Rolling on E- or P-selectin induces the extended but not high-affinity conformation of LFA-1 in neutrophils
Rolling on E- or P-selectin induces the extended but not high-affinity conformation of LFA-1 in neutrophils
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DOI：
10.1182/blood-2010-01-266122
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发表时间：
2010-07-29
期刊：
影响因子：
20.3
通讯作者：
Zarbock, Alexander
中科院分区：
文献类型：
--
作者：
Kuwano, Yoshihiro;Spelten, Oliver;Zarbock, Alexander
当细胞间粘附分子（ICAM）-1可用时，人血中性粒细胞在E-或P-选择素上滚动会降低其滚动速度。与小鼠中性粒细胞相似，这依赖于P-选择素糖蛋白配体1（PSGL 1）、α（L）β（2）整联蛋白、Src家族酪氨酸激酶FGR和脾酪氨酸激酶SYK。阻断磷脂酶C或p38 MAP激酶减弱，但没有取消速度降低。为了测试整合素活化表位的表达，我们采用了固定化报告分析，并开发了一种新的均相微流体报告抗体结合分析。在E-或P-选择素上滚动诱导延伸报告表位KIM 127和NKI-L16，但不诱导高亲和力报告表位单克隆抗体（mAb）24。这使得滚动中性粒细胞能够结合固定化的延伸报告物，但不结合活化报告物抗体，并允许在滚动期间结合可溶性KIM 127。我们的结论是，人类中性粒细胞滚动的E-或P-选择素诱导扩展的α（L）β（2）整合素的构象通过信号触发PSGL-1的参与。（血。2010; 116（4）：617-624）
Human blood neutrophils rolling on E-or P-selectin reduced their rolling velocity when intercellular adhesion molecule (ICAM)-1 was available. Similar to mouse neutrophils, this was dependent on P-selectin glycoprotein ligand 1 (PSGL1), alpha(L)beta(2) integrin, the Src family tyrosine kinase FGR and spleen tyrosine kinase SYK. Blocking phospholipase C or p38 MAP kinase attenuated, but did not abolish the velocity reduction. To test expression of integrin activation epitopes, we adapted an immobilized reporter assay and developed a new homogeneous microfluidics-based reporter antibody binding assay. Rolling on E-or P-selectin induced the extension reporter epitopes KIM127 and NKI-L16, but not the high affinity reporter epitope monoclonal antibody (mAb) 24. This enabled rolling neutrophils to bind to immobilized extension reporter, but not activation reporter antibodies and allowed binding of soluble KIM127 during rolling. We conclude that human neutrophil rolling on E-or P-selectin induces the extended alpha(L)beta(2) integrin conformation through signaling triggered by PSGL-1 engagement. (Blood. 2010; 116(4): 617-624)