Cell Movement Through a TH2-Conditioned Extracellular Matrix

我们的长期目标是了解嗜酸性粒细胞（EOS）如何交通和相互作用，并有助于 哮喘的发展。aM32整联蛋白（CD 11b/CD 18）在EOS上高度活化， 节段性抗原激发后支气管肺泡灌洗，表明aM（32）功能变得 重要的是EOS渗出并在肺中迁移。然而，关于aMp 2和 相关配体参与气道EOS的粘附、迁移和活性。本报告的总体目标 本发明的目的是确定aMp 2在调节气道中EOS行为中的作用。基于我们 哮喘中EOS生物学和细胞外基质（ECM）的初步数据和当前认识， 目前的假设是骨膜蛋白，一种由辅助性T细胞2型驱动的炎症特征性ECM蛋白， 在哮喘患者中发现的EOS α Mp2是EOS α Mp2整联蛋白的主要粘附配体， aMp 2-骨膜蛋白相互作用是EOS功能的重要决定因素。目的1是定义aMp 2的作用， 骨膜蛋白和TGF-β诱导蛋白（TGFBI），一种也在肺中发现的骨膜蛋白同系物，在粘附，迁移， sun/ival和由IL-3、IL-5、GM-CSF或其它活化剂活化的EOS的其它功能。目标二是 确定了aMP 2识别骨膜蛋白的结构-功能关系，绘制了aMP 2与骨膜蛋白的结合图谱， 位点，并定义最小构建体，当固定化时支持EOS，当可溶性阻断EOS时 粘附和迁移。维生素K依赖的γ-羧基化和选择性剪接如何影响 还将测定骨膜蛋白的生物活性。目的3是了解机制， 在哮喘气道中发现骨膜蛋白显著增加的意义。骨膜蛋白分泌 成纤维细胞和上皮细胞受TGF-β或其他因子刺激，以及其沉积到ECM中， 将对营业额进行分析。将开发针对各种形式的骨膜蛋白和TGFBI的抗体， 局部抗原攻击后支气管活检中的定位。实现本提案的目标将 提供新的知识和更好地了解EOS贩运和功能及其相互作用 在Th 2驱动的炎症中，EOS和ECM之间的相互作用，并将产生允许 这种相互作用有待研究和调整。 RELE^万斯（参见说明）： 拟议的研究将确定结缔组织蛋白骨膜蛋白，这是强烈上升- 在哮喘中调节，与其细胞表面受体蛋白aMP 2整合素（CD 11b/CD 18）相互作用， 支持EOS的附着和迁移。该项目将提供新的见解 哮喘气道EOS的运动和病变肺结缔组织的生物学。

Our long-term goal is to understand how eosinophils (EOS) traffic to and interact in the ainway and contribute to the progression of asthma. aM32 integrin (CD11b/CD18) is highly activated on EOS obtained by bronchoalveolar lavage after segmental antigen challenge, suggesting that aM(32 functions becomes important as EOS extravasate to and migrate in the lung. However, little is known about roles of aMp2 and relevant ligand(s) in adhesion, migration, and activities of EOS in the airway. The overall objective of this proposal is to determine the role of aMp2 in modulating behavior of EOS in the ainway. Based on our preliminary data and current understanding of EOS biology and the extracellular matrix (ECM) in asthma, the current hypothesis is that periostin, an ECM protein characteristic of inflammation driven by T helper type 2 cells and found in the asthmatic ain/vay, is a dominant adhesive ligand for EOS aMp2 integrin and that the aMp2-periostin interaction is an important determinant of EOS function. Aim 1 is to define roles of aMp2, periostin and TGF-P-induced protein (TGFBI), a periostin homolog also found in lung, in adhesion, migration, sun/ival, and other functions of EOS activated by IL-3, IL-5, GM-CSF, or other activators. Aim 2 is to determine the structure-function relationship ofthe recognition of periostin by aMP2, map the aMp2-binding site(s), and define minimal constructs that when immobilized support, and when soluble block, EOS adhesion and migration. How the vitamin K-dependent y-carboxylation and alternative splicing affect periostin's biological activities will also be determined. Aim 3 is to understand the mechanism and significance ofthe striking increase of periostin that is found in the asthmatic airway. Periostin secretion from fibroblasts and epithelial cells stimulated by TGF-p or other factors, as well as its deposition into ECM and turnover will be analyzed. Antibodies to the various forms of periostin and TGFBI will be developed for localization in bronchial biopsies after segmental antigen challenge. Achieving the goals of this proposal will provide novel knowledge and a better understanding of EOS trafficking and functions and the interplay between EOS and the ECM in Th2-driven inflammation, and will generate agents and reagents that will allow this interplay to be studied and modulated. RELE^VANCE (See instructions): The proposed research will determine how the connective tissue protein periostin, which is strongly up- regulated in asthma, interacts with its cell-surface receptor protein aMP2 integrin (CD11b/CD18) on activated EOS, and supports attachment and migration of EOS. The project will provide new insights into the movement of EOS in the asthmatic ainway and the biology of connective tissue in the diseased lung.