Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1990 Apr 1;171(4):1269–1281. doi: 10.1084/jem.171.4.1269

Interleukin 2 induction of pore-forming protein gene expression in human peripheral blood CD8+ T cells

PMCID: PMC2187847  PMID: 1691263

Abstract

Our studies have analyzed pore-forming protein (PFP) mRNA expression in resting and stimulated human peripheral blood CD3- large granular lymphocytes (LGL), CD3+ T cells, and their CD4+ or CD8+ subsets. Signals that stimulate T cells to develop cytotoxic activity (i.e., IL- 2 or OKT-3 mAb) led to the induction of PFP mRNA in T cells. The data indicated that IL-2 directly increased PFP mRNA in the CD8+ subset of T cells, in the absence of new DNA or protein synthesis. Abrogation of IL- 2-induced PFP mRNA expression and cytotoxic potential of T cells by the anti-p75 IL-2 receptor mAb suggested that low numbers of p75 IL-2 receptors on CD8+ T cells were capable of transducing signals responsible for these IL-2-induced effects. The induction of T cell PFP mRNA via CD3, using OKT-3 mAb, was less rapid but greater than that caused by IL-2; however, a combination of PMA and ionomycin, which bypasses crosslinking of the TCR/CD3 complex, could not mimic this increase in PFP mRNA levels in T cells. The role of second messenger systems in regulating PFP mRNA expression remains to be determined. In contrast, high constitutive PFP mRNA expression was observed in CD3- LGL and these mRNA levels could not be enhanced by stimulation with IL- 2. The cytotoxic potential of peripheral blood T cells and LGL induced in response to IL-2 correlated with IL-2-induced PFP mRNA levels in these cells and was consistent with PFP being one of several important molecules involved in the effector function of cytotoxic lymphocytes.

Full Text

The Full Text of this article is available as a PDF (1.0 MB).

Selected References

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

  1. Berrebi G., Takayama H., Sitkovsky M. V. 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. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1364–1368. doi: 10.1073/pnas.84.5.1364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  3. Ferris D. K., Willette-Brown J., Ortaldo J. R., Farrar W. L. IL-2 regulation of tyrosine kinase activity is mediated through the p70-75 beta-subunit of the IL-2 receptor. J Immunol. 1989 Aug 1;143(3):870–876. [PubMed] [Google Scholar]
  4. Garcia-Sanz J. A., Plaetinck G., Velotti F., Masson D., Tschopp J., MacDonald H. R., Nabholz M. Perforin is present only in normal activated Lyt2+ T lymphocytes and not in L3T4+ cells, but the serine protease granzyme A is made by both subsets. EMBO J. 1987 Apr;6(4):933–938. doi: 10.1002/j.1460-2075.1987.tb04841.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Granelli-Piperno A., Keane M., Steinman R. M. Growth factor production and requirements during the proliferative response of human T lymphocytes to anti-CD3 monoclonal antibody. J Immunol. 1989 Jun 15;142(12):4138–4143. [PubMed] [Google Scholar]
  6. 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]
  7. Henkart P. A., Millard P. J., Reynolds C. W., Henkart M. P. Cytolytic activity of purified cytoplasmic granules from cytotoxic rat large granular lymphocyte tumors. J Exp Med. 1984 Jul 1;160(1):75–93. doi: 10.1084/jem.160.1.75. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Heron I., Hokland M., Møller-Larsen A., Berg K. The effect of interferon on lymphocyte-mediated effector cell functions: selective enhancement of natural killer cells. Cell Immunol. 1979 Jan;42(1):183–187. doi: 10.1016/0008-8749(79)90232-6. [DOI] [PubMed] [Google Scholar]
  9. Isakov N., Mally M. I., Scholz W., Altman A. T-lymphocyte activation: the role of protein kinase C and the bifurcating inositol phospholipid signal transduction pathway. Immunol Rev. 1987 Feb;95:89–111. doi: 10.1111/j.1600-065x.1987.tb00501.x. [DOI] [PubMed] [Google Scholar]
  10. Jung G., Honsik C. J., Reisfeld R. A., Müller-Eberhard H. J. Activation of human peripheral blood mononuclear cells by anti-T3: killing of tumor target cells coated with anti-target-anti-T3 conjugates. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4479–4483. doi: 10.1073/pnas.83.12.4479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lichtenheld M. G., Olsen K. J., Lu P., Lowrey D. M., Hameed A., Hengartner H., Podack E. R. Structure and function of human perforin. Nature. 1988 Sep 29;335(6189):448–451. doi: 10.1038/335448a0. [DOI] [PubMed] [Google Scholar]
  12. Martin D. E., Zalman L. S., Jung G., Müller-Eberhard H. J. Induction of synthesis of the cytolytic C9 (ninth component of complement)-related protein in human peripheral mononuclear cells by monoclonal antibody OKT3 or interleukin 2: correlation with cytotoxicity and lymphocyte phenotype. Proc Natl Acad Sci U S A. 1987 May;84(9):2946–2950. doi: 10.1073/pnas.84.9.2946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Merćep M., Bonifacino J. S., Garcia-Morales P., Samelson L. E., Klausner R. D., Ashwell J. D. T cell CD3-zeta eta heterodimer expression and coupling to phosphoinositide hydrolysis. Science. 1988 Oct 28;242(4878):571–574. doi: 10.1126/science.2845582. [DOI] [PubMed] [Google Scholar]
  14. Millard P. J., Henkart M. P., Reynolds C. W., Henkart P. A. Purification and properties of cytoplasmic granules from cytotoxic rat LGL tumors. J Immunol. 1984 Jun;132(6):3197–3204. [PubMed] [Google Scholar]
  15. Nagler-Anderson C., Allbritton N. L., Verret C. R., Eisen H. N. A comparison of the cytolytic properties of murine primary CD8+ cytotoxic T lymphocytes and cloned cytotoxic T cell lines. Immunol Rev. 1988 Mar;103:111–125. doi: 10.1111/j.1600-065x.1988.tb00753.x. [DOI] [PubMed] [Google Scholar]
  16. Nelson D. L., Bundy B. M., Pitchon H. E., Blaese R. M., Strober W. The effector cells in human peripheral blood mediating mitogen-induced cellular cytotoxicity and antibody-dependent cellular cytotoxicity. J Immunol. 1976 Nov;117(5 Pt 1):1472–1481. [PubMed] [Google Scholar]
  17. Nishimura T., Burakoff S. J., Herrmann S. H. Protein kinase C required for cytotoxic T lymphocyte triggering. J Immunol. 1987 Nov 1;139(9):2888–2891. [PubMed] [Google Scholar]
  18. Ortaldo J. R., Longo D. L. Human natural lymphocyte effector cells: definition, analysis of activity, and clinical effectiveness. J Natl Cancer Inst. 1988 Sep 7;80(13):999–1010. doi: 10.1093/jnci/80.13.999. [DOI] [PubMed] [Google Scholar]
  19. Ortaldo J. R., Pestka S., Slease R. B., Rubinstein M., Herberman R. B. Augmentation of human K-cell activity with interferon. Scand J Immunol. 1980;12(5):365–369. doi: 10.1111/j.1365-3083.1980.tb00079.x. [DOI] [PubMed] [Google Scholar]
  20. Phillips J. H., Lanier L. L. Dissection of the lymphokine-activated killer phenomenon. Relative contribution of peripheral blood natural killer cells and T lymphocytes to cytolysis. J Exp Med. 1986 Sep 1;164(3):814–825. doi: 10.1084/jem.164.3.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Phillips J. H., Takeshita T., Sugamura K., Lanier L. L. Activation of natural killer cells via the p75 interleukin 2 receptor. J Exp Med. 1989 Jul 1;170(1):291–296. doi: 10.1084/jem.170.1.291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Saltzman E. M., Thom R. R., Casnellie J. E. Activation of a tyrosine protein kinase is an early event in the stimulation of T lymphocytes by interleukin-2. J Biol Chem. 1988 May 25;263(15):6956–6959. [PubMed] [Google Scholar]
  23. Shinkai Y., Takio K., Okumura K. Homology of perforin to the ninth component of complement (C9). Nature. 1988 Aug 11;334(6182):525–527. doi: 10.1038/334525a0. [DOI] [PubMed] [Google Scholar]
  24. Shinkai Y., Yoshida M. C., Maeda K., Kobata T., Maruyama K., Yodoi J., Yagita H., Okumura K. Molecular cloning and chromosomal assignment of a human perforin (PFP) gene. Immunogenetics. 1989;30(6):452–457. doi: 10.1007/BF02421177. [DOI] [PubMed] [Google Scholar]
  25. Takayama H., Sitkovsky M. V. Antigen receptor-regulated exocytosis in cytotoxic T lymphocytes. J Exp Med. 1987 Sep 1;166(3):725–743. doi: 10.1084/jem.166.3.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Takeshita T., Goto Y., Tada K., Nagata K., Asao H., Sugamura K. Monoclonal antibody defining a molecule possibly identical to the p75 subunit of interleukin 2 receptor. J Exp Med. 1989 Apr 1;169(4):1323–1332. doi: 10.1084/jem.169.4.1323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tanaka Y., Tozawa H., Hayami M., Sugamura K., Hinuma Y. Distinct reactivities of four monoclonal antibodies with human interleukin 2 receptor. Microbiol Immunol. 1985;29(10):959–972. doi: 10.1111/j.1348-0421.1985.tb02960.x. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Timonen T., Ortaldo J. R., Herberman R. B. Characteristics of human large granular lymphocytes and relationship to natural killer and K cells. J Exp Med. 1981 Mar 1;153(3):569–582. doi: 10.1084/jem.153.3.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Timonen T., Reynolds C. W., Ortaldo J. R., Herberman R. B. Isolation of human and rat natural killer cells. J Immunol Methods. 1982;51(3):269–277. doi: 10.1016/0022-1759(82)90393-3. [DOI] [PubMed] [Google Scholar]
  31. Trinchieri G., Matsumoto-Kobayashi M., Clark S. C., Seehra J., London L., Perussia B. Response of resting human peripheral blood natural killer cells to interleukin 2. J Exp Med. 1984 Oct 1;160(4):1147–1169. doi: 10.1084/jem.160.4.1147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yagita H., Nakata M., Azuma A., Nitta T., Takeshita T., Sugamura K., Okumura K. Activation of peripheral blood T cells via the p75 interleukin 2 receptor. J Exp Med. 1989 Oct 1;170(4):1445–1450. doi: 10.1084/jem.170.4.1445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Young H. A., Ortaldo J. R. One-signal requirement for interferon-gamma production by human large granular lymphocytes. J Immunol. 1987 Aug 1;139(3):724–727. [PubMed] [Google Scholar]
  34. Young J. D., Liu C. C., Persechini P. M., Cohn Z. A. Perforin-dependent and -independent pathways of cytotoxicity mediated by lymphocytes. Immunol Rev. 1988 Mar;103:161–202. doi: 10.1111/j.1600-065x.1988.tb00755.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES