Intracellular mechanisms maintaining virus persistence in astrocytes & macrophage

大脑可能是人类免疫缺陷病毒（HIV）的重要储存库。很明显，HIV感染了入侵的巨噬细胞，常驻的小胶质细胞和星形胶质细胞。然而，病毒长时间驻留在这些细胞中的基本机制尚不清楚。抗逆转录病毒疗法是否可以控制这些细胞中的病毒复制或对大脑中的病毒库产生影响仍然是未知的。因此，我们建议确定参与维持病毒在这些细胞类型后病毒进入的机制，并确定抗逆转录病毒治疗对这些细胞中病毒生命周期的影响。我们推测，大脑中病毒储库的建立涉及特定的抗病毒基因， 在巨噬细胞和星形胶质细胞中受到不同的调节，一旦建立了储库，抗逆转录病毒治疗对这些细胞中病毒的持续存在几乎没有影响。我们将使用体外人脑培养物和血液来源的巨噬细胞的组合来实现以下目标。 这些研究将补充项目1中的研究，该项目将研究体内和体外巨噬细胞中的病毒持久性，项目2和3将确定抗逆转录病毒治疗对体内脑的影响。我们实验室的初步数据表明，HIV的达特蛋白调节几种干扰素诱导的抗病毒基因的表达。我们已经产生了几种新的细胞系和病毒构建体，这将使我们能够解决这些与大脑中病毒库相关的问题。因此，我们提出以下具体目标：1.确定抗病毒蛋白-早幼粒细胞白血病蛋白（PML）在HIV中的作用 在星形胶质细胞和巨噬细胞中复制。2.为了确定HIV在星形胶质细胞中的作用， 巨噬细胞对抗病毒基因、干扰素刺激基因-20（ISG 20）和寡腺苷酸合成酶（OAS-1）的影响。3：确定CART对星形胶质细胞和巨噬细胞中HIV复制和持久性的影响。

The brain is likely an important reservoir for the human immunodeficiency virus (HIV). It is clear that HIV infects the invading macrophages, resident microglia and astrocytes in the brain. However, the fundamental mechanisms by which the virus resides in these cells for prolonged periods of time is not understood. It also remains unknown if antiretroviral therapy can control viral replication in these cells or have an impact on the viral reservoirs in brain. We thus propose to determine mechanisms involved in maintaining the virus in these cell types post viral entry and determine the effect of antiretroviral therapy on the viral lifecycle in these cells. We hypothesize that establishment of a viral reservoir in brain involves specific antiviral genes which are differentially regulated in macrophages and astrocytes and once the reservoir is established, antiretroviral therapy will have little or no effect on the persistence of the virus in these cells. We will use a combination of in vitro human brain cultures and blood derived macrophages to address the aims below. These studies will compliment those in project #1 which will study viral persistence in macrophages in vivo and ex vivo and projects #2 and 3 that will determine the effect of antiretroviral therapy on brain in vivo. Preliminary data from our laboratory suggests that the Tat protein of HIV regulates the expression of several interferon inducible antiviral genes. We have generated several novel cell lines and viral constructs that will allow us to address these issues related to viral reservoirs in the brain. We thus propose the following specific aims: 1: To determine the role of antiviral protein, promelyocytic leukemia protein (PML) in HIV replication in astrocytes and macrophages. 2. To determine the effect of HIV in astrocytes and macrophages on antiviral genes, interferon stimulated gene-20 (ISG 20) and oligoadenylate synthetase (OAS-1). 3: To determine the effect of CART on HIV replication and persistence in astrocytes and macrophages.