NEUROTROPISM AND MICROGLIAL INVASION

向神经性和小胶质细胞侵袭

• 批准号: 6481248
• 负责人: Francisco Gonzalez-Scarano
• 金额: $ 10.35万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6481248
• 关键词: AIDS dementia complex; HIV envelope protein gp120; HIV infections; antiAIDS agent; apoptosis; calcium; chemokine; human immunodeficiency virus 1; human tissue; intracellular transport; macrophage; microglia; molecular cloning; monocyte; natural gene amplification; neuropathology; neurotropic virus; neutralizing antibody; polymerase chain reaction; tissue /cell culture; virulence; virus cytopathogenic effect; virus genetics; virus load; virus replication

Microglia are critical to the primary complications of the human immunodeficiency virus (HIV-1) in the central nervous system (CNS), since they are the most commonly infected cell and their infection represents the majority of the viral load in the CNS. This proposal will center on the biology of HIV-1 in microglial in order to develop a better understanding of the role of the virus in the development of this complication, and to identify potential, CNS-specific, treatment strategies in collaboration with the other components of this program. In the first specific aim we will continue our studies on microglial- tropism of HIV-1 isolates using primary isolates from adults and children, and an isolate adapted to microglia bu sequential passage. We will first use a PCR-based assay to analyze the sequential steps of HIV-1 infection in microglia. For those isolates (like the microglia-adapted HIV-1/BORI- 15) which demonstrate a rapid entry phenotype, we will molecularly clones the envelopes, and define the mechanism of enhanced cellular penetration using molecular and biochemical (binding) assays. Where tropism for microglial cells is related to post-entry steps, other portions of the provirus, or the entire proviral genome, will be cloned. We will then determine whether isolates that do not replace to high levels in microglia can nevertheless establish a chronic infection. These isolates will then be used in a SCID-hu model in another Project. In the second specific aim we will determine whether the envelope proteins, and specifically gp120 from isolates with HIV encephalopathy can mediate changes in intracellular free Ca2+ concentrations in monocyte-derived macrophages (MDM), microglia, and other neural cells. Those gp120s that induce intracellular signals will be tested for their ability to mediate apoptosis. In the third specific aim we will determine whether microglial infection by certain HIV isolates results in increased production of chemokines, which could be responsible for increased cellular trafficking into the CNS, and potential amplification of a chronic infection. The result from these experiments will strengthen knowledge about the interactions between HIV-1 and microglial cells.

小胶质细胞对于中枢神经系统（CNS）中人类免疫缺陷病毒（HIV-1）的主要并发症至关重要，因为它们是最常见的细胞，其感染代表了CNS中大部分病毒载量。该提案将集中在小胶质细胞中HIV-1的生物学上，以便更好地了解病毒在这种并发症发展中的作用，并与该计划的其他组成部分合作确定潜在的，CNS特异性的治疗策略。在第一个具体目的中，我们将使用成人和儿童的原代分离株以及适合小胶质细胞BU顺序通过的分离株来继续研究HIV-1分离株的小胶质细胞 - 对流。我们将首先使用基于PCR的测定法分析小胶质细胞中HIV-1感染的顺序步骤。对于这些分离株（如适应小胶质细胞的HIV-1/BORI-15），该分离株证明了快速进入表型，我们将分子克隆信封，并使用分子和生化（结合）测定法定义增强细胞穿透的机制。如果小胶质细胞的热态与进入后步骤有关，则将克隆前病毒或整个病毒基因组的其他部分或整个病毒基因组。然后，我们将确定未取代小胶质细胞中高水平的分离株是否可以建立慢性感染。然后，这些分离物将在另一个项目的SCID-HU模型中使用。在第二个特定目的中，我们将确定包膜蛋白，特别是来自患有HIV脑病的分离株的GP120是否可以介导单核细胞衍生的巨噬细胞（MDM），小胶质细胞和其他神经细胞的细胞内游离Ca2+浓度的变化。那些诱导细胞内信号的GP120将测试其介导凋亡的能力。在第三个特定目的中，我们将确定某些HIV分离株的小胶质细胞感染是否会导致趋化因子的产生增加，这可能导致细胞运输增加到中枢神经系统中，并潜在地扩增慢性感染。这些实验的结果将加强有关HIV-1与小胶质细胞之间相互作用的知识。