Restriction fine specificity of long-term, hapten-specific cytotoxic T cell clones: analysis with H-2Kbm-mutant mice and H-2Kb-specific monoclonal antibodies

Abstract

The cell-mediated cytotoxic response against autologous cells modified with the sulfhydryl reagent I-AED [N-iodo-acetyl-N-(5-sulfonic-1-naphthyl) ethylene diamine] has been described by Levy, R. B., Shearer, G. M., Richardson, J. C. and Henkart, P. A., [J. Immunol. 1981. 127: 523.]. We have established two H-2Db- and eight H-2Kb-restricted, AED-specific long-term cytotoxic T lymphocyte (CTL) clones of C57BL/10 origin. The growth of these clones has been dependent upon presence of both antigen and interleukin 2. Cytotoxicity and proliferation analysis of AED-specific Kb-restricted CTL clones with target and stimulator cells from Kbm-mutant mice demonstrated two categories of clones (A and B) based on different reactivity patterns against hapten-modified Kbm-mutant cells. AED-modified target cells of bm5, bm6 and bm9 origin were lysed by type A clones but not by type B clones, in contrast to AED-modified bm3, bm8 and bm11 target cells which were lysed by type B but not by type A clones. None of the clones lysed AED-modified bm1 target cells, but all of them lysed AED-modified bm4 and bm10 target cells. The restriction fine specificities for both cytolytic and proliferative activities of the analyzed clones were identical. A panel of monoclonal antibodies (mAb) directed against the Kb molecule was used to inhibit lysis of AED-modified target cells by CTL clones. mAb which recognize a cluster of allodeterminants located in the second external domain (C1 domain) of the Kb molecule [Hämmerling, G. J., Rüsch, E., Tada, N., Kimura, S. and Hämmerling, U., Proc. Natl. Acad. Sci. USA 1982. 79: 4737] only inhibited type B clones, whereas inhibition of both type of clones was observed with mAb specific for allodeterminants clustered in the N-terminal region of Kb. These findings suggest that different regions on the Kb molecule serve as self determinants for H-2-restricted cytotoxicity. We also demonstrate that allodeterminants recognized by mAb and self determinants involved in H-2 restriction need not be identical. Our data also support the notion that covalent AED modification of H-2 molecules is not necessary to generate the self plus X antigen for CTL recognition.