Skip to main content
PMC home page
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1992 May;88(2):335–340. doi: 10.1111/j.1365-2249.1992.tb03083.x

T cell receptor V beta variable gene family expression in human peripheral blood lymphocytes at the mRNA and membrane protein level.

H Z Hu 1, R A de Weger 1, K Bosboom-Kalsbeek 1, M G Tilanus 1, J Rozing 1, H J Schuurman 1
PMCID: PMC1554311  PMID: 1533355

Abstract

A polymerase chain reaction (PCR) was developed, using combinations of an oligonucleotide primer for a T cell receptor V beta gene family and one for the constant C beta gene segments, to assess the expression of each of 20 V beta gene families in RNA after reverse transcription into cDNA. The detection was done after agarose gel electrophoresis of PCR products and ethidium bromide staining. The positive identification of the PCR products was done by hybridization with a J beta oligonucleotide probe. For T cell lines, a signal was observed in the V beta 8 combination for Jurkat cells, V beta 5a in HSB cells, V beta 2 and V beta 12a in Molt-3 cells and V beta 2, V beta 5a and V beta 12a in Molt-4 cells. Using mixtures of RNA from different cell lines, the sensitivity of the method was in the range of 0.1-0.5%. In peripheral blood mononuclear cells from four donors, taken at three different occasions, all V beta families were detectable. The intensity of the PCR product varied between various V beta gene families. Flow cytometric analysis of blood mononuclear cells from the same donors with a restricted series of V beta gene family-specific antibodies also revealed the presence of all families. The approach to assess V beta gene family expression in heterogeneous populations opens the possibility to study T cell receptor variable gene expression in relation to physiology and pathologic processes.

Full text

PDF
338

Images in this article

339Fig. 3
on p.339

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Acha-Orbea H., Palmer E. Mls--a retrovirus exploits the immune system. Immunol Today. 1991 Oct;12(10):356–361. doi: 10.1016/0167-5699(91)90066-3. [DOI] [PubMed] [Google Scholar]
  2. Berger M. S. Prognostic factors in lymphomas: the contribution of immunophenotyping and molecular studies. Curr Opin Immunol. 1989;2(5):719–722. doi: 10.1016/0952-7915(90)90037-h. [DOI] [PubMed] [Google Scholar]
  3. Bigler R. D., Fisher D. E., Wang C. Y., Rinnooy Kan E. A., Kunkel H. G. Idiotype-like molecules on cells of a human T cell leukemia. J Exp Med. 1983 Sep 1;158(3):1000–1005. doi: 10.1084/jem.158.3.1000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boylston A. W., Borst J., Yssel H., Blanchard D., Spits H., de Vries J. E. Properties of a panel of monoclonal antibodies which react with the human T cell antigen receptor on the leukemic line HPB-ALL and a subset of normal peripheral blood T lymphocytes. J Immunol. 1986 Jul 15;137(2):741–744. [PubMed] [Google Scholar]
  5. Choi Y. W., Kotzin B., Herron L., Callahan J., Marrack P., Kappler J. Interaction of Staphylococcus aureus toxin "superantigens" with human T cells. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8941–8945. doi: 10.1073/pnas.86.22.8941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Davies T. F., Martin A., Concepcion E. S., Graves P., Cohen L., Ben-Nun A. Evidence of limited variability of antigen receptors on intrathyroidal T cells in autoimmune thyroid disease. N Engl J Med. 1991 Jul 25;325(4):238–244. doi: 10.1056/NEJM199107253250404. [DOI] [PubMed] [Google Scholar]
  7. Ferradini L., Roman-Roman S., Azocar J., Michalaki H., Triebel F., Hercend T. Studies on the human T cell receptor alpha/beta variable region genes. II. Identification of four additional V beta subfamilies. Eur J Immunol. 1991 Apr;21(4):935–942. doi: 10.1002/eji.1830210412. [DOI] [PubMed] [Google Scholar]
  8. Kappler J., Kotzin B., Herron L., Gelfand E. W., Bigler R. D., Boylston A., Carrel S., Posnett D. N., Choi Y., Marrack P. V beta-specific stimulation of human T cells by staphylococcal toxins. Science. 1989 May 19;244(4906):811–813. doi: 10.1126/science.2524876. [DOI] [PubMed] [Google Scholar]
  9. Li Y., Szabo P., Robinson M. A., Dong B., Posnett D. N. Allelic variations in the human T cell receptor V beta 6.7 gene products. J Exp Med. 1990 Jan 1;171(1):221–230. doi: 10.1084/jem.171.1.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Maecker H. T., Levy R. Prevalence of antigen receptor variants in human T cell lines and tumors. J Immunol. 1989 Feb 15;142(4):1395–1404. [PubMed] [Google Scholar]
  11. Moller D. R., Konishi K., Kirby M., Balbi B., Crystal R. G. Bias toward use of a specific T cell receptor beta-chain variable region in a subgroup of individuals with sarcoidosis. J Clin Invest. 1988 Oct;82(4):1183–1191. doi: 10.1172/JCI113715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Oksenberg J. R., Stuart S., Begovich A. B., Bell R. B., Erlich H. A., Steinman L., Bernard C. C. Limited heterogeneity of rearranged T-cell receptor V alpha transcripts in brains of multiple sclerosis patients. Nature. 1990 May 24;345(6273):344–346. doi: 10.1038/345344a0. [DOI] [PubMed] [Google Scholar]
  13. Paliard X., West S. G., Lafferty J. A., Clements J. R., Kappler J. W., Marrack P., Kotzin B. L. Evidence for the effects of a superantigen in rheumatoid arthritis. Science. 1991 Jul 19;253(5017):325–329. doi: 10.1126/science.1857971. [DOI] [PubMed] [Google Scholar]
  14. Posnett D. N., Wang C. Y., Friedman S. M. Inherited polymorphism of the human T-cell antigen receptor detected by a monoclonal antibody. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7888–7892. doi: 10.1073/pnas.83.20.7888. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sangster R. N., Minowada J., Suciu-Foca N., Minden M., Mak T. W. Rearrangement and expression of the alpha, beta, and gamma chain T cell receptor genes in human thymic leukemia cells and functional T cells. J Exp Med. 1986 Jun 1;163(6):1491–1508. doi: 10.1084/jem.163.6.1491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sprent J., Gao E. K., Webb S. R. T cell reactivity to MHC molecules: immunity versus tolerance. Science. 1990 Jun 15;248(4961):1357–1363. doi: 10.1126/science.1694041. [DOI] [PubMed] [Google Scholar]
  17. Steinmetz M., Uematsu Y. Heterogeneity of T cell repertoires in human autoimmune disease. Br J Rheumatol. 1991;30 (Suppl 2):24–27. [PubMed] [Google Scholar]
  18. Wilson R. K., Lai E., Concannon P., Barth R. K., Hood L. E. Structure, organization and polymorphism of murine and human T-cell receptor alpha and beta chain gene families. Immunol Rev. 1988 Jan;101:149–172. doi: 10.1111/j.1600-065x.1988.tb00736.x. [DOI] [PubMed] [Google Scholar]
  19. von Boehmer H., Kisielow P. Self-nonself discrimination by T cells. Science. 1990 Jun 15;248(4961):1369–1373. doi: 10.1126/science.1972594. [DOI] [PubMed] [Google Scholar]

Articles from Clinical and Experimental Immunology are provided here courtesy of British Society for Immunology

RESOURCES