High-throughput screening using pseudotyped lentiviral particles: a strategy for the identification of HIV-1 inhibitors in a cell-based assay.
Two decades after its discovery the human immunodeficiency virus (HIV) is still spreading worldwide and killing millions. There are 25 drugs formally approved for HIV currently on the market, but side effects as well as the emergence of HIV strains showing single or multiple resistances to current drug-therapy are causes for concern. Furthermore, these drugs target only 4 steps of the viral cycle, hence the urgent need for new drugs and also new targets. In order to tackle this problem, we have devised a cell-based assay using lentiviral particles to look for post-entry inhibitors of HIV-1. We report here the assay development, validation as well as confirmation of the hits using both wild-type and drug-resistant HIV-1 viruses. The screening was performed on an original library, rich in natural compounds and pure molecules from Traditional Chinese Medicine pharmacopoeia, which had never been screened for anti-HIV activity. The identified hits belong to four chemical sub-families that appear to be all non-nucleoside reverse transcriptase inhibitors (NNRTIs). Secondary tests with live viruses showed that there was good agreement with pseudotyped particles, confirming the validity of this approach for high-throughput drug screens. This assay will be a useful tool that can be easily adapted to screen for inhibitors of viral entry.
Garcia
Jean-Michel
JM
HKU-Pasteur Research Centre Ltd., Hong Kong, China. jmgarcia@hkucc.hku.hk
Gao
Anhui
A
He
Pei-Lan
PL
Choi
Joyce
J
Tang
Wei
W
Bruzzone
Roberto
R
Schwartz
Olivier
O
Naya
Hugo
H
Nan
Fa-Jun
FJ
Li
Jia
J
Altmeyer
Ralf
R
Zuo
Jian-Ping
JP
D019380:Anti-HIV Agents / Q000494:pharmacology*; D002460:Cell Line; D004353:Drug Evaluation, Preclinical / Q000379:methods; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D014779:Virus Replication / Q000187:drug effects*
D019380:Anti-HIV Agents