Virus production from HIV reservoirs

描述（由申请人提供）：病毒库的无限期持续存在，有能力推动病毒反弹，是消除艾滋病毒感染的主要障碍。确定HIV-1的储藏库和HIV-1复制未被抗逆转录病毒（ARV）治疗（ART）完全抑制的庇护所将有助于探索改善治疗效果的新方法，并将对未来从体内消除HIV-1的努力至关重要。尽管进行了广泛的研究，但在接受抗逆转录病毒药物治疗的宿主体内保留艾滋病毒储存库和庇护所的位置和机制仍然知之甚少。从历史上看，了解病毒宿主的努力集中在容易接近的组织部位，如体循环、浅表淋巴结和生殖器液体。最近，一些小型研究试图检查其他部位，如大肠和小肠的肠道相关淋巴组织（GALT）、细支气管肺泡灌洗液、脑脊液和肾脏。然而，我们和其他人已经证明，一旦治疗停止或失败，外周血和淋巴结内的潜伏感染细胞并不是反弹病毒的重要来源，这些反弹病毒群的起源仍不清楚。因此，迫切需要对受感染宿主内的病毒库进行全面调查。为此，我们开发了创新的计算工具，用于从病毒基因序列数据中识别病毒库和区室。鉴定储存库和评估储存库保留机制的理想材料来源将是侵入性检查GALT（所有t淋巴细胞的bb0 - 65%是局部的，并且是HIV-1感染早期病毒复制的主要部位）和尸体解剖来源的组织，这些组织来自于在抑制性抗逆转录病毒治疗期间死亡的个体。目前，由于很难找到符合这些标准的受试者，这些研究领域实际上尚未被探索。然而，我们已经确定并获得了几个符合我们标准的人的组织，并获得了更多在没有接受抗逆转录病毒治疗的情况下死于艾滋病的对照组的尸体解剖。在此，我们建议在抗逆转录病毒治疗期间系统地探索整个身体的潜伏病毒储存库和活跃病毒复制的庇护所。我们建议在抑制性抗逆转录病毒治疗期间从GALT中获得HIV-1 RNA和DNA序列，并解剖在接受或不接受抑制性抗逆转录病毒治疗期间死亡的个体的组织，评估病毒库特征的序列，并识别具有正在进行的病毒复制庇护所特征的组织。

DESCRIPTION (provided by applicant): The indefinite persistence of virus reservoirs, with the capacity to fuel virus rebound, is the major obstacle to eliminating HIV infection. Identifying reservoirs of HIV-1 and sanctuaries where HIV-1 replication is not completely suppressed by antiretroviral (ARV) therapy (ART) will facilitate the exploration of new approaches to improve treatment outcomes, and will be critical to future efforts to eliminate HIV-1 from the body. Despite extensive study, the location and mechanisms for retention of HIV reservoirs and sanctuaries in the ARV treated host remain poorly understood. Historically, efforts to understand virus reservoirs have centered on easily accessible tissue sites such as the systemic circulation, superficial lymph nodes, and genital fluids. More recently, a few small studies have attempted to examine other sites such as Gut-associated lymphoid tissues (GALT) in the large and small bowel, bronchioalveolar lavage, CSF and kidney. However, we and others have demonstrated that latently infected cells within the peripheral blood and lymph nodes are not significant sources of rebounding virus once therapy is withdrawn or fails, and the origins of these rebounding virus populations remain unclear. A comprehensive investigation of virus reservoirs within the infected host is therefore urgently needed. To this end, we have developed innovative computational tools for the identification of virus reservoirs and compartments from viral gene sequence data. The ideal source of materials for identifying reservoirs, and for assessing mechanisms of retention of reservoirs, would be an invasive examination of both GALT (where >65% of all T-lymphocytes are localized, and the major site of virus replication early in HIV-1 infection) and autopsy-derived tissues from individuals who died while on suppressive ART. Presently, these research areas remain virtually unexplored due to the difficulty in finding subjects that fit these criteria. However, we have identified and obtained tissues from several individuals meeting our criteria and have access to many more autopsies of individuals forming the control group that have died of AIDS while not on ART. Here we propose to systematically explore the entire body for latent viral reservoirs and sanctuaries of active viral replication during ART. We propose to derive HIV-1 RNA and DNA sequences from GALT during suppressive ART, and autopsy tissues from individuals who died while on or not on suppressive ART, appraise the sequences for features of viral reservoirs, and identify tissues with features of sanctuaries of ongoing viral replication. PUBLIC HEALTH RELEVANCE: The indefinite persistence of virus reservoirs, with the capacity to fuel virus rebound, is the major obstacle to eliminating HIV infection. Identifying reservoirs of HIV-1 and sanctuaries where HIV-1 replication is not completely suppressed by antiretroviral (ARV) therapy (ART) will facilitate the exploration of new approaches to improve treatment outcomes, and will be critical to future efforts to eliminate HIV-1 from the body. Here we propose to systematically explore the entire body for latent viral reservoirs and sanctuaries of active viral replication during ART.