Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1986 Jun 1;163(6):1566–1582. doi: 10.1084/jem.163.6.1566

An antigen receptor-driven, interleukin 2-independent pathway for proliferation of murine cytolytic T lymphocyte clones

PMCID: PMC2188121  PMID: 3486939

Abstract

Proliferation of T lymphocytes can be induced by IL-2, either through an autocrine pathway in which the responding cell produces its own IL-2 or through an exocrine pathway in which IL-2 secreted by Th stimulates proliferation of IL-2-dependent CTL. However, proliferation of at least some CTL clones, such as CTL L3 and CTL dB45, also can be induced by stimulation of the antigen receptor in the absence of IL-2. Stimulation of these cloned CTL with T cell-depleted allogeneic spleen cells, allogeneic tumor cells, or immobilized mAb reactive with the T cell antigen receptor (TCR) induced thymidine incorporation, entry into cell cycle, and secretion of macrophage activating factor, but these stimuli did not induce the secretion of IL-2. Several observations indicated that such proliferation of cloned CTL induced by stimulation of the TCR was independent of IL-2; IL-2 could not be detected in supernatants from stimulated CTL cells. mAbs reactive with the murine IL-2-R efficiently blocked IL-2-mediated thymidine incorporation in cloned CTL and Th, but had no inhibitory effect on TCR-driven thymidine incorporation in the CTL clones. TCR-driven thymidine incorporation in cloned Th L2 cells was profoundly inhibited by these antibodies, indicating the operation of an IL-2-mediated autocrine pathway for proliferation in this cloned Th. When antibodies to the TCR were used to stimulate cloned CTL and Th, IFN-gamma mRNA was easily shown in the cloned CTL and Th. Although IL-2 mRNA could be detected in the cloned Th, it was never observed in the cloned CTL. These findings provide evidence for the existence of a TCR-mediated, IL-2-independent pathway for induction of cellular proliferation in cloned murine CTL.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Cantrell D. A., Smith K. A. Transient expression of interleukin 2 receptors. Consequences for T cell growth. J Exp Med. 1983 Dec 1;158(6):1895–1911. doi: 10.1084/jem.158.6.1895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cerottini J. C., Engers H. D., Macdonald H. R., Brunner T. Generation of cytotoxic T lymphocytes in vitro. I. Response of normal and immune mouse spleen cells in mixed leukocyte cultures. J Exp Med. 1974 Sep 1;140(3):703–717. doi: 10.1084/jem.140.3.703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Davidson W. F., Parish C. R. A procedure for removing red cells and dead cells from lymphoid cell suspensions. J Immunol Methods. 1975 Jun;7(2-3):291–300. doi: 10.1016/0022-1759(75)90026-5. [DOI] [PubMed] [Google Scholar]
  4. Farrar J. J., Mizel S. B., Fuller-Farrar J., Farrar W. L., Hilfiker M. L. Macrophage-independent activation of helper T cells. I. Production of Interleukin 2. J Immunol. 1980 Aug;125(2):793–798. [PubMed] [Google Scholar]
  5. Gillis S., Ferm M. M., Ou W., Smith K. A. T cell growth factor: parameters of production and a quantitative microassay for activity. J Immunol. 1978 Jun;120(6):2027–2032. [PubMed] [Google Scholar]
  6. Glasebrook A. L., Fitch F. W. Alloreactive cloned T cell lines. I. Interactions between cloned amplifier and cytolytic T cell lines. J Exp Med. 1980 Apr 1;151(4):876–895. doi: 10.1084/jem.151.4.876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Glasebrook A. L., Fitch F. W. T-cell lines which cooperate in generation of specific cytolytic activity. Nature. 1979 Mar 8;278(5700):171–173. doi: 10.1038/278171a0. [DOI] [PubMed] [Google Scholar]
