IMMUNOLOGICAL FUNCTIONAL MOLECULES ON HUMAN ENDOTHELIAL CELLS AND ITS CONTRIBUTION TO ANGIOGENESIS.

我们研究了EC上不同的β1整合素在修饰Matrigel结构方面的作用，Matrigel是一种由EHS肉瘤重组的凝胶。用计算机数字分析仪定量检测β1整合素亚基抗体对Matrigel诱导的毛细血管形成的抑制作用。CD_W49B、CD29抗体呈剂量依赖性抑制作用(高达70%)，而CD_W49B、CD_W49f抗体抑制作用较弱(30%)。接下来，我们观察了培养在基质中的EC的外观。EC在基质中悬浮24小时形成延长的细胞突起，每个细胞端到端连接在一起，形成毛细血管网络。此外，在基质细胞中培养48h后，部分EC(占总数的20%)显示出被EC包围的管腔中的毛细血管单位，这可能是毛细血管形成的基本单位，尽管经CD_W49B、CD29抗体处理的EC未能形成明显的突起和…更多的中空管腔。这些现象可能说明在EC分化过程中，EC与基底膜基质的相互作用在血管生成中的重要性。利用酶消化法分离纯化人皮肤肥大细胞的方法使我们能够分析这些细胞的生化和形态特征，并与其他解剖部位的肥大细胞进行更深入的比较。在本研究中，我们用免疫组织化学方法检测了分离的成人皮肤肥大细胞(SMC)是否表达表面抗原，如整合素的黏附分子、多种白细胞抗原和新发现的高亲和力IgE受体。通过对成人皮肤的酶消化和不连续的Percoll密度梯度离心法，从1克(湿重)的组织中回收大约1x10^6个肥大细胞。阿利新蓝染色显示，50%~75%的细胞为肥大细胞。用APAAP免疫组织化学染色检测细胞表面抗原的表达。与以往检测人肺或子宫肥大细胞表面标志的报道一致，SMC表达了人类白细胞抗原I类、IgE、CD29、CD43、CD45、CD49d、CD49e、CD61和CD68，而不表达CD11b、CD23和CD49b。与以往报道的其他肥大细胞不同，SMC弱表达CD11a和CD49f，但未检测到CD11c、CD18、CD54和c-kit。SMC上存在的其他表面分子包括高亲和力的IgE受体，该受体可被两种不同的单抗检测到。我们报道，SMC具有独特的表型特征，可能与皮肤的免疫反应有关。较少

We have investigated different beta1 integrins on EC for their role in modifing the structure on/in matrigel, a gel reconstituted from EHS sarcoma. The inhibition of matrigel-induced capillary formation by antibodies against subunits of beta1 integrins was examined quantitatively by using computer digital analyzer. Antibodies to CD_w49b, CD29 caused a marked inhibition (up to 70% inhibition) in a dose dependent manner, whereas antibodies to CD_w49b, CD_w49f were less inhibitory (30% inhibition). We next examined the appearances of EC cultured in the matrigel. EC suspended in matrigel for 24hr formed extended cell processes and each cell onnected together end to end resulting capillary network. Furthermore after 48hr culture in matrigel, some of EC (20% of total) showed the capillary-unit of a lumen encircled by EC which may mimic the basic putative unit in the formation of capillaries, although those pretreated with antibodies to CD_w49b, CD29 failed to form significant processes and a … More hollow lumen. These phenomenon may illustrate the importance of EC-basement membrane matrix interaction occuring during differentiation of EC in angiogenesis.Methods for the isolation and purification ofmast cells from human skin by using enzymatic digestion permitted us to analyze the biochemical and morphological characteristics of these cells and to make more intensive comparison with mast cells from other anatomical sites. In this study, we used immunohistochemical methods to determine whether isolated adult human skin mast cells (SMC) express surface antigens, such as adhesion molecules of the integrin, a variety of leukocyte antigens, and newly identified high affinity IgE receptor. By enzymatic digestion of the adult skin and centrifugation on discontinuous percoll density gradient, approximately 1x10^6 mast cells were recovered from one gram (wet weight) of the tissue. 50%-75% of the cells in cytocentrifugation speciemens were mast cells, assessed with alcian blue staining. Using APAAP immunohistochemical staining, we examined the expression of cell surface antigens. Consistent with previous reports examining surface markers on human lung or uterine mast cells, SMC were stained HLA class I,IgE,CD29, CD43, CD45, CD49d, CD49e, CD61, and CD68, however CD11b, CD23, and CD49b were not stained. In contrast to previous reports on other mast cells, SMC expressed CD11a and CD49f weakly, but CD11c, CD18, CD54, and c-kit were not detected. Additional surface molecules present on SMC included high affinity IgE receptor whichi was detected by two different monoclonal antibodies. we reported that SMC possess unique phenotypic characteristics probably responsible for the immunological reaction in skin. Less