Project 3 - Innate immunity and the HSV lytic/latent balance

疱疹病毒是人类发病率和死亡率的主要来源，它们的成功归功于它们的能力 以确定潜在感染，并调节其宿主的免疫力。这些能力使病毒 化疗药物难以治愈，免疫系统难以清除。虽然延迟和 对疱疹病毒的免疫进行了深入的研究，控制建立的机制， 潜伏期的维持和重新激活仍然不清楚，特别是对于单纯疱疹病毒(HSV)。这是应该的， 在一定程度上，这是由于体内(主要是小鼠)模型的高度复杂性，这些模型虽然提供了高度的信息，但未能提供 关于HSV和神经元之间的独特关系，神经元是HSV潜伏期的靶细胞。最近，我们的 对潜伏期的理解由于生理上相关的发展而显著增强 使用原代培养神经元的体外系统，可以与更传统的体内更多的使用 系统询问潜伏期和免疫力的分子基础。项目3的总体方法是 使用和开发相关的体外和体内模型系统，以展示内在的、天生的和 决定潜伏期/再激活轴平衡的适应性免疫和病毒对策。目标 1，项目1和2，检查自噬体簇的结构和功能(固有免疫) 在潜伏期的建立和维持期间在感觉神经元中诱导的。我们将对这些进行调查 用高分辨率显微镜新发现的结构，并表征基因表达和染色质 簇状阳性神经元中单纯疱疹病毒感染的类型。目的2研究病毒对活性氧的调节作用 培养神经元中的种类和线粒体运动对HSV建立和重新激活能力的影响 潜伏期，以及干扰素依赖的先天免疫反应的起源。对于项目1和2，这一点 AIM还将研究HSV蛋白ICP0和ICP34.5在调节这些反应中的作用，并 它们对基因表达和染色质模式的影响。目标3提出的假设是抗体是 神经系统中HSV-1复制、发病机制和潜伏期的关键免疫决定因素。这个 将定义感觉神经节内抗体的活性，以及它们在控制复制、潜伏期、 重新激活，包括对基因表达和染色质的影响，将被确定。在项目1中， 实验性疫苗和抗体不仅能保护小鼠免受单纯疱疹病毒感染后的死亡 挑战，也来自于与新生儿感染相关的长寿行为病理 评估过了。然后，一般方法是继续与项目1和2合作，并使用核心B，以 开发和利用我们的初级神经元和新生儿感染模型系统，与分析接口 基因表达和染色质模式的核心A，以及实验免疫。这些方法将 确定免疫的不同分支如何控制潜伏期，以及HSV如何应对这些反应。这 框架将确定新的途径，以帮助干预这种持久性人类病原体的生命周期。

Herpesviruses are a major source of morbidity and mortality in humans and their success is due to their ability to establish latent infections, and to modulate the immunity of their hosts. These abilities render the virus refractory to cure by chemotherapeutic agents, and to clearance by the immune system. While latency and immunity to herpesviruses have been intensely studied, the mechanisms that control establishment, maintenance and reactivation of latency remain obscure, especially for herpes simplex virus (HSV). This is due, in part, to the high complexities of in vivo (mostly mouse) models that while highly informative, fail to inform precisely about the unique relationship between HSV and the neuron, its target cell for latency. Recently, our understanding of latency has been significantly enhanced by the development of physiologically relevant in vitro systems employing primary cultured neurons that can be used in combination with more traditional in vivo systems to interrogate the molecular basis of latency and immunity. The over-arching approach of Project 3 is to use and develop relevant in vitro and in vivo models systems to demonstrate roles for intrinsic, innate, and adaptive immunity and viral countermeasures that determine the balance of the latency/reactivation axis. Aim 1, with Projects 1 and 2, examines the structure and function of autophagosomes clusters (intrinsic immunity) that are induced in sensory neurons during the establishment and maintenance of latency. We will probe these newly-discovered structures with high resolution microscopy, and characterize gene expression and chromatin patterns of HSV in cluster-positive neurons. Aim 2 examines the role of viral modulation of reactive oxygen species and mitochondrial movement in cultured neurons on the ability of HSV to establish, and reactivate from latency, and on the genesis of the interferon-dependent innate immune response. With Projects 1 and 2, this aim will also examine the roles of HSV proteins ICP0 and ICP34.5 in the modulation of these responses and their effects on gene expression and chromatin patterns. Aim 3 addresses the hypothesis that antibodies are key immune determinants of HSV-1 replication, pathogenesis, and latency in the nervous system. The activities of antibodies within sensory ganglia will be defined, and their roles in the control of replication, latency, and reactivation, including effects on gene expression and chromatin, will be determined. With Project 1, the ability of an experimental vaccine and antibodies to protect mice not only from mortality following HSV challenge, but also from the long-lived behavioral pathology associated with neonatal infection will be assessed. The general approach then is to continue collaborations with Projects 1 and 2, and, using Core B, to develop and exploit our primary neuron and neonatal infection model systems interfaced with analyses with Core A of gene expression and chromatin patterns, and experimental immunization. These approaches will determine how different branches of immunity control latency, and how HSV counters these responses. This framework will identify novel pathways to help intervene in the lifecycle of this persistent human pathogen.