转染融合基因的DC抗肿瘤免疫效应
Specific Anti-tumor Immunological Effect by Dendritic Cell Transfected with GM-ΔSur Gene
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摘要: 目的 研究转染融合基因（GM-CSF-survivin GM-ΔSur）的树突状细胞（DC）在体外诱导高效而特异的抗肿瘤免疫效应 。方法 用jetPEI^TM-Macrophage转染体系,将构建的GM-ΔSur融合基因转染入DC,用流式细胞仪检测DC的表面分子 HLA-DR、CD83、CD80、CD86表达的高低;用LDH法测定转染融合基因的DC诱导特异性细胞毒性T淋巴细胞（CTLs）杀伤肿瘤细胞（HT-29和OVCAR-3）的能力。结果 转染融合基因的DC细胞中可检测到GM-ΔSur融合蛋白的表达;DC表面高表达HLA-DR、CD83、CD80、CD86;PHA/rhIL-2长期培养（21d）的T细胞CD8^＋比例明显增加;转染融合基因的DC 对HT-29肿瘤细胞的杀伤率显著高于未修饰的DC的杀伤率。结论 GM-ΔSur基因转染修饰的DC能选择性诱导MHC-I类分子限制的CTL的特异性,显著提高DC的抗原提呈功能和诱导高效而特异的抗肿瘤免疫效应。Abstract: Objective To study the specific anti-tumor immunological effects induced by dendritic cells transfected with GM-CSF gene and survivin gene(GM-ΔSur) in vitro. Methods pcDNA 3.1 plasmid containing GM-ΔSur fusion gene was constructed and was transfected into dendritic cells by jetPEITM-Mcrophage transfected kit; cellular surface phenotype such as CD1a, CD80, CD86, CD83 were detected by FACS; The specific cytotoxicity of induced human cytotoxic T lymphocyte (CTLs) to tumor cells was detected with LDH assay. Res...
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