Analysis of the roles of ICAM-1 in neutrophil transmigration using a reconstituted mammalian cell expression model: Implication of ICAM-1 cytoplasmic domain and Rho-dependent signaling pathway
Analysis of the roles of ICAM-1 in neutrophil transmigration using a reconstituted mammalian cell expression model: Implication of ICAM-1 cytoplasmic domain and Rho-dependent signaling pathway
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DOI：
10.4049/jimmunol.166.1.544
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发表时间：
2001-01-01
影响因子：
4.4
通讯作者：
Duperray, A
中科院分区：
文献类型：
--
作者：
Sans, E;Delachanal, E;Duperray, A
ICAM-1 （CD54）和纤维蛋白原（fg）之间的相互作用已被证明可以增强白细胞粘附，但其在内皮细胞连接迁移过程中的具体作用尚不清楚。为了克服内皮细胞存在多种粘附受体的问题，我们设计了稳定表达ICAM-1（中国仓鼠卵巢ICAM-1）的中国仓鼠卵巢细胞系，仅在这些细胞中转染ICAM-1就足以再现中性粒细胞粘附和转运的整个过程。这种现象是由fg-ICAM-1相互作用介导的，因为fg的消耗，以及特异性抑制ICAM-1-fg相互作用的Ab的使用（2D5），完全消除了ICAM-1表达对PMN转运的影响。此外，这种icam -1介导的迁移显然取决于fg-ICAM-1在单层上的相互作用，而不是中性粒细胞，因为PMN与单抗的预孵卵是无效的。此外，当ICAM-1细胞质结构域被删除时，PMN的迁移而不是粘附被完全消除，这表明需要细胞内的信号传导来介导fg-ICAM-1对迁移的影响。使用小gtp结合蛋白Rho的特异性抑制剂，我们已经获得证据表明该信号级联参与其中。因此，我们的研究结果清楚地表明，ICAM-1在白细胞跨细胞连接的迁移中起着关键作用，并表明这种现象不是ICAM-1表达介导的粘附增强的直接结果。
Interaction between ICAM-1 (CD54) and fibrinogen (fg) has been shown to enhance Leukocyte adhesion, but its specific role in the process of migration across endothelial cell junctions remains unclear. To overcome the problem of multiple adhesion receptors found on endothelial cells, we have engineered stable Chinese hamster ovary cell lines expressing ICAM-1 (Chinese hamster ovary ICAM-1), The transfection of ICAM-1 alone in these cells is sufficient to recapitulate the entire process of neutrophil adhesion and transmigration. This phenomenon was mediated by fg-ICAM-1 interactions, as depletion of fg, as well as the use of an Ab that specifically inhibits ICAM-1-fg interaction (2D5), completely abolished the effect of ICAM-1 expression on PMN transmigration. In addition, this ICAM-1-mediated transmigration is clearly dependent on the occurrence of fg-ICAM-1 interactions on the monolayer, and not on neutrophils, as the preincubation of the PMN with the mAb was ineffective. Furthermore, PMN transmigration, but not adhesion, is totally abolished when the ICAM-1 cytoplasmic domain is deleted, indicating that signaling inside the cell is required to mediate the fg-ICAM-1 effect on transmigration. Using a specific inhibitor of the small GTP-binding protein Rho, we have obtained evidence that this signaling cascade is involved. Thus, our results clearly show that ICAM-1 plays a key role in the migration of leukocytes across cell junctions, and indicate that this phenomenon is not a direct consequence of the enhanced adhesion mediated by the expression of ICAM-1.