冻融人树突状细胞基因疫苗体外抗肿瘤免疫效应

目的：观察人外周血和人脐血来源的冻融树突状细胞（dendritic cell,DC）基因疫苗的形态、表型及体外诱导的CTL抗肿瘤活性。方法：细胞因子扩增人外周血和脐血DC,分别将EBV-LMP2、HPV16E6基因转染2种来源的冻融DC制备疫苗。动态形态学观察和流式细胞术检测疫苗表面分子表达,体外诱导并测定CTL活性。结果：人外周血和脐血冻融DC疫苗均高表达CD80、CD86和CD83,低表达CD14,高表达CD1a,体外均能诱导高效的CTL活性（P=0.001）,并与新鲜DC疫苗差异无统计学意义,P=0.138。结论：负载肿瘤相关病毒抗原基因的冻融DC疫苗保持了功能成熟DC的形态特征,且能诱导高效的特异性抗肿瘤免疫应答。

Specific antitumor immunological effect of cytotoxic T cell induced by human cryopreserved dendritic cell transfected with viral antigen gene

OBJECTIVE:To observe the biological characteristics of cryopreserved dendritic cell (DC) vaccine derived from human peripheral blood and cord blood and transfected with viral antigen gene, and study its antitumor immunological effects in vitro. METHODS: DC-enriched populations were prepared from human peripheral blood and cord blood in vitro cultures with different cytokines, and then cryopreserved in liquid nitrogen. Two different viral antigen genes were transfected into two different cryopreserved DCs respectively. The morphology of DC was observed dynamically, and the phenotypes of DC vaccine were analyzed by flow cytometry. MTT assay was applied to detect the cytotoxicity activity of CTL in vitro. RESULTS: Cryopreserved DC vaccine had typical morphologic characteristics, highly expressed CD80, CD86 and CD83, and showed individually low CD14 and high CD1a expressions. The cytotoxicity activities of CTL induced by two different derived cryopreserved DC vaccines were both significantly higher than those of the control groups, P=0.001. There was no significant difference between the cryopreserved DC vaccine and fresh DC vaccine, P=0.138. CONCLUSION: Cryopreserved DC transfected viral antigen gene could sustain the morphology and phenotype of mature DC, and could induce effectively specific potent CTL activity in vitro.