Design, synthesis and verification of HPV16 and HPV18 multiple antigens peptides.

BACKGROUNDS: Due to lack of immunotherapy of cervical cancer, human papillomavirus (HPV) multiple antigen peptide (MAP) were constructed and verified to explore a new approach for cervical cancer immunotherapy. METHODS: HPV16MAP and HPV18MAP were added to HLA-A2-positive PBMC to verify their immune effects via enzyme-linked immunospot (ELISPOT). Then, cytotoxic T lymphocytes (CTL) were induced and identified in vitro. ELISPOT detected the number of activated CTL stimulated by corresponding antigens. The killing effect of CTL on SiHa and HeLa cells was analyzed by CCK8 and apoptosis assay. The mouse model was subcutaneously injected with polypeptide and the corresponding peptides were added to lymphocytes for ELISPOT. CCK8 and flow cytometry were used to detect the growth of TC1 cells and U14/HPV18E6E7 cells after co-culture with lymphocytes. RESULTS: The number of lymphocyte spotting format counting (SFU) stimulated by adding HPV16MAP and HPV18MAP was significantly higher. HPV16MAP and HPV18MAP could induce specific CTL and effectively kill cervical cancer cells in vitro. The tumors of mice injected with HPV16MAP and HPV18MAP were significantly reduced. The results of CCK8 suggested that HPV18MAP-specific CTL had a significantly greater inhibitory effect on U14/HPV18E6E7 proliferation. And apoptosis rate of HPV16MAP and HPV18MAP-specific CTL was significantly increased. CONCLUSIONS: HPV16 MAP and HPV18MAP have good immune effects and peptide-binding ability in vitro and vivo. They may be potential in cervical cancer immunotherapy. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12967-025-07429-8.