HIV-1 Infection and First Line ART Induced Differential Responses in Mitochondria from Blood Lymphocytes and Monocytes: The ANRS EP45 “Aging” Study

BackgroundThe 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. Methods49 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 500/mm3. ROS (reactive oxygen species) production and ΔΨm (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. ResultsQuantitative changes in PBMC mitochondrial proteins were not induced by either HIV-1 infection or ART. Discriminant analysis integrating functional (ROS production and ΔΨm) 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. ConclusionsIn 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 RegistrationClinicalTrials.gov NCT01038999 NCT01038999

研究背景：ANRS EP45“衰老”研究旨在探究HIV-1感染且接受治疗的患者加速衰老的细胞机制。本次报告的数据聚焦于线粒体——一种已知与细胞衰老相关的细胞器。
研究方法：本研究由法国马赛、蒙彼利埃、尼斯的3家艾滋病研究中心招募受试者：49名未接受抗逆转录病毒疗法（antiretroviral therapy，ART）的HIV-1感染者、49名血清学阴性且年龄、性别匹配的健康对照者，以及81名HIV-1感染且接受治疗的患者，试验注册于ClinicalTrials.gov（编号NCT01038999）。在88%以上的接受治疗的患者中，其病毒载量为500/mm³。采用流式细胞术检测血液淋巴细胞与单核细胞的活性氧（reactive oxygen species，ROS）生成量以及线粒体膜电位（inner membrane potential，ΔΨm），此为功能学指标。通过共聚焦显微镜与图像分析软件，计算得到3项线粒体网络定量形态学参数。采用蛋白质印迹法（western blotting）检测3项外周血单个核细胞（peripheral blood mononuclear cell，PBMC）线粒体蛋白：分别为孔蛋白（porin）以及由线粒体DNA（mitochondrial DNA，mtDNA）和核DNA编码的细胞色素C氧化酶亚基2与亚基4。
研究结果：HIV-1感染或ART治疗均未引起PBMC线粒体蛋白的定量变化。整合功能学指标（ROS生成量与ΔΨm）或形态学指标（线粒体网络体积密度、碎片化程度与分支数）的判别分析结果显示，HIV-1感染与ART治疗对不同细胞类型的影响存在差异。一线ART治疗可在一定程度上逆转病毒感染所致的淋巴细胞线粒体参数异常，但会对单核细胞造成轻微影响。三种ART治疗方案对线粒体参数的影响未发现统计学差异。功能学指标与病毒载量的相关性分析证实了HIV-1对淋巴细胞线粒体的损伤作用。
研究结论：在临床状态稳定的患者中，PBMC线粒体的功能与形态学改变，可由HIV-1感染（对淋巴细胞而言，为直接或间接作用）或一线ART治疗（对单核细胞而言）所导致。此类改变与其他组织损伤共同作用，可能参与全身性衰老的发生发展。
试验注册：本试验注册于ClinicalTrials.gov，编号为NCT01038999。