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，它们的成功是由于它们的能力 建立潜伏感染，并调节宿主的免疫力。这些能力让病毒 化疗剂难以治愈，免疫系统难以清除。虽然延迟和 对疱疹病毒的免疫已经被深入研究，控制建立的机制， 潜伏期的维持和再激活仍然不清楚，特别是对于单纯疱疹病毒（HSV）。这是由于， 在某种程度上，由于体内模型（主要是小鼠）的高度复杂性，虽然信息量很大，但无法提供信息， 准确地说，HSV和神经元之间的独特关系，它的潜伏期的目标细胞。最近我们 对潜伏期的理解已经通过发展生理相关的 使用原代培养的神经元的体外系统， 系统来询问潜伏和免疫的分子基础。项目3的过度使用方法是 使用和开发相关的体外和体内模型系统，以证明内在的，先天的， 适应性免疫和决定潜伏/再激活轴平衡的病毒对策。目的 1，项目1和2，检查自噬体簇的结构和功能（内在免疫） 在潜伏期的建立和维持期间在感觉神经元中诱导。我们将探索这些 用高分辨率显微镜观察新发现的结构，并表征基因表达和染色质 HSV在簇阳性神经元中的模式。目的2研究病毒调节活性氧的作用 种和线粒体运动的能力，HSV的建立，并重新激活从培养的神经元 潜伏期和干扰素依赖性先天免疫应答的发生。在项目1和2中， 本研究的目的还将研究HSV蛋白ICP0和ICP34.5在调节这些反应中的作用， 它们对基因表达和染色质模式的影响。目的3解决了抗体是 HSV-1复制、发病机制和神经系统潜伏期的关键免疫决定因素。的 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， 和再活化，包括对基因表达和染色质的影响。项目1， 实验性疫苗和抗体不仅能保护小鼠免于HSV感染后死亡， 挑战，而且还来自与新生儿感染相关的长期行为病理学， 评估。因此，一般的做法是继续与项目1和项目2合作，并利用核心B， 开发和利用我们的初级神经元和新生儿感染模型系统， 核心A的基因表达和染色质模式，以及实验免疫。这些办法将 确定免疫的不同分支如何控制潜伏期，以及HSV如何对抗这些反应。这 该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.