title: Quantifying CD4/CCR5 Usage Efficiency of HIV-1 Env Using the Affinofile System.
abstract: 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.
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA, USA. hydrazine@ucla.edu.
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Webb
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, #1124, New York, NY, 10029, USA. benhur.lee@mssm.edu.
authors.forename: Benhur
authors.initials: B
authors.lastname: Lee
mesh_terms: D000818:Animals; D015704:CD4 Antigens / Q000032:analysis / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D005434:Flow Cytometry / Q000379:methods; D015658:HIV Infections / Q000378:metabolism* / Q000821:virology; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D019713:Receptors, CCR5 / Q000032:analysis / Q000378:metabolism*; D053586:Virus Internalization; D054299:env Gene Products, Human Immunodeficiency Virus / Q000032:analysis / Q000378:metabolism*
chemical_list: D015704:CD4 Antigens; D019713:Receptors, CCR5; D054299:env Gene Products, Human Immunodeficiency Virus
keywords: Affinofile; CCR5; CD4; Entry efficiency; Env; HIV; Receptor usage efficiency; Virus entry