【摘 要】
：
本研究探讨异种蛋白neu-Fc协同重组人粒/巨噬细胞集落刺激因子(GM-CSF)及卡介苗(BCG)对Th1型和Th2型免疫应答的调节作用.通过基因工程技术将大鼠neu L2-S2功能域与人IgG Fc区
【机 构】
：
军事医学科学院基础医学研究所细胞免疫学实验室,河南大学医学院细胞与分子免疫学重点实验室,新加坡国立大学医院医学部
论文部分内容阅读
本研究探讨异种蛋白neu-Fc协同重组人粒/巨噬细胞集落刺激因子(GM-CSF)及卡介苗(BCG)对Th1型和Th2型免疫应答的调节作用.通过基因工程技术将大鼠neu L2-S2功能域与人IgG Fc区编码序列融合,构建真核表达载体.在CHO细胞中获得稳定表达,应用rProtein A Sepharose Fast Flow亲和层析纯化neu-Fc融合蛋白,应用SDS-PAGE和Western blot鉴定重组蛋白.Ficoll法分离人外周血单个核细胞(PBMNC),MTT法检测融合蛋白对PBMNC的促增殖作用.通过ELISA法检测IL-12和IL-10水平的变化.结果表明:PBMNC经MCF-7细胞上清诱导后,IL-12水平降低(P＜0.05),IL-10水平显著升高(P＜0.01).10 nmol/L neu-Fc对PBMNC具有显著的促增殖作用.与GM-CSF、BCG或者neu-Fc单独作用相比,neu-Fc与GM-CSF、BCG联合作用可显著上调IL-12表达,下调IL-10表达(P＜0.01).结论:neu-Fc具有很好的促增殖作用,GM-CSF和BCG协同neu-Fc能有效诱导Th1型细胞应答.
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