重组腺病毒介导白细胞介素-18和甲胎蛋白基因修饰的树突状细胞体外诱导抗肝癌免疫活性的研究

目的 探讨甲胎蛋白(AFP)、白细胞介素(IL)-18基冈修饰树突状细胞(DC)较以往AFP基因修饰DC作为肝癌免疫治疗瘤苗是否更具优势.方法 分离外周血单个核细胞定向诱导为DC,以Ad-IL-18和Ad-AFP共感染DC,感染后细胞通过噻唑蓝(MTT)比色法检测其激活的T细胞对HepG2的杀伤活性.结果 检测结果显示目的 基因能够表达于DC细胞中.流氏细胞仪(FCM)结果显示共感染的DC细胞均高表达CD1a(73.4%)、CD11c(84.3%)、CD80(89.5%)、CD86(87.9%)和HLA-DR(91.7%).CTL结果显示IL-18/AFP-DC-T对HepG2的杀伤率(67.49±3.24)%明显高于对SMMC7721细胞(27.32±1.75)%和K562细胞(17.31±1.56)%的杀伤率,另外共感染IL-18/AFP-DC-T组对HepG2的杀伤率与AFP-DC-T、IL-18-DC-T组比较,其差异有统计学意义.结论 AFP和IL-18基因修饰DC,能够在体外诱导特异性细胞毒性T淋巴细胞(CTL)效应,而_且对HepG2细胞杀伤率大于AFP转染DC组.

Alpha-fetoprotein and IL-18 gene-modified dendritic cells-induced CTL response against hepatocellular carcinoma cells

Objective To investigate the immune functions of dentritic cells (DCs) transduced with Ad-IL-18, Ad-AFP in vitro, compare their antiviral and antitumor effects with those of Ad-IL-18/AdAFP DCs and Ad-AFP DCs.Methods Peripheral blood mononuclear cells (PBMC) from HCC patients were isolated by blood separator.PBMC were cultured in vitro for one week to induce DCs.After transfection with Ad-IL-18 and Ad-AFP (IL-18/AFP-DC) ,the stimulated lymphocytes were tested by cytotoxicity capacity assay.Results The results showed that DCs could infect and express relevant genes.The expression of CD11c (84.3%) ,CD1a (73.4%) ,CD80 (89.5%) ,CD86 (87.9%) and HLA-DR (91.7%) on adenovirus transduced DCs were up-regulated.The cytotoxicity mediated by IL-18/AFP-DC-T was high-er than that mediated by DC-T and T,and that mediated by IL-18/AFP-DC-T was higher than that mediated by IL-18-DC-T and AFP-DC-T.Conclusion DCs co-infected with these vectors can induce antitumor CTL response in vitro.And Ad-AFP and Ad-IL-18 transfected DCs could induce stronger CTL response against HepG2 than AFP gene-transfected DCs.