Background: Cervical cancer, the second leading cause of cancer mortality in women worldwide, results from an infection with a subset of human papillomavirus (HPV), HPV-16 being the most prevalent type. The available prophylactic vaccines are an effective strategy to prevent this cancer but decades will be needed before it may be eradicated. Therapeutic vaccines are necessary to eliminate infected cells and associated lesions in women who have no benefit from prophylactic vaccination. Until now, therapeutic vaccines only showed poor clinical results, possibly linked to an inefficient targeting of protective immune responses in the genital mucosa. Methods: Mice were immunized via mucosal or parenteral routes with a synthetic HPV16 E71-98 polypeptide vaccine administered with different adjuvants. E7-specific CD8 T cell responses were evaluated by IFN-g ELISPOT in the blood, draining lymph nodes and in the genital mucosa. Results: Parenteral vaccination with CpG and HLT adjuvants induced high E7-specific responses in the blood, whereas combination of Resiquimod, HLT and CpG was the best combination after an aerosol immunization, although this response was 5-fold lower than after the parenteral vaccination. The amplitude and kinetics of E7-responses were also measured in draining lymph nodes and more importantly in the genital tissue itself. Despite the lower E7-specific response in the periphery after the aerosol immunization, the responses measured in the genital mucosa after both types of vaccination were similar, suggesting a preferential genital homing after aerosol vaccination. Interestingly, there was no correlation between the responses measured in the periphery with those measured in the genital mucosa, highlighting the necessity to determine the immune responses in the mucosa where the tumor reside. Finally, we have evidences that additive applications of topical immunomodulators locally enhanced the therapeutic properties of the E71-98 polypeptide vaccine. Conclusion: Our results suggest that combination of the adjuvanted E71-98 peptide with topical immunomodulators could be a potent therapeutic vaccine against HPV-16 and cervical cancer.