ENDOTHELIAL CELL BIOLOGY IN INFLAMMATION

这项建议侧重于内皮细胞(EC)的生物学， 了解它们与炎症性疾病的区别的目标 刺激及其在支持血管黏附和渗出中的作用 白细胞。因为我们研究EC差异化的能力目前 受缺乏分化标记的限制，特别是在动物中 模型，一个重要的目标将是使用体内和体外的EC分离株 为了产生定义几种类型的单抗 内皮细胞分化抗原(炎症相关的， 器官特异性、子集特异性等-免疫组织学鉴定)， 它的生化、调节和(如果可能的话)功能将是 随后在体外和/或体内表征。 我们以前已经证明，EC在粘膜中与非粘膜中(如淋巴)中的EC 结节)组织表达不同的器官特异性白细胞决定因素 控制黏附和组织特异性的识别(OSDLR) 循环中的淋巴细胞、中性粒细胞和其他可能的细胞外渗 白细胞。以下工具可用于研究 内皮介导的OSDLR：携带淋巴特异性受体的淋巴样细胞 结节或粘膜OSD；酶、多糖、单抗和多克隆 针对OSD的特异性白细胞受体的抗体抑制剂； 器官特异性白细胞-内皮细胞相互作用的体外检测。这些工具将 在此对OSDLR的性质和规制进行了剖析。这个 淋巴器官特化高内皮细胞高表达 OSDLR的级别。淋巴细胞相互作用的体外玫瑰花环试验 具有可行性的HEC将被用来a)评估EC在 以及b)探讨OSDLR的化学和调控 通过评估它们对各种酶的敏感性(特别是 蛋白质分解)和化学处理，并通过确定个体 EC可同时显示粘膜和结节的特异性。在……里面 此外，我们还会问，白细胞与培养的人 用IL-1或肿瘤坏死因子(促进EC)治疗EC 粘附性是由OSDLR介导的；无论是这些或其他单核细胞，还是 上皮来源的细胞因子，调节OSDLR的表达。最重要的是， 我们将结合血清学方法、功能分析和生化方法 鉴定、鉴定和生产抗肿瘤单抗的分离技术 OSDLR。最后，我们将探讨OSDLR在不同领域中的特殊性 人的淋巴组织。 这些研究对于理解内皮和血管内皮细胞的生物学很重要 它在正常和异常炎症和免疫过程中的作用；以及 例如，早期白细胞介导的内皮细胞的病理学 动脉粥样硬化中的事件。

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.