Effects of Opioids on SIV Reservoirs in Brain Macrophages of Rhesus Macaques

 描述：艾滋病毒感染和鸦片类药物成瘾是一个新出现的和日益严重的问题，部分原因是HAART提供了更长的预期寿命。尽管有效的抗逆转录病毒疗法(ART)和延长寿命，阿片类药物的使用加剧了艾滋病毒感染患者的神经异常。我们的长期目标是改善或逆转神经异常的发展，尽管药物成瘾的艾滋病毒感染者出现了这种异常。R21/R33建议的目的是验证和确定血管周围巨噬细胞(PVM)在抗逆转录病毒治疗后对SIV感染猕猴中枢神经系统中病毒库的贡献，并测试阿片成瘾是否改变免疫细胞亚群(包括PVM)的功能，从而为病毒库提供燃料，并加剧SIV感染的发病。我们的中心假设是，长寿命的PVM不仅是主要的SIV储存库，而且还影响中枢神经系统其他细胞亚群的病毒储存库的建立，如阿片成瘾猕猴的小胶质细胞和星形胶质细胞。为了验证这一点，我们提出，在体内耗尽长寿命的PVM将减少或消除病毒库，从而改善神经病理和神经炎症的进展。我们假设的基本原理是：(I)在感染hiv和成瘾的人类中同时观察到的依赖吗啡的siv感染的猕猴表现出神经异常的恶化，(Ii)与hiv感染的人类相比，在猕猴中可以更详细地和在更可控的实验条件下检测siv的储存点，以及(Iii)我们建立了一种安全的方案来消除体内的PVM，这将使我们能够直接证明它们在病毒储存库中的作用。 和神经发病机制。我们在此应用的第一阶段(R21)提出了两个目标：目的1.确定吗啡对接受抗逆转录病毒治疗的SIV感染猕猴中枢神经系统病毒库的大小和宿主细胞范围的影响。我们的工作假设是，在ART治疗开始时，经吗啡治疗的SIV感染猕猴将在PVM中形成更高的病毒储存库水平和更高的单核细胞周转率(指示疾病进展)。目的2.明确PVM对阿片依赖型SIV感染猕猴中枢神经系统病毒库的特殊贡献。我们的工作假设是，在体内耗尽PVM将减少或消除接受吗啡治疗的SIV感染猕猴接受抑制ART的脑病毒储存库。为了测试这一点，感染SIV的猕猴将在达到病毒设定点后开始抗逆转录病毒治疗。然后，PVM将通过鞘内双膦酸脂质体(BP)治疗选择性地被耗尽，该治疗有望通过招募新的细胞来诱导再生。这一方法还将揭示其他可能导致SIV储存库的细胞类型的大小，如小胶质细胞和星形胶质细胞。这些结果将显示在经吗啡治疗的SIV感染猕猴大脑中SIV储存库的细胞位置(S)和大小。具体地说，这些结果将为体内PVM的耗尽及其对病毒库的贡献提供概念验证，从而 导致测试针对PVM的治疗策略，以减少疾病进展。这将为第二阶段目标3(R33)的研究奠定基础，以证实感染后早期靶向PVM的可能性，以及ART将减少或改善吗啡治疗的SIV感染猕猴中枢神经系统发病的恶化。目的3.确定体内早期耗尽PVM是否能阻止阿片依赖型SIV感染的猕猴在接受抑制ART治疗时的神经病变进展。我们的工作假设是，PVM作为SIV储集层和/或促进其他宿主细胞中储集层的开发。在这里，我们将确定在启动ART后早期体内消除PVM是否可以控制或减缓慢性吗啡治疗的SIV感染猕猴中枢神经系统疾病的进展。如果成功，这项研究将导致开发新的治疗策略，有效地针对脑部持续的艾滋病毒感染和逆转手的发育。

 DESCRIPTION: HIV infection in combination with opiate drug addiction is an emerging and growing problem, partly due to a longer life expectancy afforded by HAART. Despite effective anti-retroviral therapy (ART) and extended longevity, opiate use exacerbates neurological abnormalities in HIV-infected patients. Our long-term goal is to ameliorate or reverse development of neurological abnormalities that arise despite ART in drug-addicted HIV-infected individuals. The purpose of this R21/R33 proposal is to verify and determine the contribution of perivascular macrophages (PVM) to the virus reservoir in the CNS of SIV-infected macaques following ART and to test if opiate addiction modifies the function of immune cell subsets, including PVM, to fuel the virus reservoir and exacerbate pathogenesis of SIV infection. Our central hypothesis is that long-lived PVM not only serve as major SIV reservoirs but also influence the establishment of virus reservoirs in other cells subsets of the CNS such as microglia and astrocytes in opioid-addicted macaques. To test this, we propose that in vivo depletion of long-lived PVM will reduce or eliminate the virus reservoir and consequently ameliorate the progression of neuropathology and neuroinflammation. The rationale for our hypothesis is that, (i) morphine- dependent SIV-infected macaques display exacerbation of neurological abnormalities observed in parallel in HIV-infected and addicted humans, (ii) reservoir sites of SIV can be examined in finer detail and under more controlled experimental conditions in macaques than in HIV-infected humans, and (iii) we established a safe protocol to eliminate PVM in vivo that will enable us to directly demonstrate their role in the virus reservoir and neuropathogenesis. We propose two aims in phase I (R21) of this application: Aim 1. Determine the effect of morphine on the size and host cell range of the CNS virus reservoir in SIV-infected macaques undergoing ART. Our working hypothesis is that morphine- treated SIV-infected macaques will develop a higher virus reservoir level in PVM and a higher monocyte turnover (indicative of disease progression) than in the SIV-infected-only group at the initiation of ART treatment. Aim 2. Define the specific contribution of PVM to the CNS viral reservoir in opioid-dependent SIV-infected macaques undergoing suppressive ART. Our working hypothesis is that in vivo depletion of PVM will reduce or eliminate the brain virus reservoir in morphine-treated SIV-infected macaques undergoing suppressive ART. To test this, SIV-infected macaques will begin ART after reaching virus set point. The PVM then will be selectively depleted via intrathecal liposomal bisphosphonate (BP) treatment that is expected to induce repopulation by recruiting fresh cells. This approach will also bring to light the magnitude of other cell types that may contribute to the SIV reservoir, such as microglia and astrocytes. These results will demonstrate cellular site(s) and magnitude of SIV reservoirs in the brain of morphine-treated SIV-infected rhesus macaques. Specifically, the outcomes will provide proof-of-concept for the in vivo depletion of PVM and their contribution to the virus reservoir that will lead to testing a treatment strategy targeting PVM to reduce disease progression. This will set the foundation for stud- ies in aim 3 of phase II (R33) to corroborate if targeting PVM earlier after infection and ART will re- duce or ameliorate exacerbation of CNS pathogenesis in morphine-treated SIV-infected macaques. Aim 3. Determine if early in vivo depletion of PVM prevents progression of neuropathogesis in opioid-dependent SIV-infected macaques undergoing suppressive ART. Our working hypothesis is that PVM serve as SIV reservoirs and/or promote development of reservoirs in other host cells. Here, we will determine if in vivo elimination of PVM early after initiating ART will control or re- duce progression of CNS pathogenesis in SIV-infected macaques with chronic morphine treatment. If successful, this study will lead to developing novel therapeutic strategies to effectively target persistent HIV infection of the brain and reverse HAND development.