LATENT VIRUS INFECTION AND REACTIVATION

我们已经证明，单纯疱疹病毒1型（HSV-1）胸苷激酶 (TK)表达对于HSV潜伏期是重要的。 延迟的定义是 传染性病毒分离的一般标准 潜伏感染 为了分离病毒，必须（1） 建立潜伏感染和（2）感染性病毒可以 重新激活。 在本提案中，我们计划区分 HSV TK表达对潜伏期的建立和 潜伏病毒的重新激活。 进行这次详细的调查 我们将在一个温度下进行研究， HSV-1的TK-敏感突变体和含有确定的TK-敏感突变体的HSV-1突变体。 TK基因缺失。 还将使用以下的TK突变体进行研究： HSV-2，包括缺失突变体，以确定TK的重要性 在感染发病机制中的表达。 体内互补 确定由TK-HSV建立的潜伏期， 潜伏期，并通过原位杂交检测TK-HSV， 评估。 原位杂交方法也将用于 研究是否存在HSV特异性RNA，包括TK 潜伏期的活动。 最后，抑制和慢性化HSV的方法 将利用再活化来研究再活化过程。 这些关于HSV潜伏期的建立和 重新激活将使我们深入了解这些过程的机制。

We have shown that herpes simplex virus type 1 (HSV-1) thymidine kinase (TK) expression is important for HSV latency. Latency was defined by the usual criterion of the isolation of infectious virus during the period of latent infection. For the isolation of the virus it is necessary that (1) the latent infection be established and (2) that infectious virus can be reactivated. In the present proposal we plan to differentiate between the role of HSV TK expression for the establishment of latency and for the reactivation of the latent virus. To perform this detailed investigation of the mechanisms of latency we will perform studies with a temperature sensitive TK- mutant of HSV-1 and a mutant of HSV-1 containing a defined deletion in the TK gene. Studies will also be performed with TK mutants of HSV-2, including a deletion mutant to determine the importance of TK expression in the pathogenesis of infection. In vivo complementation to determine latency established by TK- HSV, rescue of the TK- HSV during latency, and detection of TK- HSV by in situ hybridization will be evaluated. In situ hybridization methodology will also be employed to investigate the presence of HSV specified RNA, including that for TK activity during latency. Finally, methods to inhibit and to chronize HSV reactivation will be utilized to permit study of the reactivation process. These detailed studies of the establishment of HSV latency and of reactivation will provide insights into the mechanisms of these processes.