PADRE/MUC4重组腺病毒转染树突状细胞诱导特异性CTL体外杀伤作用研究

Generation and cytotoxic analysis of PADRE/MUC4 specific CTL elicited by DC infected with recombinant adenovirus in vitro

目的:观察PADRE/MUC4重组腺病毒转染树突状细胞(DC)诱导特异性细胞毒性T细胞(CTL)及体外特异性杀伤作用。方法:pAd-CMV-PADRE/MUC4转染HLA-A2健康志愿者外周血单个核细胞(PBMC)来源的未成熟DC,TNF-α诱导成熟后与自体PBMC混合培养刺激3周,Cr51和Elispot实验检测CTL体外杀伤活性。结果:Cr51和Elispot检测结果显示PADRE/MUC4转染DC可以诱导产生特异性CTL,而pAd-CMV-GFP组和空白对照组不能产生有效特异性CTL。结论:腺病毒载体介导多表位嵌合基因(PADRE/MUC4)转染未成熟DC,在体外可以诱导产生特异性CTL,对表达MUC4肿瘤细胞具有杀伤效应。

Objective：To discuss the cytotoxic activity of CTL,stimulated by adenovirus containing the universal DR-restricted Th helper epitope（PADRE） combined with HLA-A1 and HLA-A2 restricted epitopes from mucin4 gene（PADRE/MUC4） infected DCs. Methods：Recombinant adenovirus with PADRE/MUC4 gene was prepared,DCs of HLA-A2 positive donors were induced from PBMC in vitro. PBMC primed with adenovirus infected DC for 3 weeks,the specific cytotoxic activity of CTL was detected by standard 4 h Cr^51 release assay and Elispot method. Results：Lymphocytes primed with pAd-CMV-PADRE/MUC4 caused potent cytotoxic responses. By contrast,lymphocytes primed with a GFP expressing adenovirus or mock-infected DCs had no cytotoxic effect. Conclusion：Infection of DCs with an adenovirus encoding PADRE combined with epitopes from mucin4 gene may be one possible strategy for immunotherapy of MUC4-expressing tumors.