L-selectin shedding does not regulate human neutrophil attachment, rolling, or transmigration across human vascular endothelium in vitro.
L-selectin shedding does not regulate human neutrophil attachment, rolling, or transmigration across human vascular endothelium in vitro.
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DOI：
10.4049/jimmunol.158.9.4365
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发表时间：
1997-05
影响因子：
4.4
通讯作者：
J. Allport;H. Ding;A. Ager;D. Steeber;T. Tedder;F. Luscinskas
中科院分区：
文献类型：
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作者：
J. Allport;H. Ding;A. Ager;D. Steeber;T. Tedder;F. Luscinskas
目前白细胞-内皮细胞相互作用的多步粘附级联模型预测在跨内皮迁移之前白细胞表面L-选择素的损失。我们已经使用体外粘附和跨内皮迁移测定和锌依赖性金属蛋白酶抑制剂Ro 31-9790（N-2-（（2s）-[（羟基氨基甲酰基）甲基）-4-甲基戊酰基]-N-1，3-二甲基-L-缬氨酰胺）来测试该假设，该抑制剂可防止趋化因子诱导的（例如，IL-8、FMLP、C5 a、血小板活化因子）来自分离的人嗜中性粒细胞的L-选择素内切蛋白水解裂解。抑制剂和溶剂处理的中性粒细胞表现出相同的行为，在两个粘附相互作用与4-和24-h TNF-α激活的HUVEC单层流动下，（包括初始附着率，滚动速度，稳定的粘附，和迁移）和静态粘附试验。用抗L-选择素的mAb对迁移的中性粒细胞进行流式细胞术分析，结果显示，载体处理的中性粒细胞具有最小的可检测的表面L-选择素，而载体处理的中性粒细胞在其表面上保留了与维持在37 ℃的中性粒细胞相当的L-选择素水平。此外，诱导快速形状变化和同型粘附（LAM 1 -116）的L-选择素单克隆抗体在流动或静态条件下并不增强中性粒细胞迁移的速率或程度。与LAM 1-116预孵育的中性粒细胞显示出与对照抗L-选择素mAb LAM 1 -101预孵育的中性粒细胞相似的行为。总之，这些结果表明，在中性粒细胞迁移穿过内皮细胞单层之前或期间，不需要L-选择素从中性粒细胞表面脱落，并且防止表面L-选择素蛋白水解裂解不会增强或抑制嗜中性粒细胞-内皮细胞粘附相互作用。
Current models of the multistep adhesion cascade for leukocyte-endothelial interactions predict loss of L-selectin from the leukocyte surface before transendothelial migration. We have tested this hypothesis using in vitro adhesion and transendothelial migration assays and a zinc-dependent metalloproteinase inhibitor, Ro 31-9790 (N-2-((2s)-[(hydroxycarbamoyl)methyl)-4-methylvaleryl]-N-1,3 -dimethyl-L-valinamide), which prevents chemoattractant-induced (e.g., IL-8, FMLP, C5a, platelet-activating factor) L-selectin endoproteolytic cleavage from isolated human neutrophils. Inhibitor and vehicle-treated neutrophils exhibited identical behavior during both adhesive interactions with 4- and 24-h TNF-alpha-activated HUVEC monolayers under flow, (including rate of initial attachment, rolling velocities, stable adhesion, and transmigration) and in static adhesion assays. Flow cytometric analysis of transmigrated neutrophils with mAb to L-selectin revealed that vehicle treated neutrophils had minimal detectable surface L-selectin, whereas inhibitor-treated neutrophils retained comparable levels of L-selectin on their surface as neutrophils maintained at 37 degrees C. In addition, mAb to L-selectin that induce rapid shape change and homotypic adhesion (LAM1-116) did not enhance the rate or extent of neutrophil transmigration under flow or static conditions. Neutrophils preincubated with LAM 1-116 displayed similar behavior to neutrophils preincubated with the control anti-L-selectin mAb, LAM1-101. In summary, these results demonstrate that there is no requirement for L-selectin to be shed from the surface of neutrophils before, or during, their migration across endothelial monolayers, and that prevention of surface L-selectin proteolytic cleavage does not enhance or inhibit neutrophil-endothelial cell adhesive interactions.