CHARACTERIZATION OF MEMBRANE SUGAR-SPECIFIC RECEPTORS IN CULTURED HIGH ENDOTHELIAL-CELLS FROM MOUSE PERIPHERAL LYMPH-NODES
CHARACTERIZATION OF MEMBRANE SUGAR-SPECIFIC RECEPTORS IN CULTURED HIGH ENDOTHELIAL-CELLS FROM MOUSE PERIPHERAL LYMPH-NODES
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DOI：
10.1016/0248-4900(93)90259-h
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发表时间：
1993-01-01
影响因子：
2.7
通讯作者：
KIEDA, C
中科院分区：
文献类型：
--
作者：
BIZOUARNE, N;MITTERRAND, M;KIEDA, C
对来自淋巴器官（外周淋巴结（PLN）和派尔氏淋巴结（PP））的专门高内皮细胞（HEC）进行培养，以研究和表征参与淋巴细胞识别并允许归巢的细胞表面分子。在分离和培养之前，通过移植物抗宿主（GVH）类型的反应在体内刺激细胞。由此产生的贴壁和生长细胞因其典型的外观和产生血管紧张素转换酶和因子 VIII 相关抗原的能力而被定性为内皮细胞。它们具有组织特异性内皮地址素。 MECA 79 抗原存在于从 PLN 分离的细胞上，而 MECA 367 抗原在来自 PP 的细胞上检测到。使用新糖蛋白和荧光显微镜对聚糖的表面受体进行细胞化学研究，并通过流式细胞术实验进行定量，结果表明糖受体的特异性取决于内皮细胞来源。事实上，α-L-岩藻糖基残基的糖受体由 PLN 的内皮细胞特异性表达。这些受体在活化的淋巴细胞条件培养基的作用下是可诱导的。外周淋巴结内皮细胞的进一步表征表明。它们确实在体外介导淋巴细胞的粘附。通过在新糖蛋白的帮助下进行的抑制实验来评估蛋白质-糖相互作用在此过程中的作用。当内皮细胞与α-L-岩藻糖基-BSA预孵育以及淋巴细胞与β-D-半乳糖基-BSA预孵育时，获得最佳抑制效果。伴随的抑制测定表明糖特异性受体（内源性凝集素）在内皮细胞和淋巴样细胞表面的参与，以实现识别和粘附。
The culture of specialized high endothelial cells (HEC) from lymphoid organs (peripheral lymph nodes (PLN) and Peyer's patches (PP)) was undertaken in order to study and characterize the cell surface molecules which are involved in lymphocyte recognition and allow homing. Cells were stimulated in vivo by a graft versus host (GVH) type of reaction before isolation and culture. The resulting adherent and growing cells were characterized as endothelial cells because of their typical aspect and their ability to produce angiotensin-converting enzyme and factor VIII-related antigen. They possess tissue-specific endothelial addressins. MECA 79 antigen is present on cells isolated from PLN while MECA 367 antigen is detected on cells from PP. Surface receptors for glycans were studied cytochemically using neoglycoproteins and fluorescence microscopy and quantified by flow cytometry experiments which showed that the specificity of sugar receptors depends upon endothelial cell origin. Indeed, sugar receptors for alpha-L-fucosyl residues were specifically expressed by endothelial cells from PLN. These receptors were inducible upon action of activated lymphocyte-conditioned medium. Further characterization of endothelial cells from peripheral lymph nodes indicates. that they indeed mediate adhesion of lymphocytes in vitro. The role of protein-sugar interactions in this process was assessed by inhibition experiments performed with the help of neoglycoproteins. Best inhibitory effects were obtained when endothelial cells had been preincubated with alpha-L-fucosyl-BSA and when lymphoid cells were preincubated with beta-D-galactosyl-BSA. Concomitant inhibition assays indicate the participation of sugar specific receptors - endogeneous lectins - on the surface of both endothelial and lymphoid cells to achieve recognition and adhesion.