ROLE OF VZV IE63 IN VZV LATENCY AND REACTIVATION

水痘带状疱疹病毒（VZV）引起儿童水痘（水痘），在神经节细胞中潜伏， 神经轴各级的神经元，几十年后重新激活，引起带状疱疹， 带状疱疹后神经痛（PHN），尤其是老年人。虽然带状疱疹疫苗是可用的，即使 即使美国60岁以上的成年人都接种了疫苗，每年仍有约50万例带状疱疹病例， 其中约20万人将经历由VZV引起的PHN、中风、失明或脊髓病 重新激活 疫苗接种的一种替代方法是在深入了解 潜伏期病毒核酸和基因表达的物理状态及其在 重新激活我们和其他人已经表明，VZV IE63，即由开放式编码的立即早期（IE）蛋白， 阅读帧63是VZV延迟的一致标志。基于这样一个事实，即IE63，一个主要的 核磷蛋白在生产性感染，只位于细胞质中的潜伏- 感染的神经元，其磷酸化状态是未知的，我们假设IE63易位， 从细胞质到细胞核的转移导致IE63功能的改变和病毒的启动 重新激活我们将通过鉴定IE63的细胞质和细胞核形式以及它们的表达来检验这一假设。 在培养的神经元和非神经元细胞中的功能（Aim 1），IE63在人类中的细胞定位 VZV基因的转录模式及其表观遗传 调节过程中重新激活的人三叉神经节（Aim 3）。这些研究将提供 这是第一张关于VZV在人体内重新激活的全面图片。 我们是唯一有资格进行这些研究，因为我们有一个持续的供应， 在尸检中获得的神经节，已经表明来自超过90%的人类的神经节是潜伏感染的， 使用VZV时，VZV重新激活发生在重新激活之后，并且优化的GeXP技术检测到 低丰度VZV基因转录物。此外，我们构建了重组小鼠抗IE63抗体， 识别IE63的细胞核或细胞质形式的抗体。 我们对VZV在人TG中再激活机制的深入分析将为VZV的治疗确定新的靶点。 治疗干预以防止病毒再活化。

Varicella zoster virus (VZV) causes childhood varicella (chickenpox), becomes latent in ganglionic neurons at all levels of the neuraxis and reactivates decades later to cause zoster (shingles) and postherpetic neuralgia (PHN), especially in the elderly. Although a zoster vaccine is available, even if every adult in the U.S. >60 years were vaccinated, there would still be -500,000 zoster cases annually, about 200,000 of whom will experience PHN, stroke, blindness or myelopathy caused by VZV reactivation. An alternative to vaccination is prevention of virus reactivation based on an in-depth understanding of the physical state of viral nucleic acid and gene expression during latency and their changes during reactivation. We and others have shown that VZV IE63, the immediate early (IE)protein encoded by open reading frame 63, is a consistent hallmark of VZV latency. Based on the fact that IE63, a predominantly nuclear phosphoprotein during productive infection, is located exclusively in the cytoplasm of latently- infected neurons where its phosphorylation state is unknown, we hypothesize that translocation of IE63 from the cytoplasm to the nucleus results in altered IE63 function and initiation of virus reactivation. We will test this hypothesis by identifying cytoplasmic and nuclear forms of IE63 and their functions in neuronal and non-neuronal cells in culture (Aim1), the cellular location of IE63 in human ganglia during virus reactivation (Aim2), and the pattern of VZV gene transcription and their epigenetic regulation during reactivation in explanted human trigeminal ganglia (Aim3). These studies will provide the first comprehensive picture of VZV reactivation in humans. We are uniquely qualified to carry out these studies since we have a continuous supply of human ganglia obtained at autopsy, have shown that ganglia from more than 90% of humans are latently infected with VZV,that VZV reactivation occurs after explantation, and that optimized GeXP technology detects low-abundance VZV gene transcripts. Further, we have constructed recombinant mouse anti-IE63 antibodies that recognize nuclear or cytoplasmic forms of IE63. Our in-depth analysis of the mechanism of VZV reactivation in human TG will identify new targets for therapeutic intervention to prevent virus reactivation.