Specific inhibition of human immunodeficiency virus type 1 (HIV-1) integration in cell culture: putative inhibitors of HIV-1 integrase.
To study the effect of potential human immunodeficiency virus type 1 (HIV-1) integrase inhibitors during virus replication in cell culture, we used a modified nested Alu-PCR assay to quantify integrated HIV DNA in combination with the quantitative analysis of extrachromosomal HIV DNA. The two diketo acid integrase inhibitors (L-708,906 and L-731,988) blocked the accumulation of integrated HIV-1 DNA in T cells following infection but did not alter levels of newly synthesized extrachromosomal HIV DNA. In contrast, we demonstrated that L17 (a member of the bisaroyl hydrazine family of integrase inhibitors) and AR177 (an oligonucleotide inhibitor) blocked the HIV replication cycle at, or prior to, reverse transcription, although both drugs inhibited integrase activity in cell-free assays. Quercetin dihydrate (a flavone) was shown to not have any antiviral activity in our system despite reported anti-integration properties in cell-free assays. This refined Alu-PCR assay for HIV provirus is a useful tool for screening anti-integration compounds identified in biochemical assays for their ability to inhibit the accumulation of integrated HIV DNA in cell culture, and it may be useful for studying the effects of these inhibitors in clinical trials.
Vandegraaff
N
N
National Centre for HIV Virology Research, Infectious Diseases Laboratories, Institute of Medical and Veterinary Science, Adelaide, Australia 5000. nicholas.vandegraaf@imvs.sa.gov.au
Kumar
R
R
Hocking
H
H
Burke
T R
TR
Mills
J
J
Rhodes
D
D
Burrell
C J
CJ
Li
P
P
D000090:Acetoacetates / Q000494:pharmacology; D002478:Cells, Cultured; D004279:DNA, Viral / Q000187:drug effects* / Q000502:physiology; D019427:HIV Integrase / Q000187:drug effects / Q000378:metabolism*; D019428:HIV Integrase Inhibitors / Q000494:pharmacology*; D015497:HIV-1 / Q000187:drug effects* / Q000201:enzymology / Q000502:physiology; D006801:Humans; D006834:Hydrazines / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D009841:Oligonucleotides / Q000494:pharmacology; D014779:Virus Replication
D000090:Acetoacetates; D004279:DNA, Viral; D019428:HIV Integrase Inhibitors; D006834:Hydrazines; C403887:L 708906; C435766:L17 compound; D009841:Oligonucleotides; C098172:T 30177; D019427:HIV Integrase