  8. Glasebrook A. L., Quintans J., Eisenberg L., Fitch F. W. Alloreactive cloned T cell lines. II. Polyclonal stimulation of B cells by a cloned helper T cell line. J Immunol. 1981 Jan;126(1):240–244. [PubMed] [Google Scholar]
  9. Glasebrook A. L., Sarmiento M., Loken M. R., Dialynas D. P., Quintans J., Eisenberg L., Lutz C. T., Wilde D., Fitch F. W. Murine T lymphocyte clones with distinct immunological functions. Immunol Rev. 1981;54:225–266. doi: 10.1111/j.1600-065x.1981.tb00439.x. [DOI] [PubMed] [Google Scholar]
  10. Haskins K., Hannum C., White J., Roehm N., Kubo R., Kappler J., Marrack P. The antigen-specific, major histocompatibility complex-restricted receptor on T cells. VI. An antibody to a receptor allotype. J Exp Med. 1984 Aug 1;160(2):452–471. doi: 10.1084/jem.160.2.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hemler M. E., Brenner M. B., McLean J. M., Strominger J. L. Antigenic stimulation regulates the level of expression of interleukin 2 receptor on human T cells. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2172–2175. doi: 10.1073/pnas.81.7.2172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kelso A., Glasebrook A. L., Kanagawa O., Brunner K. T. Production of macrophage-activating factor by T lymphocyte clones and correlation with other lymphokine activities. J Immunol. 1982 Aug;129(2):550–556. [PubMed] [Google Scholar]
  13. Kelso A., Glasebrook A. L. Secretion of interleukin 2, macrophage-activating factor, interferon, and colony-stimulating factor by alloreactive T lymphocyte clones. J Immunol. 1984 Jun;132(6):2924–2931. [PubMed] [Google Scholar]
  14. Krishan A. Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining. J Cell Biol. 1975 Jul;66(1):188–193. doi: 10.1083/jcb.66.1.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lancki D. W., Lorber M. I., Loken M. R., Fitch F. W. A clone-specific monoclonal antibody that inhibits cytolysis of a cytolytic T cell clone. J Exp Med. 1983 Mar 1;157(3):921–935. doi: 10.1084/jem.157.3.921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lancki D. W., Ma D. I., Havran W. L., Fitch F. W. Cell surface structures involved in T cell activation. Immunol Rev. 1984 Oct;81:65–94. doi: 10.1111/j.1600-065x.1984.tb01105.x. [DOI] [PubMed] [Google Scholar]
  17. Lancki D. W., Prystowsky M. B., Vogel S. N., Beller D. I., Dialynas D. P., Fitch F. W. Effects of monoclonal antibodies directed against murine T lymphocyte cell surface antigens on lymphokine production by cloned T lymphocytes reactive with class I MHC or Mls alloantigens. J Immunol. 1984 Oct;133(4):2051–2057. [PubMed] [Google Scholar]
  18. Lowenthal J. W., Zubler R. H., Nabholz M., MacDonald H. R. Similarities between interleukin-2 receptor number and affinity on activated B and T lymphocytes. Nature. 1985 Jun 20;315(6021):669–672. doi: 10.1038/315669a0. [DOI] [PubMed] [Google Scholar]
  19. Lutz C. T., Glasebrook A. L., Fitch F. W. Alloreactive cloned T cell lines. IV. Interaction of alloantigen and T cell growth factors (TCGF) to stimulate cloned cytolytic T lymphocytes. J Immunol. 1981 Jul;127(1):391–392. [PubMed] [Google Scholar]
  20. Malek T. R., Ortega G., Jakway J. P., Chan C., Shevach E. M. The murine IL 2 receptor. II. Monoclonal anti-IL 2 receptor antibodies as specific inhibitors of T cell function in vitro. J Immunol. 1984 Oct;133(4):1976–1982. [PubMed] [Google Scholar]
