Brain Macrophages in Simian Immunodeficiency Virus-Infected, Antiretroviral-Suppressed Macaques: a Functional Latent Reservoir.
Brain Macrophages in Simian Immunodeficiency Virus-Infected, Antiretroviral-Suppressed Macaques: a Functional Latent Reservoir.
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DOI：
10.1128/mbio.01186-17
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发表时间：
2017-08-15
期刊：
影响因子：
6.4
通讯作者：
Gama L
中科院分区：
文献类型：
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作者：
Avalos CR;Abreu CM;Queen SE;Li M;Price S;Shirk EN;Engle EL;Forsyth E;Bullock BT;Mac Gabhann F;Wietgrefe SW;Haase AT;Zink MC;Mankowski JL;Clements JE;Gama L
人类免疫缺陷病毒(HIV)感染的治疗需要了解携带病毒的细胞和解剖部位，这些部位有助于在治疗中断时导致病毒反弹。尽管接受了抗逆转录病毒疗法(ART)，但在接受ART治疗的艾滋病毒感染者中，报告了艾滋病毒相关的神经认知障碍(HAND)。HIV感染者脑脊液中巨噬细胞激活和神经元损伤的生物标志物表明HIV在大脑中的持续影响，并提示中枢神经系统(CNS)可能是病毒库。使用HIV脑炎和艾滋病的猴免疫缺陷病毒(SIV)/猕猴模型，我们评估了感染细胞是否在脑内持续存在，尽管抗逆转录病毒治疗。用ART对8只感染SIV的猪尾猕猴进行病毒抑制，并纵向分析血浆和脑脊液病毒血症水平。为了评估病毒在这些猕猴脑巨噬细胞(BRMΦ)中是否持续存在，我们使用巨噬细胞定量病毒生长试验(MΦ-QVOA)、聚合酶链式反应和原位杂交(ISH)来测量感染细胞的频率以及脑中病毒的RNA和DNA水平。虽然在所有动物中都检测到了病毒DNA，但在受抑制的猕猴脑组织中检测不到病毒RNA。MΦ-QVOA显示，大多数被抑制的动物含有潜伏感染的BRMΦ。我们还表明，在MΦ-QVOAs中产生的病毒具有复制能力，这表明潜伏感染的BRMΦ能够在治疗中断后重建生产性感染。这份报告首次证实了在ART抑制的猕猴的BRMΦ中存在具有复制能力的SIV，并表明在接受ART治疗的SIV感染的猕猴中，巨噬细胞中病毒潜伏的高度争论的问题，至少在大脑中，已经得到了解决。静息的CD4+T细胞是目前唯一符合潜伏库定义的细胞。然而，最近的证据表明，尽管抗逆转录病毒治疗，艾滋病毒/SIV感染的巨噬细胞仍然存在。在HIV感染者和抑制ART方案的SIV感染猕猴的脑脊液中观察到巨噬细胞激活和神经元损伤的标志，表明中枢神经系统存在持续的病毒感染和潜伏感染。关于脑巨噬细胞是否代表复制能力强的病毒的潜在来源，能够在治疗中断时重新建立感染，存在争议。在这项研究中，我们证明了SIV感染的经ART治疗的猕猴大脑中存在潜在的巨噬细胞储存库，并使用我们建立的生长试验对该储存库进行了分析，以定量含有具有复制能力的SIV基因组的巨噬细胞。我们的结果支持除了静止的CD4+T细胞外，还存在其他潜伏库的想法，并强调了巨噬细胞在制定根除艾滋病毒的策略中的重要性。
A human immunodeficiency virus (HIV) infection cure requires an understanding of the cellular and anatomical sites harboring virus that contribute to viral rebound upon treatment interruption. Despite antiretroviral therapy (ART), HIV-associated neurocognitive disorders (HAND) are reported in HIV-infected individuals on ART. Biomarkers for macrophage activation and neuronal damage in cerebrospinal fluid (CSF) of HIV-infected individuals demonstrate continued effects of HIV in brain and suggest that the central nervous system (CNS) may serve as a viral reservoir. Using a simian immunodeficiency virus (SIV)/macaque model for HIV encephalitis and AIDS, we evaluated whether infected cells persist in brain despite ART. Eight SIV-infected pig-tailed macaques were virally suppressed with ART, and plasma and CSF viremia levels were analyzed longitudinally. To assess whether virus persisted in brain macrophages (BrMΦ) in these macaques, we used a macrophage quantitative viral outgrowth assay (MΦ-QVOA), PCR, and in situ hybridization (ISH) to measure the frequency of infected cells and the levels of viral RNA and DNA in brain. Viral RNA in brain tissue of suppressed macaques was undetectable, although viral DNA was detected in all animals. The MΦ-QVOA demonstrated that the majority of suppressed animals contained latently infected BrMΦ. We also showed that virus produced in the MΦ-QVOAs was replication competent, suggesting that latently infected BrMΦ are capable of reestablishing productive infection upon treatment interruption. This report provides the first confirmation of the presence of replication-competent SIV in BrMΦ of ART-suppressed macaques and suggests that the highly debated issue of viral latency in macrophages, at least in brain, has been addressed in SIV-infected macaques treated with ART. Resting CD4+ T cells are currently the only cells that fit the definition of a latent reservoir. However, recent evidence suggests that HIV/SIV-infected macrophages persist despite ART. Markers of macrophage activation and neuronal damage are observed in the CSF of HIV-infected individuals and of SIV-infected macaques on suppressive ART regimens, suggesting that the CNS has continued virus infection and latent infection. A controversy exists as to whether brain macrophages represent a latent source of replication-competent virus capable of reestablishing infection upon treatment interruption. In this study, we demonstrated the presence of the latent macrophage reservoir in brains of SIV-infected ART-treated macaques and analyzed the reservoir using our established outgrowth assay to quantitate macrophages harboring replication-competent SIV genomes. Our results support the idea of the existence of other latent reservoirs in addition to resting CD4+ T cells and underscore the importance of macrophages in developing strategies to eradicate HIV.