Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1990 Apr 1;171(4):1057–1071. doi: 10.1084/jem.171.4.1057

Cytotoxic T lymphocytes reactive against a syngeneic murine tumor and their specific suppressor T cells are both elicited by in vitro allosensitization

PMCID: PMC2187835  PMID: 2139097

Abstract

Sensitization of C57BL/6 (B6, H-2b) splenocytes against normal BALB/c (H-2d) leukocytes (B6 a/BALB) in bulk MLC induced CTL reactive against the syngeneic (H-2b) nonimmunogenic lymphoma PIR-2, in addition to the CTL directed against the corresponding (H-2d) allotargets. However, MLC- derived lymphocytes did not directly exhibit anti-PIR-2 cytotoxicity in spite of the high anti-PIR-2 CTL frequency (up to 1/20) among them, as demonstrated by the limiting dilution culture (LDC) technique. The present study was undertaken to resolve this contradiction. We found that anti-PIR-2 cytotoxicity could be detected only when B6 a/BALB MLC- derived responding cells were plated in LDC at low numbers (less than 200) of cells/well. In contrast, increasing the number of the plated cells to 500-5,000 resulted in a gradual decrease in the percentage of wells cytotoxically reactive against PIR-2, whereas the percentage of wells exhibiting cytotoxicity against the allotargets remained unchanged (100%). This decrease of anti-PIR-2 cytotoxicity in LDC and the lack of anti-PIR-2 reactivity among MLC-derived lymphocytes were shown by mixing experiments to result from the activity of radioresistant Thy-1+, Lyt-2+, L3T4- suppressor cells, blocking the anti-PIR-2 cytotoxicity at the effector phase. The suppression was specific as indicated by the following observations: (a) freshly obtained B6 splenocytes, cultured unsensitized B6 splenocytes, mitogen- induced B6 lymphoblasts, B6 LAK cells, or B6 a/B6 MLC-derived lymphocytes were not suppressive; (b) anti-PIR-2 cytotoxicity elicited in B6 a/BALB LDC was suppressed only by lymphocytes derived from B6 a/BALB MLC and not from B6 a/C3H (H-2k) MLC; and (c) B6 a/BALB MLC- induced suppressor cells could be adsorbed on monolayers of BALB/c but not of C3H lymphoblasts. Since both syngeneic tumor and allogeneic target cells were lysed by the same clonal cell population but only the antisyngeneic activity was suppressed, we suggest that a single CTL can exhibit two cytotoxic activities that are differentially affected by the described suppressor cells. This mode of suppression may play a role in controlling autoimmune reactivity.

Full Text

The Full Text of this article is available as a PDF (981.1 KB).

Selected References

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

  1. Altman A., Katz D. H. Existence of T cells manifesting self-reactivity indistinguishable from alloreactivity. J Immunol. 1980 Oct;125(4):1536–1543. [PubMed] [Google Scholar]
  2. Ben-Nun A., Lando Z., Dorf M. E., Burakoff S. J. Analysis of cross-reactive antigen-specific T cell clones. Specific recognition of two major histocompatibility complex (MHC) and two non-MHC antigens by a single clone. J Exp Med. 1983 Jun 1;157(6):2147–2153. doi: 10.1084/jem.157.6.2147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brooks C. G., Urdal D. L., Henney C. S. Lymphokine-driven "differentiation" of cytotoxic T-cell clones into cells with NK-like specificity: correlations with display of membrane macromolecules. Immunol Rev. 1983;72:43–72. doi: 10.1111/j.1600-065x.1983.tb01072.x. [DOI] [PubMed] [Google Scholar]
  4. Cerottini J. C., Brunner K. T. Cell-mediated cytotoxicity, allograft rejection, and tumor immunity. Adv Immunol. 1974;18:67–132. doi: 10.1016/s0065-2776(08)60308-9. [DOI] [PubMed] [Google Scholar]
  5. Cohen I. R. Regulation of autoimmune disease physiological and therapeutic. Immunol Rev. 1986 Dec;94:5–21. doi: 10.1111/j.1600-065x.1986.tb01161.x. [DOI] [PubMed] [Google Scholar]
  6. Corley R. B., Kindred B., Lefkovits I. Positive and negative allogeneic effects mediated by MLR-primed lymphocytes: quantitation by limiting dilution analysis. J Immunol. 1978 Sep;121(3):1082–1089. [PubMed] [Google Scholar]
  7. Duke R. C. Self recognition by T cells. I. Bystander killing of target cells bearing syngeneic MHC antigens. J Exp Med. 1989 Jul 1;170(1):59–71. doi: 10.1084/jem.170.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ellerman K. E., Powers J. M., Brostoff S. W. A suppressor T-lymphocyte cell line for autoimmune encephalomyelitis. Nature. 1988 Jan 21;331(6153):265–267. doi: 10.1038/331265a0. [DOI] [PubMed] [Google Scholar]
  9. Eura M., Maehara T., Ikawa T., Ishikawa T. Suppressor cells in the effector phase of autologous cytotoxic reactions in cancer patients. Cancer Immunol Immunother. 1988;27(2):147–153. doi: 10.1007/BF00200020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fink P. J., Rammensee H. G., Benedetto J. D., Staerz U. D., Lefrancois L., Bevan M. J. Studies on the mechanism of suppression of primary cytotoxic responses by cloned cytotoxic T lymphocytes. J Immunol. 1984 Oct;133(4):1769–1774. [PubMed] [Google Scholar]
  11. Goronzy J., Schaefer U., Eichmann K., Simon M. M. Quantitative studies on T cell diversity. II. Determination of the frequencies and Lyt phenotypes of two types of precursor cells for alloreactive cytotoxic T cells in polyclonally and specifically activated splenic T cells. J Exp Med. 1981 Apr 1;153(4):857–870. doi: 10.1084/jem.153.4.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Joly P., Guillon J. M., Mayaud C., Plata F., Theodorou I., Denis M., Debre P., Autran B. Cell-mediated suppression of HIV-specific cytotoxic T lymphocytes. J Immunol. 1989 Oct 1;143(7):2193–2201. [PubMed] [Google Scholar]
  13. Kotani H., Mitsuya H., Benson E., James S. P., Strober W. Activation and function of an autoreactive T cell clone with dual immunoregulatory activity. J Immunol. 1988 Jun 15;140(12):4167–4172. [PubMed] [Google Scholar]
  14. Leshem B., Epstein E., Kedar E. Allosensitization in IL-2-containing limiting dilution cultures generates cytotoxic T lymphocytes (CTL) rather than LAK cells reactive with a syngeneic nonimmunogenic murine tumor. Immunol Lett. 1989 Jan 15;20(1):53–58. doi: 10.1016/0165-2478(89)90068-0. [DOI] [PubMed] [Google Scholar]
  15. Leshem B., Gotsman B., Kedar E. In vitro elicitation of cytotoxic response against a nonimmunogenic murine tumor by allosensitization. Cancer Immunol Immunother. 1984;17(2):117–123. doi: 10.1007/BF00200047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. MacDonald H. R., Cerottini J. C., Ryser J. E., Maryanski J. L., Taswell C., Widmer M. B., Brunner K. T. Quantitation and cloning of cytolytic T lymphocytes and their precursors. Immunol Rev. 1980;51:93–123. doi: 10.1111/j.1600-065x.1980.tb00318.x. [DOI] [PubMed] [Google Scholar]
  17. Martin A., Eiermann T. H., Tricas L., Goldmann S. F. MHC class II restricted self-recognition after allostimulation: clonal analysis. Tissue Antigens. 1987 Jul;30(1):1–10. doi: 10.1111/j.1399-0039.1987.tb01588.x. [DOI] [PubMed] [Google Scholar]
  18. Mazumder A., Grimm E. A., Rosenberg S. A. Lysis of fresh human solid tumor cells by autologous lymphocytes activated in vitro by allosensitization. Cancer Immunol Immunother. 1983;15(1):1–10. doi: 10.1007/BF00199454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Miller R. G. An immunological suppressor cell inactivating cytotoxic T-lymphocyte precursor cells recognizing it. Nature. 1980 Oct 9;287(5782):544–546. doi: 10.1038/287544a0. [DOI] [PubMed] [Google Scholar]
  20. Nagarkatti P. S., Nagarkatti M., Mann L. W., Jones L. A., Kaplan A. M. Characterization of an endogenous Lyt2+ T-suppressor-cell population regulating autoreactive T cells in vitro and in vivo. Cell Immunol. 1988 Mar;112(1):64–77. doi: 10.1016/0008-8749(88)90276-6. [DOI] [PubMed] [Google Scholar]
  21. Ogasawara M., Iwabuchi K., Good R. A., Onoe K. Bone marrow cells from allogeneic bone marrow chimeras inhibit the generation of cytotoxic lymphocyte responses against both donor and recipient cells. J Immunol. 1988 Nov 15;141(10):3306–3312. [PubMed] [Google Scholar]
  22. Parmiani G., Sensi M. L., Carbone G., Colombo M., Pierotti M., Ballinari D., Hilgers J., Hilkens J. Cross-reactions between tumor cells and allogeneic normal tissues. Inhibition of a syngeneic lymphoma outgrowth in H-2 and non-H-2 alloimmune BALB/c mice. Int J Cancer. 1982 Mar 15;29(3):323–332. doi: 10.1002/ijc.2910290316. [DOI] [PubMed] [Google Scholar]
  23. Reimann J., Miller R. G. Generation of autoreactive cytotoxic T lymphocytes under limiting dilution conditions. J Immunol. 1983 Nov;131(5):2128–2134. [PubMed] [Google Scholar]
  24. 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]
  25. Rosenkrantz K., Dupont B., Flomenberg N. Generation and regulation of autocytotoxicity in mixed lymphocyte cultures: evidence for active suppression of autocytotoxic cells. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4508–4512. doi: 10.1073/pnas.82.13.4508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sakaguchi S., Fukuma K., Kuribayashi K., Masuda T. Organ-specific autoimmune diseases induced in mice by elimination of T cell subset. I. Evidence for the active participation of T cells in natural self-tolerance; deficit of a T cell subset as a possible cause of autoimmune disease. J Exp Med. 1985 Jan 1;161(1):72–87. doi: 10.1084/jem.161.1.72. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sensi M. L., Parenza M., Parmiani G. Alloreactivity and tumor antigens: generation of syngeneic antilymphoma killer lymphocytes by alloimmunization of mice with normal cells. J Natl Cancer Inst. 1983 Feb;70(2):291–297. [PubMed] [Google Scholar]
  28. Sensi M., Orosz C. G., Bach F. H. Alloantigen-induced cytotoxicity against syngeneic tumor cells: analysis at the clonal level. J Immunol. 1984 Jun;132(6):3218–3225. [PubMed] [Google Scholar]
  29. Shau H., Gray J. D., Golub S. H. Studies on cytotoxicity generated in human mixed lymphocyte culture. IV. Interleukin 2 alone or from mixed lymphocyte culture yields natural killer-like cytotoxic cells distinct from allospecific cytotoxic T lymphocytes. Cancer Immunol Immunother. 1988;27(3):255–260. doi: 10.1007/BF00205448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sheil J. M., Bevan M. J., Lefrancois L. Characterization of dual-reactive H-2Kb-restricted anti-vesicular stomatitus virus and alloreactive cytotoxic T cells. J Immunol. 1987 Jun 1;138(11):3654–3660. [PubMed] [Google Scholar]
  31. Theofilopoulos A. N., Dixon F. J. Etiopathogenesis of murine SLE. Immunol Rev. 1981;55:179–216. doi: 10.1111/j.1600-065x.1981.tb00343.x. [DOI] [PubMed] [Google Scholar]
  32. Vánky F., Gorsky T., Gorsky Y., Masucci M. G., Klein E. Lysis of tumor biopsy cells by autologous T lymphocytes activated in mixed cultures and propagated with T cell growth factor. J Exp Med. 1982 Jan 1;155(1):83–95. doi: 10.1084/jem.155.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Zarling J. M., Robins H. I., Raich P. C., Bach F. H., Bach M. L. Generation of cytotoxic T lymphocytes to autologous human leukaemia cells by sensitisation to pooled allogeneic normal cells. Nature. 1978 Jul 20;274(5668):269–271. doi: 10.1038/274269a0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES