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）角膜病是全球视觉发病的主要原因，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，我们证明了CD 8 + T淋巴细胞被选择性地吸引到三叉神经节（TG）的眼分支，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CD 8 + T细胞抑制HSV-1在神经元中潜伏的再激活，部分是通过产生IFN-γ。在接下来的时间里，我们建议使用实时荧光，共聚焦成像的整个TG和TG文化，转基因小鼠品系，和重组HSV-1，表达荧光蛋白从各种病毒启动子直接探索分子相互作用，保持一个池的HSV-1特异性记忆CD 8 + T细胞在潜伏感染的TG，并介导其调节病毒基因表达的神经元。在具体目标1中，我们将确定CD 8 + T细胞可以阻断HSV-1从潜伏期再活化的机制;在具体目标2中，我们将确定在阻断HSV-1从潜伏期再活化中调节CD 8 + T细胞功能程序差异表达的因子;在Specific Aim 3中，我们将探索有助于在潜伏感染的TG中维持HSV-1特异性记忆CD 8 + T细胞库的因素。

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.