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. 1987 Mar;84(5):1364–1368. doi: 10.1073/pnas.84.5.1364

Antigen-receptor interaction requirement for conjugate formation and lethal-hit triggering by cytotoxic T lymphocytes can be bypassed by protein kinase C activators and Ca2+ ionophores.

G Berrebi, H Takayama, M V Sitkovsky
PMCID: PMC304430  PMID: 3103133

Abstract

We show that phorbol esters and Ca2+ ionophores can trigger the lysis of nonantigen-bearing target cells by cytotoxic T lymphocytes. This effect obviates the requirement for antigen-receptor-mediated recognition of the antigen; the intensity of lysis is dose and Ca2+ dependent and requires contact between cytotoxic T lymphocytes and target cells. Using a fluorescence-activated cell sorter to enumerate cytotoxic T lymphocyte-target cell conjugates, we show that phorbol esters at concentrations that triggered lysis of non-antigen-bearing target cells also increased the number of stable conjugates with these target cells. The results point to the importance of the antigen-nonspecific engagements of cytotoxic T lymphocytes in immunologic surveillance. The data also show that the linkage between the T-cell receptor and antigen is not mandatory for conjugate formation, for the strengthening of conjugates, and for lysis.

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Selected References

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

  1. Albert F., Hua C., Truneh A., Pierres M., Schmitt-Verhulst A. M. Distinction between antigen receptor and IL 2 receptor triggering events in the activation of alloreactive T cell clones with calcium ionophore and phorbol ester. J Immunol. 1985 Jun;134(6):3649–3655. [PubMed] [Google Scholar]
  2. Berke G. Enumeration of lymphocyte-target cell conjugates by cytofluorometry. Eur J Immunol. 1985 Apr;15(4):337–340. doi: 10.1002/eji.1830150406. [DOI] [PubMed] [Google Scholar]
  3. Farrar W. L., Anderson W. B. Interleukin-2 stimulates association of protein kinase C with plasma membrane. Nature. 1985 May 16;315(6016):233–235. doi: 10.1038/315233a0. [DOI] [PubMed] [Google Scholar]
  4. Geiger B., Rosen D., Berke G. Spatial relationships of microtubule-organizing centers and the contact area of cytotoxic T lymphocytes and target cells. J Cell Biol. 1982 Oct;95(1):137–143. doi: 10.1083/jcb.95.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Griffith B. P., Shaw B. W., Jr, Hardesty R. L., Iwatsuki S., Bahnson H. T., Starzl T. E. Veno-venous bypass without systemic anticoagulation for transplantation of the human liver. Surg Gynecol Obstet. 1985 Mar;160(3):270–272. [PMC free article] [PubMed] [Google Scholar]
  7. Henkart P. A. Mechanism of lymphocyte-mediated cytotoxicity. Annu Rev Immunol. 1985;3:31–58. doi: 10.1146/annurev.iy.03.040185.000335. [DOI] [PubMed] [Google Scholar]
  8. Horwitz A., Duggan K., Buck C., Beckerle M. C., Burridge K. Interaction of plasma membrane fibronectin receptor with talin--a transmembrane linkage. Nature. 1986 Apr 10;320(6062):531–533. doi: 10.1038/320531a0. [DOI] [PubMed] [Google Scholar]
  9. Hubbard S. C., Kranz D. M., Longmore G. D., Sitkovsky M. V., Eisen H. N. Glycosylation of the T-cell antigen-specific receptor and its potential role in lectin-mediated cytotoxicity. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1852–1856. doi: 10.1073/pnas.83.6.1852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kikkawa U., Takai Y., Tanaka Y., Miyake R., Nishizuka Y. Protein kinase C as a possible receptor protein of tumor-promoting phorbol esters. J Biol Chem. 1983 Oct 10;258(19):11442–11445. [PubMed] [Google Scholar]
  11. Kranz D. M., Tonegawa S., Eisen H. N. Attachment of an anti-receptor antibody to non-target cells renders them susceptible to lysis by a clone of cytotoxic T lymphocytes. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7922–7926. doi: 10.1073/pnas.81.24.7922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kupfer A., Singer S. J., Dennert G. On the mechanism of unidirectional killing in mixtures of two cytotoxic T lymphocytes. Unidirectional polarization of cytoplasmic organelles and the membrane-associated cytoskeleton in the effector cell. J Exp Med. 1986 Mar 1;163(3):489–498. doi: 10.1084/jem.163.3.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lanzavecchia A. Is the T-cell receptor involved in T-cell killing? 1986 Feb 27-Mar 5Nature. 319(6056):778–780. doi: 10.1038/319778a0. [DOI] [PubMed] [Google Scholar]
  14. Manger B., Weiss A., Weyand C., Goronzy J., Stobo J. D. T cell activation: differences in the signals required for IL 2 production by nonactivated and activated T cells. J Immunol. 1985 Dec;135(6):3669–3673. [PubMed] [Google Scholar]
  15. Meuer S. C., Fitzgerald K. A., Hussey R. E., Hodgdon J. C., Schlossman S. F., Reinherz E. L. Clonotypic structures involved in antigen-specific human T cell function. Relationship to the T3 molecular complex. J Exp Med. 1983 Feb 1;157(2):705–719. doi: 10.1084/jem.157.2.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. doi: 10.1038/308693a0. [DOI] [PubMed] [Google Scholar]
