The simian virus 40 (SV40) large T antigen was used as an immunogenic vector to express a herpes simplex virus type 1 (HSV-1) glycoprotein B (gB), H-2Kb-restricted cytotoxic T lymphocyte (CTL) recognition epitope corresponding to amino acid residues 498-505. Immunization of naive, C57BL/6 mice with a cell line, B6/350gB, expressing the chimeric T antigen was able to induce the generation of gB498-505-specific CTL in both the lymph nodes and the spleen. Splenic-derived, gB498-505-specific memory CTL (CTLm) were detected in these mice for at least 6 months following immunization at a slightly lower frequency than in those mice immunized with infectious HSV-1. B6/350gB was also able to activate in vitro gB498-505-specific memory CTL obtained from mice previously challenged with HSV. Overall, these findings support the use of a chimeric T antigen as a vector in determining the immunogenic potential of individual CTL epitopes and to assess their potential contribution in inducing a protective immune response in vivo.