HIV-1 Env Microglial Tropism and Neurovirulence

HIV-1 Env 小胶质细胞趋性和神经毒力

• 批准号: 7622988
• 负责人: Ruth Margrit Ruprecht
• 金额: $ 21.58万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7622988
• 关键词: AIDS Dementia Complex; Acute; Address; Animals; Appendix; Autologous; Blood - brain barrier anatomy; Bone Marrow; Brain; CCR5 gene; CD8-Positive T-Lymphocytes; Cells; Central Nervous System Diseases; Cerebrospinal Fluid; Clinical; Cloning; Cognitive; Data; Development; Disease; Disease Progression; Encephalitis; Epidemic; Foundations; Future; Genes; Giant Cells; Goals; HIV; HIV-1; Histopathology; Human; Hybrids; Immune system; Immunohistochemistry; Impairment; In Situ Hybridization; In Vitro; Infection; Invaded; Life; Link; Long Terminal Repeats; Macaca mulatta; Microglia; Minor; Modeling; Monkeys; Monoclonal Antibodies; NF-kappa B; Nervous System Trauma; Neuraxis; Neurologic; Numbers; Patients; Peripheral Blood Lymphocyte; Peripheral Blood Mononuclear Cell; Plasma; Play; Primate Lentiviruses; Primates; Proliferating; Public Health; Research; Role; SIV; Serial Passage; Series; Site; Source; Stroke; Subfamily lentivirinae; Terminal Repeat Sequences; Testing; Tissue Donors; Translating; Tropism; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Vertebral column; Viral; Virus; Week; Work; base; brain tissue; cytotoxic; env Genes; human TNF protein; in vivo; insight; macrophage; monocyte; motor disorder; nervous system disorder; neuropsychological; neurovirulence; novel; receptor; simian human immunodeficiency virus; tool; viral RNA

DESCRIPTION (provided by applicant): HIV neurovirulence has been linked to viral tropism for macrophages/microglia (MG); MG represent bone marrow-derived cells of macrophage lineage residing in the central nervous system (CNS). In a series of studies performed by Watry et al. (1995), a neuropathogenic simian immunodeficiency virus (SIV) strain was isolated by serial passage of SIVmac251 through infected MG; after 3 such passages, the progeny SIVmac182 reliably induced CNS disease in rhesus monkeys (RM) within a few months. Based upon these data, we postulate that the ability of a primate lentivirus to cross the blood brain barrier and to replicate efficiently in MG represents a hallmark of lentiviral neurovirulence. We further postulate that the envelope gene of HIV-1 that has been adapted in vitro to human MG will confer neurovirulence to a simian-human immunodeficiency virus (SHIV) encoding such an env gene. We seek to test these hypotheses in the RM model. To date, no MG-adapted SHIV strain has been described. We have inserted env of a primary, MG- adapted R5 HIV-1 strain into our unique SHIV backbone that contains extra NF-kB sites in the long terminal repeats (LTRs), which boosts viral replicative capacity and responsiveness to tumor necrosis factor (TNF)-?. The source for HIV-1 env was a strain isolated during acute infection and then serially passaged through cultured human MG, yielding an R5 progeny virus that grew exceptionally well in MG and gained the ability to induce syncytia. Our novel SHIV construct, SHIV-Bo159N4, replicated well in peripheral blood mononuclear cells (PBMC) of all RM donors tested, maintained the exclusive R5 tropism and high fusogenicity in RM PBMC and induced syncytia in MG. Remarkably, this new SHIV replicated very well in RM, where it induced high peak plasma viral RNA loads and could be detected in cerebrospinal fluid (CSF) within 1 week post- inoculation. Our overall goal is use our novel R5 SHIV in RM to: Aim 1: Test the hypothesis that strong MG tropism of HIV-1 Env will translate into neurovirulence in RM infected with a SHIV encoding the corresponding env gene. Aim 2: Increase SHIV neurovirulence by serial passage through infected MG in rhesus macaques. SHIV-infected animals will be followed prospectively for viral, hematological and neurological parameters. These R21 studies will give new insights into mechanisms of HIV-associated dementia by establishing a direct link between HIV-1 Env MG tropism and neurovirulence while establishing a novel, biologically relevant primate model. PUBLIC HEALTH RELEVANCE: Human immunodeficiency virus type 1 (HIV-1) not only destroys the body's immune system, but can also attack the brain and cause severe neurological damage. Most HIV-1 strains that invade the brain enter cells through a receptor molecule called CCR5 and proliferate efficiently in microglia. These specialized cells reside in the brain and are related to macrophages. We have generated a hybrid virus from the monkey AIDS virus by inserting the envelope gene of an HIV-1 strain that has been adapted to grow efficiently in microglia. Using this new virus as a research tool, we seek to study the role that the HIV-1 envelope gene plays in causing neurological damage in primate models. Our work will provide important information for developing a better treatment strategies aimed at protecting the brain from HIV-1 attack.

