CYTOTOXIC T-LYMPHOCYTE ESCAPE VARIANTS, INDUCED MUTATIONS, AND SYNTHETIC PEPTIDES DEFINE A DOMINANT H-2K(B)-RESTRICTED DETERMINANT IN SIMIAN-VIRUS-40 TUMOR-ANTIGEN

Immunization of C57BL/6 mice with syngeneic cells transformed by simia n virus 40 large T antigen (SV40 T ag) induces the generation of T ant igen-specific cytotoxic T lymphocytes (CTL) which are restricted by th e major histocompatibility class I antigens H-2D(b) and H-2K(b). Previ ous studies have shown that the H-2D(b)-restricted CTL response is dir ected to at least three distinct epitopes (I, II/III, and V) in the SV 40 T antigen which have been precisely mapped using deletion mutagenes is and overlapping synthetic peptides. Although in vivo the CTL respon se to SV40 T antigen is dominated by the H-2K(b) class I antigen, the precise location of the H-2K(b)-restricted epitope(s) was not known, a nd whether there was multiplicity of H-2K(b)-restricted epitopes remai ned unclear. In this study, we have defined the minimal recognition ep itope for the SV40-specific H-2K(b)-restricted CTL clone Y-4 as T anti gen residues 404-411 by using T antigen deletion and point mutants and synthetic peptides. DNA sequence analysis of the region encoding resi dues 404-411 from the T antigens expressed in three independently isol ated CTL clone Y-4 escape variants identified inactivating mutations c apable of abrogating CTL recognition. Estimation of CTL precursor (CTL p) frequencies by limiting dilution analysis revealed that CTLp specif ic for epitope IV represent a large percentage of the total CTL respon se elicited by the intact T antigen in H-2(b) mice. Immunization of B6 mice with cells expressing a T antigen derivative deleted of residues 404-411 revealed that site IV represents the only immunodominant H-2K (b)-restricted epitope within T antigen. (C) 1995 Academic Press, Inc.