Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1993 Nov 1;178(5):1745–1751. doi: 10.1084/jem.178.5.1745

Immature thymocytes become sensitive to calcium-mediated apoptosis with the onset of CD8, CD4, and the T cell receptor expression: a role for bcl-2?

PMCID: PMC2191237  PMID: 8228820

Abstract

During intrathymic negative selection by clonal deletion, crosslinking of the T cell receptor (TCR) induces cell death by delivering an apoptotic signal(s) to the nucleus along a calcium-dependent pathway. We investigated the reactivity of early precursor-containing thymocytes to Ca(2+)-induced signals, and discovered a breakpoint in their sensitivity to calcium-mediated cell death (CMCD). CD25+CD8-4- TCR- (triple negative [TN]) thymocytes stimulated with a calcium ionophore maintain their viability and precursor activity. By contrast, their immediate progeny, CD25-CD8lo4loTCR alpha beta lo (triple low [TL]) cells react to calcium elevation by abrogation of precursor activity and apoptotic cell death. This developmental difference is specific for CMCD, since both CD25+TN and CD25-TL cells are susceptible to steroid- induced apoptosis. The presence of bcl-2 mRNA correlates directly to the resistance to CMCD-CD25+ TN cells express it and CD25-TL cells do not. These experiments show that thymocytes become sensitive to Ca(2+)- induced apoptosis as soon as they begin to express molecules that mediate thymic selection, and suggest that a concomitant downregulation of bcl-2 may mediate this phenomenon.

Full Text

The Full Text of this article is available as a PDF (740.6 KB).

