NeuroAIDS Therapy in the EcoHIV Mouse Model

描述（由申请人提供）：对HIV-1感染的高效抗逆转录病毒治疗（HAART）在大多数依从性患者中有效地减少了全身病毒负担并改善了免疫功能。然而，hiv -1相关的痴呆和其他神经系统疾病仍然存在。长寿命巨噬细胞和脑细胞中的病毒储存库对HAART不敏感，并且某些药物无法进入大脑。本应用程序旨在通过一种新的嵌合病毒EcoHIV/NDK感染免疫功能正常小鼠的HIV-1模型，实验研究抗病毒药物对HIV-1在大脑中的复制和发病机制的影响。这种病毒编码除gp120外的所有HIV-1蛋白，因此保持对HAART药物的敏感性。我们最近报道了zalcitabine和abacavir阻断小鼠EcoHIV/NDK感染，并在该应用中显示奈韦拉平和蛋白酶抑制剂沙奎那韦显著减少病毒在小鼠中的传播。利用实时聚合酶链反应（real-time PCR）对大量受感染动物和多个组织中的病毒进行监测，可以对药物疗效进行定量测试，并通过分析细胞转录本，研究我们模型中的脑细胞功能障碍。在这一发展计划中，我们建议1)测试抗逆转录病毒药物预防成年和年轻小鼠的急性外周和中枢神经系统感染EcoHIV/NDK的功效，2)测试抗逆转录病毒药物减少年轻小鼠外周组织和大脑中EcoHIV/NDK感染及其相应的神经发病机制的功效。实时荧光定量PCR检测淋巴细胞、巨噬细胞和大脑中的病毒负荷以及脑细胞基因表达的激活情况，免疫细胞化学检测小胶质细胞的激活情况。该项目直接研究难以进入大脑的药物是否能阻止HIV-1和脑部疾病的神经系统储存库的建立，大脑中的病毒是否对HAART药物敏感，以及抑制正在进行的病毒复制在多大程度上耗尽脑细胞、巨噬细胞和骨髓中的病毒储存库并改善脑部疾病。该发展计划旨在指导未来在该动物模型中的转化研究，以筛选或开发新的药物，以预防hiv -1相关的神经发病机制。HIV-1感染会导致免疫缺陷，通常还会导致神经系统紊乱，导致记忆丧失、思维混乱和运动控制不良。该应用建议使用实验性感染HIV-1的小鼠和目前用于人类的抗病毒药物来确定它们是否阻断小鼠大脑中的感染和疾病。此外，该应用程序还将测试感染后给老鼠服用的药物是否真的能消除大脑中的病毒，这为抗病毒药物控制人类感染的神经系统疾病带来了希望。

DESCRIPTION (provided by applicant): Highly active antiretroviral therapy (HAART) for HIV-1 infection effectively reduces systemic virus burden and improves immune function in most compliant patients. Nevertheless, HIV-1-associated dementia and other nervous system disorders persist. Viral reservoirs in long-lived macrophages and brain cells are not sensitive to HAART and the brain is inaccessible to certain drugs. This application is designed to investigate the effects of antiviral drugs on HIV-1 replication and pathogenesis in the brain experimentally using a new model of HIV-1 infection of immunocompetent mice by the chimeric virus, EcoHIV/NDK. This virus encodes all the proteins of HIV-1, except gp120, and thus preserves sensitivity to HAART agents. We recently reported that zalcitabine and abacavir block EcoHIV/NDK infection of mice and show in this application that nevirapine and the protease inhibitor saquinavir significantly reduce virus spread in mice. Access to a large number of infected animals and monitoring of virus in multiple tissues using real-time PCR permit quantitative test of drug efficacy, and through assay of cellular transcripts, brain cell dysfunction in our model. In this developmental program we propose to 1) test the efficacy of antiretroviral drugs for prophylaxis of acute peripheral and central nervous system infection of adult and young mice by EcoHIV/NDK and 2) test the efficacy of antiretroviral drugs for reduction of established EcoHIV/NDK infection in peripheral tissues and brain and consequential neuropathogenesis in young mice. Virus burden in lymphocytes, macrophages, and the brain and activation of brain cell gene expression will be determined by real-time PCR and microglial activation will be assessed by immunocytochemistry. This program directly investigates whether drugs with poor access to the brain can prevent establishment of a nervous system reservoir of HIV-1 and brain disease, whether virus resident in the brain is sensitive to HAART agents, and to what extent inhibition of ongoing virus replication depletes viral reservoirs in brain cells, macrophages, and bone marrow and ameliorates brain disease. This developmental program is proposed to guide future translational studies in this animal model to screen or develop new agents for their ability to prevent HIV-1-associated neuropathogenesis. PUBLIC HEALTH RELEVANCE HIV-1 infection causes immunodeficiency and often nervous system disorders resulting in memory lapses, confusion, and poorly controlled movements. This application proposes to use mice experimentally infected by HIV-1 and antiviral drugs now used in human beings to determine whether they block infection and disease in the mouse brain. In addition, the application will test if drugs given to mice after infection actually eliminate virus from the brain, giving promise for the ability of antiviral drugs to control the nervous system disease seen in infected human beings.