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. 1993 May;92(2):361–366. doi: 10.1111/j.1365-2249.1993.tb03405.x

The human T cell receptor V beta repertoire of normal peripheral blood lymphocytes before and after mitogen stimulation.

F S Wong 1, M L Hibberd 1, L Wen 1, B A Millward 1, A G Demaine 1
PMCID: PMC1554814  PMID: 8387412

Abstract

Mitogen stimulation of T cells in vitro has been employed in the analysis of the T cell antigen receptor (TCR) repertoire and as a method of generating T cell lines and clones. It has been suspected for some time that mitogen stimulation may bias the repertoire. We have addressed this problem employing a semi-quantitative technique utilizing the polymerase chain reaction (PCR) and flow cytometry. Using this PCR method and a panel of primers to 22 V beta subgroups, the V beta repertoire of both unstimulated and phytohaemagglutinin (PHA)-stimulated peripheral T cells from eight healthy individuals was investigated. The samples were also analysed by flow cytometry using anti-V beta 2, V beta 5 and V beta 8 MoAbs. A significant increase in the expression of V beta 6, V beta 7.2 and V beta 10.1 was found in all eight samples of PHA-stimulated T cells compared with unstimulated T cells using the PCR method. In contrast, no differences were found between unstimulated and PHA-stimulated T cells by flow cytometry. These results question the validity of using mitogen-stimulated T cells to investigate TCR gene usage.

