HIV-1 Infection and First Line ART Induced Differential Responses in Mitochondria from Blood Lymphocytes and Monocytes: The ANRS EP45 ‘‘Aging’’ Study Sophie Perrin1,2, Jonathan Cremer1,2, Patrice Roll1,2, Olivia Faucher3, Ame´ lie Me´ nard3, Jacques Reynes4, Pierre Dellamonica5, Alissa Naqvi5, Joe¨ lle Micallef6, Elisabeth Jouve6, Catherine Tamalet7,8, Caroline Solas9,10, Christel Pissier9,10, Isabelle Arnoux11, Corine Nicolino-Brunet12, Le´ on Espinosa8¤, Nicolas Le´ vy1,13, Elise Kaspi1,2, Andre´ e Robaglia-Schlupp1,2, Isabelle Poizot-Martin3, Pierre Cau1,2* 1 Inserm UMR 910, Aix-Marseille Univ, Marseille, France, 2 Laboratoire de Biologie Cellulaire, CHU (Centre Hospitalier Universitaire) La Timone AP-HM (Assistance Publique - H oˆpitaux de Marseille), Marseille, France, 3 Service d’Immuno-He´matologie Clinique, CHU (Centre Hospitalier Universitaire) Sainte Marguerite AP-HM (Assistance Publique - H oˆpitaux de Marseille), Marseille, France, 4 De´partement des Maladies Infectieuses et Tropicales, CHRU (Centre Hospitalier Re´gional et Universitaire) Gui-de-Chauliac, Montpellier, France, 5 Service d’Infectiologie, CHU (Centre Hospitalier Universitaire) L’Archet 1, Nice, France, 6 Centre d’Investigation Clinique - Unite´ de Pharmacologie Clinique et d’Evaluations The´rapeutiques (CIC-UPCET), CHU (Centre Hospitalier Universitaire) La Timone AP-HM (Assistance Publique - Hoˆ pitaux de Marseille), Marseille, France, 7 Fe´de´ration de Microbiologie Clinique, CHU (Centre Hospitalier Universitaire) La Timone AP-HM (Assistance Publique - Hoˆ pitaux de Marseille), Marseille, France, 8 URMITE CNRS-IRD UMR 6236, Aix-Marseille Univ, Marseille, France, 9 Laboratoire de Pharmacocine´tique et de Toxicologie, CHU (Centre Hospitalier Universitaire) La Timone AP-HM (Assistance Publique - H oˆpitaux de Marseille), Marseille, France, 10 Inserm UMR U911, Aix-Marseille Univ, Marseille, France, 11 Laboratoire d’He´matologie, CHU (Centre Hospitalier Universitaire) La Timone AP-HM (Assistance Publique - Hoˆ pitaux de Marseille), Marseille, France, 12 Laboratoire d’He´matologie, CHU (Centre Hospitalier Universitaire) La Conception AP-HM (Assistance Publique - H oˆpitaux de Marseille), Marseille, France, 13 Laboratoire de Ge´netique Mole´culaire, CHU (Centre Hospitalier Universitaire) La Timone AP-HM (Assistance Publique - Hoˆ pitaux de Marseille), Marseille, France Abstract Background: The ANRS EP45 ‘‘Aging’’ study investigates the cellular mechanisms involved in the accelerated aging of HIV-1 infected and treated patients. The data reported focus on mitochondria, organelles known to be involved in cell senescence. Methods: 49 HIV-1 infected patients untreated with antiretroviral therapy, together with 49 seronegative age- and sex- matched control subjects and 81 HIV-1 infected and treated patients, were recruited by 3 AIDS centres (Marseille, Montpellier, Nice; France; http://clinicaltrials.gov/, NCT01038999). In more than 88% of treated patients, the viral load was 3 ,40 copies/ml and the CD4+ cell count was .500/mm . ROS (reactive oxygen species) production and DYm (inner membrane potential) were measured by flow cytometry in blood lymphocytes and monocytes (functional parameters). Three mitochondrial network quantitative morphological parameters were computed using confocal microscopy and image analysis. Three PBMC mitochondrial proteins (porin and subunits 2 and 4 of cytochrome C oxidase encoded by mtDNA or nuclear DNA, respectively) were analysed by western blotting. Results: Quantitative changes in PBMC mitochondrial proteins were not induced by either HIV-1 infection or ART. Discriminant analysis integrating functional (ROS production and DYm) or morphological (network volume density, fragmentation and branching) parameters revealed HIV-1 infection and ART differential effects according to cell type. First line ART tended to rescue lymphocyte mitochondrial parameters altered by viral infection, but induced slight changes in monocytes. No statistical difference was found between the effects of three ART regimens on mitochondrial parameters. Correlations between functional parameters and viral load confirmed the damaging effects of HIV-1 in lymphocyte mitochondria. Conclusions: In patients considered to be clinically stable, mitochondria exhibited functional and morphological modifications in PBMCs resulting from either direct or indirect effects of HIV-1 infection (lymphocytes), or from first line ART (monocytes). Together with other tissue impairments, these changes may contribute to global aging. Trial Registration: ClinicalTrials.gov NCT01038999 Citation: Perrin S, Cremer J, Roll P, Faucher O, Me´nard A, et al. (2012) HIV-1 Infection and First Line ART Induced Differential Responses in Mitochondria from Blood Lymphocytes and Monocytes: The ANRS EP45 ‘‘Aging’’ Study. PLoS ONE 7(7): e41129. doi:10.1371/journal.pone.0041129 Editor: Patricia Fitzgerald-Bocarsly, University of Medicine and Dentistry of New Jersey - New Jersey Medical School, United States of America Received March 23, 2012; Accepted June 18, 2012; Published July 19, 2012 Copyright: 2012 Perrin et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was supported by the French National Agency for Research on AIDS and Viral Hepatitis (ANRS, http://www.anrs.fr/) [ANRS EP45 ‘‘Aging’’ to PC]. SP was a recipient of a 3-year Ph.D. grant from ANRS [ANRS EP45 ‘‘Aging’’ to PC] and a fourth additional Ph.D. year from Sidaction (http://www.sidaction.org/) grant [B/22-2-02032 to SP]. SP received a 3-month transition grant from ADEREM (Biological and medical research supportive association for Marseille hospital centres, http://aderem.fr/index.htm). JC was a recipient of a grant from Sidaction [A/19-3-01487 to PC]. PC was a recipient of grants from GlaxoSmithKline and from Boehringen Mannheim. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: GlaxoSmithKline and Boehringen Mannheim grants were received. This does not alter the authors’ adherence to all PLoS ONE policies on sharing data and materials. The funding was not related to employment, consultancy, patents and product in development or marketed products. * E-mail: pierre.cau@univ-amu.fr ¤ Current address: Plateforme de Biophotonique applique´e a` la Microbiologie, LCB CNRS UPR9043, Marseille, France PLoS ONE | www.plosone.org 1 July 2012 | Volume 7 | Issue 7 | e41129