A high throughput Cre-lox activated viral membrane fusion assay identifies pharmacological inhibitors of HIV entry.

Abstract:

:Enveloped virus entry occurs when viral and cellular membranes fuse releasing particle contents into the target cell. Human immunodeficiency virus (HIV) entry occurs by cell-free virus or virus transferred between infected and uninfected cells through structures called virological synapses. We developed a high-throughput cell-based assay to identify small molecule inhibitors of cell-free or virological synapse-mediated entry. An HIV clone carrying Cre recombinase as a Gag-internal gene fusion releases active Cre into cells upon viral entry activating a recombinatorial gene switch changing dsRed to GFP-expression. A screen of a 1998 known-biological profile small molecule library identified pharmacological HIV entry inhibitors that block both cell-free and cell-to-cell infection. Many top hits were noted as HIV inhibitors in prior studies, but not previously recognized as entry antagonists. Modest therapeutic indices for simvastatin and nigericin were observed in confirmatory HIV infection assays. This robust assay is adaptable to study HIV and heterologous viral pseudotypes.

journal_name

Virology

journal_title

Virology

authors

Esposito AM,Cheung P,Swartz TH,Li H,Tsibane T,Durham ND,Basler CF,Felsenfeld DP,Chen BK

doi

10.1016/j.virol.2015.10.013

subject

Has Abstract

pub_date

2016-03-01 00:00:00

pages

6-16

eissn

0042-6822

issn

1096-0341

pii

S0042-6822(15)00441-9

journal_volume

490

pub_type

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