Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1985 Sep 1;162(3):943–961. doi: 10.1084/jem.162.3.943

Characterization of antigen-specific, Ia-restricted, L3T4+ cytolytic T lymphocytes and assessment of thymic influence on their self specificity

PMCID: PMC2187814  PMID: 2411844

Abstract

The goals of the present study were: (a) to generate antigen-specific L3T4+ cytolytic T lymphocytes (CTL), (b) to determine their major histocompatibility complex (MHC) restriction specificity, and (c) to assess the influence of thymic MHC determinants on their self specificity. We found that L3T4+ CTL specific for either trinitrophenyl (TNP)-modified self determinants or minor histocompatibility antigens could be generated from Lyt-2- responder T cells provided that the response cultures were supplemented with supernatants rich in helper factors. Such antigen-specific L3T4+ CTL were Ia-restricted by the criteria that they lysed only Ia+ target cells and that their lysis of Ia+ target cells was specifically inhibited by anti-Ia monoclonal antibodies. The relative frequency of L3T4+ pCTL was found to be only 5- 10% of the total anti-TNP pCTL present in the spleens of normal mice. Finally, we utilized radiation bone marrow chimeras to assess the influence of the thymic haplotype on the self-Ia specificity of L3T4+ CTL. Both bulk culture and limiting dilution experiments revealed that the self-Ia specificity of L3T4+ anti-TNP CTL from F1----parent and A--- -B allogeneic chimeras was not markedly skewed toward the haplotype of the chimeric thymus. These results contrast with those obtained previously for L3T4+ anti-TNP Th cells and demonstrate that in the radiation bone marrow chimera model of T cell differentiation, the self specificity of Th cells but not pCTL is markedly influenced by the haplotype of the chimeric thymus.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

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

  1. Bevan M. J. The major histocompatibility complex determines susceptibility to cytotoxic T cells directed against minor histocompatibility antigens. J Exp Med. 1975 Dec 1;142(6):1349–1364. doi: 10.1084/jem.142.6.1349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradley S. M., Kruisbeek A. M., Singer A. Cytotoxic T lymphocyte responses in allogeneic radiation bone marrow chimeras. The chimeric host strictly dictates the self-repertoire of Ia-restricted T cells but not H-2K/D-restricted T cells. J Exp Med. 1982 Dec 1;156(6):1650–1664. doi: 10.1084/jem.156.6.1650. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dialynas D. P., Quan Z. S., Wall K. A., Pierres A., Quintáns J., Loken M. R., Pierres M., Fitch F. W. Characterization of the murine T cell surface molecule, designated L3T4, identified by monoclonal antibody GK1.5: similarity of L3T4 to the human Leu-3/T4 molecule. J Immunol. 1983 Nov;131(5):2445–2451. [PubMed] [Google Scholar]
  4. Fink P. J., Bevan M. J. H-2 antigens of the thymus determine lymphocyte specificity. J Exp Med. 1978 Sep 1;148(3):766–775. doi: 10.1084/jem.148.3.766. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Glimcher L. H., Kim K. J., Green I., Paul W. E. Ia antigen-bearing B cell tumor lines can present protein antigen and alloantigen in a major histocompatibility complex-restricted fashion to antigen-reactive T cells. J Exp Med. 1982 Feb 1;155(2):445–459. doi: 10.1084/jem.155.2.445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Guillemot F. P., Oliver P. D., Peault B. M., Le Douarin N. M. Cells expressing Ia antigens in the avian thymus. J Exp Med. 1984 Dec 1;160(6):1803–1819. doi: 10.1084/jem.160.6.1803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hayday A. C., Saito H., Gillies S. D., Kranz D. M., Tanigawa G., Eisen H. N., Tonegawa S. Structure, organization, and somatic rearrangement of T cell gamma genes. Cell. 1985 Feb;40(2):259–269. doi: 10.1016/0092-8674(85)90140-0. [DOI] [PubMed] [Google Scholar]
  8. Hodes R. J., Singer A. Cellular and genetic control of antibody responses in vitro. I. Cellular requirements for the generation of genetically controlled primary IgM responses to soluble antigens. Eur J Immunol. 1977 Dec;7(12):892–897. doi: 10.1002/eji.1830071214. [DOI] [PubMed] [Google Scholar]
  9. Jacobson S., Nepom G. T., Richert J. R., Biddison W. E., McFarland H. F. Identification of a specific HLA DR2 Ia molecule as a restriction element for measles virus-specific HLA class II-restricted cytotoxic T cell clones. J Exp Med. 1985 Jan 1;161(1):263–268. doi: 10.1084/jem.161.1.263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jacobson S., Richert J. R., Biddison W. E., Satinsky A., Hartzman R. J., McFarland H. F. Measles virus-specific T4+ human cytotoxic T cell clones are restricted by class II HLA antigens. J Immunol. 1984 Aug;133(2):754–757. [PubMed] [Google Scholar]
  11. Kast W. M., de Waal L. P., Melief C. J. Thymus dictates major histocompatibility complex (MHC) specificity and immune response gene phenotype of class II MHC-restricted T cells but not of class I MHC-restricted T cells. J Exp Med. 1984 Dec 1;160(6):1752–1766. doi: 10.1084/jem.160.6.1752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kruisbeek A. M., Andrysiak P. M., Singer A. Self recognition of accessory cell Ia determinants is required for the in vitro generation of hapten-specific cytotoxic T lymphocyte responses. J Immunol. 1983 Oct;131(4):1650–1655. [PubMed] [Google Scholar]
  13. Kruisbeek A. M., Fultz M. J., Sharrow S. O., Singer A., Mond J. J. Early development of the T cell repertoire. In vivo treatment of neonatal mice with anti-Ia antibodies interferes with differentiation of I-restricted T cells but not K/D-restricted T cells. J Exp Med. 1983 Jun 1;157(6):1932–1946. doi: 10.1084/jem.157.6.1932. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kruisbeek A. M., Sharrow S. O., Mathieson B. J., Singer A. The H-2 phenotype of the thymus dictates the self-specificity expressed by thymic but not splenic cytotoxic T lymphocyte precursors in thymus-engrafted nude mice. J Immunol. 1981 Nov;127(5):2168–2176. [PubMed] [Google Scholar]
  15. Kruisbeek A. M., Sharrow S. O., Singer A. Differences in the MHC-restricted self-recognition repertoire of intra-thymic and extra-thymic cytotoxic T lymphocyte precursors. J Immunol. 1983 Mar;130(3):1027–1032. [PubMed] [Google Scholar]
  16. Kruisbeek A. M., Zijlstra J. J., Kröse T. J. Distinct effects of T cell growth factors and thymic epithelial factors on the generation of cytotoxic T lymphocytes by thymocyte subpopulations. J Immunol. 1980 Sep;125(3):995–1002. [PubMed] [Google Scholar]
  17. Ly I. A., Mishell R. I. Separation of mouse spleen cells by passage through columns of sephadex G-10. J Immunol Methods. 1974 Aug;5(3):239–247. doi: 10.1016/0022-1759(74)90108-2. [DOI] [PubMed] [Google Scholar]
  18. Mizuochi T., Golding H., Rosenberg A. S., Glimcher L. H., Malek T. R., Singer A. Both L3T4+ and Lyt-2+ helper T cells initiate cytotoxic T lymphocyte responses against allogenic major histocompatibility antigens but not against trinitrophenyl-modified self. J Exp Med. 1985 Aug 1;162(2):427–443. doi: 10.1084/jem.162.2.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Oi V. T., Jones P. P., Goding J. W., Herzenberg L. A., Herzenberg L. A. Properties of monoclonal antibodies to mouse Ig allotypes, H-2, and Ia antigens. Curr Top Microbiol Immunol. 1978;81:115–120. doi: 10.1007/978-3-642-67448-8_18. [DOI] [PubMed] [Google Scholar]
  20. Ozato K., Sachs D. H. Monoclonal antibodies to mouse MHC antigens. III. Hybridoma antibodies reacting to antigens of the H-2b haplotype reveal genetic control of isotype expression. J Immunol. 1981 Jan;126(1):317–321. [PubMed] [Google Scholar]
  21. Pierres A., Naquet P., Van Agthoven A., Bekkhoucha F., Denizot F., Mishal Z., Schmitt-Verhulst A. M., Pierres M. A rat anti-mouse T4 monoclonal antibody (H129.19) inhibits the proliferation of Ia-reactive T cell clones and delineates two phenotypically distinct (T4+, Lyt-2,3-, and T4-, Lyt-2,3+) subsets among anti-Ia cytolytic T cell clones. J Immunol. 1984 Jun;132(6):2775–2782. [PubMed] [Google Scholar]
  22. Pober J. S., Gimbrone M. A., Jr, Cotran R. S., Reiss C. S., Burakoff S. J., Fiers W., Ault K. A. Ia expression by vascular endothelium is inducible by activated T cells and by human gamma interferon. J Exp Med. 1983 Apr 1;157(4):1339–1353. doi: 10.1084/jem.157.4.1339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Raulet D. H., Bevan M. J. Helper T cells for cytotoxic T lymphocytes need not be I region restricted. J Exp Med. 1982 Jun 1;155(6):1766–1784. doi: 10.1084/jem.155.6.1766. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shearer G. M. Cell-mediated cytotoxicity to trinitrophenyl-modified syngeneic lymphocytes. Eur J Immunol. 1974 Aug;4(8):527–533. doi: 10.1002/eji.1830040802. [DOI] [PubMed] [Google Scholar]
  25. Shiku H., Kisielow P., Bean M. A., Takahashi T., Boyse E. A., Oettgen H. F., Old L. J. Expression of T-cell differentiation antigens on effector cells in cell-mediated cytotoxicity in vitro. Evidence for functional heterogeneity related to the surface phenotype of T cells. J Exp Med. 1975 Jan 1;141(1):227–241. doi: 10.1084/jem.141.1.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Singer A., Hathcock K. S., Hodes R. J. Self recognition in allogeneic radiation bone marrow chimeras. A radiation-resistant host element dictates the self specificity and immune response gene phenotype of T-helper cells. J Exp Med. 1981 May 1;153(5):1286–1301. doi: 10.1084/jem.153.5.1286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Singer A., Hathcock K. S., Hodes R. J. Self recognition in allogeneic thymic chimeras. Self recognition by T helper cells from thymus-engrafted nude mice is restricted to the thymic H-2 haplotype. J Exp Med. 1982 Jan 1;155(1):339–344. doi: 10.1084/jem.155.1.339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sprent J. Restricted helper function of F1 leads to parent bone marrow chimeras controlled by K-end of H-2 complex. J Exp Med. 1978 Jun 1;147(6):1838–1842. doi: 10.1084/jem.147.6.1838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Taswell C. Limiting dilution assays for the determination of immunocompetent cell frequencies. I. Data analysis. J Immunol. 1981 Apr;126(4):1614–1619. [PubMed] [Google Scholar]
  30. Tite J. P., Janeway C. A., Jr Cloned helper T cells can kill B lymphoma cells in the presence of specific antigen: Ia restriction and cognate vs. noncognate interactions in cytolysis. Eur J Immunol. 1984 Oct;14(10):878–886. doi: 10.1002/eji.1830141004. [DOI] [PubMed] [Google Scholar]
  31. Wagner H., Hardt C., Stockinger H., Pfizenmaier K., Bartlett R., Röllinghoff M. Impact of thymus on the generation of immunocompetence and diversity of antigen-specific MHC-restricted cytotoxic T-lymphocyte precursors. Immunol Rev. 1981;58:95–129. doi: 10.1111/j.1600-065x.1981.tb00351.x. [DOI] [PubMed] [Google Scholar]
  32. Zinkernagel R. M., Callahan G. N., Althage A., Cooper S., Klein P. A., Klein J. On the thymus in the differentiation of "H-2 self-recognition" by T cells: evidence for dual recognition? J Exp Med. 1978 Mar 1;147(3):882–896. doi: 10.1084/jem.147.3.882. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES