Previously we have demonstrated that two doses of a cytotoxic T lymphocyte (CTL) epitope-based peptide vaccine of human papillomavirus type 16 (HPV 16) E7 aa 49-57 elicit protection against outgrowth of HPV 16-transformed tumor cells (C3 cells) in B6 mice. Incomplete Freund's adjuvant (IFA), as a carrier, was used to induce this response. To avoid side effects caused by the use of external adjuvants, we have now investigated the effectiveness of highly purified spleen dendritic cells (DC) that efficiently induce primary peptide-specific CTL responses in vitro, as physiological carriers for the HPV 16 E7(49-57) peptide-based vaccine. This is the first report demonstrating that mice immunized once i.v. with syngeneic spleen DCs pulsed with the HPV 16 E7(49-57) peptide in vitro were protected against the outgrowth of C3 tumor cells. In comparison, a single injection of the HPV 16 E7(49-57) peptide in IFA s.c. also resulted in effective induction of tumor-specific immunity in vivo. In both immunization protocols, protective tumor-specific immunity was mediated by CTL that recognized HPV 16 E7(49-57) peptide-pulsed target cells, as well as C3 cells in vitro. Peptide affinity of the CTL induced by both protocols was similar. Thus under the conditions tested, a single injection of spleen DCs pulsed with a CTL epitope-based peptide in vitro elicited tumor-antigen-specific CTL in vivo, which protected mice against a subsequent tumor inoculation. This result indicates that spleen DCs pulsed with a CTL epitope can effectively serve as a tumor-specific vaccine.