Amelioration of recurrent herpes keratitis

描述(申请人提供)：疱疹间质角膜炎(HSK)是美国传染性失明的主要原因。视力丧失最常见的原因是复发的间质疾病，而不是原发的HSK，而且似乎是免疫介导的角膜破坏的结果。大多数实验模型都集中在原发性角膜炎上，而不是复发性角膜炎上。我们研究了近交系NIH小鼠复发HSK的模型，该模型模仿了人类复发HSK的许多临床和免疫学特征。虽然目前的资料表明原发和复发的HSK相似，但在临床病理、病毒抗原分布和角膜细胞浸润以及疫苗治疗反应方面存在显著差异，提示这两种疾病的免疫反应不同。在过去3年多的资助中，我们报告了：1)IL-1和TNFpha是复发疾病所必需的。2)复发单纯疱疹病毒性角膜炎小鼠角膜中存在Th1/Th2混合细胞群。3)复发的HSK对CD4和CD8T细胞有不同的需求，这取决于小鼠的品系。4)我们可以通过接种突变的HSV-1病毒来成功地改善疾病。基于我们的结果和从其他工作中所知道的，我们建议进一步检验复发的HSK主要是由Th1表型的CD4+T细胞介导的假设。为了验证这一假说，我们将：(1)确定IL-6和趋化因子在复发疾病中所起的作用。(2)确定需要哪些共刺激相互作用来激活和重新刺激介导复发HSK的细胞，以及针对这些相互作用的治疗干预是否将改善疾病。(3)表征在(预防性)和(治疗性)感染野生型HSV-1之前和之后用HSV-1突变株接种小鼠所产生的免疫反应，以确定这种免疫反应是否涉及Th2细胞的选择性刺激。并确定接种疫苗是否减少了潜伏感染的神经元的数量。(4)用表达OVA的HSV-1和OVA特异性TCR转基因T细胞(DO-11.10、OT-I、OT-II)建立小鼠模型，研究病毒假抗原特异性T细胞的活化、迁移、细胞因子谱和可能的耐受机制。从这些研究中获得的信息将有助于更好地理解小鼠复发的HSK的生物学机制，进而进一步了解人类疾病。此外，这些研究可能会提出更有效的免疫疗法，旨在改善人类HSK疾病。

DESCRIPTION (provided by applicant): Herpetic stromal keratitis (HSK) is a leading cause of infectious blindness in the United States. Visual loss most commonly results from recurrent stromal disease, as opposed to primary HSK and appears to be the result of immune-mediated corneal destruction. Most experimental models have focused on primary rather than recurrent keratitis. We study a model of recurrent HSK in inbred NIH mice that mimics many clinical and immunological features of recurrent HSK in humans. While current data indicates that primary and recurrent HSK are similar, there are significant differences in the clinical pathology, viral antigen distribution and cellular infiltration within the cornea, and responses to vaccine therapy suggesting that the immune responses in these two diseases is not identical. During the past 3+ years of the current grant we have reported: 1) That IL- 1 and TNFalpha are necessary for recurrent disease. 2) That there is a mixed population of Th1 and Th2 cells present in the corneas of mice with recurrent HSK. 3) That recurrent HSK has variable requirements for CD4 and CD8 T cells that depend on mouse strain. 4) That we can successfully ameliorate disease by vaccination with mutant HSV-1 viruses. Based on our results and what is known from others work we propose to further test the hypothesis that recurrent HSK is primarily mediated by CD4+ T cells of the Th1 phenotype. In order to test this hypothesis we will: (1) Determine the role that IL-6 and chemokines play in recurrent disease. (2) Determine which co stimulatory interactions are needed to activate and restimulate the cells that mediate recurrent HSK and whether therapeutic intervention targeting these interactions will ameliorate disease. (3) Characterize the immune response generated by vaccination with mutant strains of HSV-1 in mice both prior to (prophylactic) and after (therapeutic) infection with wild-type HSV-1 with an eye towards determining if that immune response involves the selective stimulation of Th2 cells. And to determine whether vaccination reduces the number of latently infected neurons. (4) Develop a mouse model using ovalbumin (OVA) expressing strains of HSV-1 and OVA-specific TCR transgenic T cells (DO-11.10, OT-I, OT-II) to characterize the activation, migration, cytokine profile, and possible tolerization protocols of viral pseudo-antigen specific T cells. The information derived from these studies will lead to a better understanding of the biology of recurrent HSK in mice and by extension human disease. Furthermore, these studies could possibly suggest more effective immunotherapies designed to ameliorate human HSK disease.