Cultured human endothelial cells elaborate neutrophil chemoattractants.
Cultured human endothelial cells elaborate neutrophil chemoattractants.
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培养的人内皮细胞可产生中性粒细胞趋化剂。
DOI：
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发表时间：
1984
期刊：
影响因子：
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通讯作者：
M. Colmerauer
中科院分区：
文献类型：
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作者：
A. Mercandetti;T. Lane;M. Colmerauer
循环中性粒细胞向急性损伤肺部的募集可能受到血管内皮细胞的调节。通过博伊登室技术测定源自人脐静脉的内皮细胞汇合单层的条件培养基的中性粒细胞趋化活性。在测试的 20 个汇合单层中，14 个 (70%) 引发了趋化活性。趋化活性的产生具有时间依赖性。在四个实验中，孵育6小时时的趋化指数为3.7+/-0.4，24小时时的趋化指数为5.0+/-0.5，约48小时时的趋化指数为6.6+/-0.6。在 1:4 至 1:8 稀释度下可检测到趋化活性。为了表征条件培养基中负责增强中性粒细胞迁移的因素，来自汇合内皮细胞单层的汇集条件培养基引发最大趋化活性（汇集样品的趋化指数= 7.46）在Biogel A-0.5m柱上进行色谱分析。对应于 35,000 道尔顿和大约 1500 道尔顿分子量的两个柱级分增强了中性粒细胞迁移。从未调节的培养基中没有观察到迁移增强的柱流出物。通过棋盘分析发现来自条件培养基的两种活性柱级分均具有趋化性。 35,000 道尔顿峰的活性通过链霉蛋白酶处理或加热（56 摄氏度 X 30 分钟）消除，但未萃取到氯仿-甲醇中。相反，1500 道尔顿峰的趋化活性可抵抗热和蛋白水解，但可萃取到氯仿-甲醇中。这些数据表明，培养的人内皮细胞产生中性粒细胞趋化剂，并且产生至少两种趋化剂，即35,000道尔顿的蛋白质和1500道尔顿的脂质。我们的研究结果表明，内皮细胞有能力将中性粒细胞募集到血管表面，从而可能引发或调节炎症反应。
The recruitment of circulating neutrophils to acutely injured lungs might be regulated by vascular endothelial cells. Conditioned media from confluent monolayers of endothelial cells derived from human umbilical veins were assayed for neutrophil chemotactic activity by the Boyden chamber technique. Among 20 confluent monolayers tested, 14 (70%) elicited chemotactic activity. The generation of chemotactic activity was time dependent. In four experiments, the chemotactic index at 6 hours of incubation was 3.7 +/- 0.4, at 24 hours was 5.0 +/- 0.5, and at approximately 48 hours was 6.6 +/- 0.6. Chemotactic activity was detectable in dilutions of 1:4 to 1:8. To characterize the factors in conditioned media responsible for enhanced neutrophil migration, pooled conditioned media from confluent endothelial cell monolayers eliciting maximal chemotactic activity (chemotactic index = 7.46 on pooled sample) were chromatographed on a Biogel A-0.5m column. Two column fractions corresponding to molecular weights of 35,000 and approximately 1500 daltons enhanced neutrophil migration. No migration-enhancing column effluents were observed from unconditioned medium. Both of the active column fractions from conditioned media were found to be chemotactic by checkerboard analysis. Activity in the 35,000-dalton peak was abolished by either treatment with Pronase or heat (56 degrees C X 30 minutes) but was not extracted into chloroform-methanol. Conversely, chemotactic activity in the 1500-dalton peak resisted heat and proteolysis but was extractable into chloroform-methanol. These data indicate that cultured human endothelial cells generate neutrophil chemoattractants, and that at least two chemoattractants, a 35,000-dalton protein and a 1500-dalton lipid, are generated. Our findings suggest that endothelial cells have the capacity to recruit neutrophils to vascular surfaces and may thereby initiate or modulate the inflammatory response.