目的运用表达人乳头瘤病毒16型(HPV16)E7蛋白的肿瘤细胞系B16-HPV16E7,探讨pcd-HPV16mE7-HSP70融合DNA疫苗抗肿瘤的可能机制。方法运用流式细胞术检测IFN-γ对B16-HPV16E7细胞表面Fas分子表达的影响。通过ELISA和流式细胞术检测疫苗免疫后小鼠脾淋巴细胞IFN-γ的分泌和FasL的表达情况。运用体外抗体阻断实验检测Fas-FasL途径对pcd-HPV16mE7-HSP70DNA疫苗抗肿瘤作用的影响。结果 pcd-HPV16mE7-HSP70DNA疫苗可以激活机体的免疫系统,上调细胞因子IFN-γ和E7特异性FasL的表达。IFN-γ可以激活B16-HPV16E7肿瘤细胞表面的Fas分子的表达。拮抗FasL后,疫苗对B16-HPV16E7肿瘤细胞的杀伤作用明显减弱。结论 Fas-FasL途径是pcd-HPV16mE7-HSP70DNA疫苗抗肿瘤作用的重要机制之一。
Objective B16-HPV16E7 tumor cells(E7-expression),B16-HPV16E7,were employed to explore the antitumor mechanisms of pcd-HPV16m E7-HSP70 DNA vaccine.Methods Flow cytometry was used to assess the effect of TNF-g on Fas expression on the surface of B16-HPV16E7 cells.IFN-γ and FasL levels from lymphocytes after pcd-HPV16m E7-HSP70 vaccination were measured by ELISA and flow cytometry.CTL assay was used to evaluate the role of Fas-FasL pathway on the anti-tumor ability of pcd-HPV16m E7-HSP70 vaccine.Results IFN-γ stimulation significantly increased Fas expression in B16-HPV16E7 cell line.pcd-HPV16mE7-HSP70 DNA vaccination significantly increased E7-specific lymphocytes and IFN-γ secretion in supernatants of cultured mouse spleen′s lymphocytes.In addition,the specific cytolytic activities of E7-specific lymphocytes against the tumor cells were partly inhibited by anti-FasL antibody pretreatment.Conclusion Pcd HPV16m E7-HSP70 DNA vaccine played an important anti-tumor role through Fas-FasL pathway.