Gene Expression in HSV-1 Latency After Corneal Infection

描述(申请人提供)：单纯疱疹病毒1型(HSV-1)是导致角膜失明的主要原因。在人类中，角膜疾病是由于HSV-1在初次感染时在宿主感觉神经节建立的潜伏(静止)状态反复重新激活所致。包括我们在内的几个实验室最近的发现，挑战了一般的假设，即在HSV-1潜伏期缺乏病毒蛋白质合成会阻止对潜伏期感染神经元的免疫检测和潜伏期状态的免疫调节。相反，这些发现导致了潜伏期的动态模型，在该模型中，许多重新激活的尝试被潜伏感染神经节中的CD8+T细胞反应先发制人地中止。我们的基本假设是，潜伏期的基因表达允许神经节细胞CD8+T细胞提供一种“及时”机制，以抑制体内病毒的产生。在这项提议中，将探索在潜伏期内抗原表达和增强免疫介导的病毒潜伏期维持的概念。特殊目的1将验证这样的假设，即基因在TG中的表达发生在潜伏期并诱导重新激活，而不产生感染性病毒和真正的晚期病毒基因。我们将获得重组HSV-1，它调节来自不同候选病毒启动子的两个荧光蛋白。小鼠将通过角膜途径感染以建立潜伏期。结合共聚焦成像技术，我们将在潜伏感染和再激活诱导的小鼠的TG中鉴定HSV-1启动子的活性。我们的模型预测，在没有晚期(伽马-2)基因和传染性病毒的情况下，特定的HSV-1启动子在小鼠潜伏期内是活跃的。在特定的目标2中，我们将检验这一假设，即CD8+T细胞对神经节的免疫渗透以及它们提供的防止重新激活的保护，可以通过潜伏感染神经元中病毒表位的持续表达来增强。将开发出在潜伏期活性启动子下表达免疫优势肽多聚体的病毒。潜伏感染这种病毒的小鼠将被评估CD8+T细胞的渗透和对诱导重新激活的抵抗力。这些研究将为疫苗的设计奠定基础，以加强对潜伏期和重新激活的免疫调节。

DESCRIPTION (provided by applicant): Herpes simplex virus type 1 (HSV-1) is a major cause of corneal blindness. In humans, corneal disease results from repeated reactivation of HSV-1 from a latent (quiescent) state that was established in the host sensory ganglia at the time of primary infection. Recent findings from several laboratories, including ours, challenge the general supposition that a lack of viral protein synthesis during HSV-1 latency precludes immune detection of latently infected neurons and immune modulation of the latent state. Rather, these findings give rise to a dynamic model of latency in which many reactivation attempts are aborted by a preemptive CD8+ T cell response in the latently infected ganglia. Our underlying hypothesis is that gene expression during latency allows ganglionic CD8+T cells to provide a 'just in time' mechanism to inhibit virus production in vivo. In this proposal, the concepts of antigen expression during latency and the enhancement of the immune-mediated maintenance of viral latency will be explored. Specific Aim 1 will test the hypothesis that gene expression occurs in the TG during latency and induced reactivation without the production of infectious virus and 'true late' viral genes. We will derive recombinant HSV-1 that regulate two fluorescent proteins from different candidate viral promoters. Mice will be infected through the corneal route to establish latency. In conjunction with confocal imaging techniques, we will identify HSV-1 promoter activities in the TG of latently infected and reactivation-induced mice. Our model predicts that specific HSV-1 promoters are active during murine latency in the absence of late (gamma-2) genes and infectious virus. In Specific Aim 2, we will test the hypothesis that immune infiltration of CD8+ T cells into the ganglia, and the protection they afford from reactivation, can be enhanced by persistent expression of viral epitopes in latently infected neurons. Viruses will be developed which express multimers of an immunodominant peptide under latency active promoters. Mice latently infected with such viruses will be assessed for CD8+ T cell infiltration and resistance to induced reactivation. Such studies will establish foundations for the design of vaccines to augment immune regulation of latency and reactivation.