Comparison of six cell penetrating peptides with different properties for in vitro and in vivo delivery of HPV16 E7 antigen in therapeutic vaccines.

The high risk human papillomavirus (HPV) types 16 and 18 are globally linked to >50% and 20% of all cervical cancers, respectively. The HPV E7 oncoprotein was determined as a therapeutic vaccine target due to its constitutive expression by HPV-infected cells. The findings demonstrated the efficiency of therapeutic HPV DNA- and protein-based vaccines in preclinical and clinical trials. However, there are limitations for penetration of DNA and protein constructs into the cells without a suitable delivery system. Recently, several cell penetrating peptides (CPPs) have been suggested for delivery of nucleic acids and proteins into cells through covalent or non-covalent fashion. In this study, we determined highly efficient CPPs for the controlled delivery of HPV16 E7 antigen, in vitro and in vivo. Our data indicated the effective delivery of E7 protein by Pep-1, Cady-2, P28 and hPP10, and E7 DNA by MPG and +36 GFP CPPs in HEK-293T cell line at certain ratios. Moreover, immunization with the heterologous MPG + E7 DNA prime/P28 + rE7 protein boost elicited a higher Th1 cellular immune response with a predominant IFN-γ profile and strong Granzyme B secretion than those induced by other groups in a murine tumor model. Indeed, the groups vaccinated with rE7+ P28/rE7+ P28, MPG+ E7 DNA/P28+ rE7, and E7 DNA+ MPG/E7 DNA+ MPG nanovaccines displayed complete protection and remained tumor-free >60 days after treatment. These data suggest P28 and MPG as promising protein and gene delivery systems for development of HPV therapeutic vaccines.