T CELL RECEPTOR-BASED THERAPY FOR RHEUMATOID ARTHRITIS

Abstract: There is provided by this invention a novel method of treating rheumatoid arthritis in a mammal. The method comprises the steps of obtaining a sample of synovium from the mammal; identifying in said sample T cell receptor variable regions; and administering to said mammal an effective amount of antibodies to at least one of said T cell receptor variable regions or antigenic fragments thereof. The invention further provides a novel method of treating rheumatoid arthritis in a mammal comprising the steps of administering to said mammal an effective amount of antibodies to mammalian T cell receptor variable regions selected from the group consisting of V alpha 17, V alpha 1, V beta 12, V beta 14, V beta 17 and V beta 7 and antigenic fragments thereof. The invention further comprises a novel method for immunizing a mammal to prevent the occurrence of rheumatoid arthritis or to treat ongoing rheumatoid arthritis. The method comprises the steps of administering to said mammal mammalian T cell receptor variable regions selected from the group consisting of V alpha 17, V alpha 1, V beta 12, V beta 14, V beta 17, V beta 7 and antigenic fragments thereof. Kits comprising mammalian T cell receptor variable regions selected from the group consisting of V alpha 17, V alpha 1, V beta 12, V beta 14, V beta 17 and V beta 7 and antigenic fragments thereof or antibodies to said variable regions are also provided by the invention.

Description: T CELL RECEPTOR-BASED THERAPY FOR RHEUMATOID ARTHRITIS Cross Reference to Related Application This application is a continuation-in-part of U.S. Serial No. 750,913 entitled "T Cell Receptor-Based Therapy for Rheumatoid Arthritis" filed in the U.S. Patent and Trademark Office on August 28, 1991 which is incorporated by reference herein. Field of the Invention This invention relates to the field of mammalian therapeutics. More particularly, methods of treating rheumatoid arthritis and methods for immunizing against, rheumatoid arthritis are provided. Government Rights The work presented herein was supported in part by National Institute of Health grant 1R-29AI-28503-01. The United States Government has certain rights in the invention. Background of the invention Rheumatoid arthritis (RA) is a systemic polyarthropathy characterized pathologically by proliferation of synovial fibroblast-like and macrophage-like cells and infiltration of the synovium with lymphocytes, predominately T cells of the helper (CD4+) phenotype (1,2). Such CD4+ T cells are typically activated by an antigenic peptide complexed with Class II MHC molecules (HLA-DR/DP/DQ) . Immunogenetic analysis reveals that RA is associated with HLA- DR4, and more specifically with glutamine/lysine residues at amino acids 70/71 of the HLA-DR/3 chain (3-8) . Current therapy for rheumatoid arthritis is either poorly efficacious or toxic. Many lines of evidence indicate that T cells are involved in the development of rheumatoid joint disease. This includes the presence of lymphocytic infiltrates composed primarily of CD4+ T cells in the synovium (2, 21-23) the linkage of RA to HLA-DR4 which comprises a ligand for CD4+ T cell antigen receptors (3-8) , and experimental models of arthritis and related autoimmune diseases which can be transferred by T cell lines (10, 13, 24- 33) . Studies in both animal models and human rheumatoid arthritis indicate that anti-T cell reagents can be of therapeutic efficacy (11, 25, 34-40) . However, if these reagents are non-specific and delete too large a portion of the T cell repertoire, immunodeficiency (such as seen in acquired immune deficiency syndrome or AIDS) may result. A better therapeutic alternative is to delete only those T cells involved in the autoimmune response. Since these comprise only a small portion of the total T cell repertoire, eliminating these T cells should not result in significant generalized immunosuppression. Summary of the invention There is provided by this invention a novel method of treating rheumatoid arthritis in a mammal. The method comprises the steps of obtaining a sample of synovium from the mammal; identifying in said sample T cell receptor variable regions; and administering to said mammal an effective amount of antibodies to at least one of said T cell receptor variable regions or antigenic fragments thereof. The invention further provides a novel method of treating rheumatoid arthritis in a mammal comprising the steps of administering to said mammal an effective amount of antibodies to mammalian T cell receptor variable regions selected from the group consisting of Vαl7, Vαl, V/312, V|8l4, V/317 and Vβl and antigenic fragments thereof. The invention further comprises a novel method for immunizing a mammal to prevent the occurrence of rheumatoid arthritis or to treat ongoing rheumatoid arthritis. The method comprises the steps of administering to said mammal mammalian T cell receptor variable regions selected from the group consisting of Vαl7, Vαl, V/312, V314, V/317, V/37 and antigenic fragments thereof. Kits useful in the methods of the present invention comprising mammalian T cell receptor variable regions selected from the group consisting of Vαl7, Vαl, V/312, V/314, V/317 and V/37 and antigenic fragments thereof or antibodies to said variable regions are also provided by the invention. Rheumatoid arthritis (RA) is characterized by massive proliferation of synovial tissue, elevated expression of HLA DR antigens, accompanying infiltration of the tissue with CD4+ T lymphocytes, and a genetic linkage to the major histocompatility (MHC) antigen HLA-DR4. Since T cells are restricted by Class II MHC molecules such as DR4, this suggests a direct role for these CD4+ cells in pathogenesis. One strategy for the development of novel therapies in T cell mediated autoimmunity is to specifically delete the autoreactive T cells. Such a strategy depends on understanding the molecular structure of autoreactive T cell receptors (TCR) . To investigate the TCR usage in RA, oligonucleotide primers specific for each of the major TCR subfamilies - one set for the TCR alpha chains and one for the TCR beta chains were used. These were utilized to amplify cDNA derived from whole synovium or synovial tissue T cell lines in a family specific manner. Amplified cDNA was sequenced to determine the corresponding amino acid sequences. Detection of amplified DNA was facilitated by utilizing oligonucleotide probes derived from the constant regions of the TCRs. Synovial T cell lines were developed by stimulation with phytohemagglutinin followed by maintenance in IL-2. The TCR repertoire present in these cell lines was quite heterogeneous, with an average of 15 alpha chains and 15.8 beta chains detected. When synovial tissue was analyzed, the predominant TCR subfamilies detected tended to be more restricted, with an average of 4.2 alpha chains and 9.7 beta chains detected. In some synovial tissue samples predominance of one subfamily was apparent. These results suggest that while a polyclonal population of T cells is present in RA synovium, the predominant patterns of TCR transcript expression may be somewhat more restricted. This suggests that TCR based therapy of RA is possible. Brief Description of the Drawings Figure 1. T cell receptor specific oligonucleotides and their relative location. Figure 2. TCR transcripts in RA synovial T cell lines. Rheumatoid synovial T cell lines were developed by initial culture in PHA for 3-5 days, then maintained in IL-2 at 10 U/ml. Following 1-3 weeks of passage, the cells were frozen, and RNA later extracted for analysis of TCR expression as outlined in Materials andMethods. The sample designations are shown on the left, with the corresponding TCR alpha and beta family-specific primers used indicated above each lane. Figure 3. TCR transcripts in RA synovium. RNA was extracted and cDNA synthesized form 10 rheumatoid synovial tissues obtained at the time of joint surgery. These were analyzed for TCR expression as noted above. The sample designations are shown on the left, with the corresponding TCR alpha and beta family-specific primers used indicated above each lane. Figure 4. (A) Graphic representation of the frequency of occurrence of individual alpha chain variable regions in rheumatoid synovial tissue and T cell lines; (B) Graphic representation of the frequency of occurrence of individual beta chain variable regions in rheumatoid synovial tissue and T cell lines. Figure 5. T cell receptor PCR primers. The asterisk denotes antisense primer. C/3 1 and C/3 2 primers were used mixed together in equimolar concentrations. Figure 6. T cell receptor β chain expression in ten rheumatoid synovia, The asterisk denotes > 2 standard errors from the mean. Figure 7. T cell receptor α chain expression in ten rheumatoid synovia, The asterisk denotes > 2 standard errors from the mean. Detailed Description of the Invention In one aspect of the invention a method of treating rheumatoid arthritis in a mammal, such as a human, is provided. The method comprises obtaining a sample of synovium from the mammal; identifying in said sample T cell receptor variable regions; and administering to said mammal an effective amount of antibodies to at least one of said T cell receptor variable regions or antigenic fragments thereof. Samples of synovium such as synovial tissue or fluid are obtained as is known to those in the art. Molecular characterization of human T cell receptors has been greatly aided recently through the application of the polymerase chain reaction (PCR). (19) See also e. g. U.S. patent 4,386,202 issued to Mullis which patent is incorporated by reference as if fully set forth herein. By utilizing oligonucleotide primers specific for the different T cell receptor variable region families, family specific amplification is possible (14-16) . This technique can conveniently be applied to the identification of T cell receptors of interest. Sequences of T cell receptors are generally available in the literature and in computer-based sequence data bases such as "Genbank" and "EMBL". Thus, the sequence of the family-specific oligonucleotide primer of interest can be matched against these data bases utilizing a variety of computer software tools (For example, the University of Wisconsin package. (49)) with programs such as "Word Search and Segments" or "Best Fit". The matched sequence are retrieved from the data base and translated from nucleic acid to protein sequence. Alternatively, the T cell receptors of interest can be identified by in situ hybridization. Northern or Southern blot analysis of synovial fluid or tissue with family-specific probes or by immunohistochemistry or immunofluorescence with antibodies to the various T cell receptor variable regions, where available. An effective amount of antibodies to at least one of the T cell receptor variable regions is then administered to the mammal. It should be noted that "antibodies to at least one of the T cell receptor variable regions" is meant to denote antibodies which recognize T cell receptor variable regions and portions or fragments thereof. An effective amount of antibodies is that amount which reduces the level of T cells bearing the corresponding receptor in the synovium or which results in clinical signs of improvement in the patient. An antibody is said to be "capable of binding" a molecule if it is capable of specifically reacting with the molecule to thereby bind the molecule to the antibody. The term "epitope" is meant to refer to that portion of an antigen