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. 1994 Nov;83(3):469–475.

Effects of irradiation, glucocorticoid and FK506 on cell-surface antigen expression by rat thymocytes: a three-colour flow cytofluorometric analysis.

M Tsuchida 1, M Konishi 1, K Takai 1, K Naito 1, Y Fujikura 1, T Fukumoto 1
PMCID: PMC1415031  PMID: 7530693

Abstract

The expression of T-cell antigen receptor (TCR) alpha beta was investigated in rat CD4- CD8- thymocytes during thymic reconstitution after the exposure of animals to irradiation or glucocorticoid. The effect of the immunosuppressant FK506 on the expression of TCR alpha beta in rat CD4- CD8- thymocytes was also examined. The percentage of CD4- CD8- thymocytes constituted 2.6% of total thymocytes and that of CD4- CD8- TCR alpha beta high cells constituted 12.6% of CD4- CD8- thymocytes in normal adult Lewis rats. The percentage of CD4- CD8- TCR alpha beta high cells increased during thymic reconstitution after irradiation, and maximally constituted 28.6% of CD4- CD8- thymocytes on day 7. Similar results were obtained during thymic reconstitution after glucocorticoid treatment. In contrast, continuous treatment with FK506 for 7 days markedly decreased not only the percentages of CD4+ CD8- TCR alpha beta high and CD4- CD8+ TCR alpha beta high thymocytes, but also that of CD4- CD8- TCR alpha beta high thymocytes. These results indicate that rat CD4- CD8- thymocytes contain a subpopulation of mature (TCR alpha beta high) cells. The possible implications of the existence of this subpopulation with regard to thymocyte differentiation and maturation are discussed.

