I I M Q L G T L 18 1141 L L T F F H Ξ L V Q T A L P S 18 1191 P K N M E K L V K L S G S H L 18 1192 K N M E K L V K L S G S H L T 18 1239 A M A R V L R E T K P I P N L 18 1262 K F L I H L S K K S K V N L M 18 1264 L I H L S K K S K V N L M Q H 18 1301 D G E D E N Ξ E G T A S E H G 18 6 L S L A A E K T A D K L Q E F 17

29 L T N L L Q N Q A V K G K V A 17 30 T N L L Q N Q A V K G K V A G 17 39 K G K V A G A L L R A I F K G 17 41 K V A G A L L R A I F K G S P 17 52 K G S P C S E E A G T L R R R 17 H μ H

0 α

IT H « a rt < tfl α ^h_ a_ CO IT t*i D a D Ω td KD <! n > tr « tr P. cn < IT Ω IT « rt a < td D H tfl IT KD tfl cn tfl < Ω Ω Ω tfl TJ H cn KD n cn IT n cn rt > •fl > CO < cn Ά ω Ά cn < 3 a Ω td tr a α tr > CO tr tfl •fl < CO CO a « tfl « Ω t) CO Ω < « Ifl tfl i cn < •fl IT Ω tfl pi t tfl a TI •fl td « < t^l O > a td ω < IT * tr Ω H Ω H tr tfl tfl > to « < « a o TJ •fl < IT a tr tfl tr rt < Ω tr •A tfl cn IT td H IT IT ta ω α tfl tr < Ω tfl IT IT td <J < IT tfl α tr ω IT > co tfl « IT IT 3 rt td 3 < o <! td < tr a « < CO tr < tr α < O tfl tr < Ω tr < fl cn < tr td ω to H « O π rt tr ω cn o •A ω t co co Ω IT < W > rt IT 2 •fl * ω J rt IT 2 td α « TJ Pi « Ω J rt Ifl Ω < pi tfl a Ω ta 2 Ω < Π

* f en w < IT < ιθ « 3 H rt IT 3 * •A tr tfl -3 Ki o π 3 3 tr tfl <. a O cn IT •A < a td tfl IT < IT •fl 3 tfl i IT Ω cn < tr IT IT σι o co tr tr tr tfl IT rt tT rt >fl < td iO IT t*J 3 H tr K: Ω tfl cn tr > td cn IT rt td cn KJ iO •fl cn Ω IT tfl tr a •d tr •fl •fl i ιo IT « tr td tfl tr Ifl lO |f td 3 •A a Cn < g o K O K; cn tr rt •fl « Ω tfl Ifl « o Ω tr tfl < α 3 H •fl td < -< < IT Pi KD W IT Ω CO o cn cn IT TJ cn < tfl a > 2 TI σ a π o « tr tr o o o > a Ά < rt K o a « •fl td H > Ω td < α <! Ω O IT Ό ! iO <! < td < Ω < •fl « tr H IT a Ω H a IT in rt rt tr tr « !d tr rt tr rt j tr KD td ^'tr co * P. a •A rt H Hi td 3 Ω tr a ω «! rt TJ ι^ IO f IT Ω Ω i ta •fl tfl < t> •fl 3 CO a tfl Ω ϋ_ Ά a tfl Ω o tr < IT co tfl IT co cn ?. « a tfl O >fl 3 « •A td <! cn σ Ω α •fl Ω 3 tr cn fd « 3 Ω KD IT O KD a tr tr rt O ω a 3 rt IT tr « tr •A > IT 3

CD

TI Ω lO •A tr D tfl tr ω Ά H tfl ω cn td IT IT cn IT H IT < IT cn tr tr IT cn CO TJ > f. H CO IT tr CO KD tτ, H td < « tr •fl tr rt IT cn < CO J a tr t cn a IT O O 3 TJ O ιo 10 IO rt tr H O IT a CO H o TI KD IT CO IT tr < H 3 Ά H O < IT « IT cn a Ω w w « a TI o tr o cn Ω > TJ tr a Ω a 3 ιO IT H tr a tr CO « tfl H KD Ω tr cn o co rt « o cn tt tr ta td KD td t→ tr IT td tr cn CO IT tfl •fl KD < a D < a tr < cn a o Ifl « cn tr tfl Ω < J Ά IT _^

TJ < a a IT Ά tr tfl t Ά a o < cn H rt cn D TJ td KD td tr td < O TJ a TJ a < tr Ω rt a cn Ω tr o is Ω a « 3 Ω tr Pi P3 IT J ta tπ

H H P H H H H H H H H H H H H H H - H H H H M H H H H - O -J -J -O. -0 -0 -J -O. -0 -0 -J -0 o

a

H α

TABLE XXIII - 83P4B8 HLA- DRB1*0101 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 Seq. ID

4 5 score Num.

1130 V E K A I I M Q L G T L L T F 17 1133 A I I M Q L G T L L T F F H E 17 1148 L V Q T A L P s G S c V D T L 17 1160 D T L L K D L c K M Y T T L T 17 1195 E K L V K L S G S H L T P L c 17 1197 L V K L S G S H L T P L c Y S 17 1207 P L C Y S F I S Y V Q N K S K 17 1225 Y T G E K K E K P A A V A T A 17 1226 T G E K K E K P A A V A T A M 17 1229 K K E K P A A V A T A M A R V 17 1243 V L R E T K P I P N L I F A I 17 1254 I F A I E Q Y Ξ K F L I H L s 17 1261 E K F L I H L S K K S K V N L 17 1278 H M K L S T S R D F K I K G N 17 1286 D F K I K G N I L D M V L R E 17

TABLE XXIV - 83P4B8' HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1159 V D T L L K D L C K M Y T T L 31

276 V F A I K L D Y E L G R E L V 29

624 M Q T L L S Q L K Q F Y E P K 29

718 L E D F E L D K S A D F S Q S 29

1295 D M V L R E D G E D E N E E G 29

564 V S Q V H V D V H S H Y N S V 28

695 E E A F Y E D L D D I L E S I 28

1046 L R D L S Q D I H G H L G D I 28

196 E K A L s M F S K M N L Q E I 27

294 K V G Q Q G D S N N N L S P F 27

496 V Q R L L K A V Q P L L K V S 27

515 D C L I L V L R K A M F A N Q 27

757 N F S I S s F S K N R F E D I 27

836 N E F M R Y A V N V A L Q K V 27

966 L L N L L s S Q E E D F N S K 27

1057 L G D I D Q D V E V E K T N H 27

1085 C L L V L S Q A E K V L E E V 27

1264 L I H L s K K S K V N L M Q H 27

272 I L H I V F A I K L D Y E L G 26

664 L D Y L L C C I Q H C L A W Y 26

871 F Q N L C D I T R V L L W R Y 26

912 Ii Q K I F S A V Q Q F Y Q P K 26

956 A F Q I R Q F Q R S L L N L L 26

188 A E E V E F V V E K A L S M F 25

506 L L K V S M S M R D C L I L V 25

584 C L E I M D S L R R C L S Q Q 25

594 C L S Q Q A D V R L M L Y E G 25

649 A C I L T Q G D K I S L Q E P 25

946 D A D V ^■s V T Q R T A F Q I R 25

1170 Y T T L T A L V R Y Y L Q V C 25

18 Q E F L Q T L R E G D L T N L 24

110 N E F I s A V R E G S L V N G 24

385 L G F I L M D S Y G P K.K V L 24

1042 P V I L L R D L S Q D I H G H 24

1142^' L T F F H Ξ L V Q T A L P S G 24

642 L P P L K L D A C I L T Q G D 23

89 I I G L L M L,E A H H F P G P 22

101 P G P L L V E L A N Ξ F I S A 22 TABLE XXIV - 83P4B8 HLA- DRB1*0301 (DR17 ) 15 - mers

Pos ^• 1 2 3 4 5 6 7 8 9 Seq. ID

0 1 2 3 4 5 score Num.

126 s L E L L P I I L T A L A T K 22

226 R K S V L E G I I A F F S A L 22

302 N N N L S P F s I A L L L S V 22

332 T S V V K S F K D L Q L L Q G 22

807 M K F V S S L L T A L F R D S 22

814 L T A L F R D s I Q S H Q E S 22

985 L V T V L T S L s K L L E P S 22

992 L S K L L E P s s P Q F V Q M 22

1028 L M N L L F S L H V S Y K S P 22

1131 E K A I I M Q L G T L L T F F 22

9 A A Ξ K T A D K L Q E F L Q T 21

21 L Q T L R E G D L T N L L Q N 21

22 Q T L R E G D L T N L L Q N Q 21

82 Q K E I V S E I I G L L M L E 21

118 E G S L V N G K s L Ξ L L P I 21

147 G K G V L S G E Ξ C K K Q L I 21

177 H T S M F K D V P L T A Ξ E V 21

204 K M N L Q E I P P L V Y Q L L 21

207 L Q E I P P L V Y Q L L V L S 21

463 L S N I V M Y A P L V L Q S C 21

510 S M S M R D c L I L V L R K A 21

538 A G F L L L L K N F K V L G S 21

539 G F L L L L K N F K V L G S L 21

598 Q A D V R L M L Y E G F Y D V 21

634 F Y Ξ P K P D L L P P L K L D 21

638 K P D L L P P L K L D A C I L 21

655 G D K I S L Q E P L D Y L L C 21

657 K I S L Q E P L D Y L L C C I 21

842 A V N V A L Q K V Q Q L K E T 21

986 V T V L T S L S K L L E P S s 21

35 N Q A V K G K V A G A L L R A 20

70 T C C I Q L V E S G D L Q K E 20

119 G S L V N G K s L E L L P I I 20

191 V E F V V E K A L S M F s K M 20

202 F S K M N L Q E I P P L V Y Q 20

253 L L D V V T V P S G E L R H V 20

338 F K D L Q L L Q G S K F L Q N 20

340 D L Q L L Q G S K F L Q N L V 20

361 S T M I L E V V K N S V H S W 20

488 L S F L P L Q T V Q R L L K A 20

508 K V S M S M R D c L I L V L R 20

514 R D C L I L V L R K A M F A N 20

540 F L L L L K N F K V L G S L S 20

610 Y D V L R R N S Q L A N s V M 20

710 T N R M I K S E L Ξ D F E L D 20

714 I K S E L E D F E L D K. .S A D 20

734 S I G I K N N I S A F L V M G 20

742 S A F L V M G V c E V L I E Y 20

764 S K N R F E D I L S L F M C Y 20

806 S M K F V S S L L T A L F R D 20

870 I F Q N L C D I T R V L L W R 20

874 L C D I T R V L L W R Y T S I 20

974 E E D F N S K E A L L L V T V 20

981 E A L L L V T V L T S L S K L 20

1020 E D A L F C K S II M N Ii L F S 20

1034 S L H V S Y K S P V I L L R D 20

1061 D Q D V E V E K T N H F A I V 20

1083 T V C L L V L S Q A E K V L E 20 TABLE XXIV - 83P4B8 HLA- DRB1*0301 (DR17 ) 15 - mers

Pos 1 2 3 4 Seq. ID

5 6 7 8 9 0 1 2 3 4 5 score Num.

1133 A I I M Q L G T L L T F F H E 20

1194 M E K L V K L S G s H L T P L 20

1270 K S K V N L M Q H M K L S T S 20

1276 M Q H M K L S T s R D F K I K 20

14 A D K L Q E F L Q T L R E G D 19

26 E G D L T N L L Q N Q A V K G 19

134 L T A L A T K K E N L A Y G K 19

215 Y Q L L V L S S K G S R K S V 19

235 A F F S A L D K Q H N E E Q S 19

270 T I I L H I V F A I K L D Y E 19

278 A I K L D Y E L G R E L V K H 19

282 D Y E L G R E L V K H L K V G 19

286 G R E L V K H L K V G Q Q G D 19

310 I A L L L S V T R I Q R F Q D 19

311 A L L L S V T R I Q R F Q D Q 19

316 V T R I Q R F Q D Q V L D L L 19

318 R I Q R F Q D Q V L D L L K T 19

323 Q D Q V L D L L K T S V V K S 19

327 L D L L K T S V V K S F K D L 19

334 V V K S F K D L Q L L Q G S K 19

382 L V E L G F I L M D S Y G P K 19

394 G P K K V L D G K T I E T S P 19

410 L S R M P N Q H A C K L G A N 19

423 A N I L L E T F K I H E M I R 19

500 L K A V Q P L L K V S M S M R 19

537 V A G F L L L L K N F K V L G 19

592 R R C L S Q Q A D V R L M L Y 19

602 R L M L Y E G F Y D V L R R N 19

616 N S Q L A N S V M Q T L L S Q 19

620 A N S V M Q T L L s Q L K Q F 19

631 L K Q F Y E P K P D L L P P L 19

711 N R M I K S E L E D F E L D K 19

802 D S L L S M K F V S S L L T A 19

828 S L S V L R s S N E F M R Y A 19

857 G H V S G P D G Q N P Ξ K I F 19

901 G K S I S L L C L E G L Q K I 19

924 Q P K I Q Q F L R A L D V T D 19

959 I R Q F Q R S L L N L L s s Q 19

1021 D A L F C K S L M N L L F s L 19

1086 L L V L S Q A E K V L Ξ E V D 19

1174 T A L V R Y Y L Q V C Q S s G 19

1262 K F L I H L S K K S K V N L M 19

97 A H H F P G P L L V Ξ L A N E 18

102 G P L L V E L A N E F I S A V 18

133 I L T A L A T K K E N L A Y G 18

135 T A L A T K K E N L A Y G K G 18

182 K D V P L T A Ξ E V E F V V E 18

198 A L S M F S K M N L Q E I P P 18

331 K T S V V K s F K D L Q L L Q 18

407 S P S L S R M P N Q H A C K L 18

429 T F K I H E M I R Q E I L Ξ Q 18

457 S H F L D L L s N I V M Y A P 18

480 K V T E A F D Y L S F L P L Q 18

482 T E A F D Y L s F L P L Q T V 18

490 F L P L Q T V Q R L L K A V Q 18

493 L Q T V Q R L L K A V Q P L L 18

585 L E I M D S L R R C L S Q Q A 18

628 L S Q L K Q F Y E P K P D L L 18 TABLE XXIV - 83P4B8 HLA- DRB1* 0301 (DR17 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

679 K N T V I P L Q Q G E E E E E 18

743 A F L V M G V C E V L I E Y N 18

749 V C E V L I E Y N F S I S S F 18

771 I L S L F M C Y K K L s D I L 18

778 Y K K L S D I L N E K A G K A 18

793 K T K M A N K T S D S L L s M 18

819 R D s I Q S H Q E S L S V L R 18

829 L S V L R S S N E F M R Y A V 18

844 N V A L Q K V Q Q L K E T G H 18

889 P T S V E E s G K K E K G K S 18

905 S L L C L E G L Q K I F S A V 18

919 V Q Q F Y Q P K I Q Q F L R A 18

932 R A L D V T D K E G E E R E D 18

967 L N L L S s Q E E D F N S K E 18

970 L S S Q E E D F N S K E A L L 18

1022 A L F C K S L M N L L F S L H 18

1043 V I L L R D L S Q D I H G H L 18

1099 V D W L I T K L K G Q V S Q E 18

1127 N Q P V E K A I I M Q L G T L 18

1139 G T L L T F F H E L V Q T A L 18

1156 G S C V D T L L K D L C K M Y 18

1197 L V K L S G S H L T P L C Y S 18

1209 C Y S F I S Y V Q N K S K S L 18

1213 I s Y V Q N K S K S L N Y T G 18

1220 s K s L N Y T G E K K Ξ K P A 18

1233 P A A V A T A M A R V L R E T 18

1240 M A R V L R E T K P I P N L I 18

1244 L R E T K P I P N L I F A I E 18

1250 I P N L I F A I E Q Y E K F L 18

1254 I F A I E Q Y E K F L I H L S 18

1261 E K F L I H L S K K S K V N L 18

1291 G N I L D M V L R E D G E D E 18

1298 L R Ξ D G E D E N E E G T A S 18

3 Q K I L S L A A E K T A D K L 17

17 L Q E F L Q T L R E G D L T N 17

44 G A L L R A I F K G S P C S E 17

74 Q L V E S G D L Q K E I V S E 17

103 P L L V Ξ L A N E F I s A V R 17

140 K K E N L A Y G K G V L S G E 17

164 L C s G R W D Q Q Y V I Q H T 17

181 F K D V P L T A E E V E F V V 17

274 H I V F A I K L D Y Ξ L G R Ξ 17

290 V K H L K V G Q Q G D s N N N 17

313 L L s V T R I Q R F Q D Q V L 17

319 I Q R F Q D Q V L D L L K T S 17

362 T M I L E V V K N S V H s w D 17

416 Q H A C K L G A N I L L E T F 17

434 E M I R Q E I L E Q V L N R V 17

438 Q E I L E Q V L N R V V T R A 17

489 S F L P L Q T V Q R L L K A V 17

525 M F A N Q L D A R K S A V A G 17

568 H V D V H S H Y N S V A N E T 17

580 N E T F C L E I M D S L R R C 17

599 A D V R L M L Y E G F Y D V L 17

625 Q T L L S Q L K Q F Y E P K P 17

672 Q H C L A W Y K N T V I P L Q 17

682 V I P L Q Q G E E E E E E E E 17

687 Q G E E E E E E E E A F Y E D 17 TABLE XXIV - 83P4B8 HLA- DRB1 *0301 (DR17 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

688 G E E E E E E E E A F Y E D L 17

699 Y E D L D D I L Ξ S I T N R M 17

703 D D I L E S I T N R M I K S E 17

811 S S L L T A L F R D S I Q S H 17

840 R Y A V N V A L Q K V Q Q L K 17

847 L Q K V Q Q L K E T G H V S G 17

916 F S A V Q Q F Y Q P K I Q Q F 17

973 Q E E D F N S K Ξ A L L L V T 17

989 L T S L S K L L E P S S P Q F 17

1004 V Q M L S W T S K I C K Ξ N S 17

1160 D T L L K D c K M Y T T L T 17

1187 S G G I P K N M E K L V K L s 17

1191 P K N M E K L V K L S G S H L 17

1237 A T A M A R V L R E T K P I P 17

1251 P N L I F A I E Q Y Ξ K F L I 17

1252 N L I F A I E Q Y E K F L I H 17

1272 K V N L M Q H M K L S T S R D 17

1280 K L S T S R D F K I K G N I L 17

1284 S R D F K I K G N I L D M V L 17

40 G K V A G A L L R A I F K G S 16

58 E E A G T L R R R K I Y T C C 16

109 A N E F I S A V R E G S L V N 16

237 F S A L D K Q H N E Ξ Q S G D 16

606 Y E G F Y D V L R R N S Q L A 16

608 G F Y D V L R R N S Q L A N S 16

617 S Q L A N S V M Q T L L S Q L 16

653 T Q G D K I S L Q E P L D Y L 16

732 S T S I G I K N N I S A F L V 16

772 L S L F M c Y K K L S D I L N 16

885 Y T S I P T S V E E S G K K Ξ 16

906 L L C L E G L Q K I F S A V Q 16

920 Q Q F Y Q P K I Q Q F L R A L 16

931 L R A L D V T D K E G E E R E 16

972 S Q E E D F N S K Ξ A L L L V 16

1032 L F S L H V S Y K S P V I L L 16

1054 H G H L G D I D Q D V E V E K 16

1114 T L S E E A S S Q A T L P N Q 16

1210 Y S F I S Y V Q N K S K s L N 16

1253 L I F A I E Q Y E K F L I H L 16

10 A E K T A D K L Q E F L Q T L 15

113 I S A V R E G S L V N G K S L 15

149 G V L S G E E C K K Q L I N T 15

219 V L S S K G S R K S V L E G I 15

234 I A F F S A L D K Q H N E E Q 15

326 V L D L L K T S V V K S F K D 15

376 D H V T Q G L V E L G F I L M 15

388 I^'L M D S Y G P K K V L D G K 15

430 F K I H E M I R Q Ξ I L E Q V 15

572 H S H Y N S V A N Ξ T F C L E 15

630 Q L K Q F Y E P K P D L L P P 15

747 M G V C Ξ V L I E Y N F S I S 15

761 S S F S K N R F E D I L S L F 15

801 S D S L L S M K F V s S L L T 15

815 T A L F R D S I Q S H Q E S L ' 15

818 F R D S I Q S H Q E s L S V L 15

860 S G P D G Q N P E K I F Q N L 15

955 T A F Q I R Q F Q R s L L N L 15

1012 K I C K E N S R E D A L F C K 15 H H H H H H H H H H H H H O O O O O vo o vo vo vo oD co oo co ω -j ^ ι w σι iπ Lπ Lπ ^ ι^ ιP' i^ _-. *» ⁾ _ _ to H H H H -J m m ui iP' U H' τi TJ

-o w ω μ a αi *' * - to _0 C0 Lπ H O -J _N J H -4 *. o σ> tO C0 l H VD VD lπ ιl^ rfi l-> t0 lπ Ul H VD to O H io

H CΛ O O rf^ lπ -J O 0 o oo to to en io cn to c cnn ω t τ» (Λ vo H θ θ H ι^ ω vo L Lπ H iπ co _-. cn o ~o jι t co rf^ ω o _-> t -J t to ω ιπ -j Λ ι^ σ» t vD cn o 0

01

IT pi D tr rt IT TJ « IT Α LT •fl td a cn tr < O α a IT td < •fl cn a tfl o O IT Ω < tfl tr Ω rt tfl H IT KD « tfl K IT H < H α rt H rt Ω tfl tfl < td < cn tr tr •fl CO tfl KD tfl O IT « Ω ϋ cn Ifl Ω CI KD tr a td KD tfl KD IT α K •fl ta TJ Ω Ω tr rt pi a Ω tfl « cn a a t td a « to o td rt rt H H tfl Ω a IT tr IT KD Ω a > •A tT cn « tfl H IT < H •fl Ω tfl •a •fl <j H < α tfl KD < IT tr Ω O cn •fl H td •fl Ω a a cn tfl H * tr ω

H

IT IT H IT < tr tr tr 3 < IT H tr tr < 3 IT 3 tr < tr IT 3 3 IT tr t IT tr < IT IT IT < IT tT < IT IT H tr IT •fl Ά •fl Ά •A •fl =3 Ά _=> i tr pi _^ * I tfl t rt tr 3 cn O tr cn tr a rt < td KD KD Ω a td Pi tr tfl Ω o O IT tT iO O l a tfl a <: IT IT tfl TJ o ^ > a c tfl ϋ « IT O in O

H Ω in

X

> rt o tfl -3 <! o td tr <! rt ιθ iO pi o > •fl t) H CO Ω rt CO IT IT « rt LT td td D IT rt α < « tfl IT a > tfl t) < σ o a CO n „ KD n tr Ά tr < tfl IT IT α σ IT r < •fl Α H > IT < H tfl H tr IT tr < > < IT > < < IT < < IT > IT < tr tfl IT < H IT < IT tr < α td 1 < α

< *fl Ω >-H CO tr Ά rt IT iO _ rt r < •fl cn IT rt o tr td pi IT < o ω cn < IT Pi TJ < tr IT tfl IT « < ιO IT td tr Ω IT rt < oo td

H td a rt o rt KD a cn cn cn rt td o co td KD a td cn a a a a cn td a td KD tfl a cn rt a rt CO rt cn cn rt a co o rt iO n td td CO co lO td o o

TI

H. tfl IT O p_. > Ω KD tr tr cn TJ < pi <! O tr tfl •A td Ω KD Ω •A Pi T) LT TJ t td Ω < * 3 fl Ω a ri >fl tr a td IT a Ω tfl tfl ω tr IT o tr tfl a α a c tr tr « < tr < > CO tr Pi co 2 IT tr IT « > IT tr < > < < tfl TJ Ω •fl IT •A α KD a •fl ω a H f Ω H in rt tr < rt pi pi tr < « cn V H KD IT KD LT H « H « CO <! 3 tr w 3 cn cn < < tr CO cn cn H tr i tr to 3 cn < < CO t a tr to tr

KD KD >fl rt Ω < Ω a CO IT ^■Pi -. O O S O O O Q CO * -3 KD O IT •fl « a CO i Pi tfl Ω td « rt CO KD < tr TJ TJ Ω IT iO co 3 ta IT ω co 3 t

3 CD

< rt Ά IT iO IT o Ω K tr LT tr K Ά td IT « cn tr « CO CO KJ •fl o Ω < > Ά TJ td CO a tfl « 3 « If Pi « ιo co « > tr KD IT < *> CD Hi Hi tfl

Ω > a TJ < tfl a P. tfl IT tr Ά « < a •A > •fl Ifl Pi -< > cn a cn o TJ tr > •fl < IT cn a * Ifl Ω tfl Ω H W a > Ω > Ω a in ta Lπ

01 01 t to to to to to to to t to t to to to to to to to t to to to t t to t to to to t t to t to to t t t to to t t t to t to to to to t to to o H o cn cn cn cn cn cn n n en n n cn cn cn cn n en n n cn cn n cn cn en en cn cn n en cn en cn cn cn n n cn cn cn cn o in o

I (D (D

CO cn « CD r-. CD

3 3

--l r-> i-' r-' -^r l-- l-^r t-- r^J - H H H H H H I-¹ M t-^r r-< r-< r-' r-¹

O U) OO CO >J s] _] Ji W W w to ι to. r H r H w O w O _l V_JO VO 00 C0 CO -J *> _O H H M to tO t tO H H o ^ o ! o_ ∞ oo ι -^j ι ι c ιn ιπ ιn *- ιlⁱ. ι^ ι to tθ H t troI ϊ to. !r. TJ

•j σ. cD io -j H co oo H o c cnn Φ*'■ O0o0 oO i tfπl iloo ¹ H-^ O O. l _ CO l VD OO _-. n in H <-. _f-. __. (_i _f-. ^c° l-^a ^^{J IJ1} t0 H V0 00 ~J -0 C0 00 00 O > H V0 ^{u tJ} ^ ^{ul u} to ιn ιn ιo en oo ιn o to o -θ ιt» in co o

-j H vo o o . to ιn ιf* to o H I O 01

to to to t to t to to to to to to to to to to t to to to to to to to t to to to t to to to t to to t t to to to to t to to t to to t to t t t to t t t to t o

O O o o o o o o o o o o o o o o o o o o o o to to t t to to to to to to to t t t t t to to to to t to to t to to t en n cn o n

cn M ft⁾

3

' H α

TABLE XXV - 83P4B8 HLA- DRB1* 0401 (DR4Dw4 ) 15 - mers

Seq. ID

Pos ^■ 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

211 P P L V Y Q L L V L s S K G S 20

227 K S V L E G I I A F F S A L D 20

230 L E G I I A F F S A L D K Q H 20

231 E G I I A F F S A L D K Q H N 20

237 F S A L D K Q H N E E Q S G D 20

261 S G E L R H V E G T I I L H I 20

269 G T I I L H I V F A I K L D Y 20

270 T I I L H I V F A I K L D Y E 20

278 A I K L D Y E L G R E L V K H 20

282 D Y E L G R E L V K H L K V G 20

287 R E L V K H L K V G Q Q G D S 20

290 V K H L K V G Q Q G D S N N N 20

302 N N N L S P F S I A L L L S V 20

307 P F S I A L L L s V T R I Q R 20

309 S I A L L L S V T R I Q R F Q 20

310 I A L L L S V T R I Q R F Q D 20

313 L L S V T R I Q R F Q D Q V L 20

316 V T R I Q R F Q D Q V L D L L 20

323 Q D Q V L D L L K T S V V K S 20

332 T S V V K S F K D L Q L L Q G 20

347 S K F L Q N L V P H R S Y V S 20

350 L Q N L V P H R s Y V S T M I 20

360 V S T M I L E V V K N S V H S 20

361 S T M I L E V V K N S V H S w 20

362 T M I L E V V K N S V H S D 20

369 K N S V H S W D H V T Q G L V 20

379 T Q G L V E L G F I L M D S Y 20

385 L G F I L M D S Y G P K K V L 20

395 P K K V L D G K T I E T S P S 20

407 S P S L S R M P N Q H A C K L . 20

424 N I L L E T F K I H E M I R Q 20

429 T F K I H E M I R Q E I L E Q 20

433 H E M I R Q E I L E Q V L N R 20

453 S S P I S H F L D L L s N I V 20

464 s N I V M Y A P L V L Q S C S 20

470 A P L V L Q S C S s K V T E A 20

471 P L V L Q S C S S K V T E A F 20

478 S S K V T fi A F D Y L S F L P 20

485 F D Y L S F L P L Q T V Q R L 20

493 L Q T V Q R L L K A V Q P L L 20

497 Q R L L K A V Q P L L K V S M 20

500 L K A V Q P L L K V S M S M R 20

506 L L K V S M S M R D C L I L V 20

518 I L V L R K A M F A N Q L D A 20

534 K S A V A G F L L L L K N F K 20

541 L L L L K N F K V L G S L S S 20

546 N F K V L G S L S S S Q C S Q 20

547 F K V L G S L S S S Q C S Q S 20

564 V S Q V H V D V H s H Y N S V 20

566 Q V H V D V H s H Y N S V A N 20

568 H V D V H S H Y N s V A N E T 20

584 C L E I M D S L R R C L S Q Q 20

588 M D. S L R R C L S Q Q A D V R 20

601 V R L M L Y E G F Y D V L R R 20

610 Y D V L R R N S Q L A N S V M 20

625 Q T L L S Q L K Q F Y E P K P 20

642 L P P L K L D A C I L T Q G D 20

661 Q E P L D Y L L C C I Q H C L 20 H H H H tO H H H o vo vo ι to ιπ rf* rf*

Ω B 3 rt a a α a Ω H a H α a > > a O α a a < a 3 H a a a a PJ >o a o -> ^ H cn Ω a a Co KD < 3 a a a B < a ω a a a D a α co π a Ω a B rt 3 KD rt 53 a a a a <J KD Ω Ω CO a cn KD rt a cn a iO a KD a cn H a < < a a cn a co σ B < 3 a a H cn α B a a α to a ω x a a a a D a a a XD a a a < a rt B α α X a a a a a O rt > a CO a a a Ω a cn n a > cn cn cn a α H α a Ω a a Ω B α α rt Ω ω H

* a < a < 3 a a < a a a H a < < H < < H a a < a H < a a a a a H H a a a H a a < a a < H a H H < < H H a H a < a a rf* o rf* 3 a < a Ω a a rt Ω a a rt a a Ω < a a a Ω a H a Ω KD cn a a a a a iO a B Ω a cn s a CO a co a __ a a Ω 3 cn a a a α H > a in o

H a a a rt a a rt a rt a Ω a a a a a a <: K D D a cn a 3 a rt a a a cn Ω a Ω a a a a <! rt CO a a CO a Ω a α B α TJ s Ω en __ a cn a rt 3 a a a KD a < a a a rt B o a a a a a a ω < a a a a p IT a a a < O a > a a a a a < a a a CQ H a Ω α a

Ω Ω co tr a t cn a o a < ιo t*

Ω a a a <. a α < < a a a < σ CO a a a cn a a r Ω a Ω rt KD cn a a a a 3 •A a a Ω a a a ιo a i α CO Ω rt rt a a a rt CO Ω s σ a rt a rt B KD O o < cn -3 B cn < O n < a iO a a Ω a CO o a cn a rt Ω 3 CO H B < cn a rt a D a ιo rf_* ω a cn > rt 3 CO a a 3 KD KD a -3 co a a < o α a O a rt a a a iO cn rt KD H KD a a a a co a If Ω Ω a < 3 α a Ω rt a o a a a Ω a a Ω <! a KD B < a KD > a B < H ω Ki a cn cn a rt B KD O a a a >o Ω a a a a o a a a K; n κ| a Ω a a a B < Ω H in

H rt a < rt rt cn to a a rt CO rt a a a B X KD a α a cn a cn a a a o K; cn H a a rt a 3 B co a a a CO a H < a co 3 rt B a to co a rt cn a > rt Ω rt B Ω a a a rt a rt < > rt < Ω o cn >^■ H a B a α B B < KD a H o co rt a a H a a a a a a B Ω <! H a a a ω

3 a K a a a rt CO a a a a a a a a H a a Ω KD a CO a o Ω rt a cn a Pi < Ω 3 α rt cn a cn cn κ| a 3 α a a B a -3 rf* (D Hi

< Ω a Ώ X < rt O a < a H rt a a < Ω a < a a a a a a a CO a a a a α a D > CO H a X a CO CO a α CO a a < Ω a CO 3 B KD in 01

01 to to to to to to to to to to t t to t to to t to to t t to to to t to t t t to to to to t to t to to to to to to t to to t t t to t to to t to t t to to r_ o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o

3

H α

TABLE XXV - 83P4B8 HLA- DRB1* 0401 (DR4DW4 ) 15 - mers

Seq. ID

Pos ■ 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1233 P A A V A T A M A R V L R E T 20

1261 E K F L I H L S K K S K V N L 20

1270 K S K V N L M Q H M K L S T S 20

1291 G N I L D M V L R E D G E D E 20

1295 D M V L R E D G E D E N E E G 20

22 Q T L R E G D L T N L L Q N Q 18

23 T L R E G D L T N L L Q N Q A 18

27 G D L T N L L Q N Q A V K G K 18

57 S E E A G T L R R R K I Y T C 18

62 T L R R R K I Y T C C I Q L V 18

74 Q L V E S G D L Q K E I V S E 18

79 G D L Q K E I V S E I I G L L 18

112 F I S A V R E G S L V N G K S 18

115 A V R E G S L V N G K S L E L 18

155 E C K K Q L I N T L C S G R 18

164 L C S G R D Q Q Y V I Q H T^' 18

168 R D Q Q Y V I Q H T s M F K 18

169 W D Q Q Y V I Q H T s M F K D 18

208 Q E I P P L V Y Q L L V L S S 18

262 G E L R H V Ξ G T I I L H I V 18

279 I K L D Y E L G R E L V K H L 18

284 Ξ L G R E L V K H L K V G Q Q 18

294 K V G Q Q G D S N N N L S P F 18

334 V V K S F K D L Q L Q G S K 18

337 S F K D L Q L L Q G s K F Q 18

343 L L Q G S K F L Q N L V P H R 18

354 V P H R S Y V S T M I L E V V 18

398 V L D G K T I E T S P s L S R 18

399 L D G K T I E T S P s L S R M 18

404 I E T S P S L S R M P N Q H A 18

421 L G A N I L L E T F K I H E M 18

430 F K I H E M I R Q E I L Ξ Q V 18

435 M I R Q E I L E Q V L N R V V 18

439 E I L E Q V L N R V V T R A S 18

443 Q V L N R V V T R A S S P I S 18

467 V M Y A P L V L Q S C S S K V 18

468 M Y A P L V L Q S C S S K V T 18

521 L R K A M F A N Q Ii D A R K S 18

525 M F A N Q L D A R K S A V A G 18

558 C S Q S L S V S Q V H V D V H 18

565 S Q V H V D V H S H Y N S V A 18

594 C L S Q Q A D V R L M L Y E G 18

613 L R R N S Q L A N.S V M Q T L 18

614 R R N S Q L A N S V M Q T L L 18

617 S Q L A N S V M Q T L L S Q L 18

622 S V M Q T L L S Q K Q F Y E 18

646 K L D A C I L T Q G D K I S L 18

700 E D L D D I L E S I T N R M I 18

722 E L D K S A D F S Q S T S I G 18

723 L D K S A D F S Q S T S I G I 18

731 Q S T S I G I K N N I s A F L 18

754 I E Y N F S I S S F s K N R F 18

786 N E K A G K A K T K M A N K T 18

803 S L L S M K F V S S L L T A L 18

825 H Q E S L S V L R S S N E F M 18

832 L R S S N E F M R Y A V N V A 18

870 I F Q N L C D I T R V L L W. R 18

876 D I T R V L L w R Y T s I P T 18 TABLE XXV - 83P4B8 HLA- DRB1*0401 (DR4DW4 ) 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 Seq. ID

1 2 3 4 5 score Num.

887 s I P T S V E E S G K K E K G 18

936 V T D K Ξ G E E R E D A D V S 18

942 E E R E D A D V S V T Q R T A 18

945 E D A D V S V T Q R T A F Q I 18

950 S V T Q R T A F Q I R Q F Q R 18

960 R Q F Q R s L L N L L s S Q E 18

971 S S Q E E D F N S K Ξ A L L L 18

979 S K E A L L L V T V L T S L S 18 \

991 S L S K L L E P S s P Q F V Q 18

997 Ξ P S S P Q F V Q M L S W T S 18

1039 Y K S P V I L L R D L S Q D I 18

1068 K T N H F A I V N L R T A A P 18

1075 V N L R T A A P T V c L L V L 18

1104 T K L K G Q V S Q E T L S E Ξ 18

1113 E T L S Ξ E A S S Q A T L P N 18

1119 A s S Q A T L P N Q P V E K A 18

1129 P V E K A I I M Q L G T L L T 18

1153 L P S G S c V D T L L K D L C 18

1206 T P L C Y s F I S Y V Q N K S 18

1230 K E K P A A V A T A M A R V L 18

1258 E Q Y Ξ K F L I H L S K K S K 18

1269 K K S K V N L M Q H M K L s T 18

920 Q Q F Y Q P K I Q Q F L R A L 17

17 L Q E F L Q T L R Ξ G D L T N 16

66 R K I Y T C C I Q L V Ξ S G D 16

97 A H H F P G P L L V E L A N E 16

170 D Q Q Y V I Q H T S M F K D V 16

178 T S M F K D V P L T A E E V E 16

199 L S M F S K M N L Q E I P P L 16

233 I I A F F S A L D K Q H N E E 16

280 K L D Y E L G R E L V K H L K 16

319 I Q R F Q D Q V L D L L K T S 16

335 V K S F K D L Q L L Q G S K F 16

356 H R S Y V S T M I L E V V K N 16

384 E L G F I L M D s Y G P K K V 16

427 L E T F K I H E M I R Q E I L 16

466 I V M Y A P L V L Q S C S s K 16

523 K A M F A N Q L D A R K s A V 16

603 L M L Y E G F Y D V L R R N S 16

631 L K Q F Y E P K P D L L P P L 16

674 C L A W Y K N T V I P L Q Q G 16

675 L A W Y K N T V I P L Q Q G E 16

696 E A F Y E D L D D I L E S I T 16

741 I S A F L V M G V c E V L I E 16

755 E Y N F S I S S F s K N R F E 16

760 I S S F S K N R F E D I L S L 16

806 s M K F V s S L L T A L F R D 16

835 s N E F M R Y A V N V A L Q K 16

838 F M R Y A V N V A L Q K V Q Q 16

868 E K I F Q N L C D I T R V L L 16

880 V L L W R Y T S I P T S V Ξ E 16

927 I Q Q F L R A L D V T D K E G 16

959 I R Q F Q R S L L N L L S S Q 16

1021 D A L F C K S L M N L L F S L 16

1098 E V D W L I T K L K G Q V S Q 16

1167 C K M Y T T L T A L V R Y Y L 16

1176 L V R Y Y L Q V C Q s S G G I 16

1222 s L N Y T G Ξ K K E K P A A V 16 TABLE XXV - 83P4B8 HLA- DRB1*0401 (DR4D 4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1252 N L I F A I E Q Y E K F L I H 16 217 L L V L S S K G S R K S V L E 15 445 L N R V V T R A S S P I S H F 15 527 A N Q L D A R K S A V A G F L 15 609 F Y D V L R R N S Q L A N S V 15 732 S T S I G I K N N I S A F L V 15 793 K T K M A N K T S D S L L S M 15 21 L Q T L R E G D L T N L L Q N 14 30 T N L L Q N Q A V K G K V A G 14 44 G A L R A I F K σ S P C S E 14 65 R R K I Y T C C I Q L V E S G 14 72 C I Q L V E S G D L Q K E I V 14 87 S E I I G L L M L Ξ A H H F P 14 89 I I G L L M L Ξ A H H F P G P 14 101 P G P L L V E L A N E F I S A 14 129 P I I L T A L A T K K E N 14 141 K E N L A. Y G K G V L S G E E 14 . 147 G K G V L S G E Ξ C K K Q L I 14 161 I N T L C S G R D Q Q Y V I 14 202 F S K M N Q E I P P L V Y Q 14 204 K M N L Q E I P P L V Y Q L L 14 210 I P P L V Y Q L L V L S s K G 14 216 Q L L V L S S K G S R K s V L 14 251 D E L D V V T V P S G E L R 14 256 V V T V P S G E L R H V E G T 14 264 L R H V E G T I I L H I V F A 14 268 E G T I I L H I V F A I K L D 14 272 I L H I V F A I K L D Y E L G 14 273 L H I V F A I K L D Y E L G R 14 276 V F A I K L D Y Ξ L G R E L V 14 286 G R E L V K H L K V G Q Q G D 14 292 H L K V G Q Q G D S N N N L S 14 326 V L D L L K T S V V K S F K D 14 331 K T S V V K S F K D L Q L L Q 14 338 F K D L Q L L Q G S K F L Q N 14 340 D L Q L L Q G S K F L Q N L V 14 341 L Q L L Q G S K F L Q N L V P 14 357 R S Y V S T M I L E V V K N S 14 365 E V V K N S V H S W D H V T 14 382 L V E L G F I L M D S Y G P K 14 386 G F I L M D S Y G P K K V L D 14 387 F I L M D S Y G P K K V L D G 14 401 G K T I E T S P S L S R M P N 14 410 L S R M P N Q H A C K L G A N 14 418 A C K L G A N I L L E T F K I 14 422 G A N I L L E T F K I H E M I 14 432 I H E M I R Q E I L E Q V L N 14 437 R Q E I L E Q V L N R V V T R 14 459 F L D L L S N I V M Y A P L V 14 460 L D L L S N I V M Y A P L V L 14 463 L S N I V M Y A P L V L Q S C 14 465 N I V M Y A P L V L Q S C S S 14 469. Y A P L V L Q S C S S K V T E 14 488 L S F L P L Q T V Q R L L K A 14 503 V Q P L L K V S M S M R D c L 14 504 Q P L L K V S M S M R D C L I 14 510 S M S M R D C L I L V L R K A 14 515 D C L I L V L R K A M F A N Q 14 TABLE XXV - 83P4B8 HLA- DRB1*0401 (DR4D 4 ) 15 - mers

Pos ■ 1 Seq . ID

2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

550 L G S L S S S Q C s Q S L S V 14

561 S L S V S Q V H V D V H S H Y 14

575 Y N S V A N E T F C L E I M D 14

582 T F c L E I M D S L R R C L S 14

592 R R c L S Q Q A D V R L M L Y 14

602 R L M L Y E G F Y D V L R R N 14

616 N S Q L A N S V M Q T L L S Q 14

620 A N S V M Q T L L S Q L K Q F 14

628 L S Q L K Q F Y E P K P D L L 14

638 K P D L L P P L K L D A C I L 14

639 P D L L P P L K L D A C I L T 14

648 D A C I L T Q G D K I S L Q E 14

649 A C I L T Q G D K I s L Q E P 14

657 K I S L Q E P L D Y L L C c I 14

668 L c C I Q H C L A w Y K N T V 14

711 N R M I K S E L E D F E L D K 14

720 D F E L D K S A D F s Q S T s 14

742 S A F L V M G V C Ξ V L I E Y 14

750 C E V L I E Y N F S I S s F s 14

757 N F S I s S F S K N R F E D I 14

771 I L S L F M C Y K K L S D I L 14

801 S D S L L S M K F V S S L L T 14

814 L T A L F R D s I Q S H Q Ξ s 14

819 R D S I Q S H Q E S L S V L R 14

842 A V N V A L Q K V Q Q L K E T 1.4

847 L Q K V Q Q L K E T G H V S G 14

850 V Q Q L K E T G H V S G P D G 14

867 P E K I F Q N L C D I T R V L 14

874 L c D I T R V L L R Y T S I 14

877 I T R V L L W R Y T S I P T s 14

889 P T S V E E S G K K E K G K s 14

946 D A D V s V T Q R T A F Q I R 14

967 L N L L S S Q E E D F N S K E 14

981 E A L L L V T V L T S L S K L 14

986 V T V L T s L S K L L E P s S 14

992 L S K L L Ξ P S S P Q F V Q M 14

993 S K L L E P s S P Q F V Q M L 14

1003 F V Q M L S w T S K I C K E N 14

1004 V Q M L S W T S K I c K E N S 14

1025 C K S L M N L L F S L H V S Y 14

1028 L M N L L F S L H V S Y K S P 14

1032 L F S L H V S Y K S P V I L L 14

1074 I V N L R T A A P T V C L L V 14

1084 V C L L V L S Q A E K V L E E 14

1085 C L L V L S Q A E K V L E E V 14

1086 L L V L S Q A E K V L E E V D 14

1107 K G Q V S Q E T L s E E A S S 14

1122 Q A T L P N Q P V E K A I I M 14

1131 E K A I I M Q L G T L L T F F 14

1150 Q T A L P S G S C V D T L L K 14

1156 G S C V D T L L K D L C K M Y 14

1159 V D T L L K D L C K M Y T T L 14

1178. R Y Y L Q V C Q S S G G I P K 14

1187 S G G I P K N M E K L V K L S 14

1191 P K N M E K L V K L S G S H L 14

1197 L V K L S G S H L T P L c Y S 14

1205 L T P L C Y S F I S Y V Q N K 14

1240 M A R V L R E T K P I P N L I 14 TABLE XXV - 83P4B8 HLA- DRB1*0401 (DR4D 4 ) 15 - mers

Seq. .^'

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1241 A R V L R E T K P I P N L I F 14 1247 T K P I P N L I F A I E Q Y E 14 1250 I P N L I F A I E Q Y E K F L 14 1254 I F A I E Q Y E K F L I H L S 14 1262 K F L I H L S K K S K V N L M 14 1272 K V N L M Q H M K L s T S R D 14 1273 V N L M Q H M K L S T s R D F 14 1276 M Q H M K L S T S R D F K I K 14 1278 H M K L s T S R D F K I K G N 14 1286 D F K I K G N I L D M V L R E 14 1294 L D M V L R E D G E D E N E E 14

TABLE XXVI - 83P4B8 HLA- DRB1*1101 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 Seq. ID

1 2 3 4 5 score Num.

606 Y E G F Y D V L R R N S Q L A 31

1191 P K N M E K L V K L s G s H L 28

585 L E I M D S L R R C L S Q Q A 27

665 D Y L L C C I Q H C L A w Y K 27

1222 S L N Y T G E K K E K P A A V 27

44 G A L L R A I F K G S P C S E 26

109 A N E F I S A V R E G S L V N 26

906 L L c L E G L Q K I F S A V Q 26

1291 G N I L D M V L R E D G E D E 26

607 E G F Y D V L R R N S Q L A N 25

234 I A F F S A L D K Q H N E E Q 24

537 V A G F L L L L K N F K V L G 24

1170 Y T T L T A L V R Y Y L Q V C 24

174 V I Q H T S M F K D V P L T A 23

189 E E V E F V V E K A L S M F s 23

544 L K N F K V L G S L S S S Q C 23

986 V T V L T S L S K L L E P S s 23

1069 T N H F A I V N L R T A A P T 23

313 L L S V T R I Q R F Q D Q V L 22

335 V K S F K D L Q L L Q G S K F 22

347 S K F L Q N L V P H R S Y V S 22

361 S T M I L E V V K N S V H S W 22

384 E L G F I L M D S Y G P K K V 22

771 I L S L F M C Y K K L S D I L 22

811 s S L L T A L F R D S I Q s H 22

1000 s P Q F V Q M L S T S K I C 22

1177 V R Y Y L Q V C Q s s G G I P 22

442 E Q V L N R V V T R A S S P I 21

443 Q V L N R V V T R A s S P I s 21

490 F L P L Q T V Q R L L K A V Q 21

493 L Q T V Q R L L K A V Q P L L 21

515 D c L I L V L R K A M F A N Q 21

540 F L L L L K N F K V L G S L S 21

566 Q V H V D V H S H Y N S V A N 21

871 F Q N L C D I T R V L L W R Y 21

1039 Y K S P V I L L R D L S Q D I 21

1097 E E V D W L I T K L K G Q V S 21

1100 D L I T K L K G Q V S Q E T 21

1156 G S C V D T L L K D L C K M Y 21

1240 M A R V L R E T K P I P N L I 21

1270 K S K V N L M Q H M K L S T s 21

26 E G D L T N L L Q N Q A V K G 20 TABLE XXVI - 83P4B8 HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

90 I G L L M L E A H H F P G P L 20

91 G L L M L E A H H F P G P L L 20

250 G D E L L D V V T V P S G E L 20

258 T V P S G E L R H V E G T I I 20

310 I A L L L S V T R I Q R F Q D 20

323 Q D Q V L D L L K T S V V K S 20

429 T F K I H E M I R Q E I L E Q 20

460 L D L L S N I V M Y A P L V L 20

500 L K A V Q P L L K V S M S M R 20

514 R D C L I L V L R K A M F A N 20

638 K P D L L P P L K L D A C I L 20

850 V Q Q L K E T G H V S G P D G 20

876 D I T R V L L W R Y T S I P T 20

1007 L S W T s K I C K E N S R E D 20

1081 A P T V c L L V L S Q A E K V 20

1160 D T L L K D L C K M Y T T L T 20

1237 A T A M A R V L R E T K P I P 20

1261 E K F L I H L S K K S K V N L 20

127 L E L L P I I L T A L A T K K 19

214 V Y Q L L V L S S K G S R K S 19

251 D E L L D V V T V P S G E L R 19

778 Y K K L S D I L N E K A G K A 19

882 L W R Y T S I P T S V E E S G 19

982 A L L L V T V L T S L S K L L 19

1030 N L L F S L H V S Y K S P V I 19

1167 C K M Y T T L T A L V R Y Y L 19

1209 C Y S F I S Y V Q N K S K S L 19

17 L Q E F L Q T L R E G D L T N 18

86 V S E I I G L L M L E A H H F 18

211 P P L V Y Q L L V L S S K G S 18

212 P L V Y Q L L V L S S K G S R 18

227 K S V L E G I I A F F S A L D 18

328 D L K T S V V K S F K D L Q 18

407 S P S L s R M P N Q H A C K L 18

466 I V M Y A P L V L Q S C S S K 18

772 L S L F M c Y K K L S D I L N 18

801 S D S L L S M K F V S S L L T 18

954 R T A F Q I R Q F Q R S L L N 18

980 K E A L L L V T V L T S L S K 18

989 L T S L S K L L E P S S P Q F 18

1071 H F A I V N L R T A A P T V C 18

1142 L T F F H E L V Q T A L P s G 18

1176 L V R Y Y L Q V C Q S S G G I 18

1273 V N L M Q H M K L S T S R D F 18

266 H V E G T I I L H I V F A I K 17

283 Y E L G R E L V K H L K V G Q 17

372 V H S W D H V T Q G L V E L G 17

456 I S H F L D L L S N I V M Y A 17

564 V S Q V H V D V H S H Y N S V 17

632 K Q F Y Ξ P K P D L L P P L K 17

696 E A F Y E D L D D I L E S I T 17

741 I S A F L V M G V C E V L I E 17

775 F M C Y K K L S D I L N E K A 17

835 S N E F M R Y A V N V A L Q K 17

1141 L L T F F H E L V Q T A L P S 17

1234 A A V A T A M A R V L R E T K 17

1257 I E Q Y E K F L I H L S K K S 17

1260 Y E K F L I H L S K K S K V N 17 TABLE XXVI - 83P4B8 HLA- DRB1*1101 15 - mers

Seq. ID

Pos • 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

57 S E E A G T L R R R K I Y T C 16

178 T S M F K D V P L T A E E V E 16

199 L S M F S K M N L Q E I P P L 16

365 L E V V K N S V H S W D H V T 16

390 M D S Y G P K K V L D G K T I 16

439 E I L E Q V L N R V V T R A S 16

441 L E Q V L N R V V T R A S S P 16

482 T E A F D Y L S F L P L Q T V 16

487 Y L S F L P L Q T V Q R L L K 16

513 M R D C L I L V L R K A M F A 16

572 H S H Y N s V A N E T F C L E 16

649 A C I L T Q G D K I s L Q E P 16

753 L I E Y N F S I S s F S K N R 16

757 N F S I S S F s K N R F E D I 16

765 K N R F E D I L S L F M C Y K 16

868 E K I F Q N L C D I T R V L L ^' 16

874 L C D I T R V L L w R Y T S I 16

880 V L L R Y T S I P T S V E E 16

913 Q K I F S A V Q Q F Y Q P K I 16

1207 P L C Y S F I S Y V Q N K S K 16

1211 S F I S Y V Q N K S K S L N Y 16

33 Q N Q A V K G K V A G A L L 15

40 G K V A G A L L R A I F K σ S 15

75 L V E S G D L Q K E I V S E I 15

115 A V R Ξ G S L V N G K S L E L 15

131 P I I L T A L A T K K E N L A 15

169 W D Q Q Y V I Q H T s M F K D 15

177 H T S M F K D V P L T A E E V 15

207 Q E I P P L V Y Q L L V L S 15

269 G T I I L H I V F A I K L D Y 15

279 I K L D Y Ξ L G R E L V K H L 15

284 E L G R E L V K H L K V G Q Q 15

331 K T S V V K S F K D L Q L L Q 15

340 D L Q L L Q G S K F L Q N L V 15

348 K F L Q N L V P H R S Y V S T 15

375 W D H V T Q G L V E L G F I L 15

446 N R V V T R A S S P I S H F L 15

450 T R A s S P I s H F L D L L S 15

478 S S K V T E A F D Y L S F L P 15

584 C L E I M D S L R R C L S Q Q 15

677 Y K N T V I P L Q Q G E E E 15

717 E L E D F E L D K s A D F S Q 15

800 T S D S L L S M K F V S S L L 15

890 T S V E E S G K K E K G K S I 15

892 V E E S G K K Ξ K G K S I S L 15

894 Ξ S G K K Ξ K G K S I S L L C 15

924 Q^'P K I Q Q F L R A L D V T D 15

931 L R A L D V T D K E G Ξ E R E 15

946 D A D V S V T Q R T A F Q I R 15

956 A F Q I R Q F Q R S L L N L L 15

983 L L L V T V L T S L S K L L E 15

1032 L F S L H V S Y K S P V I L L 15

1070 N H F A I V N L R T A A P T V 15

1086 L L V L s Q A Ξ K V L E E V D 15

1124 T L P N Q P V E K A I I M Q L 15

1138 L G T L L T F F H E L V Q T A 15

1188 G G I P K N M E K L V K L S •G 15

1213 I S Y V Q N K S K S L N Y T G 15 TABLE XXVI - 83P4B8 HLA- DRBl*noi 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1254 I F A I E Q Y E K F L I H L S 15 1262 K F L I H L S K K S K V N L M 15 1280 K L S T S R D F K I K G N I L 15 1290 K G N I L D M V L R E D G E D 15 http:// v*πv.bι__i-heidelberg.coι_]sy_peithi

TABLE XXIII 109P1D4 HLA-DRB1*0101 15 mers

1 2 Seq. ID

Pos 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

808 S D Y V K I L V A A V A G T I 36

7 T Y I F A V L L A C V V F H s 34

265 G T s V T Q L H A T D A D I G 34

482 G I Q L T K V S A M D A D s G 33

498 N A K I N Y L L G P D A P P E 33

285 H F s F s N L V S N I A R R L 32

173 V Q N Y E L I K S Q N I F G L 31

405 P F R L R P V F S N Q F L L E 30

117 D E I F R L V K I R F L I E D 28

155 N S K Y T L P A A V D P D V G 28

297 R R L F H L N A T T.G L I T I 28

710 E V C Y s I V G G N T R D L F 28

797 N T E I A D V S S P T S D Y V 28

882 L L N F V T I Ξ E T K A D D V 28

945 Q P A F Q I Q P E T P L N S K 28

109 E V E V A I L P D E I F R L V 27

413 S N Q F L L E T A A Y L D Y E 27

807 T S D Y V K I L V A A V A G T 27

90 R I D R E K L C A G I P R D E 26

105 H c F Y Ξ V E V A I L P D E I 26

141 A T V I N I S I P E N S A I N 26

187 L D V I E T P E G D K M P Q L 26

288 F S N L V s N I A R R L F H L 26

430 K E Y A I K L L A A D A G K P 26

431 E Y A I K L L A A D A G K P P 26

538 F T I L A K D N G V P P L T S 26

572 H N E Y N F Y V P E N L P R H 26

596 D P D Y G D N S A V T L S I L 26

738 K C D V T D L σ L H R V L V K 26

823 T V V V V I F I T A V V R c R 26

831 T A V V R C R Q A P H L K A A 26

33 E M P E N V L I G D L L K D L 25

41 G D L L K D L N L S L I P N K 25

62 Q F K L V Y K T G D V P L I R 25

104 E H C F Y E V E V A I L P D E 25

176 Y E L I K S Q N I F G L D V I 25

216 V M K V K V E D G G F P Q R s 25

223 D G G F P Q R S S T A I L Q V 25

296 A R R L F H L N A T T G L I T 25

325 L L V L A S D G G L M P A R A 25

337 A R A M V L V N V T D V N D N 25

433 ^' A I K L L A A D A G K P P L N 25

434 I K L L A A D A G K P P L N Q 25

580 P E N L P R H G T V G L I T V 25

613 N D D F T I D S Q T G V I R P 25 TABLE XXIII 109P1D4 HLA-DRB1* 0101 15 mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

640 T F Y V K A E D G G R V S R S 25 730 T G N I T L M E K C D V T D L 25 764 S V V I V N L F V N E S V T N 25 811 V K I L V A A V A G T I T V V 25 925 P T T F K P D S P D L A R H Y 25 936 A R H Y K S A S P Q P A F Q I 25 27 N Y T I R E E M P E N V L I G 24 46 D L N L S L I P N K S L T T A 24 74 L I R I E E D T G Ξ I F T T G 24 116 P D E I F R L V K I R F L I E 24 145 N I S I P E N S A I N S K Y T 24 322 N H K L L V L A S D G G L M P 24 324 K L L V L A S D G G L M P A R 24 329 A S D G G L M P A R A M V L V 24 331 D G G L M- P A R A M V L V N V 24 . 358 R Y I V N P V N D T V V L S E 24 472 T V S I P E N N S P G I Q L T 24 478 N N S P G I Q L T K V s A M D 24 488 V S A M D A D S G P N A K I N 24 499 A K I N Y L L G P D A P P E F 24 586 H G T V G L I T V T D P D Y G 24 660 T I N V V D V N D N K P V F I 24 670 K P V F I V P P S N C S Y E L 24 698 V I A V D N D T G M N A E V c 24 712 C Y S I V G G N T R D L F A I 24 745 G L H R V L V K A N D L G Q P 24 760 D S L F S V V I V N L F V N E 24 822 I T V V V V I F I T A V V R C 24 885 F V T I E E T K A D D V D S D 24 900 G N R V T L D L P I D L E E Q 24 919 Y N V T T P T T F K P D S P 24 975 C D S I S K C S S S S S D P Y 24 3 L L S G T Y I F A V L L A c V 23

45 K D L N L S L I P N K S L T T 23 78 E E D T G E I F T T G A R I D 23 129 I E D I N D N A P L F P A T V 23 151 N S A I N S K Y T L P ^'A A V D 23 167 D V G I N G V Q N Y E L I K S 23 ^' 281 N A K I H F S F S N L V S N I 23 289 S N L V S N I A R R L F H L N 23 342 L V N V T D V N D N V P S I D 23 349 N D N V P S I D I R Y I V N P 23 370 L S E N I P L N T K I A L I T 23 379 K I A L I T V T D K D A D H N 23 531 E K E D K Y L F T I L A K D N 23 534 D K Y L F T I L A K D N G V P 23 547 V P P L T S N V T V F V S I I 23 630 S F D R E K Q E S Y T F Y V K 23 648 G G R V S R s S S A K V T I N 23 663 V V D V N D N K P V F I V P P 23 669 N K P V F I V P P S N c S Y E 23 679 N C S Y E L V L P S T N P G T 23 680 C S Y E L V L P S T N P G T V 23 782 I N E L V R K S T E A P V T P 23 812 K I L V A A V A G T I T V V V 23 819 A G T I T V V V V I F I T A V 23 TABLE XXIII 109P1D4 HLA-DRB1* 0101 15 mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

821 T I T V V V V I F I T A V V R 23

824 V V V V I F I T A V V R C R Q 23

844 A A Q K N K Q N S E w A T P N 23

916 M G K Y N W V T T P T T F K P 23

963 I Q E L P L D N T F V A C D S 23

6 G T Y I F A V L L A c V V F H 22

126 R F L I E D I N D N A P L F P 22

132 I N D N A P L F P A T V I N I 22

178 L I K S Q N I F G L D V I E T 22

251 Ξ T E I E V S I P E N A P V G 22

328 L A S D G G L M P A R A M V L 22

402 H E I P F R L R P V F S N Q F 22

442 G K P P L N Q S A M L F I K V 22

462 N A P V F T Q S F V T V s I P 22

485 L T K V S A M D A D S G P N A 22

502 N Y L L G P D A P P E F s L D 22

510 P P Ξ F S L D C R T G M L T V 22

535 K Y L F T I L A K D N G V P P 22

544 D N G V P P L T S N V T V F V 22

557 F V S I I D Q N D N S P V F T 22

615 D F T I D S Q T G V I R P N I 22

683 E L V L P S T N P G T V V F Q 22

692 G T V V F Q V I A V D N D T G 22

753 A N D L G Q P D S L F S V V I 22

756 L G Q P D S L F S V V I V N L 22

759 P D S L F S V V I V N L F V N 22

800 I A D V S S P T S D Y V K I L 22

815 V A A V A G T I T V V V V I F 22

939 Y K S A S P Q P A F Q I Q P E 22

947 A F Q I Q P Ξ T P L N s K H H 22

1001 F E V P V S V H T R P V G I Q 22

60 A M Q F K L V Y K T G D V P L 21

108 Y E V E V A I L P D E I F R L 21

184 I F G L D V I E T P E G D K M 21

363 P V N D T V V L s Ξ N I P L N 21

541 L A K D N 'G V P P L T s N V T 21

722 D L F A I D Q E T G N I T L M 21

143 V I N I s I P E N S A I N S K 20

215 Y V M K V K V E D G G F P Q R 20

222 E D G G F P Q R S S T A I L Q 20

246 H P V F K Ξ T E I E V S I P E 20

253 E I E V s I P E N A P V G T S 20

323 H K L L V L A S D G G L M P A 20

346 T D V N D N V P S I D I R Y I 20

425 D Y E S T K E Y A I K L L A A 20

459 E N D N A P V F T Q S F V T V 20

463 A P V F T Q S F V T V S I P E 20

470 F V T V s I P E N N S P G I Q 20

522 L T V V K K L D R E K E D K Y 20

619 D S Q T G V I R P N I S F D R 20

768 V N L F V N E S V T N A T L I 20

783 N E L V R K S T E A P V T P N 20

883 L N F V T I E E T K A D D V D 20

944 P Q P A F Q I Q P E T P L N S 20

992 S D C G Y P V T T F E V P V S 20

63 F K L V Y K T G D V P L I R I 19 TABLE XXIII

109P1D4 HLA-DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

64 K L V Y K T G D V P L I R I E 19

122 L V K I R F L I E D I N D N A 19

182 Q N I F G L D V I E T P E G D 19

306 T G L I T I K E P L D R E E T 19

352 V P S I D I R Y I V N P V N D 19

365 N D T V V L S E N I P L N T K 19

420 T A A Y L D Y E S T K E Y A I 19

500 K I N Y L L G P D A P P E F S 19

604 A V T L S I L D E N D D F T I 19

696 F Q V I A V D N D T G M N A E 19

733 I T L M Ξ K C D V T D L G L H 19

8 Y I F A V L L A C V V F H S G 18

14 L A C V V F H S G A Q E K N Y 18

40 I G D L L K D L N L S L I P N 18

50 S L I P N K S L T T A M Q F K 18

54 N K s L T T A M Q F K L V Y K 18

81 T G E I F T T G A R I D R E K 18

133 N D N A P L F P A T V I N I S 18

136 A P L F P A T V I N I s I P E 18

170 I N G V Q N Y E L I K s Q N I 18

245 N H P V F K E T E I E V S I P 18

257 S I P E N A P V σ T S V T Q L 18

293 S N I A R R L F H L N A T T G 18

319 E T P N H K L L V L A S D G G 18

411 V F s N Q F L L E T A A Y L D 18

423 Y L D Y E S T K E Y A I K L L 18

450 A M L F I K V K D E N D N A P 18

641 F Y V K A E D G G R V S R S S 18

717 G G N T R D L F A I D Q E T G 18

750 L V K A N D L G Q P D S L F S 18

762 L F S V V I V N L F V N E S V 18

765 V V I V N L F V N E S V T N A 18

778 N A T L I N E L V R K S T Ξ A 18

779 A T L I N E L V R K S T E A P 18

870 K K K K K K H S P K N L L L N 18

918 K Y N w V T T P T T F K P D S 18

986 S D P Y S V S D C G Y P V T T 18

993 D C G Y P V T T F E V P V S V 18

995 G Y P V T T F E V P V S V H T 18

TABLE XXIV

109P1D4 HLA- -DRB1*( 3301 (DR17; 1

: L5 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

40 I G D L L K D L N L S L I P N 38

111 E V A I L P D E I F R L V K I 32

900 G N R V T L D L P I D L E E Q 31

36 E N V L I G D L L K D L N L S 30

74 L I R I E E D T G E I F T T G 29

97 C A G I P R D E H C F Y E V E 29

125 I R F L I E D I N D N A P L F 29

502 N Y L L G P D A P P E F S L D 29

893 A D D V D S D G N R V T L D L 28

365 N D T V V L S E N I P L N T K 27 TABLE XXIV 109P1D4 HLA-DRB1*0301 (DR17 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

605 V T L S I L D E N D D F T I D 27

671 P V F I V P P S N C S Y E L V 27

904 T L D L P I D L E E Q T M G K 27

46 D L N L S L I P N K S L T T A 26

54 N K S L T T A M Q F K L V Y K 26

371 S E N I P L N T K I A L I T V 26

525 V K K L D R E K E D K Y L F T 26

613 N D D F T I D s Q T G V I R P 26

626 R P N I s F D R E K Q E S Y T 26

204 Q K E L D R E E K D T Y V M K 25

275 D A D I G E N A K I H F s F s 25

289 S N L V S N I A R R L F H L N 25

401 D H E I P F R L R P V F s N Q 25

510 P P E F S L D C R T G M L T V 25

566 N S P V F T H N E Y N F Y V P 25

662 N V V D V N D N K P V F I V P 25

713 Y S I V G G N T R D L F A I D 25

116 P D E I F R L V K I R F L I E 24

167 D V G I N G V Q N Y E L I K S 24

395 R V T c F T D H Ξ I P F R L R 24

721 R D L F A I D Q E T G N I T L 24

325 L L V L A s D G G L M P A R A 23

628 N I S F D R E K Q E S Y T F Y 23

945 Q P A F Q I Q P E T P L N S K 23

161 P A A V D P D V G I N G V Q N 22

488 V S A M D A D s G P N A K I N 22

925 P T T F K P D s P D L A R H Y 22

970 N T F V A c D s I S K C S S S 22

165 D P D V G I N G V Q N Y E L I 21

323 H K L L V L A s D G G L M P A 21

405 P F R L R P V F S N Q F L L E 21

538 F T I L A K D N G V P P L T S 21

698 V I A V D N D T G M N A Ξ V C 21

759 P D S L F S V V I V N L F V N 21

963 I Q E L P L D N T F V A C D S 21

63 F K L V Y K T G D V P L I R I 20

128 L I E D I N D N A P L F P A T 20

176 Y E L I K S Q N I F G L D V I 20

288 F S N L V s N I A R R L F H L 20

413 S N Q F L L E T A A Y L D Y E 20

434 I K L L A A D A G K P P L N Q 20

580 P E N L P R H G T V G L I T V 20

696 F Q V I A V D N D T G M N A E 20

803 V S S P T s D Y V K I L V A A 20

861 N R Q M I M M K K K K K K K K 20

908 P I D L Ξ E Q T M G K Y N w V 20

928 F K P D S P D L A R H Y K s A 20

104 E H C F Y E V E V A I L P D E 19

109 E V E V A I L P D E I F R L V 19

117 D E I F R L V K I R F L I E D 19

182 Q N I F G L D V I Ξ T P E G D 19

186 G L D V I E T P E G D K M P Q 19

190 I E T P E G D K M P Q L I V Q 19

198 M P Q L I V Q K E L D R E E K 19

238 S V T D T N D N H P V F K E T 19

305 T T G L I T I K E P L D R E E 19 TABLE XXIV 109P1D4 HLA-DRB1*0301 (DR17 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

331 D G G L M P A R A M V L V N V 19 415 Q F L L E T A A Y L D Y E S T 19 421 A A Y L D Y E S T K E Y A I K 19 452 L F I K V K D E N D N A P V F 19 518 R T G M L T V V K K L D R E K 19 519 T G M L T V V K K L D R E K E 19 567 S P V F T H N E Y N F Y V P E 19 588 T V G L I T V T D P D Y G D N 19 682 Y E L V L P S T N P G T V V F 19 712 C Y S I V G G N T R D L F A I 19 730 T G N I T L M E K C D V T D L 19 746 L H R V L V K A N D L G Q P D 19 791 E A P V T P N T Ξ I A D V S S 19 831 T A V V R C R Q A P H L K A A 19 839 A P H L K A A Q K N K Q N S E 19 862 R Q M I M M K K K K K K K K H 19 864 M I M M K K K K K K K K H S P 19

TABLE XXV 109P1D4 HLA-DRB1*0401 (DR4Dw4) 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

173 V Q N Y E L I K S Q N I F G L 28 285 H F S F S N L V S N I A R R L 28 510 P P E F S L D C R T G M L T V 28 613 N D D F T I D S Q T G V I R P 28 916 M G K Y N V T T P T T F K P 28 40 I G D L L K D L N L S L I P N 26 46 D L N L S L I P N K S L T T A 26 54 N K S L T T A M Q F K L V Y K 26 125 I R F L I E D I N D N A P L F 26 167 D V G I N G V Q N Y E L I K S 26 354 S I D I R Y I V N P V N D T V 26 544 D N G V P P L T S N V T V F V 26 555 T V F V S I I D Q N D N S P V 26 765 V V I V N L F V N E S V T N A 26 779 A T L I N E L V R K S T E A P 26 797 N T E I A D V S S P T S D Y V 26 823 T V V V V I F I T A V V R C R 26 827 V I F I T A V V R C R Q A P H 26 893 A D D V D S D G N R V T L D L 26 963 I Q E L P L D N T F V A C D S 26 7 T Y I F A V L L A C V V F H S 22

16 C V V F H S G A Q E K N Y T I 22 104 E H c F. Y E V E V A I L P D E 22 117 D E I F R L V K I R F L I E D 22 124 K I R F L I E D I N D N A P L 22 297 R R L F H L N A T T G L I T I 22 413 S N Q F L L E T A A Y L D Y E 22 467 T Q S F V T V S I P E N N S P 22 628 N I S F D R E K Q E S Y T F Y 22 670 K P V F I V P P S N C S Y E L ^' 22 679 N C S Y E L V L P S T N P G T 22 721 R D L F A I D Q E T G N I T L 22 TABLE XXV

109P1D4t HLA-DRB1* <040_ . (DR4DW4)

15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

768 V N L F V N E S V T N A T L I 22

807 T S D Y V K I L V A A V A G T 22

882 L L N F V T I E E T K A D D V 22

918 K Y N W V T T P T T F K P D S 22

925 P T T F K P D S P D L A R H Y 22

936 A R H Y K s A S P Q P A F Q I 22

969 D N T F V A C D S I S K C S s 22

998 V T T F E V P V S V H T R P V 22

6 G T Y I F A V L L A C V V F H 20

27 N Y T I R E E M P E N V L I G 20

36 E N V L I G D L L K D L N L S 20

37 N V L I G D L L K D L N L S L 20

41 G D L L K D L N L S L I P N K 20

48 N L S L I P N K S L T T A M Q 20

97 C A G I P R D E H C F Y E V E 20

111 E V A I L P D E I F R L V K I 20

112 V A I L P D E I F R L V K I R 20

122 L V K I R F L I E D I N D N A 20

135 N A P L F P A T V I N I S I P 20

140 P A T V I N I S I P E N S A I 20

143 V I N I S I P E N s A I N S K 20

157 K Y T L P A A V D P D V G I N 20

181 S Q N I F G L D V I E T P E G 20

184 I F G L D V I E T P E G D K M 20

231 s T A I L Q V S V T D T N D N 20

232 T A I L Q V s V T D T N D N H 20

234 I L Q V S V T D T N D N H P V 20

245 N H P V F K Ξ T E I E V S I P 20

253 E I E V S I P E N A P V G T s 20

265 G T S V T Q L H A T D A D i G 20

281 N A K I H F S F S N L V S N I 20

289 S N L V S N I A R R L F H L N 20

312 K E P L D R E E T P N H K L L 20

322 N H K L L V L A S D G G L M P 20

323 H K L L V L A S D G G L M P A 20

331 D G G L M P A R A M V L V N V 20

337 A R A M V L V N V T D V N D N 20

338 R A M V L V N V T D V N D N V 20

349 N D N V P S I D I R Y I V N P 20

357 I R Y I V N P V N D T V V L S 20

358 R Y I V N P V N D T V V L S E 20

365 N D T V V L S E N I P L N T K 20

366 D T V V L S E N I P L N T K I 20

377 N T K I A L I T V T D K D A D 20

379 K I A L I T V T D K D A D H N 20

393 N G R V T C F T D H E I P F R 20

405 P F R L R P V F S N Q F L L E 20

421 A A Y L D Y E S T K E Y A I K 20

472 T V S I P E N N s P σ i Q L T 20

482 G I Q L T K V S A M D A D S G 20

488 V s A M D A D S G P N A K I N 20

498. N A K I N Y L L G P D A P P E 20

522 L T V V K K L D R E K E D K Y 20

534 D K Y L F T I L A K D N G V P 20

547 V P P L T s N V T V F V S I I 20

551 T S N V T V F V S I I D Q N D 20

558 V s I I D Q N D N s P V F T H 20 TABLE XXV

109P1D4 HLA-DRB1* ^•0401 1 !DR4DW4)

15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

580 P E N L P R H G T V G L I T V 20

606 T L S I L D E N D D F T I D S 20

640 T F Y V K A E D G G R V s R S 20

648 G G R V S R S S s A K V T I N 20

658 K V T I N V V D V N D N K P V 20

661 I N V V D V N D N K P V F I V 20

682 Y E L V L P S T N P G T V V F 20

692 G T V V F Q V I A V D N D T G 20

695 V F Q V I A V D N D T G M N A 20

696 F Q V I A V D N D T G M N A E 20

698 V I A V D N D T G M N A E V C 20

704 D T G M N A E V C Y S I V G G 20

712 C Y S I V G G N T R D L F A I 20

720 T R D L F A I D Q E T G N I T 20

723 L F A I D Q E T G N I T L M E 20

738 K C D V T D L G L H R V L V K 20

743 D L G L H R V L V K A N D L G 20

747 H R V L V K A N D L G Q P D S 20

753 A N D L G Q P D S L F S V V I 20

759 P D S L F S V V I V N L F V N 20

762 L F S V V I V N L F V N E s V 20

764 S V V I V N L F V N E S V T N 20

767 I V N L F V N Ξ S V T N A T L 20

769 N L F V N Ξ S V T N A T L I N 20

778 N A T L I N E L V R K S T E A 20

800 I A D V S S P T S D Y V K I L 20

808 s D Y V K I L V A A V A G T I 20

810 Y V K I L V A A V A G T I T V 20

811 V K I L V A A V A G T I T V V 20

812 K I L V A A V A G T I T V V V 20

815 V A A V A G T I T V V V V I F 20

819 A G T I T V V V V I F I T A V 20

821 T I T V V V V I F I T A V V R 20

822 I T V V V V I F I T A V V R C 20

839 A P H L K A A Q K N K Q N S E 20

879 K N L L L N F V T I E E T K A 20

880 N L L L N 'F V T I E E T K A D 20

883 L N F V T I E E T K A D D V D 20

900 G N R V T L D L P I D L E E Q 20

904 T L D L P I D L Ξ E Q T M G K 20

906 D L P I D L E E Q T M G K Y N 20

947 A F Q I Q P E T P L N S K H H 20

959 K H H I I Q E L P L D N T F V 20

960 H H I I Q E L P L D N T F' V A 20

975 C D S I S K C s s S S S D P Y 20

995 G Y P V T T F Ξ V P V S V H T 20

12 V L L A C V V F H S G A Q E K 18

13 L L A C V V F H S G A Q E K N 18

19 F H S G A Q E K N Y T I R E E 18

51 L I P N K S L T T A M Q F K L 18

73 P L I R I E E D T G E I F T T 18

78 E E D T G E I F T T G A R I D 18

85 F T T G A.R I D R E K L C A G 18

113 A I L P D E I F R L V K I R F 18

137 P L F P A T V I N I S I P E N 18

144 I N I S I P E N S A I N s K Y 18

148 I P E N S A I N S K Y T L P A 18 TABLE XXV

109P1D4 HLA-.DRB1*0401 ι (DR4DW4)

15 - mers

Pos 1 2 3 4 Seq. ID

5 6 7 8 9 0 1 2 3 4 5 score Num.

196 D K M P Q L I V Q K E L D R E 18

201 L I V Q K E L D R E E K D T Y 18

220 K V E D G G F P Q R S S T A I 18

228 Q R S S T A I L Q V S V T D T 18

258 I P E N A P V G T S V T Q L H 18

262 A P V G T S V T Q L H A T D A 18

282 A K I H F S F S N L V S N I A 18

293 S N I A R R L F H L N A T T G 18

298 R L F H L N A T T G L I T I K 18

309 I T I K E P L D R E Ξ T P N H 18

341 V L V N V T D V N D N V P S I 18

346 T D V N D N V P S r D I R Y I 18

350 D N V P S I D I R Y I V N P V 18

363 P V N D T V V L S E N I P L N 18

370 L S E N I P L N T K I A L I T 18

385 V T D K D A D H N G R V T c F 18

406 F R L R P V F S N Q F L L E T 18

440 D A G K P P L N Q S A M L F I 18

452 L F I K V K D E N D N A P V F 18

460 N D N A P V F T Q S F V T V S 18

464 P V F T Q S F V T V S I P E N 18

487 K V S A M D A D S G P N A K I 18

531 E K E D K Y L F T I L A K D N 18

556 V F V S I I D Q N D N S P V F 18

568 P V F T H N E Y N F Y V P E N 18

577 F Y V P E N L P R H G T V G L 18

595 T D P D Y G D N S A V T L S I 18

598 D Y G D N S A V T L S I L D E 18

609 I L D E N D D F T I D s Q T G 18

618 I D S Q T G V I R P N I S F D 18

625 I R P N I S F D R E K Q E S Y 18

645 A E D G G R V S R S S s A K V 18

659 V T I N V V D V N D N K P V F 18

689 T N P G T V V F Q V I A V D N 18

740 D V T D L G L H R V L V K A N 18

750 L V K A N D L G Q P D S L F S 18

756 L G Q P D S L F S V V I V N L 18

761 S L F S V V I V N L F V N E S 18

770 L F V N E S V T N A T L I N E 18

775 S V T N A T L I N E L V R K S 18

796 P N T E I A D V S S P T S D Y 18

813 I L V A A V A G T I T V V V V 18

833 V V R C R Q A P H L K A A Q K 18

838 Q A P H L K A A Q K N K Q N S 18

854 w A T P N P E N R Q M I M M K 18

876 H S P K N L L L N F V T I E E 18

890 E T K A D D V D S D G N R V T 18

907 L P I D L E E Q T M G K Y N W 18

929 K P D S P D L A R H Y K S A S 18

930 P D S P D L A R H Y K S A S P 18

962 I I Q E L P L D N T F V A C D 18

992 s D C G Y P V T T F E V P V S 18

L001 F E V P V S V H T R P V G I Q 18

223 D G G F P Q R S S T A I L Q V 17

5 S G T Y I F A V L L A c V V F 16

60 A M Q F K L V Y K T G D V P L 16

64 K L V Y K T G D V P L I R I E 16 TABLE XXV 109P1D4 HLA-DRB1*0401 (DR4Dw4 ) 15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

82 G E I F T T G A R I D R E K L 16

105 H C F Y E V E V A I L P D E I 16

136 A P L F P A T V I N I S I P E 16

182 Q N I F G L D V I E T P E σ D 16

246 H P V F K E T E I E V S I P E 16

283 K I H F S F s N L V s N I A R 16

356 D I R Y I V N P V N D T V V L 16

409 R P V F s N Q F L L E T A A Y 16

420 T A A Y L D Y E S T K E Y A I 16

423 Y L D Y E S T K E Y A I K L L 16

450 A M L F I K V K D E N D N A P 16

463 A P V F T Q s F V T V S I P E 16

535 K Y L F T I L A K D N σ V P P 16

554 V T V F V s I I D Q N D N S P 16

572 H N E Y N F Y V P E N L P R H 16

574 E Y N F Y V P E N L P R H G T 16

575 Y N F Y V P E N L P R H G T V 16

596 D P D Y G D N S A V T L S I L 16

639 Y T F Y V K A E D G G R V s R 16

693 T V V F Q V I A V D N D T G M 16

710 E V C Y s I V G G N T R D L F 16

760 D S L F S V V I V N L F V N E 16

826 V V I F I T A V V R C R Q A P 16

945 Q P A F Q I Q P E T P L N S K 16

151 N S A I N s K Y T L P A A V D 15

953 E T P L N s K H H I I Q E L P 15

1 M D L L S G T Y I F A V L L A 14

9 I F A V L L A C V V F H S G A 14

10 F A V L L A C V V F H S G A Q 14

11 A V L L A C V V F H S G A Q E 14

15 A C V V F H S G A Q E K N Y T 14

44 L K D L N L S L I P N K S L T 14

63 F K L V Y K T G D V P L I R I 14

69 T G D V P L I R I E E D T G E 14

71 D V P L I R I E E D T G E I F 14

72 V P L I R I E E D T G E I F T 14

74 L I R I E E D T G E I F T T G 14

88 G A R I D R E K L C A G I P R 14

107 F Y E V E V A I L P D E I F R 14

109 E V E V A I L P D E I F R L V 14

116 P D E I F R L V K I R F L I E 14

119 I F R L V K I R F L I E D I N 14

126 R F L I E D I N D N A P L F P 14

141 A T V I N I S I P E N S A I N 14

145 N I S I P Ξ N s A I N s K Y T 14

161 P A A V D P D V G I N G V Q N 14

170 I N G V Q N Y E L I K S Q N I 14

175 N Y E L I K S Q N I F G L D V 14

176 Y E L I K S Q N I F G L D V I 14

186 G L D V I E T P E G D K M P Q 14

187 L D V I E T P E G D K M P Q L 14

195 G D K M P Q L I V Q K E L D R 14

200 Q L I V Q K E L D R E E K D T 14

204 Q K E L D R E E K D T Y V M K 14

213 D T Y V M K V K V E D G G F P 14

216 V M K V K V E D G G F P Q R S 14

251 E T E I E V S I P E N A P V G 14 TABLE XXV 109P1D4 HLA-DRB1*0401 (DR4DW4 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

255 E V S I P E N A P V G T S V T 14 261 N A P V G T S V T Q L H A T D 14 288 F S N L V S N I A R R L F H L 14 296 A R R L F H L N A T T G L I T 14 299 L F H L N A T T G L I T I K E 14 305 T T G L I T I K Ξ P L D R E Ξ 14 324 K L L V L A S D G G L M P A R 14 325 L L V L A S D G G L M P A R A 14 339 A M V L V N V T D V N D N V P 14 340 M V L V N V T D V N D N V P S 14 342 L V N V T D V N D N V P S I D 14 367 T V V L S E N I P L N T K I A 14 371 S E N I P L N T K I A L I T V 14 415 Q F L L E T A A Y L D Y E s T 14 431 E Y A I K L L A A D A G K P P 14 433 A I K L L A A D A G K P P L N 14 434 I K L L A A D A G K P P L N Q 14 443 K P P L N Q S A M L F I K V K 14 448 Q S A M L F I K V K D E N D N 14 453 F I K V K D E N D N A P V F T 14 462 N A P V F T Q S F V T V S I P 14 468 Q S F V T V S I P E N N S P G 14 470 F V T V S I P E N N S P G I Q 14 480 S P G I Q L T K V S A M D A D 14 502 N Y L L G P D A P P E F S L D 14 518 R T G M L T V V K K L D R E K 14 519 T G M L T V V K K L D R E K E 14 525 V K K L D R E K E D K Y L F T 14 538 F T I L A K D N G V P P L T S 14 553 N V T V F V S I I D Q N D N s 14 586 H G T V G L I T V T D P D Y G 14 588 T V G L I T V T D P D Y G D N 14 591 L I T V T D P D Y G D N S A V 14 602 N S A V T L S I L D E N D D F 14 604 A V T L s I L D E N D D F T I 14 607 L S I L D E N D D F T I D s Q 14 622 T G V I R P N I S F D R E K Q 14 626 R P N I S F D R E K Q Ξ S Y T 14 656 S A K V T I N V V D V N D N K 14 660 T I N V V D V N D N K P V F I 14 663 V V D V N D N K P V F I V P P 14 669 N K P V F I V P P S N C S Y E 14 671 P V F I V P P S N C S Y E L V 14 681 S Y Ξ L V L P S T N P G T V V 14 683 E L V L P S T N P G T V V F Q 14 708 N A E V C Y S I V G G N T R D 14 713 Y S I V G G N T R D L F A I D 14 730 T G N I T L M E K C D V T D L 14 733 I T L M E K C D V T D L G L H 14 741 V T D L G L H R V L V K A N D 14 773 N E S V T N A T L I N E L V R 14 783 N E L V R K S T E A P V T P N 14 824 V V V V I F I T A V V R C R Q 14 830 I T A V V R c R Q A P H L K A 14 861 N R Q M I M M K K K K K K K K 14 885 F V T I E E T K A D D V D S D 14 913 E Q T M G K Y N W V T T P T T 14 TABLE XXV

109P1D4 : HLA-DRB1* ^0401 . ( ;DR4Dw4 )

15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

919 Y N W V T T P T T F K P D S P 14

932 S P D L A R H Y K S A s P Q P 14

970 N T F V A C D S I S K c s s S 14

988 P Y S V S D C G Y P V T T F E 14

1000 T F E V P V s V H T R P V G I 14

1002 E V P V S V H T R P V G I Q V 14

TABLE XXVI

109P1D4 1 HLA-DRB1" '1103 . 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

535 K Y L F T I L A K D N G V P P 32

827 V I F I T A V V R C R Q A P H 26

116 P D E I F R L V K I R F L I E 25

285 H F S F S- N L V S N I A R R L 25

1000 T F E V P V S V H T R P V G I 25

60 A M Q F K L V Y K T G D V P L 24

518 R T G M L T V V K K L D R E K 23

519 T G M L T V V K K L D R E K E 23

882 L L N F V T I E E T K A D D V 23

289 S N L V S N I A R R L F H L N 22

636 Q E S Y T F Y V K A E D G G R 22

730 T G N I T L M Ξ K C D V T D L 22

779 A T L I N E L V R K S T E A P 22

1002 E V P V s V H T R P V G I Q V 22

12 V L L A C V V F H s G A Q E K 21

37 N V L I G D L L K D L N L s L 21

342 L V N V T D V N D N V P S I D 21

522 L T V V K K L D R E K E D K Y 21

808 S D Y V K I L V A A V A G T I 21

861 N R Q M I M M K K K K K K K K 21

11 A V L L A C V V F H S G A Q E 20

82 G E I F T T G A R I D R E K L 20

105 H C F Y E V E V A I L P D E I 20

212 K D T Y V M K V K V E D G G F 20

265 G T S V T Q Ii H A T D A D I G 20

293 S N I A R R L F H L N A T T G 20

479 N S P G I Q L T K V S A M D A 20

482 G I Q L T K V S A M D A D S G 20

645 A E D G G R V S R S S S A K V 20

932 S P D L A R H Y K S A S P Q P 20

972 F V A C D S I S K C S S S S S 20

136 A P L F P A T V I N I S I P E 19

184 I.F G L D V I E T P E G D K M 19

296 A R R L F H L N A T T G L I T 19

322 N H K L L V L A S D G G L M P 19

463 A P V F T Q S F V T V S I P E 19

660 T I N V V D V N D N K P V F I 19

720 T R D L F A I D Q E T G N I T 19

821 T I T V V V V I F I T A V V R 19

7 . T Y I F A V L L A C V V F H S 18

71 D V P L I R I E E D T G E I F 18

126 R F L I E D I N D N A P L F P 18

155 N S K Y T L P A A V D P D V G 18

182 Q N I F G L D V I E T P E G D 18

213 D T Y V M K V K V E D G G F P 18 TABLE XXVI

109P1D4 I HLA-DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

379 K I A L I T V T D K D A D H N 18

431 E Y A I K L L A A D A G K P P 18

485 L T K V S A M D A D S G P N A 18

498 N A K I N Y L L G P D A P P E 18

510 P P E F S L D C R T G M L T V 18

586 H G T V G L I T V T D P D Y G 18

695 V F Q V I A V D N D T G M N A 18

760 D S L F s V V I V N L F V N E 18

764 S V V I V N L F V N E S V T N 18

797 N T Ξ I A D V S S P T S D Y V 18

993 D c G Y P V T T F E V P V S V 18

104 E H C F Y E V E V A I L P D E 17

117 D E I F R L V K I R F L I E D 17

210 E E K D T Y V M K V K V E D G 17

246 H P V F K Ξ T E I E V S I P E 17

380 I A L I T V T D K D A D H N G 17

449 s A M L F I K V K D Ξ N D N A 17

638 s Y T F Y V K A E D G G R V S 17

670 K P V F I V P P s N C S Y E L 17

693 T V V F Q V I A V D N D T G M 17

744 L G L H R V L V K A N D L G Q 17

819 A G T I T V V V V I F I T A V 17

925 P T T F K P D S P D L A R H Y 17

986 S D P Y s V S D C G Y P V T T 17

138 L F P A T V I N I S I P E N S 16

173 V Q N Y E L I K s Q N I F G L 16

399 F T D H E I P F R L R P V F S 16

450 A M L F I K V K D E N D N A P 16

467 T Q S F V T V S I P E N N S P 16

500 K I N Y L L G P D A P P E F s 16

554 V T V F V S I I D Q N D N S P 16

618 I D S Q T G V I R P N I S F D 16

679 N C S Y E L V L P s T N P G T 16

689 T N P G T V V F Q V I A V D N 16

710 E V C Y S I V G G N T R D L F 16

738 K C D V T D L G L H R V L V K 16

768 V N L F V 'N E S V T N A T L I 16

807 T S D Y V K I L V A A V A G T 16

916 M G K Y N V T T P T T F K P 16

936 A R H Y K S A S P Q P A F Q I 16 http://www.b___i-heidelberg.com/sylpeit__i

TABLE XXIII - 151P1C7A - DICKOPF-1 HLA-DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

10 T R V F V A M V A A A L G G H 33 1 M M A L G A A G A T R V F V A 26 203 R H F S K I C K P V L K E G 26 15 A M V A A A L G G H P L L G V 25 14 V A M V A A A L G G H P L L G 24 20 A L G G H P L L G V S A T L N 24 42 I K N L P P P L G G A A G H P 24 71 G N K Y Q T I D N Y Q P Y P C 24 TABLE XXIII - 151P1C7A - DICKOPF-1 HLA-DRB1*0101 15 mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

165 L D G Y S R R T T L S S K M Y 24

11 R V F V A M V A A A L G G H P 23

23 G H P L L G V S A T L N S V L 23

30 S A T L N S V L N S N A I K N 23

39 S N I K N L P P P L G G A A 23

185 E G S V C L R S S D C A S G L 23

24 H P L L G V S A T L N S V L N 22

31 A T L N S V L N s N A I K N L 22

147 R G E I E E T I T E S F G N D 22

129 P G N .Y C K N G I C V S S D Q 21

176 S K M Y H T K G Q E G S V C L 21

143 Q N H F R G Ξ I E E T I T E S 20

232 E I F Q R C Y C G E G L S C R 20

247 I Q K D H H Q A S N S S R L H 20

61 S A A P G I L Y P G G N K Y Q 19

93 T D E Y C A S P T R G G D A G 19

105 D A G V Q I C L A C R K R R K 19

202 R H F s K I C K P V L K E 19

220 C T K H R R K G S H G L E I F 19

7 A G A T R V F V A M V A A A L 18

18 A A A L G G H P L L G V S A T 18

34 N S V L N S N A I K N L P P P 18

55 H P G S A V S A A P G I L Y P 18

63 A P G I L Y P G G N K Y Q T I 18

188 V C L R S S D C A S G L C C A 18

194 D C A S G L C C A R H F W S K 18

244 S C R I Q K D H H Q A S N S S 18

33 L N S V L N S N A I K N L P P 17

43 K N L P P P L G G A A G H P G 17

151 E E T I T E S F G N D H S T L .17

155 T E S F G N D H S T L D G Y S 17

167 G Y S R R T T L S S K M Y H T 17

222 K H R R K G S H G L E I F Q R 17

^'8 G A T R V F V A M V A A A L G 16

9 A T R V F V A M V A A A L G G 16

26 L L G V S A T L N S V L N S N 16 38 N S N A I K N L P P P L G G A 16 45 L P P P L G G A A G H P G S A 16 46 P P P L G G A A G H P G S A V 16 49 L G G A A G H P G S A V S A A 16 51 G A A G H P G S A V S A A P G 16 57 G S A V S A A P G I L Y P G G 16 95 E Y C A S P T R G G D A G V Q 16 99 S P T R G G D A G V Q I C L A 16 128 C P G N Y C K N G I C V S S D 16 159 G N D H S T L D G Y S R R T T 16 168 Y S R R T T L S S K M Y H T K 16 175 S S K M Y H T K G Q E G S V C 16 178 M Y H T K G Q E G S V C L R S 16 196 A S G L c C A R H F w S K I C 16 208 K I C K P V L K E G Q V C T K 16 246 R I Q K D H H Q A S N S S R L 16

TABLE XXIV - 151P1C7A - DICKOPF-1 HLA-DRB1*0301 (DR17) 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Seq. ID Num .

136 G I C V S S D Q N H F R G E I 27

26 L L G V S A T L N S V L N S N 25

63 A P G I L Y P G G N K Y Q T I 23

244 S C R I Q K D H H Q A S N s S 22

34 N S V L N S N A I K N L P P P 20

57 G S A V S A A P G I L Y P G G 20

72 N K Y Q T I D N Y Q P Y P C A 20

210 C K P V L K E G Q V C T K H R 20

10 T R V F V A M V A A A L G G H 19

30 S A T L N S V L N S N A I K N 19

147 R G E I E E T I T E S F G N D 19

155 T E S F G N D H S T L D G Y S 19

188 V C L R S S D C A S G L C C A 19

64 P G I L Y P G G N K Y Q T I D 18

82 P Y P C A E D E Ξ c G T D E Y 18

109 Q I c L A C R K R R K R C M R 18

107 G V Q I C L A C R K R R K R C 17

123 R H A M C C P G N Y C K N G I 17

135 N G I C V S S D Q N H F R G E^' 17

162 H S T L D G Y S R R T T L S S 17

216 Ξ G Q V C T K H R R K G S H G 17

227 G S H G L E I F Q R C Y C G E 17

228 S H G L E I F Q R C Y c G E G 16

240 G E G L S C R I Q K D H H Q A 16

143 Q N H F R σ E I E E T I T E S 15

23 G H P L L G V s A T L N S V L 14

101 T R G G D A G V Q I C L A C R 14

163 s T L D G Y S R R T T L S S K 14

203 R H F W S K I C K P V L K E G 14

11 R V F V A M V A A A L G G H P 13

33 L N S V L N S N A I K N L P P 13

39 S N A I K N L P P P L G G A A 13

42 I K N L P P P L G G A A G H P 13

134 K N G I C V S S D Q N H F R G 13

151 E E T I T E s F G N D H S T L 13

160 N D H s T L D G Y S R R T T L 13

187 S V C L R S S D C A S G L C C 13

TABLE XXV - 151P1C7A -

DICKOPF- 1 HLA-DRB1 *0401

(DR4Dw4 ) 15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

30 S A T L N S V L N S N A I K N 26

196 A S G L C C A R H F W S K I C 26

210 C K P V L K E G Q V C T K H R 26

244 S C R I Q K D H H Q A s N S S 26

71 G N K Y Q T I D N Y Q P Y P C 22

143 Q N H F R G E I E E T I T E S 22

155 T E S F G N D H S T L D G Y S 22

203 R H F W S K I C K P V L K E G 22

1 M M A L G A A G A T R V F V A 20

9 A T R V F V A M V A A A L G G 20

23 G H P L L G V S A T L N S V L 20

24 H P L L G V S A T L N S V L N 20

26 L L G V S A T L N S V L N S N 20

64 P G I L Y P G G N K Y Q T I D 20

107 G V Q I C L A C R K R R K R C 20

123 R H A M C C P G N Y C K N G I 20

136 G I C V S S D Q N H F R G E I 20

162 H S T D G Y S R R T T L S S 20 TABLE XXV - 151P1C7A -

DICKOPF- 1 HLA-DRB1* 0401

(DR4Dw4 ) 15 - mers

Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

S H G L E I F Q R C Y C G E G 20

A L G A A G A T R V F V A M V 18

L G V S A T L N S V L N S N A 18

A T L N S V L N S N A I K N L 18

P P L G G A A G H P G s A V S 18

C K N G I C V S S D Q N H F R 18

E T I T E S F G N D H s T L D 18

Y H T K G Q E G S V C L R S S 18

L D G Y S R R T T L s S K M Y 17

S K M Y H T K G Q E G S V C L 17

T R V F V A M V A A A L G G H 16

G I L Y P G G N K Y Q T I D N 16

Y Q P Y P C A E D E E c G T D 16

T D E Y C A S P T R G G D A G 16

P G N Y C K N G I C V S S D Q 16

Q R C Y C G E G L S e R I Q K 16

E G S V C L R S S D c A S G L 15

R V F V A M V A A A L G G H P 14

F V A M V A A A L G G H P L L 14 y A M V A A A L G G H P L L G 14

A A A L G G H P L L G V S A T 14

L N S V L N S N A I K N L P P 14

N S V L N S N A I K N L P P P 14

S N A I K N L P P P L G G A A 14

I K N L P P P L G G A A G H P 14

P P P L G G A A G H P G S A V 14

G S A V S A A P G I L Y P G G 14

D A G V Q I C L A C R K R R K 14

K R C M R H A M C C P G N Y C 14

R G E I E E T I T E S F G N D 14

K P V L K E G Q V C T K H R R 14

A A G A T R V F V A M V A A A 12

A G A T R V F V A M V A A A L 12

G A T R V F V A M V A A A L G 12

A M V A A A L G G H P L L G V 12

M V A A A L G G H P L L G V S 12

A L G G H P L L G V S A T L N 12

L G G H P L L G V S A T L N S 12

G G H P L L G V s A T L N S V 12

P L L G V S A T L N S V L N S 12

T L N S V L N S N A I K N L P 12

V L N S N A I K N L P P P L G 12

G G A A G H P G S A V S A A P 12

A G H P G S A V S A A P G I L 12

P G S A V S A A P G I L Y P G 12

S A A P G I L Y P σ G N K Y Q 12

Y P G G N K Y Q T I D N Y Q P 12

P Y P C A E D E E c G T D E Y 12

D E E C G T D E Y c A S P T R 12

S P T R G G D A G V Q I C L A 12

T R G G D A G V Q I C L A C R 12

R G G D A G V Q I c L A c R K 12

R K R R K R C M R H A M C C P 12 ^' Y C K N G I C V S s D Q N H F 12

V s S D Q N H F R G E I E E T ^' 12

N H F R G E I E E T I T E S F 12

H F R G E I E E T I T E S F G 12 TABLE XXV - 151P1C7A -

DICKOPF-1 HLA-DRB1* 0401

(DR4Dw4 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

146 F R G E I E E T I T E S F G N 12

149 E I E E T I T E s F G N D H S 12

154 I T E S F G N D H S T L D G Y 12

161 D H S T L D G Y S R R T T L S 12

163 S T L D G Y S R R T T L S S K 12

167 G Y S R R T T L S S K M Y H T 12

168 Y S R R T T L S S K M Y H T K 12

173 T L S S K M Y H T K G Q E G s 12

182 K G Q E G S V C L R S S D C A 12

184 Q E G S V C L R S S D c A S G 12

188 V C L R S S D C A S G L C C A 12

199 L C C A R H F S K I C K P V 12

207 s K I C K P V L K E G Q V C T 12

208 K I C K P V L K E G Q V C T K 12

213 V L K E G Q V C T K H R R K G 12

215 K E G Q V C T K H R R K G S H 12

217 G Q V C T K H R R K G S H G L 12

227 G S H G L E I F Q R C Y ^■C G E 12

236 R C Y C G E G L S C R I Q K D 12

237 C Y c G E G L S C R I Q K D H 12

238 Y c G E G L S C R I Q K D H H 12

245 c R I Q K D H H Q A S N S S R 12

247 I Q K D H H Q A S N S S R L H 12

248 Q K D H H Q A S N S S R L H T 12

249 K D H H Q A S N S S R L H T C 12

251 H H Q A S N S S R L H T C Q R 12

TABLE XXVI - 151P1C7A - DICKOPF-1 HLA-DRB1*1101 15 mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

203 R H F W S K I C K P V L K E G 25

71 G N K Y Q T I D N Y Q P Y P C 22

216 E G Q V C T K H R R K G S H G 22

172 T T L S S K M Y H T K G Q E G 21

196 A S G L c c A R H F w S K I C 20

244 S C R I Q K D H H Q A S N s S 20

11 R V F V A M V A A A L G G H P 19

39 S N A I K N L P P P L G G A A 19

10 T R V F V A M V A A A L G G H 18

30 S A T L N S V L N S N A I K N 18

65 G I L Y P G G N K Y Q T I D N 18

93 T D E Y C A S P T R G G D A G 17

129 P G N Y C K N G I C V S S D Q 17

107 G V Q I C L A C R K R R K R C 16

109 Q I C L A C R K R R K R C M R 16

137 I C V S S D Q N H F R G E I E 16

165 L D G Y S R R T T L S S K M Y 16

176 S K M Y H T K G Q E G S V C L 16

217 G Q V C T K H R R K G S H G L 16

231 L E I F Q R C Y C G E G L S C 16

35 S V L N S N A I K N L P P P L 15

162 H S T L D G Y s R R T T L S S 15

9 A T R V F V A M V A A A L G G 14

47 P P L G G A A G H P G S A V S 14 116 K R R K R C M R H A M C C P G 14 TABLE XXVI - 151P1C7A - DICKOPF- 1 HLA-DRB1*1101 15 mers

1 2 3 4 5 6 7 Seq. ID

Pos 8 9 0 1 2 3 4 5 score Num.

206 W S K I C K P V L K E G Q V C 14 207 S K I c K P V L K E G Q V C T 14 228 S H G L E I F Q R C Y C G E G 14 241 E G L s C R I Q K D H H Q A S 14 243 L S C R I Q K D H H Q A S N S 14 23 G H P L L G V S A T L N S V L 13 24 H P L L G V S A T L N S V L N 13 102 R G G D A G V Q I C L A c R K 13 182 K G Q E G S V C L R S S D C A 13 8 G A T R V F V A M V A A A L G 12

14 V A M V A A A L G G H P L L G 12

20 A L G G H P L L G V S A T L N 12

36 V L N S N A I K N L P P P L G 12

42 I K N L P P P L G G A A G H P 12

43 K N L P P P L G G A A G H P G 12

46 P P P L G G A A G H P G S A V 12

54 G H P G S A V S A A P G I L Y 12

60 V S A A P G I L Y P G G N K Y 12

119 K R C M R H A M C C P G N Y c 12

148 G E I Ξ E T I T E S F G N D H 12

175 S S K M Y H T K G Q E G S V C 12

185 E G s V C L R S S D C A S G L 12

187 S V c L R S S D C A S G L C C 12 http://www.brni-heidelberg.com/syfpeithi

TABLE XXIII - 151P4E11 - HLA- DRB1*0101 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

105 L D R L L D L P A A A S S E D 32 31 K R G W T L N S A G Y L L G P . 28 93 F L S F L H L K E A G A L D R 28 51 H R S F S D K N G L T S K R E 26 91 I E F L S ,F L H L K E A G A L 26 94 L S F L H L K Ξ A G A L D R L 26

2 A R G S A L L L A S L L L A A 25

7 L L L A S L L L A A A L S A S 25

8 L L A S L L L A A A L S A S A 25

9 L A S L L L A A A L S A S A G 25

88 R T I I E F L S F L H L K E A 25 106 D R L L D L P A A A S S Ξ D I 25

6 A L L L A S L L L A A A L S A 24

15 A A A L S A S A G L W S P A K 24 36 L N S A G Y L L G P H A V G N 24 37 N S A G Y L L G P H A V G N H 24 84 N N I M R T I I E F L S F L H 24 102 A G A L D R L L D L P A A A S 24

3 R G S A L L L A S L L L A A A 23

11 S L L L A A A L S A S A G L W 22 66 L R P E D D M K P G S F D R S 20 63 K R Ξ L R^' P E D D M K P G S F 19 85 N I M R T I I E F L S F L H L 19 103 G A L -D R L L D L P A A A S S 19 39 A G Y L L G P H A V G N H R S 18 96 F L H L K E A G A L D R L L D 18 TABLE XXIII - 151P4E11 - HLA- DRB1* 01-01 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

4 G S A L L L A S L L L A A A L 17

13 L L A A A L S A S A G L W S P 17 25 W S P A K E K R G W T L N S A 17 57 K N G L T S K R E L R P E D D 17 69 E D D M K P G S F D R s I P E 17

1 M A R G S A L L L A S L L L A 16

5 S A L L L A S L L L A A A L S 16

10 A S L L L A A A L S A s A G L 16 12 L L L A A A L S A S A G L W S 16 30 E K R G W T L N S A G Y L L G 16 32 R G T L N S A G Y L L G P H 16 54 F S D K N G L T S K R E L R P 16 60 L T S K R E L R P E D D M K P 16 74 P G S F D R S I P E N N I M R 16 75 G S F D R S I P E N N I M R T 16 80 S I P E N N I M R T I I E F L 16 81 I P E N N I M R T I I E F L S 16 83 E N N I M R T I I E F L S F L 16 27 P A K E K R G w T L N S A G Y 15 42 L L G P H A V G N H R S F S D 15 104 A L D R L L D L P A A A S S E 15

TABLE XXIV - 151P4E11 HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

94 L S F L H L K E A G A L D R L 21 88 R T I I E F L S F L H L K E A 20 100 K E A G A L D R L L D L P A A 20 50 N H R S F S D K N G L T S K R 19 57 K N G L T S K R E L R P E D D 19 21 S A G L W s P A K E K R G W T 18 83 E N N I M R T I I E F L S F L 18 84 N N I M R T I I E F L S F L H 18 93 F L s F L H L K E A G A L D R 18

4 G S A L L L A s L L L A A A L 16

55 S D K N G L T s K R Ξ L R P E 16 65 E L R P E D D M K P G S F D R 16 104 A L D R L L D L P A A A S s E 16

5 S A L L L A S L L L A A A L S 15

49 G N H R S F S D K N G L T S K 15 105 L D R L L D L P A A A S S E D 15

6 A L L L A S L L L A A A L S A 14

9 L A S L L L A A A L S A S A G 14

33 G W T L N S A G Y L L G P H A 14 87 M R T I I E F L S F L H L K E 14 90 I I E F L S F L H L K E A G A 14 96 F L H L K E A G A L D R L L D 14

3 R G S A L L L A S L L L A A A 13

10 A S L L L A A A L S A S A G L 13 39 A G Y L L G P H A V G N H R S 13 63 K R E L R P E D D M K P G S F 13 69 E D D M K P G S F D R s I P E 13

7 L L L A S L L L A A A L s A S 12

11 S L L L A A A L S A S A G L w 12 40 G Y L L G P H A V G N H R S F 12 102 A G A L D R L L D L P A A A S 12 106 D R L L D L P A A A S S E D I 12 TABLE XXIV - 151P4E11 HLA- DRB1* 0301 (DR17 ) 15 - mers

POS 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Seq. ID Num.

2 A R G S A L L L A S L L L A A 11

15 A A A L S A S A G L W S P A K 11 31 K R G W T L N S A G Y L L G P 11 32 R G W T L N S A G Y L L G P H 11 64 R E L R P E D D M K P G S F D 11 75 G S F D R S I P E N N I M R T 11 78 D R S I P E N N I M R T I I Ξ 11 85 N I M R T I I E F L S F L H L 11

1 M A R G S A L L L A S L L L A 10

13 L L A A A L S A S A G L W S P 10 25 W S P A K E K R G W T L N S A 10 45 P H A V G N H R S F S D K N G 10 56 D K N G L T S K R E L R P E D 10 72 M K P G S F D R S I P E N N I 10 82 P E N N I M R T I I Ξ F L S F 10 91 I E F L S F L H L K Ξ A G A L^' 10 98 H L K E A G A L D R L L D L P 10 108 L L D L P A A A S S E D I E R 10

TABLE XXV - 151P4E11 HLA- DRB1*0401 (DR4DW4 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

11 S L L L A A A L S A S A G L W 26 90 I I E F L S F L H L K E A G A 22 93 F L S F L H L K E A G A L D R 22

4 G S A L L L A S L L L A A A L 20

6 A L L L A S L L L A A A L S A 20

9 L A S L L L A A A L S A S A G 20

10 A S L L L A A A L S A S A G L 20

84 N N I M R T I I E F L S F L H 20

87 M R T I I E F L S F L H L K Ξ 20

88 R T I I E F L S F L H L K E A 20

91 I E F L S F L H L K E A G A L 20

102 A G A L D R L L D L P A A A S 20

105 L D R L L D L P A A A S S E D 20

108 L L D L P A A A S S E D I E R 20

3 R G S A L L L A S L L L A A A 18

18 L S A S A G L W S P A K E K R 18 24 L W S P A K E K R G T L N S 18 42 L L G P H A V G N H R s F S D 18 50 N H R S F S D K N G L T S K R 18 80 S I P E N N I M R .T I I E F L 18 109 L D L P A A A S S E D I E R S 18 22 A G L W S P A K E K R G W T L 16 31 K R G W T L N S A G Y L L G P 16 38 S^' A G Y L L G P H A V G N H R 16 74 P G S F D R S I P E N N I M R 16

5 S A L L L A S L L L A A A L S 14

15 A A A L S A S A G L W S P A K 14 33 G W T L N S A G Y L L G P H A 14 39 A G Y L L G P H A V G N H R S 14 45 P H A V G N H R S F S D K N G 14 ^' 63 K R Ξ L R P E D D M K P G S F 14 69 Ξ D D M K P G S F D R S I P E 14 78 D R S I P E N N I M R T I I E 14 83 E N N I M R T I I E F L s F. L 14 96 F L H L K E A G A L D R L L D 14 TABLE XXV - 151P4E11 HLA- DRB1* 0401 (DR4DW4 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

106 D R L L D L P A A A S S E D I 14

1 M A R G S A L L L A s L L L A 12

2 A R G S A L L L A S L L L A A 12

7 L L L A S L L L A A A L S A S 12

8 L L A S L L L A A A L S A S A 12

13 L L A A A L S A S A G L W S P 12 27 P A K E K R G T L N S A G Y 12 30 E K R G W T L N s A G Y L L G 12 36 L N S A G Y L L G P H A V G N 12 37 N S A G Y L L G P H A V G N H 12 44 G P H A V G N H R S F S D K N 12 47 A V G N H R S F S D K N G L T 12 54 F S D K N G L T S K R E L R P 12 59 G L T S K R E L R P E D D M K 12 65 E L R P Ξ D D M K P G S F D R 12 71 D M K P G S F D R s I P Ξ N N 12 72 M K P G S F D R S I P E N N I 12 75 G S F D R S I P E N N I M R T 12 76 S F D R S I P E N N I M R T I 12 77 F D R S I P E N N I M R T I I 12 86 I M R T I I E F L s F L H L K 12 98 H L K E A G A L D R L L D L P 12 99 L K E A G A L D R L L D L P A 12 100 K E A G A L D R L L D L P A A 12 103 G A L D R L L D L P A A A S S 12 104 A L D R L L D L P A A A S S E 12

TABLE XXVI - 151P4E11 HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

38 S A G Y L L G P H A V G N H R 25 93 F L S F L H L K E A G A L D R 22 84 N N I M R T I I E F L S F L H 19 90 I I E F L S F L H L K E A G A 19

6 A L L L A S L L L A A A L S A 18

22 A G L W S P A K E K R G W T L 18 91 I E F L S F L H L K E A G A L 18 102 A G A L D R L L D L P A A A S 18 105 L D R L L D L P A A A S S E D 18 51 H R S F S D K N G L T S K R E 16 57 K N G L T S K R E L R P E D D 16 21 S A G L S P A K E K R G T 15 71 D M K P G S F D R S I P E N N 15 80 S I P E N N I M R T I I E F L 15 92 E F L S F L H L K E A G A L D 15 99 L K E A G A L D R L L D L P A 15 42 L L G P H A V G N H R S F S D 14 44 G P H A V G N H R S F S D K N 14 65 E L R P E D D M K P G S F D R 14 69 E D D M K P G S F D R S I P E 14

4 G S A L L L A S L L L A A A L 13

8 L L A S L L L A A A L S A S A 13

33 G W T L N S A G Y L L G P H A 13 63 K R E L R P E D D M K P G S F 13

5 S A L L L A S L L L A A A L S 12

9 L A S L L L A A A L S A S A G 12 10 A S L L L A A A L S A S A G L 12 TABLE XXVI - 151P4E11 HLA- DRB1*1101 15 - mers

Pos 1 2 3 4 8 9 0 1 2 Seq. ID

5 6 7 3 4 5 score Num.

11 S L L L A A A L S A S A G L W 12 12 L L L A A A L S A S A G L W S 12 15 A A A L S A S A G L W S P A K 12 31 K R G W T L N S A G Y L L G P 12 36 L N S A G Y L L G P H A V G N 12 88 R T I I E F L S F L H L K E A 12 103 G A L D R L L D L P A A A S S 12 106 D R L L D L P A A A S S E D I 12

TABLE XXIII - 154P2A8 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

216 V A V L V I L I G C Y I A I S 36

154 S I T F T K V L S V C V W V I 33

251 N Q s I R V V V A V F F T c F 32

93 T F P F R I V H D A G F G P W 31

119 V L F Y A N M Y T S I V F L G 30

162 S V C V V I M A V L S L P N 30

12 N N Ξ L H G Q E S H N S G N R 29

48 L Y L I I F V A S I L L N G L 29

204 H T A V T Y V N S c L F V A V 29

150 S R M Y S I T F T K V L S V C 28

163 V C V w V I M A V L S L P N I 28

46 P V L Y L I I F V A S I L L N 27

47 V L Y L I I F V A S I L L N G 27

95 P F R I V H D A G F G P Y F 27

123 A N M Y T S I V F L G L I S I 27

126 Y T s I V F L G L I S I D R Y 27

4 N L T L A K L P N N E L H G Q 26

78 F Y L K N I V V A D L I M T L 26

9 K L P N N E L H G Q E S H N S 25

53 F V A S I L L N G L A V W I F 25

56 S I L L N G L A V W I F F H I 25

64 V^'W I F F H I R N K T S F I F 25

212 S C L F V A V L V I L •I G c Y 25

229 I s R Y I H K S S R Q F I S Q 25

259 A V F F T C F L P Y H L c R I 25

280 •L D R L L D E S A Q K I L Y Y 25

37 H N E F D T I V L P V L Y L I 24

42 T I V L P V L Y L I I F V A s 24

44 V L P V L Y L I I F V A S I L 24

51 I I F V A S I L L N G L A V W 24

55 A S I L L N G L A V w I F F H 24

67 F F H I R N K T S F I F Y L K 24

74 T S F I F Y L K N I V V A D L 24

87 D L I M T L T F P F R I V H D 24

128 S I V F L G L I S I D R Y L K 24

149 D S R M Y S I T F T K V L S V 24

166 V I M A V L S L P N I I L T 24

168 ^' I M A V L S L P N I I L T N G 24

217 A V L V I L I G C Y I A I S R 24

237 S R Q F I S Q S S R K R K H N 24

254 I R V V V A V F F T c F L P Y 24

277 F S H L D R L L D E s A Q K I 24

45 L P V L Y L I I F V A S I L L 23 t tO W tO t H co cn to o o vo H _o σ_. VD -J θo 0 a in -J co 01

o rt Ki Ki < a a B a Ω a a a a a a a co a Ω X a 3 < a H <; a < CO H ω S a α rt O a H a a < a a H a a a a s α a O < H

H a Ω H <; rt a cn o a < 3 Ω H a ^ a rt a H cn a cn pi a O α a < a a co < Si a a rt a H H Si a Ω a < <: rt a a H rt Ki a a to a a a Ki Ω a a a -3 a H rt rt Ω « Ω a Ω a co a < a rt < a a H a a Ki Ω rt H CO < a a < Ω a a < < a H a a H a Ki D IO α a a H CO <. a H H < rt Ki < < a a a H Ω a cn a H a < Ki o H H K a <! α a a a a a a a Ki a H α H a < a H a < a a a a a rf_* a a a cn Ω a a Ω a H H cn rt > < a cn a a Ki a a a rt Ω Pi a a a < H < a a a Ω a H co rt 3 cn co H o > a Ω a a a Ω a H a H

*

B a a CO s; < o a 3 H cn D a a a a H cn a H a o a a H a H a H a H a a a a a H Ω a > a H a a cn a Ω H a a < O

2 a cn H

CO a Ki a Ω a i Ω i > < < a o a a cn TJ a H rt to a 3 a Ω co a a a a a H -3 a a a a CO <! 2 O Ω H a < Ki a a < H <: rt H O

H a H a rt -3 cn a < a a H a a α a CO H a co a a a Ω <! < Ω Ω rt a a 3 Ω a Ki H a a a a a a Ki a a 2 a Ω rt a a a a a < CO H

Ω a > a a a Ω a a a 3 H o a a a CO Ω a rt < 3 α a a a Ki a a Ω <! a Ω a H a a H a co CO i Ki rt rt < CO rt a in

Ki a a rt VD a a a < < < rt a cn Ω Ki rt 3 rt a Ω KD a < a a a Ki cn a rt Ki H a Ω O rt a a X a Ω rt α a H a rt a Ω Si < o rt a rt a a 1 o

H H a a rt > a a a a a rt H a a a a Ω Ω c cn a a a a < D cn a O rt a a < H a a a a rt a cn rt a < a < cn H a Ω a α 3

H π> a a n <: <: i < CO rt a H a rt a <: α H H H S3 <: H 3 Ω a H H > a a a H O rt H > > < a a a a < a a H a a a H cn O a a H. to cn

Ki a a H a < rt a a a cn 3 a rt a B a co a a a a Ω rt Ki α a H cn < rt 3 a < CO a Ω o < a a H rt a a a 2 Ki a H a a cn a H w

K rt o a <. a K IT α H H a a a cn a KD a rt S3 rt < a CO CO a a a a a * Ω a a < Ω a a > si CO D H a « a a κ; .t_. ; κ; a <: a a H 2 rf_*

Ω a a H H cn Ω a H D rt a a rt O en > a a Si rt H a H < Ki <1 α cn α a a a a Ω H a H a a < a Ki a a < a α 2 < 1^ in

01

H H H H H H H H H H H H H H H H to t to to t to to to to to to t t to to to to O

-o. 1 -J -0 1 -o. vo vo o o o o o o H to to to to to ω ιo ω IO o

TABLE XXIV - 154P2A8 HLA- DRB1* 0301 (DR17 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

81 K N I V V A D L I M T L T F P 30 168 I M A V L S L P N I I L T N G 28

4 N L T L A K L P N N E L H G Q 26

281 D L D E S A Q K I L Y Y C 25

289 Q K I L Y Y C K E I T L F L s 25

65 W I F F H I R N K T S F I F Y 22

132 L G L I S I D R Y L K V V K P 22

51 I I F V A S I L L N G L A V w 21

85 V A D L I M T L T F P F R I V 21

95 P F R I V H D A G F G P Y F 21

215 F V A V L V I L I G C Y I A I 21

48 L Y L I I F V A s I L L N G L 20

77 I F Y L K N I V V A D L I M T 20

109 F K F I L C R Y T S V L F Y A 20

110 K F I L C R Y T s V L F Y A N 20

129 I V F L G L I s I D R Y L K V 20

198 P L G V K H T A V T Y V N S 20

204 H T A V T Y V N s C L F V A V 20

251 N Q S I R V V V A V F F T C F 20

47 V L Y L I I F V A S I L L N G 19

67 F F H I R N K T S F I F Y L K 19

131 F L G L I S I D R Y L K V V K 19

175 P N I I L T N G Q P T E D N I 19

186 E D N I H D C S K L K S P L G 19

211 N S C L F V A V L V I L I G C 19

254 I R V V V A V F F T C F L P Y 19

257 V V A V F F T C F L P Y H L C 19

42 T I V L P V L Y L I I F V A S 18

56 S I L L N G L A V I F F H I 18

63 A V W I F F H I R N K T S F I 18

93 T F P F R I V H D A G F G P 18

116 Y T S V L F Y A N M Y T S I V 18

134 L I S I D R Y L K V V K P F G 18

230 S R Y I H K S S R Q F I S Q S 18

255 R V V V A V F F T C F L P Y H 18

259 A V F F T C F L P Y H L c R I 18

288 A Q K I L Y Y C K E I T L F L 18

73 K T S F I F Y L K N I V V A D 17

74 T S F I F Y L K N I V V A D L 17

89 I M T L T F P F R I V H D A G 17

143 V V K P F G D S R M Y S I T F 17

238 R Q F I S Q S s R K R K H N Q 17

272 R I P F T F S H L D R L L D Ξ 17

25 N R S D G P G K N T T L H N E 16

37 H N E F D T I V L P V L Y L I 16

188 N I H D C S K L K s P L G V K 16

274 P F T F S H L D R L L D E S A 16

282 R L L D E S A Q K I L Y Y C K 16

41 D T I V L P V L Y L I I F V A 15

54 V A S I L L N G L A V w I F F 15

192 C S K L K S P L G V K w H T A 15

203 H T A V T Y V N S c L F V A 15

279 H L D R L L D E S A Q K I L Y 15

280 L D R L L D E S A Q K I L Y Y 15

2 G F N L T L A K L P N N E L H 14

22 N S G N R S D G P G K N T T L 14 55 A S I L L N G L A V W I F F H 14 TABLE XXIV - 154P2A8 HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

64 V I F F H I R N K T S F I F 14 102 A G F G P Y F K F I L C R Y 14 207 V T Y V N S C L F V A V L V I 14 264 C F L P Y H L C R I P F T F S 14

TABLE XXV - 154P2A8 HLA- DRB1*0401 (DR4DW4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

64 V w I F F H I R N K T s F I F 28

154 S I T F T K V L S V c V W V I 28

200 G V K H T A V T Y V N S c L 28

4 N L T L A K L P N N E L H G Q 26

48 L Y L I I F V A S I L L N G L 26

61 G L A V w I F F H I R N K T S 26

74 T S F I F Y L K N I V V A D L 26

131 F L G L I s I D R Y L K V V K 26

165 V W V I M A V L S L P N I I L 26

168 I M A V L S L P N I I L T N G 26

255 R V V V A V F F T c F L P Y H 26

37 H N E F D T I V L P V L Y L I 22

46 P V L Y L I I F V A S I L L N 22

62 L A V w I F F H I R N K T S F 22

106 P W Y F K F I L c R Y T s V L 22

113 L C R Y T S V L F Y A N M Y T 22

119 V L F Y A N M Y T S I V F L G 22

123 A N M Y T S I V F L G L I S I 22

128 S I V F L G L I S I D R Y L K 22

150 S R M Y S I T F T K V L s V c 22

212 S C L F V A V L V I L I G C Y 22

223 I G C Y I A I S R Y I H K S s 22

259 A V F F T C F L P Y H L c R I 22

274 P F T F s H L D R L L D E S A 22

2 G F N L T L A K L P N N Ξ L H 20

12 N N E L H G Q E S H N S G N R 20

33 N T T L H N E F D T I V L P V 20

41 D T I V L P V L Y L I I F V A 20

44 V L P V L Y L I I F V A s I L 20

45 L P V L Y L I I F V A S I L L 20

47 V L Y L I I F V A S I L L N G 20

49 Y L I I F V A s I L L N G L A 20

51 I I F V A s I L L N G L A V W 20

63 A V W I F F H I R N K T S F I 20

77 I F Y L K N I V V A D L I M T 20

80 L K N I V V A D L I M T L T F 20

81 K N I V V A D L I M T L T F P 20

82 N I V V A D L I M T L T F P F 20

116 Y T s V L F Y A N M Y T S I V 20

117 T S V L F Y A N M Y T S I V F 20

126 Y T s I V F L G L I S I D R Y 20

132 L G L I S I D R Y L K V V K P 20

141 L K V V K P F G D S R M Y S I 20

149 D S R M Y s I T F T K V L S V 20

157 F T K V L S V c V w V I M A V 20

162 S V C V W V I M A V L S L P N 20

166 W V I M A V L S L P N I I L T 20

204 H T A V T Y V N S c L F V A V 20

211 N S C L F V A V L V I L I G c 20 TABLE XXV - 154P2A8 HLA- DRB1* 0401 (DR4Dw4 ) 15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

216 V A V L V I L I G C Y I A I S 20

217 A V L V I L I G C Y I A I S R 20

226 Y I A I S R Y I H K s S R Q F 20

230 s R Y I H K S s R Q F I S Q S 20

238 R Q F I S Q S s R K R K H N Q 20

267 P Y H L C R I P F T F s H L D 20

277 F s H L D R L L D E S A Q K I 20

280 L D R L L D E s A Q K I L Y Y 20

281 D R L L D Ξ S A Q K I L Y Y c 20

34 T T L H N Ξ F D T I V L P V L 18

52 I F V A S I L L N G L A V I 18

107 W Y F K F I L C R Y T s V L F 18

143 V V K P F G D S R M Y s I T F 18

172 L S L P N I I L T N G Q P T E 18

185 T E D N I H D C S K L K s P L 18

222 L I G C Y I A I S R Y I H K S 18

234 H K S S R Q F I S Q S s R K R 18

279 H L D R L L D E S A Q K I L Y 18

73 K T S F I F Y L K N I V V A D 16

93 T F P F R I V H D A G F G P W 16

101 D A G F G P w Y F K F I L C R 16

105 G P W Y F K F I L C R Y T S V 16

108 Y F K F I L c R Y T S V L F Y 16

118 s V L F Y A N M Y T S I V F L 16

137 I D R Y L K V V K P F G D S R 16

144 V K P F G D S R M Y S I T F T 16

163 V C V W V I M A V L S L P N I 16

206 A V T Y V N S C L F V A V L V 16

237 s R Q F I s Q S S R K R K H N 16

262 F T C F L P Y H L C R I P F T 16

265 F L P Y H L c R I P F T F S H 16

272 R I P F T F s H L D R L L D E 16

109 F K F I L c R Y T S V L F Y A 15

196 K s P L G V K W H T A V T Y V 15

7 L A K L P N N E L H G Q E S H 14

40 F D T I V L P V L Y L I I F V 14

42 T I V L P V L Y L I I F V A S 14

55 A s I L L N G L A V W I F F H 14

56 s I L L N G L A V W I F F H I 14

59 L N G L A V I F F H I R N K 14

85 V A D L I M T L T F P F R I V 14

86 A D L I M T L T F P F R I V H 14

87 D L I M T L T F P F R I V H D 14

89 I M T L T F P F R I V H D A G 14

95 P F R I V H D A G F G P W Y F 14

96 F R I V H D A G F G P W Y F K 14

110 K F I L c R Y T S V L F Y A N 14

129 I V F L G L I S I D R Y L K V 14

134 L I S I D R Y L K V V K P F G 14

138 D R Y L K V V K P F G D S R M 14

152 M Y s I T F T K V L S V C V W 14

158 T K V L S V C V w V I M A V L 14

160 V L s V c V W V I M A V L S L 14

164 c V w V I M A V L S L P N I I 14

169 M A V L s L P N I I L T N G Q 14

171 V L s L P N I I L T N G Q P T 14

174 L P N I I L T N G Q P T E D N 14

175 P N I I L T N G Q P T E D N I 14 TABLE XXV - 154P2A8 HLA- DRB1*0401 (DR4DW4 ) 15 - mers

Pos ^■ 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Seq. ID Num.

186 E D N I H D C S K L K S P L G 14

207 V T Y V N S C L F V A V L V I 14

213 C L F V A V L V I L I G C Y I 14

215 F V A V L V I L I G c Y I A I 14

218 V L V I L I G C Y I A I S R Y 14

219 L V I L I G C Y I A I S R Y I 14

224 G C Y I A I S R Y I H K S S R 14

251 N Q S I R V V V A V F F T C F 14

253 S I R V V V A V F F T C F L P 14

254 I R V V V A V F F T C F L P Y 14

270 L C R I P F T F S H L D R L L 14

288 A Q K I L Y Y C K E I T L F L 14

289 Q K I L Y Y c K E I T L F L S 14

TABLE XXVI - 154P2A8 HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

137 I D R Y L K V V K P F G D S R 32

223 I G C Y I A I S R Y I H K S S 25

274 P F T F S H L D R L L D E s A 25

64 V W I F F H I R N K T S F I F 23

92 L T F P F R I V H D A G F G P 23

213 C L F V A V L V I L I G C Y I 22

107 Y F K F I L C R Y T S V L F 21

131 F L G L I s I D R Y L K V V K 21

186 E D N I H D c S K L K S P L G 21

123 A N M Y T S I V F L G L I s I 20

162 S V C V V I M A V L S L P N 20

226 Y I A I S R Y I H K S S R Q F 20

37 H N E F D T I V L P V L Y L I 19

44 V L P V L Y L I I F V A s I L 19

73 K T S F I F Y L K N I V V A D 19

217 A V L V I L I G c Y I A I S R 19

45 L P V L Y L I I F V A S I L L 18

65 W I F F H I R N K T S F I F Y 18

113 L C R Y T s V L F Y A N M Y T 18

154 S I T F T K V L S V C V V I 18

171 V L S L P N I I L T N G Q P T 18

267 P Y H L C R I P F T F S H L D 18

277 F S H L D R L L D E S A Q K I 18

75 S F I F Y L K N I V V A D L I 17

89 I M T L T F P F R I V H D A G^' 17

106 P W Y F K F I L C R Y T S V L 17

119 V L F Y A N M Y T S I V F L G 17

206 A V T Y V N S C L F V A V L V 17

212 S C L F V A V L V I L I G C Y 17

46 P V L Y L I I F V A S I L L N 16

50 L I I F V A S I L L N G L A V 16

61 G L A V W I F F H I R N K T s 16

63 A V W I F F H I R N K T S F I 16

91 T L T F P F R I V H D A G F G 16

93 T F P F R I V H D A G F G P w 16

128 S I V F L G L I S I D R Y L K 16

163 V C V W V I M A V L S L P N I 16

238 R Q F I S Q S S R K R K H N Q 16

134 L I S I D R Y L K V V K P F G 15

143 V V K P F G D S R M Y S I T F 15 TABLE XXVI - 154P2A8 HLA- DRB1*1101 15 - mers

Pos ^■ 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Seq. ID Num.

151 R M Y S I T F T K V L S V C V 15

164 C V W V I M A V L S L P N I I 15

196 K S P L G V K W H T A V T Y V 15

204 H T A V T Y V N S C L F V A V 15

251 N Q S I R V V V A V F F T C F 15

288 A Q K I L Y Y C K E I T L F L 15

TABLE XXIII - 156P1D4 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

140 V P I I I I F G V I F C I I 35

3 W L L F F L V T A I H A E L c 33

47 E Y L F K A M V A F s M R K V 33

122 T L E F L K I P S T L A P P M 33

80 R V S F F V V T D P S K N H 31

96 L P A V E V Q S A I R M N K N- 30

187 E N M I T I E N G I P S D P L 28

28 R L S I R T A L G D K A Y A 27

55 A F S M R K V P N R E A T E I 27

148 G V I F C I I I V A I A L L I 27

155 I V A I A L L I L S G I W Q R 27

157 A I A L L I L s G I W Q R,R R 27

58 M R K V P N R E A T E I S H V 26

123 L E F L K I P s T L A P P M D 26

13 H A E L C Q P G A E N A F K V 25

139 S V P I w I I I F G V I F C I 25

149 V I F c I I I V A I A L L I L 25

150 I F C I I I V A I A L L I L S 25

90 P S K N H T L P A V E V Q S A 24

114 N A F F L N D Q T L E F L K I 24

125 F L K I P S T L A P P M D P S 24

144 I I I F G V I F C I I I V A I 24

151 F C I I I V A I A L L I L S G 24

2 L L L F f L V T A I H A E L 23

6 F F L V T A I H A E L C Q P G 23

71 H V L L C N V T Q R V S F W F 23

74 L C N V T Q R V S F F V V T 23

93 N H T L P A V E V Q S A I R M 23

115 A F F L N D Q T. L E F L K I P 23

152 C I I I V A I A L L I L S G I 23

186 C E N M I T I E N G I P S D P 23

66 A T E I S H V L L C N V T Q R 22

70 S H V L L C N V T Q R V S F W 22

133 A P P M D P S V P I W I I I F 22

143 I I I F G V I F c I I I V A 22

147 F G V I F C I I I V A I A L L 22

189 M I T I E N G I P s D P L D M 22

197 P S D P L D M K G G H I N D A 22

26 K V R L S I R T A L G D K A Y 21

107 M N K N R I N N A F F L N D Q 21

190 I T I E N G I P S D P L D M K 21

24 A F K V R L s I R T A L G D K 20

130 S T L A P P M D P s V P I W I 20

39 A Y A D T N E Ξ Y L F K A M 19

45 N E E Y L F K A M V A F S M R 19 TABLE XXIII - 156P1D4 HLA- DRB1*0101 15 - mers

Seq. ID

Pos • 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

50 F K A M V A F S M R K V P N R 19

104 A I R M N K N R I N N A F F L 19

153 I I I V A I A L L I L S G I W 19

1 M L w L L F F L V T A I H A E 18

10 T A I H A E L C Q P G A E N A 18

23 N A F K V R L s I R T A L G D 18

46 E E Y L F K A M V A F S M R K 18

51 K A M V A F S M R K V P N R E 18

82 S F W F V V T D P S K N H T L 18

117 F L N D Q T L E F L K I P S T 18

198 S D P L D M K G G H I N D A F 18

5 L F F L V T A I H A E L C Q P 17

16 L C Q P G A E N A F K V R L S 17

22 E N A F K V R L S I R T A L G 17

44 T N E E Y L F K A M V A F S M 17

112 I N N A F F L N D Q T L E F L 17

120 D Q T L E F L K I P s T L A P 17

142 I W I I I F G V I F c I I I V 17

146 I F G V I F C I I I V A I A L 17

200 P L D M K G G H I N D A F M T 17

TABLE XXIV - 156P5C12 HLA- DRB1*0301 (DR17 ) 15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

139 L F H L S V D S E H R R Q G I 27

42 T L I L L L G G P L A L L L V 24

91 C T D M S D I T K S Y L S E R 24

190 s M G F K K T G Q S F F C V W 24

43 L I L L L G σ P L A L L L V S 23

50 P L A L L L V S G S W L L A L 22

51 L A L L L V s G S W L L A L V 22

129 D P T L R E K R L Q L F H L S 22

170 Y S E V I L D T G T I Q L S A 22

34 R Q L L K L P R T L I L L L G 21

36 L L K L P R T L I L L L G G P 21

40 P R T L I L L L G G P L A L L 21

44 I L L L G G P L A L L L V S G 21

65 V F S I S L F P A L W F L A K 21

122 V G A L P V D D P T L R E K R 21

171 s E V I L D T G T I Q L S A M 21

177 T G T I Q L s A M A L Y Q S M 21

22 s R G M A E H A P A T F R Q L 20

33 F R Q L L K L P R T L I L L L 20

53 L L L V S G S W L L A L V F S 20

87 D M T L C T D M S D I T K S Y 20

204 W A R L V A L H T V H F I Y H 20

59 s W L L A L V F s I S L F P A 19

74 L W F L A K K P w T E Y V D M 19

90 L C T D M S D I T- ^K S Y L S E 19

155 K A L V R T V L Q F A R D Q G 19

159 R T V L Q F A R D Q G Y S E V 19

182 L S A M A L Y Q S M G F K K T 19

15 Q V V G L L S R G M A E H A 18

63 A L V F S I S L F P A L W F L 18

108 C F W V A E S E E K V V G M V 18

128 D D P T L R E K R L Q L F H L 18 TABLE XXIV - 156P5C12 HLA- DRB1*0301 (DR17) 15 - mers

Pos • 1 2 Seq. ID

3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

158 V R T V L Q F A R D Q G Y S E 18

161 V L Q F A R D Q G Y S E V I L 18

3 P C H ϊ R K Y Q E S D R Q W V 17

7 R K Y Q E S D R Q W V V G L L 17

30 P A T F R Q L L K L P R T L I 17

141 H L S V D S E H R R Q G I A K 17

26 A E H A P A T F R Q L L K L P 16

71 F P A L W F L A K K P W T E Y 16

164 F A R D Q G Y S Ξ V I L D T G 16

17 V V G L L S R G M A E H A P A 15

41 R T L I L L L G G P L A L L L 15

52 A L L L V S G S W L L A L V F 15

151 Q G I A K A L V R T V L Q F A 15

198 Q S F F C V W A R L V A L H T 15

126 P V D D P T L R E K R L Q L F 14

67 S I S L F P A L W F L A K K p^' 13

115 E E K V V G M V G A L P V D D 13

116 E K V V G M V G A L P V D D P 13

118 V V G M V G A L P V D D P T L^' 13

123 G A L P V D D P T L R E K R L 13

200 F F C V W A R L V A L H T V H 13

TABLE XXV - 156P5C12 HLA- DRB1*0401 (DR4Dw4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

106 G S C F w V A E S E E K V V G 28

14 R Q V V G L L s R G M A E H 26

15 Q V V G L L S R G M A E H A 26

33 F R Q L L K L P R T L I L L L 26

59 S L L A L V F S I S L F P A 26

87 D M T L C T D M S D I T K s Y 26

210 L H T V H F I Y H L P S S K V 26

213 V H F I Y H L P S S K V G s L 26

6 I R K Y Q E S D R Q w V V G L 22

57 S G S L L A L V F s I S L F 22

68 I s L F P A L W F L A K K P 22

72 P A L W F L A K K P T E Y V 22

107 S C F V A E S E E K V V G M 22

197 G Q S F F C V W A R L V A L H 22

198 Q S F F C V A R L V A L H T 22

201 F C V A R L V A L H T V H F 22

34 R Q L L K L P R T L I L L L G 20

40 P R T L I L L L G G P L A L L 20

41 R T L I L L L G G P L A L L L 20

48 G G P L A L L L V S G S L L 20

50 P L A L L L V S G S W L L A L 20

51 L A L L L V S G S w L L A L V 20

58 G S L L A L V F s I S L F P 20

62 L A L V F S I S L F P A L W F 20

67 S I S L F P A L W F L A K K P 20

91 C T D M S D I T K s Y L S E R 20

116 E K V V G M V G A L P V D D P 20

141 H L s V D S E H R R Q G I A K 20

150 R Q G I A K A L V R T V L Q F 20

155 K A L V R T V L Q F A R D Q G 20

158 V R T V L Q F A R D Q G Y S E 20

170 Y S E V I L D T G T I Q L S A 20 TABLE XXV - 156P5C12 HLA- DRB1*0401 (DR4DW4) 15 - mers

Seq. ID

Pos . 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

171 S E V I L D T G T I Q L S A M 20

204 W A R L V A L H T V H F I Y H 20

7 R K Y Q E S D R Q w V V G L L 18

20 L L S R G M A E H A P A T F R 18

27 E H A P A T F R Q L L K L P R 18

81 P W T E Y V D M T L C T D M S 18

84 E Y V D M T L C T D M s D I T 18

128 D D P T L R E K R L Q L F H L 18

133 R E K R L Q L F H L S V D S E 18

151 Q G I A K A L V R T V L Q F A 18

152 G I A K A L V R T V L Q F A R 18

169 G Y S E V I L D T G T I Q L S 18

30 P A T F R Q L L K L P R T L I 16

63 A L V F S I S L F P A L w F L 16

79 K K P W T E Y V D M T L C T D 16

98 T K S Y L S E R G S C F w V A 16

161 V L Q F A R D Q G Y S E V I L 16

167 D Q G Y S E V I L D T G T I Q 16

185 M A L Y Q S M G F K K T G Q S 16

190 S M G F K K T G Q s F F C V 16

212 T V H F I Y H L P s s K V G s 16

188 Y Q S M G F K K T G Q s F F c 15

18 V G L L S R G M A Ξ H A P A T 14

36 L L K L P R T L I L L i. G G P 14

42 T L I L L L G G P L A L L L V 14

43 L I L L L G G P L A L L L V s 14

44 I L L L G G P L A L L L V S G 14

52 A L L L V S G S W L L A L V F 14

53 L L L V S G S W L L A L V F S 14

61 L L A L V F S I S L F P A L w 14

65 V F S I S L F P A L W F L A K 14

71 F P A L F L A K K P W T E Y 14

83 T E Y V D M T L C T D M s D I 14

108 C F W V A E S E E K V V G M V 14

118 V V G M V G A L P y D D P T L 14

119 V G M V G A L P V D D P T L R 14

122 V G A L P V D D P T L R E K R 14

124 A L P V D D P T L R E K R L Q 14

134 Ξ K R L Q L F H L S V D S E H 14

136 R L Q L F H L S V D S E H R R 14

139 L F H L S V D S E H R R Q G I 14

154 A K A L V R T V L Q F A R D Q 14

159 R T V L Q F A R D Q G Y S E V 14

172 E V I L D T G T I Q L •s A M A 14

179 T I Q L S A M A L Y Q s M G F 14

182 L S A M A L Y Q S M G F K K T 14

184 A M A L Y Q S M G F K K T G Q 14

205 A R L V A L H T V H F I Y H L 14

207 L V A L H T V H F I Y H L P S 14

TABLE XXIV - 156P1D4 HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

114 N A F F L N D Q T L E F L K I ^' 29 24 A F K V R L S I R T A L G D K 27. 37 D K A Y A W D T N E E Y L F K 24 TABLE XXIV - 156P1D4 HLA- DRB1*0301 (DR17 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

83 F W F V V T D P S K N H T L P 23 193 E N G I P S D P L D M K G G H 22 150 I F C I I I V A I A L L I L S 21

5 L F F L V T A I H A E L c Q P 20

152 C I I I V A I A L L I L S G I 20

74 L C N V T Q R V S F w F V V T 19

84 W F V V T D P S K N H T L P A 19

102 Q S A I R M N K N R I N N A F 19

55 A F S M R K V P N R E A T E I 18

98 A V E V Q S A I R M N K N R I 18

115 A F F L N D Q T L E F L K I P 18

175 E P S E V D D A E D K C E N M 18

186 C E N M I T I E N G I P S D P 18

14 A E L C Q P G A E N A F K V R 17

105 I R M N K N R I N N A F F L N 17

163 L S G I w Q R R R K N K E P S 17

196 I P S D P L D M K G G H I N D 17

39 A Y A W D T N E E Y L F K A M 16

51 K A M V A F S M R K V P N R E 16

71 H V L L C N V T Q R V S F W F 16

85 F V V T D P S K N H T L P A V 16

113 N N A F F L N D Q T L E F L K 16

160 L L I L S G I W Q R R R K N K 16

18 Q P G A E N A F K V R L S I R 15

72 V L L C N V T Q R V S F w F V 15

106 R M N K N R I N N A F F L N D 15

201 L D M K G G H I N D A F M T E 15

1 M L W L L F F L V T A I H A E 14

31 I R T A L G D K A Y A w D T N 14 38 K A Y A W D T N E Ξ Y L F K A 14 123 L E F L K I P S T L A P P M D 14 159 A L I L S G I W Q R R R K N 14

TABLE XXV - 156P1D4 HLA- DRB1*0401 (DR4DW4 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

80 R V S F W F V V T D P S K N H 28

122 T L E F L K I P S T L A P P M 28

5 L F F L V T A I H A E L C Q P 26

55 A F S M R K V P N R E A T E I 26

71 H V L L C N V T Q R V S F W F 26

98 A V E V Q S A I R M N K N R I 26

3 W L L F F L V T A I H A E L C 22

37 D K A Y A W D T N E Ξ Y L F K 22

81 V S F W F V V T D P S K N H T 22

114 N A F F L N D Q T L E F L K I 22

144 I I I F G V I F c I I I V A I 22

148 G V I F C I I I V A I A L L I 22

1 M L W L L F F L V T A I H A E 20

2 L W L L F F L V T A I H A E L 20

24 A F K V R L S I R T A L G D K 20 28 R L S I R T A L G D K A Y A W 20 46 E E Y L F K A M V A F S M R K 20 50 F K A M V A F S M R K V P N R 20 66 A T E I S H V L L C N V T Q R 20 70 S H V L L C N V T Q R V S F W 20 83 F W F V V T D P S K N H T L P 20 00 a tO tO H H H H H H H H H H £ vo cD iπ H _{ω m} £ cD rf to vo cn cn cn ιn ιf* H H H H H H H H H H H H H H H H H a o o O O -J rf* rf* rf* l0 t0 t0 _0 H O o oo cD cn ιπ tfi tfι ιπ ιπ iπ rf* ιf* _o to o 0 cn iπ o ω cn tO H Lo vo tfi io in vo ° c rf* H W ^{ω m} <2 £ M -j g rf* co ι to vo vo ιn o v ιo ~α en o -o. tfi _o _ H θ -j _o vo o to to

a H Ω a TJ H H TJ uj a a a < a s a a < a a < a cn a B H rt a a a a a B rt B B Ω a H H Ω a H Ά si cn α KD a H a

< to rt Ώ a cn a S H TJ cn a a a H J a rt a cn a a K a ri tr H rt H a a a a a a a a a H < H H Ω a Ω t-t ιO co a to a ω cn ω a a a Ω H Si a rt a Ά a a a > s a > a Ώ H i a H a a n a a o < a Ω H Ω 3 a H > H H H Ω H TJ H > rt H lo * a rf_* a a H 3 H H 3 a H a a H 3 < a H <J H s; a Ά a a B a a B a KD a H H 3 a a H H H H H H H tr H a O a a <_ < rf_* fr_*

Si in H a a H H H O T

* X a a a a a rt Ω rt ; H a < a a a a > > < rt > a 3 TJ a rt H 01 t > < H H TJ a s; B a a O in H a σi H o Ω a a •fl H TJ a cn H o a a < a a KD KD H a < Ω 3 a a a rt a <! Ώ a cn H rt Ω H IT H <J H Ω Ω H >τj 2 H cn < H 1 O Ω Ω Ω a co TJ rt a o a iO a cn a B H a H a rt s a H o ⁱτ] is a H a B > a α a H H a a H < H < H tr a < O H -o a

< 1 t a a cn cn 3 Ω a a α o < a j*

00 a x B H < Ω < 3 a co r TJ TI a < Ω O H a B Ω mi a a s; co H a H H H H « a a co

H o rt vo in H tfl 3 H O H H < a α rt a a H a Ω a Q O ) cn co ω a KD a a co rt cn a ιo a Ω a D Ω a p F fr H H •A a H a < vo rf_*

O o 1 en a rt a a H •fl H H T a a B a a H a a B o a a < rt cn •fl H o < ϋ ω < < X a s a α H Ώ a H co H - f < Ω Ω a a ιo O a H

H 3 B o Ω < Ω a s: cn a p a a a a <: a a H a a co a TJ TJ KD > rt a < H a a Ω 3 a H a s; Ω a H tr a H < cn H cn H fl) O H in cn f_* t Hi r o a H Ω H 3 a a ϋ > 3 rt TJ Ω Ω a a a a 3 rt a H rt H a a co a a a cn a a iO H cn a H a H H H 1-3 a to O 1 a a

LO a B a a H H H H J α a a ιo a a a a a O H Ω a a a o ≤ t* rϋ Pi Ω a x < a Ω o n a a s; Ω co a H pa H a a a H ω 3 a it* > 1 a 3 3 H H H H TJ 3 H H a a a a a a H rt a a 3 H B 3 < a Ω < a a a Ω a o a a KD H Ω cn a a < Ω a ID rf* a tfl a ι-3 H a < H a s; cn o a a a a a a Ω ω >-. n t* < Ω a H •A a n < a a a a > a a a a a a i H Ω cn tr H TJ •fl 3 in co to H H H H H H H H H H H H H H H H H H H H H H H H H H HH HH HH HH HH HH HH HH HH HH HH ttOO ttOO t_OO ttO tt tO tO tO t tO t t _0 _O tO t o rf_* o rf* rf* ιf» ιf* rf* rf* rf* rf rf* !t rf* !f» !f* rf* rf* !t* rf* ιf* ιi ιn en cn cn cn ι rom crDri CmD CD em» co cmo røm m m m "i - --- r-- _-» r O-- O--- <O-^ .O-* O^ rO-- -O-- -O-- 0 0 0 O

Hi fl)

cn a ro

3 ?^■?

" H o

TABLE XXVI - 156P1D4 HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

3 W L L F F L V T A I H A E L C 23

122 T L E F L K I P S T L A P P M 23

148 G V I F C I I I V A I A L L I 23

51 K A M V A F s M R K V P N R E 21

71 H V L L C N V T Q R V S F W F 20

55 A F S M R K V P N R E A T E I 19

164 S G I W Q R R R K N K E P s E 19

186 C E N M I T I E N G I P S D P 19

47 E Y L F K A M V A F s M R K V 18

81 V S F W F V V T D P s K N H T 18

93 N H T L P A V E V Q s A I R M 18

144 I I I F G V I F C I I I V A I 18

63 N R E A T E I S H V L L c N V 17

140 V P I W I I I F G V I F c I I 17

2 L W L L F F L V T A I H A E L 16

84 W F V V T D P S K N H T L P A 16

101 V Q S A I R M N K N R I N N A 16

6 F F L V T A I H A E L c Q P G 15

52 A M V A F S M R K V P N R E A 15

86 V V T D P S K N H T L P A V E 15

160 L L I L S G I W Q R R R K N K 15

5 L F F L V T A I H A E L C Q P 14

24 A F K V R L S I R T A L G D K 14 30 S I R T A L G D K A Y A W D T 14 43 D T N E E Y L F K A M V A F s 14 98 A V E V Q S A I R M N K N R I 14 119 N D Q T L E F L K I P S T L A 14 143 W I I I F G V I F c I I I V A 14 163 L S G I W Q R R R K N K E P S 14 166 I W Q R R R K N K E P s E V D 14 187 E N M I T I E N G I P s D P L 14 22 E N A F K V R L S I R T A L G 13 23 N A F K V R L S I R T A L G D 13 25 F K V R L S I R T A L G D K A 13 28 R L S I R T A L G D K A Y A W 13 46 E E Y L F K A M V A F S M R K 13 130 S T A P P M D P S V P I W I 13 139 S V P I W I I I F G V I F C I 13 146 I F G V I F C I I I V A I A L 13 147 F G V I F c I I I V A I A L L 13 150 I F C I I I V A I A L L I L S 13 152 C I I I V A I A L L I L s G I 13 153 I I I V A I A L L I L S G I w 13 9 V T A I H A E L C Q P G A E N 12

10 T A I H A E L C Q P G A E N A 12 13 H A Ξ L C Q P G A E N A F K V 12 66 T E I S H V L L C N V T Q R 12 82 S F W F V V T D P S K N H T L 12 104 A I R M N K N R I N N A F F L 12 113 N N A F F L N D Q T L E F L K 12 120 D Q T L E F L K I P S T L A P 12 123 L E F L K I P S T L A P P M D 12 137 D P S V P I W I I I F G V I F 12 154 I I V A I A L L I L S G I w Q 12 155 I V A I A L L I L S G I w Q R 12 157 A I A L L I L S G I W Q R R R 12 184 D K C E N M I T I E N G I P S 12 189 M I T I E N G I P S D P L D M 12 TABLE XXVI - 156P1D4 HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

195 G I P S D P L D M K G G H I N 12

200 P L D M K G G H I N D A F M T 12

4 L L F F L V T A I H A E L C Q 11

39 A Y A W D T N E E Y L F K A M 11

161 L I L S G I W Q R R R K N K E 11

TABLE XXIII - 156P5C12 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

14 R Q W V V G L L S R G M A E H 34

41 R T L I L L L G G P L A L L L 34

40 P R T L I L L L G G P L A L L 33

197 G Q S F F C V W A R L V A L H 33

116 E K V V G M V G A L P V D D P 32

50 P L A L L L V S G S w L L A L ^' 31

115 E E K V V G M V G A L P V D D 31

204 W A R L V A L H T V H F I Y H 31

213 V H F I Y H L P S S K V G s L 31

59 S W L L A L V F S I S L F P A 29

33 F R Q L L K L P R T L I L L L 27

48 G G P L A L L L V s G S W L L 27

98 T K S Y L S E R G s C F W V A 26

169 G Y S E V I L D T G T I Q L S 26

201 F C V W A R L V A L H T V H F 26

15 Q W V V G L L S R G M A E H A 25

65 V F S I S L F P A L W F L A K 25

182 L S A M A L Y Q S M G F K K T 25

18 V G L L S R G M A E H A P A T 24

34 R Q L L K L P R T L I L L L G 24

42 T L I L L L G G P L A L L L V 24

44 I L L L G G P L A L L L V S G 24

55 L V S G S W L L A L V F S I S 24

58 G S W L L A L V F S I S L F P 24

61 L L A L V F S I S L F P A L W 24

119 V G M V G A L P V D D P T L R 24

175 L D T G T I Q L S A M A L Y Q 24

176 D T G T I Q L S A M A L Y Q S 24

51 L A L L L V S G S W L L A L V 23

171 S E V I L D T G T I Q L S A M 23

188 Y Q S M G F K K T G Q S F F C 23

210 L H T V H F I Y H L P S S K V 23

11 E S D R Q W V V G L L S R G M 22

22 S R G M A E H A P A T F R Q L 22

62 L A L V F S I S L F P A L w F 22

84 E Y V D M T L C T D M S D I T 22

91 c^'τ D M S D I T K S Y L S E R 22

179 T I Q L S A M A L Y Q S M G F 22

190 S M G F K K T G Q S F F C V W 22

39 L P R T L I L L L G G P L A L 21

63 A L V F s I S L F P A L W F L 21

30 P A T F R Q L L K L P R T L I 20

107 S C F W V A E S E E K V V G M 20

57 S G S W L L A L V F S I S L F 19

79 K K P W T E Y V D M T L C T D 19

96 D I T K S Y L S E R G S C F W 19

146 S E H R R Q G I A K A L V R T 19

150 R Q G I A K A L V R T V L Q F 19 TABLE XXIII - 156P5C12 HLA- DRB1*0101 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

185 M A L Y Q S M G F K K T G Q S 19 6 I K Y Q E S D R Q W V V G L 18

68 I S L F P A L W F L A K K P W 18 70 L F P A L W F L A K K P W T E 18 71 F P A L W F L A K K P W T E Y 18 83 T E Y V D M T L C T D M S D I 18 108 C F W V A E S E E K V V G M V 18 113 E S E E K V V G M V G A L P V 18 129 D P T L R E K R L Q L F H L s 18 145 D S E H R R Q G I A K A L V R 18 151 Q G I A K A L V R T V L Q F A 18 161 V L Q F A R D Q G Y S E V I L 18 170 Y S E V I L D T G T I Q L S A 18 189 Q S M G F K K T G Q s F F C V 18 198 Q S F F C V W A R L V A L H T 18 52 A L L L V S G S W L L A L V F 17 56 V S G S W L L A L V F S I S L 17 75 W F L A K K P W T E Y V D M T 17 82 W T E Y V D M T L C T D M S D 17 92 T D M S D I T K S Y L S E R G 17 106 G S C F W V A E S E E K V V G 17 126 P V D D P T L R E K R L Q L F 17 131 L R E K R L Q L F H L S V D 17 132 L R E K R L Q L F H L S V D S 17 154 A K A L V R T V L Q F A R D Q 17 157 L V R T V L Q F A R D Q G Y S 17 162 L Q F A R D Q G Y S E V I L D 17 184 A M A L Y Q S M G F K K T G Q 17 202 C V W A R L V A L H T V H F I 17 205 A R L V A L H T V H F I Y H L 17 9 Y Q E S D R Q W V V G L L S R 16

19 G L L S R G M A Ξ H A P A T F 16

32 T F R Q L L K L P R T L I L L 16

36 L L K L P R T L I L L L G G P 16

43 L I L L L G G P L A L L L V S 16

45 L L L G G P L A L L L V S G S 16

53 L L L V S G S W L L A L V F S 16

67 S I S L F P A L W F L A K K P 16

73 A L W F L A K K P W T E Y V D 16

112 A E S E E K V V G M V G A L P 16

133 R E K R L Q L F H L S V D S Ξ 16

136 R L Q L F H L S V D S E H R R 16

155 K A L V R T V L Q F A R D Q G 16

167 D Q G Y S E V I L D T G T I Q 16

172 E V I L D T G T I Q L S A M A 16

212 T V H F I Y H L P S S K V G S 16

TABLE XXVI - 156P5C12 HLA- DRB1*1101 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Seq. ID Num.

15 Q W V V G L L S R G M A E H A 26 198 Q S F F C V W A R L V A L H T 26 210 L H T V H F I Y H L P S S K V 26

30 P A T F R Q L L K L P R T L I 24 167 D Q G Y S E V I L D T G T I Q 22

33 F R Q L L K L P R T L I L L L 21

58 G S W L L A L V F S I S L F P 21 TABLE XXVI - 156P5C12 HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

115 E E K V V G M V G A L P V D D 21

141 H L S V D S E H R R Q G I A K 21

71 F P A L W F L A K K P W T E Y 20

91 C T D M S D I T K S Y L s E R 20

158 V R T V L Q F A R D Q G Y S E 20

13 D R Q W V V G L L S R G M A E 19

41 R T L I L L L G G P L A L L L 19

62 L A L V F S I S L F P A L W F 19

72 P A W F L A K K P W T E Y V 19

197 G Q S F F C V W A R L V A L H 19

6 I R K Y Q E S D R Q W V V G L 18

48 G G P L A L L L V S G S W L L 18

119 V G M V G A L P V D D P T L R 18

179 T I Q L S A M A L Y Q S M G F 18

201 F C V W A R L V A L H T V H F 18

151 Q G I A K A L V R T V L Q F A 17

185 M A L Y Q S M G F K K T G Q S 17

68 I S L F P A L W F L A K K P W 16

82 W T E Y V D M T L C T D M s D 16

11 E S D R Q W V V G L L S R G M 15

109 F W V A E S E E K V V G M V G 15

112 A E S E E K V V G M V G A L P 15

147 E H R R Q G I A K A L V R T V 15

20 L L S R G M A E H A P A T F R 14

38 K L P R T L I L L L G G P L A 14

67 S I S L F P A L W F h A K K P 14

79 K K P W T E Y V D M T L C T D 14

133 R E K R L Q L F H L S V D s E 14

154 A K A L V R T V L Q F A R D Q 14

186 A L Y Q S M G F K K T G Q S F 14

14 R Q W V V G L L S R G M A Ξ H 13

37 L K L P R T L I L L L G G P L 13

52 L L L V S G s W L L A L V F 13

96 D I T K S Y L s E R G S C F W 13

113 E S E E K V V G M V G A L P V 13

136 R L Q L F H L S V D S E H R R 13

172 E V I L D T G T I Q L S A M A 13

213 V H F I Y H L P S S K V G S L 13

18 V G L L S R G M A E H A P A T 12

19 G L L S R G M A E H A P A T F 12

39 L P R T L I L L L G G P L A L 12

40 P R T L I L L L G G P L A L L 12

47 L G G P L A L L L V S G S W L 12

50 P L L L V S G S W L L A L 12

56 V S G S W L L A L V F S I S L 12

59 S W L L A L V F S I S L F P A 12

61 L L A L V F S I S L F P A L W 12

84 E Y V D M T L c T D M S D I T 12

116 E K V V G M V G A L P V D D P 12

127 V D D P T L R E K R L Q L F H 12

137 L Q L F H L S V D S E H R R Q 12

155 K A L V R T V L Q F A R D Q G 12

181 Q L S A M A L Y Q S M G F K K 12

188 Y Q S M G F K K T G Q S F F c 12

204 W A R L V A L H T V H F I Y H 12

207 L V A L H T V H F I Y H L P S 12 TABLE XXIII - 159P2B5 HLA- DRB1*0101 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

145 P L F W G I L G G V F F L G S 38

94 S C T L V A L L S L G L Q D P 33

144 W P L F W G I L G G V F F L G 32

68 S R S V L A M R A I L P P S L 30

71 V L A M R A I L P P S L S K S 30

10 R P T F W G W A A T P A P V S 27

13 F W G W A A T P A P V S A P G 26

40 P P G G G F L G S T S F Q R R 25

72 A M R A I L P P S L S K S V 25

97 L V A L L S L G L Q D P L G C 25 ^•

151 L G G V F F L G S R A C T R T 25

175 A A L L R L L F P A P R R P G 25

11 P T F W G W A A T P A P V S A 24

56 E K E L L W E R G Q D V S R S 24

65 Q D V S R S V L A M R A I L P 24

91 L P H s C T L V A L L S L G L 24

102 S L G L Q D P L G C R A P A T 24

119 T P A G A T L S A S S L P R P 24

148 W G I L G G V F F L G S R A c 24

174 A A L L R L L F P A P R R P 24

177 L L R L L F P A P R R P G A R 24

75 R A I L P P S L S K S V H F P 23

100 L L S L σ L Q D P L G c R A P 23

135 S P S A s L L L S W P L F W G 23

88 F P P L P H S C T L V A L L S 22

127 A S S L P R P C S P S A S L L 22

48 S T S F Q R R G E K E L L W Ξ 20

179 R L L F P A P R R P G A R S R 20

112 R A P A T K P T P A G A T L S 19

115 A T K P T P A G A T L S A S S 19

141 L L S W P L F W G I L G G V F 19

34 G E R Q G S P P G G G F L G S 18

79 P P S L S K S V H F P P L P H 18

85 S V H F P P L P H S C T L V A 18

103 L G L Q D P L G C R A P A T K 18

107 D P L G C R A P A T K P T P A 18

139 S L L L S W P L F W G I L G G 18

143 S W P L F W G I L G G V F F Ii 18

150 I L G G V F F L G S R A c T R 18

153 G V F F L G S R A C T R T Q A 18

164 R T Q A R R H T G P A A A L L 18

187 R P G A R S R A G Y A S P G S 18

TABLE XXIV - 159P2B5 HLA- DRB1*0301 (DR17 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

98 V A L L S L G L Q D P L G C R 26 48 S T S F Q R R G Ξ K E L L w E 23

94 S C T L V A L L S L G L Q D P 21 71 V L A M R A I L P P S L S K S 20 57 K E L L W E R G Q D V S R S V 18 178 L R L L F P A P R R P G A R S 18 20 P A P V S- A P G N P P T G E G 17 49 T S F Q R R G E K E L L W E R 17 67 V S R S V L A M R A I L P P S 17 144 W P L F W G I L G G V F F L G 17 152 G G V F F L G S R A c T R T Q 17 TABLE XXIV - 159P2B5 HLA- DRB1* 0301 - (DR17 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

171 T G P A A A L L R L L F P A P 17

185 P R R P G A R S R A G Y A S P 16

2 V K R Ξ H G Q E R P T F W G W 15

61 W E R G Q D V S R S V L A M R 15

74 M R A I L P P S L S K S V H F 15

97 L V A L L s L G L Q D P L G C 15

137 S A S L L L S W P L F W G I L 14

138 A S L L L s W P L F W G I L G 14

174 A A A L L R L L F P A P R R P 14

179 R L L F P A P R R P G A R S R 14

204 R R S P G T A H K G S L P W P 14

43 G G F L G S T S F Q R R G E K 13

56 E K E L L w E R G Q D V S R S 13

68 S R S V L A M R A I L P P S L 13

95 C T L V A L L S L G L Q D P L 13

101 L S L G L Q D P L G C R A P A 13

136 P S A s L L L s W P L F W G I 13

147 F W G I L G G V F F L G S R A 13

175 A A L L R L L F P A P R R P G 13

69 R S V L A M R A I L P P S L S 12

75 R A I L P P S L s K S V H F P 12

100 L L s L G L Q D P L G C R A P 12

151 L G G V F F L G s R A C T R T 12

177 L L R L L F P A P R R P G A R 12

TABLE XXV - 159P2B5 HLA- DRB1*0401 (DR4DW4 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

85 s V H F P P L P H S C T L V A 28

94 s C T L V A L L S L G L Q D P 26

11 P T F W G W A A T P A P V S A 22

152 G G V F F L G S R A C T R T Q 22

179 R L L F P A P R R P G A R S R 22

^'20 P A P V S A P G N P P T G E G 20

56 Ξ K E L. L W E R G Q D V S R S 20

71 V L A M R A I L P P S L S K S 20

75 R A I L P P s L S K S V H F P 20

88 F P P L P H s C T L V A L L S 20

95 C T L V A L L s L G L Q D P L 20

98 V A L L S L G L Q D P L G C R 20

127 A S S L P R P C S P S A S L L 20

148 W G I L G G V F F L G S R A C 20

151 L G G V F F L G S R A C T R T 20

178 L R L L F P A P R R P G A R S 20

60 L W E R G Q D V S R S V L A M 18

61 W E R G Q D V s R S V L A M R 18

67 V S R S V L A M R A I L P P S 18

124 T L S A S S L P R P C S P S A 18

135 S P S A s L L L S W P L F W G 18

155 F F L G s R A C T R T Q A R R 18

10 R P T F W G W A A T P A P V S 16

13 F W G W A A T P A P V S A P G 16

42 G G G F L G s T S F Q R R G E 16

141 L L S W P L F W G I L G G V F 16

144 W P L F W G I L G G V F F L G 16

145 P L F W G I L G G V F F L G S 16

193 R A G Y A s P G S P E R R S P 16 TABLE XXV - 159P2B5 HLA- DRB1*0401 (DR4DW4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

57 K E L L W E R G Q D V S R S V 15 74 M R A I L P P S L S K S V H F 14 79 P P S L S K S V H F P P L P H 14 83 S K S V H F P P L P H S C T L 14 97 L V A L L S L G L Q D P L G C 14 102 S L G L Q D P L G C R A P A T 14 139 S L L L S W P L F W G I L G G 14 143 S W P L F W G I L G G V F F L 14 174 A A L L R L L F P A P R R P 14 175 A A L L R L L F P A P R R P G 14

TABLE XXVI - 159P2B5 HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

85 S V H F P P L P H S C T L V A 24

179 R L L F P A P R R P G A R S R 24

79 P P S L S K S V H F P P L P H 22

178 L R L L F P A P R R P G A R S 22

124 T L S A S S L P R P C S P S A 20

68 S R S V L A M R A I L P P S L 19

152 G G V F F L G S R A C T R T Q 19

97 L V A L L S L G L Q D P L G C 18

148 W G I L G G V F F L G S R A C 18

174 A A A L L R L L F P A P R R P 18

13 F W G W A A T P A P V S A P G 17

141 L L S W P L F w G I L G G V F 17

144 W P L F W G I L G G V F F L G 17

10 R P T F W G w A A T P A P V S 16

11 P T F W G W A A T P A P V S A 16

145 P L F W G I L G G V F F L G S 16

161 A C T R T Q A R R H T G P A A 16

183 P A P R R P G A R S R A G Y A 16

61 W E R G Q D V S R S V L A M R 15

76 A I L P P s L S K S V H F P P 15

91 L P H S C T L V A L L S L G L 15

147 F W G I L G G V F F L G S R A 15

171 T G P A A A L L R L L F P A P 15

177 L L R L L F P A P R R P G A R 15

196 Y A S P G S P E R R S P G T A 15

28 N P P T G E G E R Q G S P P G 14

53 R R G Ξ K E L L W E R G Q D V 14

65 Q D V S R S V L A M R A I L P 14

102 S L G L Q D P L G C R A P A T 14

104 G L Q D P L G C R A P A T K P 14

109 L G C R A P A T K P T P A G A 14

122 G A T L S A S S L P R P C S P 14

151 L G G V F F L G S R A C T R T 14

156 F L G S R A C T R T Q A R R H 14

162 C T R T Q A R R H T G P A A A 14

197 A S P G S P E R Ri S P G T A H 14

203 E R R S P G T A H K G S L P W 14

57 K E L L W E R G Q D V S R S V 13

72 L M R A I L P P S L S K S V 13

74 M R A I L P P S L S K S V H F 13

88 F P P L P H S C T L V A L L S 13

95 C T L V A L L S L G L Q D P L 13

99 A L L S L G L Q D P L G C R A 13 TABLE XXVI - 159P2B5 HLA- DRB1*1101 15 - mers

Seq. ID

Pos • 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

103 L G L Q D P L G C R A P A T K 13

209 T A H K G S L P W P L A L R L 13

69 R S V L A M R A I L P P S L S 12

71 V L A M R A I L P P S L S K S 12

83 S K S V H F P P L P H S c T L 12

94 S C T L V A L L s L G L Q D P 12

127 A S S L P R P C s P S A S L L 12

134 C S P S A S L L L S W P L F W 12

138 A S L L L S W P L F W G I L G 12

172 G P A A A L L R L L F P A P R 12

175 A A L L R L L F P A P R R P G 12

45 F L G S T S F Q R R G E K E L 11

TABLE XXIII - 161P2B7a HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

90 V A P Y V N V G A L R M P F Q 34

142 G L P L A T L A A D S A S A A 32

138 A P P F G L P L A T L A A D S 28

74 H K G V L I G A A s Q F E A C 27

130 H A P Y M M F P A P P F G L P 27

37 D A F M R E E L S Q R L G L S 24

72 Q L H K G V L I G A A S Q F E 24

100 R M P F Q Q V Q A Q L Q L D S 24

120 H H H L H P H L A A H A P Y M 24

121 H H L H P H L A A H A P Y M M 24

132 P Y M M F P A P P F G L P L A 24

145 L A T L A A D S A S A A S V V 24

103 F Q Q V Q A Q L Q L D S A V A 22

106 V Q A Q Q L D S A V A H A H 22

117 A H A H H H L H P H L A A H A 22

124 H P H L A A H A P Y M M F P A 22

139 P P F G L P L A T L A A D S A 22

152 S A S A A S V V A A A A A A K 22

153 A S A A S V V A A A A A A K T 22

129 A H A P Y M M F P A P P F G L 21

15 R T N F T L E Q L N E L E R L 20

70 E N Q L H K G V L I G A A S Q 20

174 I A D L R L K A K K H A A A L 20

45 s Q R L G L S E A R V Q V W F 19

56 Q V W F Q N R R A K C R K Q E 19

3 D E G Q T K I K Q R R S R T N 18

20 L E Q L N E L E R L F D E T H 18

27 E R L F D E T H Y P D A F M R 18

87 A C R V A P Y V N V G A L R M 18

91 A P Y V N V G A L R M P F Q Q 18

133 Y M M F -P A P P F G L P L A T 18

173 S I A D L R L K A K K H A A A 18

42 E E L S Q R L G L S E A R V Q 17

66 C R K Q E N Q L H K G V L I G 17

67 R K Q E N Q L H K G V L. I G A 17

68 K Q E N Q L H K G V L I G A A 17

73 L H K G V L I G A A S Q F E A 17

79 I G A A S Q F Ξ A C R V A P Y 17

107 Q A Q L Q L D S A V A H A H H 17

154 S A A S V V A A A A A A K T T 17 TABLE XXIII - 161P2B7a HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

155 A A S V V A A A A A A K T T S 17 156 A S V V A A A A A A K T T S K 17 161 A A A A A K T T S K N S S I A 17 171 N S S I A D L R L K A K K H A 17 176 D L R L K A K K H A A A L G L 17 6 Q T K I K Q R R S R T N F T L 16

17 N F T L Ξ Q L N E L E R L F D 16 23 L N E L E R L F D E T H Y P D 16 25 E L E R L F D E T H Y P D A F 16 41 R E E L S Q R L G L S E A R V 16 44 L S Q R L G L S E A R V Q V W 16 54 R V Q V W F Q N R R A K C R K 16 76 G V L I G A A S Q F E A C R V 16 83 S Q F Ξ A C R V A P Y V N V G 16 93 Y V N V G A L R M P F Q Q V Q 16 104 Q Q V Q A Q L Q L D S A V A H 16 109 Q L Q L D S A V A H A H H H L 16 123 L H P H L A A H A P Y M M F P 16 134 M M F P A P P F G L P L A T L 16 137 P A P P F G L P L A T L A A D 16 141 F G L P L A T L A A D S A S A 16 147 T L A A D S A S A A S V V A A 16 148 L A A D S A S A A S V V A A A 16

TABLE XXIV - 161P2B7a HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

107 Q A Q L Q L D S A V A H A H H 24

37 D A F M R E E L S Q R L G L S 22 145 L A T L A A D S A s A A S V V 22

74 H K G V L I G A A s Q F E A C 20 132 P Y M M F P A P P F G L P L A 20 170 K N S S I A D L R L K A K K H 20

15 R T N F T L E Q L N E L E R L 19

20 L E Q L N E L E R L F D E T H 19

96 V G A L R M P F Q Q V Q A Q L 19

19 T L E Q L N E L E R L F D E T 18

87 A C R V A P Y V N V G A L R M 18 103 F Q Q V Q A Q L Q L D S A V A 18 174 I A D L R L K A K K H A A A L 18

17 N F T L E Q L N E L E R L F D 17

52 E A R V Q V W F Q N R R A K C 17

54 R V Q V W F Q N R R A K C R K 17

76 G V L I G A A S Q F E A C R V 17

82 A S Q F E A C R V A P Y V N V 17 7 T K I K Q R R S R T N F T L E 16 9 I K Q R R S R T N F T L E Q L 16

24 N E L Ξ R L F D E T H Y P D A 16

39 F M R Ξ E L S Q R L G L S E A 16

92 P Y V N V G A L R M P F Q Q V 16

48 L G L S E A R V Q V W F Q N R 15

62 • R R A K C R K Q E N Q L H K G 15

99 L R M P F Q Q V Q A Q L Q L D 15 101 M P F Q Q V Q A Q L Q L D S A 15

33 T H Y P D A F M R E E L S Q R 14

67 R K Q E N Q L H K G V L I G A 14 176 D L R L K A K K H A A A L G L 14 TABLE XXIV - 161P2B7a HLA- DRB1*0301 (DR17 ) 15 - mers os Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

25 E L E R L F D E T H Y P D A F 13 20 H H H L H P H L A A H A P Y M 13 40 P F G L P L A T L A A D S A S 13 55 A A S V V A A A A A A K T T S 13 41 R E E L S Q R L G L S E A R V 12 45 S Q R L G L S E A R V Q V W F 12 47 R L G L S E A R V Q V W F Q N 12 75 K G V L I G A A S Q F E A C R 12 98 A L R M P F Q Q V Q A Q L Q L 12 09 Q h Q L D S A V A H A H H H L 12 56 A S V V A A A A A A K T T S K 12 6 Q T K I K Q R R S R T N F T L 11

16 T N F T L E Q L N E L E R L F 11 31 D E T H Y P D A F M R E E L S 11 13 D S A V A H A H H H L H P H L 11 38 A P P F G L P L A T L A A D S 11 66 K T T S K N S S I A D L R L K 11 71 N S S I A D L R L K A K K H A 11

TABLE XXV - 161P2B7a HLA- DRB1*0401 (DR4DW4) 15 - mers

Seq. ID os 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

37 D A F M R E Ξ L S Q R L G L S 26

76 G V L I G A A S Q F E A C R V 26

15 R T N F T L E Q L N E L E R L 22

90 V A P Y V N V G A L R M P F Q 22 00 R M P F Q Q V Q A Q L Q L D S 22

20 L E Q L N E L E R L F D E T H 20

26 L E R L F D E T H Y P D A F M 20

54 R V Q V W F Q N R R A K C R K 20

75 K G V L I G A A S Q F E A C R 20

87 A C R V A P Y V N V G A L R M 20

93 Y V N V G A L R M P E Q Q V Q 20

96 V G A L R M P F Q Q V Q A Q L 20 03 F Q Q V Q A Q L Q L D S A V A 20 13 D S A V A H A H H H L H P H L 20 32 P Y M M F'P A P P F G L P L A 20 42 G L P L A T L A A D S A S A A 20 45 L A T L A A D S A S A A S V V 20 55 A A S V V A A A A A A K T T S 20 56 A S V V A A A A A A K T T S K 20 71 N S S I A D L R L K A K K H A 20

3 D E G Q T K I K Q R R S R T N 18

33 T H Y P D A F M R E E L S Q R 18 46. Q R L G L S E A R V Q V W F Q 18 53 A R V Q V W F Q N R R A K C R 18 64 A K C R K Q E N Q L H K G V L 18 97 G A L R M P F Q Q V Q A Q L Q 18 06 V Q A Q L Q L D S A V A H A H 18 39 P P F G L P L A T L A A D S A 18 44 P L A T L A A D s A S A A S V 18 49 A A D S A S A A s V V A A A A 18 59 V A A A A- A A K T T s K N S S 18 70 K N S S I A D L R L K A K K H 18 27 E R L F D E T Ή Y P D A F M R 16 32 E T H Ύ P D A F M R E E L S Q 16 36 P D A F M R E E L S Q R L G L 16 TABLE XXV - 161P2B7a HLA- DRB1*0401 (DR4D 4 ) 15 - mers

Pos q. ID

1 2 3 4 5 6 7 8 9 0 1 2 Se

3 4 5 score Num.

55 V Q V W F Q N R R A K C R K Q 16

130 H A P Y M M F P A P P F G L P 16

138 A P P F G L P L A T L A A D s 16

6 Q T K I K Q R R S R T N F T L 15

176 D L R L K A K K H A A A L G L 15

17 N F T L E Q L N E L E R L F D 14

23 L N E L E R L F D E T H Y P D 14

45 S Q R L G L S E A R V Q V W F 14

47 R L G L S E A R V Q V W F Q N 14

52 E A R V Q V W F Q N R R A K C 14

70 E N Q L H K G V L I G A A s Q 14

98 A L R M P F Q Q V Q A Q L Q L 14

107 Q A Q L Q L D S A V A H A H H 14

109 Q L Q L D S A V A H A H H H L 14

120 H H H L H P H L A A H A P Y M 14

124 H P H L A A H A P Y M M F P A 14

140 P F G L P L A T L A A D S A S 14

5 G Q T K I K Q R R S R T N F T 12

8 K I K Q R R S R T N F T L E Q 12

11 Q R R S R T N F T L E Q L N E 12

12 R R S R T N F T L E Q L N E L 12

14 S R T N F T L E Q L N E L E R 12

25 E L E R L F D E T H Y P D A F 12

38 A F M R E E L S Q R L G L S Ξ 12

39 F M R Ξ E L S Q R L G L S E A 12

42 E E L S Q R L G L S E A R V Q 12

44 L S Q R L G L S E A R V Q V W 12

49 G L S E A R V Q V W F Q N R R 12

58 W F Q N R R A K C R K Q E N Q 12

66 C R K Q E N Q L H K G V L I G 12

67 R K Q E N Q L H K G V L I G A 12

71 N Q L H K G V L I G A A s Q F 12

72 Q L H K G V L I G A A S Q F E 12

73 L H K G V L I G A A S Q F E A 12

79 I G A A S Q F Ξ A C R V A P Y 12

80 G A A S Q F E A C R V A P Y V 12

81 A A S Q F E A C R V A P Y V N 12

84 Q F Ξ A C R V A P Y V N V G A 12

85 F Ξ A C R V A P Y V N V G A L 12

88 C R V A P Y V N V G A L R M P 12

92 P Y V N V G A L R M P F Q Q V 12

99 L R M P F Q Q V Q A Q L Q L D 12

101 M P F Q Q V Q A Q L Q L D S A 12

104 Q Q V Q A Q L Q L D S A V A H 12

110 L Q L D S A V A H A H H H L H 12

112 L D S A V A H A H H H L H P H 12

116 V A H A H H H L H P H L A A H 12

117 A H A H H H L H P H L A A H A 12

118 H A H H H L H P H L A A H A P 12

122 H L H P H L A A H A P Y M M F 12

123 L H P H L A A H A P Y M M F P 12

135 M F P A P P F G L P L A T L A 12

137 P A P P F G L P L A T L A A D 12

143 L P L A T L A A D S A S A A S 12

146 A T L A A D S A S A A S V V A 12

147 T L A A D S A S A A S V V A A 12

150 A D S A S A A S V V A A A A A 12

152 S A S A A S V V A A A A A A K 12 TABLE XXV - 161P2B7a HLA- DRB1*0401 (DR4DW4 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

153 A S A A S V V A A A A A A K T 12 154 S A A S V V A A A A A A K T T 12 157 S V V A A A A A A K T T S K N 12 158 V V A A A A A A K T T S K N S 12 161 AA A A K T T S K N s S I A 12 163 A A A K T T S K N s S I A D L 12 168 T S K N S S I A D L R L K A K 12 169 S K N S s I A D L R L K A K K 12 175 A D L R L K A K K H A A A L G 12

TABLE XXVI - 161P2B7a HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

20 L Ξ Q L N E L E R L F D Ξ T H 20 55 V Q V W F Q N R R A K C R K Q 20 174 I A D L R L K A K K H A A A L 20 176 D L R L K A K K H A A A L G L 20 90 V A P Y V N V G A L R M P F Q 18 142 G L P L A T L A A D S A S A A 18

3 D E G Q T K I K Q R R S R T N 17

100 R M P F Q Q V Q A Q L Q L D S 17

133 Y M M F P A P P F G L P L A T 17

138 A P P F G L P L A T L A A D s 17

27 E R L F D Ξ T H Y P D A F M R 16

26 L E R L F D E T H Y P D A F M 15

54 R V Q V W F Q N R R A K C R K 15

67 R K Q E N Q L H K G V L I G A 15

81 A A S Q F E A C R V A P Y V N 15

113 D S A V A H A H H H L H P H L 15

170 K N S s I A D L R L K A K K H 15

38 A F M R E E L S Q R L G L S E 14

70 E N Q L H K G V L I G A A S Q 14

87 A C R V A P Y V N V G A L R M 14

91 A P Y V N V G A L R M P F Q Q 14

110 L Q L D S A V A H A H H H L H 14

114 S A V A H A H H H L H P H L A 14

118 H A H H H L H P H L A A H A P 14

152 S A S A A S V V A A A A A A K 14

158 V V A A A A A A K T T S K N s 14

162 A AA A K T T S K N S S I A D 14

175 A D L R L K A K K H A A A L G 14

34 H Y P D A F M R E E L S Q R L 13

72 Q L H K G V L I G A A S Q F E 13

106 V Q A Q L Q L D S A V A H A H 13

117 A H A H H H L H P H L A A H A 13

153 S A A S V V A A A A A A K T 13

15 R T N F T L E Q L N E L E R L 12

17 N F T L E Q L N Ξ L E R L F D 12

23 L N E L E R L F D E T H Y P D 12

42 E E L s Q R L G L S E A R V Q 12

71 N Q L H K G V L I G A A S Q F 12

76 G V L I G A A S Q F E A C R V 12

93 Y V N V G A L R M P F Q Q V Q 12

103 F Q Q V Q A Q L Q L D S A V A 12

104 Q Q V Q A Q L Q L D S A V A H 12

120 H H H L H P H L A A H A P Y M 12

121 H H L H P H L A A H A P Y M M 12 TABLE XXVI - 161P2B7a HLA- DRB1*1101 15 - mers

Seq. ID

Pos ■ 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

128 A A H A P Y M M F P A P P F G 12 129 A H A P Y M M F P A P P F G L 12 131 A P Y M M F P A P P F G L P L 12 139 P P F G L P L A T L A A D S A 12 145 L A T L A A D S A S A A S V V 12 171 N S S I A D L R L K A K K H A 12 32 E T H Y P D A F M R E E L S Q 11 2 E D E G Q T K I K Q R R S R T 10 5 G Q T K I K Q R R S R T N F T 10

36 P D A F M R E E L S Q R L G L 10 56 Q W F Q N R R A K C R K Q E 10 57 V w F Q N R R A K C R K Q E N 10 82 A S Q F E A C R V A P Y V N V 10 130 H A P Y M M F P A P P F G L P 10 4 E G Q T K I K Q R R s R T N F 9

52 E A R V Q V W F Q N R R A K C 9 53 A R V Q V W F Q N R R A K C R 9 164 A A K T T S K N S S I A D L R 9 172 S S I A D L R L K A K K H A A 9

TABLE XXIII - 179P3G7 HLA- DRB1*0101 15 - mers

Seq. ID os 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

28 S A G M Y M Q S G S D F N C G 31 72 R C P Y IJ. K H Q T L E L E K E 27

22 G E R Y S R S A G M Y M Q S G 26 46 P S Y Y R A S P S Y S A D K 26 86 H L E S P Q L G G K V S F P E 26

85 A Y R L E Q P V G R P L S s C 25 93 G G K V S F P E T P K S D s Q 25

38 D F N C G V M R G C G L A P S 24

40 N C G V M R G C G L A P S L S 24

70 P S Y L S Q L D S W G D P K A 24 45 V P S Y Y R A S P S Y S A L D 24 71 E A P F E Q R A s L N P R A E 24 17 E Q s L A G P K G S P S E s Ξ 24

63 S L A L N T Y P S Y L S Q L D 23 81 N P R A E H L E s P Q L G G K 23 85 E H L E S P Q L G G K V S F P 23

43 V M R G C G L A P S L S K R D 22

60 S S P S L A L N T Y P S Y L S 22

89 E Q P V G R P L s S c-s Y P P 22 22 S G P E A A L Y s H P L P E S 22

83 K A A Y R L E Q P V G R P L S 21 08 T P s P N E I K T E Q S L A G 21 07 E E N V C C M Y s A E N R A K 20

41 C G V M ^■R G C G L A P S L S K 19 11 C C M Y S A E N R A K S G P E 19 52 S P S Y S A L D K T P H C s G 19 15 K T E Q S L A G P K G S P s E 19 27 P S E S Ξ K E R A K A A D s S 19 60 G N W L T A K S G R K K R c P 19

9 P N S Y A E P A A P G G G E 18

10 N S Y A E P L A A P G G G E R 18 20 G G G E R Y S R S A G M Y M Q 18 50 A P S L S K R D E G S S P S L 18 TABLE XXI II - 179P3G7 HLA- DRB1 * 0101 15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

51 P S L S K R D E G s S P S L A 18

73 L S Q L D S W G D P K A A Y R 18

76 L D S W G D P K A A Y R L E Q 18

115 S A E N R A K S G P E A A L Y 18

167 A N D F E A P F E Q R A S L N 18

195 K V S F P E T P K S D S Q T P 18

230 S E K E R A K A A D S S P D T 18

250 K E E I K A Ξ N T T G N w L T 18

257 N T T G N W L T A K S G R K K 18

280 T L E L E K E F L F N M Y L T 18

4 P R N V T P N S Y A E P L A A 17

13 A E P L A A P G G G E R Y S R 17

29 A G M Y M Q S G S D F N C G V 17

52 S L S K R D E G S S P S L A L 17

102 P P S V K E Ξ N V C C M Y S A 17

116 A E N R A K S G P E A A L Y S 17

132 P L P E S C L G E H E V P V P 17

135 E S C L G E H E V P V P S Y Y 17

159 D K T P H C S G A N D F E A P 17

212 N E I K T E Q S L A G P K G s 17

214 I K T E Q S L A G P K G S P s 17

277 K H Q T L E L E K E F L F N M 17

278 H Q T L E L E K E F L F N M Y 17

282 E L E K E F L F N M Y L T R E 17

5 R N V T P N S Y A E P L A A P 16

42 G V M R G C G L A P S L S K R 16

66 L N T Y P S Y L S Q L D S W G 16

67 N T Y P s Y L S Q L D S W G D 16

69 Y P S Y L S Q L D S W G D P K 16

77 D S W G D P K A A Y R L E Q P 16

90 Q P V G R P L S S C S Y P P S 16

93 G R P L S s C S Y P P S V K E 16

119 R A K S G P E A A L Y S H P L 16

125 E A A L Y s H P L P E S C L G 16

126 A A L Y S H P L P E S c L G E 16

127 A L Y S H P L P E S C L G E H 16

140 E H E V P V P S Y Y R A S P S 16

142 E V P V P S Y Y R A S P S Y S 16

143 V P V P S Y Y R A S P S Y S A 16

176 Q R A S L N P R A E H L E S P 16

198 F P E T P K S D S Q T P S P N 16

219 S L A G P K G S P S E S E K E 16

229 E S E K E R A K A A D S S P D 16

252 E I K A E N T T G N W L T A K 16

259 T G N W L T A K S G R K K R C 16

7 V T P N S Y A E P L A A P G G 15

18 A P G G G E R Y s R S A G M Y 15

46 G c G L A P S L s K R D E G S 15

74 S Q L D S W G D P K A A Y R L 15

86 Y R L E Q P V G R P L S S C S 15

174 F E Q R A S L N P R A E H L E 15

191 Q L G G K V S F P Ξ T P K S D 15

211 P N E I K T E Q s L A G P K G 15

256 E N T T G N W L T A K S G R K 15

TABLE XXIV - 179P3G7 HLA-

DRB1 *0301 (DR17 ) 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Seq . ID Num .

75 Q L D S W G D P K A A Y R L E 27

278 H Q T L E L E K Ξ F L F N M Y 27

163 H C S G A N D F E A P F E Q R 25

28 S A G M Y M Q S G S D F N C G 20

240 S S P D T S D N E A K E E I K 20

61 s P S L A L N T Y P S Y L S Q 19

110 V C C M Y S A E N R A K S G P 19

117 E N R A K S G P E A A L Y S H 19

199 P E T P K s D S Q T P S P N E 19

85 A Y R L E Q P V G R P L S S C 18

142 E V P V P s Y Y R A s P S Y S 18

177 R A S L N P R A E H L E S P Q 18

277 K H Q T L E L E K E F L F N M 18

32 Y M Q S G S D F N C G V M R G 17

187 L E S P Q L G G K V s F P E T 17

276 T K H Q T L Ξ L E K E F L F N 17

50 A P S L S K R D E G S S P S L 16

127 A L Y S H P L P E S C L G E H 16

195 K V s F P E T P K S D S Q T P 16

167 A N D F E A P F E Q R A S L N 15

169 D F E A P F E Q R A S L N P R 15

209 P S P N E I K T Ξ Q S L A G P 15

264 T A K S G R K K R C P Y T K H 15

4 P R N V T P N s Y A E P L A A 14

282 E L E K E F L F N M Y L T R E 14

30 G M Y M Q S G S D F N C G V M 13

41 C G V M R G C G L A P s L S K 13

73 L S Q L D S W G D P K A A Y R 13

211 P N E I K T E Q S L A G P K G 13

260 G N W L T A K S G R K K R c P 13

TABLE XXV - 179P3G7 HLA- DRB1*0401 (DR4D 4 ) 15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

111 c C M Y S A E N R A K S G P E 28

259 T G N W L T A K S G R K K R C 28

110 V C C M Y S A E N R A K S G P 26

250 K E E I K A Ξ N T T G N W L T 26

29 A G M Y M Q S G S D F N c G V 22

145 V P S Y Y R A S P S Y S A L D 22

146 P S Y Y R A S P s Y S A L D K 22

272 R C P Y T K H Q T L Ξ L E K Ξ 22

63 S L A L N T Y P s Y L S Q L D 20

70 P S Y L S Q L D s W G D P K A 20

89 Ξ Q P V G R P L s S C S Y P P 20

102 P P S V K E E N V C C M Y s A 20

107 E E N V C C M Y s A E N R A K 20

193 G G K V S F P E T P K S D S Q 20

211 P N E I K T E Q s L A G P K G 20

280 T L E L Ξ K E F L F N M Y L T 20

32 Y M Q S G S D F N C G V M R G 18

44 M R G C G L A P S L S K R D E 18

52 S L S K R D E G S S P S L A L 18

60 S S P S L A L N T Y P s Y L S 18

67 N T Y P S Y L S Q L D s W G D 18

79 W G D P K A A Y R L E Q P V G 18

86 Y R L E Q P V G R P L S S C S 18

122 S G P E A A L Y S H P L P E S 18

168 N D F E A P F E Q R A S L N P 18

169 D F E A P F E Q R A S L N P R 18 TABLE XXV - 179P3G7 HLA- DRB1*0401 (DR4D 4 ) 15 - mers

Seq. ID

POS 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

178 A S L N P R A E H L E S P Q L 18

181 N P R A E H L E S P Q L σ G K 18

199 P E P K S D S Q T P S P N E 18

208 T P S P N E I K T E Q S L A G 18

233 E R A K A A D S s P D T s D N 18

282 E L E K E F L F N M Y L T R E 18

22 G E R Y S R S A G M Y M Q S G 16

66 L N T Y P S Y L s Q L D s w G 16

83 K A A Y R L E Q P V G R P L S 16

98 S C S Y P P S V K E E N V C C 16

152 S P S Y S A L D K T P H C S G 16

167 A N D F E A P F E Q R A s L N 16

195 K V S F P E T P K S D S Q T P 16

41 C G V M R G C G L A P S L S K 14

46 G C G L A P s L S K R D E G S 14

73 L S Q L D S w G D P K A A Y R 14

85 A Y R L E Q P V G R P L S S C 14

93 G R P L S S c S Y P P S V K E 14

125 E A A L Y S H P L P E S c L G 14

140 E H E V P V P S Y Y R A S P S 14

142 E V P V P S Y Y R A S P s Y S 14

155 Y S A L D K T P H C S G A N D 14

184 A E H L E S P Q L G G K V S F 14

217 E Q S L A G P K G S P S Ξ S E 14

278 H Q T L E L E K E F L F N M Y 14

TABLE XXVI - 179P3G7 HLA- DRB1*1101 15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

111 C C M Y S A E N R A K S G P E 25 152 S P S Y S A L D K T P H C S G 24 195 K V S F P E T P K s D S Q T P 24 86 Y R L E Q P V G R P L S S c S 21 37 S D F N C G V M R G C G L A P 20 98 S C S Y P P S V K E E N V C C 20 142 E V P V P s Y Y R A S P S Y S 20 155 Y S A L D K T P H C S G A N D 20 167 A N D F E A P F E Q R A S L N 18 9 P N S Y A E P L A A P G G G E 16

69 Y P S Y L s Q L D S W G D P K 16 145 V P S Y Y R A S P S Y S A L D 16 146 P S Y Y R A S P S Y S A L D K 16 171 E A P F E Q R A S L N P R A E 16 224 K G s P S E S E K E R A K A A 16 47 C G L A P S L S K R D Ξ G S S 15 123 G P E A A L Y S H P L P E S C 15 133 L P E S C L G E H E V P V P S 15 140 E H E V P V P S Y Y R A S P S 15 226 S P S E S E K E R A K A A D S 15 263 L T A K S G R K K R C P Y T K 15 4 P R N V T P N S Y A E P L A A 14

19 P G G G E R Y s R S A G M Y M 14 48 G L A P S L S K R D E G S S P 14 113 M Y S A E N R A K S G P E A A 14 178 A S L N P R A E H L E s P Q L 14 216 T E Q S L A G P K G S P S E S 14 277 K H Q T L E L E K E F L F N M 14 TABLE XXVI - 179P3G7 HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

36 G S D F N C G V M R G C G L A 13

43 V M R G C G L A P S L S K R D 13

63 S L A L N T Y P S Y L s Q L D 13

73 L S Q L D S W G D P K A A Y R 13

82 P K A A Y R L E Q P V G R P L 13

135 E S C L G E H E V P V P S Y Y 13

186 H L E S P Q L G G K V S F P E 13

193 G G K V S F P E T P K S D S Q 13

214 I K T E Q S L A G P K G S P S 13

10 N S Y A E P L A A P G G G E R 12

25 Y S R S A G M Y M Q S G S D F 12

40 N C G V M R G C G L A P S L S 12

41 C G V M R G C G L A P s L S K 12

50 A P S L S K R D E G S s P S L 12

60 S S P S L A L N T Y P s Y L S 12

67 N T Y P S Y L S Q L D s W G D 12

70 P s Y L S Q L D S W G D P K A 12

89 E Q P V G R P L S S C S Y P P 12

93 G R P L S S C S Y P P s V K E 12

104 S V K E E N V C C M Y s A E N 12

107 E E N V C C M Y S A E N R A K 12

125 E A A L Y S H P L P E S C L G 12

208 T P S P N E I K T E Q S L A G 12

211 P N E I K T E Q s L A G P K G 12

217 E Q S L A G P K G S P S E S E 12

257 N T T G N W L T A K S G R K K 12

259 T G N W L T A K S G R K K R C 12

272 R C P Y T K H Q T L E L E K E 12

TABLE XXIII - 184P3C10B HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

140 Q L R L L F L V G T A S N P H 34

11 A T L I L A I G A F T L L L F 31

190 V L F L Q W Q E T R C A N A S 30

239 I Q N V G P I R A F w S K Y Y 30

236 G Q L I Q N V G P I R A F W S 29

17 I G A F T L L L F S L L V S P 28

33 T C K V Q E Q P P A I P E A L 28

105 Q P V F L L L V I K s S P S N 28

107 V F L L L V I K s S P S N Y V 28

89 C R H F P L L Q D V P P S K C 27

133 E R K V R G L Q L R L L F L V 27

179 H D S F F N L T L K Q V L F L 27

264 Y P P Y C G G G G F L L S R F 27

8 R P N A T L I L A I G A F T L 26 104 A Q P V F L L L V I K S S P S 26 130 W G R E R K V R G L Q L R L L 26 142 R L L F L V G T A S N P H E A 26 22 L L L F S L L V s P P T C K V 25 46 A L A W P T P P T R P A P A P 25 156 R K V N R L L E L E A Q T H 25 159 V N R L L E L E A Q T H G D I 25 231 R H L F V G Q L I Q N V G P I 25 21 T L L L F S L L V S P P T C K 24 25 F S L L V S P P T C K V Q E Q 24 218 D N M V F Y L Q D H D P G R H 24 TABLE XXIII - 184P3C10B HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

254 V P E V V T Q N E R Y P P Y C 24

272 G F L L S R F T A A A L R R A 24

18 G A F T L L L F S L L V S P P 23

106 P V F L L L V I K S S P S N Y 23

110 L L V I K S S P S N Y V R R E 23

141 L R L L F L V G T A S N P H E 22

162 L L E L E A Q T H G D I L Q W 22

271 G G F L L S R F T A A A L R R 22

92 F P L L Q,D V P P S K C A Q P 21

138 G L Q L R L L F L V G T A S N 21

202 N A S F V L N G D D D V F A H 21

12 T L I A I G A F T L L L F S 20

83 F L L Y R H C R H F P L L Q D 20

148 G T A S P H E A R K V N R L 20

180 D S F F N L T L K Q V L F L Q 20

246 R A F W S K Y Y V P E V V T Q 20

251 K Y Y V P E V V T Q N E R Y P 20

261 N E R Y P P Y C G G G G F L L 20

270 G G G F L L S R F T A A A L R 20

275 L S R F T A A A L R R A A H V 20

3 Y L R H R R P N A T L I L A I 19

9 P N A T L I L A I G A F T L L 19

123 R E L L R R T W G R E R K V R 19

262 E R Y P P Y C G G G G F L L S 19

108 F L L L V I K S S P S N Y V R 18

182 F F N L T L K Q V L F L Q W Q 18

192 F L Q W Q E T R C A N A S F V 18

195 W Q E T R C A N A S F V L N G 18

211 D D V F A H T D N M V F Y L Q 18

227 H D P G R H L F V G Q I Q N 18

255 P E V V T Q N E R Y P P Y C G 18

10 A T L I L A I G A F T L L L 17

19 A F L L L F S L L V S P P T 17

20 F T L L F S L L V S P P T C 17

66 S M V H P D F A.T Q P Q H V 17

81 Q N F L L Y R H C R H F P L L 17

136 V R G L Q L R L L F L V G T A 17

173 I L Q W D F H D S F F N L T L 17

187 L K Q V L F L Q W Q E T R C A 17

204 S F V L N G D D D V F A H T D 17

219 N M V F Y L Q D H D P G R H L 17

222 F Y L Q D H D P G R H L F V G 17

228 D P G R H L F V G Q L I Q N V 17

233 L F V G Q L I Q N V G P I R A 17

234 F V G Q L I Q N V G P I R A F 17

242 V G P I R A F W S K Y Y V P E 17

263 R Y P P Y C G G G G F L L S R 17

269 G G G G F L L S R F T A A A L 17

278 F T A A A L R R A A H V L D I 17

1 M K Y L R H R R P N A T L I L 16

4 L R H R R P N A T L I L A I G 16

14 I L A I G A F T L L L F S L L 16 16 A I G A F T L L L F S L L V S 16 30 S P P T C K V Q E Q p p^' A I P 16 31 P P T C K V Q E Q P P A I P E 16 40 P P A I P E A L A W P T P P T ^' 16 41 P A I P E A L A W P T P P T R 16 42 A I P E A L A W P T P P T R P 16 TABLE XXIII - 184P3C10B HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

49 W P T P P T R P A P A P C H A 16

55 R P A P A P C H A N T s M V T 16

70 H P D F A T Q P Q H V Q N F L 16

82 F L L Y R H C R H F P L L Q 16

88 H C R H F P L L Q D V P P S K 16

96 Q D V P P S K C A Q P V F L L 16

109 L L L V I K S s P S N Y V R R 16

135 K V R G L Q L R L L F L V G T 16

163 L E L E A Q T H G D I L Q W D 16

170 H G D I L Q W D F H D S F F N 16

171 G D I L Q W D F H D S F F N L 16

181 S F F N L T L K Q V L F L Q W 16

193 L Q W Q E T R C A N A S F V L 16

201 A A S F V L N G D D D V F A 16

203 A S F V L N G D D D V F A H T 16

209 G D D D V F A H T D N M V F Y 16

210 D D D V F A H T D N M V F Y L 16

232 H L F V G Q L I Q N V G P I R 16

235 V G Q L I Q N V G P I R A F W 16

268 C G G G G F L L s R F T A A A 16

279 A L R R A A H V L D I F 16

284 R R A A H V L D I F P I D D V 16

TABLE XXIV - 184P3C10B HLA- DRB1*0301 (DR17 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

171 G D I L Q W D F H D S F F N L 28 180 D S F F N L T L K Q V L F L Q 25 91 H F P L L Q D V P P S K C A Q 22 211 D D V F A H T D N M V F Y L Q 22 11 A T L I L A I G A F T L L L F 21 109 L L L V I K s S P S N Y V R R 21 182 F F N L T L K Q V L F L Q W Q 21 204 S F V L N G D D D V F A H T D 21 14 I L A I G A F T L L L F S L L 20 17 I G A F T L L L F S L L V S P 20 81 Q N F L L Y R H C R H F P L L 20 133 E R K V R G L Q L R L L F L V 20 174 L Q W D F H D S F F N L T L K 20 203 A S F V L N G D D D V F A H T 20 272 G F L L S R F T A A A L R R A 20 12 T L I L A I G A F T L L L F S 19 106 P V F L L L V I K S S P S N Y 19 181 S F F N L T L K Q V L F L Q W 19 212 D V F A H T D N M V F Y L Q D 19 222 F Y L Q D H D P G R H L F V G 19 242 V G P I R A F W S K Y Y V P E 19 134 R K V R G L Q L R L L F L V G 18 156 R K V N R L L E L E A Q T H 18 217 T D N M V F Y L Q D H D P G R 18 255 P E V V T Q N E R Y P P Y C G 18 132 R E R K V R G L Q L R L L F L 17 144 L F L V G T A S N P H E A R K 17 160 N R L L E L E A Q T H G D I L 17 188 K Q V L F L Q W Q E T R C A N 17 232 H L F V G Q L I Q N V G P I R 17 TABLE XXIV - 184P3C10B HLA- DRB1* 0301 (DR17 ) 15 - mers

Seq. :

Pos ^• 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

254 V P E V V T Q N E R Y P P Y C 17

2 K Y L R H R R P N A T L I L A 16

82 N F L L Y R H c R H F P L L Q 16

176 W D F H D S F F N L T L K Q V 16

189 Q V L F L Q W Q E T R C A N A 16

194 Q W Q Ξ T R C A N A S F V L N 16

223 Y L Q D H D P G R H L F V G Q 16

19 A F T L L L F S L L V S P P T 15

70 H P D F A T Q P Q H V Q N F L 15

73 F A T Q P Q H V Q N F L L Y R 15

74 A T Q P Q H V Q N F L L Y R H 15

115 S S P S N Y V R R E L L R R T 15

152 N P H E A R K V N R L L Ξ L Ξ 15

159 V N R L L E L E A Q T H G D I 15

167 A Q T H G D I L Q W D F H D s 15

253 Y V P E V V T Q N E R Y P P Y 15

271 G G F L L S R F T A A A L R R 15

20 F T L L L F S L L V S P P T C 14

25 F s L L V S P P T C K V Q E Q 14

97 D V P P S K c A Q P V F L L L 14

110 L L V I K S s P S N Y V R R E 14

114 K S S P S N Y V R R E L L R R 14

1 M K Y L R H R R P N A T L I L 13

10 N A T L I L A I G A F T L L L 13

24 L F S L L V S P P T C K V Q E 13

33 T C K V Q E Q P P A I P E A L 13

64 N T S M V T H P D F A T Q P Q 13

95 L Q D V P P S K C A Q P V F L 13

104 A Q P V F L L L V I K S s P S 13

107 V F L L L V I K S S P s N Y V 13

136 V R G L Q L R L L F L V G T A 13

138 G L Q L R L L F L V G T A s N 13

140 Q L R L L F L V G T A s N P H 13

141 L R L L F L V G T A S N P H E 13

184 N L T L K Q V L F L Q W Q E T 13

187 L K Q V L F L Q W Q E T R C A 13

251 K Y Y V P E V V T Q N E R Y P 13

280 A A A L R R A A H V L D I F P 13

TABLE XXV - 184P3C10B HLA- DRB1*0401 (DR4Dw4 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

22 L L L F S L L V S P P T C K V 28

107 V F L L L V I K S S P S N Y V 26

141 L R L L F L V G T A S N P H E 26

17 I G A F T L L L F S L L V S P 22

46 A L A W P T P P T R P A P A P 22

70 H P D F A T Q P Q H V Q N F L 22

83 F L L Y R H C R H F P L L Q D 22

105 Q P V F L L L V I K S S P S N 22

173 I L Q W D F H D s F F N L T L 22

179 H D S F F N L T L K Q V L F L 22

250 S K Y Y V P E V V T Q N E R Y 22

275 L S R F T A A A L R R A A H V 22

10 N A T L I L A I G A F T L L L 20

11 A T L ϊ L A I G A F T L L L F 20

19 A F T L L L F S L L V S P P T 20 TABLE XXV - 184P3C10B HLA- DRB1*0401 (DR4DW4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

25 F S L L V S P P T C K V Q E Q 20

40 P P A I P E A L A W P T P P T 20

91 H F P L L Q D V P P S K c A Q 20

104 A Q P V F L L L V I K s s P S 20

110 L L V I K S S P S N Y V R R E 20

118 S N Y V R R E L L R R T W G R 20

133 E R K V R G L Q L R L L F L V 20

138 G L Q L R L L F L V G T A S N 20

140 Q L R L L F L V G T A s N P H 20

144 L F L V G T A S N P H E A R K 20

156 A R K V N R L L E L E A Q T H 20

160 N R L L E L E A Q T H G D I L 20

162 L L E L E A Q T H G D I L Q W 20

171 G D I L Q W D F H D S F F N L 20

187 L K Q V L F L Q W Q Ξ T R C A 20

190 V L F L Q W Q E T R C A N A S 20

204 S F V L N G D D D V F A H T D 20

232 H L F V G Q L I Q N V G P I R 20

236 G Q L I Q N V G P I R A F w S 20

239 I Q N V G P I R A F W S K Y Y 20

242 V G P I R A F W S K Y Y V P E 20

251 K Y Y V P Ξ V jV T Q N E R Y P 20

272 G F L L S R F T A A A L R R A 20

280 A A A L R R A A H V L D I F P 20

13 L I L A I G A F T L L L F S L 18

18 G A F T L L L F S L L V S P P 18

57 A P A P C H A N T S M V T H P 18

67 M V T H P D F A T Q P Q H V Q 18

74 A T Q P Q H V Q N F L L Y R H 18

115 S S P S N Y V R R E L L R R T 18

149 T A s N P H Ξ A R K V N R L L 18

176 w D F H D S F F N L T L K Q V 18

191 L F L Q W Q Ξ T R C A N A S F 18

208 N G D D D V F A H T D N M V F 18

209 G D D D V F A H T D N M V F Y 18

229 P G R H L F V G Q L I Q N V G 18

268 C G G G G F L L S R F T A A A 18

269 G G G G F L L S R F T A A A L 18

276 S R F T A A A L R R A A H V L 18

80 V Q N F L L Y R H C R H F P L 16

89 C R H F P L L Q D V P P S K C 16

127 R R T W G R E R K V R G L Q L 16

175 Q W D F H D S F F N L T L K Q 16

180 D S F F N L T L K Q V L F L Q 16

192 F L Q W Q E T R C A N A S F V 16

202 N A S F V L N G D D D V F A H 16

211 D D V F A H T D N M V F Y L Q 16

219 N M V F Y L Q D H D P G R H L 16

220 M V F Y L Q D H D P G R H L F 16

231 R H L F V G Q L I Q N V G P I 16

246 R A F W S K Y Y V P E V V T Q 16

249 W S K Y Y V P E V V T Q N E R 16

264 Y P P Y C G G G G F L L S R F 16

270 G G G F L L S R F T A A A L R 16

108 F L L L V I K S S P S N Y V R 15

271 G G F L L s R F T A A A L R R 15

12 T L I L A I G A F T L L L F S 14

14 I L A I G A F T L L L F S L L 14 TABLE XXV - 184P3C10B HLA- DRB1*0401 (DR4D 4) 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

21 T L L L F S L L V S P P T C K 14

24 L F S L L V S P P T c K V Q E 14

26 S L L V S P P T C K V Q E Q P 14

64 N S M V T H P D F A T Q P Q 14

65 T S M V T H P D F A T Q P Q H 14

77 P Q H V Q N F L L Y R H c R H 14

82 N F L L Y R H C R H F P L L Q 14

92 F P L L Q D V P P S K c A Q P 14

106 P V F L L L V I K S S P S N Y 14

143 L L F L V G T A S N P H E A R 14

159 V N R L L E L E A Q T H G D I 14

170 H G D I L Q W D F H D S F F N 14

184 N L T L K Q V L F L Q W Q E T 14

188 K Q V L F L Q W Q Ξ T R C A N 14

210 D D' D V F A H T D N M V F Y L 14

217 T D N M V F Y L Q D H D P G R^' 14

218 D N M V F Y L Q D H D P G R H 14

230 G R H L F V G Q L I Q N V G P 14

255 P Ξ V V T Q N E R Y P P Y C G 14

TABLE XXVI - 184P3C10B HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

106 P V F L L L V I K S S P S N Y 26 127 R R T W G R E R K V R G L Q L 26 105 Q P V F L L L V I K S S P S N 24 219 N M V F Y L Q D H D P G R H L 24

89 C R H F P L L Q D V P P S K C 22 255 P E V V T Q N E R Y P P Y C G 22 269 G G G G F L L S R F T A A A L 21 275 L S R F T A A A L R R A A H V 21

80 V Q N F L L Y R H C R H F P L 20

82 N F L L Y R H C R H F P L Q 20 104 A Q P V F L L L V I K S S P S 20 120 Y V R R E L L R R T W G R E R 20 140 Q R L L F L V G T A S N P H 20 236 G Q L I Q N V G P I R A F W S 20

17 I G A F T L L L F s L L V S P 19

21 T L L L F S L L V s P P T C K 19 107 V F L L L V I K S s P S N Y V 19 180 D S F F N L T L K Q V L F L Q 19 218 D N M V F Y L Q D ,H D P G R H 19 232 H L F V G Q L I Q N V G P I R 19

22 L L L F S L L V S P P T C K V 18

83 F L L Y R H C R H F P L L Q D 18 156 A'R K V N R L L E L E A Q T H 18 239 I Q N V G P I R A F W S K Y Y 18

46 A L A W P T P P T R P A P A P 17 142 R L L F L V G T A S N P H E A 17 179 H D S F F N L T L K Q V L F L 17

47 L A w P T P P T R P A P A P C 16 192 F L Q W Q E T R C A N A S F V 16 ^' 261 N E R Y P P Y C G G G G F L L 16 270 G G G F L L s R F T A A A L R 16

79 H V Q N F L L Y R H C R H F P 15 115 S S P S N Y V R R E L L R R.T 15 119 N Y V R R E L L R R T W G R E 15 TABLE XXVI - 184P3 C10B HLA- DRB1* 1101 15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

153 P H E A R K V N R L L E L E A^' 15

276 S R F T A A A L R R A A H V L 15

280 A A A L R R A A H V L D I F P 15

8 R P N A T L I L A I G A F T L 14

62 H A N T S M V T H P D F A T Q 14 88 H C R H F P L L Q D V P P S K 14 133 E R K V R G L Q L R L L F L V 14 162 L L E L E A Q T H G D I L Q W 14 171 G D I L Q W D F H D S F F N L 14 251 K Y Y V P E V V T Q N E R Y P 14 277 R F T A A A L R R A A H V L D 14 7 R R P N A T L I L A I G A F T 13

19 A F T L L L F s L L V S P P T 13 33 T C K V Q E Q P P A I P E A L 13 37 Q E Q P P A I P Ξ A L A W P T 13 92 F P L L Q D V P P S K C A Q P 13 137 R G L Q L R L L F L V G T A S 13 141 L R L L F L V G T A S N P H E 13 190 V L F L Q W Q E T R C A N A S 13 254 V P E V V T Q N E R Y P P Y C 13 11 A T L I L A I G A F T L L L F 12 20 F T L L L F s L L V S P P T C 12 30 S P P T C K V Q E Q P P A I P 12 41 P A I P E A L A W P T P P T R 12 95 L Q D V P P S K C A Q P V F L 12 117 P S N Y V R R E L L R R T W G 12 138 G L Q L R L L F L V G T A S N 12 159 V N R L L E L E A Q T H G D I 12 160 N R L L E L E A Q T H G D I L 12 170 H G D I L Q W D F H D S F F N 12 184 N L T L K Q V L F L Q W Q E T 12 187 L K Q V L F L Q W Q E T R C A 12 189 Q V L F L Q W Q E T R C A N A 12 204 S F V L N G D D D V F A H T D 12 233 L F V G Q L I Q N .V G P I R A 12 250 S K Y Y V P Ξ V V T Q N E R Y 12 271 G G F L L S R F T A A A L R R 12 283 L R R A A H V L D I F P I D D 12

TABLE XXIII - 184P3G10 HLA- DRB1*0101 15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

265 P I Y W D T I P P D R- V E A A 30 80 A A R F T F L E Q L E S N P T 29

64 S R Y Y E L L K Q R Q A L P I 28 183 L Q E V A S T R G T G A W G V 27 175 R F C W D R L L L Q E V A S T 26 192 T G A W G V L V L D Ξ A Q E R 26 219 D A R L E K L P G D L R V V V 26 240 E P K L R A F W G N P P I V H 26 51 E L A V N P F D G L P F s S R 25 67 Y E L L K Q R Q A L P I. W A A 25 131 V T Q P Y P L A A R S L A L R 25 139 A R S L A L R V A D E M D L T 25 147 A D E M D L T L G H E V G Y S 25 226 P G D L R V V V V T D P A L E 25 230 R V V V V T D P A L E P K L R 25 TABLE XXIII - 184P3G10 HLA- DRB1*0101 15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

280 C Q A V L E L C R K E L P G D 25

11 S V T K Y D L T G C S A F C R 24

29 R T M T S Q P L R L A E E Y 24

36 P L R L A E E Y G P S P G E S 24

66 Y Y E L L K Q R Q A L P I W A 24

82 R F T F L E Q L E S Ν P T G V 24

93 P T G V V L V S G E P G s G K 24 108 S T Q I P Q W C A E F A L A R 24 151 D L T L G H E V G Y S I P Q E 24 252 I V H I P R E P G E R P S P . I 24

6 A G K M V S N T K Y D L T G C 23

83 F T F L E Q E S Ν P T G V V 23

92 N P T G V V L V S σ E P G S G 23 126 K G Q V T V T Q P Y P L A A R 23 128 Q V T V T Q P Y P L A A R S L 23 228 D L R V V V V T D P A L Ξ P K 23

34 S Q P L R L A E E Y G P S P G 22 .

72 Q R Q A L P I W A A R F T F L 22 202 E A Q E R S V A S D S L Q G L 22 180 R L L L Q E V A S T R G T G A 21 186 V A S T R G T G A w G V L V L 21 227 G D L R V V V V T D P A L E P 21

12 V T K Y D L T G C S A F C R S 20

26 S C Q R A T M T S Q P L R L A 20

73 R Q A L P I W A A R F T F L E 20 113 Q W C A E F A L A R G F Q K G 20 171 N T L L R F C W D R L L L Q E 20

94 T G V V L V S G E P G S G K S 19 115 C A E F A L A R G F Q K G Q V 19 132 T Q P Y P L A A R S L A L R V 19 136 P L A A R S L A L R V A D E M 19 159 G Y S I P Q E D C T G P Ν T L 19 176 F C W D R L L L Q E V A S T R 19 195 W G V L V L D E A Q E R S V A 19

40 A E E Y G P S P G E S E L A V 18

49 E S E L A V Ν P F D G L P F S 18

54 V N P F D G L P F S S R Y Y E 18

59 G L P F S S R Y Y E L ^' L K Q R 18

63 S S R Y Y Ξ L L K Q R Q A L P 18

96 V V L V S G E P G S G K S T Q 18 111 I P Q w C A E F A L A R G F Q 18 141 S L R V A D E M D L T L G 18 205 E R S V A S D S L Q G L L Q D 18 237 P A L Ξ P K L R A F W G Ν P P 18 244 R A F W G Ν P P I V H I P R E 18 271 I P P D R V E A A C Q A V L E 18 284 L E L C R K E L P G D V L V F 18

14 K Y D L T G C S A F C R S C Q 17

75 L P I W A A R F T F L E Q L 17

85 F L E Q L E S Ν P T G V V L V 17 177 C W D R L L L Q E V A S T R G 17 188. S T R G T G A W G V L V L D E 17 197 V L V L D E A Q E R S V A S D 17 211 D S L Q G L L Q D A R L Ξ K L 17 214 Q G L L Q D A R L E K L P G D 17 281 Q A V L E L C R K E L P G D V 17

25 R S C Q R A T M T S Q P L R L 16 TABLE XXIII - 184P3G10 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

45 P S P G E S Ξ L A V N P F D G 16

46 S P G E S E L A V N P F D G L 16

69 L L K Q R Q A L P I W A A R F 16

70 L K Q R Q A L P I w A A R F T 16

76 L P I W A A R F T F L E Q L E 16

87 E Q L E S N P T G V V L V S G 16

95 G V V L V S G Ξ P G S G K S T 16

97 V L V S G E P G S G K S T Q I 16

130 T V T Q P Y P L A A R S L A L 16

135 Y P L A A R S L A L R V A D E 16 ^•

143 A L R V A D E M D L T L G H E 16

170 P N T L L R F C W D R L L L Q 16

178 W D R L L L Q E V A S T R G T 16

179 D R L L L Q E V A s T R G T G 16

194 A W G V L V L D E A Q Ξ R S V 16

196 G V L V L D E A Q E R S V A s 16

201 D E A Q E R S V A s D S L Q G 16

207 S V A S D S L Q G L L Q D A R 16

210 S D S L Q G L L Q D A R L Ξ K 16

212 S L Q G L L Q D A R L E K L P 16

222 L E K L P G D L R V V V V T D 16

223 E K L P G D L R V V V V T D P 16

233 V V T D P A L E P K L R A F W 16

243 L R A F W G N P P I V H I P R 16

264 S P I Y w D T I P P D R V E A 16

268 W D T I P P D R V E A A c Q A 16

273 P D R V E A A C Q A V L E L C 16

10 V S V T K Y D L T G C S A F C 15

22 A F C R S C Q R A T M T S Q P 15

42 E Y G P S P G E S E L A V N P 15

100 S G E P G S G K S T Q I P Q W 15

105 S G K S T Q I P Q W C A Ξ F A 15

119 A L A R G F Q K G Q V T V T Q 15

123 G F Q K G Q V T V T Q P Y P L 15

125 Q K G Q V T V T Q P Y P L A A 15

127 G Q V T V T Q P Y P L A A R s 15

152 L T L G H E V G Y s I P Q E D 15

162 I P Q E D C T G P N T L L R F 15

198 L V L D E A Q Ξ R S V A S D S 15

229 L R V V V V T D P A L E P K L 15

248 G N P P I V H I P R E P G E R 15

249 N P P I V H I P R E P G Ξ R P 15

270 T I P P D R V E A A C Q A V L 15

3 T A F A G K M V S V T K Y D L 14

9 M V S V T K Y D L T G C S A F 14

23 F C R S C Q R A T M T S Q P L 14

28 Q R A T M T S Q P L R L A E E 14

39 L A E E Y G P S P G Ξ S E L A 14

53 A V N P F D G L P F S S R Y Y 14

101 G E P G S G K S T Q I P Q W C 14

107 K S T Q I P Q W c A E F A L A 14

133 Q P Y P L A A R s L A L R V A 14

155 G H E V G Y S I P Q E D C T G 14

199 V L D E A Q E R s V A S D S L 14

218 Q D A R L E K L P G D L R V V 14

242 K L R A F W G N P P I V H I P 14

255 I P R E P G E R P S P I Y w D 14

260 G E R P S P I Y w D T I P P D 14 TABLE XXIII - 184P3G10 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

262 R P S P I Y W D T I P P D R V 14 275 R V E A A C Q A V L E L C R K 14

TABLE XXIV - 184P3G10 HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

205 E R S V A S D S L Q G L L Q D 31 213 L Q G L L Q D A R L E K L P G 29 222 L E K L P G D L R V V V V T D 28 149 E M D L T L G H E V G Y S I P 27 280 C Q A V L E L C R K E L P G D 27

6 A G K M V S V T K Y D L T G C 26

141 S A L R V A D E M D L T L G 25

273 P D R V E A A C Q A V L E L C 25

195 W G V L V L D E A Q E R S V A 24

230 R V V V V T D P A L E P K L R 23

268 W D T I P P D R V E A A C Q A 22

51 E L A V N P F D G L P F S S R 20

66 Y Y E L L K Q R Q A L P I w A 20

170 P N T L L R F C w D R L L L Q 20

171 N T L L R F C W D R L L L Q E 20

173 L L R F C W D R L L L Q E V A 20

228 D L R V V V V T D P A L Ξ P K 20

263 P S P I Y W D T I P P D R V E 20

34 S Q P L R L A E E Y G P S P G 19

49 E S E L A V N P F D G L P F S 19

65 R Y Y E L L K Q R Q A L P I W 19

82 R F T F L E Q L E S N P T G V 19

94 T G V V L V S G E P G S G K S 19

126 K G Q V T V T Q P Y P L A A R 19

143 A L R V A D E M D L T L G H E 19

194 A W G V L V L D E A Q E R S V 19

214 Q G L L Q D A R L E K L P G D 19

236 D P A L E P K L R A F W G N P 19

59 G L P F S S R Y Y E L L K Q R 18

75 A L P I w A A R F T F L E Q L 18

196 G V L V L D E A Q E R S V A S 18

210 S D S L Q G L L Q D A R L E K 18

249 N P P I V H I P R E P G E R P 18

281 Q A V L Ξ L c R K E L P G D V 18

57 F D G L P F s S R Y Y E L L K 17

80 A A R F T F L E Q L E S N P T 17

197 V L V L D E A Q E R S V A S D 17

232 V V V T D P A L E P K L R A F 17

41 E E Y G P S P G E S E L A V N 16

117 E F A L A R G F Q K G Q V T V 16

163 P Q E D C T G P N T L L R F C 16

172 T L L R F C W D R L L L Q E V 16

206 R S V A S D S L Q G L L Q D A 16

250 P P I V H I P R E P G E R P S 16

20 C S A F C R s C Q R A T M T S 15

74 Q A L P I W A A R F T F L E Q 15

109 T Q I P Q W c A E F A L A R G 15

178 W D R L L L Q E V A S T R G T 15

TABLE XXV - 184P3G10 HLA- DRB1*0401 (DR4DW4) 15 - mers Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

S R Y Y E L L K Q R Q A L P I 28

A A R F T F L E Q L E S N P T 28

T Q P Y P L A A R S L A L R V 28

F T F L E Q L E S N P T G V V 26

G V L V L D E A Q E R S V A S 26

L Q G L L Q D A R L E K L G 26

L E K L P G D L R V V V V T D 26

P D R V E A A C Q A V L E L C 26

C Q A V L E L c R K E L P G D 26

C S A F C R S c Q R A T M T S 22

S S R Y Y E L L K Q R Q A P 22

C A Ξ F A L A R G F Q K G Q V 22

L L R F C W D R L L L Q E V A 22

R F C W D R L L L Q E V A S T 22

T G A W G V L V L D E A Q E R 22

P I Y W D T I P P D R V E A A 22

A G K M V S V T K Y D L G C 20

A L P I W A A R F T F L E Q L 20

L E Q L E S N P T G V V L V S 20

V V L V S G Ξ P G S G K S T Q 20

E F A L A R G F Q K G Q V T V 20

S L A L R V A D E M D L T L G 20

A L R V A D Ξ M D L T L G H E 20

D L T L G H E V G Y S I P Q E 20

P N T L L R F C W D R L L Q 20

R L L L Q E V A S T R G T G A 20

V L V L D E A Q E R S V A S D 20

E R S V A S D S L Q G L L Q D 20

S D S L Q G L L Q D A R L E K 20

Q G L L Q D A R L E K L P G D 20

D L R V V V V T D P A L E P K 20

N P P I V H I P R E P G E R P 20

W D T I P P D R V E A A C Q A 20

V S V T K Y D L T G C S A F C 18

L T G c s A F C R S C Q R T 18

S G K s T Q I P Q W C A E F A 18

W C A E F A L A R G F Q K G Q 18

Q P Y P L A A R S L A L R V A 18

Y S I P Q E D C T G P N T L L 18

F C W D R L L L Q E V A S T R 18

L L L Q E V A S T R G T G A W 18

V L D E A Q E R s V A S D S 18

E A Q E R S V A s D S L Q G L 18

G D L R V V V V T D P A L E P 18

V N P F D G L P F S S R Y Y E 16

I P Q W C A E F A L A R G F Q 16

A R G F Q K G Q V T V T Q P Y 16

E V G Y S I P Q E D C T G P N 16 A F W G N P P I V H I P R E 16

G K M V S V T K Y D L T G C S 14

R A T M T S Q P L R L A E E Y 14

S Q P L R L A E E Y G P S P G 14

P L R L A E Ξ Y G P S P G E S 14

E S Ξ L A V N P F D G L P F S 14

E L A V N P F D G L P F S S R 14

F D G L P F S S R Y Y E L L K 14

Y Y E L L K Q R Q A L P I W A 14

R Q A L P I W A A R F T F L E 14

P T G V V L V S G E P G S G K 14

T G V V L V S G E P G S G K S 14 TABLE XXV - 184P3G10 HLA- DRB1*0401 (DR4DW4) 15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

108 S T Q I P Q W c A E F A L A R 14 126 K G Q V T V T Q P Y P L A A R 14 128 Q V T V T Q P Y P L A A R S L 14 147 A D Ξ M D L T L G H E V G Y S 14 159 G Y S I P Q E D c T G P N T L 14 171 N T L L R F c W D R L L L Q E 14 178 W D R L L L Q E V A S T R G T 14 179 D R L L L Q E V A S T R G T G 14 183 L Q E V A S T R G T G A W G V 14 194 A W G V L V L D E A Q E R S V 14 195 W G V L V L D E A Q E R S V A 14 219 D A R L E K L P G D L R V V V 14 226 P G D L R V V V V T D P A L E 14 230 ^°R V V V V T D P A L E P K L R 14 231 V V V V T D P A L E P K L R A 14 240 E P K L R A F W G N P P I V H 14 252 I V H I P R E P G E R P s P I 14 263 P S P I Y W D T I P P D R V E 14

TABLE XXVI - 184P3G10 HLA- DRB1*1101 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score

63 s S R Y Y E L L K Q R Q A L P 26

249 N P P I V H I P R E P G E R P 26

132 T Q P Y P L A A R S L A L R V 25

192 T G A W G V L V L D E A Q E R 25

80 A A R F T F L E Q L Ξ S N P T 24

64 S R Y Y E L L K Q R Q A L P I 23

148 D E M D L T L G H E V G Y S I 21

226 P G D L R V V V V T D P A L E 21

281 Q A V L E L C R K E L P G D V 21

6 A G K M V S V T K Y D L T G C 20

180 R L L L Q E V A S T R G T G A 20

216 L L Q D A R L Ξ K L P G D L R 20

280 c Q A V L E L C R K E L P G D 20

93 P T G V V L V S G E P G S G K 18

265 P I Y W D T I P P D R V Ξ A A 18

29 R A T M T S Q P L R L A Ξ E Y 16

54 V N P F D G L P F S S R Y Y E 16

82 R F T F L E Q L E S N P T G V 16

90 E S N P T G V V L V S G E P G 16

175 R F C W D R L L L Q E V A S T 16

234 V T D P A L E P K L R A F W G 16

246 F W G N P P I V H I P' R^'E P G 16

92 N P T G V V L V S G E P G S G 15

114 W c A E F A L A R G F Q K G Q 15

198 L V L D E A Q E R S V A S D S 15

222 L E K L P G D L R V V V V T D 15

227 G D L R-V V V V T D P A L Ξ P 15

57 F D G L P F S S R Y Y E L L K 14

105 S G K S T Q I P Q W C A E F A 14

128 Q V T V T Q P Y P L A A R S L 14

136 P L A A R S L A L R V A D E M 14

167 c T G P N T L L R F C W D R L 14

182^' L L Q E V A S T R G T G A W G 14

196 G V L V L D E A Q E R S V A S 14

213 L Q G L L Q D A R L E K L P G 14

236 D P A L E P K L R A F W G N P 14 TABLE XXVI - 184P3G10 HLA-

DRB1*1101 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score

252 I V H I P R E P G E R P S P I 14

254 H I P R E P G E R P S P I Y W 14

267 Y w D T I P P D R V E A A C Q 14

66 Y Y E L L K Q R Q A L P I W A 13

94 T G V V L V S G E P G S G K S 13

144 L R V A D E M D L T L G H E V 13

152 L T L G H E V G Y S I P Q E D 13

176 F C W D R L L L Q E V A S T R 13

194 A W G V L V L D E A Q E R S V 13

219 D A R L E K L P G D L R V V V 13

228 D L R V V V V T D P A L E P K 13

11 S V T K Y D L T G C S A F C R 12

33 T S Q P L R L A E E Y G P S P 12

34 s Q P L R L A E E Y G P S P G 12

36 P L R L A E E Y G P S P G E S 12

48 G E S E L A V N P F D G L P F 12

70 L K Q R Q A L P I W A A R F T 12

83 F T F L E Q L E s N P T G V V 12

95 G V V L V S G E P G S G K S T 12

108 S T Q I P Q W C A E F A L A R 12

123 G F Q K G Q V T V T Q P Y P L 12

125 Q K G Q V T V T Q P Y P L A A 12

159 G Y S I P Q E D C T G P N T L 12

177 C W D R L L L Q E V A S T R G 12

183 L Q E V A S T R G T G A W G V 12

210 S D S L Q G L L Q D A R L E K 12

230 R V V V V T D P A L E P K L R 12

237 P A L E P K L R A F W G N P P 12

240 E P K L R A F W G N P P I V H 12

268 W D T I P P D R V E A A C Q A 12

TABLE XXIII - 185P2C9 vl HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

104 R L Q I S E L s G K V L K L Q 33 908 Q S G L R V L H S P P A V R R 32 478 L G E L G V Q G G H Q A D G P 31 696 V R P F P H Q G S L R M P R P 31 902 S H V L T E Q S G L R V L H s 31 181 E E M F E K T S G F G S G K P 30 917 P P A V R R V D S I T A A G G 30 932 E G P F P T s R A R G S P G D 29 39 H L Q F V E E E A E L L R R S 28 358 R Ξ S L R L R A A R E L H R R 28 399 A L R W K Ξ L E M H S L A L Q 28 794 F Q R L M D I S P F L P ^■E K G 28 923 V D S I T A A G G E G P F P T 28

1165 S S L F N I I D H S P V V Q D 28

68 L S K F K F E P P R E P G w L 27

472 P E T L S R L G E L G V Q G G 27

1065 H K C L T P K A G G G A T P V 27 246. L R G G A P L P G P G L Q G E 26 327 I G N L G K E L G P D L Q S R 26 429 Q Q E L R S L K Q N I F L F Y 26 711 V A M W P c A D A D S I P F E 26 791 M S E F Q R L M D I S P F L P 26 800 I S P F L P Ξ K G L P s T S S 26 TABLE XXIII - 185P2C9 vl HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1007 E V A F S V R N A I C S G P G 26

1049 G L Q T E A L R G S G V T S S 26

1244 G Y T L T E N V A R I L N K K 26

1264 L K E E R R Q A A H G P P G L 26

136 H L G L R A P S P R D S D A E 25

394 E E H L Y A L R W K E L E M H 25

517 D H S L R L Q T A D R G Q P H 25

560 R A R L R L Q Q Q Y A S D K A 25

702 Q G S L R M P R P V A M W P C 25

873 D S S W Y L T T S V T M T T D 25

956 L S R W P C T s P R H S R D Y 25

1080 S S P S R s L R S R Q V A P A 25

1092 A P A I E K V Q A K F E R T C 25

79 P G W L G E G A S P G A G G G 24

112 G K V L K L Q H E N H A L L S 24

130 R C D L A A H L G L R A P S P 24

133 L A A H L G L R A P S P R D S 24

187 T S G F G S G K P s E A S E P 24

240 F L H D A G L R G G A P L P G 24

268 Q Q E P Q L L G T I N A K M K 24

292 L E Q V N R I G D G L S P L P 24

302 L S P L P H L T E s S S F L S 24

311 S S S F L S T V T S V S R D S 24

424 Ξ K N L M Q Q E L R S L K Q N 24

508 S P H S R V Q I G D H S L R L 24

658 R Q E W E R Q K K E F L W R I 24

669 L W R I E Q L Q K E N S P R R 24

704 S L R M P R P V A M W P c A D 24

740 S E N L Y L D A L S L D D E 24

777 N H K G N L Q R A V S V S S M 24

803 F P E K G L P S T S s K E D 24

920 V R R V D S I T A A G G E G P 24

979 D S P L C T s L G F A S P L H 24

994 S L E M S K N L S D D M K Ξ V 24

1236 K Q D L S A P P G Y T L T E N 24

1248 T E N V A R I L N K K L L E H 24

100 L K S A R L Q I S E L S G K V 23

271 P Q L L G T I N A K M K A F K 23

305 L P H L T E s S S F L S T V T 23

309 T E S S S F L S T V T S V S R 23

315 L S T V T S V S R D S P I G N 23

339 Q S R L K E Q L E W Q L G P A 23

438 N I F L F Y V K L R W L L K H 23

595 E N K L G E L G S S A E S K G 23

983 C T S L G F A S P L H S L E M 23

1032 T G S R T M G T Q T V Q T I 23

1044 Q T I S V G L Q T E A L R G s 23

1157 H T T I N D G L s S L F N I I 23

1176 V V Q D P F Q K G L R A G s R 23

1232 G P V K Q D L S A P P G Y T 23

94 A P L Q E Ξ L K S A R L Q I S 22

122 H A L L S N I Q R C D L A A H 22

227 E R T V E R L I T D T D S F L 22

295 V N R I G D G L s P L P H L T 22

343 K E Q L Ξ W Q L G P A R G D E 22

345 Q L E W Q L G P A R G D E R E 22

379 S H G L G G Q T C F S L E M E 22

532 K Q V V E N Q Q L F S A F K A 22 TABLE XXIII - 185P2C9 vl HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

562 R L R L Q Q Q Y A S D K A A W 22

592 E K T E N K L G E L G S S A E 22

619 Q K L L A D S H S L V M D L R 22

685 G S F L C D Q K D G N V R P F 22

779 K G N L Q R A V S V S S M S E 22

816 E D V T P P L S P D D L K Y I 22

894 H C Q K Q P L R S H V L T E Q 22

952 P E M L S R W P C T S P R H 22

986 L G F A S P L H S L E M S K N 22

1035 S R T M G T Q T V Q T I S V G 22

1084 R S L R S R Q V A P A I E K V 22

1164 L S S L F N I I D H S P V V Q 22

1178 Q D P F Q K G L R A G s R S R 22

1249 E V A R I L N K K L L E H A 22

57 I E D H N R Q L T H E L S K F 1

263 Q G E G D Q Q E P Q L L G T I 21 281 M K A F K K E L Q A F L E Q V 21

293 E Q V N R I G D G L S P L P H 21

407 M H S L A L Q N T L H E R T W 21

440 F L F Y V K L R W L L K H w R 21

830 I E E F N K S W D Y T P N R G 21

878 L T T S V T M T T D T M T S P 21

949 G G P P E P M L s R W P C T s 21

15 R D H A P s I P T S P F G D s 20

312 S S F L S T V T s V S R D S P 20

401 R W K E L E M H s L A L Q N T 20

631 D L R W Q I H H s Ξ K N W N R 20

693 D G N V R P F P H Q G S L R M 20

798 M D I S P F L P E K G L P S T 20

812 T S S K E D V T P P L S P D D 20

839 Y T P N R G H N G G G P D L W 20

898 Q P L R S H V L T E Q s G L R 20

967 S R D Y V E G A R R P L D S P 20

1009 A F S V R N A I C S G P G E L 20

1037 T M G T Q T V Q T I S V G L Q ^■20

1052 T E A L R G S G V T S S P H K 20

1155 P V H T T I N D G L S S L F N 20

78 E P G W L G E G A S P G A G G 19

215 Y L V T L K H E A Q R L E R T 19

235 T D T D S F L H D A G L R G G 19

291 F L E Q V N R I G D G L S P L 19

475 L S R L G E L G V Q G G H Q A 19

541 F S A F K A L L E D F R A E L 19

552 R A E L R E D E R A R L R L Q 19

577 D V Ξ W A V L K C R L E Q L Ξ 19

640 E K N W N R E K V E L L D R L 19

721 S I P F E D R P L S K L K E S 19

836 S W D Y T P N R G H N G G G P 19

1047 S V G L Q T E A L R G S G V T 19

1110 K Y G S P K L Q R K P L P K A 19

1174 S P V V Q D P F Q K G L R A G 19

1213 R S P s P I G V G S E M C R E 19

1252 A R I L N K K L L E H A L K E 19

1253 R I L N K K L L E H A L K E E 19

58 E D H N R Q L T H E L S K F K 18

65 T H E L S K F K F E P P R E P 18

75 P P R E P G W L G E G A S P G 18

77 R E P G W L G E G A S P G A G 18 H H 1 t-- r-^l r r-ⁱ t-- t-- i-r -

_^i en rf* rf* H to to to to to vo ω vo vo co ι ^ cn cn cn ιπ ιn iπ ιπ ιπ ιπ ιπ ιπ ιf rf* rf* ιf* rf rf* to to to to t H H H H H H H cn to rf* LO vo co t H o o oo K w S S S S S S ∞ j en rf* LO LO VD CD Cn rf* L _ rf* _o tO H O VO Cθ σι LO tO _ H co TJ o tO rf* H VO rf* tO _0 -~_ Ul l C0 l ~J. H VO ∞ O ^ cn ^ιo ∞ H VD CD H ^ H l - ιf* ^cn "ιπ cn ιπ vo rf* vo ιo σι _o co tfi H H H Lo vD iπ H -J CD tθ H θ iπ to H cn L H 01

a a a <! a a 3 3 a a lO.KD B KD rt cn a Ω Ω a cn a B a cn a a Ω a a a i CO KD Ω a a a a rt a α a a W B Ω ω Ω a o a B O o a a Ω Ω a a a Ω a rt a CO cn a cn TI < a a a . M B Ω S! > KD κD B a Ω B < a a a α rt cn Ω Ώ a co α a a Ω a to a a B B 3 a cn a a a rt Ω a Ω Ω cn a rt Ω α a a a Ω rt a a cn X a a ;> ιo a a a D <! Ω a a o ω <; B a rt a a a a Ω O a rt a a a 3 a a cn Ω Ω a < a a a <! o a Ω o o 3 a a a o a Ω a <: s a a KD a a a a a a a rt Ω cn tr co <: a rf* ω

Ω a B rt Ω a Ω a a a rt a a ω a a artsartκ;rtrt ω < a K cn a a o ;> co KD o Ω a a Ω a a a a a Ω a a a a H

*

Si a a a KD Ω α Ω cn a a a cn Ω co a cn n < a rt a a 3 a a co KD a < ω a a n Ω a Si a a a a < a a a o a Ω a a O cn H a a a a •O O O rt Ω a a cn Ω co a Ω Ω cn a a a B Ω a a a o a a > a Ω a α a a a α X rt a a a a o rt a cn Ω a a α KD a O

Ω H

ON Ω cn a a a B IO a a cn Ω KD Ω Ω a < a Ω Ω Ω rt a a a a a a a ; a a a a D B a a a Ω > rt a a a n a Ω a KD 5 oo o

B a a a Ω a B a Ω Ω a Ω a a i < > Ω a > 3 a > a 3 a a a α cn B a iso > Ω > Ω a a Ω Ω o a Ω a a Ω a Ω vo

Ω a a α B a a rt < a cn α a rt > a a cn a a rt a cn s; cn o a a a < le' a a a Ω α B a Si < > a rt a a cn a a a > α B Ω o a cn O Ω a Ω a Ω Ω cn a Ω a a a a rt Ω cn a cn a B cn B α KD X Ω rt KD a Si a a a Ω Ω a -3 a a a a α a a B a Ώ 5=- X a a Ω 3 ro

^' cn a co a a Ω a < < a a co a > a a < a a a a a B a H cn a a a a α B : a a a a a a a Ω cn o a cn a a a a a B a a Ω a 3 a a Ω ω rt O cn < rt a a a a 3 a B Ω a a a a a Ω < a o a a α B a a a Ω a iO cn Ω a Ω a ω

Ω T) B cn a Ω a KD 3 co a a rt a B cn n a a a α a a a co a <. a a a a a B a a a a Ω X Si Ω a a B a i Ω a a a a a rf_* a a cn α a a o Ω a a a rt a a a cn a o ω Ω a cn 3 a a a a S a a B Ω α a a a TJ a a rt to Ω a a a a KD

H H H H H H H H 1 -J -0 co co oo co co fl)

TABLE XXIII - 185P2C9 l HLA- DRB1*0101 15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

84 E G A S P G A G G G A P L Q E 17

85 G A S P G A G G G A P L Q E E 17

91 G G G A P L Q E E L K S A R L 17

97 Q E E L K S A R L Q I S E L S 17 152 D A G K K E S D G E E S R L P 17 179 D S E E M F E K T S G F G S G 17 213 S E Y L V T L K H E A Q R L E 17 223 A Q R L E R T V E R It I T D T 17 229 T V E R L I T D T D S F L H D 17 242 H D A G L R G G A P L P G P G 17 267 D Q Q E P Q L L G T I N A K M 17 270 E P Q L L G T I N A K M K A F 17 275 G T I N A K M K A F K K E L Q 17 282 K A F K K E L Q A F L E Q V N 17 288 L Q A F L E Q V N R I G D G L 17 323 R D S P I G N L G K E L G P D 17 324 D S P I G N L G K E L G P D L 17 355 G D E R E S L R L R A A R E L 17 371 R R A D G D T G S H G L G G Q 17 376 D T G S H G L G G Q T C F S L 17 421 W S D E K N L M Q Q Ξ L R S L 17 448 W L L K H W R Q G K Q M E E E 17 455 Q G K Q M E E E G E E F T E G 17 497 D R G C G F P V G E H S P H S 17 523 Q T A D R G Q P H K Q V V E N 17 531 H K Q V V E N Q Q L F S A F K 17 587 L E Q L E E K T E N K L G E L 17 611 L K K E R E V H Q K L L A D S 17 645 R E K V E L L D R L D R D R Q 17 677 K E N S P R R G G S F L C D Q 17 708 P R P V A M W P C A D A D S I 17 719 D S I P F E D R P L S K L K 17 729 L S K L K E S D R C S A S E N 17 792 S E F Q R L M D I s P F L P E 17 819 T P P L S P D D L K Y I E E F 17 841 P N R G H N G G G P D L W A D 17 856 R T E V G R A G H E D S T E P 17 925 S I A A G G E G P F P T S R 17 953 E P M L S R W P C T S P R H S 17 975 R R P L D S P L C T S L G F A 17 977 P L D S P L C T S L G F A S P 17 981 P L C T s L G F A S P L H S L 17 1005 M K Ξ V A F S V R N A I C S G 17 1017 C S G P G E L Q V K D M A C Q 17 1039 G T Q T V Q T I S V G L Q T E 17 1043 V Q T I S V G L Q T E A L R G 17 1061 T S S P H K C L T P K A G G G 17 1113 S P K L Q R K P L P K A D Q P 17 1116 L Q R K P L P K A D Q P N N R 17 1124 A D Q P N N R T S P G M A Q K 17 1132 S P G M A Q K G Y S Ξ S A W A 17 1142 E S A W A R S T T T R E S P V 17 1143 S A W A R S T T T R E S P V H 17 1166 S L F N I I D H S P V V Q D P 17 1179^' D P F Q K G L R A G S R S R S 17 1180 P F Q K G L R A G S R S R S A 17 1199 E L G P G Q E T G T N S R G R 17 1221 G S E M C R E E G G E G T P V 17 TABLE XXIII - 185P2C9 vl HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1231 E G T P V K Q D S A P P G Y 17 1261 E H A L K E E R R Q A A H G P 17 1267 E R R Q A A H G P P G L H S D 17 10 R E G P G R D H A P S I P T S 16 11 E G P G R D H A P s I P T s P 16 18 P S I P T S P F G D S L E S 16 25 P F G D S L E S S T E L R R H 16 37 R R H L Q F V E E E A E L L R 16 46 E A E L L R R S I S E I E D H 16 47 A Ξ L L R R S I S E I E D H N 16 61 N R Q L T H E L S K F K F E P 16 99 E L K S A R L Q I S E L S G K 16 114 V L K L Q H E N H A L L S N I 16 115 L K L Q H Ξ N H A L L S N I Q 16 127 N I Q R C D L A A H L G L R A 16 155 K K E S D G E E S R L P Q P K 16 167 Q P K R E G P V G G E s D S E 16 184 F E K T S G F G S G K P S E A 16 230 V E R L I T D T D S F L H D A 16 243 D G L R G G A P L P G P G L 16 251 P L P G P G L Q G E E E Q G E 16 289 Q A F L E Q V N R I G D G L S 16 299 G D G L S P L P H L T E S S S 16 303 S P L P H L T Ξ S S S F L S T 16 314 F L S T V T S V S R D S P I G 16 318 V T S V S R D S P I G N L G K 16 328 G N L G K E L G P D L Q S R L 16 331 G K E L G P D L Q S R L K E Q 16 335 G P D L Q S R L K E Q L E W Q 16 347 E W Q L G P A R G D E R Ξ S L 16 357 E R E S L R L R A A R E L H R 16 389 S L E M E E E H L Y A L R W K 16 391 E M E E E H L Y A L R W K Ξ L 16 404 E L E M H S L A L Q N T L H E 16 431 E L R S L K Q N I F L F Y V K 16 493 D H D S D R G c G F P V G E H 16 509 P H S R V Q I G D H S L R L Q 16 514 Q I G D H S L R L Q T A D R G 16 519 S L R L Q T A D R σ Q P H K Q 16 537 N Q Q L F S A F K A L L E D F 16 544 F K A L L E D F R A E L R E D 16 545 K A L L Ξ D F R A Ξ L R E D E 16 559 E R A R L R L Q Q Q Y A S D K 16 567 Q Q Y A S D K A A w D V E W A 16 594 T E N K L G E L G S S A E S K 16 614 E R E V H Q K L L A D S H s L 16 618 H Q K L L A D S H S L V M D L 16 626 H S L V M D L R W Q I H H S E 16 629 V M D L R W Q I H H s E K N W 16 648 V E L L D R L D R D R Q E W E 16 667 E F L W R I E Q L Q K E N S P 16 672 I E Q L Q K E N S P R R G G S 16 676 Q K E N S P R R G G S F L C D 16 699 F P H Q G S L R M P R P V A M 16 700 P H Q G S L R M P R P V A M W 16 710 P V A M W P c A D A D S I P F 16 733 K E S D R c S A S E N L Y L D 16 738 C S A S E N L Y L D A L s L D 16 TABLE XXIII - 185P2C9 l HLA- DRB1* 0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

742 E N L Y L D A L S L D D E P E 16 743 N L Y L D A L S L D D E P E E 16 748 A L S L D D E P E E P P A H R 16 752 D D E P E E P P A H R P E R E 16 776 E N H K G N L Q R A V S V S s 16 780 G N L Q R A V s V S S M S E F 16 783 Q R A V S V S s M S E F Q R L 16 802 P F L P E K G L P S T S S K E 16 824 P D D L K Y I Ξ E F N K s W D 16 846 G G G P D L W A D R T E V G 16 " 905 L T Ξ Q S G L R V L H S P P A 16 909 S G L R V L H S P P A V R R V 16 911 L R V L H S P P A V R R V D S 16 941 R G S P G D T K G G P P E P M 16 972 E G A R R P L D S P L C T S L 16 990 S P L H S L E M S K N L s D D 16 998 S K N L S D D M K E V A F S V 16 1013 R N I C S G P G E L Q V K D 16 1024 Q V K D M A C Q T N G S R T M 16 1027 D M A C Q T N G S R T M G T Q 16 1051 Q T E A L R G S G V T S S P H 16 1062 S S P H K C L T P K A G G G A 16 1075 G A T P V S S P S R S L R S R 16 1097 K V Q A K F E R T C C S P K Y 16 llll Y G S P K L Q R K P L P K A D 16 1133 P G M A Q K G Y S E S A W A R 16 1161 N D G L s S L F N I I D H S P 16 1207 G T N S R G R S P s P I G V σ 16 1233 T P V K Q D L s A P P G Y T L 16 1241 A P P G Y T L T E N V A R I L 16 1269 R Q A A H G P P G L H S D s H 16 1278 L H S D S H S L G D T A Ξ P G 16

TABLE XXIV - 185P2C9 vl HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

544 F K A L L £ D F R A E L R E P 38

552 R A E L R E D E R A R L R L Q 38

331 G K E L G P D L Q S R L K E Q 36

230 V E R L I T D T D S F L H D A 29

625 S H S L V M D L R W Q I H H S 29

429 Q Q E L R S L K Q N I F L F Y 28

339 Q S R L K E Q L E W Q L G P A 27

387 C F S L E M E E E H L Y A L R 27

537 N Q Q L F S A F K A L L E D F 27

587 L E Q L E E K T E N K L G E L 27

746 L D A L S L D D E P E E P P A 27

53 S I S E I E D H N R Q L T H E 26

173 P V G G E S D S E E M F E K T 26

335 G P D L Q S R L K E Q L E w Q 26

350 L G P A R G D E R E S L R L R 26

651 L D R L D R D R Q E W E R Q K 26

684 G G S F L-C D Q K D G N V R P 26

998 S K N L S D D M K E V A F S V 26

1248 T E N V A R I L N K K L L E H 26

281 M K A F K K E L Q A F L E Q V 25

122 H A L L S N I Q R C D L A A H 24 TABLE XXIV - 185P2C9 vl HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

439 I F L F Y V K L R W L L K H W 24

618 H Q K L L A D S H S L V M D L 24

1153 E S P V H T T I N D G L S S L 24

1173 H S P V V Q D P F Q K G L R A 24

1174 S P V V Q D P F Q K G L R A G 24

1232 G T P V K Q D L s A P P G Y T 23

1275 P P G L H S D S H S L G D T A 23

741 S E N L Y L D A L S L D D E P 22

126 S N I Q R C D L A A H L G L R 21

237 T D S F L H D A G L R G G A P 21

318 V T S V S R D S P I G N L G K 21

793 E F Q R L M D I S P F L P E K 21

819 T P P L S P D D L K Y I E E F 21

881 S V T M T T D T M T S P E H C 21

1057 G S G V T S S P H K C L T P K 21

39 H L Q F V E E E A E L L R R s 20

104 R L Q I S E L S G K V L K L Q 20

114 V L K L Q H E N H A L L S N I 20

204 T E L L K A R E D S E Y L V T 20

206 L L K A R Ξ D S E Y L V T L K 20

270 E P Q L L G T I N A K M K A F 20

294 Q V N R I G D G L S P L P H L 20

358 R E S L R L R A A R Ξ L H R R 20

389 S L E M E Ξ E H L Y A L R W K 20

417 H E R T W S D E K N L M Q Q E 20

432 L R S L K Q N I F L F Y V K L 20

511 S R V Q I G D H S L R L Q T A 20

579 E W A V L K C R L E Q L E E K 20

580 W A V L K C R L E Q L E E K T 20

811 S S S K E D V T P P L S P D 20

849 G P D L w A D R T E V G R A G 20

860 G R A G H E D S T E P F P D S 20

867 S T E P F P D S S W Y L T T S 20

975 R R P L D S P L C T s L G F A 20

1156 V H T T I N D G L S s L F N I 20

1157 H T T I N D G L S S L F N I I 20

1244 G Y T L T E N V A R I L N K K 20

1261 E H A L K E E R R Q A A H G P 20

40 L Q F V Ξ E E A E L L R R S I 19

93 G A P L Q E E L K S A R L Q I 19

112 G K V L K L Q H E N H A L L s 19

121 N H A L L S N I Q R C D L A A 19

214 E Y L V T L K H E A Q R L E R 19

278 N A K M K A F K K E L Q A F L 19

315 L S T V T S V S R D s P I G N 19

327 I G N L G K E L G P D L Q S R 19

424 E K N L M Q Q E L R S L K Q N 19

438 N I F L F Y V K L R W L L K H 19

446 L R W L L K H w R Q G K Q M E 19

491 G P D H D s D R G C G F P V G 19

531 H K Q V V E N Q Q L F S A F K 19

560 R A R L R L Q Q Q Y A S D K A 19

768 R N R L P E E E Ξ N H K G N L 19

783 Q R A V S V S S M S E F Q R L 19

824 P D D L K Y I E E F N K S W D 19

992 L H S L E M s K N L S D D M K 19

1045 T I S V G L Q T E A L R G S G 19

1076 A T P V S S P s R S L R S R Q 19 TABLE XXIV - 185P2C9 l HLA- DRB1 * 0301 (DR17 ) 15 - mers

Pos • 0 Seq. ID

1 2 3 4 5 6 7 8 9 1 2 3 4 5 score Num.

1236 K Q D L S A P P G Y T L T E N 19

27 G D S L E S S T E L R R H L Q 18

33 S T E L R R H L Q F V E E E A 18

37 R R H L Q F V E E E A E L L R 18

54 I S E I E D H N R Q L T H E L 18

113 K V L K L Q H E N H A L L S N 18

153 A G K K E S D G E E S R L P Q 18

203 P T E L L K A R E D S E Y L V 18

216 L V T L K H E A Q R L E R T V 18

254 G P G L Q G E E E Q G E G D Q 18

285 K K E L Q A F L E Q V N R I G 18

324 D s P I G N L G K E L G P D L 18

360 S L R L R A A R E L H R R A D 18

379 S H G L G G Q T C F S L E M E 18

386 T C F S L E M E E E H L Y A L 18

394 E E H L Y A L R W K E L E M H^" 18

425 K N L M Q Q E L R S L K Q N I 18

443 Y V K L R W L L K H W R Q G K 18

447 R W L L K H W R Q G K Q M E E 18

456 G K Q M E E E G E E F T E G E 18

472 P E T L S R L G E L G V Q G G 18

558 D E R A R L R L Q Q Q Y A S D 18

598 L G E L G S S A E S K G A L K 18

608 K G A L K K E R E V H Q K L L 18

633 R W Q I H H S E K N W N R E K 18

645 R E K V E L L D R L D R D R Q 18

665 K K E F L W R I E Q L Q K E N 18

666 K E F L W R I Ξ Q L Q K E N S 18

687 F L C D Q K D G N V R P F P H 18

721 S I P F E D R P L S K L K E S 18

729 L s K L K E S D R C s A S Ξ N 18

734 E S D R C S A S E N L Y L D A 18

785 A V S V s S M S E F Q R L M D 18

788 V S S M s E F Q R L M D I S P 18

1005 M K E V A F S V R N A I c S G 18

1091 V A P A I E K V Q A K F E R T 18

1182 Q K G L R A G s R S R S A E P 18

TABLE XXV - 185P2C9 vl HLA- DRB1* 0401 (DR4D 4 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

281 M K A F K K E L Q A F L E Q V 28

418 E R T W S D E K N L M Q Q E L 28 1165 S S L F N I I D H S P V V Q D 28 61 N R Q L T H E L S K F K F E P 26 114 V^' L K L Q H E N H A L L S N I 26 122 H A L L S N I Q R C D L A A H 26 213 S E Y L V T L K H E A Q R L E 26 331 G K E L G P D L Q S R L K E Q 26 425 K N L M Q Q E L R S L K Q N I 26 544 F K A L L E D F R A Ξ L R E D 26 552 R A E L R E D E R A R L R L Q 26 618 H Q K L L A D S H S L V M D L 26 625 S H S L V M D L R W Q I H H S 26 648 V E L L D R L D R D R Q E W E 26 666 K E F L W R I E Q L Q K E N.S 26 693 D G N V R P F P H Q G S L R M 26 TABLE XXV - 185P2C9 vl HLA- DRB1*0401 (DR4DW4 ) 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score •^J=v_,. -

Num.

719 A D S I P F E D R P L S K L K 26

779 K G N L Q R A V S V S S M S E 26

856 R T Ξ V G R A G H E D S T E P 26

908 Q S G L R V L H S P P A V R R 26

917 P P A V R R V D S I T A A G G 26

1118 R K P L P K A D Q P N N R T S 26

1248 T E N V A R I L N K K L L E H 26

1261 E H A L K E E R R Q A A H G P 26

39 H L Q F V E E E A E L L R R S 22

311 S S S F L S T V T S V S R D S 22

399 A L R W K E L E M H S L A L Q 22

440 F L F Y V K L R W L L K H W R 22

463 G E E F T E G E H P E T L S R 22

548 L E D F R A E L R E D E R A R 22

573 K A A W D V E W A V L K C R L 22

631 D L R W Q I H H S E K N W N R 22

640 E K N W N R E K V E L L D R L 22

684 G G S F L C D Q K D G N V R P 22

696 V R P F P H Q G S L R M P R P 22

711 V A M W P C A D A D S I P F E 22

742 E N L Y L D A L S L D D E P E 22

800 I S P F L P E K G L P S T S S 22

826 D L K Y I E Ξ F N K S W D Y T 22

873 D S S W Y L T T S V T M T T D 22

967 S R D Y V E G A R R P L D S P 22

1178 Q D P F Q K G L R A G S R S R 22

54 I S E I E D H N R Q L T H E L 20

79 P G W L G E G A S P G A G G G 20

97 Q E E L K S A R L Q I S E L S 20

104 R L Q I s E L S G K V L K L Q 20

111 S G K V L K L Q H E N H A L L 20

216 L V T L K H E A Q R L E R T V 20

227 E R T V E R L I T D T D S F L 20

230 V E R L I T D T D S F L H D A 20

270 E P Q L L G T I N A K M K A F 20

271 P Q L L G T I N A K M K A F K 20

278 N A K M K A F K K E L Q A F L 20

285 K K Ξ L Q A F L E Q V N R I G 20

289 Q A F L E Q V N R I G D G L S 20

292 L E Q V N R I G D G L S P L P 20

299 G D G L S P L P H L T E S S s 20

302 L S P L P H L T E S S S F L s 20

305 L P H L T E S s S F L s T V T 20

312 S S F L s T V T S V S R D S P 20

315 L S T V T S V s R D S P I G N 20

318 V T S V s R D s P I G N L G K 20

327 I G N L G K E L G P D L Q S R 20

389 s L Ξ M E Ξ E H L Y A L R W K 20

394 E E H L Y A L R W K E L E M H 20

404 E L E M H S L A L Q N T L H E 20

409 S L A L Q N T L H E R T W S D 20

429 Q Q E L R S L K Q N I F L F Y 20

438 N I F L F Y V K L R w L L K H 20

446 L R W L L K H W R Q G K Q M E 20

447 R W L L K H W R Q G K Q M E E 20

456 G K Q M E E Ξ G E E F T E G E 20

472 P E T L S R L G E L G V Q G G 20

519 S L R L Q T A D R G Q P H K Q 20 TABLE XXV - 185P2C9 vl HLA- DRB1*0401 (DR4D 4 ) 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

531 H K Q V V E N Q Q L F S A F K 20

537 N Q Q L F S A F K A L L E D F 20

560 R A R L R L Q Q Q Y A S D K A 20

595 E N K L G E L G S S A E S K G 20

608 K G A L K K E R E V H Q K L L 20

619 Q K L L A D S H S L V M D L R 20

645 R E K V E L L D R L D R D R Q 20

651 L D R L D R D R Q E W E R Q K 20

669 L W R I E Q L Q K E N S P R R 20

672 I E Q L Q K E N s P R R G G S 20

729 L S K L K E S D R C S A S E N 20

768 R N R L P E E E E N H K G N L 20

788 V s s M s E F Q R L M D I S P 20

819 T P P L s P D D L K Y I E E F 20

824 P D D L K Y I E E F N K S W D 20

849 G P D L W A D R T E V G R A G 20

875 S W Y L T T S V T M T T D T M 20 ■

886 T D T M T S P E H C Q K Q P L 20

901 R S H V L T E Q S G L R V L H 20

968 R D Y V E G A R R P L D S P L 20

983 C T S L G F A S P L H S L E M 20

989 A S P L H S L E M S K N L S D 20

994 S L E M S K N L S D D M K E V 20

998 S K N L S D D M K E V A F S V 20

1005 M K E V A F S V R N A I C S G 20

1040 T Q T V Q T I S V G Q T E A 20

1043 V Q T I S V G L Q T E A L R G 20

1047 S V G L Q T E A L R G S G V T 20

1057 G S G V T S S P H K C L T P K 20

1088 S R Q V A P A I E K V Q A K F 20

1153 E S P V H T T I N D G L S S L 20

1161 N D G L S s L F N I I D H S P 20

1174 S P V V Q D P F Q K G L R A G 20

1182 Q K G L R A G S R s R S A E P 20

1232 G T P V K Q D L S A P P G Y T 20

1275 P P G L H s D S H S L G D T A 20

12 G P G R D H A P S I P T S P F 18

25 P F. G D S L E S S T E L R R H 18

29 S L E S S T E L R R H L Q F V 18

30 L E S S T E L R R H L Q F V E 18

43 V E E E A E L L R R S I S E I 18

44 E E E A E L L R R S I S E I E 18

52 R S I S E I E D H N R Q L T H 18

53 S I S E I E D H N R Q T H E 18

58 E D H N R Q L T H E L S K F K 18

94 A P L Q E Ξ L K S A R L Q I S 18

103 A R L Q I S E L S G K V L K L 18

118 Q H E N H A L L S N I Q R c D 18

128 I Q R C D L A A H L G L R A P 18

156 K E S D G E E S R L P Q P K R 18

205 E L L K A R E D S E Y L V T L 18

224 Q R L E R T V E R L I T D T D 18

261 - E E Q G E G D Q Q E P Q L L G 18

268 Q Q E P Q L L G T I N A K M K 18

304 P L P H L T E S S s F L S T V 18

308 L T E S S S F L S T V T S V S 18

309 T E S S S F L S T V T S V S R 18

321 V S R D S P I G N L G K E L G 18 ^ω

a a rt H a H Ω Ω n cn rt rt a a cn KD rt α cn a > a X a O Ω CO a a n a a a 2 < a ϊ> Ω a a < KD cn Ω < a Si a a a a a a H α

TJ co n a B H a rt Ω a a a rt rt CO Ki < cn rt 3 a a Ω Ω a a Ω o r Ω ri a a < a a a KD O a a a a a a a a Ω a a CO a a CO a Ω a to a a

Ω a a a a a a a a a n a H a a Ω O a H Ω 3 a a Ω cn a iO cn a a CO a KD cn a a a a a a a a a D a a < a a 2 α 2 i a a a a H ω * s; a a a Ω a rt Ω B a a a M O O CO c rt a a ^ B a a a w a O W B a CO a a a a a « M « Ω ιo a a a o a a ιo a a a a Ω o rf* rf* a a Ω a a a B rt Ω cn H D co a a a KD B cn cn Ω a cn G s; a a a a a 2 Ω a a B s; a a M cn a a iO D H Si o a CO a 3 a a O in H

Ω a α 3 a a a a Ω H Ω a Si a a co Ω < a Tf a X a i a si B Ω 2 O a B a a a a a O a PO lO < Ω Ω a a a a a a a Ω σ en

B B cn a a a a <! a a α a < a B a a < B a a < < a a o α a O a a a B a H <ι a > a a a o a a a a > B B a D a α i

^1 a

Ω a a a a B B > a Ω a a CO a a iO a D KD a 3 a a a a a rt CO D < a H a to a a s a a X < B a a H a a a a 3 a 3 a a a o rf*

00 α c

> a a a ιo a a a K rt a cn CO rt CO a rt CO rt rt co cn cn rt a rt cn rt CO a i cn cn KD a a a a a CO G a CO rt KD co a O co a cn o a a a a cn S L vo rf* cn a cn Si rt a > H o a J TJ a rt rt a Ω a B H < a a cn < a a CO i rt a rt a a a a H O X Pi ιθ a H KD KD a J s t- o a -Q a a a a — t

O

TJ a cn a <: iO a a a <i a < a H rt J Ω KD J> CO a a a a α KD <: < Ki a a cn a a < Ω a a B O a a Ω a Ki a KD a a a a a rt a Ki a CO a H V

H in

Ω a rt a H a a co a a < a a rt a CO < a < a a 2 cn cn a ri a a ω a 2 CO a a o a a a a a a a a W O O a a a a t 1 Ω B a a ;> a a > Ω KD H o a a a a Ώ cn cn a H Ω rt 2 rt Ω co a a o a a a a a a a cn a D a Ω a a a a a α a a ω 3

Ω s a a rt a a a a α J α H Ω B a a a Ώ a H α a Ω rt a a rt rt a a B B H cn KD a to -3 3 α a Ω O a n rt < a H co a to a Ω a o ro rf* Hi

Ώ a a i Ω a a •j a Ώ a α a cn Ki > CO KD Ω o a n a K rt cn α α a a CO a a a a α co α a B a a > iO iO a a P. a s α a a ra i in

H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H cn cn cn -j 00 CO CO CO 00 CO 00 CO 00 00 00 CO CO CO 00 CO CO CO CO CO CO CO O CD CO CD CO CO CO 03 CO CO 00 CO CO CD CO CO CO CO CO CO o

_ fl⁾

3

TABLE XXV - 185P2C9 vl HLA- DRB1*0401 (DR4DW4 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

181 E E M F E K T S G F G S G K P 16 187 T S G F G S G K P S E A S E P 16 212 D S E Y L V T L K H Ξ A Q R L 16 237 T D S F L H D A G L R G G A P 16 288 L Q A F L E Q V N R I G D G L 16 385 Q T C F S L E M E E E H L Y A 16 450 L K H W R Q G K Q M E E E G E 16 499 G C G F P V G E H S P H S R V 16 538 Q Q L F S A F K A L L E D F R 16 541 F S A F K A L L E D F R A E L 16 566 Q Q Q Y A S D K A A W D V E W 16 577 D V E W A V L K C R L E Q L E 16 658 R Q E W E R Q K K E F L w R I 16 667 E F L w R I E Q L Q K E N S P 16 791 M S E F Q R L M D I S P F L P 16 834 N K S W D Y T P N R G H N G G 16 868 T E P F P D S S W Y L T T S V 16 874 S S W Y L T T s V T M T T D T 16 932 E G P F P T S R A R G S P G D 16 956 L S R W P c T S P R H S R D Y 16 985 S L G F A s P L H S L E M S K 16 1099 Q A K F E R T C C S P K Y G s 16 1108 S P K Y G S P K L Q R K P L P 16 1137 Q K G Y S E S A W A R S T T T 16 1142 E S A W A R s T T T R E S P V 16 1242 P P G Y T L T E N V A R I L N 16

18 A P S I P T s P F G D S L E S 14

33 S T E L R R H L Q F V E E E A 14

37 R R H L Q F V E E E A E L L R 14

40 L Q F V E E E A E L L R R S I 14

47 A E L L R R S I S E I E D H N 14

51 R R S I S E I E D H N R Q L T 14

65 T H E L S K F K F E P P R E P 14

93 G A P L Q E Ξ L K S A R L Q I 14 102 S A R L Q I S Ξ L S G K V L K 14 112 G K V L K L Q H E N H A L L s 14 121 N H A L L S N I Q R C D L A A 14 130 R C D L A A H L G II R A P S P 14 136 H L G L R A P S P R D S D A E 14 162 E S R L P Q P K R E G P V G G 14 171 Ξ G P V G G Ξ S D S E E M F E 14 203 P T E L L K A R Ξ D S E Y L V 14 223 A Q R L E R T V E R L I T D T 14 231 E R L I T D T D S F L H D A G 14 238 D S F L H D A G L R G G A P L 14 254 G P G L Q G E E Ξ Q G Ξ G D Q 14 295 V N R I G D G L S P L P H L T 14 324 D S P I G N L G K E L G P D L 14 339 Q S R L K E Q L E W Q L G P A 14 347 E W Q L σ P A R G D E R E S L 14 360 S L R L R A A R E L H R R A D 14 379 S H G L G G Q T C F S L E M E 14 387 C F S L E M Ξ E E H L Y A L R 14 402 W K E L E M H S L A L Q N T L 14 407 M H S L A L Q N T L H E R T W 14 424 E K N L M Q Q E L R S L K Q N 14 436 K Q N I F L F Y V K L R W L L 14 443 Y V K L R W L L K H W R Q G K 14 H tO cn cn cn ιπ ιπ ιπ ιπ ιn _π in in rf* rf* rf* TJ l0 t0 t0 _0 l i m rf* L0 vo H H oo o ~j o H ιo vo cn rf* vo tfi to ιπ to to o o co in ω

a a Ω a Ω a a a a a a. < > H cn < a a CO a o a < cn a CO rt a a B a a te tr a CO α a a KD Ω a < X a B to a a B a a H o

Ω a K KD to H •a to a co rt a rt B a Pi a a a σ a CO a α a X o < rt a a a 2 o a o a a a a Ω n S! 3 CO Ω S s; a to o < cn a Ω CO a to a

TJ a rt α a Ω B a a co rt te a te rt o o to cn a to a TJ CO a < a ri CO Ki α Ω o a ω to a a to a cn n_ KD α a a to o a a < KD a Ω a a H

LO *

3 a a a 3 < a <; H a H H < a 3 2 < H < 2 a a a H a a 2 < H < a H a a a a a a 3 < a a H a <; a < < a a < H < a a O rf* rf* a B rt n Ω Ω Ω < H a a a cn a Ω > a Ω a a a Ω o rt D rt a rt rt B rt a cn 3 o to o i CO Si a Ω a a 2 a a a o a B Ω KD tO Ω Ω O in H a a a p a W a O a o a co Ω rt Ω D ω a a 2 rt CO to CO a CO rt 2 B a CO a o o a to a a TJ 3 3 o a Si α iO a s; o α a α H Ω <: B en

KD a a a a Ω o a H Ω < a CO tO KD 2 Ω > < CO cn a > H KD X α rt a a ri a H a o a o a Ω Si a o CO KD a a Ω to KD a KD a Ω Ω KD a a 1 a rt a a a 3 o a CO H a KD cn Ω rt rt to TJ^' H a a a Ω cn < rt rt a a cn a CO a o ω to a TJ a a a H a a a < Ki a iO cn α a If*

Ω Ω CD o

< a > Ω Ω Ω B "fl a a cn a <! < a Ω Ω Ω a a Ω Ω Ω te Ω a 2 O a cn CO a a a B a a B α Ω a o a X Si B a a > > a a a KD Ω < VO rf* a a a K Ω a rt to <! X n a CO rt O Ω tO a co CO a a rt a f a rt rt rt cn a a B a a a O to CO to to Ω a X o a a a CO a •A a a to a KD - - t

O a a H rt a a Ω a <i cn a a a cn rt CO rt a Ω < CO to to Ω Ω a B cn 3 Si o a a a a ra α a o O O > a s; cn H rt KD Ω α a to a a α KD Ω H

H in rt a a a Ω a rt Ω KD a a B a CO H a a D a a α cn a a Ω < ιθ a rt α α o Ω a a a B α a CO 3 a B a M a a a a o a Ω to Ω to 1 to a a rt rt Ω a a O < a a to a cn rt Ω Ω a α a Ω a a a CO a CO Ki a : a a a a a Ω H D s a a a a a a a > B a rt a a α a LO 3 a D a a a Ω co a a < H ri a a < 2 cn a a 2 a a a Ω a Ω a a rt a rt a a a a a B CO a cn a a B a co a a B to a a > KD o Ω KD rf*

Si > a a ! cn a CO

B a a iO H Ω KD a Ω Ω a α a H a a > rt a Ω M a a CO Ω a to a a a H Ω. a a i B a a iO -S a to α rt a a to tfl

01

H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H - M r- H H H n rf* if* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* *_* rf* rf* rf* rf* rf* rf* rf* rf* it* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* rf* 0 H fl)

to

? 3 ^•?

H

TABLE XXVI - 185P2C9 vl HLA- DRB1* 1101 15 - mers

Seq. ID

POS 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

315 L S T V T S V S R D S P I G N 28 324 D S P I G N L G K E L G P D L 27 669 L W R I E Q L Q K E N S P R R 26 212 D S E Y L V T L K H E A Q R L 25 1165 S S L F N I I D H S P V V Q D 24 1178 Q D P F Q K G L R A G S R S R 24 437 Q N I F L F Y V K L R W L L K 22 648 V E L L D R L D R D R Q E W Ξ 22 791 M S E F Q R L M D I S P F L P 22 1182 Q K G L R A G S R s R s A E P 22 44 E Ξ E A E L L R R s I s E I E 21 58 E D H N R Q L T H E L s K F K 21 701 H Q G S L R M P R P V A M W P 21 719 A D S I P F E D R P L s K L K 21 776 E N H K G N L Q R A V S V S s 21 991 P L H s L E M S K N L S D D M 21 998 S K N L S D D M K E V A F S V 21 1275 P P G L H S D S H S L G D T A 21 111 S G K V L K L Q H E N H A L L 20 122 H A L L S N I Q R C D L A A H 20 162 E S R L P Q P K R E G P V G G 20 213 S E Y L V T L K H Ξ A Q R L E 20 288 L Q A F L E Q V N R I G D G L 20 289 Q A F L E Q V N R I G D G L S 20 299 G D G L S P L P H L T E S S S 20 439 I F L F Y V K L R W L L K H W 20 443 Y V K L R w L L K H W R Q G K 20 463 G E E F T E G E H P E T L S R 20 519 S L R L Q T A D R G Q P H K Q 20 645 R Ξ K V E L L D R L D R D R Q 20 693 D G N V R P F P H Q G S L R M 20 729 L S K L K E S D R C S A S E N 20 856 R T E V G R A G H E D S T E P 20 907 E Q S G L R V L H S P P A V R 20 914 L H S P P A V R R V D S I T A 20 1089 R Q V A P A I E K V Q A K F E 20 1218 I G V G S E M C R E E G G Ξ G 20 1261 E H A L K E E R R Q A A H G P 20 292 L E Q V N R I G D G L S P L P 19 311 S S S F L S T V T S V S R D s 19 395 Ξ H L Y A L R W K E L E M H s 19 499 G C G F P V G E H S P H S R V 19 577 D V E W A V L K C R L E Q L E 19 794 F Q R L M D I S P F L P E K G 19 908 Q S G L R V L H s P P A V R R 19 967 S R D Y V E G A R R P L D S P 19 985 S L G F A S P L H S L E M S K 19 1164 L S S L F N I I D H S P V V Q 19 200 E P C P T E L L K A R E D S E 18 237 T D s F L H D A G L R G G A P 18 302 L S P L P H L T E S S S F L S 18 475 L S R L G E L G V Q G G H Q A 18 548 L E D F R A E L R Ξ D E R A R 18 658 R Q E W E R Q K K E F L W R I 18 684 G G S F L C D Q K D G N V R P 18 836 S w D Y T P N R G H N G G G P 18 917 P P A V R R V D S I T A A G G 18 920 V R R V D S I T A A G G E G P 18 TABLE XXVI - 185P2C9 vl HLA- DRB1*1101 15 - mers

Seq . ID

Pos ^• 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1137 Q K G Y S E S A W A R S T T T 18

30 L Ξ S S T E L R R H L Q F V E 17

68 L S K F K F E P P R E P G W L 17

272 Q L L G T I N A K M K A F K K 17

362 R L R A A R E L H R R A D G D 17

440 F L F Y V K L R W L L K H W R 17

445 K L R W L L K H W R Q G K Q M 17

931 G E G P F P T S R A R G S P G 17

932 E G P F P T S R A R G S P G D 17

956 L S R W P C T S P R H S R D Y 17

1007 E V A F S V R N A I C S G P G 17

78 E P G W L G E G A S P G A G G 16

181 E E M F E K T S G F G S G K P 16

187 T S G F G S G K P S E A S E P 16

354 R G D E R E S L R L R A A R E 16

399 A L R W K E L E M Ή S A L Q 16

409 S L A L Q N T L H E R T W S D 16

411 A L Q N T L H E R T W S D E K 16

444 V K L R W L L K H W R Q-G K Q 16

537 N Q Q L F S A F K A L L E D F 16

541 F S A F K A L L E D F R A E L 16

552 R A Ξ L R E D Ξ R A R L R L Q 16

576 W D V E W A V L K C R E Q L 16

605 A E S K G A L K K E R E V H Q 16

612 K K Ξ R E V H Q K L L A D S H 16

689 C D Q K D G N V R P F P H Q G 16

799 D I S P F L ,P E K G L P S T s 16

800 I s P F L P E K G L P S T S S 16

830 I E E F N K S w D Y T P N R G 16

853 w A D R T E V G R A G H E D S 16

886 T D T M T S P E H C Q K Q P L 16

933 G P F P T S R A R G S P G D T 16

1005 M K E V A F S V R N A I C S G 16

1018 S G P G E L Q V K D M A C Q T 16

1028 M A C Q T N G S R T M G T Q T 16

1048 V G L Q T E A L R G S G V T S 16

1057 G S G V T S S P H K C L T P K 16

1203 G Q E T G T N S R G R S P s P 16

43 V E E E A E L L R R S I S E I 15

54 I S E I E D H N R Q L T H E L 15

62 R Q L T H E L S K F K F E P P 15

69 S K F K F E P P R E P G W L G 15

114 V L K L Q H Ξ N H A L L S N ϊ 15

161 E E S R L P Q P K R E G P V G 15

220 K H E A Q R L E R T V E R L I 15

224 Q R L E R T V E R L I T D T D 15

278 N A K M K A F K K E L Q A F L 15

305 L P H L T E S S S F L S T V T 15

363 L R A A R E L H R R A D G D T 15

372 R A D G D T G S H G L G G Q T 15

400 L R W K E L E M H S L A L Q N 15

425 K N L M Q Q E L R S L K Q N I 15

510 H S R V Q I G D H S L R L Q T 15

618 H Q K L L A D S H s L V M D L 15

674 Q L Q K E N S P R R G G S F L 15

723 P F E D R P L S K L K Ξ S D R 15

968 R D Y V E G A R R P L D S P L 15

1076 A T P V S S P S R S L R S R Q 15 TABLE XXVI - 185P2C9 vl HLA- DRB1*1101 15 - mers

Seq . ID

Pos ^■ 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

1081 s P S R S L R S R Q V A P A I 15 llll Y G S P K L Q R K P L P K A D 15

1154 s P V H T T I N D G L S S L F 15

1229 G G E G T P V K Q D L s A P P 15

1245 Y T L T E N V A R I L N K K L 15

1249 E N V A R I L N K K L L E H A 15

1254 I L N K K L L E H A L K E E R 15

1260 L E H A L K E E R R Q A A H G 15

2 E D M R G Q Q E R E G P G R D 14

7 Q Q E R E G P G R D H A P S I 14

9 E R E G P G R D H A P s I P T 14

64 L T H E L S K F K F E P P R E 14

93 , G A P L Q E E L K S A R L Q I 14

108 S E L S G K V L K L Q H E N H 14

130 R C D L A A H L G L R A P S P 14

132 D L A A H L G L R A P s P R D 14

147 S D A E S D A G K K E s D G E 14

178 S D S E E M F E K T S G F G S 14

202 C P T E L L K A R E D s E Y L 14

234 I T D T D S F L H D A G L R G 14

239 S F L H D A G L R G G A P L P 14

274 L G T I N A K M K A F K K E L 14

335 G P D L Q S R L K E Q L E W Q 14

344 E Q L E W Q L G P A R G D E R 14

364 R A A R E L H R R A D G D T G 14

388 F S L E M E E E H L Y A L R W 14

438 N I F L F Y V K L R W L L K H 14

446 L R W L L K H W R Q G K Q M E 14

469 G Ξ H P E T L S R L G E L G V 14

490 D G P D H D S D R G C G F P V 14

517 D H S L R L Q T A D R G Q P H 14

542 S A F K A L L E D F R A Ξ L R 14

544 F K A L L E D F R A E L R E D 14

625 S H S L V M D L R W Q I H H S 14

629 V M D L R W Q I H H S E K N W^' 14

630 M D L R W Q I H H S E K N W N 14

633 R W Q I H H s E K N W N R E K 14

663 R Q K K E' F L W R I E Q L Q.K 14

675 L Q K E N S P R R G G S F L C 14

699 F P H Q G S L R M P R P V A M 14

704 S L R M P R P V A M W P C A D 14

779 K G N L Q R A V s V s S M S E 14

788 V S S M S E F Q R L M D I S P 14

827 L K Y I E Ξ F N K S W D Y T P 14

837 W D Y T P N R G H N G G G P D 14

881 S V T M T T D T M T s P E H C 14

923 V D S I T A A G G E G P F P T 14

940 A R G S P G D T K G G P P E P 14

950 G P P E P M L s R W P c T S P 14

980 S P L C T S L G F A S P L H S 14

1063 S P H K c L T P K A G G G A T 14

1065 H K C L T P K A G G G A T P V 14

1102 F E R T c C S P K Y G S P K L 14

1110 K Y G S P K L Q R K P L P K A 14

1115 K L Q R K P L P K A D Q P N N 14

1122 P K A D Q P N N R T S P G M A 14

1130 R T S^'P G M A Q K G Y S E S A 14

1144 A w A R S T T T R E S P V H T 14 TABLE XXVI - 185P2C9 vl HLA- DRB1*1101 15 - mers

Seq. ID

Pos - 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1205 E T G T N S R G R S P S P I G 14 1257 K K L L E H A L K E E R R Q A 14 1265 K E E R R Q A A H G P P G L H 14 1271 A A H G P P G L H S D S H S L 14 1 M E D M R G Q Q E R E G P G R 13

97 Q E E L K S A R L Q I S E L S 13

104 R L Q I S E L S G K V L K L Q 13

127 N I Q R C D L A A H L G L R A 13

131 C D L A A H L G L R A P S P R 13

136 H L G L R A P S P R D S D A E 13

180 S E E M F E K T S G F G S G K 13

238 D S F L H D A G L R G G A P L 13

295 V N R I G D G L S P L P H L T 13

340 S R L K E Q L E W Q L G P A R 13

355 G D E R E S L R L R A A R E L 13

429 Q Q E L R S L K Q N I F L F Y 13

478 L G E L G V Q G G H Q A D G P 13

498 R G C G F P V G Ξ H S P H S R 13

598 L G E L G S S A E S K G A L K 13

626 H S L V M D L R W Q I H H S E 13

797 L M D I S P F L P E K G L P S 13

812 T S S K E D V T P P L S P D D 13

849 G P D L W A D R T E V G R A G 13

899 P L R S H V L T E Q S G L R V 13

949 G G P P E P M L S R W P C T S 13

976 R P L D S P L C T S L G F A S 13

989 A S P L H S L E M S K N L S D 13

1002 S D D M K E V A F S V R N A I 13

1040 T Q T V Q T I S V G L Q T E A 13

1043 V Q T I S V G L Q T E.A L R G 13

1047 S V G L Q T E A L R G S G V T 13

1113 S P K L Q R K P L P K A D Q P 13

1161 N D G L S S L F N I I D H S P 13

1173 H S P V V Q D P F Q K G L R A 13

1241 A P P G Y T L T E N V A R I L 13

1248 T E N V A R I L N K K L L E H 13

TABLE XXIII - 185P2C9 v2 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

104 R L Q I S E L S G K V L K L Q 33 908 Q S G L R V L H S P P A V R R 32 478 L G E L G V Q G G H Q A D G P 31 696 V R P F P H Q G S L R M P R P 31 902 S H V L T E Q S G L R V L H S 31 181 E E M F E K T S G F G S G K P 30 917 P P A V R R V D S I T A A G G 30 932 E G P F P T s R A R G S P G D 29 39 H L Q F V E E E A E L L R R S 28 358 R E S L R L R A A R Ξ L H R R 28 399 A L R W K E L E M H S L A L Q 28 794 F Q R L M D I S P F L P Ξ K G 28 923 V D S I T A A G G E G P F P T 28 68 L S K F K F E P P R E P G W L 27 472 P E T L S R_. L G E L G V Q G G 27 1065 H K c L T P K A G G G A T P V 27 246 L R G G A P L P G P G L Q G E 26 TABLE XXIII - 185P2C9 v2 HLA- DRB1*0101 15 - mers

Seq. ID

Pos • 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

327 I G N L G K E L G P D L Q S R 26

429 Q Q E L R S L K Q N I F L F Y 26

711 V A M W P C A D A D S I P F E 26

791 M S E F Q R L M D I s P F L P 26

800 I S P F L P E K G L P s T S S 26

1007 E V A F S V R N A I c s G P G 26

1049 G L Q T E A L R G S G V T S S 26

136 H L G L R A P S P R D s D A E 25

394 E E H L Y A L R W K E L E M H 25

517 D H S L R L Q T A D R G Q P H 25

560 R A R L R L Q Q Q Y A S D K A 25

702 Q G S L R M P R P V A M W P C 25

873 D S S W Y L T T S V T M T T D 25

956 L S R W P C T S P R H S R D Y 25

1080 s s- P S R S L R s R Q V A P A 25

1092 A P A I E K V Q A K F E R T C^' 25

79 P G W L G E G A S P G A G G G 24

112 G K V L K L Q H E N H A L L S 24

130 R C D L A A H L G L R A P S P 24

133 L A A H L G L R A P S P R D S 24

187 T S G F G S G K P S E A S E P 24

240 F L H D A G L R G G A P L P G 24

268 Q Q E P Q L L G T I N A K M K 24

292 L Q Q V N R I G D G L S P L P 24

302 L S P L P H L T Ξ S S S F L S 24

311 S S s F L S T V T S V S R D S 24

424 Ξ K N L M Q Q E L R S L K Q N 24

508 S P H S R V Q I G D H S L R L 24

658 R Q E W E R Q K K E F L W R I 24

669 L W R I E Q L Q K E N S P R R 24

704 S L R M P R P V A M W P c A D 24

740 A S E N L Y L D A L S L D D E 24

777 N H K G N L Q R A V S V S S M 24

803 F L P E K G L P S T S S K E D 24

920 V R R V D S I T A A G G E G P 24

979 D S P L C T s L G F A S P L H 24

994 S L E M S K N L S D D M K E V 24

100 L K S A R L Q I S Ξ L S G K V 23

271 P Q L L G T I N A K M K A F K 23

305 L P H L T E s S S F L S T V T 23

309 T E S S S F L S T V T S V S R 23

315 L S T V T S V S R D S P I G N 23

339 Q S R L K E Q L E W Q L G P A 23

438 N I F L F Y V K L R W L L K H 23

595 E N K L G E L G S S A E S K G 23

983 C T s L G F A S P L H S L E M 23

1032 T^'N G S R T M G T Q T V Q T I 23

1044 Q T I S V G L Q T E A L R G S 23

94 A P L Q E E L K S A R L Q I S 22

122 H A L L S N I Q R C D L A A H 22

227 E R T V E R L I T D T D S F L 22

295 V N R I G D G L s P L P H L T 22

343 K E Q L E W Q L G P A Q G D E 22

379 S H G L G G Q T C F S L E M E 22

532 K Q V V E N Q Q L F S A F K A 22

562 R L R L Q Q Q Y A S D K A A W 22

592 E K T E N K L G E L G S S A.E 22

619 Q K L L A D S H S L V M D L R 22 TABLE XXIII - 185P2C9 v2 HLA- DRB1*0101 15 - mers

Seq. ID

Pos • 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

685 G S F L C D Q K D G N V R P F 22

779 K G N L Q R A V S V S S M S E 22

816 E D V T P P L S P D D L K Y I 22

894 H C Q K Q P L R S H V L T E Q 22

952 P E P M L S R W P C T S P R H 22

986 L G F A S P L H S L E M S K N 22

1035 S R T M G T Q T V Q T I S V G 22

1084 R S L R S R Q V A P A I E K V 22

57 I E D H N R Q L T H E L S K F 21

263 Q G E G D Q Q E P Q L L G T I 21

281 M K A F K K E L Q A F L Q Q V 21

293 Q Q V N R I G D G L S P L P H 21

345 Q L E W Q L G P A Q G D Ξ R E 21

407 M H S L A L Q N T L H E R T W 21

440 F L F Y V K L R W L L K H W R 21

830 I Ξ E F N K S W D Y T P N R G 21

878 L T T S V T M T T D T M T S P 21

949 G G P P E P M L S R w P C T S 21

15 R D H A P s I P T S P F G D S 20

312 S S F L S T V T S V s R D S P 20

401 R W K Ξ L E M H S L A L Q N T 20

631 D L R W Q I H H S E K N W N R 20

693 D G N V R P F P H Q G S L R M 20

798 M D I S P F L P E K G L P S T 20

812 T S S K E D V T P P L S P D D 20

839 Y T P N R G H N G G G P D L W 20

898 Q P L R S H V L T E Q S G L R 20

967 S R D Y V E G A R R P L D S P 20

1009 A F S V R N A I C S G P G E L 20

1037 T M G T Q T V Q T I S V G L Q 20

1052 T E A L R G S G V T S S P H K 20

78 E P G W L G E G A S P G A G G 19

215 Y L V T L K H E A Q R L E R T 19

235 T D T D S F L H D A G L R G G 19

291 F L Q Q V N R I G. D G L S P L 19

475 L S R L G E L G V Q G G H Q A 19

541 F S A F K A L L E D F R A Ξ L 19

552 R A E L R E D E R A R L R L Q 19

577 D V E W A V L K C R L E Q L E 19

640 E K N W N R E K V E L L D R L 19

721 S I P F E D R P L S K L K Ξ S 19

836 S w D Y T P N R G H N G G G P 19

1047 S V G L Q T E A L R G S G V T 19

1110 K Y G S P K L Q R K P L P K A 19

58 E D H N R Q L T H E L S K F K 18

65 T H E L S K F K F E P P R E P 18

75 P P R E P G W L G E G A S P G 18

77 R E P G w L G E G A S P G A G 18

83 G E G A S P G A G G G A P L Q 18

111 S G K V L K L Q H E N H A L L 18

123 A L L S N I Q R C D L A A H L 18

126 S N I Q R c D L A A H L G L R 18

131 C D L A A H L G L R A P S P R 18

162 Ξ S R L P Q P K R E G P V G G 18

185^' E K T S G F G S G K P S E A S 18

190 F G S G K P S E A S Ξ P C P T 18

201 P C P T E L L K A R E D S E Y 18

212 D S E Y L V T L K H E A Q R L. 18 TABLE XXIII - 185P2C9 v2 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

228 R T V E R L I T D T D S F L H 18 237 T D S F L H D A G L R G G A P 18 241 L H D A G L R G G A P L P G P 18 435 K Q N I F L F Y V K L R W L 18 439 I F L F Y V K L R W L L K H W 18 443 Y V K L R W L L K H W R Q G K 18 461 E E G E E F T E G E H P E T L 18 481 L G V Q G G H Q A D G P D H D 18 491 G P D H D S D R G C G F P V G 18 535 V E N Q Q L F S A F K A L L Ξ 18 538 Q Q F S A F K A L L E D F R 18 542 S A F K A L L E D F R A E L R 18 566 Q Q Q Y A S D K A A W D V E W 18 573 K A A W D V Ξ W A V L K C R L 18 579 E W A V L K C R L E Q L E E K 18 584 K C R L E Q L E E K T E N K L 18 599 G E L G S S A E S K G A L K K 18 615 R E V H Q K L L A D S H S L V 18 616 E V H Q K L L A D S H S L V M 18 625 S H S L V M D L R W Q I H H S 18 726 D R P L s K L K E S D R C S A 18 764 E R E F R N R L P Ξ E E E N H 18 881 S V T M T T D T M T S P E H C 18 943 S P G D T K G G P P E P M L S 18 966 H S R D Y V Ξ G A R R P L D S 18 980 S P L C T S L G F A S P L H S 18 989 A S P L H S L E M S K N L S D 18 1001 L S D D M K E V A F S V R N A 18 1057 G S G V T s S P H K C L T P K 18 1067 C L T P K A G G G A T P V S S 18 1068 L T P K A G G G A T P V S S P 18 1083 S R S L R S R Q V A P A I E K 18 1105 T C C S P K Y G S P K L Q R K 18

2 E D T R G Q Q E R E G P G R D 17

19 P S I P T S P F G D S L Ξ S S 17

43 V E E E A E L L R R S I S E I 17

44 E E E A E L L R R S I s Ξ I E 17

62 R Q L T H E L S K F K F E P P 17

76 P R E P G W L G E G A S P G A 17

84 E G A S P G A G G G A P L Q E 17

85 G A S P G A G G G A P L Q E Ξ 17

91 G G G A P L Q E E L K S A R L 17

97 Q E E L K S A R L Q I S E L S 17 152 D A G K K E S D G E E S R L P 17 179 D S Ξ E M F E K T S G F G S G 17 213 S E Y L V T L K H E A Q R L E 17 223 A Q R L E R T V E R L I T D T 17 229 T V E R L I T D T D S F L H D 17 242 H D A G L R G G A P L P G P G 17 267 D Q Q E P Q L L G T I N A K M 17 270 E P Q L L G T I N A K M K A F 17 275 G T I N A K M K A F K K E L Q 17 282 K A F K K E L Q A F L Q Q V N 17 288 L Q A F L Q Q V N R I G D G L 17 323 R D S P I G N L G K E L G P D 17 324 D S P I G N L G K E L G P D L 17 355 G D E R E S L R L R A A R E L 17 371 R R A D G D T G S H G L G G Q 17 TABLE XXIII - 185P2C9 v2 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

376 D T G S H G L G G Q T C F S L 17

421 W S D E K N L M Q Q E L R S L 17

448 W L L K H W R Q G K Q M E Ξ E 17

455 Q G K Q M Ξ E E G E E F T E G 17

497 D R G C G F P V G E H S P H S 17

523 Q T A D R G Q P H K Q V V E N 17

531 H K Q V V E N Q Q L F S A F K 17

587 L E Q L E Ξ K T E N K L G E L 17

611 L K K Ξ R E V H Q K L L A D S 17

645 R E K V E L L D R L D R D R Q 17 ^•

677 K E N S P R R G G S F L C D Q 17

708 P R P V A M W P C A D A D S I 17

719 A D S I P F E D R P L S K L K 17

729 L S K L K Ξ S D R C S A S E N 17

792 S E F Q R L M D I S P F L P E 17

819 T P P L S P D D L K Y I E E F 17

841 P N R G H N G G G P D L W A D 17

856 R T E V G R A G H E D S T E P 17

925 S I T A A G G E G P F P T S R 17

953 E P M L S R W P C T S P R H S 17

975 R R P L D S P L C T S L G F A 17

977 P L D S P L C T S L G F A S P 17

981 P L C T S L G F A S P L H S L 17

1005 M K E V A F S V R N A I C S G 17

1017 C S G P G E L Q V K D M A C Q 17

1039 G T Q T V Q T I S V G L Q T E 17

1043 V Q T I S V G L Q T E A L R G 17

1061 T S S P H K C L T P K A G G G 17

1113 S P K L Q R K P L P K A D Q P 17

1116 L Q R K P L P K A D Q P N N R 17

10 R E G P G R D H A P s I P T S 16

11 E G P G R D H A P S I P T s P 16

18 A P S I P T S P F G D s L Ξ S 16

25 P F G D S L E S S T E L R R H 16

37 R R H L Q F V E E Ξ A Ξ L L R 16

46 Ξ A E L L R R S I S E I E D H 16

47 A E L L R R S I s E I E D H N 16

61 N R Q L T « E L s K F K F E P 16

99 E L K S A R L Q I S E L S G K 16

114 V L K L Q H E N H A L L S N I 16

115 L K L Q H Ξ N H A L L S N I Q 16

127 N I Q R C D L A A H L G L R A 16

155 K K E S D G E E S R L P Q P K 16

167 Q P K R E G P V G G E s D S E 16

184 F E K T S G F G S G K P S E A 16

230 V E R L I T D T D S F L H D A 16

243 D A G L R G G A P L P G P G L 16

251 P L P G P G L Q G E E E Q G E 16

289 Q A F L Q Q V N R I G D G L S 16

299 G D G L S P L P H L T Ξ S S S 16

303 S P L P H L T E S S S F L S T 16

314 F L S T V T S V S R D S P I G 16

318 V T S V S R D S P I G N L G K 16

328 G N L G K E L G P D L Q S R L 16

331 G K Ξ L G P D L Q S R L K E Q 16

335 G P D L Q S R L K E Q L E W Q 16

347 E W Q Ii G P A Q G D E R Ξ S L 16

357 E R E S L R L R A A R E L H R 16 TABLE XXIII - 185P2C9 v2 HLA- DRB1*0101 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

389 S L E M E E E H L Y A L R W K 16 391 E M E E E H L Y A L R W K E L 16 404 E L E M H S L A L Q N T L H E 16 431 E L R S L K Q N I F L F Y V K 16 493 D H D S D R G C G F P V G E H 16 509 P H S R V Q I G D H S L R L Q 16 514 Q I G D H S L R L Q T A D R G 16 519 S L R L Q T A D R G Q P H K Q 16 537 N Q Q L F s A F K A L L E D F 16 544 F K A L L E D F R A E L R E D 16 545 K A L L E D F R A Ξ L R E D Ξ 16 559 E R A R L R L Q Q Q Y A S D K 16 567 Q Q Y A S D K A A w D V E W A 16 594 T E N K L G E L G s S A E S K 16 614 E R E V H Q K L L A D S H S L 16 618 H Q K L L A D S H S L V M D L 16 626 H S L V M D L R W Q I H H S E 16 629 V M D L R W Q I H H s E K N W 16 648 V E L L D R L D R D R Q E W E 16 667 E F L W R I Ξ Q L Q K E N S P 16 672 I E Q L Q K E N S P R R G G S 16 676 Q K E N S P R R G G S F L C D 16 699 F P H Q G s L R M P R P V A M 16 700 P H Q G S L R M P R P V A M W 16 710 P V A M W P C A D A D S I P F 16 733 K E S D R C S A S E N L Y L D 16 738 C S A S E N L Y L D A L S L D 16 742 Ξ N L Y L D A L S L D D E P E 16 743 N L Y L D A L S L D D Ξ P E E 16 748 A L S L D D E P E E P P A H R 16 752 D D E P E E P P A H R P E R E 16 776 E N H K G N L Q R A V S V S S 16 780 G N L Q R A V S V S S M S E F 16 783 Q R A V S V S S M S E F Q R L 16 802 P F L P Ξ K G L P S T S S K E 16 824 P D D L K Y I E E F N K S W D 16 846 G G G P D L W A D R T E V G 16 905 L T E Q S G L R V L H s P P A 16 909 S G L R V L H S P P A V R R V 16 911 L R V L H S P P A V R R V D S 16 941 R G S P G D T K G G P P E P M 16 972 E G A R R P L D S P L C T S L 16 990 S P L H S L E M S K N L S D D 16 998 S K N L S D D M K E V A F S V 16 1013 R N A I C S G P G E L Q V K D 16 1024 Q V K D M A C Q T N G S R T M 16 1027 D M A C Q T N G s R T M G T Q 16 1051 Q T E A L R G S G V T S S P H 16 1062 S S P H K C L T P K A G G G A 16 1075 G T P V S S P S R S L R S R 16 1097 K V Q A K F E R T C C S P K Y 16 llll Y G S P K L Q R K P L P K A D 16

TABLE XXIV - 185P2C9 v2 HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num. 544 F K A L L E D F R A E L R E D 38 TABLE XXIV - 185P2C9 v2 HLA- DRB1*0301 (DR17) 15 - mers

Seq . ID

Pos • 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

552 R A E L R E D E R A R L R L Q 38

331 G K E L G P D L Q S R L K E Q 36

230 V Ξ R L I T D T D S F L H D A 29

625 S H S L V M D L R W Q I H H S 29

429 Q Q E L R S L K Q N I F L F Y 28

339 Q S R L K E Q L Ξ W Q L G P A 27

387 C F S L E M E E E H L Y A L R 27

537 N Q Q L F S A F K A L L E D F 27

587 L E Q L Ξ E K T E N K L G E L 27

746 L D A L S L D D Ξ P E E P P A 27

53 S I S E I Ξ D H N R Q L T H E 26

173 P V G G E S D S E Ξ M F E K T 26

335 G P D L Q S R L K E Q L E W Q 26

651 L D R L D R D R Q Ξ W Ξ R Q K 26

684 G G- S F L C D Q K D G N V R P 26

998 S K N L S D D M K E V A F S V 26

281 M K A F K K Ξ L Q A F L Q Q V 25

350 L G P A Q G D Ξ R E S L R L R 25

122 H A L S N I Q R c D L A A H 24

439 I F L F Y V K L R W L L K H W 24

618 H Q K L L A D S H s L V M D L 24

741 S E N L Y L D A L s L D D E P 22

126 S N I Q R C D L A A H L G L R 21

237 T D S F L H D A G L R G G A P 21

318 V T S V S R D S P I G N L G K 21

793 E F Q R L M D I S P F L P E K 21

819 T P P L S P D D L K Y I E E F 21

881 S V T M T T D T M T S P E H C 21

1057 G S G V T S S P H K c L T P K 21

39 H L Q F V E E E A Ξ L L R R S 20

104 R L Q I s E L S G K V L K L Q 20

114 V L K L Q H E N H A L L S N I 20

204 T E L L K A R E D S E Y L V T 20

206 L L K A R E D S E Y L V T L K 20

270 E P Q L L G T I N A K M K A F 20

294 Q V N R I G D G L S P L P H L 20

358 R E S L R L R A A R E L H R R 20

389 S L E M E E E H L Y A L R W K 20

417 H E R T W S D E K N L M Q Q E 20

432 L R S L K Q N I F L F Y V K L 20

511 S R V Q I G D H S L R L Q T A 20

579 E W A V L K C R L E Q L E E K 20

580 A V L K C R L E fl L E E K T 20

811 S T S S K E D V T P P L S P D 20

849 G P D L w A D R T E V G R A G 20

860 G R A G H E D S T E P F P D S 20

867 S^'T E P F P D S s W Y L T T S 20

975 R R P L D S P L c T S L G F A 20

40 L Q F V E E E A E L L R R S I 19

93 G A P L Q E E L K S A R L Q I 19

112 G K V L K L Q H E N H A L L S 19

121 H A L L S N I Q R C D L A A 19

214 E Y L V T L K H E A Q R L E R 19 ^'

278 N A K M K A F K K E L Q A F L 19

315 L S T V T S V S R D S P I G N 19

327 I G N L G K E L G P D L Q S R 19

424 E K N L M Q Q E L R S L K Q.N 19

438 N I F L F Y V K L R W L L K H 19 TABLE XXIV - 185P2C9 v2 HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

446 L R W L L K H W R Q G K Q M E 19

491 G P D H D S D R G C G F P V G 19

531 H K Q V V E N Q Q L F S A F K 19

560 R A R L R L Q Q Q Y A S D K A 19

768 R N R L P E E E E N H K G N L 19

783 Q R A V S V S S M S E F Q R L 19

824 P D D L K Y I E E F N K S W D 19

992 L H S L E M s K N L S D D M K 19

1045 T I S V σ L Q T E A L R G S G 19

1076 A T P V S S P s R S L R S R Q 19

27 G D S L E S s T E L R R H L Q 18

33 S T E L R R H L Q F V E E E A 18

37 R R H L Q F V E E E A E L L R 18

54 I S E I E D H N R Q L T H E L 18

113 K V L K L Q H E N H A L L S N 18

153 A G K K E S D G E E S R L P Q 18

203 P T E L L K A R E D S E Y L V 18

216 L V T L K H E A Q R L E R T V 18

254 G P G L Q G E E E Q G E G D Q 18

285 K K E L Q A F L Q Q V N R I G 18

324 D s P I G N L G K E L G P D L 18

360 S L R L R A A R E L H R R A D 18

379 S H G L G G Q T C F S L E M E 18

386 T C F S L E M E Ξ E H L Y A L 18

394 Ξ E H L Y A L R W K E L E M H 18

425 K N L M Q Q E L R S L K Q N I 18

443 Y V K L R w L L K H W R Q G K 18

447 R W L L K H W R Q G K Q M E E 18

456 G K Q M E E E G E E F T E G E 18

472 P E T L S R L G E L G V Q G G 18

558 D E R A R L R L Q Q Q Y A S D 18

598 L G E L G S S A E S K G A L K 18

608 K G A L K K E R E V H Q K L L 18

633 R W Q I H H S E K N W N R Ξ K 18

645 R E K V Ξ L L D R L D R D R Q 18

665 K K E F L W R I E Q L Q K E N 18

666 K E F L W R I E Q L Q K E N S 18

687 F L C D Q K D G N V R P F P H 18

721 S I P F E D R P L S K L K E S 18

729 L s K L K E S D R C S A S E N 18

734 E s D R C S A S E N L Y L D A 18

785 A V S V s S M S Ξ F Q R L M D 18

788 V s S M s E F Q R L M D I S P 18

1005 M K E V A F S V R N A I c S G 18

1091 V A P A I E K V Q A K F E R T 18

TABLE XXV - 185P2C9 v2 HLA- DRB1*0401 (DR4D 4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

281 M K A F K K E L Q A F L Q Q V 28 418 E R T W S D E K N L M Q Q E L 28 61 N R Q L T H E L S K F K F E P 26 114 V L K L Q H E N H A L L S N I 26 122 H A L S N I Q R C D L A A H 26 213 S E Y L V T L K H E A Q R L E 26 285 K K E L Q A F L Q Q V N R I G 26 331 G K E L G P D L Q S R L K E Q 26 TABLE XXV - 185P2C9 v2 HLA- DRB1*0401 (DR4DW4 ) 15 - mers

Seq. ID

Pos • • 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

425 K N L M Q Q E L R S L K Q N I 26

544 F K A L L E D F R A E L R E D 26

552 R A E L R E D E R A R L R L Q 26

618 H Q K L L A D S H S L V M D L 26

625 S H S L V M- D L R W Q I H H S 26

648 V Ξ L L D R L D R D R Q E W E 26

666 K E F L W R I E Q L Q K E N S 26

693 D G N V R P F P H Q G S L R M 26

719 A D S I P F E D R P L S K L K 26

779 K G N L Q R A V S V S s M S E 26

856 R T E V G R A G H Ξ D s T E P 26

908 Q S G L R V L H S P P A V R R 26

917 P P A V R R V D S I T A A G G 26

1118 R K P L P K A D Q P N N R P G 26

39 H L Q F V E E E A E L L R R S 22

311 S S S F L s T V T S V S R D S 22

399 A L R W K E L E M H s L A L Q 22

440 F L F Y V K L R W L L K H W R 22

463 G Ξ E F T E G E H P Ξ T L S R 22

548 L E D F R A E L R Ξ D E R A R 22

573 K A A W D V E W A V L K C R L 22

631 D L R W Q I H H S E K N W N R 22

640 E K N W N R E K V E L L D R L 22

684 G G S F L C D Q K D G N V R P 22

696 V R P F P H Q G S L R M P R P 22

711 V A M W P C A D A D S I P F E 22

742 E N L Y L D A L S L D D E P E 22

800 I S P F L P E K G L P S T S S 22

826 D L K Y I E E F N K S W D Y T 22

873 D S S W Y L T T S V T M T T D 22

967 S R D Y V E G A R R P L D S P 22

54 I S E I E D H N R Q L T H E L 20

79 P G W L G E G A S P G A G G G 20

97 Q E E L K S A R L Q I S Ξ L S 20

104 R L Q I S E L S G K V L K L Q 20

111 S G K V L K L Q H E N H A L L 20

216 L V T L K H E A Q R L E R T V 20

227 E R T V E R L I T D T D S F L 20

230 V E R L I T D T D S F L H D A 20

270 E P Q L L G T I N A K M K A F 20

271 P Q L L G T I N A K M K A F K 20

278 N A K M K A F K K E L Q A F L 20

289 Q A F L Q Q V N R I G D G L S^' 20

292 Q Q V N R I G D G L S P L P 20

299 G D G L S P L P H L T E S S S 20

302 L S P L P H L T E S S S F L S 20

305 L P H L T E S S S F L S T V T 20

312 S S F L s T V T S V S R D S P 20

315 L S T V T S V S R D S P I G N 20

318 V T S V s R D S P I G N L G K 20

327 I G N L G K E L G P D L Q S R 20

389 S L E M E E E H L Y A L R W K 20

394. E E H L Y A L R W K E L E M H 20

404 E L E M H S L A L Q N T L H E 20

409 S L A L Q N T L H E R T W S D 20

429 Q Q E L R S L K Q N I F L F Y 20

438 N I F L F Y V K L R w L L K H 20

446 L R W L L K H W R Q G K Q M E 20 TABLE XXV - 185P2C9 v2 HLA- DRB1*0401 (DR4D 4 ) 15 - mers

Pos 8 Seq. ID

1 2 3 4 5 6 7 9 0 1 2 3 4 5 score Num.

447 R W L L K H W R Q G K Q M E E 20

456 G K Q M E E E G E E F T E G E 20

472 P E T L S R L G E L G V Q G G 20

519 S L R L Q T A D R G Q P H K Q 20

531 H K Q V V E N Q Q L F s A F K 20

537 N Q Q L F S A F K A L L E D F 20

560 R A R L R L Q Q Q Y A s D K A 20

595 E N K L G E L G S s A E S K G 20

608 K G A L K K E R E V H Q K L L 20

619 Q K L L A D S H S L V M D L R 20

645 R E K V Ξ L L D R L D R D R Q 20

651 L D R L D R D R Q E W E R Q K 20

669 L W R I E Q L Q K Ξ N S P R R 20

672 I E Q L Q K E N S P R R G G S 20

729 L S K L K E S D R C S A S E N 20

768 R N R L P E E E E N H K G N L 20

788 V S S M s E F Q R L M D I S P 20

819 T P P L s P D D L K Y I E E F 20

824 P D D L K Y I E E F N K S W D 20

849 G P D L w A D R T E V G R A G 20

875 S W Y L T T S V T M T T D T M 20

886 T D T M T S P E H C Q K Q P L 20

901 R S H V L T E Q S G L R V L H 20

968 R D Y V Ξ G A R R P L D S P L 20

983 C T S L G F A S P L H S L E M 20

989 A S P L H S L E M S K N L S D 20

994 S L E M S K N L S D D M K E V 20

998 S K N L S D D M K E V A F S V 20

1005 M K E V A F S V R N A I C S G 20

1040 T Q T V Q T I S V G L Q T E A 20

1043 V Q T I S V G L Q T E A L R G 20

1047 S V G L Q T E A L R G S G V T 20

1057 G S G V T S S P H K C L T P K 20

1088 S R Q V A P A I E K V Q A K F 20

12 G P G R D H A P S I P T S P F 18

25 P F G D S L E s S T E L R R H 18

29 S L E S S T Ξ L R R H L Q F V 18

30 L E S S T E L R R H L Q F V E 18

43 V E E E A E L L R R S I S E I 18

44 Ξ Ξ E A Ξ L L R R S I S E I Ξ 18

52 R S I S E I E D H N R Q L T H 18

53 S I s E I E D H N R Q L T H E 18

58 E D H N R Q L T H E L S K F K 18

94 A P L Q E E L K S A R L Q I S 18

103 A R L Q I S E L S G K V L K L 18

118 Q H E N H A L L S N I Q R c D 18

128 I Q R C D L A A H L G L R A P 18

156 K E S D G E E S R L P Q P K R 18

205 E L L K A R E D S E Y L V T L 18

224 Q R L E R T V E R L I T D T D 18

261 E E Q G E G D Q Q E P Q L L G 18

268 Q Q E P Q L L G T I N A K M K 18

304 P L P H L T Ξ S s S F L S T V 18

308 L T E S S S F L s T V T S V s 18

309 T Ξ S S S F L S T V T S V S R 18

321 V S R D S P I G N L G K E L G 18

332 K E L G P D L Q s R L K E Q L 18

350 L G P A Q G D Ξ R Ξ S L R L R 18 TABLE XXV - 185P2C9 v2 HLA- DRB1*0401 (DR4D 4 ) 15 - mers

Pos 1 2 3 4 5 6 7 8 9 :

0 1 2 3 4 Seq.

5 score Num.

362 R L R A A R E L H R R A D G D 18

388 F S L E M E E E H L Y A L R W 18

400 L R W K E L E M H S L A L Q N 18

405 L E M H S L A L Q N T L H E R 18

421 W S D E K N L M Q Q E L R S L 18

426 N L M Q Q E L R S L K Q N I F 18

444 V K L R W L L K H W R Q G K Q 18

500 C G F P V G E H S P H S R V Q 18

511 S R V Q I G D H S L R L Q T A 18

529 Q P H K Q V V E N Q Q L F s A 18

534 V V Ξ N Q Q L F S A F K A L L 18

559 E R A R L R L Q Q Q Y A S D K 18

586 R L E Q L E E K T E N K L G E 18

599 G E L G S S A Ξ S K G A L K K 18

610 A L K K E R E V H Q K L L A D 18

611 L K K E R E V H Q K L L A D S 18

617 V H Q K L L A D S H S L V M D 18

630 M D L R W Q I H H S E K N W N 18

663 R Q K K E F L W R I Ξ Q L Q K 18

760 A H R P E R E F R N R L P E Ξ 18

770 R L P E Ξ Ξ E N H K G N L Q R 18

776 E N H K G N L Q R A V S V S S 18

787 S V S S M S E F Q R L M D I S 18

793 E F Q R L M D I S P F L P E K 18

803 F L P E K G L P S T S S K E D 18

811 S T S S K E D V T P P L S P D 18

848 G G P D L W A D R T E V G R A 18

867 S T E P F P D S S W Y L T T S 18

872 P D S S W Y L T T S V T M T T 18

894 H C Q K Q P L R s H V L T E Q 18

898 Q P L R s H V L T E Q S G L R 18

914 L H S P P A V R R V D s I T A 18

929 A G G E G P F P T S R A R G S 18

986 L G F A S P L H S L E M S K N 18

990 S P L H S L E M S K N L S D D 18

1003 D D M K E V A F S V R N A I C 18

1037 T M G T Q T V Q T I S V G L Q 18

1044 Q T I S V G L Q T E A L R G S 18

1080 S S P S R S L R S R Q V A P A 18

1097 K V Q A K F E R T C C S P K Y 18

1110 K Y G S P K L Q R K P L P K A 18

439 I F L F Y V K L R W L L K H W 17

23 T S P F G D S L E S S T E L R 16

68 L s K F K F E P P R E P G W L 16

78 E P G W L G Ξ G A S P G A G G 16

181 E E M F E K T S G F G S G K P 16

187 T s G F G S G K P S Ξ A S E P 16

212 D s E Y L V T L K H E A Q R L 16

237 T D S F L H D A G L R G G A P 16

288 L Q A F L Q Q V N R I G D G L 16

385 Q T c F S L E M E E E H L Y A 16

450 L K H W R Q G K Q M E E E G E 16

499 G c G F P V G E H S P H S R V 16

538 Q Q L F S A F K A L L Ξ D F R 16

541 F S A F K A L L Ξ D F R A Ξ L 16

566 Q Q Q Y A S D K A A W D V E W 16

577 D V E W A V L K C R L E Q L E 16

658 R Q E W E R Q K K E F L W R I 16 TABLE XXV - 185P2C9 V2 HLA- DRB1*0401 (DR4DW4 ) 15 - mers

Seq . ID

Pos - 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

667 E F L W R I E Q L Q K E N S P 16

791 M S E F Q R L M D I S P F L P 16

834 N K S W D Y T P N R G H N G G 16

868 T E P F P D S S W Y L T T S V 16

874 s s W Y L T T S V T M T T D T 16

932 E G P F P T s R A R G S P G D 16

956 L S R W P c T S P R H S R D Y 16

985 S L G F A s P L H S L Ξ M S K 16

1099 Q A K F E R T c C S P K Y G S 16

1108 S P K Y G S P K L Q R K P L P 16

18 A P s I P T s P F G D s L E S 14

33 S T E L R R H L Q F V E E E A 14

37 R R H L Q F V Ξ E E A E L L R 14

40 L Q F V E E E A E L L R R S I 14

47 A Ξ- L L R R s I S E I E D H N 14

51 R R S I S Ξ I Ξ D H N R Q L T^' 14

65 T H E L S K F K F E P P R E P 14

93 G A P L Q E E L K S A R L Q I 14

102 S A R L Q I S E L S G K V L K 14

112 G K V L K L Q H E N H A L L S 14

121 N H A L L S N I Q R C D L A A 14

130 R C D L A A H L G L R A P S P 14

136 H L G L R A P S P R D S D A E 14

162 E S R L P Q P K R E G P V G G 14

171 E G P V G G E S D S E E M F E 14

203 P T E L L K A R E D S E Y L V 14

223 A Q R L Ξ R T V E R L I T D T 14

231 E R L I T D T D S F L H D A G 14

238 D S F L H D A G L R G G A P L 14

254 G P G L Q G Ξ Ξ E Q G E G D Q 14

295 V N R I G D G L S P L P H L T 14

324 D S P I G N L G K E L G P D L 14

339 Q S R L K Ξ Q L E W Q L G P A 14

343 K E Q L E W Q L G P A Q G D E 14

347 E W Q L G P A Q G D E R E S L 14

360 S L R L R A A R E L H R R A D 14

379 S H G L G G Q T C F S L E M E 14

387 C F S L E M E E Ξ H L Y A L R 14

402 W K E L E M H S L A L Q N T L 14

407 M H S L A L Q N T L H E R T W 14

424 E K N L M Q Q E L R S L K Q N 14

436 K Q N I F L F Y V K L R w L L 14

443 Y V K L R W L L K ,H W R Q G K 14

475 L S R L G E L G V Q G G H Q A 14

478 L G E L G V Q G G H Q A D G P 14

480 E L G V Q G G H Q A D G P D H 14

510 H^'S R V Q I G D H S L R L Q T 14

512 R V Q I G D H S L R L Q T A D 14

532 K Q V V E N Q Q L F S A F K A 14

545 K A L L E D F R A E L R E D E 14

562 R L R L Q Q Q Y A S D K A A W 14

575 A W D V E W A V L K C R L E Q 14

579 E W A V L K c R L E Q L E E K 14 '

584 K C R L E Q L E E K T E N K L 14

626 H S L V M D L R W Q I H H S E 14

629 V M D L R W Q I H H s E K N W 14

633 R W Q I H H S E K N w N R E.K 14

685 G S F L C D Q K D G N V R P F 14 TABLE XXV - 185P2C9 v2 HLA- DRB1*0401 (DR4DW4 ) 15 - mers

Pos 1 2 3 9 0 1 Seq. ID

4 5 6 7 8 2 3 4 5 score Num.

702 Q G S L R M P R P V A M W P C 14

708 P R P V A M W P c A D A D s I 14

710 P V A M W P c A D A D S I P F 14

726 D R P L S K L K Ξ S D R c s A 14

741 S E N L Y L D A L S L D D E P 14

743 N L Y L D A L S L D D E P E E 14

746 L D A L S L D D E P E Ξ P P A 14

748 A L S L D D E P E E P P A H R 14

794 F Q R L M D I S P F L P E K G 14

797 L M D I S P F L P E K G L P S 14

806 E K G L P S T S s K Ξ D V T P 14

815 K E D V T P P L s P D D L K Y 14

827 L K Y I E E F N K s W D Y T P 14

879 T T S V T M T T D T M T S P E 14

881 S V T M T T D T M T S P E H C 14

897 K Q P L R s H V L T E Q S G L 14

902 S H V L T E Q S G L R V L H S 14

920 V R R V D S I T A A G G E G P 14

923 V D S I T A A G G E G P F P T 14

975 R R P L D S P L C T S L G F A 14

979 D S P L C T S L G F A S P L H 14

992 L H s L E M S K N L S D D M K 14

1002 S D D M K E V A F S V R N A I 14

1009 A F S V R N A I C S G P G E L 14

1013 R N A I C S G P G E L Q V K D 14

1022 E L Q V K D M A C Q T N G S R 14

1025 V K D M A C Q T N G S R T M G 14

1035 S R T M G T Q T V Q T I s V G 14

1052 T E A L R G S G V T S s P H K 14

1076 A T P V S S P S R s L R s R Q 14

1092 A P A I E K V Q A K F E R T C 14

TABLE XXVI - 185P2C9 v2 HLA- DRB1*1101 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

315 L S T V T S V S R D s P I G N 28 324 D S P I G N L G K E L G P D L 27 669 L W R I E Q L Q K E N S P R R 26 212 D S E Y L V T L K H E A Q R L 25 437 Q N I F L F Y V K L R W L L K 22 648 V E L L D R L D R D R Q E W E 22 791 M S E F Q R L M D I S P F L P 22 44 E E E A Ξ L L R R s I s E I E 21 58 E D H N R Q L T H E L s K F K 21 701 H Q G S L R M P R P V A M W P 21 719 A D S I P F E D R P L S K L K 21 776 E N H K G N L Q R A V S V S S 21 991 P L H s L E M S K N L S D D M 21 998 S K N L S D D M K E V A F S V 21 111 S G K V L K L Q H E N H A L L 20 122 H A L L S N I Q R C D L A A H 20 162 E S R L P Q P K R E G P V G G 20 213 S E Y L V T L K H Ξ A Q R L E 20 288 L Q A F L Q Q V N R I G D G L 20 289 Q A F L Q Q V N R I G D G L S 20 299 G D G L S P L P H L T E S S S 20 439 I F L F Y V K L R W L L K H W 20 TABLE XXVI - 185P2C9 v2 HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

443 Y V K L R W L L K H W R Q G K 20 463 G E E F T E G E H P E T L S R 20 519 S L R L Q T A D R G Q P H K Q 20 645 R E K V E L L D R L D R D R Q 20 693 D G N V R P F P H Q G S L R M 20 729 L S K L K E S D R C s A S E N 20 856 R T E V G R A G H E D S T E P ^'20 907 E Q S G L R V L H S P P A V R 20 914 L H S P P A V R R V D S I T A 20 1089 R Q V A P A I E K V Q A K F E 20 292 L Q Q V N R I G D G L S P L P 19 311 S S S F L S T V T S V S R D S 19 395 E H L Y A L R W K Ξ L E M H S 19 499 G C G F P V G E H S P H S R V 19 577 D V E W A V L K C R L E Q L E 19 794 F Q R L M D I S P F L P E K G 19 908 Q S G L R V L H s P P A V R R 19 967 S R D Y V E G A R R P L D S P 19 985 S L G F A S P L H S L E M S K 19 200 E P C P T E L L K A R E D S E 18 237 T D S F L H D A G L R G G A P 18 302 L S P L P H L T Ξ S S S F L S 18 475 L S R L G E L σ V Q G G H Q A 18 548 L E D F R A E L R E D E R A R 18 658 R Q E W E R Q K K Ξ F L W R I 18 684 G G S F L C D Q K D G N V R P 18 836 S W D Y T P N R G H N G G G P 18 917 P P A V R R V D S I T A A G G 18 920 V R R V D S I T A A G G E G P 18 30 L E S S T E L R R H L Q F V E 17 68 L S K F K F E P P R E P G W L 17 272 Q L L G T I N A K M K A F K K 17 362 R L R A A R E L H R R A D G D 17 440 F L F Y V K L R W L L K H W R 17 445 K L R W L L K H W R Q G K Q M 17 931 G E G P F P T S R A R G S P G 17 932 E G P F P T s R A R G S P G D 17 956 L S R W P C T S P R H S R D Y 17 1007 E V A F S V R N A I C S G P G 17 78 E P G W L G E G A s P G A G G 16 181 E E M F E K T S G F G S G K P 16 187 T S G F G s G K P s E A S E P 16 354 Q G D E R E S L R L R A A R E 16 399 A L R W K E L E M H S L A L Q 16 409 S L A L Q N T L H E R T W S D 16 411 A L Q N T L H E R T W S D E K 16 444 V K L R w L L K H w R Q G K Q 16 537 N Q Q L F S A F K A L L E D F 16 541 F S A F K A L L E D F R A E L 16 552 R A E L R E D E R A R L R L Q 16 576 W D V Ξ W A V L K C R L E Q L 16 605 A E S K G A L K K E R E V H Q 16 612 K K E R E V H Q K L L A D S H 16 689 C D Q K D G N V R P F P H Q G 16 799 D I S P F L P E K G L P S T S 16 800 I S P F L P E K G L P S T s S 16 830 I E E F N K S W D Y T P N R G 16 853 W A D R T E V G R A G H E D S 16 TABLE XXVI - 185P2C9 v2 HLA- DRB1*1101 15 - mers

Seq . ID

Pos . 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

886 T D T M T S P E H C Q K Q P L 16

933 G P F P T S R A R G S P G D T 16

1005 M K E V A F S V R N A I C S G 16

1018 S G P G E L Q V K D A C Q T 16

1028 M A C Q T N G S R T M G T Q T 16

1048 V G L Q T E A L R G S G V T S 16

1057 G S G V T S S P H K C L T P K 16

43 V E E E A E L L R R S I S E I 15

54 I S E I E D H N R Q L T H E L 15

62 R Q L T H E L S K F K F E P P 15

69 S K F K F Ξ P P R Ξ P G W L G 15

114 V L K L Q H E N H A L L S N I 15

161 E E S R L P Q P K R E G P V G 15

220 K H E A Q R L E R T V E R L I 15

224 Q R L E R T V E R L I T D T D 15

278 N A K M K A F K K E L Q A F L 15

305 L P H L T E S S S F L S T V T 15

363 L R A A R E L H R R A D G D T 15

372 R A D G D T G S H G L G G Q T 15

400 L R W K Ξ L E M H S L A L Q N 15

425 K N L M Q Q E L R S L K Q N I 15

510 H S R V Q I G D H S L R L Q T 15

618 H Q K L L A D S H S L V M D L 15

674 Q L Q K E N S P R R G G S F L 15

723 P F E D R P L S K L K E S D R 15

968 R D Y V E G A R R P L D S P L 15

1076 A T P V S S P S R S L R S R Q 15

1081 S P S R S L R S R Q V A P A I 15 llll Y G S P K L Q R K P L P K A D 15

2 E D T R G Q Q E R E G P G R D 14

7 Q Q E R E G P G R D H A P S I 14

9 E R E G P G R D H A P S I P T 14

64 L T H E L S K F K F E P P R E 14

93 G A P L Q E E L K S A R L Q I 14

108 S E L S G K V L K L Q H E N H 14

130 R C D L A A H L G L R A P S P 14

132 D L A A H L G L R A P S P R D 14

147 S D A E S D A G K K E S D G E 14

178 S D S E E M F E K T S G F G S 14

202 C P T E L L K A R E D S E Y L 14

234 I T D T D S F L H D A G L R G 14

239 s F L H D A G L R G G A P L P 14

274 L G T I N A K M K A F K K E L 14

335 G P D L Q S R L K E Q-L E W Q 14

364 R A A R E L H R R A D G D T G 14

388 F S L E M E E E H L Y A L R W 14

438 N I F L F Y V K L R W L L K H 14

446 L R W L L K H W R Q G K Q M E 14

469 G E H P ■E T L S R L G E L G V 14

490 D G P D H D S D R G C G F P V 14

517 D H S L R L Q T A D R G Q P H 14

542 S A F K A L L E D F R A E L R 14

544 F K A L L E D F R A E L R E D 14

625 S H S L V M D L R W Q I H H S 14

629^' V M D L R W Q i H H S E K N W 14

630 M D L R W Q I H H S E K N W N ^' 14

633 R W Q I H H s E K N W N R E K 14

663 R Q K K E F L W R I E Q L Q K 14 TABLE XXVI - 185P2C9 v2 HLA- DRB1*1101 15 - mers

Seq. ID

Pos • 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

675 Q K E N S P R R G G S F L C 14

699 F P H Q G s L R M P R P V A M 14

704 S L R M P R P V A M W P C A D 14

779 K G N L Q R A V S V S S M S E 14

788 V S S M S E F Q R L M D I S P 14

827 L K Y I E E F N K S W D Y T P 14

837 W D Y T P N R G H N G G G P D 14

881 S V T M T T D T M T S P E H C 14

923 V D S I T A A G G E G P F P T 14

940 A R G S P G D T K G G P P E P 14

950 G P P E P M L s R W P C T S P 14

980 S P L C T S L G F A S P L H S 14

1063 S P H K C L T P K A G G G A T 14

1065 H K C L T P K A G G G A T P V 14

1102 F E R T c C S P K Y G S P K L 14

1110 K Y G s P K L Q R K P L P K A 14

1115 K L Q R K- P L P K A D Q P N N 14

1127 P N N R P G N R H Q F P R K V 14

97 Q E E L K S A R L Q I s E L S 13

104 R L Q I s E L S G K V L K L Q 13

127 N I Q R c D L A A H L G L R A 13

131 C D L A A H L G L R A P S P R 13

136 H L G L R A P S P R D S D A E 13

180 S E E M F E K T S G F G S G K 13

238 D S F L H D A G L R G G A P L 13

295 V N R I G D G L S P L P H L T 13

340 S R L K E Q L E W Q L G P A Q 13

355 G D E R E S L R L R A A R E L 13

429 Q Q E L R S L K Q N I F L F Y 13

478 L G E L G V Q G G H Q A D G P 13

498 R G C G F P V G E H s P H S R 13

598 L G E L G s S A E S K G A L K 13

626 H S L V M D L R W Q I H H S E 13

797 L M D I S P F L P E K G L P S 13

812 T S S K E D V T P P L S P D D 13

849 G P D L W A D R T E V G R A G 13

899 P L R S H V L T E Q S G L R V 13

949 G G P P E P M L S R W P C T S 13

976 R P L D S P L C T S L G F A S 13

989 A S P L H s L E M S K N L S D 13

1002 S D D M K E V A F S V R N A I 13

1040 T Q T V Q T I S V G L Q T E A 13

1043 V Q T I S V G L Q T E A L R G 13

1047 S V G L Q T E A L R G S G V T 13

1113 S P K L Q R K P L P K A D Q P 13

TABLE XXIII 185P2C9 v3 HLA-

DRB *oιoι : 15 - mers

Pos 1 2 3 4 5 6 7 8 9 ' 0 1 , 2 3 4 5 score Seq. ID Num.

104 R L Q I S E L S G K V L K L Q 33 612 G S T V K T L K S L G L Q R L 33 918 Q S G L R V L H S P P A V R R 32 478 L G E L G V Q G G H Q A D G P 31 912 S H V L T E Q S G L R V L H S 31 181 E E M F E K T S G F G S G K P 30 927 P P A V R R V D S I T A A G G 30 942 E G P F P T s R A R G S P G D 29 39 H L Q F V E E E A E L L R R S 28 TABLE XXIII - 185P2C9 v3 :HLA-

DRB1*0101 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Seq. ID Num.

358 R E S L R L R A A R E L H R R 28

399 A L R W K E L E M H S L A L Q 28

804 F Q R L M D I S P F L P E K G 28

933 V D S I T A A G G E G P F P T 28

1175 S S L F N I I D H S P V V Q D 28

68 L S K F K F E P P R E P G W L 27

472 P E T L S R L G E L G V Q G G 27

603 T L G F T R L P A G S T V K T 27

1075 H K C L T P K A G G G A T P V 27

167 Q P K W E G P V G G E S D S E 26

246 L R G G A P L P G P G Ii Q G E 26

327 I G N L G K E L G P D L Q S R 26

429 Q Q E L R S L K Q N I F L F Y 26

615 V K T L K S L G L Q R L E L E 26

721 V A M W P C A D A D S I P F E 26

801 M S E F Q R L M D I S P F L P 26

810 I S P F L P E K G L P S T S S 26

1017 E V A F S V R N A I c S G P G 26

1059 G L Q T E A L R G S G V T s S 26

1254 G Y T L T E N V A R I L N K K 26

1274 L K Ξ E R R Q A A H G P P G L 26

136 H L G L R A P S P R D S D A E 25

394 E E H L Y A L R W K E L E M H 25

517 D H S L R L Q T A D R G Q P H 25

560 R A R L R L Q Q Q Y A S D K A 25

712 Q G S L R M P R P V A M W P C 25

883 D S S W Y L T T S V T M T T D 25

966 L S R W P C T S P R H S R D Y 25

1090 S S P S R S L R S R Q V A P A 25

1102 A P A I E K V Q A K F E R T C 25

79 P G W L G E G A S P G A G G G 24

112 G K V L K L Q H E N H A L L S 24

130 R C D L A A H L G L R A P s P 24

133 L A A H L G L R A P S P R D S 24

187 T S G F G S G K P S E A S E P 24

240 F L H D A G L R G G A P L P G 24

268 Q Q E P Q L L G T I N A K M K 24

292 L E Q V N R I G D G L S P L P 24

302 L S P L P H L T Ξ S S S F L s 24

311 S S S F L S T V T S V S R D s 24

424 E K N L M Q Q E L R S L K Q N 24

508 S P H S R V Q I G D H S L R L 24

696 R Q E W E R Q K K E F L W R I 24

714 S L R M P R P V A M W P C A D 24

750 A S E N L Y L D A L S L D D E 24

787 N H K G N L Q R A V S V S s M 24

813 F L P E K G L P S T S S K Ξ D 24

930 V R R V D S I T A A G G E G P 24

989 D S P L C T S L G F A S P L H 24

1004 S L E M S K N L S D D M K Ξ V 24

1246 K Q D L S A P P G Y T L T E N 24

1258 T E N V A R I L N K K L L E H 24

100 L K S A R L Q I S E L S G K V 23

271 P Q L L G T I N A K M K A F K 23

305 L P H L T E S S S F L S T V T 23

309 T E S S S F L S T V T S V S R 23

315 L S T V T S V S R D S P I G N 23

339 Q s R L K E Q L E W Q L G P A 23

438 N I F L F Y V K L R W L L K H 23 to to to . ^ " vo vD θo oo oo --J cn ιf* ιo , ₁ vo co co -o. c ιi*. ιf* M to M . [ ^ ^ ^ ^ vo vo vo oo ι cn cn ιπ ιn ιo ιo ιo to t H ._n - t-¹ - b-¹ I--

^ ^w l^J _ffi m ϊfr u ^ ° * ^{M o} ^ ^o ^ ^ ^ffl * * ° * ^{o m} n m ι ^ ϊ ^|l^{| |}Λ O I0 α) lIl U C. W l *> * l. l U Ϊ VO tO H H O O vo ~o -J cn i-j -ι m ιπ "' "ⁱ "n^l »^J*' H H __{1 m} ,_rv , „, ,., ,n ι__ M ^ul rf* oo σ> in rf* in to ~o VD to cn -J rf* to

a rt a a rt rt to O rt 3 a O a cn a Ω a H a a a 2 B 2 O H B KD tr a co a a a a a K H U⁾ f. 01 O t. a a to Ώ < a a rt Ω a a a a α a a < B 3 a a a rt cn O a a a cn o Ω rt a a a a a KD a Ώ a a α cn cn a Ω B Ω α Ω a a a D a a a a a > TJ rt < rt rt a rt a a a a rt < Ω a to Ω CO α a a cn H a a a a a TJ rt a 2 > •fl CO < B α <: TJ cn a rt •A a KD < a a rt a < Ώ a a rt a a a ιo rt H Ω n IT Ά a

KD o rt s: rt a rt < a a a cn a 2 B a to a cn a a Ω Ki a a a Ω a to a a a 2 3 a rt a a a a a 2 a H < a KD a H CO cn a 2 a a

< O a a ^ a D a <! co a a a a a cn a B < a-H CO < te a a σ a a KD a cn Ω cn a KD a to a D a Ω iO B Ω B cn B a a a < a Ω a 2 Ω a a a Ω H Ω rt a a a Ω α a s H a rt cn a rt a B rt a a i ^ C) P3 H a a a rt a cn a a a α a a Ω a 2 α a a B KD a α Ω rt ^ -. ?3 B a a a B a cn < > Ω < a < a a a 3 < H 3 3 n D < a KD Ω B KD KD a Ω H KD a a a a > cn KD KD tO Ω KD Ω a H a O KD Ώ a 3 to B H a

Lπ O H a a Ω D Ω KD H > a a i a H a a rt a a rt 2 Ω Pi a a α a a a a a H <! rt a 2 a cn < a Ω Ki o rt a a a H iO a a a a ιo Ω to D B Ω

^ Ω α to to Ω <! rt Ω a rt Ω a B a ω to CO rt CO rt a to rt 2 rt Ώ D a rt a a σ < to a cn a cn CO B to tT Ω to Ω cn rt a co to Ω cn rt rt a Ω KD cn VD α to ιθ cn a rt H O a a Ω a a KD B a cn a α K; IT a a a a > K a a to a a D a Ω a α < a a co B a a a a O Ω > a co a K a co Ω ω O ^■

Ω Ω a a cn co cn Ω a iO Ω a Ω Ω a > cn a 2 rt rt a a a a cn a a a a Ω cn > rt B rt o CO < Ω a cn to Ω to a rt a a a a a > rt a a co to H 3 a a a Ω cn to < a a cn a cn a cn a a a a a 2 a 2 to a a trj H H .H a ω TJ H H 2 cn a a to 2 cn to IT α a a σ a a j ; a a < cn a a a to cn a B to a a Ω Ω o Ω o a a a 2 a Ω Ω rt a a a a a a B Ω cn B a < a to cn a rt a 2 α cn te B tfl B Ω a cn > o Ω Ω a a iO a s a CO ⁰¹ ω a Ω a Ω a a a a cn a a σ to a a a cn o rt cn a a Ω 2 rt a KD rt a a n < Pi < a a B i ω a to > ?.3 a o a a te H to H Ω rt B a 2 a rf_* a Ω rt Ω a a a a a 2 α rt 3 a rt a cn cn a Ω a a a 2 a H ifl to a iO Ω a a KD H a a B 2 to B B B rt a a cn rt a H ω H 3 a a Ω

H H H H tO tO tO tO tO tO tO tO tO tO tO tO tO tO tO tO tO tO tO tO W tO tO tO tO tO tO tO tO M tO tO tO W tO W tO tO M VD VO VD VO O O O O O O O O O O O O O O H H H H H H H H H H H tO tO M tO W tO tO tO tO tO tO M W W tO tO tO tO tO tO tO LO lo ω ω io ω OO LO lO ^ o

TABLE XXIII - 185P2C9 v3 HLA- DRB1*0101 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Seq. ID Num.

475 L S R L G E L G V Q G G H Q A 19

541 F S A F K A L L E D F R A E L 19

552 R A E L R E D E R A R L R L Q 19

577 D V E W A V L K C R L E Q N C 19

604 L G F T R L P A G S T V K T L 19

678 E K N W N R E K V E L L D R L 19

731 S I P F E D R P L S K L K E S 19

846 S W D Y T P N R G H N G G G P 19

1057 S V G L Q T E A L R G S G V T 19

1120 K Y G S P K L Q R K P L P K A 19

1184 S P V V Q D P F Q K G L R A G 19

1223 R S P S P I G V G S E M C R E 19

1262 A R I L N K K L L E H A L K E 19

1263 R I L N K K L L E H A L K E Ξ 19

58 E D H N R Q L T H Ξ L S K F K 18

65 T H E L S K F K F E P P R Ξ P 18

75 P P R E P G W L G E G A S P G^' 18

77 R E P G w L G E G A S P G A G 18

83 G E G A s P G A G G G A P L Q 18

111 S G K V L K L Q H E N H A L L 18

123 A L L S N I Q R C D L A A H L 18

126 S N I Q R c D L A A H L G L R 18

131 C D L A A H L G L R A P S P R 18

162 E S R L P Q P K W E G P V G G 18

185 E K T S G F G S G K P S E A S 18

190 F G S G K P S E A S E P C P T 18

201 P C P T E L L K A R Ξ D s E Y 18

212 D S E Y L V T L K H E A Q R L 18

228 R T V E R L I T D T D S F L H 18

237 T D S F L H D A G L R G G A P 18

241 L H D A G L R G G A P L P G P 18

435 L K Q N I F L F Y V K L R W L 18

439 I F L F Y V K L R W L L K H W 18

443 Y V K L R W L L K H W R Q G K 18

461 E E G Ξ E F T E G E H P Ξ T L 18

481 L G V Q G G H Q A D G P D H D 18

491 G P D H D s D R G C G F P V G 18

535 V E N Q Q L F S A F K A L L E 18

538 Q Q L F s A F K A L L E D F R 18

542 S A F K A L L E D F R A E L R 18

566 Q Q Q Y A S D K A A W D V E W 18

573 K A A W D V E W A V L K C R L 18

598 N I E E E T L G F T R L P A G 18

637 G E L G S s A E S K G A L K K 18

653 R E V H Q K L L A D S H S L V 18

654 Ξ V H Q K L L A D S H S L V M 18

663 S H S L V M D L R W Q I H H S 18

736 D^" R P L s K L K E s D R C S A 18

774 E R E F R N R L P E E E Ξ N H 18

891 S V T M T T D T M T S P E H C 18

953 S P G D T K G G P P Ξ P M L S 18

976 H S R D Y V E G A R R P L D S 18

990 S P L C T s L G F A S P L H S 18

999 A S P L H s L E M S K N L S D 18

1011 L S D D M K E V A F S V R N A 18

1067 G S G V T s S P H K C L T P K 18

1077 C L T P K A G G G A T P V S S 18

1078 L T P K A G G G A T P V S S P 18

1093 S R S L R s R Q V A P A I E K 18

a a a to a a rt cn a to J a a a a cn H a a α KD 2 2 o a Ω α a a a Ώ a D a rt te cn α D a Ω Ω a a a a < TJ a o s a a KD KD a KD rt a a a H rt a a a co α a B a a a a rt J Ω t' n H a α co α to i-3 a o a < KD B 10 te B Ω Ω a KD a a co o a Ω Ω to B a cn « Ω to o a 3 rt a a a a a cn a a a a a Ω B H Ω a a D Ω to a a cn te a H KD KD > B a Ki B Ω B Ω cn te a a a a H rt to a a a rq Ω a Ω Ω to σ to a a to < Ω a a H < H < H a a B a α Ω KD a a to α a H T) h_ a a a B Ω a a a M a a te TJ ω a rt B a a a a to Ω Ώ a < K; cn rf* a a a α cn te Ω a co a a a ; B B a B KD rt hi < J Ω 3 a a a Ω B Ώ H a a > IT TJ a a a < 3 a a a Ω a Ω a B to rt Ω Ω B a a rt a cn TJ in H

* a cn a Ω a a T a B Ά 3 KD a a Ba a a Ω •A a a Ω o to a Ω a a « Ω KD a H a rt Ά a to a te Ω 2 a a a to o Ω CO cn a to rt a a cn o

H

Ω a 2 Ω te Ω α 3 cn B 2 Ω a <; a a Ώ a a KD a a a a rt a a a to a s rt a Ω rt rt a a to te KD Ω te a a a a a KD rt Ω a TJ co Ω rt cn Kj O l_π H 1 o rt a a Ώ Ώ o α o α a to o a rt a a a KD a < a to 3 Ώ Ώ a Ω a < to pi H a Ώ α < a a a a B Ω Ω Ω cn a a a B to B H cn a cn H to Ώ 00 H cn Ω Ω Ω a Ω IT H a a Ω a a KD B rt rt KD a Ω Ω Ω KD Ω cn a a Ω a > a Ω rt a ri Ω a B Ω Ω B a a a Ώ a B Ω Ω a Ω a a 2 cn vo in 0 i a rt rt a a a a co Ω a t a a aaa B a b KD a a a a a a a to H TJ α a B CO a o a > Ω Ω a cn a α a TJ rt < H a to σ te o ' t

Ω cn cn Ά o o K! TJ ω a a 3 α a a Ω a KD a B KD B rt Ω a B H a a n cn a > Ώ B H a a to t*: H CO CO Ώ Ω a Ω Ω to a Ω a a H 3 a a a cn a H a te cn J a a a a cn < O "fl 3 a Ω a to Ω a Ώ a a 3 a a •fl H KD a cn cn cn a a co a cn H a a a < co < Ti co a co a t to Ω a rt rt 2 a a to t*! α a o > Ω rt to rt to < TJ rt B a a Ώ a a Ω α KD a a Ώ a ^ a Ώ a a > a a a B to a Ω s a a Ω cn te l

CO H KD LO co a a cn B to B a a a co TJ a α a a Ώ a a a a a cn to Ω a o a Ώ a a x a a to a a a a Ώ a H B cn a B KD 3 cn TJ B to hi ta rf_* a ω a TJ O a B a a H <J KD W a a cn a en Ω a a a KD a a o a a a 3 Ω O rt B Ω TJ cn a B B TJ B H cn α a o Ω B H TJ a rt a in I

_{H H H} μ_ _H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H "

-0. ,4 -J -J 1 -0 ^ «j ^ ^ ι ^ ^ ^ _ ^ ^ ^ -^ ^ ^ j ^ ^ -j ι ^ ^ ^ ^ ^ ^ ^ _ v_ ^ -o -j j ^ j ^ ^ j ^ ^ ι _o _o ω eo _o _o (n _o co !q

00 rt

TABLE XXIII - 185P2C9 v3 :HLA-

DRB1*0101 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Seq. ID Num.

991 P L C T S L G F A S P L H S L 17

1015 M K E V A F S V R N A I C S G 17

1027 C S G P G E I" Q V K D M A c Q 17

1049 G T Q T V Q T I S V G L Q T E 17

1053 V Q T I S V G L Q T E A L R G 17

1071 T S s P H K C L T P K A G G G 17

1123 S P K L Q R K P L P K A D Q P 17

1126 Q R K P L P K A D Q P N N R 17

1134 A D Q P N N R T S P G M A Q K 17

1142 S P G M A Q K G Y s E S A W A 17

1152 E S A W A R S T T T R E S P V 17

1153 S A W A R S T T T R E S P V H 17

1176 S L F N I I D H s P V V Q D P 17

1189 D P F Q K G L R A G S R S R S 17

1190 P F Q K G L R A G S R S R S A 17

1209 E L G P G Q E T G T N S R G R 17

1231 G S E M C ,R E E G G E G T P V 17

1241 E G T P V K Q D L S A P P G Y 17

1271 E H A L K E E R R Q A A H G P 17

1277 E R R Q A A H G P P G L H S D 17

10 R E G P G R D H A P S I P T S 16

11 E G P G R D H A P S I P T S P 16

18 A P s I P T S P F G D S L E S 16

25 P F G D S L E S s T E L R R H 16

37 R R H L Q F V E E Ξ A E L L R 16

46 E A E L L R R S I S E I E D H 16

47 A E L L R R S I s E I E D H N 16

61 N R Q L T H E L s K F K F E P 16

99 E L K S A R L Q I S E L S G K 16

114 V L K L Q H E N H A L L S N I 16

115 L K L Q H E N H A L L S N I Q 16

127 N I Q R C D L A A H L G L R A 16

155 K K E S D G E E S R L P Q P K 16

184 F E K T S G F G S G K P S E A 16

230 V E R L I T D T D S F L H D A 16

243 D A G L R G G A P L P G P G L 16

251 P L P G P G L Q G E E E Q G E 16

289 Q. A F L E Q V N R I G D G L S 16

299 G D G L S P L P H L T E S S S 16

303 S P L P H L T E S S S F L S T 16

314 F L S T V T S V S R D S P I G 16

318 V T s V S R D S P I G N L G K 16

328 G N L G K E L G P D L Q S R L 16

331 G K E L G P D L Q S R L K E Q 16

335 G P D L Q S R L K E Q L E W Q 16

347 E W Q L G P A R G D E R E S L 16

357 E R E S L R L R A A R E L H R 16

389 S L E M E E E H L Y A L R W K 16

391 E M E E E H L Y A L R W K E L 16

404 E L E M H S L A L Q N T L H E 16

431 E L R S L K Q N I F L F Y V K 16

493 D H D S D R G C G F P V G E H 16

509 P H S R V Q I G D H S L R L Q 16

514. Q I G D H S L R L Q T A D R G 16

519 S L R L Q T A D R G Q P H K Q 16

537 N Q Q L F S A F K A L L E D F 16

544. F K A L L E D F R A E L R E D 16

545 K A L L E D F R A E L R E D E 16

559 E R A R L R Q Q Q Y A S D K 16 TABLE XXI II - 185P2C9 V3 HLA- DRB1*0101 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Seq. ID Num.

567 Q Q Y A S D K A A W D V E W A 16

584 K C R L E Q N C C G Y P R I N 16

591 C C G Y P R I N I E E E T L G 16

602 E T L G F T R L P A G S T V K 16

617 T L K S L G L Q R L E L E E K 16

632 T E N K L G E L G S S A E S K 16

652 E R E V H Q K L L A D S H S L 16

656 H Q K L L A D S H S L V M D L 16

664 H S L V M D L R W Q I H H S E 16

667 V M D L R W Q I H H S E K N W 16

686 V E L L D R L D R D R Q E W E 16

709 R I E Q G S L R M P R P V A M 16

710 I E Q G S L R M P R P V A M W 16

720 P V A M W P C A D A D S I P F 16

743 K E S D R C S A S E N L Y L D 16

748 C S A S E N L Y L D A L s L D 16

752 E N L Y L D A L S L D D E P E 16

753 N L Y L D A L S L D D E P E E 16

758 A L S L D D E P E E P P A H R 16

762 D D E P E E P P A H R P E R E 16

786 E N H K G N L Q R A V S V S S^' 16

790 G N L Q R A V S V S S M S E F 16

793 Q R A V S V S S M S E F Q R L 16

812 P F L P E K G L P S T S S K Ξ 16

834 P D D L K Y I E E F N K S W D 16

856 N G G G P D L W A D R T E V G 16

915 L T E Q S G L R V L H S P P A 16

919 S G L R V L H S P P A V R R V 16

921 L R V L H S P P A V R R V D S 16

951 R G S P G D T K G G P P E P M 16

982 E G A R R P L D S P L C T S L 16

1000 S P L H S L E M S K N L S D D 16

1008 S K N L S D D M K E V A F S V 16

1023 R N A I C S G P G E L Q V K D 16

1034 Q V K D M A C Q T N G S R T M 16

1037 D M A C Q T N G s R T M G T Q 16

1061 Q T E A L R G S G V T S S P H 16

1072 S S P H K C L T P K A G G G A 16

1085 G A T P V S S P s R S L R S R 16

1107 K V Q A K F E R T C C S P K Y 16

1121 Y G S P K L Q R K P L P K A D 16

1143 P G M A Q K G Y s E S A w A R 16

1171 N D G L S S L F N I I D H S P 16

1217 G T N S R G R S P S P I G V G 16

1243 T P V K Q D L S A P P G Y T L 16

1251 A P P G Y T L T E N V A R I L 16

1279 R Q A A H G P P G L H S D s H 16

1288 L H S D S H S L G D T A E P G 16

TABLE XXIV - 185P2C9 v3 HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

544 F K A L L E D F R A E L R E D 38 552 R A E L R E D E R A R L R L Q 38 331 G K E L G P D L Q S R L K E Q 36 230 V E R L I T D T D S F L H D A 29 663 S H S L V M D L R W Q I H H S 29 429 Q Q E L R S L K Q N I F L F Y 28 TABLE XXIV - 185P2C9 v3 HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

339 Q S R L K E Q L E w Q L G P A 27 387 C F S L E M E E E H L Y A L R 27 537 N Q Q L F S A F K A L L E D F 27 625 R E L E Ξ K T E N K L G E L 27 756 L D A L S L D D E P E Ξ P P A 27

53 S I S E I E D H N R Q L T H E 26

173 P V G G E s D S E E M F E K T 26

335 G P D L Q s R L K Ξ Q L E W Q 26

350 L G P A R G D E R E S L R L R 26

689 L D R L D R D R Q E W E R Q K 26

1008 S K N L S D D M K E V A F S V 26

1258 T E V A R I L N K K L L E H 26

281 M K A F K K E L Q A F L E Q V 25

704 K E F L W R I Ξ Q G S L R M P 25

122 H L L s N I Q R C D L A A H 24

439 I F L F Y V K L R W L L K H W 24

656 H Q K L L A D S H S L V M D L 24 .

1163 E S P V H T T I N D G L S S L 24

1183 H S P V V Q D P F Q K G L R A 24

1184 S P V V Q D P F Q K G L R A G 24

1242 G P V K Q D L S A P P G Y T 23

1285 P P G L H S D S H S L G D T A 23

615 V K T L K s L G L Q R L E L E 22

751 S E N L Y L D A L S L D D E P 22

126 S N I Q R C D L A A H L σ L R 21

237 T D S F L H D A G L R G G A P 21

318 V T S V S R D S P I G N L G K 21

612 G S T V K T L K s L G L Q R L 21

803 E F Q R L M D I s P F L P E K 21

829 T P P L S P D D L K Y I Ξ E F 21

891 S V T M T T D T M T S P E H C 21

1067 G S G V T S S P H K c L T P K 21

39 H L Q F V E E E A E L L R R S 20 104 R L Q I s E L S G K V L K Q 20 114 V L K L Q H E N H A L L S N I 20 204 T E L K A R E D S Ξ Y L V T 20 206 L L K A R E D S E Y L V T L K 20 270 E P Q L L G T I N A K M K A F 20 294 Q V N R I G D G L S P L P H L 20 358 R E S L R L R A A R E L H R R 20 389 S L E M E E E H L Y A L R W K 20 417 H E R T W S D E K N L M Q Q E 20 432 L R S L K Q N I F L F Y V K L 20 511 S R V Q I G D H s L R L Q T A 20 580 W V L K C R L E Q N C C G Y 20 821 S T S S K E D V T P P L S P D 20 859 G P D L w A D R T E V G R A G 20 870 G R A G H E D S T E P F P D S 20 877 S T E P F P D S s W Y L T T s 20 985 R R P L D S P L C T S L G F A 20 1166 V H T T I N D G L S S L F N I 20 1167 H T T I N D G L s S L F N I I 20 1254 G Y T L T E N V A R I L N K K 20 1271 E H A L K E E R R Q A A H G P 20

40 L Q F V E E E A E L L R R S I 19

93 G A P L Q E E L K S A R L Q I 19 112 G K V L K L Q H Ξ N H A L L S 19 121 N H A L L S N I Q R C D L A A 19 TABLE XXIV - 185P2C9 v3 HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

214 E Y L V T L K H E A Q R L E R 19

278 N A K M K A F K K E L Q A F L 19

315 L S T V T S V S R D S P I G N 19

327 I G N L G K E L G P D L Q S R 19

424 E K N L M Q Q E L R S L K Q N 19

438 N I F L F Y V K L R W L L K H 19

446 L R W L L K H w R Q G K Q M E 19

491 G P D H D S D R G C G F P V G 19

531 H K Q V V E N Q Q L F S A F K 19

560 R A R L R L Q Q Q Y A S D K A 19

778 R N R L P E Ξ E Ξ N H K G N L 19

793 Q R A V S V S S M S E F Q R L 19

834 P D D L K Y I Ξ E F N K S W D 19

1002 L H S L E M s K N L S D D M K 19

1055 T I S V G L Q T E A L R G S G 19

1086 A T P V S S P S R S L R S R Q^' 19

1246 K Q D L S A P P G Y T L T E N 19

27 G D S L Ξ S s T E L R R H L Q 18

33 S T E L R R H L Q F V E E E A 18

37 R R H L Q F V E E E A Ξ L L R 18

54 I S E I E D H N R Q L T H E L 18

113 K V L K L Q H E N H A L L S N 18

153 A G K K E S D G E E S R L P Q 18

203 P T E L L K A R E D S E Y L V 18

216 L V T L K H E A Q R L Ξ R T V 18

254 G P G L Q G E E E Q G E G D Q 18

285 K K E L Q A F L Ξ Q V N R I G 18

324 D s P I G N L G K E L G P D L 18

360 S L R L R A A R E L H R R A D 18

379 S H G L G G Q T C F S L E M E 18

386 T C F S L E M Ξ E E H L Y A L 18

394 E E H L Y A L R W K Ξ L E M H 18

425 K N L M Q Q E L R S L K Q N I 18

443 Y V K L R W L L K H W R Q G K 18

447 R W L L K H W R Q G K Q M E E 18

456 G K Q M E E E G E E F T Ξ G E 18

472 P E T L S R L G E L G V Q G G 18

558 D E R A R L R L Q Q Q Y A S D 18

594 Y P R I N I E E E T L G F T R 18

623 L Q R L E L E E K T E N K L G 18

636 L G E L G S S A E S K G A L K 18

646 K G A L K K E R E V H Q K L L 18

671 R W Q I H H S E K N W N R E K 18

683 R E K V E L L D R L D R D R Q 18

731 S I P F E D R P L S K L K E S 18

739 L s K L K E S D R C S A S E N 18

744 E s D R C S A S E N L Y L D A 18

795 A V S V S S M S Ξ F Q R L M D 18

798 V s S M S E F Q R L M D I S P 18

1015 M K E V A F S V R N A I c S G 18

1101 V A P A I E K V Q A K F E R T 18

1192 Q K G L R A G S R S R S A E P 18

TABLE XXV - 185P2C9 v3 HLA- DRB1*0401 (DR4Dw4) 15 - mers

Seq. ID os 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num. 81 M K A F K K E L Q A F L E Q V 28 TABLE XXV - 185P2C9 v3 HLA- DRB1*0401 (DR4Dw4 ) 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

418 E R T W S D E K N L M Q Q E L 28 1175 S S F N I I D H S P V V Q D 28 61 N R Q L T H E L S K F K F E P 26 114 V L K L Q H E N H A L L S N I 26 122 H A L L S N I Q R C D L A A H 26 213 S E Y L V T L K H E A Q R L E 26 331 G K E L G P D Q S R L K E Q 26 425 K N L M Q Q E L R S L K Q N I 26 544 F K A L L E D F R A E L R E D 26 552 R A E L R E D E R A R L R L Q 26 612 G S T V K T L K S L G L Q R L 26 656 H Q K L L A D S H S L V M D L 26 663 S H S L V M D L R W Q I H H S 26 686 V E L L D R L D R D R Q E W E 26 704 K E F L W R I E Q G S L R M P 26 729 A D S I P F Ξ D R P L S K L K 26 789 K G N L Q R A V S V S S M S E 26 866 R T E V G R A G H E D S T E P 26 918 Q S G L R V L H S P P A V R R 26 927 P P A V R R V D S I T A A G G 26 1128 R K P L P K A D Q P N N R T S 26 1258 T E N V A R I L N K K L L E H 26 1271 E H A L K E E R R Q A A H G P 26 39 H L Q F V E E E A E L L R R S 22 311 S S S F L S T V T s V S R D S 22 399 A L R W K Ξ L E M H S L A L Q 22 440 F L F Y V K L R W L L K H W R 22 463 G E E F T E G E H P Ξ T L S R 22 548 L E D F R A E L R E D E R A R 22 573 K A A W D V E W A V L K C R L 22 603 T L G F T R L P A G S T V K T 22 669 D L R W Q I H H S E K N W N R 22 678 E K N w N R E K V E L L D R L 22 721 V A M w P C A D A D S I P F E 22 752 E N L Y L D A L S L D D E P E 22 810 I S P F L P E K G L P S T S S 22 836 D L K Y I E E F N K s W D Y T 22 883 D S S w Y L T T S V T M T T D 22 977 S R D Y V E G A R R P L D s P 22 1188 Q D P F Q K G L R A G S R s R 22 54 I S E I E D H N R Q L T H E L 20 79 P G W L G E G A S P G A G G G 20 97 Q E E L K S A R L Q I S E L S 20 104 R L Q I S E L S G K V L K L Q 20 111 S G K V L K L Q H E N H A L L 20 216 L V T L K H E A Q R L E R T V 20 227 E R T V E R L I T D T D S F L 20 230 V E R L I T D T D S F L H D A 20 270 E P Q L L G T I N A K M K A F 20 271 P Q L L G T I N A K M K A F K 20 278 N A K M K A F K K E L Q A F L 20 285 K K E L Q A F L E Q V N R I G 20 289 Q A F L E Q V N R I G D G L S 20 292 L E Q V N R I G D G L S P L P 20 299 G D G L S P L P^'H L T E S S S 20 302 L S P L P H L T E S S S F L S ^'20 305 L P H L T-E S S S F L S T V T 20. 312 S S F L S T V T S V S R D s P 20 TABLE XXV - 185P2C9 v3 HLA- DRB1*0401 (DR4Dw4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

315 L S T V T S V S R D S P I G N 20 318 V T S V s R D S P I G N L G K 20 327 I G N L G K E L G P D L Q S R 20 389 S L E M E E E H L Y A L R W K 20 394 Ξ E H L Y A L R W K E L E M H 20 404 E L E M H S L A L Q N T L H E 20 409 S L A L Q N T L H E R T W S D 20 429 Q Q E L R S L K Q N I F L F Y 20 438 N I F L F Y V K L R w L L K H 20 446 L R W L L K H W R Q G K Q M E 20 447 R W L L K H w^' R Q G K Q M E E 20 456 G K Q M E E E G E E F T E G E 20 472 P E T L S R L G E L G V Q G G 20 519 S L R L Q T A D R G Q P H K Q 20 531 H K Q V V E N Q Q L F S A F K 20 537 N Q Q L F S A F K A L L E D F 20 560 R A R L R L Q Q Q Y A S D K A 20 596 R I I E E E T L G F T R L P 20 606 F T R L P A G s T V K T L ,K S 20 615 V K T L K S L G L Q R L E L Ξ 20 620 S L G L Q R L E L E Ξ K T E N 20 623 L Q R L E L E E K T E N K L G 20 633 E N K L G E L G S S A E S K G 20 646 K G A L K K E R E V H Q K L L 20 657 Q K L L A D S H S L V M D L R 20 683 R E K V E L L D R L D R D R Q 20 689 L D R L D R D R Q E W E R Q K 20 739 L S K L K E S D R C S A S E N 20 778 R N R L P E E E E N H K G N L 20 798 V S S M S E F Q R L M D I S P 20 829 T P P L S P D D L K Y I E E F 20 834 P D D L K Y I E E F N K S W D 20 859 G P D L W A D R T E V G R A G 20 885 S W Y L T T S V T M T T D T M 20 896 T D T M T s P E H C Q K Q P L 20 911 R S H V L T E Q S G L R V L H 20 978 R D Y V E G A R R P L D S P L 20 993 C T S L G F A s P L H S L E M 20 999 A S P L H s L E M S K N^' L S D 20 1004 S L E M S K N L S D D M K E V 20 1008 S K N L S D D M K E V A F S V 20 1015 M K E V A F S V R N A I C S G 20 1050 T Q T V Q T I s V G L Q T E A ^' 20 1053 V Q T I S V G L Q T E A L R G 20 1057 S V G L Q T E A L R G S G V T 20 1067 G S G V T s S P H K C L T P K 20 1098 S R Q V A P A I E K V Q A K F 20 1163 E S P V H T T I N D G L S S L 20 1171 N D G L S s L F N I I D H S P 20 1184 S P V V Q D P F Q K G L R A G 20 1192 Q K G L R A G S R S R S A E P 20 1242 G T P V K Q D L S A P P G Y T 20 1285 P P G L H S D S H S L G D T A 20

12 G P G R D H A P S I P T S P F 18

25 P F G D S L E S S T E L R R H 18

29 S L E S S T E L R R H L Q F V 18

30 L E S S T E L R R H L Q F V E 18

43 V E E E A E L L R R S I S E I 18 TABLE XXV - 185P2C9 v3 HLA- DRB1* 0401 (DR4D 4 ) 15 - mers

Seq. ID

POS 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

44 E E E A E L L R R s I S E I E 18 52 R S I S E I E D H N R Q L T H 18 53 S I S E I E D H N R Q L T H E 18 58 E D H N R Q L T H E L S K F K 18 94 A P L Q E E L K S A R L Q I S 18 103 A R L Q I S E L S G K V L K L 18 118 Q H E N H A L L S N I Q R c D 18 128 I Q R C D L A A H L G L R A P 18 156 K E S D G E Ξ S R L P Q P K W 18 205 E L L K A R Ξ D S E Y L V T L 18 224 Q R L E R T V E R L I T D T D 18 261 E E Q G E G D Q Q E P Q L L G 18 268 Q Q E P Q L L G T I N A K M K 18 304 P L P H L T E S s s F L S T V 18 308 L T E S S S F L s T V T S V s 18 309 T E S S S F L S T V T S V S R 18 321 V S R D S P I G N L G K E L G 18 332 K E L G P D L Q S R L K E Q L 18 350 L G P A R G D E R E S L R L R 18 362 R L R A A R E L H R R A D G D 18 388 F S L Ξ M E E E H L Y A L R W 18 400 L R W K E L E M H S L A L Q N 18 405 L E M H S L A L Q N T L H E R 18 421 W S D E K N L M Q Q Ξ L R S L 18 426 N L M Q Q E L R S L K Q N I F 18 444 V K L R w L L K H W R Q G K Q 18 500 C G F P V G E H S P H S R V Q 18 511 S R V Q I G D H S L R L Q T A 18 529 Q P H K Q V V E N Q Q L F s A 18 534 V V E N Q Q Ii F S A F K A L L 18 559 E R A R L R L Q Q Q Y A S D K 18 600 E E E T L G F T R L P A G S T 18 605 G F T R L P A G S T V K T L K 18 609 L P A G S T V K T L K S L G L 18 617 T L K S L G L Q R L E L E E K 18 624 Q R L E L E E K T E N K L G E 18 637 G E G S S A Ξ s K G A L K K 18 648 A L K K E R'E V H Q K L L A D 18 649 L K K Ξ R E V H Q K L L A D S 18 655 V H Q K L L A D s H S L V M D 18 668 M D L R W Q I H H S E K N W N 18 701 R Q K K E F L W R I E Q G S L 18 770 A H R P E R E F R N R L P E E 18 780 R L P E E Ξ E N H K G N L Q R 18 786 E N H K G N L Q R A V S V S S 18 797 S V S S M S Ξ F Q R L M D I S 18 803 E F Q R L M D I S P F L P E K 18 813 F P E K G L P S T S S K E D 18 821 S T S S K E D V T P P L S P D 18 858 G G P D L W A D R T E V G R A 18 877 S T E P F P D S S W Y L T T S 18 882 P D S S W Y L T T S V T M T T 18 904 H C Q K Q P L R s H V L T E Q 18 908 Q P L R S H V L T E Q S G L R 18 924 L H S P P A V R R V D S I T A 18 939 A G G E G P F P T S R A R G S 18 996 L G F A S P L H S L E M S K N 18 1000 S P L H S L E M S K N L S D D 18 TABLE XXV - 185P2C9 v3 HLA- DRB1*0401 (DR4DW4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1013 D D M K E VA N A I C 18

1047 T M G T Q T V V G Q 18

1054 Q T G E A L R G S 18

1090 S S S R Q V A P A 18

1107 K V F S P K Y 18

1120 K Y K K A 18

1150 Y W A E S 18

1160 T R E P V 18

1168 T G 18

1176 S I 18

1199 S A 18

1235 C E E G 18

1239 G E G P 18

1255 Y L T E N V N K 18

1264 I N K K L L E H K E 18

1270 L K E E R 18

439 I V K H W 17

23 T D S L R 16

68 L F 16

78 E G 16

167 Q K W G 16

181 E M F E K 16

187 T G S 16

212 D V H E 16

237 T H D L R 16

288 L E Q V N 16

385 Q L E M E 16

450 L H W Q G K Q 16

499 G G F V G E H R V 16

538 Q A F F R 16

541 F E L 16

566 Q D V E W 16

577 D A V E Q 16

591 C P R 16

696 R E W K K 16

705 E Q G 16

801 M 16

844 N K H N G G 16

878 T E T T S V 16

884 S 16

942 E 16

966 L R W 16

995 S G F Ξ M 16

1109 Q K F E R K Y 16

1118 S K Y G S 16

1147 Q K G Y 16

1152 E S A W 16

1252 P G Y E N 16

18 A P F 14

33 S E E 14

37 R E L 14

40 L R R 14

47 A E D H N 14

51 R H N R Q L T 14

65 T E P P R E P 14

93 G S A R L Q I 14

102 S S G K V L K 14

112 G H E N H A L L S 14 TABLE XXV - 185P2C9 v3 HLA- DRB1*0401 (DR4DW4 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

121 H L L S N I Q R C D L A A 14 130 R C D L A A H L G L R A P S P 14 136 H L G L R A P Ξ P R D S D A E 14 171 E G P V G G E S D S E E M F E 14 203 P T E L L K A R E D S E Y L V 14 223 A Q R L E R T V E R L I T D T 14 231 E R L I T D T D S F L H D A G 14 238 D S F L H D A G L R G G A P L 14 254 G P G L Q G E E E Q G E G D Q 14 295 V N R I G D G L S P L P H L T 14 324 D S P I G N L G K E L G P D L 14 339 Q S R L K E Q L E W Q L G P A 14 347 E W Q L G P A R G D E R E S L 14 360 S L R L R A A R E L H R R A D 14 379 S H G L G G Q T C F S L E M Ξ 14 387 C F S L E M E E Ξ H L Y A L R 14 402 W K E L E M H S L A L Q N T L 14 407 M H S L A Q N T L H E R T W 14 424 E K N L M Q Q E L R S L K Q N 14 436 K Q N I F L F Y V K L R W L L 14 443 Y V K L R W L L K H W R Q G K 14 475 L S R L G E L G V Q G G H Q A 14 478 L G E L G V Q G G H Q A D G P 14 480 E L G V Q G G H Q A D G P D H 14 510 H S R V Q I G D H S L R L Q T 14 512 R V Q I G D H S L R L Q T A D 14 532 K Q V V E N Q Q L F S A F K A 14 545 K A L L E D F R A E L R E D E 14 562 R L R L Q Q Q Y A S D K A A W 14 575 A W D V E W A V L K C R L E Q 14 579 E W A V L K C R L E Q N C C G 14 594 Y P R I N I E E E T L G F T R 14 664 H S L V M D L R W Q I H H s E 14 667 V M D L R W Q I H H S E K N W 14 671 R W Q I H H S E K N w N R E K 14 712 Q G S L R M P R P V A M W P C 14 718 P R P V A M^' W P C A D A D S I 14 720 P V A M W P C A D A D S I P F 14 736 D R P L S K L K E S D R c S A 14 751 S E N L Y L D A L S ^■L D D E P 14 753 N L Y L D A L S L D D E P E E 14 756 L D A L S L D D Ξ P E E P P A 14 758 A L S L D D E P E E P P A H R 14 804 F Q R L M D I S P F L P E K G 14 807 L M D I S P F L P E K G L P S 14 816 E K G L P S T S S K E D V T P 14 825 K E D V T P P L S P D D L K Y 14 837 L K Y I E E F N K S W D Y T P 14 889 T T S V T M T T D T M T S P E 14 891 S V T M T T D T M T S P E H C 14 907 K Q P L R S H V L T E Q S G L 14 912 S H V L T E Q S G L R V L H S 14 930 V R R V D S I T A A G G Ξ G P 14 933 V D S I T A A G G E G P F P T 14 985 R R P L D S P L C T S L G F A 14 989 D S P L C T S L G F A S P L H 14 1002 L H S L E M S K N L S D D M K 14 1012 S D D M K E V A F S V R N A I 14 TABLE XXV - 185P2C9 v3 HLA- DRB1*0401 (DR4DW4) 15 - mers

Seq. ID

Pos • 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1019 A F S V R N A I C S G P G E L 14

1023 R N A I C S G P G E L Q V K D 14

1032 E L Q V K D M A C Q T N G S R 14

1035 V K D M A C Q T N G S R T M G 14

1045 S R T M G T Q T V Q T I S V G 14

1062 T E A L R G S G V T S s P H K 14

10^'86 A T P V S S P S R s L R s R Q 14

1102 A P A I E K V Q A K F E R T C 14

1167 H T T I N D G L S s L F N I I 14

1174 L S S L F N I I D H S P V V Q 14

1177 L F N I I D H s P V V Q D P F 14

1183 H S P V V Q D P F Q K G L R A 14

1207 R P E L G P G Q Ξ T G T N S R 14

1227 P I G V G S E M C R E E G G Ξ 14

1231 G S E M C R E E G G E G T P V 14

1246 K Q D L S A P P G Y T L T E N 14

1254 G Y T L T E N V A R I L N K K 14

1267 K K L L E H A L K E E R R Q A 14

TABLE XXVI - 185P2C9 v3 HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

315 L S T V T S V S R D S P I G N 28

324 D S P I G N L G K Ξ L G P D L 27

212 D S E Y L V T L K H E A Q R L 25

1175 S S L F N I I D H S P V V Q D 24

1188 Q D P F Q K G L R A G S R s R 24

437 Q N I F L F Y V K L R W L L K 22

686 V E L L D R L D R D R Q E W E 22

801 M S E F Q R L M D I S P F L P 22

1192 Q K G L R A G S R s R S A Ξ P 22

44 E E E A E L L R R s I S E I E 21

58 E D H N R Q L T H E L S K F K 21

711 E Q G S L R M P R P V A M W P 21

729 A D S I P F E D R P L S K L K 21

786 E N H K G N L Q R A V S V S S 21

1001 P L H s L E M S K N L S D D M 21

1008 S K N L S D D M K E V A F S V 21

1285 P P G L H S D S H S L G D T A 21

111 S G K V L K L Q H E H A L L 20

122 H A L L S N I Q R C D L A A H 20

213 S E Y L V T L K H E A Q R L E 20

288 L Q A F L Ξ Q V N R I G D G L 20

289 Q A F L E Q V N R I G D G L S 20

299 G D G L S P L P H L T E S S S 20

439 I F L F Y V K L R W L L K H w 20

443 Y V K L R w L L K H W R Q G K 20

463 G E E F ^■T E G E H P E T L S R 20

519 S L R L Q T A D R G Q P H K Q 20

683 R E K V E L L D R L D R D R Q 20

739 L S K L K E S D R C S A S E N 20

866 R T E V G R A G H E D S T E P 20

917 E Q S G L R V L H S P P A V R 20

924^' L H S P P A V R R V D S I T A 20

1099 R Q V A P A I E K V Q A K F E 20

1228 I G V G S E M C R E E G G E G 20.

1271 E H A L K Ξ E R R Q A A H G P 20 TABLE XXVI - 185P2C9 v3 HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

292 L E Q V N R I G D G L S P L P 19

311 S S S F L S T V T S V s R D s 19

395 E H L Y A L R W K E L E M H s 19

499 G C G F P V G E H S P H S R V 19

577 D V E W A V L K C R L E Q N C 19

804 F Q R L M D I S P F L P E K G 19

918 Q S G L R V L H s P P A V R R 19

977 S R D Y V E G A R R P L D S P 19

995 S L G F A S P L H S L E M S K 19

1174 L S S L F N I I D H S P V V Q 19

167 Q P K W Ξ G P V G G E S D S E 18

200 E P C P T Ξ L L K A R E D S E 18

237 T D S F L H D A G L R G G A P 18

302 L S P L P H L T E S S S F L s 18

475 L S R L G E L G V Q G G H Q A 18

548 L E D F R A E L R E D E R A R 18

603 T L G F T R L P A G S T V K T 18

696 R Q E W E R Q K K E F L W R I 18

846 S W D Y T P N R G H N G G G P 18

927 P P A V R R V D S I T A A G G 18

930 V R R V D S I T A A G G Ξ G P 18

1147 Q K G Y S E s A W A R S T T T 18

30 L E S S T E L R R H L Q F V E 17

68 L S K F K F E P P R E P G w L 17

272 Q L L G T I N A K M K A F K K 17

362 R L R A A R E L H R R A D G D 17

440 F L F Y V K L R W L L K H W R 17

445 K L R W L L K H W R Q G K Q M 17

941 G E G P F P T S R A R G S P G 17

942 E G P F P T s R A R G S P G D 17

966 L S R W P c T S P R H s R D Y 17

1017 E V A F s V R N A I C s G P G 17

78 E P G W L G E G A S P G A G G 16

181 E E M F E K T S G F G s G K P 16

187 T S G F G S G K P S E A S E P 16

354 R G D E R E S L R L R A A R E 16

399 A L R W K E L E M H S L A L Q 16

409 S L A L Q N T L H E R T W S D 16

411 A L Q N T L H E R T W S D E K 16

444 V K L R w L L K H W R Q G K Q 16

537 N Q Q L F S A F K A L L E D F 16

541 F S A F K A L L E D F R A E L 16

552 R A E L R E D E R A R L R L Q 16

576 w D V E W A V L K C R L E Q N 16

591 C C G Y P R I N I E E E T L G 16

643 A E S K G A L K K E R E V H Q 16

650 K K E R E V H Q K L L A D S H 16

703 K K E F L W R I E Q G S L R M 16

809 D I S P F L P E K G L P S T S 16

810 I S P F L P E K G L P s T s S 16

840 I E E F N K S W D Y T P N R G 16

863 w A D R T J3 V G R A G H E D S 16

896 T D T M T S P E H C Q K Q P L 16

943 G P F P T S R A R G S P G D T 16

1015 M K E V A F S V R N A I C s G 16

1028 S G P G E L Q V K D M A C Q T 16

1038 M A C Q T N G S R T M G T Q T 16

1058 V G L Q T E A L R G S G V T s 16 cn cn cn ιπ ιπ Lπ rf* rf* ιf* rf* ιo ιo Lo ιo to to to tθ H H H H H H _lr, ~. H H H H H H H H H iD vi Ui Oi ui ^ iMiJ ω i w u i μ - . ^

H H o rf* ιf* H vo σι ιf* ω cD cn rf* w _^_ _o _o o _^] cn rf* ιo _o o ,j 5_ vo --α tO tO tO tO tO H H O O en to vo oo ^ W W H H U O -. („ O ^ U W H ^ " " ° O a

~j ιπ o rf_* to -j o vD cn oo cD rf_* ιf_* ιπ rf* vo rf* to co H ι to o oo o cn in ui io o rf* vo tfi vo rf* H H cn oo ιo cn H θ Lπ o to ιo ιπ _o rf_* o rf_* ^{u, l, 1,}^ ^lw r_. ^u} co

rt < B a cn O o Ω a a a a B Ω a Ω to a α a cn Ω a a KD a a B Ω to a a a to a a a a a a a KD a ω a <: Ω Ω a a a a to a Ω a a H o a Ω a rt a σ a α a Ω B to rt a KD α a a a rt Ω a Ω a rt o a KD Ω a a a a to a a a a D cn a o cn a rt a to a to a a 2 a a to a o a α rt cn cn to D a a a a B rt a a a a cn a B a a a 2 D to a a a a a a a a a a Ω o to a rt a a a σ a a a B a B a a rt a B a B to a co a B Ω a a to a to rt Ω a a a < <! α a rt < 3 a Ω > a 3 B > a a rt a rt < rf_* a to a a a a a a a a a 2 a 2 XD a a a a a a a ot Ω D a a a Ω a a a a rt a cn a a a KD KD a a a rt a a KD D a a B te Ω hi H

*

Ω cn Ω a a a o rt a B a KD to to to cn a 2 a Θ a a B Ω Ω o a a a a rt a a cn Ω a a H a rt B a to rt a a B B α B rt cn en H H a a a o a KD cn a a < a a a a a Ω a a a KD > Ω X < B a a a M a a < <! H O a a a α H Ω a B Ω a O a <! a B a a X a a o o

C7\ H O KD Ω rt a a tr α to 2 a B a Ω a 3 a a to B a Ω a a σ Ω B B a a to a a a cn co a cn a α a 3 co a n a a B a a a a cn a <_n H a a a a α a a a a a a a a a a a a a a a a Ω a a a a a a a a a a to α a a a a a X a a a a a a to a a a a a a a a a a vo in a a to a α Ω a KD a a to to B to Ω o a rt 2 a to a a co a to o B a to a o Ω a KD a a en en Ω a a B a H > a a a Ω a o a a a B a a Ω Ω Ω 2 o a a Ω > o B a a a KD B a a Ω KD a a a a a a < a a a a a a tri tr tr te a a < a a a a a Ω 3 a a to a to Ω Ω B a a to Ω Ω a a to Ω cn Ω Ω to to a a cn to a te a a a co cn a to a α B <; Ω a a to Ω o CO KD rt a a Ω a cn a a a Ω a B D a a D a a α o a a a a B •fl a o a a a a Ω to a a a > to a a co 2 a a KD a Ω Ω rt to O a cn 2 B X a rt a a a a a a a Ω 2 a a rt B 2 a a a Ω Ω a co a KD a a cn a a a a a a a to > a a σ OOO a o KD α a rt a a a a B a a rf_* a a rt α a a < <! a a 2 Ω a KD a Ω a cn Ω a D a a H a rt α Ω a to a a a α KD a a a a rt a rt rt rt a D H H Ω a a a

H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H n it* rf* it* rf_* rf* rf_* rf_* rf_* rf_* rf_* rf* rf* rf_* rf_* rf_* rf_* rf* rf* rf_* rf_* if* rf* rf* it* rf_* rf_* rf_* tfl in in tfl in tfl in UI in LΠ tfl tfl in tfl tfl in in tfl in tfl tfl tfl tfl ui tfl tfl in cn n

ro

a H

TABLE XXVI - 185P2C9 v3 HLA- DRB1*1101 15 - mers

Seq . ID

Pos . 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

623 L Q R L E L E E K T E N K L G 14

663 S H S L V M D L R W Q I H H S 14

667 V M D L R W Q I H H S E K N W 14

668 M D L R W Q I H H S E K N w N 14

671 R W Q I H H S E K N W N R E K 14

701 R Q K K E F L W R I E Q G S L 14

709 R I E Q G S L R M P R P V A M 14

714 S L R M P R P V A M W P C A D 14

789 K G N L Q R A V S V s s M S E 14

798 V S S M S E F Q R L M D I S P 14

837 L K Y I E E F N K S W D Y T P 14

847 W D Y T P N R G H N G G G P D 14

891 S V T M T T D T M T S P E H C 14

933 V D S I T A A G G E G P F P T 14

950 A R G S P G D T K G G P P E P 14

960 G P P E P M L S R W P C T S P 14

990 S P L C T S L G F A S P L H s 14

1073 S P H K c L T P K A G G G A T 14

1075 H K C L T P K A G G G A T P V 14

1112 F E R T c C S P K Y G S P K L 14

1120 K Y G S P K L Q R K P L P K A 14

1125 K L Q R K P L P K A D Q P N N 14

1132 P K A D Q P N N R T S P G M A 14

1140 R T S P G M A Q K G Y s E S A 14

1154 A w A R S T T T R E S P V H T 14

1215 E T G T N S R G R S P s P I G 14

1267 K K L L E H A L K E E R R Q A 14

1275 K E E R R Q A A H G P P G L H 14

1281 A A H G P P G L H S D S H S L 14

97 Q E E L K S A R L Q I S E L S 13

104 R L Q I S E L S G K V L K L Q 13

127 N I Q R C D L A A H L G L R A 13

131 C D L A A H L G L R A P S P R 13

136 H L G L R A P S P R D S D A E 13

180 S E E M F E K T S G F G S G K 13

238 D S F L H D A G L R G G A P L 13

295 V N R I G D G L S P L P H L T 13

340 S R L K E Q L E-W Q L G P A R 13

355 G D E R E S L R L R A A R Ξ L 13

429 Q Q E L R S L K Q N I F L F Y 13

478 L G E L G V Q G G H Q A D G P 13

498 R G C G F P V G E H S P H S R 13

594 Y P R I N I Ξ E E T L G F T R 13

609 L P A G S T V K T L K S L G L 13

636 L G E L G S S A E S K G A L K 13

664 H S L V M D L R W Q I H H S E 13

707 L W R I E Q G S L R M P R P V 13

807 L M D I S P F L P E K G L P S 13

822 T S S K E D V T P P L S P D D 13

859 G P D L W A D R T E V G R A G 13

909 P L R S H V L T E Q S G L R V 13

959 G G P P E P M L S R w P C T S 13

986 R P L D S P L C T S L G F A S 13

999 A S P L H s L E M S K N L S D 13

1012 S D D M K E V A F S V R N A I 13

1050 T Q T V Q T I S V G L Q T E A 13

1053 V Q T I S V G Q T E A L R G 13

1057 S V G L Q T E A L R G S G V T 13 TABLE XXVI - 185P2C9 v3 HLA- DRB1* 1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

1123 K L Q L P K A D P 13 1171 G L S N I I D H P 13 1183 P V V F Q K G L A 13 1251 P G Y E N V A R L 13 1258 N V A N K K L L E H 13 http://ww .b_m-heidelberg.com/syfpeithi

TABLE XXIII - 185P3C2 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

109 P G S L P P L D S Ξ D L F Q D 30

8 N K S V R W G P G A G A A L L 28

74 A G Y L D Q Q V P Y T F S S K • 28

100 I G P L G K L M D P G S L P P 28

10 s V R W G P G A G A A L L R P 26

259 c H S F T s Q G G G R E P L P 26

96 R Ξ A L I G P L G K L M D P G 25

227 R N F L R S s G T S Q P H P G 25

253 Q Q P L D I c H S F T S Q G G 25

66 G R M E R R M K A G Y L D Q Q 24

241 G H G Y L G E H S S V F Q Q P 24

16 G A G A A L L R P S P A A L G 23

78 D Q Q V P Y T F S S K S P G N 23

103 L G K L M D P G S L P P L D S 23

196 A Y D P P R Q I A I K S P A P 23

202 Q I A I K S P A P G A L G Q S 23

18 G A A L L R P S P A A L G A G 22

44 A Q T P R P Q V s A P A W G P 22

101 G P L G K L M D P G S L P P L 22

106 L M D P G S L P P L D S E D L 22

122 Q D L S H F Q E T W L A E A Q 22

142 E Q F V P D F H s E N L A F H 22

247 E H S S V F Q Q P L D I c H S 22

273 P A P Y Q H Q L s E P c P P Y 22

277 Q H Q L S E P C P P Y P Q Q S 22

32 G S R A C S V P P A A P A Q T 20

83 Y T F S s K s P G N G s L R E 20

199 P P R Q I A I K s P A P G A L 20

201 R Q I A I K s P A P G A L G Q 20

209 A P G A L G Q s P L Q P F P R 20

218 L Q P F P R A E Q R N F L R S 20

47 P R P Q V S A P A W G P G R A 19

54 P A W G P G R A A R G S G R M 19

226 Q ^•R N F L R S S G T S Q P H P 19

6 P K N K s V R w G P G A G A A 18

19 A A L L R P S P A A L G A G S 18

20 A L L R P S P A A L G A G S R 18

22 L R P S P A A L G A G S R A C 18

53 A P A W G P G R A A R G S G R 18

92 N G S L R E A L I G P L G K L 18

118 E D L F Q D L S H F Q E T W L 18

124 L S H F Q E T W L A E A Q V P 18

127 F Q E T W L A E A Q V P D s D 18

194 S S A Y D P P R Q I A I K s P 18

224 A E Q R N F L R S s G T S Q ^'p 18

260 H S F T S Q G G G R E P L P A 18 TABLE XXIII - 185P3C2 HLA- DRB1*0101 15 - mers

Seq . ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num .

9 K S V R W G P G A G A A L L R 17

11 V R W G P G A G A A L L R P S 17

23 R P S P A A L G A G S R A C S 17

24 P S P A A L G A G S R A C S V 17

51 V s A P A W G P G R A A R G S 17

97 E A L I G P L G K L M D P G S 17

102 P L G K L M D P G S L P P L D 17

121 F Q D L S H F Q E T W L A E A 17

126 H F Q E T W L A E A Q V P D S 17

152 N L A F H S P T T R I K K E P 17

158 P T T R I K K E P Q s P R T D 17

170 R T D P A L S C S R K P P L P 17

197 Y D P P R Q I A I K s P A P G 17

200 P R Q I A I K S P A P G A L G 17

207 S P A P G A L G Q S P L Q P F 17

239 H P G H G Y L G E H s S V F Q 17

284 C P P Y P Q Q S F K Q E Y H D 17

15 P G A G A A L L R P s P A A L 16

26 P A A L G A G S R A c S V P P 16

27 A A L G A G S R A C s V P P A 16

33 S R A C S V P P A A P A Q T P 16

36 C S V P P A A P A Q T P R P Q 16

71 R M K A G Y L D Q Q V P Y T F 16

73 K A G Y L D Q Q V P Y T F S S 16

82 P Y T F S S K S P G N G S L R 16

89 S P G N G S L R E A L I G P L 16

93 G S L R E A L I G P L G K L M 16

94 S L R E A L I G P L G K L M D 16

104 G K L M D P G S L P P L D S E 16

114 P L D S E D L F Q D L S H F Q 16

129 E T W L A E A Q V P D S D E Q 16

134 E A Q V P D S D E Q F V P D F 16

146 P D F H S E N L A F H S P T T 16

147 D F H S E N L A F H S P T T R 16

150 S E N L A F H S P T T R I K K 16

172 D P A L S C S R K P P L P Y H 16

180 K P P L P Y H H G E Q C L Y S 16

188 G E Q C L Y S S A Y D P P R Q 16

189 E Q C L Y s S A Y D P P R Q I 16

206 K S P A P G A L G Q s P L Q P 16

210 P G A L G Q S P L Q P F P R A 16

238 P H P G H G Y L G E H S S V F 16

250 S V F Q Q P L D I C H S F T S 16

266 G G G R E P L P A P Y Q H Q L 16

269 R E P L P A P Y Q H Q L S E P 16

274 A P Y Q H Q L S E P C P P Y P 16

275 P Y Q H Q L S E P C P P Y P Q 16

17 A G A A L L R P S P A A L G A 15

50 Q V S A P A W G P G R A A R G 15

57 G P G R A A R G S G R M E R R 15

75 G Y L D Q Q V P Y T F S S K S 15

79 Q Q V P Y T F S s K S P G N G 15

116 D S E D L F Q D L s H F Q E T 15

139 D s D E Q F V P D F H S E N L 15

151 E N L A F H S P T T R I K K E 15

192 L Y S S A Y D P P R Q I A I K 15

213 L G Q S P L Q P F P R A E Q R 15,

225 E Q R N F L R S S G T S Q P H 15 TABLE XXIII - 185P3C2 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

263 T S Q G G G R E P L P A P Y Q 15 29 L G A G S R A C s V P P A A P 14 31 A G S R A C S V P P A A P A Q 14 35 A C S V P P A A P A Q T P R P 14 38 V P P A A P A Q T P R P Q V s 14 41 A A P A Q T P R P Q V S A P A 14 45 Q T P R P Q V S A P A W G P G 14 76 Y L D Q Q V P Y T F S s K S P 14 117 S E D L F Q D L S H F Q E T W 14 137 V P D S D E Q F V P D F H S E 14 163 K K E P Q S P R T D P A L s C 14 165 E P Q S P R T D P A L s C s R 14 173 P A L S C S R K P P L P Y H H 14 184 P Y H H G E Q C L Y S s A Y D 14 186 H H G E Q C L Y S s A Y D P P 14 212 A L G Q S P L Q P F P R A E Q 14 242 H G Y L G E H S s V F Q Q P L 14 245 L G E H S S V F Q Q P L D I C 14 248 H S S V F Q Q P L D I C H s F 14 255 P L D I C H S F T S Q G G G R 14 280 L S E P C P P Y P Q Q S F K Q 14

TABLE XXIV - 185P3C2 HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

110 G S L P P L D_. S E D L F Q D L 27

72 M K A G Y L D Q Q V P Y T F S 26

65 S G R M E R R M K A G Y L D Q 25

121 F Q D L S H F Q E T W L A E A 24

172 D P A L S C S R K P P L P Y H 24

218 L Q P F P R A E Q R N F L R S 24

269 R E P L P A P Y Q H Q L S E P 24

18 G A A L L R P S P A A L σ A G 23

92 N G S L R E A L I G P L G K L 21

113 P P L D S Ξ D L F Q D L s H F 21

117 S E D L F Q D L S H F Q E T W 21

133 A E A Q V P D S D E Q F V P D 21

202 Q I A I K S P A P G A L G Q S 21

104 G K L M D P G S L P P L D S Ξ 20

141 D E Q F V P D F H S E N L A F 20

210 P G A L G Q S P L Q P F P R A 20

97 E A L I G P L G K L M D P G S^' 19

109 P G S L P P L D S E D L F Q D 19

142 E Q F V P D F H S E N L A F H 19

215 Q S P L Q P F P R A E Q R N F 19

251 V F Q Q P L D I C H S F T S Q 19

1 N C L L R P K N K S V R W G P 18

26 P A A L G A G S R A c S V P P 18 74 A G Y L D Q Q V P Y T F s S K 18 135 A Q V P D S D E Q F V P D F H 18 159 T T R I K K E P Q S P R T D P 18 84 T F S s K S P G N G s L R E A 17 134 E A Q V P D S D E Q F V P D F 17 144 F V P D F H S E N L A F H S P 17 281 S E P C P P Y P Q Q S F K Q E 17 88 K S P G N G S L R E A L I G P 16 181 P P L P Y H H G E Q c L Y S S 16 TABLE XXIV - 185P3C2 HLA- DRB1* 0301 (DR17 ) 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 Seq. ID

3 4 5 score Num.

182 P L P Y H H G E Q c L Y S S A 16 219 Q P F P R A E Q R N F L R S S 16 245 L G E H S S V F Q Q P L D I C 16

19 A A L L R P S P A A L G A G S 15

102 P L G K L M D P G S L P P L D 15

207 S P A P G A L G Q S P L Q P F 15

261 S F T S Q G G G R E P L P A P 15

8 N K S V R W G P G A G A A L L 13

96 R E A L I G P L G K L M D P G 13 166 P Q S P R T D P A L S C S R K 13 227 R N F L R S S G T S Q P H P G 13 242 H G Y L G E H S S V F Q Q P L 13

Table XXV - 185P3C2 HLA- DRB1*0401 (DR4D 4 ) 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

141 D E Q F V P D F H S E N L A F 28

109 P G S L P P L D s E D L F Q D 26

117 S E D L F Q D L s H F Q E T W 26

142 E Q F V P D F H s E N L A F H 26

215 Q S P L Q P F P R A Ξ Q R N F 26

118 E D L F Q D L S H F Q E T W L 22

128 Q E T W L A E A Q V P D S D E 22

152 L A F H S P T T R I K K E P 22

194 S S A Y D P P R Q I A I K S P 22

218 L Q P F P R A E Q R N F L R S 22

241 G H G Y L G E H S S V F Q Q P 22

26 P A A L G A G S R A C S V P P 20

35 A C S V P P A A P A Q T P R P 20

78 D Q Q V P Y T F s S K s P G N 20

92 N G S L R E A L I G P L G K L 20

100 I G P L G K L M D P G s L P P 20

121 F Q D L S H F Q E T W L A E A 20

129 E T W L A E A Q V P D S D E Q 20

159 T T R I K K E P Q S P R T D P 20

242 H G Y L G E H s S V F Q Q P L 20

253 Q Q P L D I C H S F T S Q G G 20

269 R E P L P A P Y Q H Q L S E P 20

63 R G S G R M E R R M K A G Y L 18

114 P L D S E D L F Q D L S H F Q 18

148 F H S E N L A F H S P T T R I 18

149 H S E N L A F H S P T T R I K 18

207 S P A P G A L G Q S P L Q P F 18

219 Q P F P R A E Q R N F L R s S 18

224 A E Q R N F L R S S G T S Q P 18

245 L G E H S S V F Q Q P L D I c 18

252 F Q Q P L D I C H S F T S Q G 18

10 S V R W G P G A G A A L L R P 16

73 K A G Y L D Q Q V P Y T F S s 16

124 L S H F Q E T W L A E A Q V P 16

145 V P D F H S E N L A F H S P T 16

182 P L P Y H H G E Q C L Y S s A 16

190 Q C L Y S S A Y D P P R Q I A 16

226 Q R N F L R S S G T S Q P H P 16

273 P A P Y Q H Q L S E P C P P Y 16

8 N K S V R W G P G A G A A L L 14

18 G A A L L R P s P A A L G A G 14 Table XXV - 185P3C2 HLA- DRB1*0401 (DR4Dw4 ) 15 - mers

Pos 1 2 3 4 5 6 7 8 9 0 1 2 Seq. ID

3 4 5 score Num.

69 E R R M K A G Y L D Q Q V P Y 14 96 R E A L I G P L G K L M D P G 14

97 E A L I G P L G K L M D P G S 14 103 L G K L M D P G S L P P L D S 14 104 G K L M D P G S L P P L D S E 14 112 L P P L D S E D L F Q D L S H 14 134 E A Q V P D S D E Q F V P D F 14 172 D P A L S C S R K P P L P Y H 14 202 Q I A I K S P A P G A L G Q S 14 227 R N F L R S S G T S Q P H P G 14 255 P L D I C H S F T S Q G G G R 14

TABLE XXVI - 185P3C2 HLA- DRB1*1101 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

80 Q V P Y T F S S K s P G N G S 26

118 E D L F Q D L S H F Q E T W L 24

15 P G A G A A L L R P S P A A L 20

26 P A A L G A G S R A C S V P P 20

97 E A L I G P L G K L M D P G S 20

53 A P A W G P G R A A R G S G R 18

100 I G P L G K L M D P G S L P P 18

141 D E Q F V P D F H s E N L A F 18

4 L R P K N K S V R w G P G A G 16

82 P Y T F S S K S P G N G S L R 16

124 L S H F Q E T W L A E A Q V P 16

155 F H S P T T R I K K Ξ P Q S P 16

156 H S P T T R I K K E P Q S P R 16

1 N C L L R P K N K S V R W G P 15

62 A R G S G R M E R R M K A G Y 15

63 R G S G R M E R R M K A G Y L 15

193 Y S S A Y D P P R Q I A I K S 15

270 E P L P A P Y Q H Q L S E P C 15

55 A W G P G R A A R G S G R M E 14

65 S G R M E R R M K A G Y L D Q 14

162 I K K Ξ P Q S P R T D P A L S 14

171 T D P A L S C S R K P P L P Y 14

172 D P A L S C S R K P P L P Y H 14

198 D P P R Q I A I K s P A P G A 14

215 Q S P L Q P F P R A E Q R N F 14

223 R A E Q R N F L R S S G T S Q 14

252 F Q Q P L D I C H s F T S Q G 14

259 C H S F T S Q G G G R E P L P 14

19 A A L L R P S P A A L G A G s 13

71 R M K A G Y L D Q Q V P Y T F 13

78 D Q Q V P Y T F S s K S P G N 13

93 G S L R E A L I G P L G K L M 13

114 P L D S E D L F Q D L S H F Q 13

5 R P K N K S V R W G P G A G A 12

8 N K S V R W G P G A G A A L L 12

16 G A G A A L L R P S P A A L G 12 32 G S R A C S V P P A A P A Q T 12 48 R P Q V S A P A w G P G R A A 12 75 G Y L D Q Q V P Y T F S S K s 12 103 L G K L M D P G s L P P L D S 12 106 L M D P G S L P P L D S E D L 12 109 P G S L P P L D s E D L F Q D 12 TABLE XXVI - 185P3C2 HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

129 E T W L A E A Q V P D S D E Q 12 139 D S D E Q F V P D F H S E N L 12 147 D F H S E N L A F H S P T T R 12 152 N L A F H S P T T R I K K E P 12 177 C S R K P P L P Y H H G E Q C 12 189 E Q C L Y S S A Y D P P R Q I 12 197 Y D P P R Q I A I K S P A P G 12 199 P P R Q I A I K s P A P G A L 12 200 P R Q I A I K S P A P G A L G 12 227 R N F L R s s G T S Q P H P G 12 239 H P G H G Y L G E H s S V F Q 12 250 S V F Q Q P L D I C H S F T S 12 253 Q Q P L D I C H s F T S Q G G 12 255 P L D I C H S F T S Q G G G R 12 274 A P - Y Q H Q L S E P C P P Y P 12 277 Q H Q L S E P C P P Y P Q Q S ^" 12

TABLE XXIII - 186P1H9 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

80 A R A I R E L T G K L A R C E 35 43 L P A M P M Q G G A Q S P Ξ E 32 240 K K T L P E L Y A F T I C L W 32 119 P G H V V E Q L S R S L Q T L 30 215 N S A F K S P D A F K V S L P 28 6 A A S V A L A V A A G A Q D S 27

93 C E G L A G G K A R G A G A T 27

233 N Y L Y G K I K K T L P E L Y 27

276 N E I L L I E w G N N P I E L 27

154 P G D F R E V L Q Q R L G E L 26

173 L R K V A E L E D E K S L L H 26

205 L Q R V T Ξ L E R G N s A F K 26

252 C L W L R S S A S P G I G T P 26

60 R A A V L Q L R E T V V Q Q K 25

90 L A R C E G L A G G K A R G A 25

162 Q Q R L G E L E R Q L L R K V 25

2 L A L L A A S V A L A V A A G 24

37 V H A G C P L P A M P M Q G G 24

53 Q S P E E E L R A A V L Q L R 24

67 R E T V V Q Q K E T L A S A R 24

112 M G D L P R D P G H V V E Q L 24

138 E S L E H Q L R A N V s N A G 24

165 L G E L E R Q L L R K V A Ξ L 24

264 G T P F S Y A V P G Q A N E I 24

22 A P G S R F V C T A L P P E A 23

26 R ^'F V C T A L P P E A V H A G 23

68 E T V V Q Q K E T L A S A R A 23

250 T I C L W L R S S A S P G I G 23

25 S R F V C T A L P P E A V H A 22

31 A L P P E A V H A G C P L P A 22

34 P E A V H A G C P L P A M P M 22

71 V Q Q K E T L A S A R A I R E 22 ^•

137 L E s L E H Q L R A N V s N A 22

157 F R E V L Q Q R L G E L E R Q 22

224 F K V S L P L R T N Y L Y G K 22

246 L Y A F T I C L w L R S s A .S 22

249 F T I C L w L R s S A S P G I 22 TABLE XXIII - 186P1H9 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

260 S P G I G T P F S Y A V P G Q 22

268 S Y A V P G Q A N E I L L I E 22

257 S S A s P G I G T P F S Y A V 21

12 A V A A G A Q D S P A P G S R 20

95 G L A G G K A R G A G A T G K 20

72 Q Q K E T L A S A R A I R E L 19

166 G E L E R Q L L R K V A E L E 19

209 T E L E R G N S A F K S P D A 19

244 P E L Y A F T I C L W L R S S 19

9 V A L A V A A G A Q D S P A P 18

14 A G A Q D S P A P G s R F V 18 83 I R E L T G K L A R C E G L A 18 170 R Q L L R K V A E L E D E K S 18 198 E S T L N A L L Q R V T Ξ L E 18 229 P L R T N Y L Y G K I K K T L 18 251 I C L W L R S S A S P G I G T 18 280 L I E W G N N P I E L L I N D 18 28 V C T A L P P E A V H A G C P 17 44 P A M P M Q G G A Q S P Ξ E E 17 87 T G K L A R C E G L A G G K A 17 105 G T G K D T M G D L P R D P 17 124 E Q L S R S L Q T L K D R L E 17 127 S R S L Q T L K D R L E S L E 17 134 K D R L E s L E H Q L R A N V 17 143 Q L R A N V S N A G L P G D F 17 180 E D E K S L L H N E T S A H R 17 206 Q R V T E L Ξ R G N S A F K S 17 221 P D A F K V S L P L R T N Y L 17 232 T N Y L Y G K I K K T L P E L 17 236 Y G K I K K T L P E L Y A F T 17 237 G K I K K T L P E L Y A F T I 17 247 Y A F T I c L W L R S S A S P 17 248 A F T I c L W L R S S A S P G 17 256 R S S A s P G I G T P F S Y A 17 267 F S Y A V P G Q A N E I L L I 17 277 E I L L I E W G N N P I E L L 17

TABLE XXIV - 186P1H9 HLA- DRB1*0301 (DR17 ) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

150 N A G L P G D F R E V I" Q Q R 34

175 K V A E L E D E K S L L H N E 29

162 Q Q R L G E L E R Q L L R K V 28

223 A F K V S L P L R T N Y L Y G 27

154 P G D F R E V L Q Q R L G E L 26

225 K V S L P L R T N Y L Y G K I 26

232 T N Y L Y G K I K K T L P E L 24

268 s Y A V P G Q A N E I L L I E 24

112 M G D L P R D P G H V V E Q L 21

119 P G H V V E Q L S R s I. Q T L 21

236 Y G K I K K T L P E L Y A F T 21

129 S L Q T L K D R L E s L.E H Q 20

157 F R E V L Q Q R L G E L E R Q 20

205 ^' L Q R V T E L E R G N S A F K 20

277 E I L L I E W G N N P I E L L 20

67 R E T V V Q Q K E T L A S A R 19-

130 L Q T L K D R L E S L E H Q L 19 TABLE XXIV - 186P1H9 HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

137 L E S L E H Q L R A N V S N A 19 207 R V T E L E R G N S A F K S P 19 60 R A A V L Q L R E T V V Q Q K 18 104 A G A T G K D T M G D L P R D 18 120 G H V V E Q L S R S L Q T L K 18 123 V E Q L S R S L Q T L K D R L 18 133 L K D R L E S L E H Q L R A N 18 170 R Q L L R K V A E L E D E K S 18 194 R Q K T E S T L N A L L Q R V 18 248 A F T I C L w L R S S A S P G 18 109 K D T M G D L P R D P G H V V 17 126 L S R S L Q T L K D R L E S L 17 161 L Q Q R L G E L E R Q L L R K 17 198 E S T L N A L L Q R V T E L E 17 202 N A L L Q R V T E L E R G N S 17 10 A L A V A A G A Q D S P A P G 16 45 A M P M Q G G A Q S P E Ξ E L 16 142 H Q L R A N V S N A G L P G D 16 278 I L L I E W G N N P I E L L I 16

TABLE XXV - 186P1H9 HLA- DRB1*0401 (DR4DW4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

154 P G D F R E V L Q Q R L G Ξ L 28

120 G H V V E Q L S R s L Q T L K 26

150 N A G L P G D F R E V L Q Q R 26

162 Q Q R L G Ξ L E R Q L L R K V 26

198 E S T L N A L L Q R V T E L E 26

205 L Q R V T E L E R G N S A F K 26

264 G T P F S Y A V P G Q A N E I 22

1 M L A L L A A S V A L A V A A 20

6 A A S V A L A V A A G A Q D S 20

25 S R F V C T A L P P Ξ A V H A 20

60 R A A V L Q L R E T V V Q Q K 20

80 A R A I R E L T G K L A R C Ξ 20

109 K D T M G D L P R D P G H V V 20

112 M G D L P R D P G H V V E Q L 20

119 P G H V V E Q L S R S L Q T L 20

123 V E Q L S R S L Q T L K D R L 20

134 K D R L E S L E H Q L R A N V 20

137 L E S L E H Q L R A N V S N A 20

173 L R K V A E L E D E K S L L H 20

176 V A E L E D E K S L L H N E T 20

183 K S L L H N E T S A H R Q K T 20

208 V T E L E R G N S A F K S P D 20

240 K K T L P E L Y A F T I C L W 20

248 A F T I C L W L R S S A s P G 20

252 C L W L R S S A S P G I G T P 20

260 S P G I G T P F S Y A V P G Q 20

268 S Y A V P G Q A N E I L L I E 20

275 A N E I L L I E W G N N P I E 20

277 E I L L I E w G N N P I E L L 20

52 A Q S P E E E L R A A V L Q L 18

65 Q L R E T V V Q Q K E T L A S 18

71 V Q Q K E T L A S A R A I R E 18

76 T L A S A R A I R Ξ L T G K L 18

79 S A R A I R E L T G K L A R C 18 TABLE XXV - 186P1H9 HLA- DRB1*0401 (DR4Dw4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

135 D R L E S L E H Q L R A N V S 18 142 H Q L R A N V S N A G L P G D 18 166 G E L E R Q L L R K V A E L E 18 179 L E D E K S L L H N E T S A H 18 180 E D E K S L L H N E T S A H R 18 186 L H N E T S A H R Q K T E S T 18 192 A H R Q K T E S T L N A L L Q 18 212 E R G N S A F K s P D A F K V 18 224 F K V S L P L R T N Y L Y G K 18 242 T L P Ξ L Y A F T I C L W L R 18 249 F T I C L W L R s s A S P G I 18 257 S S A S P G I G T P F S Y A V 18 215 N S A F K S P D A F K V S L P 16 221 P D A F K V S L P L R T N Y L 16 231 R T N Y L Y G K I K K T L P E 16 233 N Y L Y G K I K K T L P E L Y 16 244 P E L Y A F T I c L W L R S S 16 246 L Y A F T I c L W L R S S A S 16 266 P F S Y A V P G Q A N E I L L 16 280 L I E W G N N P I Ξ L L I N D 16 61 A V L Q L R Ξ T V V Q Q K E 15 68 E T V V Q Q K Ξ T L A S A R A 15 2 L A L L A A S V A L A V A A G 14 8 S V A L A V A A G A Q D S P A 14

10 A L A V A A G A Q D S P A P G 14 29 C T A L P P E A V H A G C P L 14 34 P E A V H A G C P L P A M P M 14 43 L P A M P M Q G G A Q S P E E 14 45 A P M Q G G A Q S P E E E L 14 56 E E E L R A A V L Q L R E T V 14 63 V L Q L R E T V V Q Q K Ξ T L 14 67 R E T V V Q Q K E T L A S A R 14 74 K E T L A S A R A I R E L T G 14 93 C E G L A G G K A R G A G A T 14 127 S R S L Q T L K D R L E S L E 14 141 E H Q L R A N V s N A G L P G 14 145 R A N V S N A G L P σ D F R E 14 157 F R E V L 'Q Q R L G Ξ L E R Q 14 165 L G E L E R Q L L R K V A Ξ L 14 170 R Q L L R K V A E L E D E K S 14 182 E K S L L H N E T S A H R Q K 14 202 N A L L Q R V T Ξ L E R G N S 14 223 A F K V S L P L R T N Y L Y G 14 227 S L P L R T N Y L Y G K I K K 14 243 L P E L Y A F T I c L W L R S 14 276 E I L L I E W G N N P I E L 14 278 I L I E w G N N P I E L L I 14

TABLE XXVI - 186P1H9 HLA- DRB1*1101 15 - mers

Seq. ID os 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num. 33 N Y L Y G K I K K T L P E L Y 31 05 L Q R V T E L E R G N S A F K 28 09 K D T M G D L P R D P G H V V 26 20 G H V V E Q L S R S L Q T L K 21 34 K D R L E S L Ξ H Q L R A N V 21 62 Q Q R L G E L E R Q L L R K V 21 TABLE XXVI - 186P1H9 HLA- DRB1*1101 15 - mers q. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 Se score Num.

166 G E L E R Q L L R K V A E L Ξ 21

84 R Ξ L T G K L A R c E G L A G 20 246 L Y A F T I C L W L R S S A S 20 248 A F T I C L W L R S S A S P G 20

80 A R A I R E L T G K L A R c E 19 173 L R K V A E L E D E K S L L H 19

40 G C P L P A M P M Q G G A Q S 18 154 P G D F R E V L Q Q R L G E L 18 264 G T P F S Y A V P G Q A N E I 18

92 R C E G L A G G K A R G A G A 16 128 R S L Q T L K D R L E S L E H 16 251 I C L W L R S S A S P G I G T 16 116 P R D P G H V V E Q L S R S L 15 150 N A G L P G D F R E V L Q Q R 15 155 G D F R E V L Q Q R L G E L E 15 httρ://ww .br__i-heidelberg.coιι_/syr eithi

TABLE XXIII - 187P3F2 (500a. a.) HLA-DRB1*0101 15 mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

67 P S S V K M V Q S D F M Q G A 30

227 G F T V N G M L S A P P G P G 29

9 Y L P G N S L L A A G S I V H 27

18 A G s I V H S D A A G A G G G 27

219 S L L Y S Q P G G F T V N G M 27

184 P G G w G A A A A A A A A A A 26

10 L P G N S L L A A G S I V H S 25

94 A H Q W V T A L P H A A A A A 25

95 H Q W V T A L P H A A A A A A 25

^■4 A A s N P Y L P G N S L L A A 24

6 S N P Y L P G N S L L A A G S 24

73 V Q s D F M Q G A M A A S N G 24

79 Q G A M A A S N G G H M L S H 24

98 V T A L P H A A A A A A A A A 24

152 G A G R D D L H A G T A L H H 24

217 P Q S L L Y S Q P G G F T V N 24

257 P G L V R G D T P E L A E H H 24

16 L A A G S I V H S D A A G A G 23

46 G G G G G M Q P G S A A V T S 23

72 M V Q S D F M Q G A M A A S N 23

110 A A A A A A V E A S S P W S G 23

161 G T A L H H R G P P H L G P P 23

166 H R G P P H L G P P P P P P H 23

254 L V H P G L V R G D T P E L A 23

256 H P G L V R G D T P E L A E H 23

203 A A H L P S M A G G Q Q P P P 22

218 Q S L L Y S Q P G G F T V N G 22

224 Q P G G F T V N G M L S A P P 22

299 G G A G P G L N S H D P H S D 22

249 G G A Q S L V H P G L V R G D 21

45 G G G G σ G M Q P G S A A V T 20

128 G M A G s P Q Q P P Q P P P P 20

129 M A G S P Q Q P P Q P P P P P 20

132 S P Q Q P P Q P P P P P P Q G 20 TABLE XXIII - 187P3F2 (500a. a.) HLA-DRB1*0101 15 mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

158 L H A G T A L H H R G P P H L 20

208 S M A G G Q Q P P P Q S L L Y 20

225 P G G F T V N G M L S A P P G 20

28 G A G G G G G G G G G S G G G 19

33 G G G G G G S G G G G A G G G 19

40 G G G G A G G G G G G M Q P G 19

178 P P H Q G H P G G W G A A A A 19

236 A P P G P G G G G G G A G G G 19

290 G P P H H G G G G G G A G P G 19

292 P H H G G G G G G A G P G L N 19

60 S G A Y R G D P S S V K M V Q 18

70 V K M V Q S D F M Q G A M A A 18

74 Q S D F M Q G A M A A S N G G 18

118 A S S P W S G S A V G M A G S 18

119 S S P W S G S A V G M A G S P ^" 18

146 G P D V K G G A G R D D L H A 18

153 A G R D D L H A G T A L H H R 18

204 A H L P S M A G G Q Q P P P Q 18

231 N G M L S A P P G P G G G G G 18

240 P G G G G G G A G G G A Q S L 18

248 G G G A Q S L V H P G L V R G 18

280 P H P P H P H H A Q G P P H H 18

294 H G G G G G G A G P G L N S H 18

11 P G N S L L A A G s I V H S D 17

42 G G A G G G G G G M Q P G S A 17

66 D P S S V K M V Q s D F M Q G 17

80 G M A A S N G G H M L S H A 17

96 Q W V T A L P H A A A A A A A 17

121 P W S G S A V G M A G S P Q Q 17

181 Q G H P G G w G A A A A A A A 17

228 F T V N G M L S A P P G P G G 17

229 T V N G M L S A P P G P G G G 17

242 G G G G G A G G G A Q S L V H 17

243 G G G G A G G G A Q S L V H P 17

7 N P Y L P G N S L L A A G S I 16

13 N S L L A A G S I V H S D A A 16 19 G S I V H S D A A G A G G G G 16 48 G G G M Q P G S A A V T S G A 16 49 G G M Q P G S A A V T S G A Y 16 51 M Q P G S A A V T S G A Y R G 16 57 A V T S G A Y R G D P S s V K 16 75 S D F M Q G A M A A S N G G H 16 87 G G H M L S H A H Q w V T A L 16 97 W V T A L P H A A A A A A A A 16 99 T A L P H A A A A A A A A A A 16 100 A L P H A A A A A A A A A A A 16 101 L P H A A A A A A A A A A A A 16 102 P H A A A A A A A A A A A A V 16 103 H A A A A A A A A A A A A V E 16 104 A A A A A A A A A A A V E A 16 105 A A A A A A A A A A A V E A S 16 107 A A A A A A A A A V E A S S P 16 ^' 116 V E A S S P W S G S A V G M A 16 122 W S G S A V G M A G S P Q Q P 16 124 G S A V G M A G S P Q Q P P Q 16 126 A V G M A G S P Q Q P P Q P P 16 136 P P Q P P P P P P Q G P D V K 16 TABLE XXIII - 187P3F2

(500a. a.) HLA-DRB1*0101 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

138 Q P P P P P P Q G P D V K G G 16 155 R D D L H A G T A L H H R G P 16 165 H H R G P P H L G P P P P P P 16 180 H Q G H P G G W G A A A A A A 16 182 G H P G G W G A A A A A A A A 16 183 H P G G W G A A A A A A A A A 16 185 G G W G A A A A A A A A A A A 16 186 G W G A A A A A A A A A A A A 16 187 W G A A A A A A A A A A A A A 16 188 G A A A A A A A A A A A A A A 16 189 A A A A A A A A A A A A A A 16 190 A A A A A A A A A A A A A A A 16 191 A A A A A A A A A A A A A A H 16 192 A A A A A A A A A A A A H L 16 193 A A A A A A A A A A A A H L P 16 195 A A A A A A A A A A H L P S M 16 196 A A A A A A A A H L P S M A 16 206 L P S M A G G Q Q P P P Q S L 16 214 Q P P P Q S L L Y S Q P G G F 16 296 G G G G G A G P G L N S H D P 16 3 T A A S N P Y L P G N S L L A 15

12 G N S L L A A G s I V H S D A 15

47 G G G G M Q P G s A A V T S G 15

61 G A Y R G D P S s V K M V Q S 15

90 M L S H A H Q W V T A L P H A 15

112 A A A A V E A s s P W S G S A 15

123 S G S A V G M A G s P Q Q P P 15

167 R G P P H L G P P P P P P H Q 15

215 P P P Q S L L Y s Q P G G F T 15

302 G P G L N S H D P H S D E D T 15

15 L L A A G S I V H s D A A G A 14

59 T S G A Y R G D P s S V K M V 14

64 R G D P S S V K M V Q S D F M 14

76 D F M Q G A M A A s N G G H M 14

91 L S H A H Q W V T A L P H A A 14

92 S H A H Q W V T A L P H A A A 14

113 A A A V E A S S P W S G S A V 14

115 A V E A S S P w S G S A V G M 14

125 S A V G M A G s P Q Q P P Q P 14

131 G S P Q Q P P Q P P P P P P Q 14

133 P Q Q P P Q P P P P P P Q G P 14

143 P P Q G P D V K G G A G R D D 14

168 G P H L G P P P P P P H Q G 14

169 P P H L G P P P P P P H Q G H 14

199 A A A A A A H L P S M A G G Q 14

200 A A A A A H L P S M A G G Q Q 14

209 M A G G Q Q P P P Q S L L Y S 14

212 G Q Q P P P Q S L L Y S Q P G 14

230 V N G M L s A P P G P G G G G 14

246 G A G G G A Q s L V H P G L V 14

272 H H H H H H A H P H P P H P H 14

274 H H H H A H P H P P H P H H A 14

277 H A H P H P P H P H H A Q G P 14

308 H D P H S D E D T P T s D D L 14

311 H S D E D T P T s D D L E Q F 14

TABLE XXIV - 187P3F2 (500a . a . ) HLA-DRB1*0301 (DR17 ) 15 - mers

Seq. ID os • 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

19 G S I V H S D A A G A G G G G 22

70 V K M V Q S D F M Q G A M A A 22 57 P G L V R G D T P E L A E H H 22 17 P Q S L L Y S Q P G G F T V N 20 14 E D T P T S D D L E Q F A K Q 20 61 G T A L H H R G P P H L G P P 19 56 H P G L V R G D T P E L A E H ^' 19 26 A V G M A G S P Q Q P P Q P P 18

5 A S N P Y L P G N S L L A A G 17 42 P P P Q G P D V K G G A G R D 17 10 A G G Q Q P P P Q S L L Y S Q 17

88 G H M L S H A H Q W V T A L P 16 06 L P S M A G G Q Q P P P Q S L 16

66 D P S S V K M V Q S D F M Q G 15 47 P D V K G G A G R D D L H A G 15

87 G G H M L S H A H Q W V T A L 14

95 H Q W V T A L P H A A A A A A 14 23 S Q P G G F T V N G M L S A P 14 30 V N G M L S A P P G P G G G G 14

7 N P Y L P G N S L L A A G S I 13

12 G N S L L A A G S I V H S D A 13

48 G G G M Q P G S A A V T S G A 13 51 G G A G R D D L H A G T A L H 13 51 A Q S L V H P G L V R G D T P 13

13 N S L L A A G S I V H S D A A 12

18 A G S I V H S D A A G A G G G 12

55 S A A V T S G A Y R G D P S S 12

60 S G A Y R G D P S S V K M V Q 12

67 P S S V K M V Q S D F M Q G A 12

98 V T A L P H A A A A A A A A A 12 24 G S A V G M A G S P Q Q P P Q 12 69 P P H L G P P P P P P H Q G H 12 18 Q S L L Y S Q P G G F T V N G 12 27 G F T V N G M L S A P P G P G 12 52 Q S L V H P G L V R G D T P E 12 07 S H D P H S D E D T P T s D D 12

69 S V K M V Q S D F M Q G A M A 11

75 S D F M Q G A M A A S N G G H 11 13 A A A V E A S S P W S G S A V 11 46 G P D V K G G A G R D D L H A 11 50 K G G A G R D D L H A G T A L 11 55 R D D L H A G T A L H H R G P 11 03 A A H L P S M A G G Q Q P P P 11 11 G G Q Q P P P Q S L L Y S Q P 11 25 P G G F T V N G M L S A P P G 11 31 N G M L S A P P G P G G G G G 11 64 T P E L A E H H H H H H H H A 11 02 G P G L N S H D P H S D E D T 11 03 P G L N S H D P H S D E D T P 11 09 D P H S D E D T P T S D D L E 11

4 A A S N P Y L P G N S L L A A 10

74 Q S D F M Q G A M A A S N G G 10

79. Q G A M A A S N G G.H M L S H 10

80 G A M A A S N G G H M L S H A 10 53 A G R D D L H A G T A L H H R 10 09 M A G G Q Q P P P Q S L L Y S 10 16 P P Q S L L Y S Q P G G F T V 10 43 G G G G A G G G A Q S L V H P 10 TABLE XXV - 187P3F2 (500a. a. HLA-DRB1*0401 (DR4D 4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

60 S G A Y R G D P S S V K M V Q 28 67 P S S V K M V Q S D F M Q G A 26 88 G H M L S H A H Q W V T A L P 26 95 H Q W V T A L P H A A A A A A 26 124 G S A V G M A G S P Q Q P P Q 26 6 S P Y L P G N S L L A A G S 22

94 A H Q W V T A L P H A A A A A 22

184 P G G w G A A A A A A A A A A 22

12 G N S L L A A G S I V H S D A 20

70 V K M V Q S D F M Q G A M A A 20

75 S D F M Q G A M A A S N G G H 20

87 G G H M L S H A H Q W V T A L 20

98 V A L P H A A A A A A A A A 20

256 H P G L V R G D T P E L A E H 20

257 P G L V R G D T P E L A E H H 20

15 L L A A G S I V H S D A A G A 18

16 L A A G S I V H S D A A G A G 18

51 M Q P G S A A V T S G A Y R G 18

71 K M V Q S D F M Q G A M A A S 18

85 S N G G H M L S H A H Q W V T 18

112 A A A A V E A s S P W S G S A 18

147 P D V K G G A G R D D L H A G 18

151 G G A G R D D L H A G T A L H 18

197 A A A A A A A H L P S M A G 18

248 G G G A Q S L V H P σ L V R G 18

253 S L V H P G L V R G D T P E L 18

308 H D P H s D E D T P T S D D L 18

74 Q S D F M Q G A M A A s N G G 16

119 S S P W S G S A V G M A G S P 16

219 S L L Y S Q P G G F T V N G M 16

225 P G G F T V N G M L S A P P G 16

7 N P Y L P G N S L L A A G S I 14

13 N S L L A A G S I V H S D A A 14

18 A G S I V H S D A A G A G G G 14

19 G S I V H S D A A G A G G G G 14

48 G G G M Q P G S A A V T S G A 14

79 Q G A M A A S N G G H M L S H 14

126 A V G M A G S P Q Q P P Q P P 14

203 A A H L P S M A G G Q Q P P P 14

206 L P S M A G G Q Q P P P Q S L 14

218 Q S L L Y S Q P G G F T V N G 14

227 G F T V N G M L S A P P G P G 14

230 V N G M L S A P P G P G G G G 14

264 T P E L A E H H H H H H H H A 14

TABLE XXVI - 187P3F2

(500a. a.) HLA-DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

95 H Q W V T A L P H A A A A A A 28 158 L H A G T A L H H R G P P H L 23

15 L L A A G S I V H S D A A G A 22 225 P G G F T V N G M L S A P P G 19 227 G F T V N G M L S A P P G P G 18

6 S N P Y L P G N S L L A A G S 17 TABLE XXVI - 187P3F2

(500a. a.) HLA-DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

74 Q S D F M Q G A M A A S N G G 16 94 A H Q W V T A L P H A A A A A 16 119 S S P W S G S A V G M A G S P 16 142 P P P Q G P D V K G G A G R D 16 157 D L H A G T A L H H R G P P H 16 184 P G G W G A A A A A A A A A A 16 248 G G G A Q S L V H P G L V R G 16 253 S L V H P G L V R G D T P E L 16 81 A M A A S N G G H M L S H A H 15 56 A A V T S G A Y R G D P S S V 14 85 S N G G H M L s H A H Q w V T 14 87 G G H M L S H A H Q W V T A L 14 159 H A G T A L H H R G P P H L G 14 163 A L H H R G P P H L G P P P P 14 175 P P P P P H Q G H P G G W G A 14 197 A A A A A A A A H L P S M A G 14 224 Q P G G F T V N G M L s A P P 14 251 A Q S L V H P G L V R G D T P 14 264 T P E L A E H H H H H H H H A 14 267 L A E H H H H H H H H A H P H 14 269 E H H H H H H H H A H P H P P 14 271 H H H H H H H A H P H P P H P 14 279 H P H P P H P H H A Q G P P H 14 285 P H H A Q G P P H H G G G G σ 14 286 H H A Q G P P H H G G G G G G 14 257 P G L V R G D T P E L A E H H 13

TABLE XXIII - 192P2G7 HLA- DRB1*0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

61 I. Q E V V Y L V S Q G A D P D 33

203 L V T M V E Q L A R F L G V S 32

96 L D I I K E L T S P R L I K S 31

250 V G L w K D I F T V S M N E K 31

50 I V T Y P K S G T S L L Q E V 27

111 H L P Y R F L P S D L H N G D 27

163 R R F M N D K L G Y G S W F E 27

16 E S K Y F E F H G V R L P P F 26

45 P S D V W I V T Y P K S G T S 26

63 E V V Y L V S Q G A D P D E I 26

262 N E K F D L V Y K Q K M G K c 26

83 D E Q L P V L E Y P Q P G L D 25

93 Q P G L D I I K E L T S P R L 25

127 K V I Y M A R N P K D L V V S 25

188 D S N V L F L K Y E D M H R D 25

210 L A R F L G V S C D K A Q L E 25

19 Y F E F H G V R L P P F c R G 24

64 V V Y L V S Q G A D P D E I G 24

193 F L K Y E D M H R D L V T M V 24

48 V W I V T Y P K S G T S L L Q 23

102 L T S P R L I K S H L P Y R F 23

105 P R L I K S H L P Y R F L P S 23

118 P S D L H N G D S K V I Y M A 23

200 H R D L V T M V E Q L A R F L 23

32 R G K M E E I A N F P V R P S 22 TABLE XXIII - 192P2G7 HLA- DRB1* 0101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

58 T S L L Q E V V Y L V S Q G A 22

76 Ξ I G L M N I D E Q L P V L E 22

87 P V L E Y P Q P G L D I I K E 22

17 S K F E F H G V R L P P F C 21

77 I G L M N I D E Q L P V L E Y 21

124 G D S K V I Y M A R N P K D L 21

37 E I A N F P V R P S D V W I V 20

79 L M N I D E Q L P V L E Y P Q 20

181 E F W E H R M D S N V L F L K 20

254 K D I F T V S M N E K F D L V 20

266 D L V Y K Q K M G K C D L T F 20

88 V L E Y P Q P G L D I I K E L 19

142 Y Y Q F H R S L R T M S Y R G 19

146 H R S L R T M S Y R G T F Q E 19

155 R G T F Q E F C R R F M N D K 19

249 R V G L W K D I F T V S M N E ^' 19

27 L P P F c R G K M E E I A N F 18

40 N F P V R P S D V W I V T Y P 18

85 Q L P V L E Y P Q P G L D I I 18

99 I K E L T S P R L I K S H L P 18

165 F M N D K L G Y G s W F Ξ H V 18

180 Q E F W E H R M D s N V L F L 18

256 I F T V S M N E K F D L V Y K 18

24 G V R L P P F C R G K M E E I 17

35 M E E I A N F P V R P S D V w 17

46 S D V w I V T Y P K S G T S L 17

49 W I V T Y P K s G T S L L Q E 17

54 P K S G T S L L Q E V V Y L V 17

106 R L I K s H L P Y R F L P S D 17

140 V S Y Y Q F H R S L R T M S Y 17

196 Y E D M H R D L V T M V E Q L 17

201 R D L V T M V E Q L A R F L G 17

217 S C D K A Q L E A L T E H C H 17

223 L E A L T E H C H Q L V D Q C 17

234 V D Q C C N A E A L P V G R G 17

240 A E A L P V G R G R V G L W K 17

246 G R G R V G L W K D I F T V S 17

265 F D L V Y K Q K M G K C D L T 17

267 L V Y K Q K M G K C D L T F D 17

9 P S T P G E F E S K Y F E F H 16

31 C R G K M E E I A N F P V R P 16

38 I A F P V R P S D V W I V T 16

56 S G T S L L Q E V V Y L V S Q 16

57 G T S L L Q E V V Y L V s Q G 16

95 G L D I I K Ξ L T S P R L I K 16

97 D I I K E L T S P R L I K s H 16

101 E L T s P R L I K S H L P Y R 16

115 R F L P s D L H N G D S K V I 16

128 V I Y M A R N P K D L V V s Y 16

137 D L V V S Y Y Q F H R S L R T 16

143 Y Q F H R s L R T M S Y R G T 16

173 G S W F E H V Q E F W E H R M 16

176 F E H V Q E F W E H R M D s N 16

197 E D M H R D L V T M V E Q L A 16

204 V T M V E Q L A R F L G V S C 16

207 V E Q L A R F L G V S C D K A 16

213 F L G V S C D K A Q L E A L T 16

220 K A Q L E A L T E H C H Q L V 16 TABLE XXIII - 192P2G7 HLA- DRB1*0101 15 - mers

Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

H Q L V D Q C C N A E A L P V^' 16 Q L V D Q C C N A E A L P V G 16

TABLE XXIV - 192P2G7 HLA- DRB1*0301 (DR17) 15 - mers

Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

F W E H R M D S N V L F L K Y 29

I G L M N I D E Q L P V L E Y 28

Q L P V L E Y P Q P G L D I I 28

S D L H N G D S K V I Y M A R 26

I F T V S M N E K F D L V Y K 26

K D I F T V S M N E K F D L V 24

Y R F L P S D L H N G D S K V 23

F L G V S C D K A Q L E L T 23

P R L I K S H L P Y R F L P S 21

Y E D M H R D L V T M V E Q L 21

R D L V T M V E Q L A R F L G 21

L V T M V E Q L A R F L G V S 21

L D I I K E L T S P R L I K S 20

M A R N P K D L V V S Y Y Q F 20

R V G L w K D I F T V S M N E 20

T V S M N Ξ K F D L V Y K Q K 20

Q E V V Y L V S Q G A D P D E 19

Y P Q P G L D I I K E L T S P 19

C R R F M N D K L G Y G S W F 19

L F L K Y E D M H R D L V T M 19

H R D L V T M V E Q L A R F L 19

G V R L P P F C R G K M E Ξ I 18

D V W I V T Y P K S G T S L L 18

E I G L M N I D E Q L P V L E 18

Q P G L D I I K E L T S P R L 18

D S K V I Y M A R N P K D L V 18

S K V I Y M A R N P K D L V V 18

V I Y M A R N P K D L V V S Y 18

K D L V V S Y Y Q F H R S L R 18

R G K M E E I A N F V R P S 17

F E H V Q E F W E H R M D S N 17

S N V L F L K Y E D M H R D L 17

V T M V E Q L A R F L G V S C 17

R F L G V S C D K A Q L E A L 17

H Q L V D Q C.C N A E A L P V 17

E A L P V G R G R V G L W K D 17

N E K F D L V Y K Q K M G K C 17

K F D L V Y K Q K M G K C D L 17

Y F E F H G V R L P P F C R G 16

K S G T S L L Q E V V Y L V S 16

R S L R T M S Y R G T F Q E F 16

Q E F C ^•R R F M N D K L σ Y G 16

F C R R F M N D K L G Y G S W 16

K A Q L E A L T E H C H Q L V 16

T P S T P G E F E S K Y F E F 15

Y Y Q F H R S L R T M S Y R G 15

R G T F Q E F C R R F M N D K 15 ^' G R G R V G L W K D I F T V s 15

L V Y K Q K M G K C D L T F D 15

V S Q G A D P D E I G L M N I 14 TABLE XXIV - 192P2G7 HLA- DRB1*0301 (DR17) 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

101 E L T S P R L I K S H L P Y R 14 173 G S W F E H V Q E F W E H R M 14 223 L E A L T E H C H Q L V D Q C 14

TABLE XXV - 192P2G7 HLA- DRB1*0401 (DR4D 4) 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

155 R G T F Q Ξ F C R R F M N D K 28

250 V G L W K D I F T V S M N E K 28

24 G V R L P P F C R G K M E E I 26

32 R G K M E E I A N F P V R P s 26

61 L Q E V V Y L V S Q G A D P D 26

77 I G L M N I D E Q L P V L Ξ Y 26

96 L D I I K E L T S P R L I K s 26

16 E S K Y F E F H G V R L P P F 22

50 I V T Y P K S G T S L L Q E V 22

111 H L P Y R F L P s D L H N G D 22

139 V V s Y Y Q F H R S L R T M S 22

140 V S Y Y Q F H R S L R T M S Y 22

142 Y Y Q F H R S L R T M S Y R G 22

193 F L K Y E D M H R D L V T M V 22

210 L A R F L G V S C D K A Q L E 22

262 N E K F D L V Y K Q K M G K C 22

35 M E E I A N F P V R P S D V W 20

45 P S D V W I V T Y P K S G T s 20

48 V W I V T Y P K S G T S L L Q 20

57 G T s L L Q E V V Y L V S Q G 20

58 T S L L Q E V V Y L V s Q G A 20

62 Q E V V Y L V s Q G A D P D E 20

85 Q P V L E Y P Q P G L D I I 20

93 Q P G L D I I K Ξ L T S P R L 20

95 G L D I I K E L T S P R L I K 20

114 Y R F L P s D L H N G D S K V 20

118 P S D L H N G D S K V I Y M A 20

125 D S K V I Y M A R N P K D L V 20

126 S K V I Y M A R N P K D L V V 20

176 F E H V Q E F W E H R M D S N 20

188 D S N V L F L K Y E D M H R D 20

196 Y E D M H R D L V T M V E Q L 20

201 R D L V T M V E Q L A R F L G 20

204 V T M V E Q L A R F L G V S C 20

223 L E A L T E H C H Q L V D Q C 20

231 H Q L V D Q C C N A E A L P V 20

249 R V G L W K D I F T V S M N E 20

253 W K D I F T V s M N E K F D L 20

44 R P S D V w I V T Y P K S G T 18

54 P K S G T s L L Q E V V Y L V 18

115 R F L P S D L H N G D s K V I 18

133 R N P K D L V V S Y Y Q F H R 18

159 Q E F C R R F M N D K L G Y G 18

178 H V Q E F W E H R M D S N V L 18

192 L F L K Y E D M H R D L V T M 18

228 E H C H Q L V D Q C C N A E A 18

239 N A E A L P V G R G R V G L W 18

19 Y F E F H G V R L P P F C R G 16

27 L P P F C R G K M E E I A N F 16

88 V L E Y P Q P G L D I I K E L 16 TABLE XXV - 192P2G7 HLA- DRB1*0401 (DR4DW4) 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

151 T M S Y R G T F Q E F C R R F 16 162 C R R F M N D K L G Y G s W F 16 169 K L G Y G S W F E H V Q E F W 16 172 Y G S W F E H V Q E F W E H R 16 173 G S W F Ξ H V Q E F W Ξ H R M 16 179 V Q E F W E H R M D S N V L F 16 254 K D I F T V S M N E K F D L V 16 40 N F P V R P S D V W I V T Y P 14 47 D V W I V T Y P K S G T s L L 14 74 P D E I G L M N I D E Q L P V 14 76 E I G L M N I D E Q L P V L E 14 83 D E Q L P V L E Y P Q P G L D 14 99 I K E L T S P R L I K s H L P 14 128 V I Y M A R N P K D L V V S Y 14 136 K D L V V S Y Y Q F H R s L R 14 137 D L V V S Y Y Q F H R s L R T 14 146 H R S L R T M S Y R G T F Q E 14 184 E H R M D S N V L F L K Y E D 14 191 V L F L K Y E D M H R D L V T 14 200 H R D L V T M V E Q L A R F L 14 203 L V T M V E Q L A R F L G V S 14 207 V E Q L A R F L G V S C D K A 14 211 A R F L G V S C D K A Q L E A 14 213 F L G V S C D K A Q L E A L T 14 220 K A Q L E A L T E H C H Q L V 14 247 R G R V G L W K D I F T V S M 14 256 I F T V S M N E K F D L V Y K 14

TABLE XXVI - 192P2G7 HLA- DRB1*1101 15 - mers

Seq. ID

Pos 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

204 V T M V E Q L A R F L G V S C 27

193 F L K Y E D M H R D L V T M V 25

262 N E K F D L V Y K Q K M G K C 25

142 Y Y Q F H R S L R T M S Y R G 24

210 L A R F L G V S C D K A Q L E 23

58 T S L L Q E V V Y L V S Q G A 21

200 H R D L V T M V Ξ Q L A R F L 21

61 L Q E V V Y L V S Q G A D P D 20

125 D S K V I Y M A R N P K D L V 20

45 P S D V w I V T Y P K S G T S 19

76 E I G L M N I D E Q L P V L E 19

83 D E Q L P V L E Y P Q P G L D 19

139 V V S Y Y Q F H R S L R T M S 19

155 R G T F Q E F C R R F M N D K 19

17 S K Y F E F H G V R L P P F C 18

93 Q P G L D I I K E L T S P R L 18

18 K Y F E F H G V R L P P F C R 17

46 S D V W I V T Y P K s G T S L 17

63 E V V Y L V S Q G A D P D E I 17

111 H L P Y R F L P S D L H N G D 17

223 L E A L T E H C H Q L V D Q C 17

19 Y F Ξ F H G V R L P P F C R G 16

113 P Y R F L P S D L H N G D S K 16

162 C R R F M N D K L G Y G S W F 16

173 G S W F E H V Q E F W E H R M 16

239 N A E A L P V G R G R V G L W 16 TABLE XXVI - 192P2G7 HLA- DRB1*1101 15 - mers

Pos Seq. ID

1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 score Num.

250 V G L W K D I F T V S M N E K 16

24 G V R L P P F C R G K M E E I 15

92 P Q P G L D I I K E L T S P R 15

99 I K E L T S P R L I K S H L P 15

107 L I K S H L P Y R F L P S D L 15

128 V I Y M A R N P K D L V V S Y 15

178 H V Q E F W E H R M D s N V L 15

240 A E A L P V G R G R V G L W K 15

246 G R G R V G L W K D I F T V S 15

22 F H G V R L P P F C R G K M E 14

47 D V W I V T Y P K S G T S L L 14

48 V W I V T Y P K s G T s L L Q 14

96 L D I I K E L T s P R L I K S 14

101 E L T S P R L I K S H L P Y R 14

114 Y R F L P S D L H N G D s K V 14

137 D L V V S Y Y Q F H R S L R T 14

201 R D L V T M V E Q L A R F L G 14

237 C C N A E A L P V G R G R V G 14

254 K D I F T V S M N E K F D L V 14

256 I F T V S M N E K F D L V Y K 14

264 K F D L V Y K Q K M G K C D L 14

267 L V Y K Q K M G K C D L T F D 14

16 E S K Y F E F H G V R L P P F 13

35 M E E I A N F P V R P S D V W 13

102 L T S P R L I K S H L P Y R F 13

146 H R S L R T M S Y R G T F Q E 13

160 E F C R R F M N D K L G Y G S 13

172 Y G S W F E H V Q E F W Ξ H R 13

249 R V G L W K D I F T V S M N E 13

253 W K D I F T V s M N E K F D L 13

Claims (51)

CLAIMS:

1. A composition comprising: a substance that a) modulates the status of a protein of Figure 2 (SEQ ID NOS: ), or b) a molecule that is modulated by a protein of Figure 2, whereby the status of a cell that expresses a protein of Figure 2 is modulated.

2. A composition of claim 1 , further comprising a physiologically acceptable carrier.

3. A pharmaceutical composition that comprises the composition of claim 1 in a human unit dose form.

4. A composition of claim 1 wherein the substance comprises an antibody or fragment thereof that specifically binds to a protein that is related to a protein of Figure 2.

5. An antibody or fragment thereof of claim 4, which is monoclonal.

6. An antibody of claim 4, which is a human antibody, a humanized antibody or a chimeric antibody.

7. A non-human transgenic animal that produces an antibody of claim 4.

8. A hybridoma that produces an antibody of claim 5.

9. A method of delivering a cytotoxic agent or a diagnostic agent to a cell that expresses a protein of Figure 2 (SEQ ID NOS: ), said method comprising: providing the cytotoxic agent or the diagnostic agent conjugated to an antibody or fragment thereof of claim 4; and, exposing the cell to the antibody-agent or fragment-agent conjugate.

10. A composition of claim 1 wherein the substance comprises a polynucleotide that encodes an antibody or fragment thereof, either of which immunospecifically bind to a protein of Figure 2.

11. A composition of claim 1 wherein the substance comprises a protein related to a protein of Figure

12. A protein of claim 11 that is at least 90% homologous to an entire amino acid sequence shown in Figure 2 (SEQ ID NOS:

13. A composition of claim 1 wherein the substance comprises: a) a peptide of eight, nine, ten, or eleven contiguous amino acids of a protein of Figure 2; b) a peptide of Tables V to XVIII (SEQ ID NOS: ); or, c) a peptide of Tables XXIII to XXVI (SEQ ID NOS: ).

14. A composition of claim 1 wherein the substance comprises a CTL polypeptide or an analog thereof, from the amino acid sequence of a protein of Figure 2 (SEQ ID NOS: ).

15. A composition of claim 14 further limited by a proviso that the epitope is not an entire amino acid sequence of Figure 2 (SEQ ID NOS:).

16. A composition of claim 14 wherein the substance comprises a CTL polypeptide set forth in Tables V to XVIII (SEQ ID NOS: ).

17. A composition of claim 16 further limited by aproviso that the polypeptide is not an entire amino acid sequence of a protein of Figure 2 (SEQ ID NOS: ).

18. A composition of claim 1 wherein the substance comprises an antibody polypeptide epitope from an amino acid sequence of Figure 2 (SEQ ID NOS: ).

19. A composition of claim 18 further limited by a proviso that the epitope is not an entire amino acid sequence of Figure 2 (SEQ ID NOS: ).

20. A composition of claim 18 wherein the antibody epitope comprises a peptide region of at least 5 amino acids of Figure 2 (SEQ ID NOS: ) in any whole number increment up to the end of said peptide, wherein the epitope comprises an amino acid position selected from: a) an amino acid position having a value greater than 0.5 in the Hydrophilicity profile of Figure 5, b) an amino acid position having a value less than 0.5 in the Hydropathicity profile of Figure 6; c) an amino acid position having a value greater than 0.5 in the Percent Accessible Residues profile of Figure 7; d) an amino acid position having a value greater than 0.5 in the Average Flexibility profile of Figure 8; e) an amino acid position having a value greater than 0.5 in the Beta-turn profile of Figure 9; f) ₍ a combination of at least two of a) through e); g) a combination of at least three of a) through e); h) a combination of at least four of a) through e); or, i) a combination of five of a) through e).

21. A composition of claim 20 further limited by a proviso that the epitope is not an entire amino acid sequence of Figure 2 (SEQ ID NOS: ).

22. A polynucleotide that encodes a protein of claim 11.

23. A polynucleotide of claim 22 that comprises a nucleic acid molecule set forth in Figure 2.

24. A polynucleotide of claim 22 further limited by a proviso that the encoded protein is not an entire amino acid sequence of Figure 2 (SEQ ID NOS: ).

25. A polynucleotide of claim 22 wherein T is substituted with U.

26. A composition of claim 1 wherein the substance comprises a polynucleotide that comprises a coding sequence of a nucleic acid sequence of Figure 2 (SEQ ID NOS: ).

27. A polynucleotide of claim 22 that further comprises an additional nucleotide sequence that encodes an additional a protein related to a protein of Figure 2.

28. A composition comprising a polynucleotide that is fully complementary to a polynucleotide of claim 22.

29. A composition comprising a polynucleotide that is fully complementary to a polynucleotide of claim 25.

30. A composition comprising a polynucleotide that is fully complementary to a polynucleotide of claim 27.

31. A composition of claim 1 wherein the substance comprises a) a ribozyme that cleaves a polynucleotide having a protein of Figure 2 coding sequence, or b) a nucleic acid molecule that encodes the ribozyme; and, a physiologically acceptable carrier.

32. A composition of claim 1 wherein the substance comprises human T cells, wherein said T cells specifically recognize a protein of Figure 2 peptide subsequence in the context of a particular HLA molecule.

33. A method of inhibiting growth of cancer cells that express a protein of Figure 2, the method comprising: administering to the cells the composition of claim 1.

34. A method of claim 33 of inhibiting growth of cancer cells that express a protein of Figure 2, the method comprising steps of: administering to said cells an antibody or fragment thereof, either of which specifically bind to a protein of Figure 2-related protein.

35. A method of claim 33 of inhibiting growth of cancer cells that express a protein of Figure 2, the method comprising steps of: administering to said cells a protein of Figure 2-related protein.

36. A method of claim 33 of inhibiting growth of cancer cells that express a protein of Figure 2, the method comprising steps of: administering to said cells a polynucleotide comprising a coding sequence for a protein of Figure 2-related protein or comprising a polynucleotide complementary to a coding sequence for a protein of Figure 2-related protein.

37. A method of claim 33 of inhibiting growth of cancer cells that express a protein of Figure 2, the method comprising steps of: administering to said cells a ribozyme that cleaves a polynucleotide that encodes a protein of Figure 2.

38. A method of claim 33 of inhibiting growth of cancer cells that express a protein of Figure 2 and a particular HLA molecule, the method comprising steps of: administering human T cells to said cancer cells, wherein said T cells specifically recognize a peptide subsequence of a protein of Figure 2 while the subsequence is in the context ofthe particular HLA molecule.

39. A method of claim 33, the method comprising steps of: administering a vector that delivers a nucleotide that encodes a single chain monoclonal antibody, whereby the encoded single chain antibody is expressed intracellularly within cancer cells that express a protein of Figure 2.

40. A method of generating a mammalian immune response directed to a protein of Figure 2, the method comprising: exposing cells ofthe mammal's immune system to a portion of a) a protein of Figure 2-related protein and/or b) a nucleotide sequence that encodes said protein, whereby an immune response is generated to said protein.

41. A method of generating an immune response of claim 40, said method comprising: providing a protein of Figure 2-related protein that comprises at least one T cell or at least one B cell epitope; and, contacting the epitope with a mammalian immune system T cell or B cell respectively, whereby the T cell or B cell is activated.

42. A method of claim 41 wherein the immune system cell is a B cell, whereby the induced B cell generates antibodies that specifically bind to the protein of Figure 2-related protein.

43. A method of claim 41 wherein the immune system cell is a T cell that is a cytotoxic T cell (CTL), whereby the activated CTL kills an autologous cell that expresses the protein of Figure 2-related protein.

44. A method of claim 41 wherein the immune system cell is a T cell that is a helper T cell (HTL), whereby the activated HTL secretes cytokines that facilitate the cytotoxic activity of a cytotoxic T cell (CTL) or the antibody-producing activity of a B cell.

45. A method for detecting, in a sample, the presence of a protein of Figure 2-related protein or a polynucleotide of Figure 2-related polynucleotide, comprising steps of: contacting the sample with a substance of claim 1 that specifically binds to the protein of Figure 2-related protein or to the polynucleotide of Figure 2-related polynucleotide, respectively; and, determining that there is a complex ofthe substance with the protein of Figure 2-related protein or the substance with the protein of Figure 2-related polynucleotide, respectively.

46. A method of claim 45 for detecting the presence of a protein of Figure 2-related protein in a sample comprising steps of: contacting the sample with an antibody or fragment thereof either of which specifically bind to the protein of Figure 2-related protein; and, deteπnining that there is a complex ofthe antibody or fragment thereof and the protein of Figure 2-related protein.

47. A method of claim 45 further comprising a step of: taking the sample from a patient who has or who is suspected of having cancer.

48. A method of claim 45 for detecting the presence of a protein of Figure 2 mRNA in a sample comprising: producing cDNA from the sample by reverse transcription using at least one primer; amplifying the cDNA so produced using a protein of Figure 2 polynucleotides as sense and antisense primers, wherein the protein of Figure 2 polynucleotides used as the sense and antisense primers serve to amplify a protein of Figure 2 cDNA; and, detecting the presence ofthe amplified protein of Figure 2 cDNA.

49. A method of claim 45 for monitoring at least one protein of Figure 2 gene product in a biological sample from a patient who has or who is suspected of having cancer, the method comprising: deteiroining the status of a nucleotide of Figure 2 gene product expressed by cells in a tissue sample from an individual; comparing the status so determined to the status ofthe nucleotide of Figure 2 gene product in a coπesponding normal sample; and, identifying the presence of aberrant gene products ofthe nucleotide of Figure 2 in the sample relative to the normal sample.

50. The method of claim 49 further comprising a step of deteπnining if there are elevated gene products of a nucleotide of Figure 2, mRNA or a protein, whereby the presence of elevated gene products in the test sample relative to the normal tissue sample indicates the presence or status of a cancer.

51. A method of claim 50 wherein the cancer occurs in a tissue set forth in Table I.