Kramer Susanne, Buontempo Peter, Agrawal Sony, Ralston Robert
Merck Research Laboratories, Department of Infectious Diseases, Kenilworth, NJ, USA.
J Biomol Screen. 2011 Jul;16(6):668-75. doi: 10.1177/1087057111403480. Epub 2011 Apr 7.
Infection of certain cell types by HIV results in formation of syncytia. This process can be blocked by antibodies or compounds that prevent interaction of viral envelope protein with host cell receptors. Here the authors describe an automated imaging-based assay for inhibitors of cell-cell fusion mediated by interaction of HIV gp120 with CXCR4 coreceptor. The assay quantifies syncytia formation between U87MG astrocytoma cells constitutively expressing CD4/CXCR4 and morphologically distinct Jurkat T lymphoma cells inducibly expressing HIV env. Each cell type was differentially labeled with vital dyes. Fusion was quantified by measuring size, shape, and color of Jurkat cells and Jurkat-harboring cell syncytia. Dose-response experiments with reference inhibitors AMD 3100 and KRH-1636 yielded potencies consistent with those obtained using standard antiviral assays. This assay complements virus-based infectivity assays for identification of inhibitors of membrane fusion events triggered by interaction of HIV gp120 with host CXCR4.
某些细胞类型被HIV感染会导致多核巨细胞的形成。这一过程可被阻止病毒包膜蛋白与宿主细胞受体相互作用的抗体或化合物阻断。本文作者描述了一种基于自动成像的检测方法，用于检测由HIV gp120与CXCR4共受体相互作用介导的细胞间融合抑制剂。该检测方法可量化组成性表达CD4/CXCR4的U87MG星形细胞瘤细胞与可诱导表达HIV env的形态不同的Jurkat T淋巴瘤细胞之间的多核巨细胞形成。每种细胞类型都用活性染料进行了差异标记。通过测量Jurkat细胞和含有Jurkat细胞的多核巨细胞的大小、形状和颜色来量化融合。使用参考抑制剂AMD 3100和KRH-1636进行的剂量反应实验得到的效力与使用标准抗病毒检测方法得到的结果一致。该检测方法补充了基于病毒的感染性检测方法，用于鉴定由HIV gp120与宿主CXCR4相互作用引发的膜融合事件的抑制剂。
