Department of Molecular Microbiology and Immunology, Keck School of Medicine of the University of Southern California, Los Angeles, California, USA.
Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, Minnesota, USA.
J Virol. 2019 May 1;93(10). doi: 10.1128/JVI.02086-18. Print 2019 May 15.
Combination anti-retroviral drug therapy (ART) potently suppresses HIV-1 replication but does not result in virus eradication or a cure. A major contributing factor is the long-term persistence of a reservoir of latently infected cells. To study this reservoir, we established a humanized mouse model of HIV-1 infection and ART suppression based on an oral ART regimen. Similar to humans, HIV-1 levels in the blood of ART-treated animals were frequently suppressed below the limits of detection. However, the limited timeframe of the mouse model and the small volume of available samples makes it a challenging model with which to achieve full viral suppression and to investigate the latent reservoir. We therefore used an latency reactivation assay that allows a semiquantitative measure of the latent reservoir that establishes in individual animals, regardless of whether they are treated with ART. Using this assay, we found that latently infected human CD4 T cells can be readily detected in mouse lymphoid tissues and that latent HIV-1 was enriched in populations expressing markers of T cell exhaustion, PD-1 and TIGIT. In addition, we were able to use the latency reactivation assay to demonstrate that HIV-specific TALENs can reduce the fraction of reactivatable virus in the latently infected cell population that establishes , supporting the use of targeted nuclease-based approaches for an HIV-1 cure. HIV-1 can establish latent infections that are not cleared by current antiretroviral drugs or the body's immune responses and therefore represent a major barrier to curing HIV-infected individuals. However, the lack of expression of viral antigens on latently infected cells makes them difficult to identify or study. Here, we describe a humanized mouse model that can be used to detect latent but reactivatable HIV-1 in both untreated mice and those on ART and therefore provides a simple system with which to study the latent HIV-1 reservoir and the impact of interventions aimed at reducing it.
联合抗逆转录病毒药物治疗（ART）能够强力抑制 HIV-1 的复制，但不能实现病毒清除或治愈。一个主要的促成因素是潜伏感染细胞的长期持续存在。为了研究这个储存库，我们建立了一个基于口服 ART 方案的 HIV-1 感染和 ART 抑制的人源化小鼠模型。与人类相似，ART 治疗动物的血液中的 HIV-1 水平经常被抑制到检测限以下。然而，由于小鼠模型的时间限制和可用样本量小，它是一个具有挑战性的模型，难以实现完全病毒抑制和研究潜伏储存库。因此，我们使用了一种潜伏再激活测定法，该方法可以半定量地测量个体动物中建立的潜伏储存库，而不管它们是否接受 ART 治疗。使用该测定法，我们发现潜伏感染的人 CD4 T 细胞可以在小鼠淋巴组织中轻易地检测到，并且潜伏 HIV-1 在表达 T 细胞衰竭标志物 PD-1 和 TIGIT 的群体中富集。此外，我们能够使用潜伏再激活测定法证明 HIV 特异性 TALENs 可以减少潜伏感染细胞群体中可再激活病毒的比例，这支持了使用靶向核酸酶方法实现 HIV-1 治愈。HIV-1 可以建立潜伏感染，这些感染不能被当前的抗逆转录病毒药物或机体的免疫反应清除，因此代表了治愈 HIV 感染个体的主要障碍。然而，潜伏感染细胞中病毒抗原的缺乏使其难以识别或研究。在这里，我们描述了一种人源化小鼠模型，该模型可用于检测未经治疗的小鼠和接受 ART 的小鼠中的潜伏但可再激活的 HIV-1，因此提供了一个简单的系统，可用于研究潜伏的 HIV-1 储存库以及旨在减少其的干预措施的影响。
