以AFP为靶点的肝癌树突状细胞免疫治疗的实验研究

目的探讨AFP作为肝癌基因治疗和免疫治疗靶点的可行性以及AFP基因转染的树突状细胞（AFP-DC）疫苗对表达AFP肝细胞癌的免疫治疗作用.方法构建AFP-cDNA 真核表达载体,体外转染DC,制备AFP-DC瘤苗,诱导针对表达AFP的肝癌细胞株HepG2的特异性免疫反应,MTT法检测效应细胞对靶细胞的杀伤率.结果 AFP-DC瘤苗能够分泌AFP抗原,培养上清中AFP含量为0.8805 IU/ml,与空载体组和空白对照组相比,差异显著（P〈0.05）.免疫荧光检测可见AFP-DC胞浆及胞膜有AFP抗原分子表达;活化的CTL能够对表达AFP肝癌细胞起特异性杀伤作用,杀伤效率可达84.05%,与空载体组和空白对照组比较,差异显著（P〈0.05）.结论 AFP作为靶点治疗肝癌具有可行性,AFP可作为肝癌靶向治疗的新的突破点;AFP-DC疫苗可以作为表达AFP肝癌的一种免疫治疗手段,为DC瘤苗的临床应用奠定了基础.

Immunotherapy of hepatocellular carcinoma with dendritic cells vaccine targeting to alpha-fetoprotein

Objective To explore the feasibility of AFP being target to hepatocellular carcinoma and immunotherapeutic activity of AFP gene priming dendritic cell vaccine to hepatocellular carcinoma cells expressing AFP antigen molecule.Methods AFP-cDNA eukaryotic expression vector was constructed and transfected into immature dendritic cells induced from peripheral blood mononuclear cells with liposome.The specific CTL was induced ex vivo with the generated AFP-DC vaccine and detected its targeting cytotoxic functions to hepatocellular carcinoma cells expressing AFP.Results The full length AFP-cDNA cloned to the eukaryotic vector successfully detected through electrophoresis,AFP-DC could secrete AFP antigen molecule,there was 0.8805 IU/ml AFP antigen detected in the supernatant of AFP-DC,compared with the control groups,the differences were significant(P<0.05).It also could be detected through immunofluorescence staining.AFP-DC activated CTL presented the specific killer ability to the hepatocellular carcinoma cells expressing AFP,the killer ability rate was 84.05%,compared with the control groups,the differences were significant(P<0.05).Conclusion AFPDC vaccine may be a tool of the hepatocellular carcinoma immunotherapy,it will be foundation of future clinical use of DC vaccine.AFP as a tumor-associated antigen,may be a target to hepatocellular carcinoma expressing AFP.