描述（由申请人提供）：HIV神经动力学与巨噬细胞/小胶质细胞（MG）的病毒疗法有关； MG代表中枢神经系统（CNS）中巨噬细胞谱系的骨髓衍生细胞。在Watry等人进行的一系列研究中。 （1995年），通过通过感染的MG的SIVMAC251的串行传递来分离神经病的邻肌免疫缺陷病毒（SIV）菌株；经过3个这样的通道后，后代SIVMAC182在几个月内可靠地诱发了恒河猴（RM）中的CNS疾病。基于这些数据，我们假设灵长类动病毒越过血脑屏障和在MG中有效复制的能力代表了慢病毒神经动力毒素的标志。我们进一步假设，在体外适应了人类MG的HIV-1的包膜基因将赋予神经动力学与编码这种ENV基因的猿猴 - 人类免疫缺陷病毒（SHIV）。我们试图在RM模型中检验这些假设。 迄今为止，尚未描述MG适应的SHIV菌株。我们已经将ENV插入了一个主要的MG-Spappated R5 HIV-1菌株中的独特的SHIV主链，该主链在长期重复序列（LTRS）中包含额外的NF-KB位点（LTRS），从而提高了病毒复制能力和对肿瘤坏死因子的响应能力（TNF） ） - ？。 HIV-1 Env的来源是在急性感染过程中分离出来的菌株，然后连续通过培养的人Mg，产生了R5后代病毒，在MG中生长得非常好，并获得了诱导合胞体的能力。我们的新型SHIV构建体Shiv-BO159N4在所有测试的RM供体的外周血单核细胞（PBMC）中很好地复制了，维持RM PBMC的独家R5 Tropism和高融合性，并诱导MG中的合成症。值得注意的是，这种新的SHIV在RM中很好地复制了，它诱导了高峰血浆病毒RNA负载，并且可以在接种后1周内在脑脊液（CSF）中检测到。我们的总体目标是将我们的新型R5 SHIV在RM中使用：AIM 1：测试假设HIV-1 Env的强烈MG TROPISM将转化为RM中的神经动力，感染了编码相应的Env Gene的SHIV。 AIM 2：通过恒河猕猴中感染的毫克连续传递来增加SHIV神经动力。感染的动物将被前瞻性地遵循病毒，血液学和神经系统参数。这些R21研究将通过在建立一种新颖的生物学相关的灵长类动物模型的同时建立HIV-1 Env MG Tropism和神经动力毒气之间的直接联系，从而为与HIV相关痴呆的机制提供新的见解。公共卫生相关性：人类免疫缺陷病毒1型（HIV-1）不仅破坏了人体的免疫系统，而且还会攻击大脑并造成严重的神经系统损害。大多数HIV-1菌株通过称为CCR5的受体分子进入细胞并在小胶质细胞中有效增殖。这些专门的细胞驻留在大脑中，与巨噬细胞有关。我们通过插入已适应以在小胶质细胞中有效生长的HIV-1菌株的包膜基因来从猴子艾滋病病毒中产生混合病毒。我们使用这种新病毒作为研究工具，试图研究HIV-1信封基因在灵长类动物模型中造成神经系统损害中起作用的作用。我们的工作将提供重要的信息，以制定旨在保护大脑免受HIV-1攻击的更好治疗策略。