Tehran University of Medical Sciences Iranian Journal of Allergy, Asthma and Immunology 1735-1502 24 4 2025 06 26 498 507 Maryam Mashhadi Abolghasem Shirazi Department of Molecular Virology, Pasteur Institute of Iran, Tehran, Iran AND Department of Microbiology, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran Seyed Mehdi Sadat Department of Hepatitis, AIDS and Blood Borne Diseases, Pasteur Institute of Iran, Tehran, Iran Nasser Hashemi Goradel Department of Medical Biotechnology, Maragheh University of Medical Sciences, Maragheh, Iran Arash Arashkia Department of Molecular Virology, Pasteur Institute of Iran, Tehran, Iran 2024 09 16 2025 01 04 Despite great efforts in developing peptide-based therapeutic vaccines against human papillomavirus (HPV)-induced cervical cancers, they have failed to elicit strong and sustainable immune responses. Here, we evaluated the vaccine potential of an HPV16 three mutant of E7 (E7GGG) (D21G/C24G/E26G) protein combined with Aldara (topical imiquimod) adjuvant in a TC-1 mouse tumor model. The HPV16-E7GGG, with eliminated transforming properties but retained antigenicity, and E7 wild-type were inserted into pET28, expressed in the E coli system, and purified using Ni-NTA chromatography. The E7GGG and E7 wild-type proteins were combined with Aldara adjuvant and injected into C57BL mice. We determined the ability of HPV16-E7GGG in combination with Aldara adjuvant to induce robust immune responses by IgG total development, IL-4, IL-17, and IFN-γ induction, CTL activity, and inhibit tumor growth in the murine TC-1 model in different immunized groups. The generated recombinant HPV16-E7GGG induced humoral and cellular immune responses in a TH1-mediated pathway, specifically with the (E7GGG) (D21G/C24G/E26G) antigen combined with Aldara, which could be a suitable therapeutic vaccine candidate against HPV. https://ijaai.tums.ac.ir/index.php/ijaai/article/view/4204 https://ijaai.tums.ac.ir/index.php/ijaai/article/download/4204/2192