ENDOTHELIAL CELL BIOLOGY IN INFLAMMATION

炎症中的内皮细胞生物学

• 批准号: 3293347
• 负责人: EUGENE C BUTCHER
• 金额: $ 12.55万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-01-01 至 1989-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3293347
• 关键词: antigens; atherosclerosis; cell biology; cell cell interaction; cytokine; human tissue; inflammation; interleukin 1; laboratory mouse; laboratory rabbit; laboratory rat; leukocyte activation /transformation; leukocytes; monoclonal antibody; monocyte; monokines; tissue /cell culture; vascular endothelium

This proposal focuses on the biology of endothelial cells (EC), with the goal of understanding their differentiation in relation to inflammatory stimuli, and their role in supporting the adhesion and extravasation of leukocytes. Since our ability to study EC differentiation is currently limited by a paucity of differentiation markers, particularly in animal models, one important aim will be to use in vivo and in vitro EC isolates to generate monoclonal antibodies (MAbs) defining several classes of endothelial differentiation antigens (inflammation-associated, organ-specific, subset-specific, etc.---identified immunohistologically), whose biochemistry, regulation and (if possible) function will be characterized subsequently in vitro and/or in vivo. We have previously shown that EC in mucosal versus non-mucosal (e.g., lymph nodes) tissues express distinct organ-specific determinants for leukocyte recognition (OSDLR) that control the adhesion and tissue-specific extravasation of circulation lymphocytes, neutrophils, and probably other leukocytes. The following tools are available to study the expression of OSDLR by endothelium: lymphoid cells bearing receptors specific for lymph node or mucosal OSD; enzymatic, polysaccharide, and mono and polyclonal antibody inhibitors of specific leukocyte receptors for OSD; and simple in vitro assays of organ-specific leukocyte-EC interactions. These tools will be applied here to dissect the nature and regulation of OSDLR. The specialized high endothelial cells (HEC) in lymphoid organs express high levels of OSDLR. An in vitro rosetting assay of lymphocyte interaction with viable HEC will be used a) to assess the active role of EC in leukocyte adhesion; and b) to explore the chemistry and regulation of OSDLR by assessing their sensitivity to various enzymatic (particularly proteolytic) and chemical treatments, and by determining whether individual EC can display both mucosal and nodal specificities simultaneously. In addition, we will ask whether the adhesion of leukocytes to cultured human EC treated with IL-1 or tumor necrosis factor (which enhance EC adhesiveness is mediated by OSDLR; and whether these or other monokines, or epithelial-derived cytokines, regulate OSDLR expression. Most importantly, we will combine serologic approaches, functional assays and biochemical isolation techniques to identify, characterize and produce MAbs against OSDLR. Finally, we will explore the specificity of OSDLR in diverse lymphoid tissues in man. These studies are important to understand the biology of endothelium and its role in normal and abnormal inflammatory and immune processes; and the pathology of endothelium in, for example, the early leukocyte-mediated events in atherosclerosis.

该提案侧重于内皮细胞（EC）的生物学， 目的是了解它们与炎性 刺激，以及它们在支持粘附和外渗中的作用， 白细胞 由于我们目前研究EC分化的能力 由于缺乏分化标志物，特别是在动物中， 模型，一个重要的目的将是使用体内和体外EC分离株 以产生定义了几种类型的单克隆抗体（MAb）， 内皮分化抗原（炎症相关， 器官特异性，亚群特异性，等等-免疫组织学鉴定）， 其生物化学、调节和（如果可能的话）功能将 随后在体外和/或体内表征。 我们先前已经表明，粘膜与非粘膜（例如，淋巴 淋巴结）组织表达不同的器官特异性白细胞决定簇 识别（OSDLR），控制粘附和组织特异性 循环淋巴细胞、中性粒细胞和可能的其他细胞外渗 白细胞 以下工具可用于研究 内皮的OSDLR：携带淋巴特异性受体的淋巴细胞 结或粘膜OSD;酶、多糖、单克隆和多克隆 OSD特异性白细胞受体的抗体抑制剂; 器官特异性白细胞-EC相互作用的体外测定。 这些工具将 在这里，我们可以应用这些方法来剖析OSDLR的性质和规则。 的 淋巴器官中的特化高内皮细胞（HEC）表达高水平的 OSDLR的水平。 淋巴细胞相互作用的体外玫瑰花结试验 a）评估EC在以下方面的积极作用： 白细胞粘附;和B）探索OSDLR的化学和调节 通过评估它们对各种酶（特别是 蛋白水解）和化学处理，并通过确定个体 EC可以同时显示粘膜和淋巴结的特异性。 在 此外，我们还将询问白细胞是否粘附到培养的人体上， 用IL-1或肿瘤坏死因子（其增强EC OSDLR介导的，以及这些或其他单核因子，或 上皮源性细胞因子，调节OSDLR表达。 最重要的是， 我们将结合联合收割机血清学方法，功能测定和生化 分离技术来鉴定、表征和产生抗 OSDLR。 最后，我们将探讨OSDLR在不同情况下的特异性。 人类的淋巴组织 这些研究对于了解内皮细胞的生物学和 它在正常和异常炎症和免疫过程中的作用; 内皮病理学，例如，早期白细胞介导的 动脉粥样硬化事件。