Gene Expression in HSV-1 Latency After Corneal Infection

单纯疱疹病毒1型（HSV-1）是角膜盲的主要原因。在人类中，角膜疾病 由HSV-1从宿主体内建立的潜伏（静止）状态反复再激活引起 感觉神经节在原发感染的时候。包括我们在内的几个实验室的最新发现， 挑战了一般的假设，即HSV-1潜伏期内缺乏病毒蛋白合成， 潜伏感染神经元的免疫检测和潜伏状态的免疫调节。相反，这些 研究结果产生了一个动态的延迟模型，其中许多重新激活的尝试被一个 潜伏感染神经节中的抢先CD 8 + T细胞应答。我们的基本假设是 在潜伏期期间的表达允许神经节CD 8 +T细胞提供“及时”机制， 抑制体内病毒产生。在这个提议中，抗原表达的概念在潜伏期和 将探索免疫介导的病毒潜伏期维持的增强。具体目标1将测试 假设基因表达发生在TG在潜伏期和诱导的再激活，而没有 感染性病毒和“真正晚期”病毒基因的产生。我们将获得重组HSV-1， 来自不同候选病毒启动子的两种荧光蛋白。老鼠会通过角膜被感染 路由以建立延迟。结合共聚焦成像技术，我们将识别HSV-1 潜伏感染和再激活诱导小鼠的TG中的启动子活性。我们的模型预测， 特异性HSV-1启动子在不存在晚期（3 '-2）基因的鼠潜伏期期间是有活性的， 病毒在特异性目标2中，我们将检验CD 8 + T细胞免疫浸润到 神经节，以及它们提供的防止再激活的保护，可以通过持续表达病毒 潜伏感染的神经元中的表位。将开发表达多聚体的病毒， 在潜伏活性启动子下的免疫显性肽。潜伏感染这种病毒的小鼠将 评估CD 8 + T细胞浸润和对诱导再活化的抗性。这些研究将建立 为设计疫苗以增强潜伏期和再激活的免疫调节奠定了基础。

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 (3'-2) genes and infectious virus. In Specific Aim 2, we will test the hypothesis that immune infiltration ofCD8+ 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.