  17. Perez P., Bluestone J. A., Stephany D. A., Segal D. M. Quantitative measurements of the specificity and kinetics of conjugate formation between cloned cytotoxic T lymphocytes and splenic target cells by dual parameter flow cytometry. J Immunol. 1985 Jan;134(1):478–485. [PubMed] [Google Scholar]
  18. Perez P., Hoffman R. W., Shaw S., Bluestone J. A., Segal D. M. Specific targeting of cytotoxic T cells by anti-T3 linked to anti-target cell antibody. Nature. 1985 Jul 25;316(6026):354–356. doi: 10.1038/316354a0. [DOI] [PubMed] [Google Scholar]
  19. Rothlein R., Springer T. A. The requirement for lymphocyte function-associated antigen 1 in homotypic leukocyte adhesion stimulated by phorbol ester. J Exp Med. 1986 May 1;163(5):1132–1149. doi: 10.1084/jem.163.5.1132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Russell J. H. Phorbol-ester stimulated lysis of weak and nonspecific target cells by cytotoxic T lymphocytes. J Immunol. 1986 Jan;136(1):23–27. [PubMed] [Google Scholar]
  21. Sarmiento M., Dialynas D. P., Lancki D. W., Wall K. A., Lorber M. I., Loken M. R., Fitch F. W. Cloned T lymphocytes and monoclonal antibodies as probes for cell surface molecules active in T cell-mediated cytolysis. Immunol Rev. 1982;68:135–169. doi: 10.1111/j.1600-065x.1982.tb01063.x. [DOI] [PubMed] [Google Scholar]
  22. Schrezenmeier H., Kurrle R., Wagner H., Fleischer B. Activation of human T lymphocytes. III. Triggering of bystander cytotoxicity in cytotoxic T cell clones by antibodies against the T3 antigen or by a calcium ionophore. Eur J Immunol. 1985 Oct;15(10):1019–1024. doi: 10.1002/eji.1830151011. [DOI] [PubMed] [Google Scholar]
  23. Shaw S., Luce G. E., Quinones R., Gress R. E., Springer T. A., Sanders M. E. Two antigen-independent adhesion pathways used by human cytotoxic T-cell clones. Nature. 1986 Sep 18;323(6085):262–264. doi: 10.1038/323262a0. [DOI] [PubMed] [Google Scholar]
  24. Shortman K., Golstein P. Target cell recognition by cytolytic T cells: different requirements for the formation of strong conjugates or for proceeding to lysis. J Immunol. 1979 Aug;123(2):833–839. [PubMed] [Google Scholar]
  25. Siliciano R. F., Pratt J. C., Schmidt R. E., Ritz J., Reinherz E. L. Activation of cytolytic T lymphocyte and natural killer cell function through the T11 sheep erythrocyte binding protein. Nature. 1985 Oct 3;317(6036):428–430. doi: 10.1038/317428a0. [DOI] [PubMed] [Google Scholar]
  26. Sitkovsky M. V., Pasternack M. S., Lugo J. P., Klein J. R., Eisen H. N. Isolation and partial characterization of concanavalin A receptors on cloned cytotoxic T lymphocytes. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1519–1523. doi: 10.1073/pnas.81.5.1519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sitkovsky M. V., Schwartz M. A., Eisen H. N. Cell-cell contact proteins in antigen-specific and antigen-nonspecific cellular cytotoxicity. Adv Exp Med Biol. 1985;184:429–449. doi: 10.1007/978-1-4684-8326-0_29. [DOI] [PubMed] [Google Scholar]
  28. Spits H., Borst J., Terhorst C., de Vries J. E. The role of T cell differentiation markers in antigen-specific and lectin-dependent cellular cytotoxicity mediated by T8+ and T4+ human cytotoxic T cell clones directed at class I and class II MHC antigens. J Immunol. 1982 Oct;129(4):1563–1569. [PubMed] [Google Scholar]
  29. Spits H., van Schooten W., Keizer H., van Seventer G., van de Rijn M., Terhorst C., de Vries J. E. Alloantigen recognition is preceded by nonspecific adhesion of cytotoxic T cells and target cells. Science. 1986 Apr 18;232(4748):403–405. doi: 10.1126/science.3485822. [DOI] [PubMed] [Google Scholar]
  30. Staerz U. D., Kanagawa O., Bevan M. J. Hybrid antibodies can target sites for attack by T cells. Nature. 1985 Apr 18;314(6012):628–631. doi: 10.1038/314628a0. [DOI] [PubMed] [Google Scholar]
  31. Staerz U. D., Pasternack M. S., Klein J. R., Benedetto J. D., Bevan M. J. Monoclonal antibodies specific for a murine cytotoxic T-lymphocyte clone. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1799–1803. doi: 10.1073/pnas.81.6.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Truneh A., Albert F., Golstein P., Schmitt-Verhulst A. M. Early steps of lymphocyte activation bypassed by synergy between calcium ionophores and phorbol ester. Nature. 1985 Jan 24;313(6000):318–320. doi: 10.1038/313318a0. [DOI] [PubMed] [Google Scholar]
  33. Utsunomiya N., Tsuboi M., Nakanishi M. Early transmembrane events in alloimmune cytotoxic T-lymphocyte activation as revealed by stopped-flow fluorometry. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1877–1880. doi: 10.1073/pnas.83.6.1877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Weiss M. J., Daley J. F., Hodgdon J. C., Reinherz E. L. Calcium dependency of antigen-specific (T3-Ti) and alternative (T11) pathways of human T-cell activation. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6836–6840. doi: 10.1073/pnas.81.21.6836. [DOI] [PMC free article] [PubMed] [Google Scholar]

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