Src phosphorylation of endothelial cell surface intercellular adhesion molecule-1 mediates neutrophil adhesion and contributes to the mechanism of lung inflammation.
Src phosphorylation of endothelial cell surface intercellular adhesion molecule-1 mediates neutrophil adhesion and contributes to the mechanism of lung inflammation.
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DOI：
10.1161/atvbaha.110.222208
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发表时间：
2011-06
期刊：
影响因子：
--
通讯作者：
Minshall RD
中科院分区：
文献类型：
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作者：
Liu G;Vogel SM;Gao X;Javaid K;Hu G;Danilov SM;Malik AB;Minshall RD
确定 TNFα 诱导的 Src 激活和 ICAM-1 磷酸化是否快速增加内皮细胞粘附性和 PMN 隔离，而不依赖于 ICAM-1 从头合成。小鼠肺部 TNFα 暴露 5 分钟使 125I-抗 ICAM-1 mAb 结合和 111In oxine 标记的 PMN 隔离以及 Src 激活、ICAM-1 Tyr518 磷酸化以及 pTyr518-ICAM-1 与肌动蛋白的免疫共沉淀增加 3 倍。 Nox2−/− 肺中或 Src 抑制后没有反应。在用野生型 (WT)、磷酸缺陷型 (Y518F) 或磷酸模拟型 (Y518D) 小鼠 ICAM-1 cDNA 构建体转染的 COS-7 细胞中，TNFα 增加了 YN1/1.7.4 抗 ICAM-1 mAb 与 WT-ICAM-1 结合的 Bmax，但不与 Y518F-ICAM-1 结合，表明继发于 ICAM-1 磷酸化的结合亲和力增加。 Y518D-ICAM-1 突变体的表达模拟了这种效应。 TNFα 还增加了与 F-肌动蛋白共定位的 YN1/1.7.4 mAb 标记的 WT-ICAM-1 的染色强度和细胞表面聚集，这在 Y518F-ICAM-1 中未观察到，但在 Y518D-ICAM-1 中重现。最后，与表达 Y518F ICAM-1 突变体的肺相比，小鼠肺中 ICAM-1 的过表达显着增加了 LPS 诱导的经血管白蛋白渗漏和吸入 LPS 后 2 小时和 24 小时的支气管肺泡灌洗液 PMN 计数。内皮细胞 ICAM-1 Tyr518 的 Src 依赖性磷酸化通过促进 ICAM-1 聚集诱导 PMN 粘附，我们认为 ICAM-1 聚集在炎症过程中介导 PMN 的快速相肺血管积聚。
To determine whether TNFα-induced Src activation and ICAM-1 phosphorylation rapidly increases endothelial cell adhesivity and PMN sequestration independent of de novo ICAM-1 synthesis. TNFα exposure of mouse lungs for 5 min produced a 3-fold increase in 125I-anti-ICAM-1 mAb binding and 111In oxine-labeled PMN sequestration as well as Src activation, ICAM-1 Tyr518 phosphorylation, and pTyr518-ICAM-1 co-immunoprecipitation with actin. The response was absent in Nox2−/− lungs or following Src inhibition. In COS-7 cells transfected with wild-type (WT), phospho-defective (Y518F), or phospho-mimicking (Y518D) mouse ICAM-1 cDNA constructs, TNFα increased the Bmax of YN1/1.7.4 anti-ICAM-1 mAb binding to WT-ICAM-1 but not to Y518F-ICAM-1 indicating increased binding avidity secondary to ICAM-1 phosphorylation. This effect was mimicked by expression of the Y518D-ICAM-1 mutant. TNFα also increased the staining intensity and cell surface clustering of YN1/1.7.4 mAb-labeled WT-ICAM-1 that co-localized with F-actin which was not observed with Y518F-ICAM-1 but was recapitulated with Y518D-ICAM-1. Finally, overexpression of ICAM-1 in mouse lungs significantly increased LPS-induced transvascular albumin leakage and bronchoalveolar lavage PMN counts at 2 and 24 hrs after LPS inhalation compared to lungs expressing Y518F ICAM-1 mutant. Src-dependent phosphorylation of endothelial cell ICAM-1 Tyr518 induces PMN adhesion by promoting ICAM-1 clustering which we propose mediates rapid-phase lung vascular accumulation of PMNs during inflammation.