Immune cell dynamics during central nervous system viral infection

项目摘要/摘要 病毒可以在中枢神经系统中诱导多种疾病状态。一种特殊的疾病状态 我们小组的兴趣源于病毒诱发脑膜炎的能力。脑膜炎是一种可能致命的 由一长串人类病原体引起的疾病，通常与包括发烧在内的症状有关， 头痛，颈部僵硬，癫痫发作。目前，对病毒性肝炎患者几乎无能为力。 脑膜炎不是为了缓解症状。因此，我们建议对此有一个详细的了解 实时致病过程可以促进新干预措施的发展，以缓解症状和 防止永久性神经功能障碍/死亡。进行第一次致命的实时分析 脑膜炎，我们建议研究由淋巴细胞性脉络膜脑膜炎病毒引起的脑膜炎 (LCMV)-一种非细胞病变的小鼠，也是人类的病原体。小鼠脑内接种LCMV的实验研究 结果在6天内发生致命性脑膜炎，几乎完全由细胞毒性淋巴细胞(CTL)介导。 重要的是，这种疾病可以通过事先接种疫苗或免疫完全预防。的动态变化 感染控制失败或有效期间脑膜中的细胞相互作用尚未被研究。 之前。此外，在这个模型中，调节致命性损伤的确切机制并不完全 明白了。我们将利用最先进的技术组合，如病毒反向遗传学， 荧光标记的免疫细胞和双光子激光扫描显微镜结合原位 不同分子群染色观察LCMV感染小鼠的局部免疫细胞动态 大脑皮层和脑膜间隙。我们的假设是CTL对中枢神经系统星形胶质细胞网络的损害 在急性巨细胞病毒引起的脑膜炎期间导致全身性致死性发作，并且快速反应和 在免疫小鼠中通过激活记忆T细胞使用替代效应机制导致有限的 破坏星形胶质细胞网络，维持血脑屏障，并存活。这一假设将是 解决了三个集中的具体目标，涉及对CTL之间相互作用的第一次实时分析 和CNS靶点感染荧光标记的LCMV(Aim 1)，体内免疫突触的评估 CNS CTL靶向/损伤的形成和分子机制(目标2)，以及实时 确定在中枢神经系统内工作的记忆T细胞提供的保护(目标3)。

Project Summary/Abstract Viruses can induce a variety of disease states in the central nervous system. A disease state of particular interest to our group stems from the ability of viruses to induce meningitis. Meningitis is a potentially fatal disorder induced by a long list of human pathogens 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 therefore 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 / fatalities. To conduct the first real time analyses of fatal meningitis, 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 techniques such as 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 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 in three focused specific aims that involve the first real time analyses of interactions between CTL and CNS targets infected by fluorescently-tagged LCMV (aim 1), in vivo evaluation of immunological synapse formation and molecular mechanisms involved in CNS CTL targeting / damage (aim 2), and real time determination of the protection afforded by memory T cells operating within the CNS (aim 3).