Role of gp120 in Neuropathogenesis in a New Model of HIV Infection in Mice

gp120 在小鼠 HIV 感染新模型神经发病机制中的作用

• 批准号: 7065917
• 负责人: DAVID J VOLSKY
• 金额: $ 18.16万
• 依托单位: ST. LUKE'S-ROOSEVELT INST FOR HLTH SCIS
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7065917
• 关键词: HIV infections; astrocytes; brain; infection; macrophage; model; receptor; role; tissue mosaicism

DESCRIPTION (provided by applicant): We have recently established a novel model for HIV-1 infection in healthy immunocompetent mice. Mice inoculated i.v. or i.p. with EcoHIV, a chimeric HIV-1 carrying MLV envelope that mediates HIV-1 entry into mouse cells, become persistently infected, with production of antibodies to viral proteins and presence of virus in the spleen, peritoneal macrophages, and PBL. A large proportion of inoculated animals show evidence of early virus infiltration into the brain and elevated expression of genes such as MCP-1, STAT1, and IL-1b, which are associated with inflammation in brain tissue in humans. In contrast, relatively few animals displayed brain lesions indicative of overt brain disease. Thus EcoHIV is neurotropic and neuropathogenic in mice but it does not express HIV-1 gp120 and it appears to cause attenuated, "preclinical" brain disease in mice. HIV-1 gp120 is thought to contribute to neuropathogenesis through interaction with surface receptors on macrophages, astrocytes, and neurons. Preliminary studies indicate that HIV-1 carrying native gp120 induces a wider range of inflammatory molecules in mouse astrocytes in culture than EcoHIV. We hypothesize that expression of selected gp120 segments, such as V3, by EcoHIV will facilitate HIV-1-mediated neuropathogenesis in mice. The exploratory studies proposed here will evaluate this hypothesis in two Aims. In Aim 1, we will construct a series of chimeric EcoHIV with increasingly large HIV-1 gp120 regions around V3, including those responsible for interaction with surface receptors, inserted into MLV Env of EcoHIV. The chimeras will be tested for infectivity in vitro and in vivo and for their ability to induce inflammatory proteins in mouse macrophages and astrocytes in culture compared to EcoHIV. The first such chimera we constructed, EcoHIV-V3, was found to be infectious in vivo. In Aim 2, we will compare selected gp120- expressing chimeras and EcoHIV for induction of neuropathogenesis in mice. We will determine the kinetics of virus infection and neuroinvasion in vivo, the expression of selected molecular markers of HIV-1 neuropathogenesis in the brain, and the development of neuropathology. The proposed studies may help to define the role of HIV-1 gp120 in HIV-1-mediated neuropathogenesis in mice in distinction from contribution of the viral genome lacking gp120. The studies employ an animal model, molecular, and pathological analyses of animal tissues, and molecular virology and recombinant DMA technologies. In the larger public health context, our studies will establish the utility of this animal model of HIV-1 infection in research on HIV-1 pathogenesis, including that in the central nervous system, development of HIV-1 vaccine, and convenient inexpensive testing of antiviral compounds in vivo.

描述（由申请人提供）：我们最近在健康免疫能力小鼠中建立了一种新型HIV-1感染模型。接种I.V.的小鼠或i.p.使用EcoHIV，一种携带MLV包膜的嵌合HIV-1介导HIV-1进入小鼠细胞，持续感染了病毒蛋白的抗体，并在脾，腹膜巨噬细胞和PBL中产生抗体。很大一部分接种的动物表现出早期病毒浸润到大脑中的证据，以及与MCP-1，STAT1和IL-1B等基因的表达升高，这些基因与人类脑组织的炎症有关。相反，相对较少的动物表现出脑部病变，表明明显的脑部疾病。因此，ECOHIV是小鼠的神经性和神经病性，但并未表达HIV-1 GP120，它似乎会导致小鼠的“临床前”脑疾病。 HIV-1 GP120被认为通过与巨噬细胞，星形胶质细胞和神经元上的表面受体相互作用而导致神经病发生。初步研究表明，携带天然GP120的HIV-1在培养中诱导小鼠星形胶质细胞中的炎症分子范围更大。我们假设ECOHIV所选的GP120段（例如V3）的表达将促进小鼠HIV-1介导的神经病发生。这里提出的探索性研究将以两个目的评估这一假设。在AIM 1中，我们将构建一系列嵌合Ecohiv，其中V3周围越来越大的HIV-1 GP120区域，包括负责与表面受体相互作用的嵌合，插入了ECOHIV的MLV ENV中。与ECOHIV相比，将对嵌合体进行体外和体内感染性以及它们在培养中诱导炎症蛋白诱导炎症蛋白的能力。我们构建的第一个这种嵌合体是Ecohiv-V3在体内具有感染性。在AIM 2中，我们将比较选定的GP120-表达嵌合体和EcoHIV，以诱导小鼠神经病发生。我们将确定体内病毒感染和神经浸润的动力学，大脑中HIV-1神经病发生的某些分子标记的表达以及神经病理学的发展。拟议的研究可能有助于定义HIV-1 GP120在小鼠中与缺乏GP120的病毒基因组的贡献的小鼠中HIV-1介导的神经病发生中的作用。该研究采用动物组织的动物模型，分子和病理分析以及分子病毒学和重组DMA技术。在较大的公共卫生环境中，我们的研究将在HIV-1发病机理研究中建立这种HIV-1感染动物模型的实用性，包括在中枢神经系统中，HIV-1疫苗的开发以及体内抗病毒化合物的便利性廉价测试。