REACTIVATION OF HERPES EYE DISEASE

疱疹性眼病复发

• 批准号: 2459188
• 负责人: DAVID J. FINK
• 金额: $ 15.68万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2459188
• 关键词: cornea disorder; gene expression; growth factor receptors; herpes simplex virus 1; host organism interaction; immunocytochemistry; in situ hybridization; latent virus infection; neurons; ocular herpes; phenotype; polymerase chain reaction; regulatory gene; relapse /recurrence

DESCRIPTION: Reactivation of herpes simplex virus (HSV) from latent infection of neurons in the trigeminal ganglion is a major cause of blindness. In order to define the molecular mechanisms of HSV reactivation from latency, we propose to study the hypothesis that the establishment of HSV latency reflects specific interactions between the phenotype of the individual neuron and promoter elements in the HSV genome, and the hypothesis that viral reactivation from latency results from identifiable alterations in the neuronal gene expression. Three specific aims are outlined. (1) To define the cellular phenotype of neurons containing latent viral genomes, comparing neurons in which the latent genomes express detectable LATs. The PI will use double- label immunocytochemistry, in situ hybridization and in situ PCR on serial 1 ym sections of ganglion to characterize cells into latent HSV genomes by neurotransmitter phenotype, expression of high affinity growth factor receptor, and the presence of transcriptional regulatory elements. (2) To define the mechanisms responsible for reactivation of HSV from latency on a cell specific basis. He will use similar methods after reactivating the virus containing the earliest reactivating genomes. (3) To empirically test the role of NGF and of the cellular immediate early genes c-fos and c-jun in reactivation he will construct a virus with the NGF gene driven by the ICP0 promoter to express NGF early in the reactivation cascade in order to test whether NGF expression blocks reactivation, and will construct a second recombinant lacking the cyclic AMP response element (CRE) in ICP0 promoter in order to test whether elevated expression of c-fos or c-jun by the cell activates ICP0 expression to cause entry of the latent genome into lytic cycle activity. The descriptive studies will define the phenotype of the cells harboring latent genomes and the alterations in that phenotype which occur early in the reactivation cascade. The viral constructs will empirically test the potential role of specific elements. Together, these studies will allow use to better understand the interaction between cellular and viral elements that results in reactivation of latent genomes from the trigeminal ganglion; an understanding that has important implications for strategies to prevent viral reactivation from latency, and the blindness which results from that reactivation.

描述：单纯疱疹病毒(HSV)从潜伏状态重新激活 三叉神经节神经元感染是导致 失明。为了明确单纯疱疹病毒的分子发病机制 从潜伏期重新激活，我们建议研究假设 HSV潜伏期的建立反映了 单纯疱疹病毒单个神经元和启动子元件的表型 基因组，以及潜伏期导致病毒重新激活的假设 神经基因表达的可识别变化。 概述了三个具体目标。(1)确定细胞表型 包含潜伏病毒基因组的神经元，比较 潜伏的基因组表达可检测到的LAT。PI将使用双倍- 标记免疫细胞化学、原位杂交和原位聚合酶链式反应 神经节连续1年切片将细胞定性为潜伏单纯疱疹病毒 基因组受神经递质表型影响，表达高亲和力生长 因子受体，以及转录调控元件的存在。 (2)明确HSV重新激活的机制 特定于单元的延迟。他将在之后使用类似的方法 重新激活包含最早重新激活基因组的病毒。 (3)经验性检验NGF和细胞即刻的作用 早期基因c-fos和c-jun的重新激活他将构建一种病毒 利用ICP0启动子驱动的NGF基因早期表达NGF 重新激活级联以测试NGF表达是否被阻断 重新激活，并将构建第二个缺少环状结构的重组 检测ICP0启动子中的AMP反应元件(CRE) 细胞c-fos或c-jun表达升高激活ICP0 导致潜伏基因组进入裂解周期的表达 活动。 描述性研究将确定细胞的表型 潜伏的基因组和早期发生的表型变化 在重新激活的级联中。病毒构建物将在实验中测试 特定元素的潜在作用。总而言之，这些研究将 允许用户更好地了解蜂窝和 导致潜伏基因组重新激活的病毒成分 三叉神经节；一个具有重要意义的理解 有关防止病毒因延迟而重新激活的策略，以及 失明是由这种重新激活造成的。