Skip to main content
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1995 Aug;147(2):434–444.

Spontaneous apoptosis in human thymocytes.

M Tiso 1, R Gangemi 1, A Bargellesi Severi 1, S Pizzolitto 1, M Fabbi 1, A Risso 1
PMCID: PMC1869823  PMID: 7639336

Abstract

Apoptosis seems to be involved in different stages of immune cell development. In particular, experimental evidence suggests that it is a major form of cell death in the thymus. The present analysis of human thymocytes reveals that a fraction of these cells, cultured in vitro, undergoes spontaneous apoptosis. This observation is based both on molecular (DNA fragmentation) and morphological (electron microscopic) investigations of the cells. The apoptotic thymocytes are CD3- or CD3lo, CD4lo, and CD8lo and do not express Bcl-2 protein. Furthermore, thymocytes die by apoptosis when exposed to pharmacological stimuli, such as tumor necrosis factor-alpha, dexamethasone, ATP, or Ca++ ionophore. Thus the apoptotic machinery in thymocytes can be triggered by an imbalance in growth factors in the in vitro culture media and can be modulated by various biochemical signals. The process of spontaneous apoptosis is independent of mRNA or protein synthesis, as actinomycin D and cycloheximide fail to inhibit this phenomenon. Furthermore, apoptosis seems to require active oxidative phosphorylation, as it is prevented by incubation of the cells with inhibitors of the respiratory chain.

