Cytotoxic Lymphocytes and HSV-1 Corneal Lesions

描述（由申请人提供）：复发性单纯疱疹病毒1型（HSV-1）角膜疾病是世界范围内视觉疾病的主要原因，是由感觉神经元中处于潜伏（静止）状态的病毒重新激活引起的。在过去的资助期间，我们证明CD8+ T淋巴细胞被选择性地吸引到三叉神经节（TG）的眼部分支，似乎与神经元形成突触连接，并在病毒建立潜伏感染后被激活。这些发现驳斥了HSV-1在潜伏期对免疫系统不可见的观点。此外，发现HSV-1特异性CD8+ T细胞在神经元潜伏期抑制HSV-1再激活，部分是通过产生IFN-。在接下来的一段时间里，我们建议使用实时荧光和共聚焦成像全TG和TG培养物，转基因小鼠菌株和表达各种病毒启动子荧光蛋白的重组HSV-1，直接探索在潜伏感染的TG中维持HSV-1特异性记忆CD8+ T细胞池的分子相互作用，并介导它们对神经元中病毒基因表达的调节。在特异性目标1中，我们将确定CD8+ T细胞阻断HSV-1潜伏性再激活的机制；在特异性目标2中，我们将确定调节CD8+ T细胞功能程序差异表达的因子，以阻止HSV-1从潜伏期再激活；在特异性目的3中，我们将探讨在潜伏感染的TG中维持HSV-1特异性记忆CD8+ T细胞池的因素。

DESCRIPTION (provided by applicant): Recurrent herpes simplex virus type 1 (HSV-1) corneal disease, a leading cause of visual morbidity worldwide, results from reactivation of virus that is in a latent (quiescent) state in sensory neurons. During the past funding period we demonstrated that CD8+ T lymphocytes are selectively attracted to the ophthalmic branch of the trigeminal ganglion (TG), appear to form synaptic junctions with neurons, and become activated after the virus has established a latent infection. These findings refute the view that HSV-1 is invisible to the immune system during latency. Moreover, the HSV-1 specific CD8+ T cells were found to inhibit HSV-1 reactivation from latency in neurons, in part through the production of IFN-. In the next period we propose to use real-time fluorescence, and confocal imaging of whole TG and TG cultures, genetically modified mouse strains, and recombinant HSV-1 that express fluorescent proteins from a variety of viral promoters to directly explore the molecular interactions that maintain a pool of HSV-1 specific memory CD8+ T cells in the latently infected TG, and mediate their regulation of viral gene expression in neurons. In Specific Aim 1 we will identify the mechanisms by which CD8+ T cells can block HSV-1 reactivation from latency; in Specific Aim 2 we will identify factors that regulate differential expression of the functional program of CD8+ T cells in blocking HSV-1 reactivation from latency; and in Specific Aim 3 we will explore the factors that contribute to the maintenance of a pool of HSV-1 specific memory CD8+ T cells in the latently infected TG.