Selected References

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

  1. Andjelić S., Jain N., Nikolić-Zugić J. Ontogeny of fetal CD8lo4lo thymocytes: expression of CD44, CD25 and early expression of TcR alpha mRNA. Eur J Immunol. 1993 Sep;23(9):2109–2115. doi: 10.1002/eji.1830230910. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Cohen J. J. Programmed cell death in the immune system. Adv Immunol. 1991;50:55–85. doi: 10.1016/s0065-2776(08)60822-6. [DOI] [PubMed] [Google Scholar]
  4. Finkel T. H., Cambier J. C., Kubo R. T., Born W. K., Marrack P., Kappler J. W. The thymus has two functionally distinct populations of immature alpha beta + T cells: one population is deleted by ligation of alpha beta TCR. Cell. 1989 Sep 22;58(6):1047–1054. doi: 10.1016/0092-8674(89)90503-5. [DOI] [PubMed] [Google Scholar]
  5. Finkel T. H., Kappler J. W., Marrack P. C. Immature thymocytes are protected from deletion early in ontogeny. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3372–3374. doi: 10.1073/pnas.89.8.3372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jenkins M. K., Schwartz R. H., Pardoll D. M. Effects of cyclosporine A on T cell development and clonal deletion. Science. 1988 Sep 23;241(4873):1655–1658. doi: 10.1126/science.241.4873.1655. [DOI] [PubMed] [Google Scholar]
  7. Murphy K. M., Heimberger A. B., Loh D. Y. Induction by antigen of intrathymic apoptosis of CD4+CD8+TCRlo thymocytes in vivo. Science. 1990 Dec 21;250(4988):1720–1723. doi: 10.1126/science.2125367. [DOI] [PubMed] [Google Scholar]
  8. Nicoletti I., Migliorati G., Pagliacci M. C., Grignani F., Riccardi C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991 Jun 3;139(2):271–279. doi: 10.1016/0022-1759(91)90198-o. [DOI] [PubMed] [Google Scholar]
  9. Nikolic-Zugic J., Moore M. W. T cell receptor expression on immature thymocytes with in vivo and in vitro precursor potential. Eur J Immunol. 1989 Oct;19(10):1957–1960. doi: 10.1002/eji.1830191030. [DOI] [PubMed] [Google Scholar]
  10. Nikolić-Zugić J., Bevan M. J. Thymocytes expressing CD8 differentiate into CD4+ cells following intrathymic injection. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8633–8637. doi: 10.1073/pnas.85.22.8633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Nuñez G., London L., Hockenbery D., Alexander M., McKearn J. P., Korsmeyer S. J. Deregulated Bcl-2 gene expression selectively prolongs survival of growth factor-deprived hemopoietic cell lines. J Immunol. 1990 May 1;144(9):3602–3610. [PubMed] [Google Scholar]
  13. 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]
  14. Payvar F., DeFranco D., Firestone G. L., Edgar B., Wrange O., Okret S., Gustafsson J. A., Yamamoto K. R. Sequence-specific binding of glucocorticoid receptor to MTV DNA at sites within and upstream of the transcribed region. Cell. 1983 Dec;35(2 Pt 1):381–392. doi: 10.1016/0092-8674(83)90171-x. [DOI] [PubMed] [Google Scholar]
  15. Petrie H. T., Pearse M., Scollay R., Shortman K. Development of immature thymocytes: initiation of CD3, CD4, and CD8 acquisition parallels down-regulation of the interleukin 2 receptor alpha chain. Eur J Immunol. 1990 Dec;20(12):2813–2815. doi: 10.1002/eji.1830201243. [DOI] [PubMed] [Google Scholar]
  16. Sentman C. L., Shutter J. R., Hockenbery D., Kanagawa O., Korsmeyer S. J. bcl-2 inhibits multiple forms of apoptosis but not negative selection in thymocytes. Cell. 1991 Nov 29;67(5):879–888. doi: 10.1016/0092-8674(91)90361-2. [DOI] [PubMed] [Google Scholar]
  17. Shi Y. F., Bissonnette R. P., Parfrey N., Szalay M., Kubo R. T., Green D. R. In vivo administration of monoclonal antibodies to the CD3 T cell receptor complex induces cell death (apoptosis) in immature thymocytes. J Immunol. 1991 May 15;146(10):3340–3346. [PubMed] [Google Scholar]
  18. Shi Y., Glynn J. M., Guilbert L. J., Cotter T. G., Bissonnette R. P., Green D. R. Role for c-myc in activation-induced apoptotic cell death in T cell hybridomas. Science. 1992 Jul 10;257(5067):212–214. doi: 10.1126/science.1378649. [DOI] [PubMed] [Google Scholar]
  19. Smith C. A., Williams G. T., Kingston R., Jenkinson E. J., Owen J. J. Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures. Nature. 1989 Jan 12;337(6203):181–184. doi: 10.1038/337181a0. [DOI] [PubMed] [Google Scholar]
  20. Strasser A., Harris A. W., Cory S. bcl-2 transgene inhibits T cell death and perturbs thymic self-censorship. Cell. 1991 Nov 29;67(5):889–899. doi: 10.1016/0092-8674(91)90362-3. [DOI] [PubMed] [Google Scholar]
  21. Takahama Y., Singer A. Post-transcriptional regulation of early T cell development by T cell receptor signals. Science. 1992 Nov 27;258(5087):1456–1462. doi: 10.1126/science.1439838. [DOI] [PubMed] [Google Scholar]
  22. Thornthwaite J. T., Sugarbaker E. V., Temple W. J. Preparation of tissues for DNA flow cytometric analysis. Cytometry. 1980 Nov;1(3):229–237. doi: 10.1002/cyto.990010309. [DOI] [PubMed] [Google Scholar]
  23. Ucker D. S., Meyers J., Obermiller P. S. Activation-driven T cell death. II. Quantitative differences alone distinguish stimuli triggering nontransformed T cell proliferation or death. J Immunol. 1992 Sep 1;149(5):1583–1592. [PubMed] [Google Scholar]
  24. Vasquez N. J., Kaye J., Hedrick S. M. In vivo and in vitro clonal deletion of double-positive thymocytes. J Exp Med. 1992 May 1;175(5):1307–1316. doi: 10.1084/jem.175.5.1307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Watanabe-Fukunaga R., Brannan C. I., Copeland N. G., Jenkins N. A., Nagata S. Lymphoproliferation disorder in mice explained by defects in Fas antigen that mediates apoptosis. Nature. 1992 Mar 26;356(6367):314–317. doi: 10.1038/356314a0. [DOI] [PubMed] [Google Scholar]
  26. Wilson A., Petrie H. T., Scollay R., Shortman K. The acquisition of CD4 and CD8 during the differentiation of early thymocytes in short-term culture. Int Immunol. 1989;1(6):605–612. doi: 10.1093/intimm/1.6.605. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES