Abstract
The cell-surface expression of the class I alloantigen Qa-2 was analyzed on resting and activated spleen and thymus cells using cytotoxic elimination and immunofluorescence and flow cytometry. Spleen cells activated by mitogens or alloantigen were homogeneously positive for cell surface Qa-2, but activated splenic T cells expressed only about one-third as much Qa-2 per cell as did nonstimulated T cells. These data correlated with the ability to perform cytotoxic elimination with Qa-2-specific monoclonal antibodies (mAbs) in that cytotoxic T lymphocyte (CTL) activity was completely abrogated by pretreatment of spleen cells prior to in vitro culture but was only partially eliminated by treatment of CTL effectors. Qa-2-positive cells constituted only a small subpopulation of fresh normal thymocytes, but were enriched (>40% positive) among cortisone-resistant thymocytes (CRT). These Qa-2-positive CRT contained mature thymocytes as defined by Ly phenotype Ly-2−, Ly-1hi. When normal thymocytes were treated with Qa-2-specific mAb and complement prior to in vitro sensitization for generation of allogeneic CTL, CTL activity was completely abrogated despite the fact that the fraction of cells eliminated were undetectable as assessed by cell recovery. CTL effectors from alloantigen-stimulated thymocytes were also susceptible to cytotoxic elimination with Qa-2-specific mAb. These data suggest that the Qa-2 molecule may serve not only as a marker on resting and activated peripheral T cells, but also as a unique marker for functionally mature T cells in the thymus.
Similar content being viewed by others
References
Auchincloss, H., Jr., Ozato, K., and Sachs, D. H.: Two distinct murine differentiation antigens determined by genes linked to the Ly-6 locus. J. Immunol. 127: 1839–1843, 1981
Auchincloss, H., Jr., Ozato, K., and Sachs, D. H.: A monoclonal antibody detecting unusual Thy-1 determinants. J. Immunol. 128: 1584–1589, 1982
Ceredig, R., Glasebrook, A. L., and MacDonald, H. R.: Phenotypic and functional properties of murine thymocytes. I. Precursors of cytolytic T lymphocytes and interleukin 2-producing cells are all contained within a subpopulation of “mature” thymocytes as analyzed by monoclonal antibodies and flow microfluorometry. J. Exp. Med. 155: 358–379, 1982
Emerson, S. G., Reilly, P., and Cone, R. E.: Differential radiolabelling of lymphocyte membrane alloantigens and immunoglobulins: Variation of H2O2 concentration during lactoperoxidase catalyzed cell surface radio-iodination. J. Immunogenet. 6: 87–97, 1979
Flaherty, L.: The Tla region of the mouse: Identification of a new serologically defined locus, Qa-2. Immunogenetics 3: 533–539, 1976
Flaherty, L.: Tla-region antigens. In M. E. Dorf (ed.): The Role of the Major Histocompatibility Complex in Immunobiology, pp. 33–57, Garland STPM Press, New York, 1981
Flaherty, L., Zimmerman, D., and Sullivan, K. A.: Qa-2 and Qa-3 antigens on lymphocyte subpopulations. I. Mitogen responsiveness. J. Immunol. 121: 1640–1643, 1978
Fowlkes, B. J.: Characterization and Differentiation of Thymic lymphocytes in the Mouse. Ph. D. thesis, George Washington University Graduate School of Arts and Sciences, Washington, DC, 1984
Hämmerling, G. J., Hämmerling, U., and Flaherty, L.: Qat-4 and Qat-5, new murine T -cell antigens governed by the Tla region and identified by monoclonal antibodies. J. Exp. Med. 150: 108–116, 1979
Hogarth, P. M., Basten, A., Prichard-Briscoe, H., Henning, M. H., Sutton, V. R., and McKenzie, I. F. C.: The distribution of the Qa-2 alloantigen on functional T lymphocytes. J. Immunol. 135: 1632–1636, 1985
Hood, L., Steinmetz, M., and Malissen, B.: Genes of the major histocompatibility complex of the mouse. Annu. Rev. Immunol. 1: 529–557, 1983
Klein, J.: Biology of the Mouse Histocompatibility Complex, Springer-Verlag, New York, 1975
Kruisbeek, A. M., Glystra, J. J., and Krose, T. J. M.: Distinct effects of T cell growth factors and thymocyte epithelial factors on the generation of cytotoxic T lymphocytes by thymocyte subpopulations. J Immunol. 125: 995–1002, 1980
Laemmli, U. K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970
Landolfi, N. G., Soloski, M. J., and Cook, R. G.: Molecular weight diversity among murine class I antigens: Both the mature cell surface forms and the unglycosylated polypeptides vary significantly in molecular weight. Mol. Immunol. 22: 155–158, 1985
Mathieson, B. J. and Fowlkes, B. J.: Cell surface antigen expression on thymocytes: Development and phenotypic differentiation of interthymic subsets. Immunol. Rev. 82: 141–173, 1984
Mellor, A. L., Antoniou, J., and Robinson, P. J.: Structure and expression of genes encoding murine Qa-2 class I antigens. Proc. Natl. Acad. Sci. U.S.A. 82: 5920–5924, 1985
Michaelson, J., Flaherty, L., Hutchinson, B., and Yudkowitz, H.: Qa-2 does not display structural genetic polymorphism detectable on isoelectric-focusing gels. Immunogenetics 16: 363–366, 1982
Oudshoorn-Snoek, M., Mellor, A. L., Flavell, R. A., and Démant, P.: A new determinant, Qa-m208, detected on T lymphocytes and transfected L cells by a Kb-specific monoclonal antibody. Immunogenetics 19: 461–474, 1984
Ozato, K. and 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. 126: 317–321, 1981
Ozato, K., Mayer, N. M., and Sachs, D. H.: Monoclonal antibodies to mouse major histocompatibility complex antigens. IV. A series of hybridoma clones producing anti-H-2 antibodies and an examination of expression of H-2d antigens on the surface of these cells. Transplantation 34: 113–120, 1982
Rucker, J., Horowitz, M., Lerner, E. A., and Murphy, D. B.: Monoclonal antibody reveals H-2-linked quantitative and qualitative variation in the expression of a Qa-2 region determinant. Immunogenetics 17: 303–316, 1983
Sachs, D. H.: The major histocompatibility complex. In W. E. Paul (ed.): Fundamental Immunology, pp. 303–346, Raven Press, New York, 1984
Sandrin, M. S., Hogarth, P. M., and McKenzie, I. F. C.: Two “Qa” specificities: Qa-m7 and Qa-m8 defined by monoclonal antibodies. J. Immunol. 131: 546–547, 1983
Scollay, R., Jacobs, S., Jerabek, L., Butcher, E., and Weissman, I.: T cell maturation: Thymocytes and thymus migrant subpopulations defined with monoclonal antibodies to MHC region antigens. J. Immunol. 124: 2845–2853, 1980
Scollay, R., Bartlett, P., and Shortman, K.: T cell development in the adult murine thymus: Changes in expression of the surface antigens Ly2, Lyt4 and B2A2 during development from early precursor cells to migrants. Immunol. Rev. 82: 79–103, 1984
Segal, D. M., Sharrow, S. O., and Segal, D. M.: Fe(IgG) receptors on rat basophilic leukemia cells. J. Immunol. 126: 138–145, 1981
Sharrow, S. O., Mathieson, B. J., and Singer, A.: Cell surface appearance of unexpected host MHC determinants on thymocytes from radiation bone marrow chimeras. J. Immunol. 126: 1327–1335, 1981
Sharrow, S. O., Flaherty, L., and Sachs, D. H.: Serologic cross-reactivity between class I MHC molecules and an H-2-linked differentiation antigen as detected by monoclonal antibodies. J. Exp. Med. 159: 21–40, 1984
Sherman, D. H., Kranz, D. M., and Eisen, H. N.: Expression of structurally diverse Qa-2-encoded molecules on the surface of cloned cytotoxic T lymphocytes. J. Exp. Med. 160: 1421–1430, 1984
Soloski, M. J., Vernachio, J., Einhorn, G., and Lattimore, A.: Qa gene expression: Biosynthesis and secretion of Qa-2 molecules in activated T cells. Proc. Natl. Acad. Sci. U.S.A. 83: 2949–2953, 1986
Stroynowski, I., Forman, J., Goodenow, R. S., Schiffer, S. G., McMillan, M., Sharrow, S. O., Sachs, D. H., and Hood, L.: Expression and T cell recognition of hybrid antigens with amino-terminal domains encoded by Qa-2 region of major histocompatibility complex and carboxyl termini of transplantation antigens. J. Exp. Med. 161: 935–152, 1985
Sullivan, K. A. and Flaherty, L.: The Qa-2 antigen on lymphocyte subpopulations. Mixed lymphocyte cultures and cell-mediated lympholysis. J. Immunol. 123: 2920–2924, 1979
Titus, J. A., Haugland, R., Sharrow, S. O., and Segal, D. M.: Texas Red, a hydrophilic, red-emitting fluorophore for use with fluorescein in dual parameter flow microfluorometric and fluorescence microscope studies. J. Immunol. Methods 50: 193–204, 1982
Widacki, S., Flaherty, L., and Cook, R. G.: Biochemical characterization of the molecules reactive with Qa-6 antiserum and the monoclonal antibody 20-8-4: Evidence for structural similarity with the Qa-2 molecule. J. Immunol. 135: 3333–3339, 1985
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Rabinowitz, R., Sharrow, S.O., Chatterjee-Das, S. et al. Qa alloantigen expression on functional T lymphocytes from spleen and thymus. Immunogenetics 24, 391–401 (1986). https://doi.org/10.1007/BF00377958
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00377958