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

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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.