Project 4: Intracellular mechanism of virus persistence in astrocytes/macrophages

项目4：病毒在星形胶质细胞/巨噬细胞中持续存在的细胞内机制

• 批准号: 6983327
• 负责人: AVINDRA NATH
• 金额: $ 22.74万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-12-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6983327
• 关键词: AIDS /HIV neuropathy; AIDS therapy; HIV infections; antiAIDS agent; astrocytes; brain; cell line; combination chemotherapy; embryo /fetus tissue /cell culture; human fetus tissue; human immunodeficiency virus; immunogenetics; interferon inducers; latent virus infection; macrophage; nervous system infection; neurotropic virus; transfection; virus genetics; virus protein; virus replication

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.

大脑可能是人类免疫缺陷病毒（HIV）的重要储存库。很明显，艾滋病毒会感染大脑中入侵的巨噬细胞、驻留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。目前还不清楚抗逆转录病毒治疗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。因此，我们建议确定病毒进入后这些细胞类型中维持病毒的机制，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。我们假设大脑中病毒储存库的建立涉及特定的抗病毒基因，这些基因 巨噬细胞和星形胶质细胞中的病毒受到不同程度的调节，一旦建立储存库，抗逆转录病毒治疗对病毒在这些细胞中的持续存在影响很小或没有影响。我们将结合使用体外人脑培养物和血液来源的巨噬细胞来实现以下目标。 这些研究将补充项目#1中的研究，项目#1将研究体内和离体巨噬细胞中的病毒持久性，项目#2和项目#3将确定体内抗逆转录病毒治疗对大脑的影响。我们实验室的初步数据表明，HIV 的 Tat 蛋白调节多种干扰素诱导抗病毒基因的表达。我们已经产生了几种新型细胞系和病毒构建体，这将使我们能够解决与大脑中病毒库相关的这些问题。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. 确定 HIV 对星形胶质细胞的影响 巨噬细胞抗病毒基因、干扰素刺激基因 20 (ISG 20) 和寡腺苷酸合成酶 (OAS-1)。图 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。