Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1994 Dec 20;91(26):12659–12663. doi: 10.1073/pnas.91.26.12659

Interleukin 12 induces the differentiation of major histocompatibility complex class I-primed cytotoxic T-lymphocyte precursors into allospecific cytotoxic effectors.

S Chouaib 1, J Chehimi 1, L Bani 1, N Genetet 1, T Tursz 1, F Gay 1, G Trinchieri 1, F Mami-Chouaib 1
PMCID: PMC45498  PMID: 7809096

Abstract

The production of interleukin 12 (IL-12) following allogeneic stimulation and its involvement in the differentiation of allospecific cytotoxic T lymphocytes (CTLs) have been investigated. Supernatants of mixed lymphocyte cultures had detectable levels of IL-12 p40 which were completely abrogated after depletion of responder cells from monocytes. While addition to the culture of anti-IL-12 neutralizing antibodies partially inhibited the allogeneic proliferative response and the subsequent CTL activity, addition of IL-12 stimulated both responses, suggesting that endogenously produced IL-12 plays a role in the development of alloreactivity. Furthermore, using primary mixed cultures of lymphocytes from major histocompatibility complex-recombinant siblings identical for class II antigens and displaying class I disparity, we demonstrated that addition of recombinant IL-12 at the sensitizing phase of the primary mixed lymphocyte culture induced CTL activity. Under these stimulation conditions, addition of recombinant IL-12 also triggered cell proliferation, indicating that IL-12 provides both growth and differentiation signals. The mechanism underlying this process does not appear to require IL-2, since IL-12-mediated CTL generation was not abrogated by anti-IL-2 alpha-chain antibodies. IL-12 increased granzyme B and perforin mRNA accumulation in major histocompatibility complex class I-primed lymphocytes, suggesting that this cytokine activates these two genes in CTL precursors. We conclude that IL-12 can stimulate the generation of alloreactive CTLs. We suggest that IL-12 may play a role in helper cell-independent CTL generation.