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Selected References

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

  1. Blackman M., Kappler J., Marrack P. The role of the T cell receptor in positive and negative selection of developing T cells. Science. 1990 Jun 15;248(4961):1335–1341. doi: 10.1126/science.1972592. [DOI] [PubMed] [Google Scholar]
  2. Boyd R. L., Hugo P. Towards an integrated view of thymopoiesis. Immunol Today. 1991 Feb;12(2):71–79. doi: 10.1016/0167-5699(91)90161-L. [DOI] [PubMed] [Google Scholar]
  3. Brideau R. J., Carter P. B., McMaster W. R., Mason D. W., Williams A. F. Two subsets of rat T lymphocytes defined with monoclonal antibodies. Eur J Immunol. 1980 Aug;10(8):609–615. doi: 10.1002/eji.1830100807. [DOI] [PubMed] [Google Scholar]
  4. Budd R. C., Miescher G. C., Howe R. C., Lees R. K., Bron C., MacDonald H. R. Developmentally regulated expression of T cell receptor beta chain variable domains in immature thymocytes. J Exp Med. 1987 Aug 1;166(2):577–582. doi: 10.1084/jem.166.2.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clark S. J., Law D. A., Paterson D. J., Puklavec M., Williams A. F. Activation of rat T lymphocytes by anti-CD2 monoclonal antibodies. J Exp Med. 1988 Jun 1;167(6):1861–1872. doi: 10.1084/jem.167.6.1861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clipstone N. A., Crabtree G. R. Identification of calcineurin as a key signalling enzyme in T-lymphocyte activation. Nature. 1992 Jun 25;357(6380):695–697. doi: 10.1038/357695a0. [DOI] [PubMed] [Google Scholar]
  7. Crispe I. N., Moore M. W., Husmann L. A., Smith L., Bevan M. J., Shimonkevitz R. P. Differentiation potential of subsets of CD4-8- thymocytes. Nature. 1987 Sep 24;329(6137):336–339. doi: 10.1038/329336a0. [DOI] [PubMed] [Google Scholar]
  8. Denning S. M., Tuck D. T., Vollger L. W., Springer T. A., Singer K. H., Haynes B. F. Monoclonal antibodies to CD2 and lymphocyte function-associated antigen 3 inhibit human thymic epithelial cell-dependent mature thymocyte activation. J Immunol. 1987 Oct 15;139(8):2573–2578. [PubMed] [Google Scholar]
  9. Fowlkes B. J., Kruisbeek A. M., Ton-That H., Weston M. A., Coligan J. E., Schwartz R. H., Pardoll D. M. A novel population of T-cell receptor alpha beta-bearing thymocytes which predominantly expresses a single V beta gene family. Nature. 1987 Sep 17;329(6136):251–254. doi: 10.1038/329251a0. [DOI] [PubMed] [Google Scholar]
  10. Godfrey D. I., Zlotnik A. Control points in early T-cell development. Immunol Today. 1993 Nov;14(11):547–553. doi: 10.1016/0167-5699(93)90186-O. [DOI] [PubMed] [Google Scholar]
  11. Gotlieb W. H., Durum S. K., Gregorio T. A., Mathieson B. J. Selective stimulation of thymocyte precursors mediated by specific cytokines. Different CD3+ subsets are generated by IL-1 versus IL-2. J Immunol. 1991 Apr 1;146(7):2262–2271. [PubMed] [Google Scholar]
  12. Hugo P., Boyd R. L., Waanders G. A., Scollay R. CD4+CD8+CD3high thymocytes appear transiently during ontogeny: evidence from phenotypic and functional studies. Eur J Immunol. 1991 Nov;21(11):2655–2660. doi: 10.1002/eji.1830211103. [DOI] [PubMed] [Google Scholar]
  13. Hugo P., Waanders G. A., Scollay R., Shortman K., Boyd R. L. Ontogeny of a novel CD4+CD8-CD3- thymocyte subpopulation: a comparison with CD4- CD8+ CD3- thymocytes. Int Immunol. 1990;2(3):209–218. doi: 10.1093/intimm/2.3.209. [DOI] [PubMed] [Google Scholar]
  14. Hünig T., Mitnacht R. T cell receptor-mediated selection of functional rat CD8 T cells from defined immature thymocyte precursors in short-term suspension culture. J Exp Med. 1991 Mar 1;173(3):561–568. doi: 10.1084/jem.173.3.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hünig T., Wallny H. J., Hartley J. K., Lawetzky A., Tiefenthaler G. A monoclonal antibody to a constant determinant of the rat T cell antigen receptor that induces T cell activation. Differential reactivity with subsets of immature and mature T lymphocytes. J Exp Med. 1989 Jan 1;169(1):73–86. doi: 10.1084/jem.169.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jefferies W. A., Green J. R., Williams A. F. Authentic T helper CD4 (W3/25) antigen on rat peritoneal macrophages. J Exp Med. 1985 Jul 1;162(1):117–127. doi: 10.1084/jem.162.1.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kato K., Koyanagi M., Okada H., Takanashi T., Wong Y. W., Williams A. F., Okumura K., Yagita H. CD48 is a counter-receptor for mouse CD2 and is involved in T cell activation. J Exp Med. 1992 Nov 1;176(5):1241–1249. doi: 10.1084/jem.176.5.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Liu J., Farmer J. D., Jr, Lane W. S., Friedman J., Weissman I., Schreiber S. L. Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes. Cell. 1991 Aug 23;66(4):807–815. doi: 10.1016/0092-8674(91)90124-h. [DOI] [PubMed] [Google Scholar]
  19. MacDonald H. R., Budd R. C., Howe R. C. A CD3- subset of CD4-8+ thymocytes: a rapidly cycling intermediate in the generation of CD4+8+ cells. Eur J Immunol. 1988 Apr;18(4):519–523. doi: 10.1002/eji.1830180405. [DOI] [PubMed] [Google Scholar]
  20. Nikolić-Zugić J., Moore M. W., Bevan M. J. Characterization of the subset of immature thymocytes which can undergo rapid in vitro differentiation. Eur J Immunol. 1989 Apr;19(4):649–653. doi: 10.1002/eji.1830190412. [DOI] [PubMed] [Google Scholar]
  21. Nikolić-Zugić J. Phenotypic and functional stages in the intrathymic development of alpha beta T cells. Immunol Today. 1991 Feb;12(2):65–70. doi: 10.1016/0167-5699(91)90160-u. [DOI] [PubMed] [Google Scholar]
  22. O'Keefe S. J., Tamura J., Kincaid R. L., Tocci M. J., O'Neill E. A. FK-506- and CsA-sensitive activation of the interleukin-2 promoter by calcineurin. Nature. 1992 Jun 25;357(6380):692–694. doi: 10.1038/357692a0. [DOI] [PubMed] [Google Scholar]
  23. Pearse M., Wu L., Egerton M., Wilson A., Shortman K., Scollay R. A murine early thymocyte developmental sequence is marked by transient expression of the interleukin 2 receptor. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1614–1618. doi: 10.1073/pnas.86.5.1614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pieters R. H., Bol M., Lam B. W., Seinen W., Bloksma N., Penninks A. H. Recovery from chemically induced thymus atrophy starts with CD4- CD8- CD2high TcR alpha beta-/low thymocytes and results in an increased formation of CD4- CD8- TcR alpha beta high thymocytes. Immunology. 1993 Apr;78(4):616–622. [PMC free article] [PubMed] [Google Scholar]
  25. Pugh-Humphreys R. G., Ross C. S., Thomson A. W. The influence of FK-506 on the thymus: an immunophenotypic and structural analysis. Immunology. 1990 Jul;70(3):398–404. [PMC free article] [PubMed] [Google Scholar]
  26. Ramsdell F., Fowlkes B. J. Clonal deletion versus clonal anergy: the role of the thymus in inducing self tolerance. Science. 1990 Jun 15;248(4961):1342–1348. doi: 10.1126/science.1972593. [DOI] [PubMed] [Google Scholar]
  27. Reimann J., Bellan A., Conradt P. Development of autoreactive L3T4+ T cells from double-negative (L3T4-/Ly-2-) Thy-1+ spleen cells of normal mice. Eur J Immunol. 1988 Jul;18(7):989–999. doi: 10.1002/eji.1830180704. [DOI] [PubMed] [Google Scholar]
  28. Reimann J. Double-negative (CD4-CD8-), TCR alpha beta-expressing, peripheral T cells. Scand J Immunol. 1991 Dec;34(6):679–688. doi: 10.1111/j.1365-3083.1991.tb01592.x. [DOI] [PubMed] [Google Scholar]
  29. Scollay R., Bartlett P., Shortman K. T cell development in the adult murine thymus: changes in the expression of the surface antigens Ly2, L3T4 and B2A2 during development from early precursor cells to emigrants. Immunol Rev. 1984 Dec;82:79–103. doi: 10.1111/j.1600-065x.1984.tb01118.x. [DOI] [PubMed] [Google Scholar]
  30. Shimonkevitz R. P., Husmann L. A., Bevan M. J., Crispe I. N. Transient expression of IL-2 receptor precedes the differentiation of immature thymocytes. Nature. 1987 Sep 10;329(6135):157–159. doi: 10.1038/329157a0. [DOI] [PubMed] [Google Scholar]
  31. Shortman K., Wilson A., Egerton M., Pearse M., Scollay R. Immature CD4- CD8+ murine thymocytes. Cell Immunol. 1988 May;113(2):462–479. doi: 10.1016/0008-8749(88)90042-1. [DOI] [PubMed] [Google Scholar]
  32. Singer P. A., Theofilopoulos A. N. T-cell receptor V beta repertoire expression in murine models of SLE. Immunol Rev. 1990 Dec;118:103–127. doi: 10.1111/j.1600-065x.1990.tb00814.x. [DOI] [PubMed] [Google Scholar]
  33. Takahama Y., Kosugi A., Singer A. Phenotype, ontogeny, and repertoire of CD4-CD8- T cell receptor alpha beta + thymocytes. Variable influence of self-antigens on T cell receptor V beta usage. J Immunol. 1991 Feb 15;146(4):1134–1141. [PubMed] [Google Scholar]
  34. Takai K., Tokuda N., Sawada T., Fujikura Y., Jojima K., Sakatoku J., Fukumoto T. Effects of FK506 on rat thymic epithelial cells; immunohistochemical study. Thymus. 1992 Jun;19(4):207–217. [PubMed] [Google Scholar]
  35. Tanaka M., Shinohara K., Fukumoto T., Tanaka H., Kaneko T. Effect of Cyclosporin A on rat thymus: time course analysis by immunoperoxidase technique and flow cytofluorometry. Clin Exp Immunol. 1988 May;72(2):216–221. [PMC free article] [PubMed] [Google Scholar]
  36. Teh H. S., Kishi H., Scott B., Von Boehmer H. Deletion of autospecific T cells in T cell receptor (TCR) transgenic mice spares cells with normal TCR levels and low levels of CD8 molecules. J Exp Med. 1989 Mar 1;169(3):795–806. doi: 10.1084/jem.169.3.795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tiefenthaler G., Hanke T., Hünig T. Maturation of CD4-8- alpha/beta TCR+ T cells induced by CD2-stimulation in vivo and in vitro. Int Immunol. 1992 Jul;4(7):825–829. doi: 10.1093/intimm/4.7.825. [DOI] [PubMed] [Google Scholar]
  38. Williams A. F., Galfrè G., Milstein C. Analysis of cell surfaces by xenogeneic myeloma-hybrid antibodies: differentiation antigens of rat lymphocytes. Cell. 1977 Nov;12(3):663–673. doi: 10.1016/0092-8674(77)90266-5. [DOI] [PubMed] [Google Scholar]
  39. de Talance A., Regnier D., Spinella S., Morisset J., Seman M. Origin of autoreactive T helper cells. I. Characterization of Thy-1+, Lyt-, L3T4- precursors in the spleen of normal mice. J Immunol. 1986 Aug 15;137(4):1101–1108. [PubMed] [Google Scholar]
  40. von Boehmer H. Thymic selection: a matter of life and death. Immunol Today. 1992 Nov;13(11):454–458. doi: 10.1016/0167-5699(92)90075-I. [DOI] [PubMed] [Google Scholar]

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