Sialic acid-binding immunoglobulin-like lectin 8 (Siglec-8) is an activating receptor mediating β2-integrin-dependent function in human eosinophils

JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY

卷:141

Article

Carroll, Daniela J.¹  O'Sullivan, Jeremy A.¹  Nix, David B.²  Cao, Yun¹  Tiemeyer, Michael²  Bochner, Bruce S.¹ 

[1] Northwestern Univ, Dept Med, Feinberg Sch Med, Div Allergy & Immunol, Chicago, IL 60611 USA

[2] Univ Georgia, Complex Carbohydrate Res Ctr, 220 Riverbend Rd, Athens, GA 30602 USA

关键词: Eosinophil;    Siglec-8;    beta(2)-integrin;    apoptosis;    phosphoproteomics;    reduced nicotinamide adenine dinucleotide phosphate oxidase (NADPH);    p38;    c-Jun N-terminal kinase (JNK);    Akt;   

DOI  :  10.1016/j.jaci.2017.08.013

来源: Elsevier

【 摘 要 】

Background: Siglec-8 is a CD33 subfamily cell-surface receptor selectively expressed on human eosinophils. After cytokine priming, Siglec-8 mAb or glycan ligand binding causes eosinophil apoptosis associated with reactive oxygen species (ROS) production. Most CD33-related Siglecs function as inhibitory receptors, but the ability of Siglec-8 to stimulate eosinophil ROS production and apoptosis suggests that Siglec-8 might instead function as an activating receptor. Objective: We sought to determine the role of IL-5 priming and identify the signaling molecules involved in Siglec-8 function for human eosinophils. Methods: We used an mAb and/or a multimeric synthetic sulfated sialoglycan ligand recognizing Siglec-8 in combinationwith integrin blocking antibodies, pharmacologic inhibitors, phosphoproteomics, andWestern blot analysis to define the necessity of various proteins involved in Siglec-8 function for human eosinophils. Results: Cytokine priming was required to elicit the unanticipated finding that Siglec-8 engagement promotes rapid b2-integrin-dependent eosinophil adhesion. Also novel was the finding that this adhesion was necessary for subsequent ROS production and apoptosis. Siglec-8-mediated ROS was generated through reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activation because pretreatment of eosinophils with catalase (an extracellular superoxide scavenger) or NSC 23766 (a Rac GTPase inhibitor) completely inhibited Siglec-8-mediated eosinophil apoptosis. Finally, engagement of Siglec-8 on IL-5-primed eosinophils resulted in increased phosphorylation of Akt, p38, and c-Jun N-terminal kinase 1 that was also b2-integrin dependent; pharmacologic inhibition of these kinases completely prevented Siglec-8mediated eosinophil apoptosis. Conclusions: These data demonstrate that Siglec-8 functions uniquely as an activating receptor on IL-5-primed eosinophils through a novel pathway involving regulation of b2-integrindependent adhesion, NADPH oxidase, and a subset of protein kinases.

【 授权许可】

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