Neutrophils induce sequential focal changes in endothelial adherens junction components: Role of elastase
Neutrophils induce sequential focal changes in endothelial adherens junction components: Role of elastase
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DOI：
10.1038/sj.mn.7800185
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发表时间：
2003-04-01
期刊：
影响因子：
2.4
通讯作者：
Kvietys, PR
中科院分区：
文献类型：
--
作者：
Ionescu, CV;Cepinskas, G;Kvietys, PR
目的：体外研究表明，多形核白细胞(PMN)通过细胞旁途径(即内皮细胞-细胞连接)穿过内皮细胞单层。方法：人脐静脉内皮细胞(HUVECs)在多孔膜上融合培养，IL-1β激活，甲酰-甲硫基-亮氨酰-苯丙氨酸促进PMN跨内皮细胞迁移。应用双重免疫荧光染色和激光扫描共聚焦显微镜观察PMN与内皮细胞的黏附相互作用(即黏附和跨单层迁移)对AJ组分血管内皮细胞钙粘附素、β-连环蛋白、α-连环蛋白和γ-连环蛋白的影响。结果：在贴壁PMN的AJ组分中，部分AJ组分染色缺失。远离HUVEC-PMN的AJ组分的粘附力未受影响。一种迭代方法表明，这四个组成部分依次从AJ消失。β-连环蛋白最先丢失。然后是VE-钙粘附素，α-连环蛋白，最后是伽马-连环蛋白。在没有中性粒细胞的情况下，VE-钙粘蛋白的交联，而不是血小板内皮细胞黏附分子-1或细胞间黏附分子-1的交联，增加了β-连环蛋白在细胞质中的积聚。在PMN跨内皮细胞迁移过程中，研究中的所有连接成分在单层穿透的直接部位都丢失了。同样，在从PMN单层的实际穿透部位移除的区域，AJ成分不受影响。结论：粘附性PMN可诱导AJs的局部、有序的解体，这种作用部分由PMN来源的弹性酶介导，并涉及AJs细胞内成分(即β-连环蛋白)的初始丢失。
Objective: In vitro studies have indicated that polymorphonuclear leukocytes (PMNs) traverse endothelial cell monolayers via the paracellular pathway (i.e., through endothelial cell-cell junctions. Herein, we assessed whether the adherens junctions (AJs) are disrupted during PMN transendothelial cell migration.Methods: Human umbilical vein endothelial cells (HUVECs) were grown to confluence on porous membranes and activated with interleukin-1beta, and PMN transendothelial migration was facilitated by formyl-methionyl-leucylphenylalanine. Using dual immunofluorescence staining and laser scanning confocal microscopy, we assessed the effects of PMN-endothelial cell adhesive interactions (i.e., adhesion to and emigration across monolayers) on the AJ components vascular endothelial (VE)-cadherin, beta-catenin, alpha-catenin, and gamma-catenin.Results: In the AJ immediately adjacent to the adherent PMN, there was a loss of staining for some of the AJ components. AJ components further away from HUVEC-PMN adhesive interactions were unaffected. An iterative approach indicated that the four components were sequentially lost from the AJ. beta-catenin was lost first. followed by VE-cadherin, alpha-catenin, and, finally, gamma-catenin. In the absence of PMNs, the cross-linking of VE-cadherin, but not platelet endothelial cell adhesion molecule-1 or intercellular adhesion molecule-1, increased the cytoplasmic accumulation of beta-catenin. During PMN transendothelial migration, all of the junctional components under study were lost at the immediate site of monolayer penetration. Again, at regions removed from the actual site of PMN penetration of the monolayers, the AJ components were unaffected. PMN-induced disorganization of the AJs was partially prevented by an elastase inhibitor.Conclusions: These findings suggest that adherent PMNs induce a localized, sequential disassembly of AJs, which is partially mediated by PMN-derived elastase and involves the initial loss of an intracellular component of AJs (i.e., beta-catenin).