Previously, we have shown that immunization with human papillomavirus (HPV) type 16‐derived cytotoxic T lymphocyte (CTL) epitope E7 49–57 (RAHYNIVTF) renders C57BL/6 mice insensitive to tumors formed by HPV16‐transformed cells. In this study, we provide evidence that E7 49–57 is expressed as a subdominant CTL epitope on HPV16‐transformed C57BL/6 cells. Using acid peptide elution, it is shown that HPV16‐transformed cells express another CTL epitope, besides E7 49‐57, which appears to be dominant. We demonstrate that a CTL line raised against the subdominant CTL epitope, offered as synthetic peptide E7 49–57, eradicates established HPV16‐induced tumors in mice. Our data show that synthetic peptide‐induced CTL can be applied successfully in vivo against (virus‐induced) tumor, and emphasize that subdominant CTL epitopes are useful targets for immunotherapy. Furthermore, it is illustrated for the first time that HPV16‐specific CTL interfere directly with HPV16‐induced tumors.