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。原发性感染会导致儿童水痘，导致颅神经，背根和自主神经节的潜伏期。 VZV重新激活，在老年人和免疫功能低下的人群中频繁导致明显的神经系统疾病。带状疱疹和骨后神经痛占主导地位，但许多人会出现髓炎，节段运动无力，颅神经麻痹以及严重的，通常 大脑中的致命血管病。我们还表明，在没有皮疹的情况下，VZV重新激活会产生慢性剧烈疼痛。尽管无法理解控制VZV潜伏期的机制，但它很可能取决于病毒基因转录，因此为我们的假设提供了理由，即在潜伏期起作用期间，人神经神经节中VZV基因表达以维持潜在感染。在潜伏期中表达的VZV基因的鉴定对于理解潜在感染至关重要。在基于高吞吐量阵列的技术的出现之前，这种分析是不可行的，并且已经分析了<20％的VZV基因组的潜在病毒转录本。此外，仅识别延伸的vzv基因是不足的。由于对VZV潜伏期的控制可能涉及病毒蛋白，因此对潜在表达的病毒蛋白的表征对于理解病毒基因调节也是至关重要的。因此，我们的长期目标是表达的VZV基因的详细表征，包括功能分析 在潜在感染的人神经节中。我们的具体目的将：（1）通过转录阵列分析确定延伸的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确定病毒转录物的丰度； （2）构建高亲和力标记的重组抗体，通过原位免疫组织化学在人神经节的部分中通过原位免疫组织化学共定位。 （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。我们将从VZV IE63开始，这是迄今为止在潜伏期期间检测到的最普遍，最丰富的病毒转录本。对潜在感染的人神经节中VZV基因表达和功能的深入了解将导致可测试的潜在病毒基因调节模型（在发展中的猿猴Varicella病毒模型中）以及旨在降低与这种高度神经性人类人类病原体反应活力相关的发病率和死亡率的疗法。