CORNEAL INFLAMMATION AND INTERLEUKIN 10

描述(改编自申请人的摘要)：本报告的重点 项目是调查常驻角膜细胞与 免疫调节细胞因子IL-10。研究将在以下情况下进行 培养人和小鼠角膜上皮细胞和成纤维细胞。RT- 聚合酶链式反应分析将被用来准确地确定哪个常驻角膜 细胞表达IL-10受体基因。流式细胞术与免疫组织学 将用于搜索不同角膜上的蛋白质产物 单元类型。IL-10与其受体结合对角膜的影响 细胞分析将从确定是否 配体结合导致细胞因子/趋化因子mRNA和/或被抑制 蛋白质合成。我们实验室先前的RT-PCR和ELISA法研究已经 显示IL-10抑制IL-6mRNA和蛋白质的合成。这个 申请人假设IL-10抑制细胞因子的合成 基因转录水平或干扰mRNA的稳定性。 初步数据表明，人和小鼠的角膜具有 IL-10。申请者将测试常驻角膜细胞是否使 IL-10蛋白。阳性结果表明IL-10可能起作用 作为常驻角膜细胞的自我调节因子。最基本的 这些研究产生的信息可能被证明对设计新的 控制眼部炎症的方法。

DESCRIPTION (Adapted from applicant's abstract): The focus of this project is to investigate resident corneal cell interaction with the immunoregulatory cytokine IL-10. Studies will be carried out with cultured human and murine corneal epithelial cells and fibroblasts. RT- PCR analysis will be used to determine precisely which resident corneal cells express IL-10 receptor mRNA. Flow cytometry and immunohistology will be used to search for the protein product on the different corneal cell types. The consequences of IL-10 binding to its receptor on corneal cells will be analyzed from the perspective of determining whether ligand binding leads to suppressed cytokine/chemokine mRNA and/or protein synthesis. Prior RT-PCR and ELISA studies in our laboratory have shown that IL-10 suppresses IL-6 mRNA and protein synthesis. The applicant hypothesizes that IL-10 inhibits cytokine synthesis at the level of gene transcription or interferes with mRNA stability. Preliminary data indicate that human and mouse corneas possess mRNA for IL-10. The applicant will test whether resident corneal cells make IL-10 protein. Positive results would suggest that IL-10 may function as an autoregulatory factor for resident corneal cells. The basic information generated by these studies may prove useful for devising new approaches to control ocular inflammation.