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 神经毒力与巨噬细胞/小胶质细胞 (MG) 的病毒趋向性有关； MG 代表存在于中枢神经系统 (CNS) 中的巨噬细胞谱系的骨髓来源细胞。在 Watry 等人进行的一系列研究中。 (1995)，通过 SIVmac251 通过受感染的 MG 连续传代，分离出一种神经病性猿猴免疫缺陷病毒 (SIV) 株；经过 3 次这样的传代后，子代 SIVmac182 在几个月内可靠地在恒河猴 (RM) 中诱导 CNS 疾病。基于这些数据，我们假设灵长类慢病毒穿过血脑屏障并在重症肌无力中有效复制的能力代表了慢病毒神经毒力的标志。我们进一步假设，已在体外适应人类 MG 的 HIV-1 包膜基因将赋予编码此类 env 基因的猿人免疫缺陷病毒（SHIV）神经毒力。我们试图在 RM 模型中检验这些假设。 迄今为止，尚未描述任何适应 MG 的 SHIV 毒株。我们已将初级、MG 适应的 R5 HIV-1 毒株的 env 插入到我们独特的 SHIV 主链中，该主链在长末端重复序列 (LTR) 中包含额外的 NF-kB 位点，从而增强了病毒复制能力和对肿瘤坏死因子 (TNF) 的反应性。 ）-？ HIV-1 env 的来源是在急性感染期间分离的菌株，然后通过培养的人类 MG 连续传代，产生 R5 子代病毒，该病毒在 MG 中生长得异常良好，并获得诱导合胞体的能力。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) 中复制良好，在 RM PBMC 中保持独特的 R5 趋向性和高融合性，并在 MG 中诱导合胞体。值得注意的是，这种新的 SHIV 在 RM 中复制得非常好，它诱导了血浆病毒 RNA 峰值峰值，并且可以在接种后 1 周内在脑脊液 (CSF) 中检测到。我们的总体目标是在 RM 中使用我们的新型 R5 SHIV 来： 目标 1：测试这样的假设：HIV-1 Env 的强 MG 向性将在感染了编码相应 env 基因的 SHIV 的 RM 中转化为神经毒力。 目标 2：通过在恒河猴中连续传代受感染的 MG 来增加 SHIV 神经毒力。将前瞻性地跟踪感染 SHIV 的动物的病毒、血液学和神经学参数。这些 R21 研究将通过建立 HIV-1 Env MG 向性和神经毒力之间的直接联系，同时建立一种新颖的、生物学相关的灵长类动物模型，为 HIV 相关痴呆的机制提供新的见解。公共健康相关性：1 型人类免疫缺陷病毒 (HIV-1) 不仅会破坏人体的免疫系统，还会攻击大脑并造成严重的神经损伤。大多数侵入大脑的 HIV-1 病毒株通过一种名为 CCR5 的受体分子进入细胞，并在小胶质细胞中有效增殖。这些特殊细胞存在于大脑中，与巨噬细胞有关。我们通过插入 HIV-1 毒株的包膜基因，从猴艾滋病病毒中产生了一种杂交病毒，该毒株已适应在小胶质细胞中有效生长。使用这种新病毒作为研究工具，我们试图研究 HIV-1 包膜基因在灵长类动物模型中引起神经损伤的作用。我们的工作将为制定更好的治疗策略提供重要信息，旨在保护大脑免受 HIV-1 攻击。