CELLULAR CONTROL OF CORNEAL COLLAGENASE PRODUCTION

该提案的四个主要目标是： 1) 审查表格 （分泌和/或膜结合）其中效应细胞（角膜 上皮细胞）将细胞因子呈递给目标（角膜基质）细胞； 2） 确定靶细胞上是否存在特定受体以及 细胞因子结合的特征； 3) 检验方法 靶细胞将胶原酶呈递给其邻近的底物； 4) 至 定量并比较效应器、受体的水平和位置， 和胶原酶在角膜发育、生长、修复和溃疡中的作用 体内效应介导的胶原酶诱导具有相同的参数 和体外抑制作用。 细胞因子的纯化可通过多种方法完成 现在使用的色谱法和电泳法以及亲和力法 在结合的单克隆抗体的柱上纯化。 标记细胞因子 将用于检查细胞位置和与靶细胞的结合 基材。 非洲爪蟾卵母细胞系统将用于确定数量 通过已有的程序对细胞因子进行细胞定位 其他生物活性物质的文献。

The four major objectives of this proposal are: 1) To examine the form (secreted and/or membrane bound) in which effector cells (corneal epithelium) present their cytokines to target (corneal stromal) cells; 2) To determine the presence of specific receptors on the target cells and the characteristics of cytokine binding; 3) To examine the way in which the target cells present collagenase to their adjacent substrate; 4) To quantitate and compare the levels and locations of effectors, receptors, and collagenases in corneal development, growth, repair and ulceration in vivo with the same parameters in effector-mediated collagenase induction and inhibition in vitro. Purification of cytokines will be accomplished by a variety of chromatographic and electrophoretic methods now in use and by affinity purification on columns of bound monoclonal antibodies. Tagged cytokines will be used to examine cellular location and binding to target cells and substrate. The Xenopus oocyte system will be used to determine the amount and cellular location of the cytokines by procedures already in the literature for other biologically active substances.