CHARACTERIZATION OF THE ADHESION OF THE HUMAN MONOCYTIC CELL-LINE U937 TO CULTURED ENDOTHELIAL-CELLS
CHARACTERIZATION OF THE ADHESION OF THE HUMAN MONOCYTIC CELL-LINE U937 TO CULTURED ENDOTHELIAL-CELLS
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DOI：
10.1172/jci111810
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发表时间：
1985-01-01
影响因子：
15.9
通讯作者：
DELAMOTTE, CA
中科院分区：
文献类型：
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作者：
DICORLETO, PE;DELAMOTTE, CA
血液传播的单核细胞与血管内皮的粘附是炎症期间该白细胞浸润到血管壁或间质空间中的第一步。单核细胞在创伤愈合和动脉粥样硬化形成中的重要作用现已被广泛接受。单核细胞粘附于内皮细胞的分子间相互作用尚不清楚。为了研究这种现象，开发了一种体外系统，该系统使用人单核细胞肿瘤细胞系U937作为血源性单核细胞的模型。发现51 Cr标记的U937细胞以高亲和力粘附于来自多种来源的培养的内皮细胞（EC）。观察到与来自几个物种的平滑肌细胞或成纤维细胞的结合少得多。条件培养基和共培养实验排除了靶细胞通过分泌因子影响U937细胞结合的可能性。U937细胞与猪主动脉EC的结合在37 ℃ 30分钟后达到平衡。4.0 ℃保温90 min。C在2个温度下具有相似的结合程度。U937与内皮的结合在9-12个U937/猪主动脉EC时达到饱和，半最大结合在1.5 × 10 - 6。106 U937细胞/ml。结合的细胞在37 ℃下以20小时的半衰期解离。C. U937细胞的粘附被阻断EC与正常人单核细胞，但不与血小板，淋巴细胞，或中性粒细胞预先孵育。胰蛋白酶处理或EC的去污剂增溶抑制U937细胞结合。用牛、大鼠和猪EC观察到EC密度对单核细胞粘附的显著影响。这些细胞的汇合培养物表现出可忽略不计的U937结合，但当接种稀疏时，相同的细胞是U937细胞粘附的极好靶。此外，当用棉签损伤牛主动脉EC的汇合培养物，然后使其在37 ℃下恢复24小时时，C，发现U937细胞最容易粘附到迁移到伤口中并邻近伤口的EC，但不粘附到远离伤口边缘的汇合单层中的EC。这些后者的结果可能有影响的单核细胞的局部粘附到血管壁在体内。
Adhesion of blood-borne monocytes to the vascular endothelium is the first step in the infiltration of this leukocyte into the vessel wall or the interstitial space during inflammation. A significant role for the monocyte in both wound healing and atherogenesis is now well accepted. The molecular interactions involved in monocyte attachment to the endothelium are unknown. To study this phenomenon an in vitro system was developed that uses the human monocytic tumor cell line U937 as a model for the blood-borne monocyte. 51Cr-labeled U937 cells were found to adhere with high affinity to cultured endothelial cells (EC) from several sources. Much less binding was observed to either smooth muscle cells or fibroblasts from several species. Conditioned medium and cocultivation experiments ruled out the possibility that target cells could affect U937 cell binding by secretion of factors. Binding of U937 cells to porcine aortic EC reached equilibrium after 30 min at 37.degree. C and 90 min at 4.degree. C with similar extent of binding at the 2 temperatures. Binding of U937 to the endothelium reached saturation at 9-12 U937 per porcine aortic EC (semiconfluent) with half-maximal binding at 1.5 .times. 106 U937 cells/ml. Bound cells dissociated with a half-life of 20 h at 37.degree. C. Adhesion of U937 cells was blocked by prior incubation of EC with normal human monocytes but not with platelets, lymphocytes, or neutrophils. Trypsin treatment or detergent solubilization of EC inhibited U937 cell binding. A striking effect of EC density on monocytic cell adhesion was observed with bovine, rat, and porcine EC. Confluent cultures of these cells exhibited negligible binding of U937, but when plated sparsely, the same cells were excellent targets for U937 cell adhesion. In addition, when confluent cultures of bovine aortic EC were wounded with a cotton swab and then allowed to recover for 24 h at 37.degree. C, U937 cells were found to adhere most readily to the EC migrating into the wound and neighboring the wound but not to EC in the confluent monolayer away from the wound edge. These latter results may have implications for the focal adhesion of monocytes to the vessel wall in vivo.