Kinetics of Human Immunodeficiency Virus Type 1 (HIV) DNA Integration in Acutely Infected Cells as Determined Using a Novel Assay for Detection of Integrated HIV DNA
- 15 November 2001
- journal article
- Published by American Society for Microbiology in Journal of Virology
- Vol. 75 (22), 11253-11260
- https://doi.org/10.1128/jvi.75.22.11253-11260.2001
Abstract
We have developed a novel linker-primer PCR assay for the detection and quantification of integrated human immunodeficiency virus type 1 (HIV) DNA. This assay reproducibly allowed the detection of 10 copies of integrated HIV DNA, in a background of 2 × 105 cell equivalents of human chromosomal DNA, without amplifying extrachromosomal HIV DNA. We have used this assay and a near-synchronous one-step T-cell infection model to investigate the kinetics of viral DNA accumulation following HIV infection. We report here that integrated HIV DNA started accumulating 1 h after the first appearance of extrachromosomal viral DNA and accounted for ∼10% of the total HIV DNA synthesized in the first round of viral replication. These results highlight the efficient nature of integrase-mediated HIV integration in infected T cells.Keywords
This publication has 55 references indexed in Scilit:
- Specific Inhibition of Human Immunodeficiency Virus Type 1 (HIV-1) Integration in Cell Culture: Putative Inhibitors of HIV-1 IntegraseAntimicrobial Agents and Chemotherapy, 2001
- Quantification of latent tissue reservoirs and total body viral load in HIV-1 infectionNature, 1997
- HIV-1 cDNA Integration: Requirement of HMG I(Y) Protein for Function of Preintegration Complexes In VitroCell, 1997
- Stepwise analysis of reverse transcription in a cell-to-cell human immunodeficiency virus infection model: kinetics and implicationsJournal of General Virology, 1995
- Kinetic Analysis of HIV-1 Early Replicative Steps in a Coculture SystemAIDS Research and Human Retroviruses, 1994
- Characterization of HIV Replication Complexes Early after Cell-to-Cell InfectionAIDS Research and Human Retroviruses, 1993
- Synthesis of Human Immunodeficiency Virus DNA In a Cell-to-Cell Transmission ModelAIDS Research and Human Retroviruses, 1992
- Productive human immunodeficiency virus type 1 (HIV-1) infection of nonproliferating human monocytes.The Journal of Experimental Medicine, 1991
- Biological and biochemical characterization of a cloned Leu-3- cell surviving infection with the acquired immune deficiency syndrome retrovirus.The Journal of Experimental Medicine, 1986
- Selective extraction of polyoma DNA from infected mouse cell culturesJournal of Molecular Biology, 1967