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Selected References

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  1. Babbitt B. P., Allen P. M., Matsueda G., Haber E., Unanue E. R. Binding of immunogenic peptides to Ia histocompatibility molecules. 1985 Sep 26-Oct 2Nature. 317(6035):359–361. doi: 10.1038/317359a0. [DOI] [PubMed] [Google Scholar]
  2. Baccala R., Kono D. H., Walker S., Balderas R. S., Theofilopoulos A. N. Genomically imposed and somatically modified human thymocyte V beta gene repertoires. Proc Natl Acad Sci U S A. 1991 Apr 1;88(7):2908–2912. doi: 10.1073/pnas.88.7.2908. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boyer C., Auphan N., Luton F., Malburet J. M., Barad M., Bizozzero J. P., Reggio H., Schmitt-Verhulst A. M. T cell receptor/CD3 complex internalization following activation of a cytolytic T cell clone: evidence for a protein kinase C-independent staurosporine-sensitive step. Eur J Immunol. 1991 Jul;21(7):1623–1634. doi: 10.1002/eji.1830210707. [DOI] [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. Cantrell D. A., Davies A. A., Crumpton M. J. Activators of protein kinase C down-regulate and phosphorylate the T3/T-cell antigen receptor complex of human T lymphocytes. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8158–8162. doi: 10.1073/pnas.82.23.8158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cossarizza A., Kahan M., Ortolani C., Franceschi C., Londei M. Preferential expression of V beta 6.7 domain on human peripheral CD4+ T cells. Implication for positive selection of T cells in man. Eur J Immunol. 1991 Jun;21(6):1571–1574. doi: 10.1002/eji.1830210637. [DOI] [PubMed] [Google Scholar]
  7. Dalgleish A. G., Wilson S., Gompels M., Ludlam C., Gazzard B., Coates A. M., Habeshaw J. T-cell receptor variable gene products and early HIV-1 infection. Lancet. 1992 Apr 4;339(8797):824–828. doi: 10.1016/0140-6736(92)90277-a. [DOI] [PubMed] [Google Scholar]
  8. Davey M. P., Meyer M. M., Munkirs D. D., Babcock D., Braun M. P., Hayden J. B., Bakke A. C. T-cell receptor variable beta genes show differential expression in CD4 and CD8 T cells. Hum Immunol. 1991 Nov;32(3):194–202. doi: 10.1016/0198-8859(91)90056-f. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Davies T. F., Martin A., Concepcion E. S., Graves P., Lahat N., Cohen W. L., Ben-Nun A. Evidence for selective accumulation of intrathyroidal T lymphocytes in human autoimmune thyroid disease based on T cell receptor V gene usage. J Clin Invest. 1992 Jan;89(1):157–162. doi: 10.1172/JCI115556. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Davis M. M. T cell receptor gene diversity and selection. Annu Rev Biochem. 1990;59:475–496. doi: 10.1146/annurev.bi.59.070190.002355. [DOI] [PubMed] [Google Scholar]
  12. Goldman A. S., Palkowetz K. H., Rudloff H. E., Brooks E. G., Schmalstieg F. C. Repertoire of V alpha and V beta regions of T cell antigen receptors on CD4+ and CD8+ peripheral blood T cells in a novel X-linked combined immunodeficiency disease. Eur J Immunol. 1992 Apr;22(4):1103–1106. doi: 10.1002/eji.1830220435. [DOI] [PubMed] [Google Scholar]
  13. Grunewald J., Janson C. H., Wigzell H. Biased expression of individual T cell receptor V gene segments in CD4+ and CD8+ human peripheral blood T lymphocytes. Eur J Immunol. 1991 Mar;21(3):819–822. doi: 10.1002/eji.1830210342. [DOI] [PubMed] [Google Scholar]
  14. Gulwani-Akolkar B., Posnett D. N., Janson C. H., Grunewald J., Wigzell H., Akolkar P., Gregersen P. K., Silver J. T cell receptor V-segment frequencies in peripheral blood T cells correlate with human leukocyte antigen type. J Exp Med. 1991 Nov 1;174(5):1139–1146. doi: 10.1084/jem.174.5.1139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Herman A., Kappler J. W., Marrack P., Pullen A. M. Superantigens: mechanism of T-cell stimulation and role in immune responses. Annu Rev Immunol. 1991;9:745–772. doi: 10.1146/annurev.iy.09.040191.003525. [DOI] [PubMed] [Google Scholar]
  16. Hu H. Z., de Weger R. A., Bosboom-Kalsbeek K., Tilanus M. G., Rozing J., Schuurman H. J. T cell receptor V beta variable gene family expression in human peripheral blood lymphocytes at the mRNA and membrane protein level. Clin Exp Immunol. 1992 May;88(2):335–340. doi: 10.1111/j.1365-2249.1992.tb03083.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Imberti L., Sottini A., Spagnoli G., Primi D. Expression of the human V beta 8 gene product preferentially correlates with class II major histocompatibility complex restriction specificity. Eur J Immunol. 1990 Dec;20(12):2817–2819. doi: 10.1002/eji.1830201244. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Lindsten T., June C. H., Thompson C. B. Transcription of T cell antigen receptor genes is induced by protein kinase C activation. J Immunol. 1988 Sep 1;141(5):1769–1774. [PubMed] [Google Scholar]
  20. Loveridge J. A., Rosenberg W. M., Kirkwood T. B., Bell J. I. The genetic contribution to human T-cell receptor repertoire. Immunology. 1991 Oct;74(2):246–250. [PMC free article] [PubMed] [Google Scholar]
  21. Marguerie C., Lunardi C., So A. PCR-based analysis of the TCR repertoire in human autoimmune diseases. Immunol Today. 1992 Sep;13(9):336–338. doi: 10.1016/0167-5699(92)90166-5. [DOI] [PubMed] [Google Scholar]
  22. Moretta A., Pantaleo G., Moretta L., Cerottini J. C., Mingari M. C. Direct demonstration of the clonogenic potential of every human peripheral blood T cell. Clonal analysis of HLA-DR expression and cytolytic activity. J Exp Med. 1983 Feb 1;157(2):743–754. doi: 10.1084/jem.157.2.743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Rosenberg W. M., Moss P. A., Bell J. I. Variation in human T cell receptor V beta and J beta repertoire: analysis using anchor polymerase chain reaction. Eur J Immunol. 1992 Feb;22(2):541–549. doi: 10.1002/eji.1830220237. [DOI] [PubMed] [Google Scholar]

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