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Antimicrobial Agents and Chemotherapy, September 2001, p. 2510-2516, Vol. 45, No. 9
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.9.2510-2516.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Specific Inhibition of Human Immunodeficiency Virus Type 1 (HIV-1) Integration in Cell Culture: Putative Inhibitors of HIV-1 Integrase

Nick Vandegraaff,1,2,* Raman Kumar,1 Helen Hocking,1 Terrence R. Burke Jr.,3 John Mills,4 David Rhodes,5 Christopher J. Burrell,1,2 and Peng Li1

National Centre for HIV Virology Research, Infectious Diseases Laboratories, Institute of Medical and Veterinary Science,1 and Department of Molecular Biosciences, University of Adelaide, North Terrace,2 Adelaide, Australia 5000; Laboratory of Medicinal Chemistry, Division of Basic Sciences, National Cancer Institute, Bethesda, Maryland 208923; National Centre for HIV Virology Research, Macfarlane Burnet Centre for Medical Research, Fairfield, Victoria, Australia 31414; and Amrad Operations, Richmond, Victoria, Australia, 31215

Received 20 February 2001/Returned for modification 21 May 2001/Accepted 11 June 2001

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.

* Corresponding author. Mailing address: National Centre for HIV Virology Research, Infectious Diseases Laboratories, Institute of Medical and Veterinary Science, Frome Road, Adelaide, Australia 5000. Phone: 61 8 82223574. Fax: 61 8 82223543. E-mail: nicholas.vandegraaf{at}imvs.sa.gov.au.

Antimicrobial Agents and Chemotherapy, September 2001, p. 2510-2516, Vol. 45, No. 9
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.9.2510-2516.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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