Induction of cytotoxic T lymphocytes against cervical cancer cell line CaSki using therapeutic dendritic cell vaccine / 肿瘤

ABSTRACT

Objective: To observe the inductive effect of cytotoxic T lymphocytes (CTLs) against human cervical cancer cell line CaSki using therapeutic dendritic cells (DCs) vaccine in vitro . Methods:. Immature mouse DCs were isolated and cultured. The expressions of cell-surface CD40, CD86, major histocompatibility complex (MHC)-. and CD11c in immature DCs were detected by flow cytometry (FCM). Then the immature mouse DCs were infected with recombinant adenoviral vector carrying human papillomavirus (HPV )16 E 6/E 7 (pAd-E6/E7), and the CaSki cell lysate-loaded autologous DCs vaccine was prepared. The expression of green fluorescent protein in pAd-E6/E7-infected immature mouse DCs was observed under a laser scanning confocal microscope, and the expression of E6 protein was detected by Western blotting. DCs vaccine was used to induce specific CTLs, were subsequently co-cultured with CaSki cells. The killing effect of CTLs against CaSki cells was determined using cell counting kit8(CCK8) assay. Results: HPV16 E6/E7-specific DCs vaccine was successfully prepared. CTLs which induced by DCs vaccine exerted a killing effect on CaSki cells. This killing effect was higher in pAd-E6/E7-infected group than those in CaSki cell lysate-loaded group and the untreated control group (P<0.05). Conclusion: Genetically modified DC vaccine can successfully be prepared by infection with pAd-E6/E7, and it has a significant effect on triggering of specific CTLs against CaSki cells.