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)，在神经节潜伏 神经轴的所有水平的神经元，几十年后重新激活，引起带状疱疹(带状疱疹)和 带状疱疹后遗神经痛(PHN)，尤其是老年人。尽管有带状疱疹疫苗可用，即使 美国60岁的每个成年人都接种了疫苗，每年仍有50万带状疱疹病例， 其中约20万人将经历VZV引起的PHN、中风、失明或脊髓病 重新激活。 疫苗接种的另一种选择是基于对病毒重新激活的深入了解 潜伏期病毒核酸和基因表达的物理状态及其变化 重新激活。我们和其他人已经证明了VZV IE63，由OPEN编码的即刻早期(IE)蛋白 读取帧63是VZV潜伏期的一致标志。基于这样一个事实，IE63，一个主要的 在生殖性感染过程中，核磷蛋白仅定位于潜伏期- 在其磷酸化状态未知的情况下，我们假设IE63的易位 从细胞质到细胞核导致IE63功能改变和病毒的启动 重新激活。我们将通过鉴定IE63的细胞质和核形式以及它们的 IE63在人神经细胞和非神经细胞培养中的功能定位(Aim1) 病毒再激活时的神经节(AIM2)和VZV基因转录模式及其表观遗传学 移植的人三叉神经节(Aim3)再激活过程中的调节。这些研究将提供 VZV在人类体内重新激活的第一张全面图片。 我们是唯一有资格进行这些研究的人，因为我们有源源不断的人力供应 在尸检中获得的神经节显示，90%以上的人类神经节是潜伏感染的 对于VZV，VZV在移植后重新激活，并且优化的GeXP技术检测到 低丰度VZV基因转录本。此外，我们还构建了重组鼠抗IE63抗体 识别IE63核或细胞质形式的抗体。 我们对人类TG中VZV重新激活机制的深入分析将确定新的靶点 防止病毒重新激活的治疗干预。

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.