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
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国家自然科学基金资助项目(30500492和30471691)


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

    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-

    Abstract:

    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 Cr51 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.

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吴峻立,卫积书,孟 凯,高文涛,陈建敏,张静静,朱 毅,苗 毅. 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[J].南京医科大学学报(自然科学版),2007,(12):1349-1352

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  • 收稿日期:2007-10-18
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