Webb Nicholas E, Lee Benhur
Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA, USA.
Department of Microbiology, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, #1124, New York, NY, 10029, USA.
Methods Mol Biol. 2016;1354:3-20. doi: 10.1007/978-1-4939-3046-3_1.
Entry of HIV-1 into target cells involves the interaction of the HIV envelope (Env) with both a primary receptor (CD4) and a coreceptor (CXCR4 or CCR5). The relative efficiency with which a particular Env uses these receptors is a major component of cellular tropism in the context of entry and is related to a variety of pathological Env phenotypes (Chikere et al. Virology 435:81-91, 2013). The protocols outlined in this chapter describe the use of the Affinofile system, a 293-based dual-inducible cell line that expresses up to 25 distinct combinations of CD4 and CCR5, as well as the associated Viral Entry Receptor Sensitivity Assay (VERSA) metrics used to summarize the CD4/CCR5-dependent infectivity results. This system allows for high-resolution profiling of CD4 and CCR5 usage efficiency in the context of unique viral phenotypes.
HIV-1进入靶细胞涉及HIV包膜（Env）与主要受体（CD4）和共受体（CXCR4或CCR5）的相互作用。特定Env利用这些受体的相对效率是进入过程中细胞嗜性的主要组成部分，并且与多种病理性Env表型相关（Chikere等人，《病毒学》435:81-91，2013年）。本章概述的方案描述了Affinofile系统的使用，这是一种基于293的双诱导细胞系，可表达多达25种不同组合的CD4和CCR5，以及用于总结CD4/CCR5依赖性感染性结果的相关病毒进入受体敏感性分析（VERSA）指标。该系统允许在独特病毒表型的背景下对CD4和CCR5的使用效率进行高分辨率分析。
