AEGiS-14IAC: HIV drug resistance genotyping with DNA from peripheral blood mononuclear cells: involvement of latently infected methylated proviral DNA.

14th International AIDS Conference


Barcelona, Spain - July 7-12, 2002

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HIV drug resistance genotyping with DNA from peripheral blood mononuclear cells: involvement of latently infected methylated proviral DNA.

Int Conf AIDS 2002 Jul 7-12; 14:(abstract no. A10047)

Kudo S, Yoshizumi S, Miyoshi M, Iki S, Sawada H, Yano S, Takahashi Y
Hokkaido Institute of Public Health, Sapporo, Japan

BACKGROUND: The drug resistance mutations of HIV DNA in peripheral blood mononuclear cells (PBMC) are sometimes distinct from those of plasma HIV. To explain this phenomenon, we investigated the genotype of HIV isolated by co-culturing patient PBMC with those of healthy donors. Furthermore, we analyzed the methylation status of viral DNA in infected cells of PBMC.

METHODS: Sequence-based analysis of drug resistant mutations in reverse transcriptase and protease was conducted using plasma HIV, PBMC DNA, and HIV isolated from PBMC treated with or without DNA methyltransferase inhibitor 5-azacytidine (5-azaC). Methylation status of the LTR was analyzed by PCR using a methylation sensitive restriction enzyme HpaII.

RESULTS: Distinct drug resistance genotypes of HIV were observed in plasma and PBMC from a patient given multidrug therapy. The genotype of HIV isolated from PBMC by a standard method using IL2 was similar to that of plasma HIV. On the other hand, HIV isolates derived from the 5-azaC-treated PBMC showed the genotype similar to that of PBMC HIV-DNA. HpaII-PCR analysis of PBMC DNA revealed a significant methylated state of the HpaII sites of HIV-LTR DNA. Therefore, the methylated provirus represents a large fraction of the total proviral population in a viral reservoir of PBMC.

CONCLUSIONS: The discrepancy of drug resistance genotypes of HIV in plasma and PBMC might be explained by a higher proportion of HIV-infected cells harboring methylated provirus compared to that of HIV-infected cells harboring unmethylated provirus.

Keywords: AEGIS, HIV, HIV Infections, Proviruses, Anti-HIV Agents, DNA, Viral, Genotype, HIV Protease, HIV Seropositivity, Drug Resistance, RNA-Directed DNA Polymerase, HIV Protease Inhibitors, Greece, Human, geneticsKWDaegis,hiv,hivinfections,proviruses,anti-hivagents,dna,viral,genotype,hivprotease,hivseropositivity,drugresistance,rna-directeddnapolymerase,hivproteaseinhibitors,greece,human,genetics

020707
A10047

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