Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1984 Sep 1;160(3):799–813. doi: 10.1084/jem.160.3.799

Analysis of thymocyte MHC specificity with thymocyte hybridomas

PMCID: PMC2187409  PMID: 6206189

Abstract

Medullary, peanut agglutinin-negative (PNA-), thymocytes were activated in vitro with either exogenous interleukin 1 (IL-1) or accessory cells. T cell blasts from these cultures were subsequently fused to BW5147 to generate thymocyte hybridomas. Fusion frequencies similar to those obtained with peripheral T lymphocytes were observed. A high frequency of these hybrids are triggered to produce IL-2 in the presence of syngeneic accessory cells. Exogenous, nominal antigens do not appear to be required for this activation. Using accessory cells from a series of recombinant inbred mice, the specificity of this hybrid-accessory cell interaction could be mapped to either I-Ak or I-Ek or both. This was confirmed by blocking with alpha Ia monoclonal antibodies (mAbs). A high frequency of these self-reactive cells are also alloreactive. Interestingly, several clones were identified that appear to recognize public Ia determinants broadly shared by different alleles and genetic subregions. Such specificities appear to contrast with those of peripheral T lymphocytes whose specificity is dominated by the genetically polymorphic portion of the Ia molecule. These results document the clonal specificity occurring in the cultures of in vitro activated thymocytes and allow an analysis of at least a portion of the intrathymic repertoire for major histocompatibility complex (MHC) determinants. The implications of these findings are discussed.

Full Text

The Full Text of this article is available as a PDF (922.1 KB).

Selected References

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

  1. Bevan M. J., Fink P. J. The influence of thymus H-2 antigens on the specificity of maturing killer and helper cells. Immunol Rev. 1978;42:3–19. doi: 10.1111/j.1600-065x.1978.tb00256.x. [DOI] [PubMed] [Google Scholar]
  2. Bhattacharya A., Dorf M. E., Springer T. A. A shared alloantigenic determinant on Ia antigens encoded by the I-A and I-E subregions: evidence for I region gene duplication. J Immunol. 1981 Dec;127(6):2488–2495. [PubMed] [Google Scholar]
  3. Bruce J., Symington F. W., McKearn T. J., Sprent J. A monoclonal antibody discriminating between subsets of T and B cells. J Immunol. 1981 Dec;127(6):2496–2501. [PubMed] [Google Scholar]
  4. Fathman C. G., Hengartner H. Crossreactive mixed lymphocyte reaction determinants recognized by cloned alloreactive T cells. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5863–5866. doi: 10.1073/pnas.76.11.5863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Finberg R., Burakoff S. J., Cantor H., Benacerraf B. Biological significance of alloreactivity: T cells stimulated by Sendai virus-coated syngeneic cells specifically lyse allogeneic target cells. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5145–5149. doi: 10.1073/pnas.75.10.5145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Glimcher L. H., Schwartz R. H., Longo D. L., Singer A. The specificity of the syngeneic mixed leukocyte response, a primary anti-I region T cell proliferative response, is determined intrathymically. J Immunol. 1982 Sep;129(3):987–991. [PubMed] [Google Scholar]
  7. Glimcher L. H., Shevach E. M. Production of autoreactive I region-restricted T cell hybridomas. J Exp Med. 1982 Aug 1;156(2):640–645. doi: 10.1084/jem.156.2.640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Haskins K., Kubo R., White J., Pigeon M., Kappler J., Marrack P. The major histocompatibility complex-restricted antigen receptor on T cells. I. Isolation with a monoclonal antibody. J Exp Med. 1983 Apr 1;157(4):1149–1169. doi: 10.1084/jem.157.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Heber-Katz E., Schwartz R. H., Matis L. A., Hannum C., Fairwell T., Appella E., Hansburg D. Contribution of antigen-presenting cell major histocompatibility complex gene products to the specificity of antigen-induced T cell activation. J Exp Med. 1982 Apr 1;155(4):1086–1099. doi: 10.1084/jem.155.4.1086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Janeway C. A., Jr, Conrad P. J., Lerner E. A., Babich J., Wettstein P., Murphy D. B. Monoclonal antibodies specific for Ia glycoproteins raised by immunization with activated T cells: possible role of T cellbound Ia antigens as targets of immunoregulatory T cells. J Immunol. 1984 Feb;132(2):662–667. [PubMed] [Google Scholar]
  11. Kappler J. W., Marrack P. The role of H-2 linked genes in helper T-cell function. IV. Importance of T-cell genotype and host environment in I-region and Ir gene expression. J Exp Med. 1978 Dec 1;148(6):1510–1522. doi: 10.1084/jem.148.6.1510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kappler J. W., Skidmore B., White J., Marrack P. Antigen-inducible, H-2-restricted, interleukin-2-producing T cell hybridomas. Lack of independent antigen and H-2 recognition. J Exp Med. 1981 May 1;153(5):1198–1214. doi: 10.1084/jem.153.5.1198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kaufman J. F., Auffray C., Korman A. J., Shackelford D. A., Strominger J. The class II molecules of the human and murine major histocompatibility complex. Cell. 1984 Jan;36(1):1–13. doi: 10.1016/0092-8674(84)90068-0. [DOI] [PubMed] [Google Scholar]
  14. Kruisbeek A. M., Astaldi G. C. Distinct effects of thymic epithelial culture supernatants on T cell properties of mouse thymocytes separated by the use of peanut agglutinin. J Immunol. 1979 Sep;123(3):984–991. [PubMed] [Google Scholar]
  15. Marshak-Rothstein A., Fink P., Gridley T., Raulet D. H., Bevan M. J., Gefter M. L. Properties and applications of monoclonal antibodies directed against determinants of the Thy-1 locus. J Immunol. 1979 Jun;122(6):2491–2497. [PubMed] [Google Scholar]
  16. 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]
  17. Ozato K., Mayer N., Sachs D. H. Hybridoma cell lines secreting monoclonal antibodies to mouse H-2 and Ia antigens. J Immunol. 1980 Feb;124(2):533–540. [PubMed] [Google Scholar]
  18. Rock K. L., Benacerraf B. MHC-restricted T cell activation: analysis with T cell hybridomas. Immunol Rev. 1983;76:29–57. doi: 10.1111/j.1600-065x.1983.tb01096.x. [DOI] [PubMed] [Google Scholar]
  19. Rock K. L., Benacerraf B. Selective modification of a private I-A allo-stimulating determinant(s) upon association of antigen with an antigen-presenting cell. J Exp Med. 1984 Apr 1;159(4):1238–1252. doi: 10.1084/jem.159.4.1238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rock K. L., Benacerraf B. The role of Ia molecules in the activation of T lymphocytes. IV. The basis of the thymocyte IL 1 response and its possible role in the generation of the T cell repertoire. J Immunol. 1984 Apr;132(4):1654–1662. [PubMed] [Google Scholar]
  21. Rock K. L., Benacerraf B. Thymic T cells are driven to expand upon interaction with self-class II major histocompatibility complex gene products on accessory cells. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1221–1224. doi: 10.1073/pnas.81.4.1221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rosenthal A. S., Shevach E. M. Function of macrophages in antigen recognition by guinea pig T lymphocytes. I. Requirement for histocompatible macrophages and lymphocytes. J Exp Med. 1973 Nov 1;138(5):1194–1212. doi: 10.1084/jem.138.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shevach E. M., Rosenthal A. S. Function of macrophages in antigen recognition by guinea pig T lymphocytes. II. Role of the macrophage in the regulation of genetic control of the immune response. J Exp Med. 1973 Nov 1;138(5):1213–1229. doi: 10.1084/jem.138.5.1213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sredni B., Schwartz R. H. Alloreactivity of an antigen-specific T-cell clone. Nature. 1980 Oct 30;287(5785):855–857. doi: 10.1038/287855a0. [DOI] [PubMed] [Google Scholar]
  25. Steinman R. M., Cohn Z. A. Identification of a novel cell type in peripheral lymphoid organs of mice. II. Functional properties in vitro. J Exp Med. 1974 Feb 1;139(2):380–397. doi: 10.1084/jem.139.2.380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Von Boehmer H., Adams P. B. Syngeneic mixed lymphocyte reaction between thymocytes and peripheral lymphoid cells in mice: strain specificity and nature of the target cell. J Immunol. 1973 Feb;110(2):376–383. [PubMed] [Google Scholar]

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

RESOURCES