which can be recognized and bound by an antibody. An antigen may have one or more than one epitope. An "antigen" is a substance capable of inducing an animal to produce antibodies capable of binding to an epitope of that antigen. The specific reaction referred to above is meant to indicate that the antigen will immunoreact, in a highly selective manner, with its corresponding antibody and not with the multitude of other antibodies which may be evoked by other antigens. The term "antibody" (Ab) or "monoclonal antibody" (Mab) as used herein is meant to include intact molecules as well as fragments thereof (such as, for example. Fab and F(ab') 2 fragments) which are capable of binding an antigen. Fab and F(ab , 2 ) fragments lack the Fc fragment of intact antibody, clear more rapidly from the circulation, and may have less non-specific tissue binding. The antibodies useful in the present invention may be prepared by any of a variety of methods. Antibodies useful in the present invention include antibodies to the T cell receptor variable region as well as antibodies to antigenic fragments thereof. Methods for the production of such antibodies are well known and described fully in the literature. (19) For example, cells expressing the peptide, synthetic peptides or an antigenic fragment thereof, can be administered to an animal in order to induce the production of sera containing polyclonal antibodies that are capable of binding the peptide. Peptides useful in the present invention may range in size from about 25 to about 500 amino acids in length. In some embodiments of the present invention peptides may be from about 50 to about 300 amino acids in length. In still other embodiments of the present invention peptides may be from about 50 to about 200 amino acids in length. Generally, a peptide fragment is prepared and purified to render it substantially free of natural contaminants or a peptide fragment is synthesized, according to means known in the art. Either the purified fragment or the synthesized fragment or a combination of purified natural fragments and/or synthesized fragment may be introduced into an animal in order to produce polyclonal antisera of greater specific activity. Monoclonal antibodies can be prepared using known hybridoma technology. In general, such procedures involve immunizing an animal with a peptide antigen, which includes the T cell receptor variable region and antigenic fragments thereof. The splenocytes of such animals are extracted and fused with a suitable myeloma cell line. Any suitable myeloma cell line may be employed in accordance with the present invention. After fusion, the resulting hybridoma cells are selectively maintained in a suitable medium and then cloned by limiting dilution. The hybridoma cells obtained through such a selection are then assayed to identify clones which secrete antibodies capable of binding the peptide antigen. If the peptide source is impure, only some of the hybridoma cells will produce antibodies capable of binding to the peptide (other hybridoma cells will produce antibody capable of binding to the peptide contaminants) . Thus, it may be necessary to screen among the hybridoma cells for those which are capable of secreting an antibody which is capable of binding to the peptide. Once such a hybridoma cell has been identified, it may be clonally propagated by means known in the art in order to produce the peptide-specific monoclonal antibody. The sequence of many human T cell receptor variable regions are known and are available in data bases such as "Gen Bank" and "EMBL". Additional sequences of interest may be determined by cloning and sequencing cDNA clones of T cell receptors isolated from synovial tissue or fluid (48) . In particular sequences of T cell receptor variable regions V014, V/317, Vαl and Vαl7 are preferred. Preferred DNA sequences and corresponding amino acid sequences of these regions are set forth in Table 1. Table 1 sets forth preferred sequences of rheumatoid synovial T cell receptor α and β chain variable regions derived from human synovial tissue. Such sequences and portions of said sequences are useful for the development of antibodies useful in the present invention. It should be understood by those skilled in the art that, in some embodiments of the present invention nucleic acid analogs may be substituted for naturally occuring nucleic acids. In preferred embodiments of the present invention nucleic acid sequences may range from about 75 to about 1500 nucleic acid bases in length based upon the portion of the T cell receptor variable region being coded and the size of a particular T cell receptor variable region. In other preferred embodiments nucleic acid sequences may range in length from about 150 to about 900 nucleic acid bases. In yet other embodiments of the present invention from about 150 to about 600 nucleic acids may code for a selected T cell receptor variable region or portion thereof. Table I I. Rheumatoid Synovial T Cell Receptor Bet Nucleic Acid and Amino Acid Sequence 1. Patient 6, Clone /314.1 (SEQ ID NO: 1) Family Specific Primer V/314 GTG ACT GAT AAG GGA GAT GTT CCT GAA GGG TAC AGT GTC TC Val Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Se 1 5 10 AAG AAG GAG CGC TTC TCC CTG ATT CTG GAG TCC GCC AGC AC Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Th 20 25 30 ACA TCT ATG TAC CTC TGT GCC AGC AGT TCA CAA AAA CCC AA Thr Ser Met Tyr Leu Cys Ala Ser Ser Ser Gin Lys Pro As 35 40 45 ACC TTC GGT TCG GGG ACC AGG TTG TCC GTT GTA GAG GAC CT Thr Phe Gly Ser Gly Thr Arg Leu Ser Val Val Glu Asp L 50 55 " 60 GTG TTC CCA CCC GAG GTC GCT GTG TTT GAG CCA TCA GAA G Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu A 65 70 75 2. Patient 6, Clone 014.2 (SEQ ID NO:3) Family Specific Primer V314 GTG ACT GAT AAG GGA GAT GTT CCT GAA GGG TAC AGT GTC TCT AGA GAG Val Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Ser Arg Glu 1 5 10 15 AAG AAG GAG CGC TTC TCC CTG ATT CTG GAG TCC GCC AGC ACC AAC CAG Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 ACA TCT ATG TAC CTC TGT GCC AGC AGT TGG GGG ACT GAA GCT TTC TTT Thr Ser Met Tyr Leu Cys Ala Ser Ser Trp Gly Thr Glu Ala Phe Phe 35 40 45 GGA CAA GGC ACC AGA CTC ACA GTT GTA GAG GAC CTG AAC AAG GTG TTC Gly Gin Gly Thr Arg Leu Thr Val Val Glu Asp Leu Asn Lys Val Phe 50 55 60 CCA CCC GAG GTC GCT GTG TTT GAG CCA TCA GAA GCA GAG Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 3. Patient 5, Clone 014.3/4/5/6 (SEQ ID NO:5) Family Specific Primers V/314 GAG ACT GAT AAG GGA GAT GTT CCT GAA GGG TAC AGT GTC TCT AGA GAG Glu Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Ser Arg Glu 1 5 10 15 AAG AAG GAG CGC TTC TCC CTG ATT CTG GAG TCC GCC AGC ACC AAC CAG Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 ACA TCT ATG TAC CTC TGT GCC AGC AGT TTG CTC CAG CGG ACC ACC ACA Thr Ser Met Tyr Leu Cys Ala Ser Ser Leu Leu Gin Arg Thr Thr Thr 35 40 45 ' r GAT ACG CAG TAT TTT GGC CCA GGC ACC CGG CTG ACA GTG CTC GAG GAC Asp Thr Gin Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu Glu Asp ' 50 55 60 CTG AAA AAC GTG TTC CCA CCC GAG ATC GCT GTG TTT GAG CCA TCA GAA Leu Lys Asn Val Phe Pro Pro Glu He Ala Val Phe Glu Pro Ser Glu 65 70 75 80 GCA GAG Ala Glu 4. Patient 5, Clone 014.7 (SEQ ID NO:7) Family Specific Primer V/314 GTG ACT GAT AAG GGA GAT GTT CCT GAA GGG TAC AGT GTC TCT AGA GAG Val Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Ser Arg Glu 1 5 10 15 AAG AAG GAG CGC TTC TCC CTG ATT CTG GAG TCC GCC AGC ACC AAC CAG Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 ACA TCT ATG TAC CTC TGT GCC AGC AGC CTG GAC AGG GGC GAG CAG TAC Thr Ser Met Tyr Leu Cys Ala Ser Ser Leu Asp Arg Gly Glu Gin Tyr 35 40 45 ' TTC GGG CCG GGC ACC AGG CTC ACG GTC ACA GAG GAC CTG AAA AAC GTG Phe Gly Pro Gly Thr Arg Leu Thr Val Thr Glu Asp Leu Lys Asn Val 50 55 60 TTC CCA CCC GAG GTC GCT GTG TTT GAG CCA TCA GAA GCA GAG Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 5. Patient 5, clone /314.8 (SEQ ID NO:9) Family Specific Primer V/314 GTG ACT GAT AAG GGA GAT GTT CCT GAA GGG TAC AGT GTC TCT AGA GAG Val Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Ser Arg Glu 10 15 AAG AAG GAG CGC TTC TCC CTG ATT CTG GAG TCC GCC AGC ACC AAC CAG Lys Lys Glu Arg Phe Ser Leu He, Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 ACA TCT ATG TAC CTC TGT GCC AGC AGT TTA ACC TCC GTC ACA GAT ACG Thr Ser Met Tyr Leu Cys Ala Ser Ser Leu Thr Ser Val Thr Asp Thr 35 40 45 I CAG TAT TTT GGC CCA GGC ACC CGG CTG ACA GTG CTC GAG GAC CTG AAA Gin Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu Glu Asp Leu Lys 50 55 60 ' AAC GTG TTC CCA CCC GAG GTC GCT GTG TTT GAG CCA TCA GAA GCA GAG Asn Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 80 6. Patient 6, Clone /317.1 (SEQ ID NO:11) Family Specific Primer V/317 TTT CAG AAA GGA GAT ATA GCT GAA GGG TAC AGC GTC TCT CGG GAG AAG Phe Gin Lys Gly Asp He Ala Glu Gly Tyr Ser Val Ser Arg Glu Lys 1 5 10 15 AAG GAA TCC TTT CCT CTC ACT GTG ACA TCG GCC CAA AAG AAC CCG ACA Lys Glu Ser Phe Pro Leu Thr Val Thr Ser Ala Gin Lys Asn Pro Thr 20 25 30 GCT TTC TAT CTC TGT GCC AGT AGT ATT GGG GGA CAA GGG CTA ACC GGG Ala Phe Tyr Leu Cys Ala Ser Ser He Gly Gly Gin Gly Leu Thr Gly 35 40 45 , GCC AAA AAC ATT CAG TAC TTC GGC GCC GGG ACC CGG CCC TCA GTG CTG υl Ala Lys Asn He Gin Tyr Phe Gly Ala Gly Thr Arg Pro Ser Val Leu , 50 55 60 GAG GAC CTG AAA AAC GTG TTC CCA CCC GAG GTC GCT GTG TTT GAG CCA Glu Asp Leu Lys Asn Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro 65 70 75 80 TCA GAA GCA GAG Ser Glu Ala Glu 7. Patient 6, Clone 017.2 (SEQ ID NO:13) Family specific Primer V/317 TTT CAG AAA GGA GAT ATA GCT GAA GGG TAC AAA GTC TCT CGA AAA GAG Phe Gin Lys Gly Asp He Ala Glu Gly Tyr* Lys Val Ser Arg Lys Glu 1 5 10 15 AAG AGG AAT TTC CCC CTG ATC CTG GAG TCG CCC AGC CCC AAC CAG ACC Lys Arg Asn Phe Pro Leu He Leu Glu Ser Pro Ser Pro Asn Gin Thr 20 25 30 TCT CTG TAC TTC TGT GCC AGC AGT TTG GGG GGA ACC TAC AAT GAG CAG Ser Leu Tyr Phe Cys Ala Ser Ser Leu Gly Gly Thr Tyr Asn Glu Gin 35 40 45 TTC TTC GGG CCA GGG ACA CGG CTC ACC GTG CTA GAG GAC CTG AAA AAC Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu Glu Asp Leu Lys Asn 50 55 60 GTG TTC CCA CCC GAG GTC GCT GTG TTT GAG CCA TCA GAA GCA GAG Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 8. Patient 6, Clone 017.3 (SEQ ID NO: 33) Family Specific Primer V/317 TTT CAG AAA GGA GAT ATA GCT GAA GGG TAC AAA GTC TCT CGA AAA GAG Phe Gin Lys Gly Asp He Ala Glu Gly Tyr Lys Val Ser Arg Lys Glu 1 5 10 15 AAG AGG AAT TTC CCC CTG ATC CTG GAG TCG CCC AGC CCC AAC CAG ACC Lys Arg Asn Phe Pro Leu He Leu Glu Ser Pro Ser Pro Asn Gin Thr 20 25 30 TCT CTG TAC TTC TGT GCC AGC AGT CCG TTC TCT CGA GCA TCC TAT GGC Ser Leu Tyr Phe Cys Ala Ser Ser Pro Phe Ser Arg Ala Ser Tyr Gly 35 40 45 TAC ACC TTC GGT TCG GGG AAC AGG TTA ACC GTT GTA GAG GAC CTG AAA Tyr Thr Phe Gly Ser Gly Asn Arg Leu Thr Val Val Glu Asp Leu Lys 50 55 60 AAC GTG TTC CCA CCC GAG GTC GCT GTG TTT GAG CCA TCA GAA GCA GAG Asn Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 80 II. Rheumatoid. Synovial T Cell Receptor Alpha Chains Nucleic Acid and Amino Acid Sequences 9. Patient 1, Clone αl.l/2 (SEQ ID NO:15) Family Specific Primer Vαl CTG AGG TGC AAC TAT TCC TAT GGG GCA ACA CCT TAT CTC TTC TGG TAT Leu Arg Cys Asn Tyr Ser Tyr Gly Ala Thr Pro Tyr Leu Phe Trp Tyr 1 5 10 15 GTC CAG TCC CCC GGC CAA GGC CTC CAG CTG CCC CTG AAG TAC TTT TCA Val Gin Ser Pro Gly Gin Gly Leu Gin Leu Pro Leu Lys Tyr Phe Ser 20 25 30 GGA GAC ACT CTG GTT CAA GGC ATT AAA GGC TTT GAG GCT GAA TTT AAG Gly Asp Thr Leu Val Gin Gly He Lys Gly Phe Glu Ala Glu Phe Lys ' 35 40 45 ι AGG AGT CAA TCT TCC TTC AAT CTG AGG AAA CCC TCT GTG CAT TGG AGT ' Arg Ser Gin Ser Ser Phe Asn Leu Arg Lys Pro Ser Val His Trp Ser 50 55 60 GAT GCT GCT GAG TAC TTC TGT GCT GTG GGT GCT GAT TCA GGA TAC AGC Asp Ala Ala Glu Tyr Phe Cys Ala Val Gly Ala Asp Ser Gly Tyr Ser 65 70 75 80 ACC CTC ACC TTT GGG AAG GGG ACT ATG CTT CTA GTC TCT CCA GAT ATC Thr Leu Thr Phe Gly Lys Gly Thr Met Leu Leu Val Ser Pro Asp He 85 90 95 CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC TCT AAA TCC AGT Gin Asn Pro GAC AAG Asp Lys 10. Patient 2, Clone αl.3 (SEQ ID NO:17) Family Specific Primer Vαl CTG AGG TGC AAC TAT TCC TAT GGG GCA ACA CCT TAT CTC TTC TGG TAT Leu Arg Cys Asn Tyr Ser Tyr Gly Ala GTC CAG TCC CCC GGC CAA GGC CTC CAG CTG CTC CTG AAG TAC TTT TCA Val Gin Ser Pro Gly Gin Gly Leu Gin Leu Leu Leu Lys Tyr Phe Ser 20 25 30 GGA GAC ACT CTG GTT CAA GGC ATT AAA GGC TTT GAG GCT GAA TTT AAG Gly Asp Thr Leu Val Gin Gly He Lys Gly Phe Glu Ala Glu Phe Lys 35 40 45 AGG AGT CAA TCT TCC TTC AAT CTG AGG AAA CCC TCT GTG CAT TGG AGT Arg Ser Gin Ser Ser Phe Asn Leu Arg Lys Pro Ser Val His Trp Ser 50 55 60 GAT GCT GCT GAG TAC TTC TGT GCT GTG GGT CCC ACC CAC AAT GAC ATG Asp Ala Ala Glu Tyr Phe Cys Ala Val Gly Pro Thr His Asn Asp Met 65 70 75 80 CGC TTT GGA GCA GGG ACC AGA CTG ACA GTA AAA CCA AAT ATC CAG AAC Arg Phe Gly Ala Gly Thr Arg Leu Thr Val Lys Pro Asn He Gin Asn 85 90 95 CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC TCT AAA TCC AGT GAC AAG Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp Ser Lys Ser Ser Asp Lys 100 105 110 11. Patient 2, Clone αl.4 (SEQ ID NO: 19) Family Specific Primer Vαl CTG AGG TGC AAC TAT TCC TAT TCT GGG AGT CCT GAA CTC TTC TGG TAT Leu Arg Cys Asn Tyr Ser Tyr Ser Gly Ser Pro Glu Leu Phe Trp Tyr 1 5 10 15 GTC CAG TAC TCC AGA CAA CGC CTC CAG TTA CTC TTG AGA CAC ATC TCT Val Gin Tyr Ser Arg Gin Arg Leu Gin Leu Leu Leu Arg His He Ser 20 25 30 AGA GAG AGC ATC AAA GGC TTC ACT GCT GAC CTT AAC AAA GGC GAG ACA Arg Glu Ser He Lys Gly Phe Thr Ala Asp Leu Asn Lys Gly Glu Thr 35 40 45 TCT TTC CAC CTG AAG AAA CCA TTT GCT CAA GAG GAA GAC TCA GCC ATG Ser Phe His Leu Lys Lys Pro Phe Ala Gin Glu Glu Asp Ser Ala Met 50 55 60 ' TAT TAC TGT GCT CTA GCG CTG CAG GCA ACA AGC TTA CTT TTG GAG GAG Tyr Tyr Cys Ala Leu Ala Leu Gin Ala Thr Ser Leu Leu Leu Glu Glu 65 70 75 80 GAA CCC AGG GTG CTA GTT AAA CCA AAT ATC CAG AAC CCT GAC CCT GCC Glu Pro Arg Val Leu Val Lys Pro Asn He Gin Asn Pro Asp Pro Ala 85 90 95 GTG TAC CAG CTG AGA GAC TCT AAA TCC AGT GAC AAG Val Tyr Gin Leu Arg Asp Ser Lys Ser Ser Asp Lys 100 105 12. Patient 2, Clone αl7.1 (SEQ ID NO: 25) Family Specific Primer Vαl7 C TTG TCA CTG GAT TTA GAG TCT CTC AGC TGC AGT TAC ACA GTC AGC Leu Ser Leu Asp Leu Glu Ser Leu Ser Cys Ser Tyr Thr Val Ser 1 5 10 15 GGT TTA AGA GGG CTG TTC TGG TAT AGG CAA GAT CCT GGG AAA GGC CCT Gly Leu Arg Gly Leu Phe Trp Tyr Arg Gin Asp Pro Gly Lys Gly Pro 20 25 30 GAA TTC CTC TTC ACC CTG TAT TCA GCT GGG GAA GAA AAG GAG AAA GAA Glu Phe Leu Phe Thr Leu Tyr Ser Ala Gly Glu Glu Lys Glu Lys Glu 35 40 45 AGG CTA AAA GCC ACA TTA ACA AAG AAG GAA AGC TTT CTG CAC ATC ACA Arg Leu Lys Ala Thr Leu Thr Lys Lys Glu Ser Phe Leu His He Thr 50 55 60 GCC CCT AAA CCT GAA GAC TCA GCC ACT TAT CTC TGT GCT GTG AGG CGA Ala Pro Lys Pro Glu Asp Ser Ala Thr Tyr Leu Cys Ala Val Arg Arg 65 70 75 TCA GAT GGC CAG AAG CTG CTC TTT GCA AGG GGA ACC ATG TTA AAG GTG Ser Asp Gly Gin Lys Leu Leu Phe Ala Arg Gly Thr Met Leu Lys Val 80 85 90 95 GAT CTT AAT ATC CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC Asp Leu Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp 100 105 110 TCT AAA TCC AGT GAC AAG Ser Lys Ser Ser Asp Lys 115 13. Patient 3, Clone αl7.2 (SEQ ID NO:23) Family Specific Primer Vαl7 C TTG TCA CTG GAT TTA GAG TCT CTC AGC TGC AGT TAC ACA GTC AGC Leu Ser Leu Asp Leu Glu Ser Leu Ser Cys Ser Tyr Thr Val Ser 1 5 10 15 GGT TTA AGA GGG CTG TTC TGG TAT AQG CAA GAT CCT GGG AAA GGC CCT Gly Leu Arg Gly Leu Phe Trp Tyr Arg Gin Asp Pro Gly Lys Gly Pro 20 25 30 GAA TTC CTC TTC ACC CTG TAT TCA GCT GGG GAA GAA AAG GAG AAA GAA Glu Phe Leu Phe Thr Leu Tyr Ser Ala Gly Glu Glu Lys Glu Lys Glu 35 40 45 AGG CTA AAA GCC ACA TTA ACA AAG AAG GAA AGC TTT CTG CAC ATC ACA Arg Leu Lys Ala Thr Leu Thr Lys Lys Glu Ser Phe Leu His He Thr 50 55 60 GCC CCT AAA CCT GAA GAC TCA GCC ACT TAT CTC TGT GCT GTG AGG CGA Ala Pro Lys pro Glu Asp Ser Ala Thr Tyr Leu Cys Ala Val Arg Arg 65 70 75 TCA GAT GGC CAG AAG CTG CTC TTT GCA AGG GGA ACC ATG TTA AAG GTG Ser Asp Gly Gin Lys Leu Leu Phe Ala Arg Gly Thr Met Leu Lys Val 80 85 90 95 GAT CTT AAT ATC CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC Asp Leu Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp 100 105 110 TCT AAA TCC AGT GAC AAG Ser Lys Ser Ser Asp Lys 115 14. Patient 4, clone αl7.3 (SEQ ID NO: 21) Family Specific Primer Vαl7 C TTG TCA CTG GAT TTA GAG TCT CTC AGC TGC AGT TAC ACA GTC AGC Leu Ser Leu Asp Leu Glu Ser Leu Ser Cys Ser Tyr Thr Val Ser 1 5 10 15 GGT TTA AGA GGG CTG TTC TGG TAT AGG CAA GAT CCT GGG AAA GGC CCT Gly Leu Arg Gly Leu Phe Trp Tyr Arg Gin Asp Pro Gly Lys Gly Pro 20 25 30 GAA TTC CTC TTC ACC CTG TAT TCA GCT GGG GAA GAA AAG GAG AAA GAA Glu Phe Leu Phe Thr Leu Tyr Ser Ala Gly Glu Glu Lys Glu Lys Glu 35 40 45 AGG CTA AAA GCC ACA TTA ACA AAG AAG GAA AGC TTT CTG CAC ATC ACA Arg Leu Lys Ala Thr Leu Thr Lys Lys Glu Ser Phe Leu His He Thr 50 55 60 ' GCC CCT AAA CCT GAA GAC TCA GCC ACT TAT CTC TGT GCT GCG AGG CGA Ala Pro Lys Pro Glu Asp Ser Ala Thr Tyr Leu Cys Ala Ala Arg Arg 65 70 75 TCA GAT GGC CAG AAG CTG CTC TTT GCA AGG GGA ACC ATG TTA AAG GTG Ser Asp Gly Gin Lys Leu Leu Phe Ala Arg Gly Thr Met Leu Lys Val 80 85 90 95 GAT CTT AAT ATC CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC Asp Leu Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp 100 105 110 TCT AAA TCC AGT GAC AAG Ser Lys Ser Ser Asp Lys 15. Patient A , Clone αl7.4 (SEQ ID NO: 27) Family Specific Primer Vαl7 C TTG TCA CTG GAT TTA GAG TCT CTC AGC TGC AGT TAC ACA GTC AGC Leu Ser Leu Asp Leu Glu Ser Leu Ser Cys Ser Tyr Thr Val Ser 1 5 10 15 GGT TTA AGA TGG CTG TTC TGG TAT AGG CAA GAT CCT GGG AAA GGC CCT Gly Leu Arg Trp Leu Phe Trp Tyr Arg Gin Asp Pro Gly Lys Gly Pro 20 25 30 GAA TTC CTC TTC GCC CTG TAT TCA GCT GGG GAA GAA AAG GAG AAA GAA Glu Phe Leu Phe Ala Leu Tyr Ser Ala Gly Glu Glu Lys Glu Lys Glu 35 40 45 AGG CTA AAA GCC ACA TTA ACA AAG AAG GAA AGC TTT CTG CAC ATC ACA Arg Leu Lys Ala Thr Leu Thr Lys Lys Glu Ser Phe Leu His He Thr 50 55 60 GCC CCT AAA CCT GAA GAC TCA GCC ACT TAT CTC TGT GCT GTG AGG CGA Ala Pro Lys Pro Glu Asp Ser Ala Thr Tyr Leu Cys Ala Val Arg Arg 65 70 75 TCA GAT GGC CAG AAG CTG CTC TTT GCA AGG GGA ACC ATG TTA AAG GTG Ser Asp Gly Gin Lys Leu Leu Phe Ala Arg Gly Thr Met Leu Lys Val 80 85 90 95 GAT CTT AAT ATC CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC Asp Leu Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp 100 105 110 TCT AAA TCC AGT GAC AAG Ser Lys Ser Ser Asp Lys 115 16. Patient 7, clone αl7.5 (SEQ ID NO: 29) Family Specific Primer Vαl7 A GAT GTC TCC ATG AAC TGC ACT TCT TCA AGC ATA TTT AAC ACC TGG Asp Val Ser Met Asn Cys Thr Ser Ser Ser He Phe Asn Thr Trp 1 5 10 15 CTA TGG TAC AAG CAG GAC CCT GGG GAA GGT CCT GTC CTC TTG ATA GCC Leu Trp Tyr Lys Gin Asp Pro Gly Glu Gly Pro Val Leu Leu He Ala 20 25 30 TTA TAT AAG GCT GGT GAA TTG ACC TCA AAT GGA AGA CTG ACT GCT CAG Leu Tyr Lys Ala Gly Glu Leu Thr Ser Asn Gly Arg Leu Thr Ala Gin 35 40 45 TTT GGT ATA ACC AGA AAG GAC AGC TTC CTG AAT ATC TCA GCA TCC ATA Phe Gly He Thr Arg Lys Asp Ser Phe Leu Asn He Ser Ala Ser He 50 55 60 CCT AGT GAT GTA GGC ATC TAC TTC TGT GCT GGG CAG GCC CTC ACC GGT Pro Ser Asp Val Gly He Tyr Phe Cys Ala Gly Gin Ala Leu Thr Gly 65 70 75 AAC CAG TTC TAT TTT GGG ACA GGG ACA AGT TTG ACG GTC ATT CCA AAT Asn Gin Phe Tyr Phe Gly Thr Gly Thr Ser Leu Thr Val He Pro Asn 80 85 90 95 ATC CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC TCT AAA TCC He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp Ser Lys Ser 100 105 110 17. Patient 4, Clone αl7.6 (SEQ ID NO:31) Family Specific Primer Vαl7 CTT GTC ACT GGA TTT AGA GTC TCT CAG CTG GTG GAG CAG AGC CCT CAA Leu Val Thr Gly Phe Arg Val Ser Gin Leu,Val Glu Gin Ser Pro Gin 1 5 10 15 TCT TTG ATA GTC CAG AAA GGA GGG ATT TCA ATT ATA AAC TGT GCT TAT Ser Leu He Val Gin Lys Gly Gly He Ser He He Asn Cys Ala Tyr 20 25 30 GAG AAC ACT GCG TTT GAC TAC TTT CCA TGG TAC CAA CAA TTC CCT GGG Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr Gin Gin Phe Pro Gly 35 40 45 I AAA GGC CCT GCA TTA TTG ATA GCC ATA CGT CCA GAT GTG AGT GAA AAG Lys Gly Pro Ala Leu Leu He Ala He Arg Pro Asp Val Ser Glu Lys 50 55 60 ' AAA GAA GGA AGA TTC ACA ATC TCC TTC AAT AAA AGT GCC AAG CAG TTC Lys Glu Gly Arg Phe Thr He Ser Phe Asn Lys Ser Ala Lys Gin Phe 65 70 75 80 TCA TTG CAT ATC ATG GAT TCC CAG CCT GGA GAC TCA GCC ACC TAC TTC Ser Leu His He Met Asp Ser Gin Pro Gly Asp Ser Ala Thr Tyr Phe 85 90 95 TGT GCA GCA GAG GGA GGA AAG CTT ATC TTC GGA CAG GGA ACG GAG TTA Cys Ala Ala Glu Gly Gly Lys Leu He Phe Gly Gin Gly Thr Glu Leu 100 105 110 TCT GTG AAA CCC AAT ATC CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG Ser Val Lys Pro Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu 115 120 125 AGA GAC TCT AAA TCC AGT GAC AAG Arg Asp Ser Lys Ser Ser Asp Lys 130 135 Antibodies may be developed against the T cell receptors or against amino acid sequences and portions thereof, corresponding to said T cell receptor variable regions such as those set forth in Table 1 for commercial purposes by developing monoclonal antibodies as indicated herein and known in the art. These murine or rat or other species monoclonals could be administered directly. Alternatively, to reduce xenogeneic responses to the monoclonals, these antibodies can be "humanized" by grafting a human constant region onto the non-human variable region, or by transplanting the non-human hypervariable regions onto a human antibody. (50, 51) Polyclonal antibodies can also be employed, particularly if they are from a species which exhibits little immunogenicity in humans such as pigs. Antigenic fragments may be derived from family-specific sequences such as those contained in the variable region primers or from hypervariable regions as defined in Jones et al. (52) The association of RA with HLA-is reminiscent of similar associations seen in experimental models of autoimmunity, such as experimental autoimmune encephalomyelitis, a model for multiple sclerosis triggered by autoreactive T cells reactive to myelin basic protein and specific MHC Class II antigens (9-12) . The observation of a restriction to certain MHCs in such experimental systems correlates with a restricted repertoire of T cell antigen receptors which respond to that MHC + antigen (13) . This has also been documented in multiple sclerosis T cell lines derived from humans (14, 15). In experimental systems, antibodies directed to the relevant T cell receptors, or immunization with peptides derived from these T cell receptors, is capable of ameliorating the disease (10, 11) . In another embodiment of the invention a method of treating rheumatoid arthritis in a mammal is provided which comprises administering to said mammal an effective amount of antibodies to mammalian T cell receptor variable regions selected from the group consisting of Vαl7 r Vαl, V/312, V/314, V/317 and V37 and antigenic fragments thereof. In particular, antibodies to amino acid set forth in Table 1 and portions thereof are preferred. Antibodies to mammalian T cell receptor variable regions selected from the group consisting of Vαl7, Vαl, V/312, V/314, V/317 and V/37 and antigenic fragments thereof can be prepared as described above. An effective amount of antibodies to at least one of the T cell receptor variable regions described above is then administered to the mammal. An effective amount of antibodies is that amount which reduces the level of T cells bearing the corresponding receptor in the synovium or which results in clinical signs of improvement in the patient. Of course the method of treating rheumatoid arthritis of the present invention may be combined with other traditional treatments for the disease where indicated. It is believed the therapy of the invention could be administered at any point in the course of rheumatoid arthritis. A method for immunizing a mammal to prevent the occurrence of rheumatoid arthritis or to ameliorate active disease is also provided by the invention. The method comprises administering to said mammal mammalian T cell receptor variable regions selected from the group consisting of Vαl7, Vαl, V/312, V017, Vβl and antigenic fragments thereof. Amino acid sequences as set forth in Table 1, and portions thereof, are preferred for some embodiments of the invention. Mammals could be immunized by using the T cell receptor variable regions described above and antigenic fragments thereof, with or without agents known to those in the art attached thereto to increase the antigenic potential of the antigen. Generally the antigen or protein can be dissolved at between about lμg/ml to about lg/ml in sterile saline or saline with 0.4 mg aluminum hydroxide per ml as a vehicle. Generally 0.5 to 1.0 ml of the protein solution is injected intramuscularly and then followed by booster injections at one and 6-12 months after the initial immunization. An effective amount is that amount of antigen sufficient to raise antibodies to the antigen in the animal. There is precedence for immunizing mammals with T cell receptor variable regions as protection against experimental autoimmune encephalomyelitis. (11, 12) It is believed that a patient to be immunized would either have clinical evidence of rheumatoid arthritis, have a strong family history of rheumatoid arthritis or have the genetic predisposition for rheumatoid arthritis described herein. Kits with the antibodies described herein useful in the treatment of rheumatoid arthritis or kits with antigens for immunization are also within the scope of this invention. Materials and Methods Synovial tissue and Cell Lines: Tissue was obtained at the time of joint surgery, and was handled sterily at all times. The tissue was rinsed in sterile phosphate buffered saline (PBS) , placed in a petri dish, the superficial layer snipped off with scissors and minced with a sterile scalpel. The minced tissue was placed in 20 mis PBS with 5% HEPES buffer, 0.4 g hyaluronidase (type 1-S) , 0.04 g DNA-ase 1 (type II from bovine pancrease) and 1.2 g collagenase (Type Z) (all from Sigma, St. Louis, MO) with 1% fetal calf serum (FCS) , and stirred continuously for 90 minutes at 37°C. The large chunks of tissue were decanted, and the cells centrifuged and washed twice in culture media (RPMI 1640 with pen/step, L-glutamine, sodium pyruvate, non-essential amino acids, HEPES buffer 5X10 " M /3-mercaptoethanol (all from Gibco, Gaithersburg MD) , and 10% FCS (Hyclone) . The T cells were purified by standard nylon wool chromatography (17) , cultured overnight at 1x10 /ml in culture media, and the non-adherent cells separated, centrifuged, and maintained in culture. Stimulation of the cells was with either phytohemagglutinin (1% solution, from Sigma) , interleukin-2 (Amgen Biologicals, Thousand Oaks, CA) , or media alone. Cells were stimulated for 3-5 days, and then maintained for varying periods of time in 10 U/ml IL-2 prior to analysis. Fluorescence-Activated Cell Sorter (FACS) Analysis: Following culture, cells were centrifuged, washed and resuspended in FACS media (1% bovine serum albumin in PBS with 0.1% sodium azide) , at lxlO 6 cells per lOOμl. Primary antibody was added for 20-40 minutes on ice. After an additional two washings, the cells were subjected to second antibody (fluorescein isothiocyanate-conjugated goat anti-mouse Ig (Sigma); at 1:100 dilution), then washed twice again. The cells were then analyzed at the University of Pennsylvania Cancer Center FACS facility. Per cent positive was determined by comparing the samples to a no primary antibody control. Antibodies used were 0KT3 anti-CD3 (Ortho Diagnostics, Raritan, NJ) , Leu3a anti-CD4 (Becton-Dickinson provide location) , and OKT8 (Ortho) , at the dilutions suggested by the suppliers. RNA Extraction and cDNA Synthesis: Tissue was homogenized in guanidinum isothiocyanate (GITC) solution, or cells resuspended in GITC solution, and vortexed for 30 seconds. 0.1 ml 2 M sodium acetate pH 4 was added, the solution vortexed, followed by 1.0 ml diethylpyrocarbonate (DEP)-^water-saturated phenol, the sample mixed, then 0.2 ml phenylisoamyl alcohol, thorough vortexing, and the solution transferred to sterile EPPENDORF tubes. Each sample was then incubated on ice for 20 minutes, microfuged for 10 minutes, and the top layer recovered, RNA precipitated with 2.5 volumes of 100% ethanol and 1/10 volume 1M sodium acetate pH 5.5 in dry ice/ethanol for 30 minutes. The solutions were microfuged for 15 minutes, the supernatant decanted, the pellets washed in 70% ethanol and rotary evaporated. The dried pellets were resuspended in 50 μl DEP-water and RNA quantitated spectrophotometrically. For reverse transcription, 1-20 μg of RNA in lOμl was utilized to synthesize cDNA primed with random hexamers iϊi the following reactionmixture: 3μl MaloneyMurine Leukemia Virus reverse transcriptase with 6 μl 5x reverse transcriptase buffer, 1.5 μl RNAse inhibitor, and 3 μl 0.1 M dithiothreitol (all from GIBCO/BRL, Gaithersburg, MD) , 3 μl random hexamers (fromPharmacia LKB Biotechnology, Piscataway, NJ) , and either 1 or 3 μl 100 M dNTPs (25 mM in each dNTP, from Boehringer Mannheim, GmbH W. Germany) . Following a 10 minute preincubation at 25°C, the reaction was carried out for 1 hour at 42°C, then 95°C for 5 minutes followed by storage at -20°C until use. PCR Amplification T Cell Receptor Variable Regions: cDNA was amplified utilizing the primers listed in Figure 5 with Va/jSn and Ca/β mid at 0.2 nM concentrations. cDNA was amplified utilizing Tήeπαus aguaticus DNA polymerase (Tag polymerase) and standard reaction conditions suggested by themanufacturer (Perkin-Elmer Cetus Corp., Norwalk, CT) . The reaction mixture contained 10 μl of 10X reaction buffer, 16 μl 1.25 nM dNTPs (final concentration 200 jiM in each dNTP) , 5 μl of each oligonucleotide primer at 20 ^ (final 1 JXM in each primer) , 5 μl of DNA, 0.5 μl of DNA, 0.5 μl Tag polymerase, and 58.5 ml distilled/deionized water. Primers were synthesized by the Wistar Institute oligonucleotide synthesis facility. The program utilized 5 initial low temperature cycles for low stringency (95°C for 1 min., 37°C for 2 min., 52°C for 2 min.), followed by higher stringency for 40 cycles (95°C for 1 min., 52°C for 2 min., 72°C for 2 min) , and a final 5 minute 72°C elongation phase. For some experiments, the initial 20 cycles, described above, was used followed by additional increments of 5 higher stringency cycles (95°C for 1 min., 52°C for 2 min., 72°C for 2 min), with PCR product removed following each increment of 5 cycles for analysis. Products were analyzed by electrophoresis on 2-3% agarose gels stained with ethidium bromide. Determination of sequences of T Cell Receptor Variable Regions: PCR products were cloned into the TA cloning vector (InVitrogen, San Diego, CA) according to kit instructions. Plasmid DNA was isolated from the clones as described by Ausubel, et al.. Current Protocols in Molecular Biology (John Wiley & Sons, New York, NY) and Sambrook, et al. , Molecular Cloning. A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.) incorporated by reference in their entireties. A portion of the cloned cDNA was sequenced in accordance with methods provided by Ausubel, et al., Current Protocols in Molecular Biology (John Wiley & Sons, New York, NY) and Sambrook, et al., Molecular Cloning. A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.) incorporated by reference in their entireties. Amino acid sequences were determined as set forth in Table 1. Relative positions set forth in Table 2 were determined in relation to family specific variable region primers used and published data providing invariant residues τ/*ε 68Z_.0/Z6Sfl/LDd S69IO/£6 OΛ\ RHEUMATOID SYNOVIAL T CELL RECEPTOR BETA CHAINS CONSERVED RESIDUES V Q P I G I S V T POSITION (Includes Leader Peptide) 1 10 20 30 40 50 60 70 Patient 6 Clone 014.1 (SEQ ID NO:l) V . DKG Patient 6 Clone 014.2 (SEQ ID NO:3) VT . DKG Patient 5 Clone 014.3/4/5/6 (SEQ ID NO:5) ET . DKG ω ■ Patient 5 Clone 014.7 (SE ID NO:7) VT . DKG IO Patient 5 Clone 014.8 (SEQ ID NO:9) VT. DKG Patient 6 Clone 017.1 (SEQ ID NO:11) FQ . KG Patient 6 Clone 017.2 (SEQ ID NO:13) FQ . . KG Patient 6 Clone 017.3 (SEQ ID NO:33) FQ. KG TABLE II RHEUMATOID SYNOVIAL T CELL RECEPTOR ALPHA CHAINS CONSERVED FG RESIDUES A POSITION (Includes Leader Peptide) 80 90 100 110 120 tfe. Patient 1 Clone αl.1/2 (SEQ ID NO: 15) KGFEAEFKRSQSSFNLRKPSVHWSDAAEYFCAV...GADSGYSTLTFGKG ω Patient 2 Clone αl.3 (SEQ ID NO:17) KGFEAEFKRSQSSFNLRKPSVHWSDAAEYFCAV...GP..THNDMRFGAG Patient 2 Clone αl.4 (SEQ ID NO:19) FTADLNKGETS...FHLKKPFAQEEDSAMYYCAL...ALQATSLLLEEEP Patient 2 Clone αl7.1 (SEQ ID NO:25) ERLKATLTKKESFLHITAPKPE..DSATYLCAVRRSDGQKLLFARGTMLK Patient 3 Clone αl7.2 (SEQ ID NO:23) ERLKATLTKKESFLHITAPKPE..DSATYLCAVRRSDGQKLLFARGTMLK Patient 4 Clone αl7.3 (SEQ ID NO:21) ERLKATLTKKESFLHITAPKPE..DSATYLCAARRSDGQKLLFARGTMLK Patient 4 Clone αl7.4 (SEQ ID NO:27) ERLKATLTKKESFLHITAPKPE..DSATYLCAVRRSDGQKLLFARGTMLK Patient 7 Clone αl7.5 (SEQ ID NO:29) RLTAQFGITRKDSFLNISASIP.SDVGIYFCAGQALTGNQFYFGTGTSLT Patient 4 Clone αl7.6 (SEQ ID NO:31) GRFTISFNKSAKQFSLHIMDSQPGDSATYFCAAEGGKLI...FGQGTELS RHEUMATOID SYNOVIAL T CELL RECEPTOR BETA CHAINS CONSERVED RESIDUES POSITION (Includes Leader Peptide) 80 90 100 110 Patient 6 Clone 014.1 (SEQ ID NO:l) DVPEGYSVSREKKERFSLILESASTNQTSMYLCASSSQKP Patient 6 Clone 014.2 (SEQ ID NO:3) DVPEGYSVSREKKERFSLILESASTNQTSMYLCASSWGT. Patient 5 Clone 014.3/4/5/6 (SEQ ID NO: 5) DVPEGYSVSREKKERFSLILESASTNQTSMYLCASSLLQR ω Patient 5 Clone 014.7 (SE ID NO:7) DVPEGYSVSREKKERFSLILESASTNQTSMYLCASSLDRG Patient 5 Clone 014.8 (SEQ ID NO:9) DVPEGYSVSREKKERFSLILESASTNQTSMYLCASSLTSV Patient 6 Clone 017.1 (SEQ ID NO:11) DIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASSIGGQ Patient 6 Clone 017.2 (SEQ ID NO:13) DIAEGYKVSRKEKRNFPLILESPSPNQTSLYFCASSLGGT Patient 6 Clone 017.3 (SEQ ID NO:33) DIAEGYKVSRKEKRNFPLILESPSPNQTSLYFCASSPFSR TABLE II RHEUMATOID SYNOVIAL T CELL RECEPTOR ALPHA CHAINS CONSERVED RESIDUES NIQ D Y POSITION (Includes Leader Peptide) 130 140 150 Patient 1 Clone αl.1/2 (SEQ ID NO: 15) TMLLVSPDIQNPDPAVYQLRDSKSSDK in Patient 2 Clone αl.3 (SEQ ID NO:17) TRLTVKPNIQNPDPAVYQLRDSKSSDK Patient 2 Clone αl.4 (SEQ ID NO:19) .RVLVKPNIQNPDPAVYQLRDSKSSDK Patient 2 Clone αl7.1 (SEQ ID NO:25) VDL NIQNPDPAVYQLRDSKSSDK Patient 3 Clone αl7.2 (SEQ ID NO:23) VDL NIQNPDPAVYQLRDSKSSDK Patient 4 Clone αl7.3 (SEQ ID NO:21) VDL NIQNPDPAVYQLRDSKSSDK Patient 4 Clone αl7.4 (SEQ ID NO:27) VDL NIQNPDPAVYQLRDSKSSDK Patient 7 Clone αl7.5 (SEQ ID NO:29) VIP NIQNPDPAVYQLRDSKSSDK Patient 4 Clone αl7.6 (SEQ ID NO:31) VKP NIQNPDPAVYQLRDSKSSDK RHEUMATOID SYNOVIAL T CELL RECEPTOR BETA CHAINS CONSERVED RESIDUES FG P POSITION (Includes Leader Peptide) 120 130 140 150 160 Patient 6 Clone 014.1 (SEQ ID NO:l) NSKT...FGS...GTRFSWEDLNKVFPPEVAVFEPSEAE Patient 6 Clone 014.2 (SEQ ID NO:3) .EAF...FGQ...GTRLTWEDLNKVFPPEVAVFEPSEAE ω Patient 5 Clone 014.3/4/5/6 (SEQ ID NO: 5) TTTDTQYFGP...GTRLTVLEDLKNVFPPEIAVFEPSEAE <τ» Patient 5 Clone 014.7 (SE ID NO:7) EQY FGP GTRLTVTEDLKNVFPPEVAVFEPSEAE Patient 5 Clone 014.8 (SEQ ID NO:9) ..TDTQYFGP...GTRLTVLEDLKNVFPPEVAVFEPSEAE Patient 6 Clone 017.1 (SEQ ID NO:11) GLTGAKNIQYFGAGTRPSVLEDLKNVFPPEVAVFEPSEAE Patient 6 Clone 017.2 (SEQ ID N0:l3) YNEQFFGP GTRLTVLEDLKNVFPPEVAVFEPSEAE Patient 6 Clone 017.3 (SEQ ID NO:33) ASYGYT FGS GTRLTWEDLKNVFPPEVAVFEPSEAE - 35 - and numbering of sequences of known T cell receptor regions. Kabat, E.A., et al., Seguences of Proteins of Immunological Interest" 4th ed. , U.S. Department of Health and Human Services, Public Health Service National Institute (1987) . Transfer and Probing Agarose gels were transferred to nylon fibers(Genescreen Plus, Du Pont New England Nuclear, Boston, MA) by capillary transfer overnight. Hybridization was with either Cα5' or C05' primers noted in Figure 5. Oligonucleotide labeling employed 100 ng DNA, 75 μC f 32P-ATP, 2.5 μl 10 x kinase buffer (500 mM Tris HCL pH 7.6, 100 mM MgCL 2 , 50 mM dithiothreitol, 1 mM sper adine, 1 mM EDTA) , 10 U T4 DNA kinase adjusted to a final volume of 25 μl with distilled water. Labelling was carried out by incubation at 37°C for 30 minutes prior to use. Blots were prehybridized in 5x SSC, 5x Denhardt's solution, 0.1% SDS for 1-1.5 hours at 55°C in sealable polyethylene bags, most of the solution poured off, 32 P-labelled oligonucleotide added (75μCi) and hybridized for 2-3 hours at 42°C or overnight at 4°C, the blots washed lx in 2x SSC, 0.1% SDS for 20 minutes at 45°C, then 3x in 5x SSC, 0.1% SDS for 20 minutes at 45°C, an exposed to Kodak XRP film at -70°C for 2-72 hours. Statistics The standard error of occurrence of each TCR V region family was calculated by the formulae: 100 times the square root of (pfl-pl/n) where "n" is the number of samples analyzed, and "p" is the number of positives. The frequency of occurrence of a particular TCR V region family was considered significantly increased if it was >.2 standard errors higher than the mean for all V regions of that type (α versus 0) . - 36 - Results PCR Primers Primers derived from the human TCR alpha and beta constant regions were utilized in conjunction with primers specific for individual variable region families. (14-16) . The primers utilized in these studies are listed in Fig. 5, and their relative positions on the coding strand of cDNA indicated in Figure 1. The constant region primers were designed as antisense primers to allow their use to prime both PCR reactions as well as probes for blotting. Variable region primers were designed to act in a family specific manner as has been previously reported (14-16) . The PCR program used in these studies employed a low stringency initial 5 cycles, followed by 40 cycles at higher stringency. The rationale for using this program was two¬ fold. As these studies were designed to investigate the range of T cell receptors expressed in RA synovium, and all TCR V regions have not yet been sequenced, related TCR families which have sequences related to the primers used here may also be amplified in the initial low stringency cycles. 40 cycles of amplification were then used to amplify even low frequency transcripts. This should help overcome the potential problem of sampling error, which is possible from surgical specimens. Thus, if local accumulations of specific TCR bearing T cells are present, and such a local accumulation is missed in the surgical specimen, their presence still may be detected if they are also present at lower frequency in the surgical specimen examined. Preliminary experiments with these primers utilizing the program described in Materials and Methods revealed that all of them (except V016) are effective in amplifying TCR V regions from PHA stimulated peripheral blood mononuclear cells, but that only the appropriate V region primers amplified Jurkat cell cDNA TCR ((20) and data not shown) . Synovial T Cells RNA was extracted and cDNA synthesized from both whole synovium and PHA stimulated/IL-2-maintained synovial T cell lines. Synovial T cell lines derived in this - 37 - manner have been previously described (17) , and early on represent a phenotypically mixed population, including CD8+ and CD4+ cells (17) . FACS analysis was available for 4 of these cell lines at the time of analysis, and the data is shown in Table 3. In 3 of these, CD4+ cells predominated, while in the other, CD8+ cells were more prevalent. TABLE 3 PHENOTYPE OF SYNOVIAL TISSUE T CELL LINES PATIENT CONTROL CD3+ CD4+ CD8+ EP, 1% NJ., 1% W, 1% HR., 3% T cell receptor transcripts were amplified from cDNA derived rom rheumatoid synovial T cell lines. All rheumatoid synovia were obtained at the time of joint surgery, and thus represented late disease. cDNA was split into equal portions and amplified with the middle constant region primers (C0 mfd or Cα mfd ) in combination with each of the respective individual variable region primers noted in Fig. 5 (eg. , C0 m] - d -i-C01, C0 mϊd +C02, ...C0 mid +C020; Cα mι - d +Cαl; Cα mid +Cα2, Cα mid + Cαl8) . Following electrophoresis and transfer, these were probed with C05' or Cα5' respectively. The results for the synovial T cell lines is shown in Figure 2. An average of 15 alpha chain and 15.8 beta chain families were detected in these cell lines. This suggests that a quite heterogeneous population of T cells is present in synovium. However, as these cell lines were initially expanded with PHA, it is possible that the proportion of the various TCR subsets alter during culture. In addition, the ability of PHA to activate resting T cells raises concern about the relative proportion of activated T cells following stimulation compared with prior to stimulation. Therefore, similar analyses were performed on whole, unstimulated rheumatoid synovium. Rheumatoid synovium The results for the whole synovia or freshly isolated, unstimulated synovial T cells analyzed similarly are - 38 - shown in Figure 3. An average of 4.2 alpha chain and 9.7 beta chain families were detected by this technique. The intensity of the bands detected is quite variable in Figure 2 & 3. To further evaluate the technique, cDNA was pooled from 4 synovia, and amplified with these primers for increasing numbers of cycles (Figure 4) . Note that the intensity of some bands which appeared in early cycles faded relative to the intensity of bands which arose at later cycles. Thus, the intensity of the bands cannot be taken as an indicator of their relative abundance. The frequency of occurrence of each TCR variable region was tabulated for synovial tissue in Figures 6 and 7. While the T cell receptor expression seen in the synovial T cell lines is quite heterogeneous, the expression in rheumatoid synovia was somewhat more limited. Specifically, Vαl7 was present 7/10 synovia, and Vαl was present in 5/10. V014 was seen in 9/10 samples, while V017 and V012 were present in 8/10 specimens and V07 was seen in 7/10. This suggests the presence of these variable regions in many rheumatoid synovia from many different patients. When analyzed statistically, the frequency of V012, 14 & 17 were >2 standard errors above the mean values for all TCR V0s detected, and Vαl7 and >2 standard errors above the mean values for all TCR Vαa detected. - 39 - References 1. Firestein, G. , and N. Zvaifler, 1987. The pathogenesis of rheumatoid arthritis. Immunology of rheumatic diseases 13:447-461. 2. Duke, O., G.S. Panayi, G. Janossy, and L. W. Poulter, 1982. An immunohistological analysis of lymphocyte subpopulations and their microenvironment in the synovial membranes of patients with rheumatoid arthritis using monoclonal antibodies. Clin Ex. Immunol 49:22-30. 3. Gao, X., E. Ball, L. Dombrausky, N. Olsen, T. Pincus, M. Kahn, and F. Wolfe, 1988. Class II human leukocyte antigen genes and T cell receptor polymorphisms in patients with rheumatoid arthritis. Am J. Med 85:14-16. 4. Goronzy, J. , C. Weyand, and C. 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Co et al., "Humanized antibodies for antiviral therapy", Proc. Natl . Acad. Sci.USA, ** : 2869-2873 (1991). - 44 - 51. Gorman, et al. "Reshaping a therapeutic CD4 antibody", Proc. Natl . Acad. Sci . USA, 88: 4181-4185 (1991). 52. Jones et al., "Resolution of hypervariabel regions in T- cell receptor 0 chains by a modified Wu-Kabat index of amino acid diversity", Proc . Natl . Acad . Sci . USA, 87: 9138-9142 (1990) . - 45 - SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: Williams, William V. Weiner, David B. (ii) TITLE OF INVENTION: T Cell Receptor-Based Therapy for Rheumatoid Arthritis (iii) NUMBER OF SEQUENCES: 79 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: Woodcock Washburn Kurtz Mackiewicz and Norris (B) STREET: One Liberty Place - 46th Floor (C) CITY: Philadelphia (D) STATE: PA (E) COUNTRY: U.S.A. (F) ZIP: 19103 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentin Release #1.0, Version #1.25 (Vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: (B) FILING DATE: (C) CLASSIFICATION: (Viii) ATTORNEY/AGENT INFORMATION: (A) NAME: Johnson, Philip S. (B) REGISTRATION NUMBER: 27,200 (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: 215-568-3100 (B) TELEFAX: 215-568-3439 (2) INFORMATION FOR SEQ ID NO:l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 237 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..237 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: GTG ACT GAT AAG GGA GAT GTT CCT GAA GGG TAC AGT GTC TCT AGA GAG Val Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Ser Arg Glu 1 5 10 15 AAG AAG GAG CGC TTC TCC CTG ATT CTG GAG TCC GCC AGC ACC AAC CAG Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 ACA TCT ATG TAC CTC TGT GCC AGC AGT TCA CAA AAA CCC AAC AGT AAA Thr Ser Met Tyr Leu Cys Ala Ser Ser Ser Gin Lys Pro Asn Ser Lys 35 40 45 ACC TTC GGT TCG GGG ACC AGG TTG TCC GTT GTA GAG GAC CTG AAC AAG Thr Phe Gly Ser Gly Thr Arg Leu Ser Val Val Glu Asp Leu Asn Lys 50 55 60 GTG TTC CCA CCC GAG GTC GCT GTG TTT GAG CCA TCA GAA GCA GAG Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 (2) INFORMATION FOR SEQ ID NO : 2 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 79 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: Val Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Ser Arg Glu 1 5 10 15 Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 Thr Ser Met Tyr Leu Cys Ala Ser Ser Ser Gin Lys Pro Asn Ser Lys 35 40 45 Thr Phe Gly Ser Gly Thr Arg Leu Ser Val Val Glu Asp Leu Asn Lys 50 55 60 Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 (2) INFORMATION FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: • (A) LENGTH: 231 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..231 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: GTG ACT GAT AAG GGA GAT GTT CCT GAA GGG TAC AGT GTC TCT AGA GAG Val Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Ser Arg Glu 1 5 10 15 AAG AAG GAG CGC TTC TCC CTG ATT CTG GAG TCC GCC AGC ACC AAC CAG Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 ACA TCT ATG TAC CTC TGT GCC AGC AGT TGG GGG ACT GAA GCT TTC TTT Thr Ser Met Tyr Leu Cys Ala Ser Ser Trp Gly Thr Glu Ala Phe Phe 35 40 45 GGA CAA GGC ACC AGA CTC ACA GTT GTA GAG GAC CTG AAC AAG GTG TTC Gly Gin Gly Thr Arg Leu Thr Val Val Glu Asp Leu Asn Lys Val Phe 50 55 60 CCA CCC GAG GTC GCT GTG TTT GAG CCA TCA GAA GCA GAG Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 (2) INFORMATION FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 77 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID N0:4: Val Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Ser Arg Glu 1 5 10 15 Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 Thr Ser Met Tyr Leu Cys Ala Ser Ser Trp Gly Thr Glu Ala Phe Phe 35 40 45 Gly Gin Gly Thr Arg Leu Thr Val Val Glu Asp Leu Asn Lys Val Phe 50 55 60 ' VO Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 ' (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 246 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..246 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: GAG ACT GAT AAG GGA GAT GTT CCT GAA GGG TAC AGT GTC TCT AGA GAG Glu Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Ser Arg Glu 1 5 10 15 AAG AAG GAG CGC TTC TCC CTG ATT CTG GAG TCC GCC AGC ACC AAC CAG Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 ACA TCT ATG TAC CTC TGT GCC AGC AGT TTG CTC CAG CGG ACC ACC ACA Thr Ser Met Tyr Leu Cys Ala Ser Ser Leu Leu Gin Arg Thr Thr Thr 35 40 45 GAT ACG CAG TAT TTT GGC CCA GGC ACC CGG CTG ACA GTG CTC GAG GAC Asp Thr Gin Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu Glu Asp 50 55 60 CTG AAA AAC GTG TTC CCA CCC GAG ATC GCT GTG TTT GAG CCA TCA GAA Leu Lys Asn Val Phe Pro Pro Glu He Ala Val Phe Glu Pro Ser Glu 65 70 75 80 GCA GAG Ala Glu (2) INFORMATION FOR SEQ ID NO:6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 82 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: Glu Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Ser Arg Glu 1 5 10 15 Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 Thr Ser Met Tyr Leu Cys Ala Ser Ser Leu Leu Gin Arg Thr Thr Thr 35 40 45 Asp Thr Gin Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu Glu Asp 50 55 60 Leu Lys Asn Val Phe Pro Pro Glu He Ala Val Phe Glu Pro Ser Glu 65 70 75 80 Ala Glu (2) INFORMATION FOR SEQ ID NO:7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 234 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..234 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: GTG ACT GAT AAG GGA GAT GTT CCT GAA GGG TAC AGT GTC TCT AGA GAG Va AAG AAG GAG CGC TTC TCC CTG ATT CTG GAG TCC GCC AGC ACC AAC CAG Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 ACA TCT ATG TAC CTC TGT GCC AGC AGC CTG GAC AGG GGC GAG CAG TAC Thr Ser Met Tyr Leu Cys Ala Ser Ser Leu Asp Arg Gly Glu Gin Tyr 35 40 45 TTC GGG CCG GGC ACC AGG CTC ACG GTC ACA GAG GAC CTG AAA AAC GTG Phe Gly Pro Gly Thr Arg Leu Thr Val Thr Glu Asp Leu Lys Asn Val i 50 55 60 ,_, f-o TTC CCA CCC GAG GTC GCT GTG TTT GAG CCA TCA GAA GCA GAG i Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 2) INFORMATION FOR SEQ ID NO:8: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 78 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: Val Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Ser Arg Glu 1 5 10 " 15 Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 Thr Ser Met Tyr Leu Cys Ala Ser Ser Leu Asp Arg Gly Glu Gin Tyr 35 40 45 Phe Gly Pro Gly Thr Arg Leu Thr Val Thr Glu Asp Leu Lys Asn Val 50 55 60 Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 (2) INFORMATION FOR SEQ ID NO:9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 240 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..240 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: GTG ACT GAT AAG GGA GAT GTT CCT GAA GGG TAC AGT GTC TCT AGA GAG Val Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Ser Arg Glu 1 5 10 15 AAG AAG GAG CGC TTC TCC CTG ATT CTG GAG TCC GCC AGC ACC AAC CAG Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 ACA TCT ATG TAC CTC TGT GCC AGC AGT TTA ACC TCC GTC ACA GAT ACG Thr Ser Met Tyr Leu Cys Ala Ser Ser Leu Thr Ser Val Thr Asp Thr 35 40 45 CAG TAT TTT GGC CCA GGC ACC CGG CTG ACA GTG CTC GAG GAC CTG AAA Gin Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu Glu Asp Leu Lys 50 55 60 AAC GTG TTC CCA CCC GAG GTC GCT GTG TTT GAG CCA TCA GAA GCA GAG Asn Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 80 (2) INFORMATION FOR SEQ ID NO:10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 80 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein ( i) SEQUENCE DESCRIPTION: SEQ ID NO: 10: Val Thr Asp Lys Gly Asp Val Pro Glu Gly Tyr Ser Val Ser Arg Glu 1 5 10 15 Lys Lys Glu Arg Phe Ser Leu He Leu Glu Ser Ala Ser Thr Asn Gin 20 25 30 Thr Ser M Gin Tyr Phe Gly Pro Gly Thr Arg Leu Thr Val Leu Glu Asp Leu Lys 50 55 60 Asn Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 80 (2) INFORMATION FOR SEQ ID NO:11: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 252 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..252 ( i) SEQUENCE DESCRIPTION: SEQ ID NO:11: TTT CAG AAA GGA GAT ATA GCT GAA GGG TAC AGC GTC TCT CGG GAG AAG Phe Gin Lys Gly Asp He Ala Glu Gly Tyr Ser Val Ser Arg Glu Lys 1 5 10 15 AAG GAA TCC TTT CCT CTC ACT GTG ACA TCG GCC CAA AAG AAC CCG ACA Lys Glu Ser Phe Pro Leu Thr Val Thr Ser Ala Gin Lys Asn Pro Thr 20 25 30 GCT TTC TAT CTC TGT GCC AGT AGT ATT GGG GGA CAA GGG CTA ACC GGG Ala Phe Tyr Leu Cys Ala Ser Ser He Gly Gly Gin Gly Leu Thr Gly 35 40 45 GCC AAA AAC ATT CAG TAC TTC GGC GCC GGG ACC CGG CCC TCA GTG CTG Ala Lys Asn He Gin Tyr Phe Gly Ala Gly Thr Arg Pro Ser Val Leu 50 55 60 GAG GAC CTG AAA AAC GTG TTC CCA CCC GAG GTC GCT GTG TTT GAG CCA Glu Asp Leu Lys Asn Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro 65 70 75 80 TCA GAA GCA GAG Ser Glu Ala Glu (2) INFORMATION FOR SEQ ID NO:12: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 84 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein ( i) SEQUENCE DESCRIPTION: SEQ ID NO: 12: Phe Gin Lys Gly Asp He Ala Glu Gly Tyr Ser Val Ser Arg Glu Lys 1 5 10 15 Lys Glu Ser Phe Pro Leu Thr Val Thr Ser Ala Gin Lys Asn Pro Thr 20 25 30 Ala Phe Tyr Leu Cys Ala Ser Ser He Gly Gly Gin Gly Leu Thr Gly 35 40 45 Ala Lys Asn He Gin Tyr Phe Gly Ala Gly Thr Arg Pro Ser Val Leu 50 55 60 Glu Asp Leu Lys Asn Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro 65 70 75 80 (2) INFORMATION FOR SEQ ID NO:13: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 237 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..237 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: TTT CAG AAA GGA GAT ATA GCT GAA GGG TAC AAA GTC TCT CGA AAA GAG Phe Gin Lys Gly Asp He Ala Glu Gly Tyr Lys Val Ser Arg Lys Glu 1 5 10 15 AAG AGG AAT TTC CCC CTG ATC CTG GAG TCG CCC AGC CCC AAC CAG ACC Lys Arg Asn Phe Pro Leu He Leu Glu Ser Pro Ser Pro Asn Gin Thr 20 25 30 TCT CTG TAC TTC TGT GCC AGC AGT TTG GGG GGA ACC TAC AAT GAG CAG Ser Leu Tyr Phe Cys Ala Ser Ser Leu Gly Gly Thr Tyr Asn Glu Gin 35 40 45 TTC TTC GGG CCA GGG ACA CGG CTC ACC GTG CTA GAG GAC CTG AAA AAC Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu Glu Asp Leu Lys Asn 50 55 60 GTG TTC CCA CCC GAG GTC GCT GTG TTT GAG CCA TCA GAA GCA GAG Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 (2) INFORMATION FOR SEQ ID NO:14: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 79 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein ( i) SEQUENCE DESCRIPTION: SEQ ID NO:14: Phe Gin Lys Gly Asp He Ala Glu Gly Tyr Lys Val Ser Arg Lys Glu 1 5 10 15 Lys Arg Asn Phe Pro Leu He Leu Glu Ser Pro Ser Pro Asn Gin Thr 20 25 30 Ser Leu Tyr Phe Cys Ala Ser Ser Leu Gly Gly Thr Tyr Asn Glu Gin 35 40 45 Phe Phe Gly Pro Gly Thr Arg Leu Thr Val Leu Glu Asp Leu Lys Asn 50 55 60 Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 (2) INFORMATION FOR SEQ ID NO:15: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 342 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..342 (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:15: CTG AGG TGC AAC TAT TCC TAT GGG GCA ACA CCT TAT CTC TTC TGG TAT Leu Arg Cys Asn Tyr Ser Tyr Gly Ala Thr Pro Tyr Leu Phe Trp Tyr 1 5 10 15 GTC CAG TCC CCC GGC CAA GGC CTC CAG CTG CCC CTG AAG TAC TTT TCA Val Gin Ser Pro Gly Gin Gly Leu Gin Leu Pro Leu Lys Tyr Phe Ser 20 25 30 GGA GAC ACT CTG GTT CAA GGC ATT AAA GGC TTT GAG GCT GAA TTT AAG Gly Asp Thr Leu Val Gin Gly He Lys Gly Phe Glu Ala Glu Phe Lys 35 40 45 AGG AGT CAA TCT TCC TTC AAT CTG AGG AAA -CCC TCT GTG CAT TGG AGT Arg Ser Gin Ser Ser Phe Asn Leu Arg Lys Pro Ser Val His Trp Ser 50 55 60 GAT GCT GCT GAG TAC TTC TGT GCT GTG GGT GCT GAT TCA GGA TAC AGC Asp Ala Ala Glu Tyr Phe Cys Ala Val Gly Ala Asp Ser Gly Tyr Ser 65 70 75 80 ACC CTC ACC TTT GGG AAG GGG ACT ATG CTT CTA GTC TCT CCA GAT ATC Thr Leu Thr Phe Gly Lys Gly Thr Met Leu Leu Val Ser Pro Asp He 85 90 95 CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC TCT AAA TCC AGT Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp Ser Lys Ser Ser 100 105 110 GAC AAG Asp Lys (2) INFORMATION FOR SEQ ID NO:16: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 114 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: Leu Arg Cys Asn Tyr Ser Tyr Gly Ala Thr Pro Tyr Leu Phe Trp Tyr 1 5 10 15 Val Gin Ser Pro Gly Gin Gly Leu Gin Leu Pro Leu Lys Tyr Phe Ser 20 25 30 Gly Asp Thr Leu Val Gin Gly He Lys Gly Phe Glu Ala Glu Phe Lys 35 40 45 Arg Ser Gin Ser Ser Phe Asn Leu Arg Lys Pro Ser Val His Trp Ser 50 55 60 Asp Ala Ala Glu Tyr Phe Cys Ala Val Gly Ala Asp Ser Gly Tyr Ser 65 70 75 80 Thr Leu Thr Phe G Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp Ser Lys Ser Ser 100 105 110 Asp Lys (2) INFORMATION FOR SEQ ID NO:17: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 336 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double I (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..336 (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: CTG AGG TGC AAC TAT TCC TAT GGG GCA ACA CCT TAT CTC TTC TGG TAT Leu Arg Cys Asn Tyr Ser Tyr Gly Ala Thr Pro Tyr Leu Phe Trp Tyr 1 5 10 15 GTC CAG TCC CCC GGC CAA GGC CTC CAG CTG CTC CTG AAG TAC TTT TCA Val Gin Ser GGA GAC ACT CTG GTT CAA GGC ATT AAA GGC TTT GAG GCT GAA TTT AAG Gly Asp Thr Leu Val Gin Gly He Lys Gly Phe Glu Ala Glu Phe Lys 35 40 45 AGG AGT CAA TCT TCC TTC AAT CTG AGG AAA CCC TCT GTG CAT TGG AGT Arg Ser Gin Ser Ser Phe Asn Leu Arg Lys Pro Ser Val His Trp Ser 50 55 60 GAT GCT GCT GAG TAC TTC TGT GCT GTG GGT CCC ACC CAC AAT GAC ATG Asp Ala Ala Glu Tyr Phe Cys Ala Val Gly Pro Thr His Asn Asp Met 65 70 75 80 CGC TTT GGA GCA GGG ACC AGA CTG ACA GTA AAA CCA AAT ATC CAG AAC Arg Phe Gly Ala Gly Thr Arg Leu Thr Val Lys Pro Asn He Gin Asn 85 90 95 CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC TCT AAA TCC AGT GAC AAG Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp Ser Lys Ser Ser Asp Lys 100 105 110 (2) INFORMATION FOR SEQ ID NO:18: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 112 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: Leu Arg.Cys Asn Tyr Ser Tyr Gly Ala Thr Pro Tyr Leu Phe Trp Tyr 1 5 10 15 Val Gin Ser Pro Gly Gin Gly Leu Gin Leu Leu Leu Lys Tyr Phe Ser 20 25 30 Gly Asp Thr Leu Val Gin Gly He Lys Gly Phe Glu Ala Glu Phe Lys 35 40 45 Arg Ser Gin Ser Ser Phe Asn Leu Arg Lys Pro Ser Val His Trp Ser 50 55 60 Asp Ala Ala Glu Tyr Phe Cys Ala Val Gly Pro Thr His Asn Asp Met 65 70 75 80 Arg Phe Gly Ala Gly Thr Arg Leu Thr Val Lys Pro Asn He Gin Asn 85 90 95 Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp Ser Lys Ser Ser Asp Lys 100 105 110 (2) INFORMATION FOR SEQ ID NO:19: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 324 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..324 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: CTG AGG TGC AAC TAT TCC TAT TCT GGG AGT CCT GAA CTC TTC TGG TAT Leu Arg Cys Asn Tyr Ser Tyr Ser Gly Ser Pro Glu Leu Phe Trp Tyr 1 5 10 15 GTC CAG TAC TCC AGA CAA CGC CTC CAG TTA CTC TTG AGA CAC ATC TCT Val Gin Tyr Ser Arg Gin Arg Leu Gin Leu Leu Leu Arg His He Ser 20 25 30 AGA GAG AGC ATC AAA GGC TTC ACT GCT GAC CTT AAC AAA GGC GAG ACA Arg Glu Ser He Lys Gly Phe Thr Ala Asp Leu Asn Lys Gly Glu Thr 35 40 45 TCT TTC CAC CTG AAG AAA CCA TTT GCT CAA GAG GAA GAC TCA GCC ATG Ser Phe His Leu Lys Lys Pro Phe Ala Gin Glu Glu Asp Ser Ala Met 50 55 60 TAT TAC TGT GCT CTA GCG CTG CAG GCA ACA AGC TTA CTT TTG GAG GAG Tyr Tyr Cys Ala Leu Ala Leu Gin Ala Thr Ser Leu Leu Leu Glu Glu 65 70 75 80 GAA CCC AGG GTG CTA GTT AAA CCA AAT ATC CAG AAC CCT GAC CCT GCC Glu Pro Arg Val Leu Val Lys Pro Asn He Gin Asn Pro Asp Pro Ala 85 90 95 GTG TAC CAG CTG AGA GAC TCT AAA TCC AGT GAC AAG Val Tyr Gin (2) INFORMATION FOR SEQ ID NO:20: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 108 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20: Leu Arg Cys Asn Tyr Ser Tyr Ser Gly Ser Pro Glu Leu Phe Trp Tyr 1 5 . 10 15 Val Gin Tyr Ser Arg Gin Arg Leu Gin Leu Leu Leu Arg His He Ser 20 25 30 Arg Glu Ser He Lys Gly Phe Thr Ala Asp Leu Asn Lys Gly Glu Thr σ» 35 40 45 ^ Ser Phe His Leu Lys Lys Pro Phe Ala Gin Glu Glu Asp Ser Ala Met 50 55 60 Tyr Tyr Cys Ala Leu Ala Leu Gin Ala Thr Ser Leu Leu Leu Glu Glu 65 70 75 80 Glu Pro Arg Val Leu Val Lys Pro Asn He Gin Asn Pro Asp Pro Ala 85 90 95 Val Tyr Gin Leu Arg Asp Ser Lys Ser Ser Asp Lys 100 105 (2) INFORMATION FOR SEQ ID NO:21: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 352 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 2..352 ( i) SEQUENCE DESCRIPTION: SEQ ID NO:21: < I C TTG TCA CTG GAT TTA GAG TCT CTC AGC TGC AGT TAC ACA GTC AGC Leu Ser Leu Asp Leu Glu Ser Leu Ser Cys Ser Tyr Thr Val Ser 1 5 10 15 GGT TTA AGA GGG CTG TTC TGG TAT AGG CAA GAT CCT GGG AAA GGC CCT Gly Leu Arg Gly Leu Phe Trp Tyr Arg Gin Asp Pro Gly Lys Gly Pro 20 25 30 GAA TTC CTC TTC ACC CTG TAT TCA GCT GGG GAA GAA AAG GAG AAA GAA Glu Phe Leu Phe Thr Leu Tyr Ser Ala Gly Glu Glu Lys Glu Lys Glu 35 40 45 AGG CTA AAA GCC ACA TTA ACA AAG AAG GAA AGC TTT CTG CAC ATC ACA Arg Leu L GCC CCT AAA CCT GAA GAC TCA GCC ACT TAT CTC TGT GCT GCG AGG CGA Ala Pro Lys Pro Glu Asp Ser Ala Thr Tyr Leu Cys Ala Ala Arg Arg 65 70 75 TCA GAT GGC CAG AAG CTG CTC TTT GCA AGG GGA ACC ATG ' TTA AAG GTG Ser Asp Gly Gin Lys Leu Leu Phe Ala Arg Gly Thr Met Leu Lys Val 80 85 90 95 GAT CTT AAT ATC CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC Asp Leu Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp 100 105 110 TCT AAA TCC AGT GAC AAG Ser Lys Ser Ser Asp Lys 115 I (2) INFORMATION FOR SEQ ID NO:22: ^ --4 (i) SEQUENCE CHARACTERISTICS: i (A) LENGTH: 117 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein ( i) SEQUENCE DESCRIPTION: SEQ ID NO:22: Leu Ser Leu Asp Leu Glu Ser Leu Ser Cys Ser Tyr Thr Val Ser Gly 1 5 10 15 Leu Arg Gly Leu Phe Trp Tyr Arg Gin Asp Pro Gly Lys Gly Pro Glu 20 25 30 Phe Leu Phe Thr Leu Tyr Ser Ala Gly Glu Glu Lys Glu Lys Glu, Arg 35 40 45 Leu Lys Ala Thr Leu Thr Lys Lys Glu Ser Phe Leu His He Thr Ala 50 55 . 60 Pro Lys Pro Glu Asp Ser Ala Thr Tyr Leu Cys Ala Ala Arg Arg Ser 65 70 75 80 Asp Gly Gin Lys Leu Leu Phe Ala Arg Gly Thr Met Leu Lys Val Asp 85 90 95 Leu Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp Ser 100 105 110 Lys Ser Ser Asp Lys 115 (2) INFORMATION FOR SEQ ID NO:23: ON 00 (i) SEQUENCE CHARACTERISTICS: ' (A) LENGTH: 352 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 2..352 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: - C TTG TCA CTG GAT TTA GAG TCT CTC AGC TGC AGT TAC ACA GTC AGC Leu Ser Leu Asp Leu Glu Ser Leu Ser Cys Ser Tyr Thr Val Ser 10 15 GGT TTA AGA GGG CTG TTC TGG TAT AGG CAA GAT CCT GGG AAA GGC CCT Gly Leu Arg Gly Leu Phe Trp Tyr Arg Gin Asp Pro Gly Lys Gly Pro 20 25 30 GAA TTC CTC TTC ACC CTG TAT TCA GCT GGG GAA GAA AAG GAG AAA GAA Glu Phe Leu Phe Thr Leu Tyr Ser Ala Gly Glu Glu Lys Glu Lys Glu 35 40 45 AGG CTA AAA GCC ACA TTA ACA AAG AAG GAA AGC TTT CTG CAC ATC ACA Arg Leu Lys Ala Thr Leu Thr Lys Lys Glu Ser Phe Leu His He Thr 50 55 60 GCC CCT AAA CCT GAA GAC TCA GCC ACT TAT CTC TGT GCT GTG AGG CGA Ala Pro Lys Pro Glu Asp Ser Ala Thr Tyr Leu Cys Ala Val Arg Arg vo 65 70 75 TCA GAT GGC CAG AAG CTG CTC TTT GCA AGG GGA ACC ATG TTA AAG GTG Ser Asp Gly Gin Lys Leu Leu Phe Ala Arg Gly Thr Met Leu Lys Val 80 85 90 95 GAT CTT AAT ATC CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC Asp Leu ' Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp 100 105 110 TCT AAA TCC AGT GAC AAG Ser Lys Ser Ser Asp Lys 115 (2) INFORMATION FOR SEQ ID NO:24: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 117 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24: Leu Ser Leu Asp Leu Glu Ser Leu Ser Cys Ser Tyr Thr Val Ser Gly 1 5 10 15 Leu Arg Gly Leu Phe Trp Tyr Arg Gin Asp Pro Gly Lys Gly Pro Glu 20 25 30 Phe Leu Phe Thr Leu Tyr Ser Ala Gly Glu Glu Lys Glu Lys Glu Arg 35 40 45 Leu Lys Ala Thr Leu Thr Lys Lys Glu Ser Phe Leu His He Thr Ala o 50 55 60 Pro Lys Pro Glu Asp Ser Ala Thr Tyr Leu Cys Ala Val Arg Arg Ser 65 70 75 80 Asp Gly Gin Lys Leu Leu Phe Ala Arg Gly Thr Met Leu Lys Val Asp 85 90 95 Leu Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp Ser 100 105 110 Lys Ser Ser Asp Lys 115 (2) INFORMATION FOR SEQ ID NO:25: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 352 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 2..352 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25: C TTG TCA CTG GAT TTA GAG TCT CTC AGC TGC AGT TAC ACA GTC AGC Leu Ser Leu Asp Leu Glu Ser Leu Ser Cys Ser Tyr Thr Val Ser I 1 5 10 15 -_j GGT TTA AGA GGG CTG TTC TGG TAT AGG CAA GAT CCT GGG AAA GGC CCT Gly Leu Arg Gly Leu Phe Trp Tyr Arg Gin Asp Pro Gly Lys Gly Pro 20 25 30 GAA TTC CTC TTC ACC CTG TAT TCA GCT GGG GAA GAA AAG GAG AAA GAA Glu Phe Leu Phe Thr Leu Tyr Ser Ala Gly Glu Glu Lys Glu Lys Glu 35 40 45 AGG CTA AAA GCC ACA TTA ACA AAG AAG GAA AGC TTT CTG CAC ATC ACA Arg Leu Lys Ala Thr Leu Thr Lys Lys Glu Ser Phe Leu His He Thr 50 55 60 GCC CCT AAA CCT GAA GAC TCA GCC ACT TAT CTC TGT GCT GTG AGG CGA Ala P TCA GAT GGC CAG AAG CTG CTC TTT GCA AGG GGA ACC ATG TTA AAG GTG Ser Asp Gly Gin Lys Leu Leu Phe Ala Arg Gly Thr Met Leu Lys Val GAT CTT AAT ATC CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC Asp Leu Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp 100 105 110 TCT AAA TCC AGT GAC AAG Ser Lys Ser Ser Asp Lys 115 (2) INFORMATION FOR SEQ ID NO:26: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 117 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26: Leu Ser Leu Asp Leu Glu Ser Leu Ser Cys Ser Tyr Thr Val Ser Gly 1 5 10 15 Leu Arg Gly Leu Phe Trp Tyr Arg Gin Asp Pro Gly Lys Gly Pro Glu 20 25 30 Phe Leu Phe Thr Leu Tyr Ser Ala Gly Glu Glu Lys Glu Lys Glu Arg 35 40 45 Leu Lys Ala Thr Leu Thr Lys Lys Glu Ser Phe Leu His He Thr Ala 50 55 60 Pro Lys Pro Glu Asp Ser Ala Thr Tyr Leu Cys Ala Val Arg Arg Ser 65 70 75 80 Asp Gly Gin Lys Leu Leu Phe Ala Arg Gly Thr Met Leu Lys Val Asp 85 90 95 Leu Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp Ser 100 105 110 Lys Ser Ser Asp Lys 115 (2) INFORMATION FOR SEQ ID NO:27: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 352 base pairs - (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 2..352 (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:27: C TTG TCA CTG GAT TTA GAG TCT CTC AGC TGC AGT TAC ACA GTC AGC Leu Ser Leu Asp Leu Glu Ser Leu Ser Cys Ser Tyr Thr Val Ser 1 5 10 15 GGT TTA AGA TGG CTG TTC TGG TAT AGG CAA GAT CCT GGG AAA GGC CCT Gly Leu Arg Trp Leu Phe Trp Tyr Arg Gin Asp Pro Gly Lys Gly Pro 20 25 30 GAA TTC CTC TTC GCC CTG TAT TCA GCT GGG GAA GAA AAG GAG AAA GAA Glu Phe Leu Phe Ala Leu Tyr Ser Ala Gly Glu Glu Lys Glu Lys Glu 35 40 45 AGG CTA AAA GCC ACA TTA ACA AAG AAG GAA AGC TTT CTG CAC ATC ACA Arg Leu Lys Ala Thr Leu Thr Lys Lys Glu Ser Phe Leu His He Thr 50 55 60 GCC CCT AAA CCT GAA GAC TCA GCC ACT TAT CTC TGT GCT GTG AGG CGA Ala Pro Lys Pro Glu Asp Ser Ala Thr Tyr Leu Cys Ala Val Arg Arg 65 70 75 TCA GAT GGC CAG AAG CTG CTC TTT GCA AGG GGA ACC ATG TTA AAG GTG -~ι Ser Asp Gly Gin Lys Leu Leu Phe Ala Arg Gly Thr Met Leu Lys Val 4 80 85 90 95 GAT CTT AAT ATC CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC Asp Leu Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp 100 105 110 TCT AAA TCC AGT GAC AAG Ser Lys Ser Ser Asp Lys 115 (2) INFORMATION FOR SEQ ID NO:28: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 117 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:28: Leu Ser Leu Asp Leu Glu Ser Leu Ser Cys Ser Tyr Thr Val Ser Gly 1 5 10 15 Leu Arg Trp Leu Phe Trp Tyr Arg Gin Asp Pro Gly Lys Gly Pro Glu 20 25 30 Phe Leu Phe Ala Leu Tyr Ser Ala Gly Glu Glu Lys Glu Lys Glu Arg 35 40 45 Leu Lys Ala Thr Leu Thr Lys Lys Glu Ser Phe Leu His He Thr Ala 50 55 60 -_J Pro Lys Pro Glu Asp Ser Ala Thr Tyr Leu Cys Ala Val Arg Arg Ser 65 70 75 80 Asp Gly Gin Lys Leu Leu Phe Ala Arg Gly Thr Met Leu Lys Val Asp 85 t 90 95 Leu Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp Ser 100 105 110 Lys Ser Ser Asp Lys 115 (2) INFORMATION FOR SEQ ID NO:29: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 343 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 2..343 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29: A GAT GTC TCC ATG AAC TGC ACT TCT TCA AGC ATA TTT AAC ACC TGG Asp Val Ser Met Asn Cys Thr Ser Ser Ser He Phe Asn Thr Trp 1 5 10 15 CTA TGG TAC AAG CAG GAC CCT GGG GAA GGT CCT GTC CTC TTG ATA GCC Leu Trp Tyr Lys Gin Asp Pro Gly Glu Gly Pro Val Leu Leu He Ala 20 25 30 TTA TAT AAG GCT GGT GAA TTG ACC TCA AAT GGA AGA CTG ACT GCT CAG Leu Tyr Lys Ala Gly Glu Leu Thr Ser Asn Gly Arg Leu Thr Ala Gin 35 40 45 TTT GGT ATA ACC AGA AAG GAC AGC TTC CTG AAT ATC TCA GCA TCC ATA Phe Gly He Thr Arg Lys Asp Ser Phe Leu Asn He Ser Ala Ser He 50 55 60 CCT AGT GAT GTA GGC ATC TAC TTC TGT GCT GGG CAG GCC CTC ACC GGT Pro Ser Asp Val Gly He Tyr Phe Cys Ala Gly Gin Ala Leu Thr Gly 65 70 75 AAC CAG TTC TAT TTT GGG ACA GGG ACA AGT TTG ACG GTC ATT CCA AAT Asn Gin Phe Tyr Phe Gly Thr Gly Thr Ser Leu Thr Val He Pro Asn ATC CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG AGA GAC TCT AAA TCC He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp Ser Lys Ser 100 105 110 AGT GAC AAG Ser Asp Lys (2) INFORMATION FOR SEQ ID NO:30: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 114 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein ' -*J (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:30: I Asp Val Ser Met Asn Cys Thr Ser Ser Ser He Phe Asn Thr Trp Leu 1 5 10 15 Trp Tyr Lys Gin Asp Pro Gly Glu Gly Pro Val Leu Leu He Ala Leu 20 25 30 Tyr Lys Ala Gly Glu Leu Thr Ser Asn Gly Arg Leu Thr Ala Gin Phe 35 40 45 Gly He Thr Arg Lys Asp Ser Phe Leu Asn He Ser Ala Ser He Pro 50 55 60 Ser Asp Val Gly He Tyr Phe Cys Ala Gly Gin Ala Leu Thr Gly Asn 65 70 75 .. 