Identification and eradication of HIV tissue reservoirs in a relevant animal mode

描述(申请人提供)：虽然艾滋病毒感染的长期无进展患者和接受高效抗逆转录病毒治疗的患者在血液中检测不到病毒，但已知病毒在基本上所有艾滋病毒感染患者的整个感染过程中都持续存在于未知的组织储备库(S)。已经提出了一些组织储存库来容纳HIV-1。然而，新的证据表明，肠道相关淋巴组织(GALT)是病毒持续存在和储存的主要部位。然而，技术挑战和风险限制了在充分确定艾滋病毒感染患者的组织储存库方面的进展。这些原因要求使用动物模型对组织储存库进行评估。此前，我们使用SIV感染的中国恒河猴(CH Rh)建立了一个非人类灵长类动物模型。我们发现，与其他非人类灵长类动物模型相比，SIVmac239在CHRh中的感染更接近于HIV-1感染。此外，约1/3的CH Rh能自发控制SIV感染，尽管组织中仍有病毒残留。最近，我们发现CHRh是HAART下研究水库的首选动物模型。最近将整合酶抑制剂添加到HIV治疗武器库中，当与标准的HAART结合时，显示出显着的抑制作用。此外，我们还发现，当用包括整合酶抑制剂的抗SIV方案治疗CH Rh时，可以最大限度地抑制血浆中的病毒(&lt；30拷贝/毫升)，这一壮举在非人类灵长类动物模型中尚未实现。我们现在有了一个独特的模型，可以用来检验我们的假设，即在自发的病毒控制者或HAART中，肠道是SIV的最大储存库，并确定新的治疗策略是否最终导致从这些储存库中根除病毒。在这里，我们建议：目的1.确定HAART上感染SIV的猕猴和成为精英控制者的猕猴中的细胞和组织储存库。使用一种新的RT抑制剂和整合酶抑制剂的组合，我们可以将治疗后的NP血浆中的病毒血症降低到10RNA拷贝/毫升。我们将量化和比较肠道、骨髓、脾和其他组织中的细胞和组织储存库，以确定主要的储存库，我们还将分析HAART、非HAART和LTNP上NP之间的病毒突变和进化，以检查它们与病毒储存库和联合抗逆转录病毒治疗的关系。目的2.评估HAART联合补充方案或组织靶向策略在早期SIV感染中从受感染宿主中根除病毒的可能性。针对HIV感染的新的抗病毒策略不断涌现，我们将在SIV-猕猴模型中纳入一些最新的抗病毒方案。例如，我们目前正在猕猴身上测试融合抑制剂，这些药物可能与上述治疗相结合，试图量化多种药物组合下猕猴细胞和组织储存库的腐烂情况。此外，我们可以设计特定的策略，利用在Aim1中获得的知识来靶向特定的位置，以成功根除病毒。拟议的研究将为艾滋病毒持续和潜伏机制提供新的见解，并评估根除是否可能是一个可行的目标。最后，这些数据可能为新的治疗或疫苗方法确定有用的靶点。)

DESCRIPTION (provided by applicant): Although HIV-infected long-term nonprogressors (LTNP) and those on highly active antiretroviral therapy (HAART) have undetectable virus in blood; it is known that virus persists in yet unknown tissue reservoir(s) in essentially all HIV-infected patients throughout the course of infection. A number of tissue reservoirs have been proposed to harbor HIV-1. However, emerging evidence indicates gut-associated lymphoid tissue (GALT) is the major site of viral persistence and reservoir. However, technical challenges and risks limit progress in adequately defining tissue reservoirs in HIV-infected patients. These reasons mandate an assessment of tissue reservoirs using an animal model. Previously, we established a nonhuman primate model using SIV- infected rhesus macaques of Chinese origin (Ch Rh). We found that SIVmac239 infection in Ch Rh more closely mimics HIV-1 infection than other nonhuman primate models. Further, approximately 1/3 of Ch Rh spontaneously control SIV infection despite residual virus in tissues. Recently, we have discovered that the Ch Rh is the premier animal model for studying reservoirs under HAART. The recent addition of integrase inhibitors to the HIV therapeutic arsenal has demonstrated remarkable suppression when combined with standard HAART. Furthermore, we have found that Ch Rh, when treated with anti-SIV regimens that include integrase inhibitors have maximal suppression (< 30 copies/ml) virus in plasma, a feat that had yet to be achieved in a nonhuman primate model. We now have a unique model that can be utilized to test our hypothesis that the intestinal tract is the greatest reservoir for SIV in spontaneous viral controllers or on HAART, and determine whether novel strategies of treatment lead to eventually eradicate virus from these reservoirs. Here we propose: Aim 1. Identify the cellular and tissue reservoirs in SIV-infected macaques on HAART and those that become elite controllers. Using a novel combination of RT inhibitors and integrase inhibitors we will reduce viremia to <10 RNA copies/ml in plasma of treated NP. We will quantify and compare cellular and tissue reservoirs in the intestinal tract, bone marrow, spleen, and other tissues to determine the major reservoirs, we will also analyze viral mutations and evolution among NP on HAART, without HAART and LTNP to examine their association with viral reservoir and combination antiretroviral therapy. Aim 2. Assess the potential for eradication of virus from infected hosts by combining HAART with supplemental regimens or tissue-targeted strategies in early SIV infection. Novel antiviral strategies continue to emerge for HIV infection, and we will incorporate some of the latest antiviral regimens in the SIV-macaque model. For example, we are currently testing fusion inhibitors in macaques, and these may be combined with the above therapy in an attempt to quantify the decay of cell and tissue reservoirs in macaques on multiple drug combinations. Further, we may design specific strategies to target specific sites using the knowledge gained in Aim1 to successful viral eradication. The proposed research will provide new insights into the mechanisms of HIV persistence and latency, and assess whether eradication may be a feasible goal. Finally, these data may identify useful targets for novel therapeutic or vaccine approaches. )