Cytotoxic T Lymphocytes Elicited by Dendritic Cell-Targeted Delivery of Human Papillomavirus Type-16 E6/E7 Fusion Gene Exert Lethal Effects on CaSki Cells

Abstract

Human papillomavirus (HPV) is the primary etiologic agent of cervical cancer. Consideration of safety andnon human leukocyte antigen restriction, protein vaccine has become the most likely form of HPV therapeuticvaccine, although none have so far been reported as effective. Since tumor cells consistently express the twoproteins E6 and E7, most therapeutic vaccines target one or both of them. In this study, we fabricated DCvaccines by transducing replication-defective recombinant adenoviruses expressing E6/E7 fusion gene ofHPV-16, to investigate the lethal effects of specific cytotoxic T lymphocytes (CTL) against CaSki cells in vitro.Mouse immature dendritic cells (DC) were generated from bone marrow, and transfected with pAd-E6/E7 toprepare a DC vaccine and to induce specific CTL. The surface expression of CD40, CD68, MHC II and CD11cwas assessed by flow cytometry (FCM), and the lethal effects of CTL against CaSki cells were determined byDAPI, FCM and CCK-8 methods. Immature mouse DC was successfully transfected by pAd-E6/E7 in vitro,and the transfecting efficiency was 40%-50%. A DC vaccine was successfully prepared and was used to inducespecific CTL. Experimental results showed that the percentage of apoptosis and killing rate of CaSki cells weresignificantly increased by coculturing with the specific CTL (p <0.05). These results illustrated that a DC vaccinemodified by HPV-16 E6/E7 gene can induce apoptosis of CaSki cells by inducing CTL, which may be used as anew strategy for biological treatment of cervical cancer.