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

人内皮细胞上的免疫功能分子及其对血管生成的贡献。

• 批准号: 04670636
• 负责人: KUBOTA Yasuo
• 金额: $ 1.34万
• 依托单位: ST.MARIANNA UNIVERSITY,SCHOOL OF MEDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670636/
• 关键词: ENDOTHELIAL; MAST CELLS; SURFACE ANTIGENS; ANGIOGENESIS; INTEGRINS; ADHERENCE; 基底膜; フェノタイプ; 接着

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

我们研究了EC上不同的β 1整合素在修饰基质胶（一种从EHS肉瘤重建的凝胶）上/中的结构中的作用。用计算机数字分析仪定量检测抗β 1整合素亚基抗体对基质胶诱导毛细血管形成的抑制作用。抗CD_w49b、CD 29的抗体以剂量依赖性方式引起显著抑制（高达70%抑制），而抗CD_w49b、CD_w49f的抗体抑制性较低（30%抑制）。接下来，我们检查了在基质胶中培养的EC的外观。EC在基质胶中悬浮24小时形成延伸的细胞突起，每个细胞首尾相连，形成毛细血管网络。此外，在基质胶中培养48小时后，部分EC（占总数的20%）显示出由EC包围的管腔的毛细血管单位，这可能模拟了毛细血管形成的基本假定单位，尽管用CD_w49b，CD 29抗体预处理的EC未能形成明显的突起， ...更多信息 中空腔这些现象可能说明了EC分化过程中EC-基底膜基质相互作用在血管生成中的重要性。用酶消化法从人皮肤中分离纯化肥大细胞的方法使我们能够分析这些细胞的生化和形态学特征，并与其他解剖部位的肥大细胞进行更深入的比较。在这项研究中，我们使用免疫组织化学方法来确定是否分离的成人皮肤肥大细胞（SMC）表达表面抗原，如粘附分子的整合素，各种白细胞抗原，和新发现的高亲和力IgE受体。通过酶消化成人皮肤并在不连续的琼脂糖密度梯度上离心，从每克（湿重）组织中回收约1 × 10^6个肥大细胞。细胞离心标本中50%~ 75%的细胞为肥大细胞。应用APAAP免疫组化染色检测细胞表面抗原的表达。与先前检查人肺或子宫肥大细胞表面标志物的报告一致，SMC染色为HLA I类、IgE、CD 29、CD 43、CD 45、CD 49 d、CD 49 e、CD 61和CD 68，但CD 11b、CD 23和CD 49 b未染色。与以往报道的其他肥大细胞相比，SMC表达CD 11 a和CD 49 f较弱，但未检测到CD 11 c、CD 18、CD 54和c-kit。SMC上存在的其他表面分子包括高亲和力IgE受体，其可通过两种不同的单克隆抗体检测到。我们报道了SMC具有独特的表型特征，可能是皮肤免疫反应的原因。少

项目成果

窪田 泰夫: "血管新生におけるラミニンドメイン由来ペプタイドおよびサイトカラシンBの作用について -培養ヒト血管内皮細胞を用いた検討-" 聖マリアンナ医科大学雑誌. (1993)

Yasuo Kubota：“层粘连蛋白结构域衍生肽和细胞松弛素 B 对血管生成的影响 - 使用培养的人血管内皮细胞进行的研究 -”圣玛丽安娜大学医学院杂志 (1993)。

Y.Kubota et.al: "Modulation of morphological differentiation of human endo thelial cells in culture by the synthetic peptide YIGSR and cyfuch lasin" Clinical & Experimental Darmatology. 18. 236-240 (1993)

Y.Kubota 等人：“合成肽 YIGSR 和 cyfuch lasin 对培养物中人内皮细胞形态分化的调节”临床

KUBOTA YASUO: "HUMAN SKIN MICROVASUCULAR ENDOTHELIAL CELLS -ITS MULTIPLE FUNCTIONS- (IN JAPANESE)" HIFUKANO RINSYO. 35. 1223-1228 (1993)

久保田康夫：“人类皮肤微血管内皮细胞 - 它的多种功能 - （日语）” HIFUKANO RINSYO。

窪田 泰夫: "皮膚肥満細胞" 皮膚科の臨床. 35. 1229-1236 (1993)

Yasuo Kubota：“皮肤肥大细胞”临床皮肤病学 35. 1229-1236 (1993)。

KUBOTA YASUO: "Modulation of Morphological Differentiation of Human Endothelial Cells in Culture by Synthetic peptide YIGSR and GytochabsinB." Clincal and Experimental Dermatology. (1993)

KUBOTA YASUO：“通过合成肽 YIGSR 和 GytochabsinB 调节培养中人内皮细胞的形态分化。”