Abstract
There are at least 63 tandemly arranged human T-cell receptor (Tcr) β-chain variable region (BV) gene segments, which have presumably arisen by repeated gene duplication events. The 5′-most half of the TCRBV gene loci is particularly complex in organization due to the presence of multiple interspersed members of the largest BV subfamilies, BV5, BV6, and BV13. Polymorphism and linkage relationships among these genes has been poorly characterized in part due to the high similarity of these duplicands. Germline DNA polymorphisms were specifically examined in the exons and introns of these and other BV gene segments distributed across 240 kilobases (kb) in this 5′-most region. Polymerase chain reaction restriction enzyme-based assays were used to genotype ten point mutations in seven of the BV gene segments. Eight of these polymorphisms altered an amino acid of the BV gene segment. In addition, length polymorphisms due to simple sequence repeats were noted in the introns of six BV6 subfamily members. Approximately 250 unrelated haplotypes were constructed by segregation analyses of fifteen of these TCRBV polymorphisms. Linkage disequilibrium analyses indicated that haplotypic relationships are not detectable over a distance of more than 55 kb in this genomic region. These TCRBV polymorphisms, and the haplotypic analysis, provide important resources and guidance for future attempts to associate Tcr germline DNA differences in the human population with immune response differences, such as might occur in some autoimmune diseases.
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References
Ball, J. K. and Desselberger, U. Incorporation of single-stranded DNA binding protein early in polymerase chain reaction product sequencing reactions prevents enzyme pausing. Anal Biochem 207: 349–351, 1992
Charmley, P. and Concannon, P. Polymorphism and phylogeny of dinucleotide repeats in human T-cell receptor Vb6 genes. Immunogenetics 38: 92–97, 1993
Charmley, P., Nepom, B. S., and Concannon, P. HLA and T cell receptor β-chain DNA polymorphisms identify a distinct subset of patients with pauciarticular-onset juvenile rheumatoid arthritis. Arthritis Rheum 37: 695–701, 1994
Charmley, P., Chao, A., Concannon, P., Hood, L., and Gatti, R. A. Haplotyping the human T-cell receptor β-chain gene complex by use of restriction fragment length polymorphisms. Proc Natl Acad Sci USA 87: 4823–4827, 1990
Charmley, P., Wang, K., Hood, L., and Nickerson, D. Identification and physical mapping of a polymorphic human T-cell receptor Vβ-gene with a frequent null allele. J Exp Med 177: 135–143, 1993
Chothia, C., Boswell, D. R., and Lesk, A.M. The outline structure of the T-cell αβ receptor. EMBO J 7: 3745–3755, 1988
Concannon, P. T-cell receptor gene polymorphisms: applications in the study of autoimmune disease. In N. R. Rose, E. C. De Macario, J. L. Fahey, H. Friedman, and G. M. Penn (Eds.): Manual of Clinical Laboratoty Immunology, 4th edn., Chapter 133, American Society for Microbiology, Washington D.C., pp. 885–889, 1992
Concannon, P., Gatti, R. A., anf Hood, L. E. Human T cell receptor Vβ gene polymorphism. J Exp Med 165: 1130–1140, 1987
Dausset, J., Cann, H., Cohen, D., Lathrop, M., Lalouel, J.-M., and White, R. Centre d'Etude du Polymorphisme Humain (CEPH): collaborative genetic mapping of the human genome. Genomics 6: 575–577, 1990
Day, C. E., Schmitt, K., and Robinson, M. A. Frequent recombination in the human T-cell receptor beta gene complex. Immunogenetics 39: 335–342, 1994
Don, R. H., Cox, P. T., Wainwright, B. J., and Mattick, J. S. ‘Touch-down’ PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res 19: 4008, 1991
Gomolka, M., Epplen, C., Buitkamp, J., and Epplen, J. T. Novel members and germline polymorphism in the human T-cell receptor Vb6 family. Immunogenetics 37: 257–265, 1993
