title: A combination HIV reporter virus system for measuring post-entry event efficiency and viral outcome in primary CD4+ T cell subsets.
abstract: Fusion between the viral membrane of human immunodeficiency virus (HIV) and the host cell marks the end of the HIV entry process and the beginning of a series of post-entry events including uncoating, reverse transcription, integration, and viral gene expression. The efficiency of post-entry events can be modulated by cellular factors including viral restriction factors and can lead to several distinct outcomes: productive, latent, or abortive infection. Understanding host and viral proteins impacting post-entry event efficiency and viral outcome is critical for strategies to reduce HIV infectivity and to optimize transduction of HIV-based gene therapy vectors. Here, we report a combination reporter virus system measuring both membrane fusion and viral promoter-driven gene expression. This system enables precise determination of unstimulated primary CD4+ T cell subsets targeted by HIV, the efficiency of post-entry viral events, and viral outcome and is compatible with high-throughput screening and cell-sorting methods.
authors.affiliation: Center for Proteomics and Bioinformatics, School of Medicine, Case Western Reserve University, 919 BRB 10900 Euclid Ave, Cleveland, OH 44106, USA. Electronic address: carisatilton@gmail.com.
authors.forename: Carisa A
authors.initials: CA
authors.lastname: Tilton
authors.forename: Caroline O
authors.initials: CO
authors.lastname: Tabler
authors.forename: Mark B
authors.initials: MB
authors.lastname: Lucera
authors.forename: Samantha L
authors.initials: SL
authors.lastname: Marek
authors.forename: Aiman A
authors.initials: AA
authors.lastname: Haqqani
authors.forename: John C
authors.initials: JC
authors.lastname: Tilton
mesh_terms: D015496:CD4-Positive T-Lymphocytes / Q000821:virology*; D002478:Cells, Cultured; D015870:Gene Expression; D017930:Genes, Reporter; D006678:HIV / Q000502:physiology*; D006801:Humans; D048348:Reverse Transcription; D013194:Staining and Labeling / Q000379:methods; D014773:Virology / Q000379:methods*; D016662:Virus Integration; D057646:Virus Uncoating
keywords: Fusion; HIV; LTR; Outcome; Post-entry event efficiency; Reporter virus
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