Toward control of HIV neuropathogenesis by innate immunity

 描述（由申请人提供）：HIV感染的抗逆转录病毒治疗保留了免疫功能，但允许约50%的治疗患者发生神经认知障碍（NCI）。我们提出了一种预防或治疗HIV相关NCI的新途径。本申请旨在检验以下假设：单核细胞中先天性抗病毒应答的诱导可以在两个水平上减轻大脑中的HIV感染：通过防止HIV储库的建立和通过中断导致NCI的神经致病性基因产物的表达。单核细胞是HIV在大脑中的主要目标;它们介导HIV进入大脑，但它们可以启动先天免疫反应，阻止HIV感染。体内I型干扰素应答减少嵌合HIV，EcoHIV感染小鼠的神经发病机制;它们限制HIV在人类和EcoHIV感染小鼠中的传播，并可以控制猕猴中的SIV感染。我们拟利用EcoHIV感染常规和各种基因敲除小鼠及其诱导NCI，来明确小鼠巨噬细胞对HIV的保护性先天免疫应答的基础，以预防HIV在脑内的感染或其相关的脑疾病。具体目标是：1）测试Toll样受体（TLR）2、3、7/8和9的配体单独和组合用于预防EcoHIV转运至脑; HIV在脑中的储存库和NCI的逆转; 3）确定宿主的关键抗病毒决定因素 通过在特定细胞类型中使用基因表达平台、多重技术、生物信息学和荧光显微镜分析编码和调节RNA和蛋白质，在动物模型中通过TLR配体预防或治疗HAND期间的反应。方法将包括鼻内施用TLR配体、用于病毒DNA的灵敏测量的数字液滴PCR、行为和学习的恐惧条件试验、通过多重试验和显微镜检查测定巨噬细胞和脑中的抗病毒蛋白，以及通过NanoString技术进行的编码和microRNA表达的测定，以及对预防或控制EcoHIV的多种TLR配体的应答所共有的抗病毒基因或miRNA的鉴定感染NCI。如果成功，这些研究将描述新的方法和药物，用于进一步的临床开发，以治疗HIV感染者的NCI并保护脑功能。

 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.