Full text

PDF
437

Images in this article

Selected References

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

  1. Arends M. J., Wyllie A. H. Apoptosis: mechanisms and roles in pathology. Int Rev Exp Pathol. 1991;32:223–254. doi: 10.1016/b978-0-12-364932-4.50010-1. [DOI] [PubMed] [Google Scholar]
  2. Barres B. A., Hart I. K., Coles H. S., Burne J. F., Voyvodic J. T., Richardson W. D., Raff M. C. Cell death and control of cell survival in the oligodendrocyte lineage. Cell. 1992 Jul 10;70(1):31–46. doi: 10.1016/0092-8674(92)90531-g. [DOI] [PubMed] [Google Scholar]
  3. Basnakian A. G., James S. J. A rapid and sensitive assay for the detection of DNA fragmentation during early phases of apoptosis. Nucleic Acids Res. 1994 Jul 11;22(13):2714–2715. doi: 10.1093/nar/22.13.2714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Caldera L. H., Leon-Ponte M., Acquatella G., Bianco N. E., Blanca I. Bone marrow and peripheral blood natural killer cell activity in lymphomas. Its response to IL-2. Clin Exp Immunol. 1992 Apr;88(1):143–148. doi: 10.1111/j.1365-2249.1992.tb03054.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clarke A. R., Purdie C. A., Harrison D. J., Morris R. G., Bird C. C., Hooper M. L., Wyllie A. H. Thymocyte apoptosis induced by p53-dependent and independent pathways. Nature. 1993 Apr 29;362(6423):849–852. doi: 10.1038/362849a0. [DOI] [PubMed] [Google Scholar]
  6. Cohen G. M., Sun X. M., Snowden R. T., Dinsdale D., Skilleter D. N. Key morphological features of apoptosis may occur in the absence of internucleosomal DNA fragmentation. Biochem J. 1992 Sep 1;286(Pt 2):331–334. doi: 10.1042/bj2860331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cohen J. J. Apoptosis. Immunol Today. 1993 Mar;14(3):126–130. doi: 10.1016/0167-5699(93)90214-6. [DOI] [PubMed] [Google Scholar]
  8. Cohen J. J., Duke R. C., Fadok V. A., Sellins K. S. Apoptosis and programmed cell death in immunity. Annu Rev Immunol. 1992;10:267–293. doi: 10.1146/annurev.iy.10.040192.001411. [DOI] [PubMed] [Google Scholar]
  9. Egerton M., Scollay R., Shortman K. Kinetics of mature T-cell development in the thymus. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2579–2582. doi: 10.1073/pnas.87.7.2579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fabbi M., Tiso M., Gangemi R. M., Favre A., Demartini P., Bargellesi-Severi A. A novel 120-kDa antigen shared by immature human thymocytes and long-term-activated T cells. Eur J Immunol. 1994 Jan;24(1):1–7. doi: 10.1002/eji.1830240102. [DOI] [PubMed] [Google Scholar]
  11. Golstein P., Ojcius D. M., Young J. D. Cell death mechanisms and the immune system. Immunol Rev. 1991 Jun;121:29–65. doi: 10.1111/j.1600-065x.1991.tb00822.x. [DOI] [PubMed] [Google Scholar]
  12. Groux H., Monte D., Plouvier B., Capron A., Ameisen J. C. CD3-mediated apoptosis of human medullary thymocytes and activated peripheral T cells: respective roles of interleukin-1, interleukin-2, interferon-gamma and accessory cells. Eur J Immunol. 1993 Jul;23(7):1623–1629. doi: 10.1002/eji.1830230734. [DOI] [PubMed] [Google Scholar]
  13. Hockenbery D. M., Oltvai Z. N., Yin X. M., Milliman C. L., Korsmeyer S. J. Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell. 1993 Oct 22;75(2):241–251. doi: 10.1016/0092-8674(93)80066-n. [DOI] [PubMed] [Google Scholar]
  14. Jacobson M. D., Burne J. F., King M. P., Miyashita T., Reed J. C., Raff M. C. Bcl-2 blocks apoptosis in cells lacking mitochondrial DNA. Nature. 1993 Jan 28;361(6410):365–369. doi: 10.1038/361365a0. [DOI] [PubMed] [Google Scholar]
  15. Jacobson M. D., Burne J. F., Raff M. C. Programmed cell death and Bcl-2 protection in the absence of a nucleus. EMBO J. 1994 Apr 15;13(8):1899–1910. doi: 10.1002/j.1460-2075.1994.tb06459.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kizaki H., Tadakuma T., Odaka C., Muramatsu J., Ishimura Y. Activation of a suicide process of thymocytes through DNA fragmentation by calcium ionophores and phorbol esters. J Immunol. 1989 Sep 15;143(6):1790–1794. [PubMed] [Google Scholar]
  17. Lowe S. W., Schmitt E. M., Smith S. W., Osborne B. A., Jacks T. p53 is required for radiation-induced apoptosis in mouse thymocytes. Nature. 1993 Apr 29;362(6423):847–849. doi: 10.1038/362847a0. [DOI] [PubMed] [Google Scholar]
  18. McConkey D. J., Hartzell P., Amador-Pérez J. F., Orrenius S., Jondal M. Calcium-dependent killing of immature thymocytes by stimulation via the CD3/T cell receptor complex. J Immunol. 1989 Sep 15;143(6):1801–1806. [PubMed] [Google Scholar]
  19. Mower D. A., Jr, Peckham D. W., Illera V. A., Fishbaugh J. K., Stunz L. L., Ashman R. F. Decreased membrane phospholipid packing and decreased cell size precede DNA cleavage in mature mouse B cell apoptosis. J Immunol. 1994 May 15;152(10):4832–4842. [PubMed] [Google Scholar]
  20. 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]
  21. Newmeyer D. D., Farschon D. M., Reed J. C. Cell-free apoptosis in Xenopus egg extracts: inhibition by Bcl-2 and requirement for an organelle fraction enriched in mitochondria. Cell. 1994 Oct 21;79(2):353–364. doi: 10.1016/0092-8674(94)90203-8. [DOI] [PubMed] [Google Scholar]
  22. Nobes C. D., Brown G. C., Olive P. N., Brand M. D. Non-ohmic proton conductance of the mitochondrial inner membrane in hepatocytes. J Biol Chem. 1990 Aug 5;265(22):12903–12909. [PubMed] [Google Scholar]
  23. Penit C. In vivo thymocyte maturation. BUdR labeling of cycling thymocytes and phenotypic analysis of their progeny support the single lineage model. J Immunol. 1986 Oct 1;137(7):2115–2121. [PubMed] [Google Scholar]
  24. Raff M. C. Social controls on cell survival and cell death. Nature. 1992 Apr 2;356(6368):397–400. doi: 10.1038/356397a0. [DOI] [PubMed] [Google Scholar]
  25. Roos D., de Boer M. Purification and cryopreservation of phagocytes from human blood. Methods Enzymol. 1986;132:225–243. doi: 10.1016/s0076-6879(86)32010-x. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. 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]
  28. Shi Y. F., Sahai B. M., Green D. R. Cyclosporin A inhibits activation-induced cell death in T-cell hybridomas and thymocytes. Nature. 1989 Jun 22;339(6226):625–626. doi: 10.1038/339625a0. [DOI] [PubMed] [Google Scholar]
  29. Skalka M., Matyásová J., Cejková M. Dna in chromatin of irradiated lymphoid tissues degrades in vivo into regular fragments. FEBS Lett. 1976 Dec 31;72(2):271–274. doi: 10.1016/0014-5793(76)80984-2. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. 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]
  32. Testi R., Phillips J. H., Lanier L. L. Constitutive expression of a phosphorylated activation antigen (Leu 23) by CD3bright human thymocytes. J Immunol. 1988 Oct 15;141(8):2557–2563. [PubMed] [Google Scholar]
  33. Tomei L. D., Shapiro J. P., Cope F. O. Apoptosis in C3H/10T1/2 mouse embryonic cells: evidence for internucleosomal DNA modification in the absence of double-strand cleavage. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):853–857. doi: 10.1073/pnas.90.3.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Veis D. J., Sentman C. L., Bach E. A., Korsmeyer S. J. Expression of the Bcl-2 protein in murine and human thymocytes and in peripheral T lymphocytes. J Immunol. 1993 Sep 1;151(5):2546–2554. [PubMed] [Google Scholar]
  35. Williams G. T., Smith C. A., Spooncer E., Dexter T. M., Taylor D. R. Haemopoietic colony stimulating factors promote cell survival by suppressing apoptosis. Nature. 1990 Jan 4;343(6253):76–79. doi: 10.1038/343076a0. [DOI] [PubMed] [Google Scholar]
  36. Wyllie A. H. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 1980 Apr 10;284(5756):555–556. doi: 10.1038/284555a0. [DOI] [PubMed] [Google Scholar]
  37. Wyllie A. H., Morris R. G. Hormone-induced cell death. Purification ad properties of thymocytes undergoing apoptosis after glucocorticoid treatment. Am J Pathol. 1982 Oct;109(1):78–87. [PMC free article] [PubMed] [Google Scholar]
  38. Yamada T., Ohyama H., Kinjo Y., Watanabe M. Evidence for the internucleosomal breakage of chromatin in rat thymocytes irradiated in vitro. Radiat Res. 1981 Mar;85(3):544–553. [PubMed] [Google Scholar]
  39. Zheng L. M., Zychlinsky A., Liu C. C., Ojcius D. M., Young J. D. Extracellular ATP as a trigger for apoptosis or programmed cell death. J Cell Biol. 1991 Jan;112(2):279–288. doi: 10.1083/jcb.112.2.279. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES