Varicella Zoster Virus Latency in Human Ganglia

水痘带状疱疹病毒在人类神经节中的潜伏期

• 批准号: 6746034
• 负责人: RANDALL J. COHRS
• 金额: $ 33.12万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-12-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6746034
• 关键词: antibody; ganglion cell; ganglions; gene expression; genetic mapping; genetic transcription; human subject; immunocytochemistry; latent virus infection; molecular cloning; nervous system infection; neurotropic virus; nucleic acid sequence; patient oriented research; protein localization; protein protein interaction; recombinant proteins; tissue /cell culture; transfection; varicella zoster virus; virus genetics; virus infection mechanism; virus protein; virus replication; yeast two hybrid system

Varicella zoster virus (VZV) is an ubiquitous human pathogen. Primary infection causes childhood chickenpox, leading to latency in cranial nerve, dorsal root and autonomic ganglia. VZV reactivation, frequent in the elderly and immunocompromised population, results in significant neurological disease. Zoster and postherpetic neuralgia predominate, but many humans develop myelitis, segmental motor weakness, cranial nerve palsies, and a severe, often fatal vasculopathy in the brain. We have also shown that VZV reactivation can produce chronic intense pain in the absence of rash. Although the mechanism controlling VZV latency is not understood, it is likely to depend upon virus gene transcription, thus providing the rationale for our hypothesis that VZV gene expression in human ganglia during latency functions to maintain latent infection. Identification of the VZV genes expressed during latency is critical to understanding latent infection. Until the advent of high throughput array-based technology, such analyses were not feasible, and <20% of the VZV genome has been analyzed for latent virus transcripts. Further, merely identifying the latently transcribed VZV genes is insufficient. Since control of VZV latency is likely to involve virus proteins, characterization of latently expressed viral proteins is also critical to understanding virus gene regulation. Thus, our long-term goal is the detailed characterization, including functional analysis, of VZV genes expressed in latently infected human ganglia. Our specific aims will: (1) identify latently transcribed VZV genes by transcriptional array analysis, confirm the authenticity of the transcripts by sequencing, and determine the abundance of the virus transcripts by fluorescence-based quantitative (real-time) RT-PCR; (2) construct high affinity epitope-tagged recombinant antibodies to colocalize latently expressed VZV proteins by in situ immunohistochemistry in sections of human ganglia; and (3) initiate protein function analysis using 2-hybrid systems to identify and map protein-protein interactions. We will begin with VZV IE63, the most prevalent and abundant virus transcript detected to date during latency. An in-depth understanding of VZV gene expression and function in latently infected human ganglia will lead to testable models of latent virus gene regulation (in the developing simian varicella virus model) and to therapies designed to reduce morbidity and mortality associated with reactivation of this highly neurotropic human pathogen.

水痘带状疱疹病毒（VZV）是一种普遍存在的人类病原体。原发感染引起儿童水痘，导致脑神经、背根和自主神经节潜伏。 VZV 重新激活常见于老年人和免疫功能低下人群，会导致严重的神经系统疾病。带状疱疹和带状疱疹后神经痛占主导地位，但许多人会出现脊髓炎、节段性运动无力、颅神经麻痹以及严重的、经常发生的神经痛。 致命的大脑血管病变。我们还表明，VZV 重新激活可以在没有皮疹的情况下产生慢性剧烈疼痛。尽管控制VZV潜伏期的机制尚不清楚，但它很可能依赖于病毒基因转录，从而为我们的假设提供了理论依据，即潜伏期人类神经节中VZV基因的表达发挥着维持潜伏感染的作用。鉴定潜伏期间表达的 VZV 基因对于了解潜伏感染至关重要。在基于高通量阵列的技术出现之前，此类分析是不可行的，并且已分析了 <20% 的 VZV 基因组的潜在病毒转录本。此外，仅鉴定潜在转录的 VZV 基因是不够的。由于 VZV 潜伏期的控制可能涉及病毒蛋白，因此潜伏表达病毒蛋白的表征对于了解病毒基因调控也至关重要。因此，我们的长期目标是对表达的 VZV 基因进行详细表征，包括功能分析 在潜伏感染的人类神经节中。我们的具体目标是：（1）通过转录阵列分析识别潜在转录的VZV基因，通过测序确认转录本的真实性，并通过基于荧光的定量（实时）RT-PCR确定病毒转录本的丰度； (2)构建高亲和力表位标记的重组抗体，通过原位免疫组化在人神经节切片中共定位潜在表达的VZV蛋白； (3) 使用 2-杂交系统启动蛋白质功能分析，以识别和绘制蛋白质-蛋白质相互作用图谱。我们将从 VZV IE63 开始，它是迄今为止在潜伏期间检测到的最普遍、最丰富的病毒转录本。深入了解潜伏感染的人类神经节中的 VZV 基因表达和功能，将产生潜伏病毒基因调控的可测试模型（在正在开发的猿水痘病毒模型中），以及旨在降低与这种高度病毒重新激活相关的发病率和死亡率的治疗方法。嗜神经人类病原体。