Full text

PDF
12659

Images in this article

12662Fig. 6
on p.12662

Selected References

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

  1. Aste-Amezaga M., D'Andrea A., Kubin M., Trinchieri G. Cooperation of natural killer cell stimulatory factor/interleukin-12 with other stimuli in the induction of cytokines and cytotoxic cell-associated molecules in human T and NK cells. Cell Immunol. 1994 Jul;156(2):480–492. doi: 10.1006/cimm.1994.1192. [DOI] [PubMed] [Google Scholar]
  2. Bloom E. T., Horvath J. A. Cellular and molecular mechanisms of the IL-12-induced increase in allospecific murine cytolytic T cell activity. Implications for the age-related decline in CTL. J Immunol. 1994 May 1;152(9):4242–4254. [PubMed] [Google Scholar]
  3. Brunda M. J., Luistro L., Warrier R. R., Wright R. B., Hubbard B. R., Murphy M., Wolf S. F., Gately M. K. Antitumor and antimetastatic activity of interleukin 12 against murine tumors. J Exp Med. 1993 Oct 1;178(4):1223–1230. doi: 10.1084/jem.178.4.1223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Buyse I., Decorte R., Baens M., Cuppens H., Semana G., Emonds M. P., Marynen P., Cassiman J. J. Rapid DNA typing of class II HLA antigens using the polymerase chain reaction and reverse dot blot hybridization. Tissue Antigens. 1993 Jan;41(1):1–14. doi: 10.1111/j.1399-0039.1993.tb01970.x. [DOI] [PubMed] [Google Scholar]
  5. Chan S. H., Perussia B., Gupta J. W., Kobayashi M., Pospísil M., Young H. A., Wolf S. F., Young D., Clark S. C., Trinchieri G. Induction of interferon gamma production by natural killer cell stimulatory factor: characterization of the responder cells and synergy with other inducers. J Exp Med. 1991 Apr 1;173(4):869–879. doi: 10.1084/jem.173.4.869. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chehimi J., Starr S. E., Frank I., D'Andrea A., Ma X., MacGregor R. R., Sennelier J., Trinchieri G. Impaired interleukin 12 production in human immunodeficiency virus-infected patients. J Exp Med. 1994 Apr 1;179(4):1361–1366. doi: 10.1084/jem.179.4.1361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chehimi J., Starr S. E., Frank I., Rengaraju M., Jackson S. J., Llanes C., Kobayashi M., Perussia B., Young D., Nickbarg E. Natural killer (NK) cell stimulatory factor increases the cytotoxic activity of NK cells from both healthy donors and human immunodeficiency virus-infected patients. J Exp Med. 1992 Mar 1;175(3):789–796. doi: 10.1084/jem.175.3.789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chehimi J., Valiante N. M., D'Andrea A., Rengaraju M., Rosado Z., Kobayashi M., Perussia B., Wolf S. F., Starr S. E., Trinchieri G. Enhancing effect of natural killer cell stimulatory factor (NKSF/interleukin-12) on cell-mediated cytotoxicity against tumor-derived and virus-infected cells. Eur J Immunol. 1993 Aug;23(8):1826–1830. doi: 10.1002/eji.1830230814. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Chouaib S., Bensussan A., Termijtelen A. M., Andreeff M., Marchiol-Fournigault C., Fradelizi D., Dupont B. Allogeneic T cell activation triggering by MHC class I antigens. J Immunol. 1988 Jul 15;141(2):423–429. [PubMed] [Google Scholar]
  11. Chouaib S., Welte K., Dupont B. Differential effect of anti-beta 2-microglobulin on IL 2 production and IL 2 receptor expression in the primary mixed lymphocyte culture reaction. J Immunol. 1985 Feb;134(2):940–948. [PubMed] [Google Scholar]
  12. Clerici M., Lucey D. R., Berzofsky J. A., Pinto L. A., Wynn T. A., Blatt S. P., Dolan M. J., Hendrix C. W., Wolf S. F., Shearer G. M. Restoration of HIV-specific cell-mediated immune responses by interleukin-12 in vitro. Science. 1993 Dec 10;262(5140):1721–1724. doi: 10.1126/science.7903123. [DOI] [PubMed] [Google Scholar]
  13. D'Andrea A., Rengaraju M., Valiante N. M., Chehimi J., Kubin M., Aste M., Chan S. H., Kobayashi M., Young D., Nickbarg E. Production of natural killer cell stimulatory factor (interleukin 12) by peripheral blood mononuclear cells. J Exp Med. 1992 Nov 1;176(5):1387–1398. doi: 10.1084/jem.176.5.1387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dasgupta J. D., Cemach K., Dubey D. P., Yunis E. J., Amos D. B. The role of class I histocompatibility antigens in the regulation of T-cell activation. Proc Natl Acad Sci U S A. 1987 Feb;84(4):1094–1098. doi: 10.1073/pnas.84.4.1094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Finke J. H., Scott J., Gillis S., Hilfiker M. L. Generation of alloreactive cytotoxic T lymphocytes: evidence for a differentiation factor distinct from IL 2. J Immunol. 1983 Feb;130(2):763–767. [PubMed] [Google Scholar]
  16. Garman R. D., Fan D. P. Characterization of helper factors distinct from interleukin 2 necessary for the generation of allospecific cytolytic T lymphocytes. J Immunol. 1983 Feb;130(2):756–762. [PubMed] [Google Scholar]
  17. Gately M. K., Wolitzky A. G., Quinn P. M., Chizzonite R. Regulation of human cytolytic lymphocyte responses by interleukin-12. Cell Immunol. 1992 Aug;143(1):127–142. doi: 10.1016/0008-8749(92)90011-d. [DOI] [PubMed] [Google Scholar]
  18. Gubler U., Chua A. O., Schoenhaut D. S., Dwyer C. M., McComas W., Motyka R., Nabavi N., Wolitzky A. G., Quinn P. M., Familletti P. C. Coexpression of two distinct genes is required to generate secreted bioactive cytotoxic lymphocyte maturation factor. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4143–4147. doi: 10.1073/pnas.88.10.4143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kobayashi M., Fitz L., Ryan M., Hewick R. M., Clark S. C., Chan S., Loudon R., Sherman F., Perussia B., Trinchieri G. Identification and purification of natural killer cell stimulatory factor (NKSF), a cytokine with multiple biologic effects on human lymphocytes. J Exp Med. 1989 Sep 1;170(3):827–845. doi: 10.1084/jem.170.3.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Landolfo S., Cofano F., Giovarelli M., Prat M., Cavallo G., Forni G. Inhibition of interferon-gamma may suppress allograft reactivity by T lymphocytes in vitro and in vivo. Science. 1985 Jul 12;229(4709):176–179. doi: 10.1126/science.3160110. [DOI] [PubMed] [Google Scholar]
  21. Mami-Chouaib F., Miossec C., Del Porto P., Flament C., Triebel F., Hercend T. T cell target 1 (TCT.1): a novel target molecule for human non-major histocompatibility complex-restricted T lymphocytes. J Exp Med. 1990 Oct 1;172(4):1071–1082. doi: 10.1084/jem.172.4.1071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Manetti R., Parronchi P., Giudizi M. G., Piccinni M. P., Maggi E., Trinchieri G., Romagnani S. Natural killer cell stimulatory factor (interleukin 12 [IL-12]) induces T helper type 1 (Th1)-specific immune responses and inhibits the development of IL-4-producing Th cells. J Exp Med. 1993 Apr 1;177(4):1199–1204. doi: 10.1084/jem.177.4.1199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mehrotra P. T., Wu D., Crim J. A., Mostowski H. S., Siegel J. P. Effects of IL-12 on the generation of cytotoxic activity in human CD8+ T lymphocytes. J Immunol. 1993 Sep 1;151(5):2444–2452. [PubMed] [Google Scholar]
  24. Moretta A., Accolla R. S., Cerottini J. C. IL-2-mediated T cell proliferation in humans is blocked by a monoclonal antibody directed against monomorphic determinants of HLA-DR antigens. J Exp Med. 1982 Feb 1;155(2):599–604. doi: 10.1084/jem.155.2.599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Naume B., Gately M. K., Desai B. B., Sundan A., Espevik T. Synergistic effects of interleukin 4 and interleukin 12 on NK cell proliferation. Cytokine. 1993 Jan;5(1):38–46. doi: 10.1016/1043-4666(93)90022-w. [DOI] [PubMed] [Google Scholar]
  26. Naume B., Gately M., Espevik T. A comparative study of IL-12 (cytotoxic lymphocyte maturation factor)-, IL-2-, and IL-7-induced effects on immunomagnetically purified CD56+ NK cells. J Immunol. 1992 Apr 15;148(8):2429–2436. [PubMed] [Google Scholar]
  27. Naume B., Johnsen A. C., Espevik T., Sundan A. Gene expression and secretion of cytokines and cytokine receptors from highly purified CD56+ natural killer cells stimulated with interleukin-2, interleukin-7 and interleukin-12. Eur J Immunol. 1993 Aug;23(8):1831–1838. doi: 10.1002/eji.1830230815. [DOI] [PubMed] [Google Scholar]
  28. Perussia B., Chan S. H., D'Andrea A., Tsuji K., Santoli D., Pospisil M., Young D., Wolf S. F., Trinchieri G. Natural killer (NK) cell stimulatory factor or IL-12 has differential effects on the proliferation of TCR-alpha beta+, TCR-gamma delta+ T lymphocytes, and NK cells. J Immunol. 1992 Dec 1;149(11):3495–3502. [PubMed] [Google Scholar]
  29. Robertson M. J., Soiffer R. J., Wolf S. F., Manley T. J., Donahue C., Young D., Herrmann S. H., Ritz J. Response of human natural killer (NK) cells to NK cell stimulatory factor (NKSF): cytolytic activity and proliferation of NK cells are differentially regulated by NKSF. J Exp Med. 1992 Mar 1;175(3):779–788. doi: 10.1084/jem.175.3.779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Robinet E., Branellec D., Termijtelen A. M., Blay J. Y., Gay F., Chouaib S. Evidence for tumor necrosis factor-alpha involvement in the optimal induction of class I allospecific cytotoxic T cells. J Immunol. 1990 Jun 15;144(12):4555–4561. [PubMed] [Google Scholar]
  31. Salcedo T. W., Azzoni L., Wolf S. F., Perussia B. Modulation of perforin and granzyme messenger RNA expression in human natural killer cells. J Immunol. 1993 Sep 1;151(5):2511–2520. [PubMed] [Google Scholar]
  32. Sterkers G., Henin Y., Kalil J., Bagot M., Levy J. P. Influence of HLA class I- and class II-specific monoclonal antibodies on DR-restricted lymphoproliferative responses. I. Unseparated populations of effector cells. J Immunol. 1983 Dec;131(6):2735–2740. [PubMed] [Google Scholar]
  33. Stern A. S., Podlaski F. J., Hulmes J. D., Pan Y. C., Quinn P. M., Wolitzky A. G., Familletti P. C., Stremlo D. L., Truitt T., Chizzonite R. Purification to homogeneity and partial characterization of cytotoxic lymphocyte maturation factor from human B-lymphoblastoid cells. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6808–6812. doi: 10.1073/pnas.87.17.6808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Trinchieri G. Interleukin-12 and its role in the generation of TH1 cells. Immunol Today. 1993 Jul;14(7):335–338. doi: 10.1016/0167-5699(93)90230-I. [DOI] [PubMed] [Google Scholar]
  35. Valiante N. M., Rengaraju M., Trinchieri G. Role of the production of natural killer cell stimulatory factor (NKSF/IL-12) in the ability of B cell lines to stimulate T and NK cell proliferation. Cell Immunol. 1992 Nov;145(1):187–198. doi: 10.1016/0008-8749(92)90322-g. [DOI] [PubMed] [Google Scholar]
  36. Wolf S. F., Temple P. A., Kobayashi M., Young D., Dicig M., Lowe L., Dzialo R., Fitz L., Ferenz C., Hewick R. M. Cloning of cDNA for natural killer cell stimulatory factor, a heterodimeric cytokine with multiple biologic effects on T and natural killer cells. J Immunol. 1991 May 1;146(9):3074–3081. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES