pEGFP-C3-B7.2-MAGE-1融合蛋白诱导特应性抗肿瘤免疫应答的研究  

Specific anti-tumor immune response induced by pEGFP-C3-B7.2-MAGE-1 fusion DNA vaccine

作　　者：艾敏[1] 赵国强[1] 张巧[1] 杨胜利[1] 

机构地区：[1]郑州大学基础医学院,郑州450052

出　　处：《中国免疫学杂志》2007年第10期895-898,共4页Chinese Journal of Immunology

基　　金：河南省自然基金资助项目(No.0411042400)

摘　　要：目的:构建重组表达载体pEGFP-C3-B7.2-MAGE-1,研究人B7.2/MAGE-1基因修饰的食管癌细胞作为瘤苗的抗肿瘤作用。方法:通过RT-PCR扩增B7.2和MAGE-1的cDNA,以pEGFP-C3为载体,构建pEGFP-C3-B7.2-MAGE-1重组载体,通过脂质体转染技术转染人食管癌细胞株EC9706,外周血来源的树突状细胞(DC)负载肿瘤抗原后,与自体T淋巴细胞共培养3天,获得细胞毒性T淋巴细胞(CTL),用四甲基偶氮唑蓝(MTT)法检测CTL对转染和未转染pEGFP-C3-B7.2-MAGE-1的食管癌细胞EC9706的杀伤活性。结果:CTL对转染pEGFP-C3-B7.2-MAGE-1的肿瘤细胞的杀伤活性大于转染pEGFP-C3和未转染细胞的杀伤活性(P<0.05)。结论:成功构建了真核共表达载体pEGFP-C3-B7.2-MAGE-1,人B7.2/MAGE-1基因修饰的EC9706肿瘤细胞瘤苗,可诱导出明显的抗EC9706细胞的免疫效应。Objective:To construct pEGFP-C3-B7.2-MAGE-1 eukaryotic expression plasmids and to study the immunological effects of esophageal cancer cell vaccine modified by human B7.2/MAGE-1 gene.Methods:The B7.2 and MAGE-1 cDNA,which was amplified by RT-PCR, and the eukaryotic expression plasmid pEGFP-C3-B7.2-MAGE-1 was constructed. Human esophageal cancer cell EC9706 was transfected with the vector of pEGFP-C3-B7.2-MAGE-1 using the technique of lipofectamine transfection.The dendritic cells(DCs)from peripheral blood mononuclear cells(PBMC)were loaded with the tumor antigen,and co-cultured with congeneric T cells derived from PBMCs for 3 days to obtain the tumor specific cytotoxicity T lymphocytes(CTL). Methy1 thiazoly1 tetrazolium (MTT) assay was used to detect inhibition effect of CTL on transfected and untransfected EC9706 cells.Results:The CTL had stronger inhibition response against the cancer cells transfected with pEGFP-C3-B7.2-MAGE-1 than to the cancer cells transfected with pEGFP-C3 and the untransfected cancer cells(P<0.05).Conclusion:The tumor vaccine modified by human B7.2-MAGE-1gene can significantly induce response to kill esophageal cancer cells.

关 键 词：B7.2 MAGE-1 真核表达载体 肿瘤免疫 

分 类 号：Q782[生物学—分子生物学]