DETECTION OF HSV1 DNA IN TEARS--PROOF OF REACTIVATION

复发性HSV-1角膜疾病是一个重要的问题，可导致角膜瘢痕形成，这反过来又可导致视力下降或失明。 人类和兔子的研究表明，病毒脱落广泛发生在活动性疾病期间，并且在没有角膜疾病的情况下发生显著的病毒脱落。 聚合酶链反应（PCR）是检测HSV DNA的有效方法。 然而，很少有人做检测HSV-1 DNA在潜伏感染的小鼠的眼泪。对小鼠的研究是重要的，因为小鼠是与人类状况相关的模型，并且小鼠的遗传特性可以变化以鉴定HSV潜伏期和再活化的基本宿主组分。 本提案的目的是1）记录小鼠中HSV-1的无症状自发脱落和有症状自发再活化的检测，2）鉴定对小鼠中HSV-1自发和诱导再活化重要的特异性小鼠（宿主）转录因子。这些目标将通过从病毒感染的野生型小鼠和特定转录因子缺失的小鼠获得泪液来研究。 PCR将用于检测和定量HSV-1 DNA。荧光显微镜将用于鉴定被含有增强型绿色荧光蛋白（EGFP）基因的病毒感染的小鼠眼细胞和神经细胞。 这些目标将有助于了解野生型小鼠和转录基因缺陷小鼠的眼部HSV-1潜伏期和再激活过程，这些基因被怀疑参与HSV-1潜伏期和再激活。

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.