Deep molecular characterization of HIV-1 dynamics under suppressive HAART

• Published in: PLoS pathogens 2011-10, Vol.7 (10), p.e1002314
• Main Authors: Buzón, Maria J, Codoñer, Francisco M, Frost, Simon D W, Pou, Christian, Puertas, Maria C, Massanella, Marta, Dalmau, Judith, Llibre, Josep M, Stevenson, Mario, Blanco, Julià, Clotet, Bonaventura, Paredes, Roger, Martinez-Picado, Javier
• Format: Article
• Language: English
• Subjects: Anti-HIV Agents - pharmacology; Antiretroviral Therapy, Highly Active; Biology; Chemical properties; Disease Reservoirs; DNA; DNA, Circular - analysis; DNA, Circular - drug effects; DNA, Circular - genetics; DNA, Viral - analysis; DNA, Viral - drug effects; Drug therapy; Genetic aspects; Genetic testing; Haplotypes; Health aspects; Highly active antiretroviral therapy; HIV; HIV (Viruses); HIV Infections - blood; HIV Infections - drug therapy; HIV Infections - virology; HIV-1 - drug effects; HIV-1 - genetics; Host-Pathogen Interactions; Human immunodeficiency virus; Humans; Infections; Leukocytes, Mononuclear - drug effects; Leukocytes, Mononuclear - virology; Phylogenetics; Phylogeny; Physiological aspects; Plasma; Plasmids - drug effects; Plasmids - genetics; Population; Pyrrolidinones - pharmacology; Raltegravir Potassium; Sequence Analysis, DNA; Statistical methods; Viral Load
• ISSN: 1553-7374; 1553-7366; 1553-7374
• DOI: 10.1371/journal.ppat.1002314

In order to design strategies for eradication of HIV-1 from infected individuals, detailed insight into the HIV-1 reservoirs that persist in patients on suppressive antiretroviral therapy (ART) is required. In this regard, most studies have focused on integrated (proviral) HIV-1 DNA forms in cells circulating in blood. However, the majority of proviral DNA is replication-defective and archival, and as such, has limited ability to reveal the dynamics of the viral population that persists in patients on suppressive ART. In contrast, extrachromosomal (episomal) viral DNA is labile and as a consequence is a better surrogate for recent infection events and is able to inform on the extent to which residual replication contributes to viral reservoir maintenance. To gain insight into the diversity and compartmentalization of HIV-1 under suppressive ART, we extensively analyzed longitudinal peripheral blood mononuclear cells (PBMC) samples by deep sequencing of episomal and integrated HIV-1 DNA from patients undergoing raltegravir intensification. Reverse-transcriptase genes selectively amplified from episomal and proviral HIV-1 DNA were analyzed by deep sequencing 0, 2, 4, 12, 24 and 48 weeks after raltegravir intensification. We used maximum likelihood phylogenies and statistical tests (AMOVA and Slatkin-Maddison (SM)) in order to determine molecular compartmentalization. We observed low molecular variance (mean variability ≤0.042). Although phylogenies showed that both DNA forms were intermingled within the phylogenetic tree, we found a statistically significant compartmentalization between episomal and proviral DNA samples (P