CYTOKINE-SPECIFIC CONTROL OF A MAST CELL PROTEASE GENE

肥大细胞蛋白酶基因的细胞因子特异性控制

• 批准号: 2224671
• 负责人: MICHAEL F GURISH
• 金额: $ 10.98万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-12-01 至 1997-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2224671
• 关键词: DNA footprinting; animal genetic material tag; cytokine; endopeptidases; fusion gene; gel mobility shift assay; gene expression; gene induction /repression; genetic regulatory element; interleukin 3; laboratory mouse; mast cell; molecular cloning; tissue /cell culture; transcription factor; transfection

Mast cells (MC) are an important component of many inflammatory responses such as those of immediate hypersensitivity. Two classes of MC are recognized in most species but he recent finding s of multiple proteases associated with the secretory ganules of MC in both moussena and man, has provide an alternate and more specific means of identifying different populations of MC. While the substrates for the various proteases have not been fully elucidated as yet, their differential expression suggest a finely coordinated system in which different proteases may invoke different responses in different tissues. Because of the greater number of invito-derived MC types developed in the mouse over the past several years, this system has proved very amenable to delineating all of the components of MC secretory granules. Currently, eight different proteases are defined and differentially expressed in various murine MC. Specific expression of murine MC proteases can be controlled by specific cytokines which demonstrates a developmental role for these molecules. Different cytokines have been implicated in the development of the different classes of MC in both mouse and man. This proposal aims to evaluate the controlling elements involved in the regulation of one of the controlling elements involved in the regulation of one of the MC proteases, mMCP-1, bu the cytokines IL-3 il-9, IL-1o and Kit Ligand. To defined the controlling elements upstream of the mCMP-1 gene, the flanking region will be ligated to a reporter gene and the construct will be transfected into a transformed rat mucosal MC line. In addition, the controlling elements will be evaluated by the techniques of DNase-I hypersensitivity and electrophoretic mobility-shift analyses. mMCP-1 has been chosen for this evaluation for several reasons. It exhibits tissue- specific expression in one mature mast cell subtype. It is the dominant protease expressed by MC in the intestinal mucosa following infection with various parasites. It is in in a transformed immature mast cell line by exposure to the appropriate cytokine which provide a in vitro system to delineate the various regulatory element. From the information provided by the proposal, it will be possible to better understand how cytokine interact with the MC in human tissue. This understanding then, will lead to better means to control MC response in various diseases such as asthma, allergy and mastocytosis.

肥大细胞 (MC) 是许多炎症反应的重要组成部分 例如那些立即过敏的人。 MC 的两类是 在大多数物种中都得到认可，但最近发现了多种蛋白酶 与穆塞纳和人类的 MC 分泌颗粒有关， 提供一种替代的、更具体的方法来识别不同的 MC 的人群。 虽然各种蛋白酶的底物 尚未完全阐明，它们的差异表达表明 一个精细协调的系统，其中不同的蛋白酶可以调用 不同的组织有不同的反应。 由于数量较多 过去几年在小鼠中开发的体外衍生的 MC 类型 多年来，事实证明这个系统非常适合描绘所有的 MC分泌颗粒的成分。 目前，八种不同 蛋白酶在不同的小鼠 MC 中被定义和差异表达。 鼠 MC 蛋白酶的特异性表达可以通过特定的 细胞因子证明了这些分子的发育作用。 不同的细胞因子与细胞的发育有关 小鼠和人的 MC 类型不同。 该提案旨在 评估参与调节之一的控制要素 参与 MC 之一调节的控制元件 蛋白酶、mMCP-1、细胞因子 IL-3、il-9、IL-1o 和 Kit Ligand。 到 定义了 mCMP-1 基因上游的控制元件， 侧翼区域将连接到报告基因，并且构建体将 转染至转化的大鼠粘膜MC系中。 此外， 控制元件将通过 DNase-I 技术进行评估 超敏反应和电泳迁移率变化分析。 mMCP-1 有 被选为此次评估有几个原因。 它展示了组织- 在一种成熟肥大细胞亚型中特异性表达。 它是占主导地位的 感染后肠粘膜中MC表达的蛋白酶 与各种寄生虫。 它位于转化的未成熟肥大细胞中 通过暴露于适当的细胞因子来提供体外 系统来界定各种监管要素。 从信息来看 通过该提案，可以更好地理解如何 细胞因子与人体组织中的 MC 相互作用。 那么这样的理解， 将带来更好的方法来控制 MC 对各种疾病的反应，例如 如哮喘、过敏和肥大细胞增多症。