DETECTION OF HSV1 DNA IN TEARS--PROOF OF REACTIVATION

复发的HSV-1角膜疾病是一个严重的问题，可能会导致角膜疤痕形成，进而导致视力下降或失明。人类和兔子的研究表明，病毒在活动性疾病期间广泛发生，在没有角膜疾病的情况下，会发生显著的病毒脱落。聚合酶链式反应是检测单纯疱疹病毒DNA的有效方法。然而，在潜伏感染的小鼠泪液中检测HSV-1DNA的工作却很少。对小鼠的研究很重要，因为小鼠是与人类状况相关的模型，小鼠的遗传特性可以改变，以确定HSV潜伏和重新激活的基本宿主成分。这项建议的目的是1)记录在小鼠中检测到的无症状的HSV-1自发脱落和有症状的自发再激活，以及2)识别对HSV-1在小鼠的自发和诱导再激活中起重要作用的特定的小鼠(宿主)转录因子。这些目的将通过从感染病毒的野生型小鼠和特定转录因子缺失的小鼠身上获取泪液来研究。将使用聚合酶链式反应检测和定量HSV-1 DNA。荧光显微镜将用于识别感染了含有增强型绿色荧光蛋白(EGFP)基因的病毒的小鼠眼细胞和神经细胞。这些目的将有助于了解眼部HSV-1潜伏和重新激活的过程，野生型小鼠和转录基因缺陷的小鼠可能与HSV-1潜伏和重新激活有关。

Recurrent HSV-1 corneal disease is a significant problem that can lead to corneal scarring, which in turn can lead to decreased vision or blindness. Human and rabbit studies have revealed that viral shedding occurs extensively during active disease and that significant viral shedding occurs in the absence of corneal disease. The polymerase chain reaction (PCR) is a useful method for detecting HSV DNA. However, little has been done on the detection of HSV-1 DNA in the tears of latently infected mice. The study of mice is important because the mouse is a model relevant to the human condition and the genetic properties of the mouse can be varied to identify essential host components of HSV latency and reactivation. The aims of this proposal are 1) to document the detection of asymptomatic spontaneous shedding and symptomatic spontaneous reactivation of HSV-1 in mice and 2) to identify specific mouse (host) transcription factors that are important for HSV-1 spontaneous and induced reactivation in mice. These aims will be studied by obtaining tears from virus-infected wild type mice and mice with deletions in specific transcription factors. PCR will be used to detect and quantitate HSV-1 DNA. Fluorescence microscopy will be used to identify mouse ocular and nerve cells infected by a virus containing the enhanced green fluorescent protein (EGFP) gene. These aims will contribute to understanding the course of ocular HSV-1 latency and reactivation for wild type mice and mice deficient in transcription genes that are suspected to be involved in HSV-1 latency and reactivation.