Endogenous airway mucins carry glycans that bind Siglec-F and induce eosinophil apoptosis.
Endogenous airway mucins carry glycans that bind Siglec-F and induce eosinophil apoptosis.
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DOI：
10.1016/j.jaci.2014.10.027
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发表时间：
2015-05
期刊：
影响因子：
--
通讯作者：
Bochner BS
中科院分区：
文献类型：
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作者：
Kiwamoto T;Katoh T;Evans CM;Janssen WJ;Brummet ME;Hudson SA;Zhu Z;Tiemeyer M;Bochner BS
Siglec-F是在小鼠嗜酸性粒细胞上选择性表达的聚糖结合蛋白。它的参与诱导细胞凋亡，提示哮喘和其他嗜酸性粒细胞相关疾病中改善嗜酸性粒细胞增多的途径。Siglec-F在聚糖结合试验中识别唾液酸化、硫酸化聚糖，但内源性唾液糖苷配体及其糖蛋白载体在体内的身份尚不清楚。在体外和体内对来自正常和粘蛋白缺陷小鼠的肺以及来自小鼠的小鼠气管上皮细胞的Siglec-F配体的表达进行询问。蛋白质印迹和免疫细胞化学使用Siglec-F-Fc作为探针进行定向纯化，然后对识别的糖蛋白进行液相色谱-串联质谱分析。在小鼠嗜酸性粒细胞结合试验和基于流式细胞术的细胞死亡试验中检测纯化组分。我们检测到与Siglec-F结合的小鼠肺糖蛋白;结合是唾液酸依赖性的。Siglec-F结合材料的蛋白质组学分析鉴定了Muc 5 b和Muc 4。来自粘蛋白缺陷小鼠的肺的交叉亲和富集和组织化学分析指定并验证了Muc 5 b作为Siglec-F的一种糖蛋白配体的身份。纯化的粘蛋白制剂携带唾液酸化和硫酸化聚糖，与嗜酸性粒细胞结合，并在体外诱导其死亡。Muc 5 b条件性缺陷的小鼠表现出对IL-13的气管内安装的反应的过度嗜酸性炎症。这些数据确定了气道粘蛋白的先前未被识别的内源性抗炎性质，通过该性质，其聚糖可以通过与Siglec-F的结合来控制肺嗜酸性粒细胞增多症。
Siglec-F is a glycan binding protein selectively expressed on mouse eosinophils. Its engagement induces apoptosis, suggesting a pathway for ameliorating eosinophilia in asthma and other eosinophil-associated diseases. Siglec-F recognizes sialylated, sulfated glycans in glycan binding assays, but the identities of endogenous sialoside ligands and their glycoprotein carriers in vivo are unknown. Lungs from normal and mucin-deficient mice, as well as mouse tracheal epithelial cells from mice, were interrogated in vitro and in vivo for the expression of Siglec-F ligands. Western blotting and immunocytochemistry used Siglec-F-Fc as a probe for directed purification, followed by liquid chromatography-tandem mass spectrometric analysis of recognized glycoproteins. Purified components were tested in mouse eosinophil binding assays and flow cytometry-based cell death assays. We detected mouse lung glycoproteins that bound to Siglec-F; binding was sialic-acid dependent. Proteomic analysis of Siglec-F binding material identified Muc5b and Muc4. Cross-affinity enrichment and histochemical analysis of lungs from mucin-deficient mice assigned and validated the identity of Muc5b as one glycoprotein ligand for Siglec-F. Purified mucin preparations carried sialylated and sulfated glycans, bound to eosinophils and induced their death in vitro. Mice conditionally deficient in Muc5b displayed exaggerated eosinophilic inflammation in response to intratracheal installation of IL-13. These data identify a previously unrecognized endogenous anti-inflammatory property of airway mucins by which their glycans can control lung eosinophilia through engagement of Siglec-F.