  21. Malek T. R., Robb R. J., Shevach E. M. Identification and initial characterization of a rat monoclonal antibody reactive with the murine interleukin 2 receptor-ligand complex. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5694–5698. doi: 10.1073/pnas.80.18.5694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Meuer S. C., Hussey R. E., Cantrell D. A., Hodgdon J. C., Schlossman S. F., Smith K. A., Reinherz E. L. Triggering of the T3-Ti antigen-receptor complex results in clonal T-cell proliferation through an interleukin 2-dependent autocrine pathway. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1509–1513. doi: 10.1073/pnas.81.5.1509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983 Dec 16;65(1-2):55–63. doi: 10.1016/0022-1759(83)90303-4. [DOI] [PubMed] [Google Scholar]
  24. Ortega G., Robb R. J., Shevach E. M., Malek T. R. The murine IL 2 receptor. I. Monoclonal antibodies that define distinct functional epitopes on activated T cells and react with activated B cells. J Immunol. 1984 Oct;133(4):1970–1975. [PubMed] [Google Scholar]
  25. Pimlott N. J., Miller R. G. A glycopeptide extract can inhibit cytotoxic T lymphocyte-target cell conjugation in an H-2-restricted manner. J Immunol. 1984 Oct;133(4):1763–1768. [PubMed] [Google Scholar]
  26. Roopenian D. C., Widmer M. B., Orosz C. G., Bach F. H. Helper cell-independent cytolytic T lymphocytes specific for a minor histocompatibility antigen. J Immunol. 1983 Feb;130(2):542–545. [PubMed] [Google Scholar]
  27. Ryser J. E., Cerottini J. C., Brunner K. T. Generation of cytolytic T lymphocytes in vitro. IX. induction of secondary CTL responses in primary long-term MLC by supernatants from secondary MLC. J Immunol. 1978 Feb;120(2):370–377. [PubMed] [Google Scholar]
  28. Sarmiento M., Glasebrook A. L., Fitch F. W. IgG or IgM monoclonal antibodies reactive with different determinants on the molecular complex bearing Lyt 2 antigen block T cell-mediated cytolysis in the absence of complement. J Immunol. 1980 Dec;125(6):2665–2672. [PubMed] [Google Scholar]
  29. Sarmiento M., Loken M. R., Fitch F. W. Structural differences in cell surface T25 polypeptides from thymocytes and cloned T cells. Hybridoma. 1981;1(1):13–26. doi: 10.1089/hyb.1.1981.1.13. [DOI] [PubMed] [Google Scholar]
  30. Sprent J., Schaefer M. Properties of purified T cell subsets. I. In vitro responses to class I vs. class II H-2 alloantigens. J Exp Med. 1985 Dec 1;162(6):2068–2088. doi: 10.1084/jem.162.6.2068. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Staerz U. D., Rammensee H. G., Benedetto J. D., Bevan M. J. Characterization of a murine monoclonal antibody specific for an allotypic determinant on T cell antigen receptor. J Immunol. 1985 Jun;134(6):3994–4000. [PubMed] [Google Scholar]
  32. Thomas P. S. Hybridization of denatured RNA transferred or dotted nitrocellulose paper. Methods Enzymol. 1983;100:255–266. doi: 10.1016/0076-6879(83)00060-9. [DOI] [PubMed] [Google Scholar]
  33. Widmer M. B., Bach F. H. Antigen-driven helper cell-independent cloned cytolytic T lymphocytes. Nature. 1981 Dec 24;294(5843):750–752. doi: 10.1038/294750a0. [DOI] [PubMed] [Google Scholar]
  34. Yokota T., Arai N., Lee F., Rennick D., Mosmann T., Arai K. Use of a cDNA expression vector for isolation of mouse interleukin 2 cDNA clones: expression of T-cell growth-factor activity after transfection of monkey cells. Proc Natl Acad Sci U S A. 1985 Jan;82(1):68–72. doi: 10.1073/pnas.82.1.68. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES