Abstract
We report that mouse L cells expressing Ak class II molecules on their surface after DNA-mediated gene transfer are capable of presenting the synthetic copolymer (Glu60 Ala30 Tyr10) to Ak-restricted long-term T-cell clones. Antigen-induced T-cell stimulation could be inhibited by monoclonal antibodies (mAb) directed at spatially distinct determinants of the α and/or β subunits of the Ak molecule, and by the rat L3T4-specific mAb H129.19. In contrast, several rat mAb reactive with the mouse LFA-1 molecule failed to inhibit T-cell activation when L cells were used as antigen-presenting cells (APC), although these mAb strongly inhibited the same T-cell responses in the presence of leukocytic APC. Similarly, the cytolytic activity of the Ak-specific T-cell clone A15.1.17 was blocked by L3T4-specific and by LFA-1-specific mAb when tested on Ak-positive B-cell hybridomas, but only by L3T4-specific mAb and not by LFA-1-specific mAb when Ak-positive L-cell transformants were used as targets. These data support the notion that the LFA-1 molecule is not necessary for T-cell activation, and suggest that its functional role as an accessory molecule depends on the leukocytic nature of the APC tested.
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Beck, B. N., Glimcher, L. H., Nilson, A. E., Pierres, M., and McKean, D. J.: The structure-function relationship of I-Ak molecules: Correlation of serological and functional phenotypes of four I-Ak mutant cell lines. J. Immunol. 133: 3176–3182, 1984
Bekkhoucha, F., Naquet, P., Pierres, A., Marchetto, S., and Pierres, M.: Efficiency of antigen presentation to T cell clones by (B cell × B cell lymphoma) hybridomas correlates quantitatively with cell surface la antigen expression. Eur. J. Immunol. 14: 807–814, 1984
Ben-Nun, A., Glimcher, L. H., Weis, J., and Seidman J. F.: Functional expression of a cloned I-A kβ gene in B-lymphoma cells. Science 223: 825–827, 1984
Collins, T., Dreusky, A. M., Clayberger, C., Fiers, W., Gimbrone, M. A., Burakoff, S. J., Jr., and Pober, J. S.: Human cytolytic T lymphocyte interactions with vascular endothelium and fibroblasts: Role of effector and target cell molecules. J. Immunol. 133: 1878–1884, 1984
Davignon, D., Martz, E., Reynolds, T., Kurzinger, K., and Springer, T. A.: Lymphocyte function associated antigen-1 (LFA-1): A surface antigen distinct from Lyt-23 that participates in T lymphocyte mediated killing. Proc. Natl. Acad. Sci. U.S.A. 78: 4535–4539, 1981
Dialynas, D. P., Cohen, M. R., Sarmiento, M., and Fitch, F. W.: Identification of lysis-relevant molecules on the surfaces of CTL: Primary screening of monoclonal antibodies for the capacity to block cytolysis by cloned CTL lines. In W. R. Clark and P. Goldstein (eds.): Mechanisms of Cell-Mediated Cytotoxicity, pp. 547–556, Plenum, New York, 1982
Dialynas, D. P., Wilde, D. B., Marrack, P., Pierres, A., Wall, K. A., Havran, W., Otten, G., Loken, M. R., Pierres, M., Kappler, J., and Fitch, F. W.: Characterization of the murine antigenic determinant, designated L3T4a, recognized by monoclonal antibody GK1.5: Expression of L3T4a by functional T cell clones appears to correlate primarily with class II MHC antigen-reactivity. Immunol. Rev. 74: 29–56, 1983
Farr, A. E., Dorf, M. E., and Unanue, E. R.: Secretion of mediators following T-lymphocyte-macrophage interaction is regulated by the major histocompatibility complex. Proc. Natl. Acad. Sci. U.S.A. 74: 3542–3546, 1977
Germain, R. N., Norcross, M. A., and Margulies, D. H.: Functional expression of a transfected murine class II MHC gene. Nature 306: 190–194, 1983
Gillis, S. and Smith, K. A.: Long term culture of tumour-specific cytotoxic T cells. Nature 268: 154–156, 1977
Glimcher, L. H., Hamano, T., Asofsky, R., Sachs, D. H., Pierres, M., Samelson, L. E., Sharrow, S. O., and Paul, W. E.: Ia mutant functional antigen-presenting cell lines. J. Immunol. 130: 2287–2294, 1983
Golde, W.T., Kappler, J.W., Greenstein, J., Malissen, B., Hood, L., and Marrack, P.: The major histocompatibility complex-restricted antigen receptor on T cells. VIII. The role of the LFAJ molecule. J. Exp. Med. 161: 635–640, 1985
Golstein, P., Goridis, C., Schmitt-Verhulst, A. M., Hayot, B., Pierres, A., Van Agthoven, A., Kaufmann, Y., Esshar, Z., and Pierres, M.: Lymphoid cell surface interaction structures detected using cytolysis inhibiting monoclonal antibodies. Immunol. Rev. 68: 5–42, 1982
Greenstein, J. L., Marrack, P., Kappler, J., and Burakoff, S.: The role of L3T4 in recognition of la by a cytotoxic, H-2Dd specific T cell hybridoma. J. Exp. Med. 159: 1213–1224, 1984
Gromkowski, S. H., Heagy, W., and Martz, E.: Blocking of CTL-mediated killing by monoclonal antibodies to LFA-1 and Lyt-2,3. II. Evidence that trypsin pretreatment of target cells removes a non-H-2 molecule important in killing. J. Immunol. 134: 70–77, 1985
Hamano, T., Kim, K. J., Leisserson, W. M., and Asofsky, R.: Establishment of a B cell hybridoma with B cell surface antigens. J. Immunol. 129: 1403–1406, 1982
Malissen, B., Steinmetz, M., McMillan, M., Pierres, M., and Hood, L.: Expression of I-Ak class II genes in mouse L cells after DNA mediated gene transfer. Nature 305: 440–443, 1983
Malissen, B., Peele Price, M., Goverman, J. M., McMillan, M., White, J., Kappler, J., Marrack, P., Pierres, A., Pierres, M., and Hood, L.: Gene transfer of H-2 class II genes: Antigen presentation by mouse fibroblasts and hamster B cell lines. Cell 36: 319–327, 1984
Marrack, P., Endres, R., Shimonkevitz, R., Zlotnik, A., Dialynas, D., Fitch, F., and Kappler, J.: The major histocompatibility complex-restricted antigen receptor on T cells. II. Role of the L3T4 product. J. Exp. Med. 158: 1077–1091, 1983
Naquet, P., Pierres, A., and Pierres, M.: Dissection of the poly(Glu60 Ala30 Tyr10) (GAT)-specific T-cell repertoire in H-2I k mice. I. GAT plus self-I-Ak-reactive T-cell clones can recognize alloactivating and/or restriction determinants on nonself Ia molecules. Immunogenetics 18: 475–488, 1983a
Naquet, P., Marchetto, S., and Pierres, M.: Dissection of the poly(Glu60 Ala30 Tyr10) (GAT)-specific T-cell repertoire in H-2I k mice. II. The use of monoclonal antibodies to study the recognition of la antigens by GAT-reactive T cell clones. Immunogenetics 18: 559–574, 1983b
Norcross, M. A., Bentley, D. M., Margulies, D. H., and Germain, R. N.: Membrane Ia expression and antigen-presenting accessory cell function of L cells transfected with class II major histocompatibility complex genes. J. Exp. Med. 160: 1316–1337, 1984
Pierres, A. and Pierres, M.: Fine specificity of a proliferating T-cell clone activated by a conformational determinant of the I-Ek molecule. Immunogenetics 15: 399–412, 1982
Pierres, M., Devaux, C., Dosseto, M., and Marchetto, S.: Clonal analysis of B- and T-cell responses to la antigens. I. Topology of epitope regions on I-Ak and I-Ek molecules analyzed with 35 monoclonal antibodies. Immunogenetics 14: 481–495, 1981
Pierres, M., Goridis, C., and Golstein, P.: Inhibition of murine T cell mediated cytolysis and T cell proliferation by a rat monoclonal antibody immunoprecipitating two lymphoid cell surface polypeptides of 94,000 and 180,000 molecular weight. Eur. J. Immunol. 12: 60–69, 1982a
Pierres, A., Schmitt-Verhulst, A. M., Buferne, M., Golstein, P., and Pierres, M.: Characterization of an Lyt-1+ cytolytic T-cell clone specific for a polymorphic domain of the I-Ak molecule. Scand. J. Immunol. 15: 649–625, 1982b
Pierres, A., Naquet, P., Van Agthoven, A., Bekkhoucha, F., Denizot, F., Mishal, Z., Schmitt-Verhulst, A. M., and Pierres, M.: A rat anti-mouse T4 monoclonal antibody (H129.19) inhibits the proliferation of la-reactive T cell clones and delineate two phenotypically distinct (T4+, Lyt-2,3−and T4−, Lyt-2,3+) subsets among anti-la cytolytic T cell clones. J. lmmunol. 132: 2775–2782, 1984
Prystowsky, M. R., Ely, J. M., Beller, D. I., Eisenberg, L., Goldman, J., Goldman, M., Goldwasser, E., Ihle, J., Quintans, J., Remold, H., Vogel, S. N., and Fitch, F. M.: Alloreactive cloned T cell lines. VI. Multiple lymphokine activities secreted by helper and cytolytic cloned T lymphocytes. J. Immunol. 129: 2337–2343, 1982
Rabourdin-Combe, C. and Mach, B.: Expression of HLA-DR antigens at the surface of mouse L cells cotransfected with cloned human genes. Nature 303: 670–674, 1983
Rosenthal, A. S.: Determinant selection and macrophage function in genetic control of the immune response. Immunol. Rev. 40: 136–152, 1978
Springer, T. A., Scott-Thomson, W., Miller, L. J., Schmalstieg, F. C., and Anderson, D. C.: Inherited deficiency of the Mac-1, LFA-1, p150,95 glycoprotein family and its molecular basis. J. Exp. Med. 160: 1901–1918, 1984
Watts, T. H., Brian, A. A., Kappler, J. W., Marrack, P., and McConnell, H. M.: Antigen presentation by supported planar membrane containing affinity purified I-Ad. Proc. Natl. Acad. Sci. U.S.A. 81: 7564–7568, 1984
Ziegler, H. K. and Unanue, E. R.: Identification of a macrophage antigen-processing event required for Iregion restricted antigen presentation to T lymphocyte. J. Immunol. 127: 1869–1875, 1981
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Naquet, P., Malissen, B., Bekkhoucha, F. et al. L3T4 but not LFA-1 participates in antigen presentation by Ak-positive L-Cell transformants. Immunogenetics 22, 247–256 (1985). https://doi.org/10.1007/BF00404484
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DOI: https://doi.org/10.1007/BF00404484