Three HSV type 1 (HSV-1) and HSV type 2 (HSV-2) common ("HSV-type common") and three HSV-1 specific CTL clones, which were CD3+, CD4+, CD8-, 4B4+, and 2H4-, were established. These clones proliferated in response to stimulation with HSV in the presence of autologous APC. The HSV type specificity of the proliferative response was identical with that of the cytotoxic activity of the clones. The cytotoxic activity and the proliferative response were both inhibited by addition of anti-HLA-DR mAb to the culture. After culture of these CTL clones with autologous B cells and macrophages followed by HSV Ag stimulation, anti-HSV antibody was detected in the culture supernatant. The HSV type specificity of the helper function for antibody production was identical with that of the cytotoxicity, i.e., HSV-type common clones, upon stimulation with either HSV-1, or HSV-2, and HSV-1-specific clones, upon stimulation with HSV-1 but not with HSV-2, showed helper activity for anti-HSV antibody production by autologous B cells. Moreover, it was found that these clones produced humoral factors which help autologous B cells to produce antibody. The helper factors were produced by T cell clones in an HSV-type-specific manner. These data suggest that some CD4+ T cells can simultaneously manifest both specific cytotoxicity and helper activity for Ag-specific antibody production by B cells, and that these multifunctional T cells might play an important role in protection against viral infection.