Toward control of HIV neuropathogenesis by innate immunity

 描述(申请人提供)：艾滋病毒感染的抗逆转录病毒治疗保留了免疫功能，但允许大约50%的接受治疗的患者出现神经认知障碍(NCI)。我们提出了一条预防或治疗HIV相关NCI的新途径。这项应用旨在测试这一假设，即在单核细胞中诱导先天抗病毒反应可以在两个水平上减轻大脑中的艾滋病毒感染：通过防止艾滋病毒储存库的建立和通过中断导致NCI的神经致病基因产物的表达。单核细胞是艾滋病毒在大脑中的主要靶点；它们介导艾滋病毒进入大脑，但它们可以启动先天免疫反应，阻止艾滋病毒感染。在体内，I型干扰素反应减少了感染嵌合HIV的小鼠的神经致病作用；它们限制了HIV在人类和EcoHIV感染小鼠中的传播，并可以控制SIV在猕猴中的感染。我们建议利用常规和各种基因敲除小鼠的EcoHIV感染及其诱导的NCI，来确定巨噬细胞对HIV的保护性先天免疫反应的基础，以预防HIV在脑内的感染或与其相关的脑部疾病。具体目标是：1)单独和组合测试Toll样受体(TLR)2、3、7/8和9的配体，以防止EcoHIV传播到大脑；2)测试TLR配体对TLR的沉默作用 大脑中的HIV储存库和NCI的逆转；3)确定宿主的关键抗病毒决定因素 在动物模型中，通过使用基因表达平台、多重技术、生物信息学和荧光显微镜对编码和调控的RNA和蛋白质进行分析，研究TLR配体在预防或治疗手部疾病过程中的反应。方法将包括鼻腔注射TLR配体，用于敏感测量病毒DNA的数字液滴聚合酶链式反应，行为和学习的恐惧条件分析，通过多重分析和显微镜检测巨噬细胞和大脑中的抗病毒蛋白，通过纳米串技术分析编码和微型RNA的表达，并识别与多种TLR配体的反应共同的抗病毒基因或miRNA，以防止或控制NCI的EcoHIV感染。如果成功，这些研究将为进一步的临床开发描述新的方法和试剂，以治疗艾滋病毒感染者的NCI并保护大脑功能。

 DESCRIPTION (provided by applicant): Antiretroviral therapy in HIV infection preserves immune function but allows development of neurocognitive impairment (NCI) in about 50% of treated patients. We propose a new route to prevent or treat HIV-associated NCI. This application aims to test the hypothesis that induction of innate antiviral responses in monocytes can mitigate HIV infection in the brain at two levels: by preventing establishment of the HIV reservoir and by interruption of expression of neuropathogenic gene products leading to NCI. Monocytic cells are the primary target of HIV in the brain; they mediate HIV entry into the brain, but they can mount innate immune responses that block HIV infection. In vivo Type I interferon responses reduce neuropathogenesis in mice infected by chimeric HIV, EcoHIV; they restrict HIV transmission in human beings and in EcoHIV infected mice and can control SIV infection in macaques. We propose to use EcoHIV infection of conventional and various knockout mice and its induction of NCI, to define the basis of protective innate immune responses of macrophages to HIV in mice to prevent HIV infection in the brain or its associated brain disease. The Specific Aims are: 1) to test ligands to Toll-like receptors (TLR) 2, 3, 7/8, and 9 singly and in combinations for prevention of EcoHIV transit to the brain; 2) to test TLR ligands for silencing of the HIV reservoir in the brain and reversal of NCI; 3) to define key antiviral determinants of host responses during prevention or therapy of HAND by TLR ligands in an animal model by analysis of coding and regulatory RNAs and proteins using gene expression platforms, multiplex technology, bioinformatics, and fluorescence microscopy in specific cell types. Methods will include intranasal administration of TLR ligands, digital droplet PCR for sensitive measurement of viral DNA, fear conditioning assays of behavior and learning, assay of antiviral proteins in macrophages and brain by multiplex assays and microscopy, and assays of both coding and microRNA expression by NanoString technology and identification of antiviral genes or miRNA common to responses to multiple TLR ligands that prevent or control EcoHIV infection of NCI. If successful, these studies will describe novel approaches and agents for further clinical development to treat NCI in HIV infected people and to preserve brain function.