Alcohol's Effects on HIV-1 and the Blood: Brain Barrier

酒精对 HIV-1 和血液的影响：脑屏障

• 批准号: 6753450
• 负责人: ROGER J POMERANTZ
• 金额: $ 15.7万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6753450
• 关键词: AIDS dementia complex; HIV infections; annexins; antiAIDS agent; antiviral agents; apoptosis; astrocytes; blood brain barrier; cell free system; chemokine receptor; clinical research; ethanol; human fetus tissue; human immunodeficiency virus 1; human subject; macrophage; membrane permeability; molecular pathology; neuropathology; patient oriented research; pharmacology; physical chemical interaction; protein structure function; recombinant proteins; tissue /cell culture; vascular endothelium; virus protein

DESCRIPTION (provided by applicant): Infection of the CNS of certain individuals by HIV-1 may lead to significant cognitive and motor dysfunction, entitled HIV-1-induced encephalopathy. In addition, a number of cofactors including ethanol (Etoh) ingestion may alter penetration of HIV-1 into the CNS through the blood:brain barrier (EBB). Under-standing these cofactors involved with penetration of HIV-1 into the CNS reservoir is of critical importance now in the era of HAART. Our laboratories have recently developed an in vitro BBB system comprised totally of human CNS cell-types. We have demonstrated that various viral proteins, especially HIV-1 Vpr, has profound effects on CNS-based cells leading to apoptotic death. Utilizing complementary approaches, the effects of Etoh on HIV-1 infection of the BBB and viral penetration of the CNS will be -1 virus in toto or specific dissected . It is hypothesized that Etoh may have significant effects with the HIV HIV-1 proteins in inducing the disruption of the BBB, possibly through apoptosis, as well as increasing infection of BBB-constituting cells, including microvascular endothelial cells (MVECS) and astrocytes. In Specific Aim 1, the cellular and molecular effects of Etoh on HIV-1 infection, with and without anti-retroviral agents, of critical CNS cells in the BBB will be approached. Specific viral strains in cell-free and cell- associated (e.g, infected macrophages) forms will be utilized. In Specific Aim U, effects of Etoh with selected recombinant HIV-1 proteins on disruption of this barrier will be mechanistically analyzed. Potential disruption of the BBB by Etoh's effects with MV-1 proteins will be assessed (as for S.A.1) using complementary methods, including trans-endothelial electrical resistance (TEER) and ZOI tight junction protein alterations. Etoh's effects with select recombinant Viral proteins on apoptosis of specific BBB cells, will be analyzed via TUNEL, Annexin V assays and ion channel measurements. These effects will also be probed using mutant viruses lacking specific viral regulatory and structural genes, including Vpr, Nef, gpl2O and Tat. HIV- I vector systems to express these specific proteins intracellularly will be utilized with Etoh. Finally, alterations of specific chemokine receptors on BBB cells, especially the CNS-specific chemokine receptor APJ, will be analyzed in the setting of Etoh. Effects on chemokine receptor expression, down- regulation, and intracellular calcium fluxes after binding to relevant cognate chemokines and gpl2O will be analyzed. With these inter-related specific aims, it is proposed that a series of new technologies will be combined towards determining the precise effects of Etoh on inducing alterations in the BBB with HW-l I infection of these cells.

描述（由申请人提供）： HIV-1对某些个体的中枢神经系统的感染可能导致严重的认知和运动功能障碍，标题为HIV-1诱导的脑病。此外，包括乙醇（ETOH）在内的许多辅助因子可能会通过血液屏障（EBB）改变HIV-1进入中枢神经系统的渗透。在HAART时代，与HIV-1渗透到中枢神经系统储层中有关的这些辅助因子与渗透到中枢神经系统储层中至关重要。我们的实验室最近开发了一个完全由人类CNS细胞类型组成的体外BBB系统。我们已经证明，各种病毒蛋白，尤其是HIV-1 VPR，对基于CNS的细胞产生深远影响，导致凋亡死亡。利用互补方法，ETOH对BBB的HIV -1感染和CNS病毒渗透的影响将为-1个病毒或特异性解剖。 假设ETOH可能会对HIV HIV-1蛋白具有显着影响，从而诱导BBB的破坏，可能通过凋亡，以及增加BBB构造细胞的感染，包括微血管内皮细胞（MVEC）和星形镜。在特定的目标1中，将接近ETOH对HIV-1感染的细胞和分子作用，具有和没有抗逆转录病毒药物的BBB中关键CNS细胞的细胞和分子作用。将使用无细胞和相关细胞（例如感染巨噬细胞）形式的特定病毒菌株。在特定的目标u中，将通过机械分析ETOH对选定的重组HIV-1蛋白对该屏障的破坏的影响。通过ETOH对MV-1蛋白的作用对BBB的潜在破坏（如S.A.1），使用互补方法，包括跨内皮电阻（TEER）和ZOI紧密连接蛋白改变。 ETOH对特定BBB细胞凋亡的精选重组病毒蛋白的影响将通过TUNEL，ANNEXIN V分析和离子通道测量值分析。这些作用还将使用缺乏特定病毒调节和结构基因（包括VPR，NEF，GPL2O和TAT）的突变病毒进行探测。 HIV-i载体系统以EtOH利用细胞内表达这些特定蛋白质。最后，将在ETOH的情况下分析BBB细胞上特定趋化因子受体的改变，尤其是CNS特异性趋化因子受体APJ。将分析与相关的同源趋化因子和GPL2O结合后对趋化因子受体表达，下调和细胞内钙通量的影响。通过这些相互关联的特定目的，建议将一系列新技术结合起来，以确定EtOH对诱导BBB诱导这些细胞感染的BBB改变的确切影响。