Eosinophil adhesion under flow conditions activates mechanosensitive signaling pathways in human endothelial cells
Eosinophil adhesion under flow conditions activates mechanosensitive signaling pathways in human endothelial cells
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DOI：
10.1084/jem.20041315
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发表时间：
2005-09-19
影响因子：
15.3
通讯作者：
Patel, KD
中科院分区：
文献类型：
--
作者：
Cuvelier, SL;Paul, S;Patel, KD
白细胞的迁移可以受到剪应力的影响；然而，剪应力调节白细胞迁移的机制尚不清楚。我们发现，嗜酸性粒细胞或嗜酸性细胞系与Intereukin 4刺激的内皮细胞的黏附导致内皮细胞内钙剪切依赖性增加和细胞外信号调节激酶(ERK)2的磷酸化增加，但不包括c-jun氨基末端激酶或p38丝裂原激活的蛋白激酶。涂有抗体的乳胶珠被用来表征在剪切条件下特定的内皮细胞表面分子在启动信号中的作用。我们发现，血管细胞黏附分子-1或E-选择素的结扎，而不是主要组织相容性复合体I类的结扎，都能诱导细胞因子刺激的内皮细胞中ERK2磷酸化的剪切依赖性增加。在流动条件下，肌动蛋白细胞骨架与Latrunculin A的解离阻止了ERK2的磷酸化，支持了细胞骨架在机械传感中的作用。在相同的条件下，粘着斑激酶和帕西林发生了快速的磷酸化，这表明粘着斑也参与了机械转导。最后，我们发现，在流动条件下，Rho相关蛋白激酶和钙蛋白酶在嗜酸性粒细胞随后的跨内皮细胞迁移中都起着关键作用。这些数据表明，在流动条件下，白细胞黏附分子的连接导致内皮细胞中的机械转导，这可以调节随后的白细胞运输。
Leukocyte transmigration can be affected by shear stress; however, the mechanisms by which shear stress modulates transmigration are unknown. We found that adhesion of eosinophils or an eosinophilic cell line to intereukin 4-stimulated endothelial cells led to a shear-dependent increase in endothelial cell intracellular calcium and increased phosphorylation of extracellular signal-regulated kinase (ERK) 2, but not c-Jun NH2-terminal kinase or p38 mitogen-activated protein kinase. Latex beads coated with antibodies were used to characterize the role of specific endothelial cell surface molecules in initiating signaling under shear conditions. We found that ligation of either vascular cell adhesion molecule-1 or E-selectin, but not major histocompatibility complex class I, induced a shear-dependent increase in ERK2 phosphorylation in cytokine-stimulated endothelial cells. Disassembly of the actin cytoskeleton with latrunculin A prevented ERK2 phosphorylation after adhesion under flow conditions, supporting a role for the cytoskeleton in mechanosensing. Rapid phosphorylation of focal adhesion kinase and paxillin occurred under identical conditions, suggesting that focal adhesions were also involved in mechanotransduction. Finally, we found that Rho-associated protein kinase and calpain were both critical in the subsequent transendothelial migration of eosinophils under flow conditions. These data suggest that ligation of leukocyte adhesion molecules under flow conditions leads to mechanotransduction in endothelial cells, which can regulate subsequent leukocyte trafficking.