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目的:研究人B7-2瘤苗和负载了肿瘤细胞冻融抗原的DC瘤苗体外联合诱导抗食管癌的免疫作用.方法:应用脂质体转染技术,将融合基因表达载体pEGFP-N3-B7—2转染人食管癌细胞株EC9706.采用密度梯度离心法从脐血中分离单个核细胞(MNC)后,获得单核细胞(Mo).在细胞因子作用下诱导分化,第3d加入EC9706的冻融抗原,共培养4d后获得负载肿瘤抗原的成熟树突状细胞(MDC).将致敏DC与从脐血中分离的T淋巴细胞共培养3d,获得细胞毒T淋巴细胞(CTL);四甲基偶氮唑蓝(MTT)法检测CTL对转染和未转染的EC9706的细胞毒作用.结果:融合基因在食管癌细胞株EC9706的胞膜上定位表达.脐血来源的DC可负载并递呈肿瘤抗原,激活自体T淋巴细胞,诱导肿瘤特异性CTL产生,对转染pEGFP—N3-B7—2的EC9706细胞有显著杀伤作用(F=21.672,P=0.000).结论:人B7—2瘤苗和DC瘤苗体外联合应用,诱导出明显的杀伤食管癌细胞的免疫效应.
OBJECTIVE: To study the immune response of human B7-2 tumor vaccine and DC vaccine loaded with tumor cell freezing-thawing antigen in vitro.Methods: The fusion gene expression vector pEGFP-N3- B7-2 was transfected into human esophageal cancer cell line EC9706.Mononuclear cells (MNCs) were isolated from cord blood by density gradient centrifugation and induced to differentiate under the action of cytokines. The cells were treated with EC9706 Frozen-thawed antigens were co-cultured for 4 days to obtain mature tumor-bearing dendritic cells (MDC) loaded with tumor antigens.To sensitized DCs were co-cultured with T lymphocytes isolated from umbilical cord blood for 3 days to obtain cytotoxic T lymphocytes (CTLs) ; MTT assay was used to detect the cytotoxic effect of CTL on transfected and non-transfected EC9706 cells.Results: The fusion gene was localized on the cell membrane of esophageal carcinoma cell line EC9706.Cell-derived DCs The tumor cells could be loaded and presented with tumor antigens, activating autologous T lymphocytes and inducing tumor-specific CTL production, which markedly killed EC9706 cells transfected with pEGFP-N3-B7-2 (F = 21.672, P = 0.000) Combined application of human B7-2 tumor vaccine and DC vaccine in vitro induced obvious immunity to kill esophageal cancer cells Should.