AEGiS-15IAC: Analysis of HIV-1 defective genomes in a cohort of HIV-1 infected intravenous drug abusers (IVDA) from European Russia.

15th International AIDS Conference


Bangkok, Thailand - July 11-16, 2004

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Analysis of HIV-1 defective genomes in a cohort of HIV-1 infected intravenous drug abusers (IVDA) from European Russia.

Int Conf AIDS 2004 Jul 11-16; 15:(abstract no. ThOrA1366)

Papuashvili MN, Morozov VA, Shcherbakova TI
Advanced Medical Researches, Moscow, Russian Federation

BACKGROUND: Defective genomes (DG) are frequent in HIV-1 infected individuals with active virus replication and a high virus load. DG with large internal deletions might not be detected using HIV-1 diagnostic primers. Putative role of DG accumulating during disease progression was not investigated in details previously. In vivo follow up of DG during a significant period of time might be of interest and could shed light on virus variability and replication.

METHODS: DNA extracted from blood of 99 HIV-1 positive therapy-naïve IVDA infected 1-3 years ago. Cell cultures, PCR, nested PCR, cloning and sequencing.

RESULTS: Before screening for DG in DNA specimens, HIV-1 nearly full size genome presence was confirmed by PCR with a set of diagnostic (gag, pol, env) primers. Phylogenetic analysis (neighbor joining method) indicated that most proviruses clusters with subtype A. By nested PCR (with primers for DG detection and corresponding to the 5'and 3' fragments of the provirus), HIV-1 DG were revealed in 30/99 individuals. DG genome looks as follows: 5'LTR-5' gag -3'env. The size of DG varied from 0, 27 kb to nearly 2 kb.

CONCLUSIONS: 30% of HIV-1 positive IVDA contained DG. In vitro co-cultivation of the PBMCs from all "DG positive" individuals with human mononuclear cells, results in gradual decrease of DG. This phenomenon is likely attributed to the negative influence of DG on cell survival in vitro. In vitro co-cultivation of HIV-1 DG negative cells from HIV-1 infected individuals is under way. Regular monitoring for DG transcripts and putative DG modifications would be performed in the future to extend our knowleDGe on DG impact in pathogenesis of HIV-1 infection.

Keywords: AEGIS, HIV-1, Substance Abuse, Intravenous, Genes, gag, HIV Infections, Genes, env, Genes, pol, Proviruses, Polymerase Chain Reaction, Genome, Genome, Viral, Russia, Humans, In Vitro, genetics

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ThOrA1366

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