THE ROLE OF GK IN HSV-1 INDUCED CORNEAL SCARRING

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 单纯疱疹病毒(HSV)感染是美国最常见的严重病毒性眼部感染之一，也是病毒致盲的主要原因。眼部感染HSV-1病毒可导致眼部疾病，从轻微感染到由于角膜损伤而导致的视力丧失，这被称为角膜疤痕。HSV-1诱导的角膜瘢痕形成可导致失明，因此，在发达国家，HSV-1是感染性因素导致角膜失明的主要原因。据估计，美国70%-90%的成年人具有HSV-1和/或HSV-2抗体，其中约25%出现临床症状。据估计，在美国，每年有40万到50万人因眼部HSV复发而去看医生。周期性的眼部复发会导致角膜疤痕形成永久性视力障碍。尽管导致角膜瘢痕形成的特异性免疫反应仍然存在争议，但众所周知，HSV-1诱导的角膜瘢痕形成，以及随后的HSV-1诱导的角膜失明，是由于对病毒的免疫反应。这种有害免疫反应针对的单纯疱疹病毒1型蛋白(S)尚不清楚。每位患者仅限采一次血。这项研究只评估从患者身上收集的血液，而不是治疗或诊断干预。 我们的动物研究表明，在小鼠中，眼病的严重程度与一种称为糖蛋白K(GK)的HSV-1蛋白抗体的存在有很强的相关性。在一项小型先导性研究中，我们发现3例临床HSV复发者血清中有较高的抗GK抗体，而4例无HSV-1复发史的个体血清中未检测到抗GK抗体。我们想要用更多的血清证实持续复发性疾病患者的抗GK抗体效价高于无症状患者，并检验抗GK抗体与复发临床疾病相关的假说。 由于HSV-1和HSV-2都会感染眼睛，我们现在计划在HSV-2感染者的血清中证实我们的初步研究。因此，我们与华盛顿大学的安娜·沃尔德博士建立了合作关系，以检验我们的假设。安娜·沃尔德博士同意向我们发送总共50份来自有和没有HSV-2感染史的HSV-2感染者的血清。安娜·沃尔德博士将收集样本并将样本运往CSMC。这些标本将拥有只有沃尔德博士知道的识别符，我们将无法访问华盛顿大学的链接列表。 在这项研究的第二部分，我们将观察白介素2(IL-2)和HSV-1在MS和视神经炎中的相互作用，如下所述： 多发性硬化症(MS)是一种终生慢性疾病，主要诊断为年轻人。据估计，有30万至40万人患有多发性硬化症，每周约有200例新病例被诊断出来。多发性硬化症的临床症状从相对良性到破坏性不等，因为大脑和身体其他部分之间的沟通受到干扰。虽然MS的病因和发病机制尚不清楚，但工作假说是，对中枢神经系统抗原的自身免疫是由病毒破坏等环境因素触发的。在我们的初步结果中，我们发现，通过尸检组织学检查，表达小鼠白细胞介素2(IL-2)的重组单纯疱疹病毒1型(HSV-1)病毒对小鼠的眼睛攻击导致脱髓鞘。脱髓鞘与MS有惊人的相似之处，与MS一样，脱髓鞘是局灶性的，发现于中脑至后脑区域的腹侧白质束。此外，小鼠出现一过性完全后肢瘫痪(25%)或部分后肢瘫痪(75%)。而单纯野生型HSV感染、HSV-IL-4和HSV-干扰素-g病毒(与HSV-IL-2相同，但表达IL-4或干扰素-g，而不表达IL-2)均不能引起脱髓鞘。我们的初步研究还表明，IL-2单独不会导致脱髓鞘。因此，在我们的模型中，当病毒或细胞因子单独使用时，疾病不会发生；然而，当联合使用这些药物时，会导致脱髓鞘和临床异常，类似于在MS中经常观察到的那些。与这一建议相反，已开发的许多MS动物模型通常使用病毒模型或直接自身免疫模型来启动疾病。这项提案概述了一种结合了病毒和免疫成分的脱髓鞘新模型的使用。我们现在计划用外周血在多发性硬化症患者身上测试我们的动物发现 我们的总体工作假设是，CNS中的病毒感染和IL-2的存在相结合，导致CD8+T细胞和巨噬细胞重新聚集到CNS中。激活后，髓鞘特异的自身反应性CD8+T细胞直接引起脱髓鞘，巨噬细胞通过将髓鞘片段交叉呈递给CD8+T细胞来加强脱髓鞘，并将免疫反应推向TH1反应。本研究的目的是从MS患者不同复发-缓解期的外周血中分离T细胞，并验证工作假说，从而确定可能有效地靶向预防MS的发展或恶化的分子。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Herpes simplex virus (HSV) infections are among the most frequent serious viral eye infections in the U.S. and are a major cause of viral induced blindness. Ocular infection with HSV-1 can cause eye disease ranging from mild infection to loss of vision due to damage done to the cornea which is called corneal scarring. HSV-1 induced corneal scarring can lead to blindness and as such, HSV-1 is the leading cause of corneal blindness by an infectious agent in developed countries. It is estimated that 70-90% of the adult population in the U.S. have antibodies to HSV-1 and/or HSV-2, with about 25% showing clinical symptoms. It is estimated that there are 400,000 to 500,000 doctor visits annually in the United States for recurrent ocular HSV. Periodic ocular recurrences produces permanent visual disability by corneal scarring. Although the specific immune response leading to corneal scarring remains an area of controversy, it is well established that HSV-1 induced corneal scarring, and hence subsequent HSV-1 induced corneal blindness, is due to an immune response to the virus. The HSV-1 protein(s) against which this harmful immune response is directed is not known. Blood collection in limited to one blood draw per pateint. This research only evaluate blood collected from patients rather than a therapeutic or diagnostic intervention. Our animal studies suggest that in mice there is a strong correlation between severity of the eye disease and presence of antibody to one of HSV-1 protein call glycoprotein K (gK). In a small pilot study we have shown that sera from 3 humans with clinical HSV recurrences had high anti-gK antibody, while sera from 4 individula with no history of HSV-1 recurrences had no detectable anti-gK antibody. We want to confirm with a larger number of sera that individuals with ongoing recurrent disease have higher anti-gK antibody titers than asymptomatic individuals and test the hypotheses that anti-gK antibody correlates with recurrent clinical disease. Since both HSV-1 and HSV-2 infect the eye we now plan to confirm our preliminary studies in sera from HSV-2 infected individual. Therefore, we have established a collaboration with Dr. Anna Wald of University of Washington to test our hypothesis. Dr. Anna Wald had agreed to send us a total of 50 sera from HSV-2 infected individuals with and without history of HSV-2 infection. Dr. Anna Wald will collect the samples and ship the samples to CSMC. The specimens will have identifiers that only known to Dr. Wald and we will not have any access to a linking list at University of Washington. IN THE SECOND PART OF THIS STUDY WE ARE GOING TO LOOK AT THE INTERACTION OF INTERLEUKINE-2 (IL-2) AND HSV-1 in MS AND OPTIC NEURITIS AS DESCRIBED BELOW: Multiple sclerosis (MS) is a life-log chronic disease diagnosed primarily in young adults. It is estimated that 300,000 to 400,000 people have MS, with approximately 200 new cases being diagnosed weekly. Clinical symptoms of MS range from relatively benign to devastating as communication between the brain and other parts of the body is disrupted. While the cause and pathogenesis of MS is unknown, the working hypothesis is that autoimmunity to antigens of the CNS is triggered by environmental factors such as viral destruction. In our preliminary results, we have found that ocular challenge of mice with a recombinant herpes simplex virus type 1 (HSV-1) virus expressing murine interleukine-2 (IL-2) resulted in demyelination, as determined by histological examination at autopsy. The demyelination had striking similarities to that seen in MS. As with MS, the demyelination was focal and was found in ventral lateral white matter tracts in mid-brain to hind-brain areas. In addition, the mice developed transient complete (25%) or partial (75%) hind limb paralysis. In contrast, neither wild-type (wt) HSV infection alone nor HSV-IL-4, nor HSV-IFN-g virus (identical to HSV-IL-2 but expressing IL-4 or IFN-g instead of IL-2) infection caused demyelination. Our Preliminary Studies also suggest that IL-2 alone does not cause demyelination. Thus, in our model, when the virus or cytokine is administered individually disease does not occur; however, when administered together these agents cause demyelination and clinical abnormalities similar to those often observed in MS. In contrast to this proposal, the numerous animal models for MS that have been developed, generally use either the viral model or the direct autoimmune model to initiate disease. This proposal outlines the use of a novel model for demyelination that incorporates both a viral and an immune component. We now plan to test our animal findings in individuals with MS using peripheral blood Our overall working hypothesis is that, the combination of viral infection in the CNS and presence of IL-2 results in recruitment of CD8+ T cell and macrophages into the CNS. Following activation, the myelin-specific autoreactive CD8+ T cell directly cause demyelination, with the macrophages enhancing the demyelination by cross-presenting myelin fragments to CD8+ T cell as well as pushing the immune response toward a TH1 response. The goal of this study is to isolate the T cells from peripheral blood of individuals with MS during various period of relapsing- remitting and test the working hypothesis, thereby identifying molecules that may be effectively targeted for the prevention of the development or the exacerbation of MS.