CELLULAR CONTROL OF CORNEAL COLLAGENASE PRODUCTION

这项建议的四个主要目标是：1）审查形式 （分泌的和/或膜结合的），其中效应细胞（角膜上皮细胞） 上皮）将其细胞因子呈递给靶（角膜基质）细胞; 2） 为了确定靶细胞上特异性受体的存在， 细胞因子结合的特征; 3）检查细胞因子结合的方式， 靶细胞将胶原酶呈递至其相邻底物; 4） 定量并比较效应物，受体， 和胶原酶在角膜发育、生长、修复和溃疡形成中的作用。 在效应介导的胶原酶诱导中具有相同参数的体内 和体外抑制。 细胞因子的纯化将通过多种方法来完成。 目前使用的色谱和电泳方法， 在结合的单克隆抗体的柱上纯化。 Tagged细胞因子 将用于检查细胞定位和与靶细胞的结合， 衬底 非洲爪蟾卵母细胞系统将用于确定 和细胞定位的细胞因子的程序已经在 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.