ENDOTHELIAL CELL BIOLOGY IN INFLAMMATION

炎症中的内皮细胞生物学

• 批准号: 3293351
• 负责人: EUGENE C BUTCHER
• 金额: $ 18.34万
• 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 1994-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3293351
• 关键词: CD antigens; affinity chromatography; antigens; autoimmune disorder; cell adhesion molecules; cell cell interaction; cell differentiation; cellular immunity; cytokine; disease /disorder model; embryo /fetus cell /tissue; embryogenesis; human tissue; immunocytochemistry; inflammation; laboratory mouse; laboratory rabbit; laboratory rat; leukocyte activation /transformation; leukocytes; leukopoiesis; lymph nodes; monoclonal antibody; monocyte; neutrophil; tissue /cell culture; vascular endothelium

This proposal is directed at understanding the biology of endothelial cells (EC) in inflammation, and in particular the role of EC in controlling leukocyte extravasation. We have developed a panel of monoclonal antibodies (MAbs) defining segmental and pan-EC antigens; inflammation- induced antigens expressed by postcapillary venules (PCV) involved in leukocyte extravasation; and tissue-specific antigens including "vascular addressins", organ-specific EC molecules that direct the homing of lymphocytes. Our studies will focus primarily on the addressins; and on neutrophil-, monocyte-, and lymphocyte-specific adhesion mechanisms induced on EC during inflammation. The peripheral lymph node and mucosal vascular addressins (PNAd and MAd) will be characterized structurally in conventional biochemical and lectin-binding analyses; and functionally in assays of lymphocyte binding to immunoisolated PNAd. The expression of the addressins and of other EC differentiation antigens during embryogenesis, and during the development of lymphoid tissues, will be explored immunohistologically. The expression and role of the addressins in chronic inflammation will be studied in pathologic or autoimmune inflammatory models in the mouse (NOD mice, EAE, murine cerebral malaria, arthritis); and in normal and pathologically inflamed tissues in man. MAbs produced against inflamed mouse tissues (including several MAbs already identified), or against cytokine- or LPS-activated cultured EC, will be used to identify and characterize inflammation-induced antigens expressed by PCV that support leukocyte extravasation. The involvement of these antigens in leukocyte-EC interactions will be assessed in antibody inhibition studies using a) ex vivo assays of neutrophil-, monocyte-, or lymphocyte binding to postcapillary venules (PCV) in inflamed tissues; b) conventional leukocyte homing/localization studies; and c) intravital videomicroscopy of in vivo leukocyte-EC adhesion and diapedesis. The regulation of leukocyte-specific adhesion mechanisms and of the addressins in response to cytokines and/or tissue-specific factors will be studied in vitro using cultured mouse EC or human umbilical vein endothelium. Biochemical, serologic, and functional approaches will be used to characterize and determine the functional significance of two additional EC antigens, defined by Mabs MECA-32 and HECA-452, that may be involved in lymphocyte extravasation. A long-term goal will be to identify and characterize the synovial vascular addressin. The remarkable diversity and specificity of leukocyte-EC interactions renders this an exciting model for the study of basic mechanisms of cell- cell recognition and cellular positioning in vivo. Furthermore, definition of the mechanisms by which EC's control leukocyte extravasation is critical to an understanding of inflammatory and immune responses, and may permit highly selective intervention in these processes.

这项建议旨在了解内皮细胞的生物学。 (EC)在炎症中，特别是EC在控制 白细胞渗出。我们已经开发出一组单克隆性的 定义节段性抗原和泛EC抗原的抗体；炎症- 毛细血管后小静脉(PCV)表达的诱导抗原参与 白细胞外渗；和组织特异性抗原，包括“血管 Addressins“是一种器官特异性EC分子，可引导 淋巴细胞。 我们的研究将主要集中在地址蛋白；中性粒细胞-， 内皮细胞表面单核细胞和淋巴细胞特异性黏附机制的研究 发炎。外周淋巴结和粘膜血管地址 (PNAD和MAD)将在结构上以常规方式描述 生化和凝集素结合分析；以及在功能分析中 淋巴细胞与免疫分离的PNAD结合。的表达方式 Addressins和其他EC分化抗原在胚胎发育过程中， 在淋巴组织的发育过程中，将探索 免疫组织学上。地址蛋白在慢性阻塞性肺疾病中的表达及作用 炎症将在病理性或自身免疫性炎症中进行研究 小鼠模型(NOD小鼠、EAE、小鼠脑疟疾、关节炎)； 在人的正常和病理炎症组织中也是如此。生产的单抗 针对发炎的小鼠组织(包括几个已经鉴定的单抗)， 或针对细胞因子或内毒素激活的培养内皮细胞，将用于鉴定 并对PCV表达的炎症诱导抗原进行了表征 支持白细胞外渗。这些抗原在体内的作用 将在抗体抑制研究中评估白细胞与EC的相互作用 使用a)中性粒细胞、单核细胞或淋巴细胞与 炎症组织中的毛细血管后小静脉(PCV)；b)常规白细胞 归巢/定位研究；以及c)活体视频显微镜 白细胞与内皮细胞的黏附和渗出。白细胞特异性调节 细胞因子和/或寻址蛋白的黏附机制和反应 组织特异性因子将在体外利用培养的小鼠EC或 人脐静脉内皮细胞。生化、血清学和功能性 将使用各种方法来表征和确定功能 由单抗MECA-32和MECA-32定义的另外两种EC抗原的意义 HECA-452，可能与淋巴细胞外渗有关。一个长期的 目标将是鉴定和表征滑膜血管地址蛋白。 白细胞-内皮细胞相互作用的显著多样性和特异性 这为研究细胞的基本机制提供了一个令人兴奋的模型。 体内细胞识别和细胞定位。此外，定义 EC控制白细胞外渗的机制是至关重要的 对炎症和免疫反应的了解，并可能允许 对这些过程进行高度选择性的干预。