HUMAN AUTOIMMUNITY TO A CORNEAL STROMAL ANTIGEN

穆伦氏溃疡是一种疼痛的周围性角膜溃疡，可以 坚持不懈地向盲目进发。该病的发病机制 仍然未知，诊断是一种排除。一种自身免疫病 病因被认为与角膜特异性抗原(CO-Ag)有关 这既能激发体液免疫反应，也能激发细胞免疫反应。 了解这种疾病的自身免疫病因可能会导致 更早地认识到这种疾病，并在 制定特定的治疗方案。目标1：绘制致病基因图谱 CO-Ag蛋白中的表位。CO-Ag的氨基酸序列具有 已经确定了。与整个CO-Ag相对应的重叠多肽 蛋白质将被合成。将对每个多肽片段进行测试 对穆伦氏溃疡患者血清抗体的反应性 其他角膜周围性溃疡性疾病的患者。鉴定 疾病特异性表位的研究将导致诊断的发展 穆伦氏溃疡与其他周围性角膜的鉴别试验 疾病。目标2：检验一种假设，免疫反应是由 穆伦溃疡患者对一种感染性疾病的特定决定因素 试剂可能与疾病特异的CO-1表位序列发生交叉反应 银杏叶提取物导致角膜组织损伤和融解性疾病的机制 分子拟态。目的3：分离和鉴定一个人的c DNA 用编码序列克隆CO-Ag自身抗原。合成的 寡核苷酸探针将在已知氨基酸的基础上创建 CO-Ag的酸序列。这些探头将被用来筛查人类 表达载体构建的角膜成纤维细胞c DNA文库 UniZapXR。CO-Ag cDNA克隆将作为杂交探针用于 检测不同组织中COAG基因的表达水平 Northern印迹分析。目的4：分析细胞表型。 并用抗颗粒酶抗体检测肺炎支原体 活化细胞毒性T细胞在患者角膜组织中的表达 免疫细胞化学染色试图了解细胞毒的作用 T细胞在Mooren溃疡发病机制中的作用

Mooren's ulcer is a painful peripheral corneal ulceration that can progress relentlessly to blindness. The pathogenesis of this disease remains unknown and the diagnosis is one of exclusion. An autoimmune etiology has been suggested involving a cornea-specific antigen (CO-Ag) that incites both a humoral and cell-mediated immune response. Understanding the auto immune etiology of the disease could result in earlier recognition of the disease and be of immense value in the development of specific treatment regimens. Aim 1: To map pathogenic epitopes within the CO-Ag protein. The amino acid sequence of CO-Ag has been determined. Overlapping peptides corresponding to the entire CO-Ag protein will be synthesized. Each peptide fragment will be tested for reactivity against serum antibody from Mooren's ulcer patients and patients with other corneal peripheral ulcerative diseases. Identification of a disease-specific epitope will lead to the development of a diagnostic test to differentiate Mooren's ulcer from other peripheral corneal diseases. Aim 2: To test the hypothesis that an immune response mounted by Mooren's ulcer patient against a particular determinant of an infectious agent may cross-react with a sequence of a disease-specific epitope of CO- Ag leading to corneal tissue injury and melting disease by the mechanism of molecular mimicry. Aim 3: To isolate and characterize a human cDNA clone with coding sequences for CO-Ag autoantigen. Synthetic oligonucleotide probes will be created on the basis of the known amino acid sequence of CO-Ag. These probes will be used to screen a human corneal fibroblasts cDNA library constructed in the expression vector UniZapXR. The CO-Ag cDNA clone will be used a hybridization probe to determine the level of COAg mRNA expression in different tissues on Northern blot analysis. Aim 4: To analyze the phenotype of the infiltrating cells, and to-use anti-granzyme antibody for detection of activated cytotoxic T-cells in patient's corneal tissues by immunocytochemical staining in attempt to understand the role of cytotoxic T-cells in the pathogenesis of Mooren's ulcer.