HIV-1临床株SC42无感染性细胞-细胞融合模型的构建  

Construction of a non-infectious cell-cell fusion model of HIV-1 clinical strain SC42

作　　者：石哲芳 魏雪玲 刘奇 SHI Zhe-fang;WEI Xue-ling;LIU Qi(Integrated Lab of Pathogenic Biology,Pre-clinical College,Dali University,Dali 671000,Yunnan,China)

机构地区：[1]大理大学基础医学院病原生物学综合实验室,云南大理671000

出　　处：《中国病原生物学杂志》2022年第8期892-895,900,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.81703573,81660337);大理大学创新团队项目(No.ZKLX2019105)。

摘　　要：目的构建一种无感染性、可视化,用于检测HIV-1进入抑制剂在临床株SC42上的抑制活性的HIV细胞-细胞融合模型。方法将HIV-1 SC42 env基因插入质粒pAAV-IRES-EGFP,获得可同时且分别表达HIV-1 SC42 env及GFP的真核表达质粒,经过转染293T细胞,使之同时表达这两种蛋白(293T/SC42/EGFP),作为模拟HIV病毒的效应细胞与靶细胞TZM-b1共同培养,观察细胞融合情况及合胞体的形成。使用经典HIV-1进入抑制剂C34和T1144检测该细胞融合模型的效果。结果效应细胞293T/SC42/EGFP和靶细胞TZM-b1细胞混合2 h后,在荧光显微镜下可见明显的细胞融合现象;24 h后可形成合胞体。HIV-1进入抑制剂C34、T1144能够抑制细胞融合,其IC分别为(600±6.22)nmol/L和(44±5.88)nmol/L。结论成功构建了HIV-1进入抑制剂药物检测筛选模型。该模型的构建无需在P3生物安全实验室完成,操作简单、方便且无感染性。Objective To construct a non-infectious,visualized HIV cell-cell fusion model for testing the inhibitory activity of HIV-1 entry inhibitors on clinical strain SC42.Methods By inserting the HIV-1 SC42 env gene into the plasmid pAAV-IRES-EGFP,a eukaryotic expression plasmid that can simultaneously and separately express HIV-1 SC42 env and GFP was obtained,and 293 T cells were transfected to express these two proteins simultaneously(293 T/SC42/EGFP),as effector cells mimicking HIV virus,co-cultured with target cells TZM-b1,and observed cell fusion and the formation of syncytia.The effect of this cell fusion model was tested using the classical HIV-1 entry inhibitors C34 and T1144.Results After mixing effector cells 293 T/SC42/EGFP and target cells TZM-b1 cells for 2 hours,obvious cell fusion phenomenon can be seen under the fluorescence microscope;after 24 hours,syncytia can be formed.HIV-1 entry inhibitors C34 and T1144 can inhibit cell fusion with ICof 600±6.22 nmol/L and 44±5.88 nmol/L,respectively.Conclusion A screening model for HIV-1 entry inhibitor drug detection was successfully constructed.The construction of this model does not need to be completed in the P3 biosafety laboratory,and the operation is simple,convenient and non-infectious.

关 键 词：HIV-1 SC42 ENV 进入抑制剂 细胞-细胞融合 

分 类 号：R373.9[医药卫生—病原生物学]