Gene Expression in HSV-1 Latency After Corneal Infection

描述（由申请方提供）：1型单纯疱疹病毒（HSV-1）是角膜失明的主要原因。在人类中，角膜疾病是由HSV-1从原发感染时在宿主感觉神经节中建立的潜伏（静止）状态反复再活化引起的。最近的研究结果从几个实验室，包括我们的，挑战一般的假设，即缺乏病毒蛋白合成在HSV-1潜伏期排除免疫检测潜伏感染的神经元和免疫调节的潜伏状态。相反，这些发现引起了一个动态模型的潜伏期，其中许多重新激活的尝试被中止的先发制人的CD 8 + T细胞反应在潜伏感染的神经节。我们的基本假设是，潜伏期期间的基因表达允许神经节CD 8 +T细胞提供一种“及时”机制来抑制体内病毒的产生。在这个提议中，将探讨潜伏期期间抗原表达的概念和增强免疫介导的病毒潜伏期的维持。具体目标1将检验以下假设：在潜伏期和诱导再激活期间，TG中发生基因表达，而不产生感染性病毒和“真正晚期”病毒基因。我们将从不同的候选病毒启动子中获得调节两种荧光蛋白的重组HSV-1。将通过角膜途径感染小鼠以建立潜伏期。结合共聚焦成像技术，我们将确定HSV-1启动子活动的潜伏感染和再激活诱导小鼠的TG。我们的模型预测，特定的HSV-1启动子是活跃的，在小鼠潜伏期的晚期（γ-2）基因和感染性病毒的情况下。在具体目标2中，我们将测试的假设，免疫浸润的CD 8 + T细胞进入神经节，并保护他们提供的再激活，可以增强潜伏感染的神经元中的病毒表位的持续表达。将开发在潜伏活性启动子下表达免疫显性肽的多聚体的病毒。将评估潜伏感染此类病毒的小鼠的CD 8 + T细胞浸润和对诱导再活化的抗性。这些研究将为疫苗的设计奠定基础，以增强潜伏期和再激活的免疫调节。

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.