80 Gin Phe Tyr Phe Gly Thr Gly Thr Ser Leu Thr Val He Pro Asn He 85 90 95 Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu Arg Asp Ser Lys Ser Ser 100 105 110 (2) INFORMATION FOR SEQ ID NO:31: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 408 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) ~J (ix) FEATURE: 00 (A) NAME/KEY: CDS ' (B) LOCATION: 1..408 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31: CTT GTC ACT GGA TTT AGA GTC TCT CAG CTG GTG GAG CAG AGC CCT CAA Leu Val Thr Gly Phe Arg Val Ser Gin Leu Val Glu Gin Ser Pro Gin 1 5 10 15 TCT TTG ATA GTC CAG AAA GGA GGG ATT TCA ATT ATA AAC TGT GCT TAT Ser Leu He Val Gin Lys Gly Gly He Ser He He Asn Cys Ala Tyr 20 25 30 GAG AAC ACT GCG TTT GAC TAC TTT CCA TGG TAC CAA CAA TTC CCT GGG Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr Gin Gin Phe Pro Gly 35 40 45 AAA Lys AAA GAA GGA AGA TTC ACA ATC TCC TTC AAT AAA AGT GCC AAG CAG TTC Lys Glu Gly Arg Phe Thr He Ser Phe Asn Lys Ser Ala Lys Gin Phe 65 70 75 80 TCA TTG CAT ATC ATG GAT TCC CAG CCT GGA GAC TCA GCC ACC TAC TTC Ser Leu His He Met Asp Ser Gin Pro Gly Asp Ser Ala Thr Tyr Phe 85 90 95 TGT GCA GCA GAG GGA GGA AAG CTT ATC TTC GGA CAG GGA ACG GAG TTA Cys Ala Ala Glu Gly Gly Lys Leu He Phe Gly Gin Gly Thr Glu Leu ' 100 105 110 ^ TCT GTG AAA CCC AAT ATC CAG AAC CCT GAC CCT GCC GTG TAC CAG CTG ' Ser Val Lys Pro Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu 115 120 125 AGA GAC TCT AAA TCC AGT GAC AAG Arg Asp Ser Lys Ser Ser Asp Lys 130 135 (2) INFORMATION FOR SEQ ID NO:32: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 136 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32: Leu Val Thr Gly Phe Arg Val Ser Gin Leu Val Glu Gin .Ser Pro Gin 1 5 10 15 Ser Leu He Val Gin Lys Gly Gly He Ser He He Asn Cys Ala Tyr 20 25 30 Glu Asn Thr Ala Phe Asp Tyr Phe Pro Trp Tyr Gin Gin Phe Pro Gly 35 40 45 Lys Gly Pro Ala Leu Leu He Ala He Arg Pro Asp Val Ser Glu Lys 50 55 60 Lys Glu Gly Arg Phe Thr He Ser Phe Asn Lys Ser Ala Lys Gin Phe ' 65 70 75 80 ∞ Ser Leu His He Met Asp Ser Gin Pro Gly Asp Ser Ala Thr Tyr Phe ' 85 90 95 Cys Ala Ala Glu Gly Gly Lys Leu He Phe Gly Gin Gly Thr Glu Leu 100 105 110 Ser Val Lys Pro Asn He Gin Asn Pro Asp Pro Ala Val Tyr Gin Leu 115 120 125 Arg Asp Ser Lys Ser Ser Asp Lys 130 135 (2) INFORMATION FOR SEQ ID NO:33: (i) SEQUENCE CHARACTERISTICS: ' (A) LENGTH: 240 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..240 (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:33: TTT CAG AAA GGA GAT ATA GCT GAA GGG TAC AAA GTC TCT CGA AAA GAG Phe Gin Lys Gly Asp He Ala Glu Gly Tyr Lys Val Ser Arg Lys Glu ' 1 5 10 15 oo R* AAG AGG AAT TTC CCC CTG ATC CTG GAG TCG CCC AGC CCC AAC CAG ACC ' Lys Arg Asn Phe Pro Leu He Leu Glu Ser Pro Ser Pro Asn Gin Thr 20 25 30 TCT CTG TAC TTC TGT GCC AGC AGT CCG TTC TCT CGA GCA TCC TAT GGC Ser Leu Tyr Phe Cys Ala Ser Ser Pro Phe Ser Arg Ala Ser Tyr Gly 35 40 45 TAC ACC TTC GGT TCG GGG AAC AGG TTA ACC GTT GTA GAG GAC CTG AAA Tyr Thr Phe Gly Ser Gly Asn Arg Leu Thr Val Val Glu Asp Leu Lys 50 55 60 AAC GTG TTC CCA CCC GAG GTC GCT GTG TTT GAG CCA TCA GAA GCA GAG Asn Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu (2) INFORMATION FOR SEQ ID NO:34: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 80 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34: Phe Gin Lys Gly Asp He Ala Glu Gly Tyr Lys Val Ser Arg Lys Glu 1 5 10 15 Lys Arg Asn Phe Pro Leu He Leu Glu Ser Pro Ser Pro Asn Gin Thr 20 25 30 Ser Leu Tyr Phe Cys Ala Ser Ser Pro Phe Ser Arg Ala Ser Tyr Gly oo 35 40 45 N. Tyr Thr Phe Gly Ser Gly Asn Arg Leu Thr Val Val Glu Asp Leu Lys 50 55 60 Asn Val Phe Pro Pro Glu Val Ala Val Phe Glu Pro Ser Glu Ala Glu 65 70 75 80 - 83 - (2) INFORMATION FOR SEQ ID NO:35: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) ( i) SEQUENCE DESCRIPTION: SEQ ID NO:35: CTGAGGTGCA ACTACTCA (2) INFORMATION FOR SEQ ID NO:36: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:36: GTGTTCCCAG AGGGAGCCAT TGCC (2) INFORMATION FOR SEQ ID NO:37: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37: GGTGAACAGT CAACAGGGAG A (2) INFORMATION FOR SEQ ID NO:38: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38: ACAAGCATTA CTGTACTCCT A (2) INFORMATION FOR SEQ ID NO:39: - 84 - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39: GGCCCTGAAC ATTCAGGA (2) INFORMATION FOR SEQ ID NO:40: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:40: GTCACTTTCT AGCCTGCTGA (2) INFORMATION FOR SEQ ID NO:41: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41: AGGAGCCATT GTCCAGATAA A (2) INFORMATION FOR SEQ ID NO:42: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42: GGAGAGAATG TGGAGCAGCA TC (2) INFORMATION FOR SEQ ID NO:43: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs - 85 - (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43: ATCTCAGTGC TTGTGATAAT A (2) INFORMATION FOR SEQ ID NO:44: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44: ACCCAGCTGG TGGAGCAGAG CCCT (2) INFORMATION FOR SEQ ID NO:45: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) ( i) SEQUENCE DESCRIPTION: SEQ ID NO:45: AGAAAGCAAG GACCAAGTGT T (2) INFORMATION FOR SEQ ID NO: 6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6: CAGAAGGTAA CTCAAGCGCA GACT (2) INFORMATION FOR SEQ ID NO:47: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown - 86 - (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47: GCTTATGAGA ACACTGCGT (2) INFORMATION FOR SEQ ID NO:48: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48: GCAGCTTCCC TTCCAGCAAT (2) INFORMATION FOR SEQ ID NO:49: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49: AGAACCTGAC TGCCCAGGAA (2) INFORMATION FOR SEQ ID NO:50: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:50: CATCTCCATG GACTCATATG A (2) INFORMATION FOR SEQ ID NO:51: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51: - 87 - GACTATACTA ACAGCATGT (2) INFORMATION FOR SEQ ID NO:52: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52: TGTCAGGCAA TGACAAGG 18 (2) INFORMATION FOR SEQ ID NO:53: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:53: AATAGGTCGA GACACTTGTC ACTGGA (2) INFORMATION FOR SEQ ID NO:54: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54: CTTGTCACTG GATTTAGATC TCTCAGCTG (2) INFORMATION FOR SEQ ID NO:55: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55: GTACACGGCA GGGTCAGGGT TCTGGATATT (2) INFORMATION FOR SEQ ID NO:56: - 88 - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:56: AAGAGAGAGC AAAAGGAAAC ATTCTTGAAC (2) INFORMATION FOR SEQ ID NO:57: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57: GCTGCAAGGC CACATACGAG CAAGGCGTCG (2) INFORMATION FOR SEQ ID NO:58: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:58: AAAATGAAAG AAAAACCAGA TATTCCTGAG (2) INFORMATION FOR SEQ ID NO:59: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:59: CTGAGGCCAC ATATGAGAGT GGATTTGTCA (2) INFORMATION FOR SEQ ID NO:60: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown - 89 - (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60: CAGAGAAACA AAGGAAACTT CCCTGGTCGA (2) INFORMATION FOR SEQ ID NO:61: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:61: GGGTGCGGCA GATGACTCAG GGCTGCCCAA (2) INFORMATION FOR SEQ ID NO:62: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62: ATAAATGAAA GTGTGCCAAG TCGCTTCTCA (2) INFORMATION FOR SEQ ID NO:63: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) ( i) SEQUENCE DESCRIPTION: SEQ ID NO:63: AACGTTCCGA TAGATGATTC AGGGATGCCC (2) INFORMATION FOR SEQ ID NO:64: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64: - 90 - CATTATAAAT GAAACAGTTC CAAATCGCTT (2) INFORMATION FOR SEQ ID NO:65: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65: CTTATTCAGA AAGCAGAAAT AATCAATGAG (2) INFORMATION FOR SEQ ID NO:66: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66: TCCACAGAGA AGGGAGATCT TTCCTCTGAG (2) INFORMATION FOR SEQ ID NO:67: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:67: GATACTGACA AAGGAGAAGT CTCAGATGGC (2) INFORMATION FOR SEQ ID NO:68: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68: GTGACTGATA AGGGAGATGT TCCTGAAGGG (2) INFORMATION FOR SEQ ID NO:69: - 91 - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:69: GATATAAACA AAGGAGAGAT CTCTGATGGA (2) INFORMATION FOR SEQ ID NO:70: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:70: CATGATAATC TTTATCGACG TGTTATGGGA (2) INFORMATION FOR SEQ ID NO:71: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71: TTTCAGAAAG GAGATATAGC TGAAGGGTAC (2) INFORMATION FOR SEQ ID NO:72: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72: GATGAGTCAG GAATGCCAAA GGAACGATTT (2) INFORMATION FOR SEQ ID NO:73: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown - 92 - (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:73: CAAGAAACGG AGATGCACAA GAAGCGATTC (2) INFORMATION FOR SEQ ID NO:74: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74: ACCGACAGGC TGCAGGCAGG GGCCTCCAGC (2) INFORMATION FOR SEQ ID NO:75: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) ( i) SEQUENCE DESCRIPTION: SEQ ID NO:75: CCCTAGCAGG ATCTCATAGA GGATGGTGGC (2) INFORMATION FOR SEQ ID NO:76: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:76: CCCTAGCAAG ATCTCATAGA GGATGGTGGC (2) INFORMATION FOR SEQ ID NO:77: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:77: - 93 - CTCTGCTTCT GATGGCTCAA ACACAGCGAC (2) INFORMATION FOR SEQ ID NO:78: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) ( i) SEQUENCE DESCRIPTION: SEQ ID NO:78: CTCGGGTGGG AACACCTTGT TCAGGTCCTC (2) INFORMATION FOR SEQ ID NO:79: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) ( i) SEQUENCE DESCRIPTION: SEQ ID NO:79: CTCGGGTGGG AACACGTTTT TCAGGTCCTC

Claims: - 94 - Claims 1. A method of treating rheumatoid arthritis in a mammal comprising: obtaining a sample of synovium from the mammal; identifying in said sample T cell receptor variable regions; and administering to said mammal an effective amount of antibodies to at least one of said T cell receptor variable regions or antigenic fragments thereof. 2. The method of claim 1 wherein said mammal is a human. 3. The method of claim 1 wherein said sample of synovium is synovial tissue or synovial fluid. 4. A method of treating rheumatoid arthritis in a mammal comprising: administering to said mammal an effective amount of antibodies to mammalian T cell receptor variable regions selected from the group consisting of Vαl7, Vαl, V012, V014, V017 and V07 and antigenic fragments thereof. 5. The method of claim 4 wherein said antibody is specific for at least a portion of one or more peptides having amino acid sequences as set forth in Table 1. 6. The method of claim 4 wherein the mammal is human. 7. A method for immunizing a mammal to prevent the occurrence of rheumatoid arthritis comprising: administering to said mammal mammalian T cell receptor variable regions selected from the group consisting of Vαl7, Vαl, V012, V014, V017, V07 and antigenic fragments thereof. - 95 - 8. The method of claim 7 wherein the mammal is a human and the mammalian T cell receptor variable regions are human T cell receptor variable regions. 9. The method of claim 7 wherein the mammal is a human and the mammalian T cell receptor variable regions comprise at least a portion of one of the amino acid sequences set forth in Table 1. 10. A method for immunizing a mammal to treat rheumatoid arthritis comprising: administering to said mammal mammalian T cell receptor variable regions selected from the group consisting of Vαl7, Vαl, V012, V014, V017, V07 and antigenic fragments thereof. 11. The method of claim 10 wherein the mammal is a human and the mammalian T cell receptor variable regions are human T cell receptor variable regions. 12. The method of claim 10 wherein the mammal is a human and the mammalian T cell receptor variable regions comprise at least a portion of one of the amino acid sequences set forth in Table 1. 13. A kit comprising mammalian T cell receptor variable regions selected from the group consisting of Vαl7, Vαl, V 12, V 14, V017 and V07 and antigenic fragments thereof. 14. The kit of claim 13 wherein the mammalian T cell receptor variable regions are human T cell receptor variable regions. 15. The kit of claim 14 wherein the mammalian T cell receptor variable regions comprise at least a portion of one of the amino acid sequences set forth in Table 1. - 96 - 16. A kit comprising antibodies to mammalian T cell receptor variable regions selected from the group consisting of Vαl7, Vαl, V012, V014, V017 and V07 and antigenic fragments thereof. 17. The kit of claim 16 wherein the mammalian T cell receptor variable regions are human T cell receptor variable regions. 18. The kit of claim 17 wherein the variable regions comprise at least a portion of one of the amino acid sequences set forth in Table 1.

Inventors: WEINER DAVID B, WILLIAMS WILLIAM V, WEINER, DAVID, B., WILLIAMS, WILLIAM, V.
Applicants: UNIV PENNSYLVANIA, WISTAR INST, THE TRUSTEES OF THE UNIVERSITY OF PENNSYLVANIA, THE WISTAR INSTITUTE
Application year: 1992
Main classification: C07K15
IPC codes: A61K38, A61K39, A61P29, C07K14, C07K16, C12N15, C12P21, G01N33