ENDOTHELIAL CELL BIOLOGY IN INFLAMMATION

炎症中的内皮细胞生物学

• 批准号: 3293349
• 负责人: EUGENE C BUTCHER
• 金额: $ 15.95万
• 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/3293349
• 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.

这项建议是针对了解内皮细胞的生物学 (EC)尤其是EC在控制炎症中的作用 白细胞外渗。 我们开发了一组单克隆抗体， 定义节段性和泛EC抗原的抗体（MAb）;炎症- 毛细血管后微静脉（PCV）表达的诱导抗原参与了 白细胞外渗;和组织特异性抗原，包括“血管 地址素”是器官特异性EC分子，可指导细胞归巢 淋巴细胞 我们的研究将主要集中在地址素;和中性粒细胞， 单核细胞和淋巴细胞特异性粘附机制诱导EC期间 炎症 外周淋巴结和粘膜血管地址素 (PNAd和MAd）将在结构上以常规的 生物化学和凝集素结合分析;和功能性的测定， 淋巴细胞与免疫分离的PNAd结合。 的表达 地址素和其他EC分化抗原在胚胎发生过程中， 在淋巴组织的发展过程中， 免疫组织学。 地址素在慢性胰腺炎中的表达及作用 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、鼠脑型疟疾、关节炎）; 以及人类正常和病理性炎症组织中。 针对发炎的小鼠组织（包括已经鉴定的几种MAb）， 或针对细胞因子或LPS激活的培养EC，将用于鉴定 并表征PCV表达的炎症诱导抗原， 支持白细胞外渗。 这些抗原参与了 将在抗体抑制研究中评估白细胞-EC相互作用 使用a）离体测定中性粒细胞、单核细胞或淋巴细胞与 发炎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;和c）活体视频显微镜检查， 白细胞-EC粘附和渗出。 调节白细胞特异性 粘附机制和地址素对细胞因子的应答，和/或 组织特异性因子将在体外使用培养的小鼠EC或 人脐静脉内皮细胞 生化、血清学和功能 方法将用于表征和确定功能 两种额外的EC抗原的意义，由MECA-32单克隆抗体和 HECA-452，可能参与淋巴细胞外渗。 长期 我们的目标将是确定和表征滑膜血管地址素。 白细胞-EC相互作用的显著多样性和特异性 使其成为研究细胞基本机制的令人兴奋的模型， 细胞识别和体内细胞定位。 此外，定义 EC控制白细胞外渗的关键机制 对炎症和免疫反应的理解，并可能允许 高度选择性地干预这些过程。