抗人CCR5多克隆抗体体外阻断HIV-1假病毒的感染研究及方案汇总.docxVIP

抗人CCR5多克隆抗体体外阻断HIV-1假病毒的感染研究程林，唐娴，赵方，何美云，王辉（深圳市第三人民医院，广东深圳518112）摘要：目的制备鼠抗人CC型趋化因子受体5（CC chemokine receptor 5，CCR5）特异性HIV-1中和抗体。方法用稳定表达人CCR5的CHO-R5细胞腹腔免疫BALB/c小鼠；流式细胞术的方法分析免疫血清对GHOST-R5细胞的结合；纯化免疫血清总IgG抗体，HIV-1假病毒中和试验分析血清总IgG的中和活性。结果流式细胞术的结果显示，CHO-R5细胞免疫组小鼠血清对GHOST-R5结合的平均荧光强度显著高于对照组（P0.001）；HIV-1假病毒中和试验的结果发现，纯化的CHO-R5细胞组小鼠免疫血清总IgG，可抑制HIV-1假病毒在体外感染GHOST-R5细胞，抑制作用具有剂量依赖性，最高抑制率达到92%。结论成功诱导出具有中和HIV-1活性的鼠抗人CCR5的多克隆抗体。关键词：CC型趋化因子受体5；中和抗体；艾滋病病毒1型中图分类号：R 373.9 文献标志码：A文章编号：1672-5662（2015）11-0000-00Polyclonalantibodies to human CCR5 block the infection of HIV-1 pseudotyped virus in vitroCHENG Lin，TANG Xian，ZHAO Fang，et al.（Shenzhen Third People’s Hospital, Shenzhen 518112, Guangdong China）Abstract:ObjectiveToprepareCCR5-specific HIV-1 neutralizing antibodies.MethodsBALB/c mice were intraperitoneally immunized with human CCR5 stable-expressing CHO-R5 cells；The binding of sera form immunized mice with GHOST-R5 cells was analyzed by flow cytometry;the neutralizing activity of purified total IgG from immunized mice to HIV-1 infection was determined by HIV-1env-pseudotyped virusneutralizing assay.ResultsThe mean fluorescence intensity of the binding of GHOST-R5 cells with sera from mice of CHO-R5 cells immunized group wassignificantly higher than that of the control group (P 0.001) by flow cytometry; and the neutralizing assay shown that, total IgG purified from mice of CHO-R5 cells immunized group effectively inhibited the infection of GHOST-R5 cells by HIV-1 pseudotyped virus in a dose-dependent manner, with the highest inhibition ratio of 92%. ConclusionsSuccessfully induced mouse anti human CCR5polyclonal antibodies with HIV-1neutralizing activity.Keywords：CCR5；Neutralizing antibody；HIV-1近年来，全球新发感染艾滋病病毒1型（human immunodeficiency virus type-1，HIV-1），人数和艾滋病死亡人数总体呈现下降的趋势。中国是疫情较为严重的国家之一，艾滋病仍是面临的重大公共卫生问题[1]。鼠抗人CC型趋化因子受体5（CC chemokine receptor 5，CCR5）作为HIV-1主要的辅助受体，在病毒感染和传播过程中起关键作用[2]。本研究以表达人CCR5的CHO-R5细胞为免疫原，成功在小鼠体内诱导出具有中和活性的CCR5多克隆抗体，为开发靶向CCR5的HIV-1中和抗体奠定基础。1材料方法收稿日期：2015-05