Immune cell dynamics during central nervous system viral infection

中枢神经系统病毒感染期间的免疫细胞动力学

• 批准号: 7581118
• 负责人: Juan C. de la Torre
• 金额: $ 48.19万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-05 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7581118
• 关键词: Acute; Address; Antibodies; Antigen-Presenting Cells; Arts; Astrocytes; Blood - brain barrier anatomy; Brain; Brain region; CD8B1 gene; CXCL10 gene; CXCR3 gene; CXCR6 gene; Calcium Oscillations; Calcium Signaling; Cell Communication; Cells; Central Nervous System Viral Diseases; Cerebral cortex; Containment; Cytotoxic T-Lymphocytes; Development; Disease; Epilepsy; Evaluation; Fever; Fostering; Goals; Headache; Human; Human Virus; ITGAX gene; Image; Immune; Immunization; Immunosuppressive Agents; In Situ; Infection; Infection Control; Inflammatory; Injury; Integrins; Intercellular Adhesion Molecule 2; Intercellular adhesion molecule 1; Intervention; Label; Laboratories; Laser Scanning Microscopy; Lymphocyte; Lymphocytic choriomeningitis virus; Maintenance; Mediating; Meningeal; Meninges; Meningitis; Methods; Microglia; Modeling; Molecular; Mus; Neck; Neuraxis; Neurologic; Neurologic Dysfunctions; Pathogenesis; Patients; Principal Investigator; Process; Proteins; Reaction; Regulation; Role; Route; Seizures; Staining method; Stains; Surface; Symptoms; System; T memory cell; T-Lymphocyte; Time; Upper arm; Vaccinated; Vaccination; Vascular Cell Adhesion Molecule-1; Viral; Viral meningitis; Virus; Virus Diseases; cell killing; chemokine; chemokine receptor; cranium; cytotoxic; disability; granzyme B; immunological synapse; immunological synapse formation; in vivo; injured; migration; novel; pathogen; perforin; positional cloning; prevent; programs; purge; response; time use; two-photon

Program Director/Principal Investigator (Last, First, Middle): De La Torre, Juan C. 1R01 AI075298-01A2 Viruses can induce a variety of disease states in the central nervous system. Meningitis is a potentially fatal disorder induced by a long list of human pathogens, including viruses, and is often associated with symptoms that include fever, headache, stiffness of the neck, and seizures. Presently, very little can be done for patients with viral meningitis other than to relieve symptoms. We propose that a detailed understanding of this pathogenic process in real time may foster the development of novel interventions to alleviate symptoms and prevent permanent neurological dysfunction and fatalities. To accomplish this goal we propose to study the well-described meningitis induced by lymphocytic choriomeningitis virus (LCMV), a noncytopathic mouse as well as human pathogen. Intracerebral inoculation of mice with LCMV results in a fatal meningitis within 6 days that is mediated almost entirely by cytotoxic lymphocytes (CTL). Importantly, this disease can be completely prevented by prior vaccination or immunization. The dynamics of cellular interactions in the meninges during failed or effective control of infection have not been studied previously. Moreover, the precise mechanisms that mediate fatal injury in this model are not entirely understood. We will utilize a combination of state-of-the art viral reverse genetics, fluorescently-tagged immune cells, and two-photon laser scanning microscopy in combination with in situ staining for different molecular species to follow the local immune cell dynamics in the LCMV-infected mouse cerebral cortex and meningeal space. Our hypothesis is that CTL damage to astrocyte networks in the CNS leads to generalized fatal seizure during acute LCMV-induced meningitis, and that rapid responsiveness and the use of alternative effector mechanisms by activated memory T cells in vaccinated mice results in limited damage to astrocyte networks, maintenance of the blood brain barrier, and survival. This hypothesis will be addressed by the two focused following specific aims: 1) Completion of the first real time analyses of interactions between CTL and CNS targets infected by fluorescently-tagged LCMV, and 2) in vivo evaluation of immunological synapse formation and molecular mechanisms involved in CNS CTL targeting and damage.

项目总监/首席研究员（最后、第一、中间）：De La Torre, Juan C. 1R01 AI075298-01A2 病毒可以诱发中枢神经系统的多种疾病状态。脑膜炎是一种潜在致命性疾病，由包括病毒在内的多种人类病原体引起，通常伴有发烧、头痛、颈部僵硬和癫痫发作等症状。目前，除了缓解症状之外，对病毒性脑膜炎患者几乎无能为力。我们建议，实时详细了解这种致病过程可能会促进新干预措施的开发，以减轻症状并预防永久性神经功能障碍和死亡。为了实现这一目标，我们建议研究由淋巴细胞性脉络膜脑膜炎病毒（LCMV）（一种非细胞病变的小鼠以及人类病原体）引起的脑膜炎。小鼠脑内接种 LCMV 会在 6 天内导致几乎完全由细胞毒性淋巴细胞 (CTL) 介导的致命性脑膜炎。重要的是，这种疾病可以通过事先接种疫苗或免疫接种来完全预防。先前尚未研究过感染控制失败或有效期间脑膜中细胞相互作用的动态。此外，在该模型中介导致命伤害的精确机制尚不完全清楚。我们将利用最先进的病毒反向遗传学、荧光标记免疫细胞和双光子激光扫描显微镜的组合，并结合不同分子物种的原位染色，以跟踪 LCMV 中的局部免疫细胞动态。感染小鼠大脑皮层和脑膜腔。我们的假设是，在急性 LCMV 诱导的脑膜炎期间，CTL 对中枢神经系统星形胶质细胞网络的损伤导致全身性致命性癫痫发作，而免疫小鼠中激活的记忆 T 细胞的快速反应和使用替代效应机制会导致对星形胶质细胞网络的有限损伤、维持血脑屏障、维持生存。这一假设将通过以下两个具体目标来解决：1）完成对荧光标记的 LCMV 感染的 CTL 和 CNS 靶标之间相互作用的首次实时分析，以及 2）对免疫突触形成和相关分子机制的体内评估CNS CTL 靶向和损伤。