Virus Infections In The Eye

我们对病毒和寄生虫在眼微环境中复制时发生的各种病毒学和免疫病理过程的研究包括五个方面：(1)病毒诱导的视网膜退行性过程；(2)病毒在人类疾病中的可能作用；(3)人巨细胞病毒（CMV）感染的分子诊断与发病机制；(4)眼部疱疹病毒感染；(5)视网膜弓形虫感染。我们建立了研究视网膜退行性疾病的模型系统——实验性冠状病毒视网膜病变（ECOR）。该病毒能够在轻度视网膜血管炎症的情况下诱发急性感染。最初的视网膜损伤随后是感染性病毒的清除和进行性视网膜变性。这是第一个证明病毒诱导变性、病毒持久性、病毒诱导组织损伤的遗传易感性和病毒引发自身免疫反应的视网膜模型。我们的目标是确定病理生理机制和识别基因参与视网膜退行性疾病。在过去一年中，我们取得了以下主要成果。我们已经开始对ECOR的自身免疫成分进行表征。通过western blot分析，我们已经确定了RPE细胞和视网膜细胞表位与患有视网膜变性形式的疾病的小鼠血清反应。我们正在确定这些表位的特征。基于动物模型系统，我们也开始了评估人类视网膜退行性疾病的研究。采用免疫细胞化学染色和免疫印迹法检测不明原因视网膜病变患者的自身抗体。利用患者血清筛选大鼠视网膜cDNA表达文库，鉴定出阳性克隆。其中一个克隆对晶状体上皮衍生生长因子/转录共激活因子p52和p75/DSF70的同源性为88%。这些数据表明，在该患者中发现的针对p75/p52的抗体可能与她的视网膜病变有关。

Our studies of various virologic and immunopathologic processes that occur when viruses and parasites replicate in the ocular microenvironment comprise five areas: (1) virus induced retinal degenerative processes; (2) the possible roles of viruses in human diseases; (3) molecular diagnosis and pathogenesis of cytomegalovirus (CMV) infections in man; (4) herpesvirus infections of the eye and (5) Toxoplasma gondii infections of the retina. We have established a model system for studying retinal degenerative diseases, experimental coronavirus retinopathy (ECOR). The virus is capable of inducing an acute infection in the presence of mild retinal vascular inflammation. Initial retinal damage is followed by clearance of infectious virus and progressive retinal degeneration. This is the first retinal model to demonstrate a virus induced degeneration, viral persistence, a genetic predisposition to virus induced tissue damage and a virus triggered autoimmune response. Our goal is to determine the pathophysiological mechanisms and to identify genes involved in the retinal degenerative disease. During the past year we have made the following key findings. We have initiated a characterization of the autoimmune component in ECOR. By western blot analysis we have identified both RPE cells and retinal cell epitopes that react with sera from mice with the retinal degeneration form of the disease. We are in the process of characterizing these epitopes. Based on the animal model system, we have also initiated studies to evaluate human retinal degenerative diseases. Autoantibodies were detected in patients with retinopathy of unknown origin by immunocytochemical staining and western blot analysis. Using patient sera, a rat retina cDNA expression library was screened and positive clones were identified. One of the clones demonstrated 88% identity to lens epithelium-derived growth factor / transcription coactivators p52 and p75/DSF70. These data suggest that antibodies directed against p75/p52 identified in this patient may contribute to her retinopathy. Human CMV is a herpesvirus that is a major cause of blindness in children born with congenital infections and in immunocompromised individuals. It is difficult to study CMV latency in man. Therefore cell culture models of CMV replication and latency may provide insight into a rationale for alternative treatment modalities. Chemokines regulate leukocyte trafficking and may be important in inflammatory cell recruitment into the retina. HCMV infection in HRPE cells resulted in modulation of three important chemokines, MCP-1, MCP-3 and IL-8. The levels of MCP-1 and MCP-3 mRNA and protein in HCMV infected HRPE were reduced in comparison to uninfected HRPE. In contrast, HCMV infection enhanced IL-8 mRNA and protein levels. Because chemokines facilitate the migration and activation of leukocytes to the sites of inflammation, the modulation of chemokines production by the virus suggests a possible mechanism in immune evasion of immunopathogenesis for HCMV in retinitis and immune recovery uveitis.