CELLULAR CONTROL OF CORNEAL COLLAGENASE PRODUCTION

这项建议的四个主要目标是：1)审查表格 (分泌的和/或膜结合的)其中的效应细胞(角膜 上皮细胞向靶(角膜基质)细胞提供细胞因子；2) 为了确定靶细胞上是否存在特定的受体和 细胞因子结合的特征；3)检查细胞因子结合的方式 靶细胞向其邻近底物呈递胶原酶；4) 量化和比较效应器、受体、 和胶原酶在角膜发育、生长、修复和溃疡中的作用 在效应器介导的胶原酶诱导中具有相同参数的体内 和体外抑制作用。 细胞因子的纯化将通过各种不同的 目前使用的层析和电泳法以及亲和力 结合单抗的柱层析纯化。标记细胞因子 将用于检查细胞位置和与目标细胞的结合 底物。非洲爪哇的卵母细胞系统将用来确定 以及细胞因子的细胞定位通过已经在 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.