High-throughput screening using pseudotyped lentiviral particles: A strategy for the identification of HIV-1 inhibitors in a cell-based assay

• Published in: Antiviral research Vol. 81; no. 3; pp. 239 - 247
• Main Authors: Garcia, Jean-Michel, Gao, Anhui, He, Pei-Lan, Choi, Joyce, Tang, Wei, Bruzzone, Roberto, Schwartz, Olivier, Naya, Hugo, Nan, Fa-Jun, Li, Jia, Altmeyer, Ralf, Zuo, Jian-Ping
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 01.03.2009; Elsevier; Elsevier Masson
• Subjects: Anti-HIV Agents; Anti-HIV Agents - pharmacology; Antibiotics. Antiinfectious agents. Antiparasitic agents; Antiviral agents; Biological and medical sciences; Cell Line; Cocktail library; Drug Evaluation, Preclinical; Drug Evaluation, Preclinical - methods; High-throughput screening; HIV; HIV-1; HIV-1 - drug effects; HIV-1 - genetics; HTS; Human immunodeficiency virus; Human immunodeficiency virus 1; Human viral diseases; Humans; Immunodeficiencies; Immunodeficiencies. Immunoglobulinopathies; Immunopathology; Infectious diseases; Life Sciences; Medical sciences; Microbiology and Parasitology; NNRTI; Pharmacology. Drug treatments; Pseudotyped lentiviral particles; Viral diseases; Viral diseases of the lymphoid tissue and the blood. Aids; Virus Replication; Virus Replication - drug effects; 3TC; SFV; d4T; DMSO; HTS; ddI; NNRTI; High-throughput screening; NVP; MTT; PDB; GFP; VSV-G; HIV; Cocktail library; FCS; CPE; Pseudotyped lentiviral particles; BSL; MDR; AZT; Luc; CPRG; TCM; High throughput screening; Antiretroviral agent; HIV-1 virus; Non nucleoside compound; Identification; Reverse transcriptase inhibitor; Antiviral; Cell; Immunopathology; Statistical analysis; Retroviridae; AIDS; Immune deficiency; Lentivirus; In vitro; Infection; Virus; Viral disease; Strategy; Human immunodeficiency virus
• ISSN: 0166-3542; 1872-9096
• DOI: 10.1016/j.antiviral.2008.12.004

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.