REACTIVATION OF HERPES EYE DISEASE

疱疹性眼病复发

• 批准号: 2888527
• 负责人: DAVID J. FINK
• 金额: $ 16.78万
• 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 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2888527
• 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）从潜伏期 三叉神经节中神经元的感染是 失明 为了明确HSV的分子机制， 从潜伏期重新激活，我们建议研究的假设， HSV潜伏期的建立反映了细胞间的特异性相互作用， HSV中单个神经元和启动子元件的表型 基因组，以及病毒从潜伏期重新激活导致 神经元基因表达的可识别的改变。 概述了三个具体目标。 (1)为了定义细胞表型 含有潜伏病毒基因组的神经元， 潜伏基因组表达可检测的LAT。 私家侦探会用双- 标记免疫细胞化学、原位杂交和原位PCR技术， 神经节的连续1 μ m切片，以将细胞表征为潜伏HSV 基因组按神经递质表型，高亲和力生长的表达 因子受体和转录调控元件的存在。 (2)为了确定负责HSV再激活的机制， 在小区特定的基础上的延迟。 他将使用类似的方法后， 重新激活含有最早重新激活基因组的病毒。 (3)为了经验性地测试神经生长因子和细胞间质的作用， 早期基因c-fos和c-jun重新激活，他将构建一种病毒 用由ICP 0启动子驱动的NGF基因在细胞中早期表达NGF， 再激活级联反应，以测试NGF表达是否阻断 重新激活，并将构建缺乏环状 AMP反应元件（CRE）在ICP 0启动子中，以测试是否 细胞c-fos或c-jun表达升高激活ICP 0 导致潜伏基因组进入裂解周期的表达 活动 描述性研究将定义携带这些基因的细胞的表型。 潜伏基因组和早期发生的表型改变 重新激活级联中。 病毒构建体将根据经验测试 特定元素的潜在作用。 这些研究将 允许使用，以更好地了解细胞之间的相互作用， 病毒元件，导致潜伏基因组的重新激活， 三叉神经节;一个有重要意义的理解 防止病毒从潜伏期重新激活的策略， 失明是由这种重新激活引起的。