Gomolka, M., Epplen, C., and Epplen, J. T. Germline polymorphism in the human T cell receptor Vβ6-family revealed by multiplex PCR and electroblot hybridization. Second International Conference of DNA Fingerprinting, Abstract P12, Belo Horizonte, Brazil, November 9–12, 1992
Gomolka, M., Hundrieser, J., Nurnberg, P., Roewer, L., Epplen, J. T., and Epplen, C. Selected di- and tetranucleotide microsatellites from chromosomes 7, 12, 14 and Y in various Eurasian populations. Hum Genet 93: 592–596, 1994
Hansen, T., Ronningen, K. S., Ploski, R., Kimura, A., and Thorsby, E. Coding region polymorphisms of human T-cell receptor Vβ6.9 and Vβ21.4. Scand J Immunol 36: 285–290, 1992
Hedrick, P. W., Thompson, G., and Klitz, W. In S. Karlin and E. Nevo (eds.): Evolutionary Processes and Theory, pp. 583–606, Academic Press, San Diego CA, 1986
Kretz, K. A., Carson, G. S., and O'Brien, J. S. Direct sequencing from low-melt agarose with Sequenase. Nucleic Acids Res 17: 5864, 1989
Lewontin, R. C. The interaction of selection and linkage. I. General considerations; heterotic models. Genetics 49: 49, 1964
Li, Y., Szabo, P., Robinson, M. A., Dong, B., and Posnett, D. N. Allelic variations in the human T cell receptor Vβ6.7 gene products. J Exp Med 171: 221–230, 1990
Li, Y., Szabo, P., and Posnett, D. N. The genomic structure of human Vβ6 T cell antigen receptor genes. J Exp Med 174: 1537–1547, 1991
Luyrink, L., Gabriel, C. A., Thompson, S. D., Grom, A. A., Maksymowych, W. P., Choi, E., and Glass, D. N. Reduced expression of a human Vβ6.1 T-cell receptor allele. Proc Natl Acad Sci USA 90: 4369–4373, 1993
Maksymowych, W. P., Gabriel, C. A., Luyrink, L., Melin-Aldana, H., Elma, M., Giannini, E. H., Lovell, D. J., Van Derckhove, C., Leiden, J., Choi, E., and Glass, D. N. Polymorphism in a T-cell receptor variable gene is associated with susceptibility to a juvenile rheumatoid arthritis subset. Immunogenetics 35: 257–262, 1992
Plaza, A., Kono, D. H., and Theofilopoulos, A. N. New human Vβ genes and polymorphic variants. J Immunol 147: 4360–4365, 1991
Ploski, R., Hansen, T., and Førre, Ø. Lack of association with T-cell receptor TCRBV6S1*2 allele in HLA-DQA1*0101-positive Norwegian juvenile chronic arthritis patients. Immunogenetics 38: 444–445, 1993
Posnett, D. N., Vissinga, C. S., Pambuccian, C., Wei, S., Robinson, M. A., Kostyu, D., and Concannon, P. Level of human TCRBV3S1 (Vβ3) expression correlates with allelic polymorphism in the spacer region of the recombination signal sequence. J Exp Med 179: 1707–1711, 1994
Robinson, M. A. Allelic sequence variations in the hypervariable region of a T-cell receptor β chain: correlation with restriction fragment length polymorphism in human families and populations. Proc Natl Acad Sci USA 86: 9422–9426, 1989
Vissinga, C. S., Charmley, P., and Concannon, P. Influence of coding region polymorphism on the peripheral expression of a human TCR Vβ gene. J Immunol 152: 1222–1227, 1994
Weber, J. L. Informativeness of human (dC-dA)n. (dG-dT)n polymorphisms. Genomics 7: 524–530, 1990
Wei, S., Charmley, P., Robinson, M. A., and Concannon, P. The extent of the human germline T-cell receptor V beta gene segment repertoire. Immunogenetics 40: 27–36, 1994
Wei, S., Charmley, P., and Concannon, P. Human T-cell receptor Vβ gene polymorphism and multiple sclerosis. Am J Hum Genet 56: 963–970, 1995
Zhao, T. M., Whitaker, S. E., and Robinson, M. A. A genetically determined insertion/deletion related polymorphism in human T cell receptor β chain (TCRB) includes functional variable gene segments. J Exp Med 180: 1405–1414, 1994
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Charmley, P., Concannon, P. PCR-based genotyping and haplotype analysis of human TCRBV gene segment polymorphisms. Immunogenetics 42, 254–261 (1995). https://doi.org/10.1007/BF00176442
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DOI: https://doi.org/10.1007/BF00176442