THE ROLE OF APOPTOTIC MOLECULES IN HERPETIC STROMAL

描述(申请人提供)：我们实验室一直在研究Fas和Fas Ligand在角膜移植中的作用，并发表了几篇手稿，详细阐述了这种相互作用对同种异体角膜移植的成功和防止角膜新生血管的重要性。然而，这些凋亡分子在疱疹病毒性眼病中所起的作用仍有待充分阐明。关于这一主题的唯一报告使用了一种不会产生重大疾病的病毒-老鼠株组合。因此，凋亡分子在引起或预防HSV-1介导的眼部疾病中所起的确切作用尚不清楚。为了更清楚地了解这一情况，我们进行了初步研究，用HSV-1的Kos株和C57BL/6及其突变体与HSV-1的McKrae株感染BALB/c、BALB-LPR(Fas突变体)和BALB-GLD(Fas配体突变体)小鼠，并监测角膜疾病和眼表病毒脱落。我们的观察表明，表达Fas(CD95)基因突变的小鼠比野生型BALB/c或B6小鼠有明显更严重的HSK和眼周疾病。这些小鼠品系的GLD突变体要么与野生型(BALB/c)没有区别，要么表现出中间疾病表型。初步结果还表明，病毒在BALB-LPR小鼠周围组织中的持续时间可能略长于野生型对照组，但在BALB-GLD小鼠中也发现了类似的持续时间。提示LPR小鼠的疾病表型可能不仅仅是由于病毒的持久性，而是其他机制参与其中。为了更好地确定凋亡分子在疱疹间质角膜炎中的作用，我们建议：确定Fas和FasL在眼部疾病、神经病理保护、病毒生长和病毒潜伏期维持中的特异性贡献。其次，由于我们知道复发性眼病与原发HSK不同，我们将在HSK复发性模型中进行类似的研究，以评估Fas和FasL在这种疾病中的作用。我们还将探索用可溶性形式的Fas配体治疗小鼠的疗效，作为一种更有效地控制HSV-1引发的炎症的方法。我们相信这些研究将使我们能够更好地了解Fas-Fas配体相互作用在角膜感染性疾病中所起的作用。公共卫生相关性本申请中的研究旨在更好地了解在感染单纯疱疹病毒后免疫细胞(炎症)进入角膜的控制机制。这一点很重要，因为炎症发生得越多，对角膜的损害就越严重。此外，我们将测试一种潜在的治疗方法，旨在进一步控制并可能预防炎症。

DESCRIPTION (provided by applicant): Our laboratory has been studying the role that Fas and Fas ligand play in corneal transplantation and have published several manuscripts detailing the importance of this interaction to the success of corneal allografts and the prevention of neovascularization of the cornea. However, the role that these apoptotic molecules play in herpetic eye disease remains to be fully delineated. The only report on this subject used a virus-mouse strain combination that does not produce significant disease. Thus the precise role that apoptotic molecules play in either causing or preventing HSV-1-mediated ocular disease is not clear. In order to bring more clarity to this situation, we performed preliminary studies wherein we infected BALB/c, and BALB-lpr (Fas mutants), and BALB-gld (Fas Ligand mutants) mice with the KOS strain of HSV-1 as well as C57BL/6 and its mutants with the McKrae strain of HSV-1, and monitored both corneal disease and shedding of virus from the ocular surface. Our observations indicate that mice that express a mutation in the Fas (CD95) gene had significantly worse HSK, and periocular disease, than did either wild type BALB/c or B6 mice. Gld mutants of these mouse strains were either no different than wild-type (BALB/c) or displayed an intermediated disease phenotype. Preliminary results also suggest that virus might persist in peripheral tissues slightly longer in BALB-lpr mice than in wild type controls, but similar persistence is also noted in BALB-gld mice. Suggesting that the disease phenotype in lpr mice might is not simply due to viral persistence, but that other mechanisms are involved. In order to better define the role that apoptotic molecules have during herpetic stromal keratitis we propose: To determine the specific contributions of Fas and FasL to ocular disease, protection from neuropathology, growth of virus, and maintenance of viral latency. Secondly, since we know that recurrent eye disease is not the same as primary HSK, we will perform similar studies in a recurrent model of HSK to evaluate the role of Fas and FasL in this form of the disease. We will also explore the efficacy of treating mice with a soluble form of Fas ligand as a means of more effectively controlling HSV-1 initiated inflammation. We believe these studies will enable us to better understand the role that the Fas-Fas ligand interaction plays during infectious disease of the cornea. PUBLIC HEALTH RELEVANCE The studies in this application are designed to better understand what mechanisms are involved in controlling the entry of immune cells (inflammation) into the cornea following infection with herpes simplex virus. This is important because the more inflammation that occurs the worse will be damage to the cornea. In addition, we will test a potential therapy that is designed to further control and possibly prevent inflammation.