Role of astrocyte infection in viral neurovirulence

星形胶质细胞感染在病毒神经毒力中的作用

• 批准号: 10011753
• 负责人: VINCENT R RACANIELLO
• 金额: $ 20.25万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-10 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10011753
• 关键词: Acute; Address; Alphavirus; Amino Acid Sequence; Antibodies; Astrocytes; Brain; Brain Injuries; Cells; Cessation of life; Complex; Development; Disease; Disease Outcome; Enterovirus; Equilibrium; Exposure to; Flavivirus; Glial Fibrillary Acidic Protein; Goals; Health; Human poliovirus; Immune; Indirect Immunofluorescence; Infection; Inflammatory; Invaded; Knowledge; Lansing Virus; Lead; Lipopolysaccharides; Microglia; Molecular; Morphology; Motor Neurons; Mus; Neuraxis; Neuroglia; Neurons; Neurotoxins; Neurotropism; Oligodendroglia; Paralysed; Pathogenesis; Phenotype; Plaque Assay; Poliomyelitis; RNA Viruses; Recombinants; Role; Route; Serotyping; Severities; Site; Slice; Spinal Cord; Stress; System; Tissues; Tropism; Viral; Virus; Virus Diseases; Virus Replication; West Nile virus; Wild Type Mouse; Work; astrogliosis; cell type; chikungunya; cytokine; design; experimental study; insight; mouse model; nervous system disorder; neuron loss; neuropathology; neurotoxic; neurotropic virus; neurovirulence; poliovirus receptor; recombinant virus; response; spinal cord and brain injury; stem

Project summary Understanding how infection with the mouse-adapted Lansing isolate of poliovirus type 2 (PV-P2/L) infects wild-type, nontransgenic mice and lead to poliomyelitis has remained an unanswered question for more than 50 years. Remarkably, the ability of this poliovirus strain to infect wild-type mice can be conferred to other virus isolates by the exchange of a 10 amino acid sequence from the VP1 B-C loop of PV-P2/L. Intracerebral infection of wild-type mice with this recombinant virus, PV-414, also leads to the development of paralysis Similarly to PV-P2/L, virus replication of PV-414 is limited to the brain and spinal cord. Neurologic disease associated with virus infection was thought to be due to direct virus killing of motor neurons. To our surprise, we found that these viruses replicate not in neurons but in astrocytes of the brain. In this application, we hypothesize that astrocytes, a major class of central nervous system glia, when virally infected undergo reactive astrogliosis, differentiating to A1 neurotoxic astrocytes. Stimulation of A1 astrocytes is pro- inflammatory and leads to secretion of numerous cytokines that contribute to neuronal and oligodendrocyte death, facilitating development of many neuropathologies. We propose to identify the site of replication within the central nervous system of PV-P2/L and PV-414 using organotypic brain and spinal cord slice cultures derived from wild-type mice. To determine if astrocytes are also sites of PV-414 infection within the spinal cord, organotypic cultures will generated from multiple regions of the spinal cord and infected with PV-414. Indirect immunofluorescence using antibodies against specific cell types will be used to characterize the infected cell. Identification of the soluble neurotoxin(s) secreted by neurotoxic astrocytes during poliovirus infection, and the mechanism by which neuronal death occurs during this response, will provide additional insight into the mechanism of virus induced tissue damage, a more complete understanding of poliovirus pathogenesis, and define the function of reactive astrocytes during viral infection.

项目概要 了解 2 型脊髓灰质炎病毒兰辛分离株 (PV-P2/L) 是如何感染小鼠的 野生型、非转基因小鼠和导致脊髓灰质炎的问题多年来一直是一个悬而未决的问题 50 年。值得注意的是，这种脊髓灰质炎病毒株感染野生型小鼠的能力可以赋予其他病毒 通过从 PV-P2/L 的 VP1 B-C 环交换 10 个氨基酸序列来分离。脑内 野生型小鼠感染这种重组病毒 PV-414 也会导致瘫痪 与PV-P2/L类似，PV-414的病毒复制仅限于大脑和脊髓。神经系统疾病 与病毒感染相关的现象被认为是由于病毒直接杀死运动神经元。令我们惊讶的是， 我们发现这些病毒不是在神经元中复制，而是在大脑的星形胶质细胞中复制。在这个应用程序中，我们 假设星形胶质细胞（中枢神经系统胶质细胞的一类主要类型）在受到病毒感染时会经历 反应性星形胶质细胞增生，分化为 A1 神经毒性星形胶质细胞。 A1 星形胶质细胞的刺激是亲 炎症并导致大量细胞因子的分泌，这些细胞因子有助于神经元和少突胶质细胞 死亡，促进许多神经病理学的发展。我们建议确定内的复制位点 使用器官型脑和脊髓切片培养的 PV-P2/L 和 PV-414 的中枢神经系统 源自野生型小鼠。为了确定星形胶质细胞是否也是脊髓内 PV-414 感染的部位， 器官型培养物将从脊髓的多个区域产生并感染 PV-414。间接 使用针对特定细胞类型的抗体的免疫荧光将用于表征受感染的细胞。 脊髓灰质炎病毒感染期间神经毒性星形胶质细胞分泌的可溶性神经毒素的鉴定，以及 在此反应期间神经元死亡发生的机制将提供更多关于 病毒引起的组织损伤的机制，对脊髓灰质炎病毒发病机制有更完整的了解，以及 定义病毒感染期间反应性星形胶质细胞的功能。