AEGiS-15IAC: Dynamics of linear unintegrated and integrated HIV-1 DNA in vitro and in vivo.

15th International AIDS Conference

Bangkok, Thailand - July 11-16, 2004

Dynamics of linear unintegrated and integrated HIV-1 DNA in vitro and in vivo.

Int Conf AIDS 2004 Jul 11-16; 15:(abstract no. ThOrA1400)

Koelsch K, Camposs-Soto P, Little S, Ignacio C, Havlir D, Young K, Shafer R, Robbins G, D'Aquila R, Haubrich R, Richman D, Spina C, Wong J
University of California, San Diego, La Jolla, United States

BACKGROUND: In HIV-1 infected patients HAART can lead to virus suppression (<50 RNA copies/ml plasma), but HIV DNA persists in various cell types. Recent studies report a potential link between HIV DNA levels and disease progression/treatment outcome. Integrated and circular HIV DNA are thought to be very stable, the stability of linear unintegrated forms has yet to be determined in vivo. Conventional HIV DNA assays cannot distinguish between these molecular forms.

METHODS: We analyzed HIV plasma RNA, CD4 cells and linear unintegrated and integrated HIV DNA in PBMC`s from acutely and chronically infected individuals (San Diego subjects, AIEDRP or ACTG 384 studies) with and without virus suppression. We also analyzed decay dynamics of total vs. integrated HIV DNA in resting primary CD4 T cells in vitro. HIV RNA and CD4 cell numbers were determined with standard commercial assays. Integrated and unintegrated HIV DNA was analyzed using an adaption of the Roche DNA Amplicor ó ä ó assay after gel separation of the DNA fractions based on molecular size, a procedure that is unaffected by location of integration sites.

RESULTS: HIV RNA and total HIV DNA decayed rapidly in individuals with virus suppression. Ratios of total to integrated HIV DNA copies were high during active replication (3:1) but diminished with virus suppression (=1 or less). In untreated patients the values for the different HIV DNA fractions remained relatively stable over 1-2 years. Linear unintegrated HIV DNA showed a rapid decay (t1/2 = 2.3-5.2 days) in vitro. Conclusion Linear unintegrated HIV DNA is labile in vitro and total HIV DNA decays rapidly with suppression of virus replication in vivo. Clearance of HIV DNA in the first six months of therapy likely reflects loss of unintegrated HIV DNA genomes. Values for total HIV-DNA in PBMC DNA after prolonged virus suppression largely represent integrated HIV genomes.

Keywords: AEGIS, HIV-1, HIV, Acquired Immunodeficiency Syndrome, HIV Infections, Anti-HIV Agents, CD4-Positive T-Lymphocytes, Antiretroviral Therapy, Highly Active, Virus Replication, HIV Long Terminal Repeat, HIV Core Protein p24, HIV Seropositivity, DNA, Circular, HIV Protease Inhibitors, In Vitro, Humans, virology, genetics, immunology

040711
ThOrA1400