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/首席调查员（最后，第一，中间）：De la Torre，Juan C. 1R01 AI075298-01A2 病毒可以在中枢神经系统中诱导各种疾病状态。脑膜炎是一长串人类病原体（包括病毒）引起的潜在致命疾病，通常与包括发烧，头痛，颈部僵硬和癫痫发作有关的症状有关。目前，除了缓解症状外，其他病毒脑膜炎患者几乎无法做到。我们建议实时对这种致病过程的详细理解可以促进新的干预措施的发展，以减轻症状并防止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绒毛膜炎病毒（LCMV），一种非肿瘤性小鼠以及人类病原体诱导的良好描述的脑膜炎。脑内接种LCMV的小鼠在6天内导致致命性脑膜炎，几乎完全由细胞毒性淋巴细胞（CTL）介导。重要的是，可以通过事先疫苗接种或免疫完全预防该疾病。之前尚未研究脑膜相互作用在失败或有效控制感染过程中的动力学。此外，在该模型中介导致命伤害的确切机制尚不完全理解。我们将利用最先进的病毒反向遗传学，荧光标记的免疫细胞和两光子激光扫描显微镜结合使用，以及不同分子物种的原位染色，以遵循LCMV感染的小鼠小鼠小鼠CEREBRAL CEREBRAL CORTEX和脑膜层次的局部免疫细胞动力学。我们的假设是，CNS中的CTL对星形胶质细胞网络的损害导致急性LCMV诱发的脑膜炎期间的全身致命癫痫发作，并且疫苗接种的记忆T细胞的快速反应性以及使用替代效应机制的使用，导致疫苗接种的小鼠中的替代效应机制导致对体形胶质细胞网络的有限损害，脑脑脑障碍和存活率有限。该假设将通过两个重点以特定目的来解决：1）CTL和CNS靶标之间相互作用的第一个实时分析，这些靶标感染了荧光标记的LCMV，而2）在体内评估免疫突触形成和CNS CNS CNS CNS CNS CNS CNS CNS涉及的分子机制。