title: Evolution of 2-long terminal repeat (2-LTR) episomal HIV-1 DNA in raltegravir-treated patients and in in vitro infected cells.
abstract: OBJECTIVE
  Our aim was to analyse the evolution of HIV-1 2-long terminal repeat (2-LTR) circular DNA in vitro and ex vivo in the presence of raltegravir.
  
  
  METHODS
  Twenty-five patients starting a raltegravir-based regimen were included. Total HIV-1 DNA and 2-LTR DNA were quantified at baseline and in follow-up samples up to month 12. The effect of raltegravir on the formation of 2-LTR circles was evaluated in HeLa P4 cells. The effect of raltegravir was also investigated by sequence analysis of the 2-LTR circle junctions.
  
  
  RESULTS
  Among 21 patients with undetectable 2-LTR DNA at baseline, 7 had detectable 2-LTR DNA during the follow-up. Three of four patients with detectable 2-LTR DNA at baseline had undetectable 2-LTR DNA during the follow-up (P = 0.27). The mean 2-LTR level increased significantly (+0.07 log(10)/month, P = 0.02) in raltegravir-treated patients, and a 2-LTR increase was also observed in raltegravir-treated HeLa P4 cells, with a peak at 3 days post-infection. 2-LTR DNA showed a high prevalence of deletions ex vivo (64.5%) and in vitro (50%) in the presence of raltegravir, which was not statistically different from the prevalence in untreated patients or cells.
  
  
  CONCLUSIONS
  In antiretroviral-experienced patients receiving raltegravir, 2-LTR DNA increased while total HIV-1 DNA decreased over time. The frequent rearrangements found in 2-LTR sequences warrant further investigations to determine the dynamics of evolution of unintegrated HIV-1 DNA.
authors.affiliation: Laboratory of Virology, EA2968, University of Bordeaux 2, Bordeaux, France. sandrine.reigadas@chu-bordeaux.fr
authors.forename: Sandrine
authors.initials: S
authors.lastname: Reigadas
authors.forename: Marie-Line
authors.initials: ML
authors.lastname: Andréola
authors.forename: Linda
authors.initials: L
authors.lastname: Wittkop
authors.forename: Ophélie
authors.initials: O
authors.lastname: Cosnefroy
authors.forename: Guerric
authors.initials: G
authors.lastname: Anies
authors.forename: Patricia
authors.initials: P
authors.lastname: Recordon-Pinson
authors.forename: Rodolphe
authors.initials: R
authors.lastname: Thiébaut
authors.forename: Bernard
authors.initials: B
authors.lastname: Masquelier
authors.forename: Hervé
authors.initials: H
authors.lastname: Fleury
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology / Q000627:therapeutic use*; D004270:DNA, Circular / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D019143:Evolution, Molecular; D015658:HIV Infections / Q000188:drug therapy* / Q000821:virology*; D016325:HIV Long Terminal Repeat / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D010957:Plasmids; D011760:Pyrrolidinones / Q000494:pharmacology / Q000627:therapeutic use*; D000068898:Raltegravir Potassium; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA; D019562:Viral Load
chemical_list: D019380:Anti-HIV Agents; D004270:DNA, Circular; D004279:DNA, Viral; D011760:Pyrrolidinones; D000068898:Raltegravir Potassium