Denolly Solène, Cosset François-Loïc, Freitas Natalia
CIRI-International Center for Infectiology Research, Team EVIR, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR5308, Ecole Normale Supérieure de Lyon, Univ Lyon, Lyon, France.
Methods Mol Biol. 2019;1911:219-234. doi: 10.1007/978-1-4939-8976-8_15.
The membrane fusion properties of HCV envelope glycoproteins can be evaluated using several assays. Fusion assays generally require contacts between glycoproteins expressed on a donor membrane, such as those from a cell or a viral particle, and an acceptor membrane that may or may not express cognate viral receptor, such as those from an indicator cell or a liposome. In this chapter, we describe three well-established methods in the field that use either cell surface expression of glycoproteins, HCV pseudoparticles (HCVpp), or cell culture-grown HCV (HCVcc) particles for donor membrane and cells or liposomes as acceptor membrane in which specific components can be included to monitor and quantify fusion. We provide details of cell-cell fusion assay, virus-liposome fusion assay, and finally virus-plasma membrane fusion assay. We also describe inhibitors that can block HCV envelope membrane fusion.
丙型肝炎病毒包膜糖蛋白的膜融合特性可通过多种检测方法进行评估。融合检测通常需要供体膜(如来自细胞或病毒颗粒的膜)上表达的糖蛋白与受体膜(可能表达也可能不表达同源病毒受体,如来自指示细胞或脂质体的膜)之间发生接触。在本章中,我们描述了该领域三种成熟的方法,这些方法分别使用糖蛋白的细胞表面表达、丙型肝炎病毒假颗粒(HCVpp)或细胞培养产生的丙型肝炎病毒(HCVcc)颗粒作为供体膜,以细胞或脂质体作为受体膜,其中可包含特定成分来监测和量化融合。我们详细介绍了细胞 - 细胞融合检测、病毒 - 脂质体融合检测,最后是病毒 - 质膜融合检测。我们还描述了可阻断丙型肝炎病毒包膜膜融合的抑制剂。