Frontline Science: Pathological conditioning of human neutrophils recruited to the airway milieu in cystic fibrosis
Frontline Science: Pathological conditioning of human neutrophils recruited to the airway milieu in cystic fibrosis
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DOI：
10.1002/jlb.5hi1117-454rr
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发表时间：
2018-10-01
影响因子：
5.5
通讯作者：
Tirouvanziam, Rabindra
中科院分区：
文献类型：
--
作者：
Forrest, Osric A.;Ingersoll, Sarah A.;Tirouvanziam, Rabindra
嗜中性粒细胞向气道的募集以及它们在其中的病理调节驱动组织损伤，并且与囊性纤维化（CF）患者的肺功能丧失一致。到目前为止，这些关键过程还没有在模型中得到充分的概括，阻碍了药物的开发。在这里，我们假设，幼稚血液中性粒细胞迁移到CF气道流体在体外将诱导类似的功能适应，在体内观察到的，并提供了一个模型，以确定新的疗法。我们使用多个平台（流式细胞术、细菌杀伤和代谢测定）来表征在经上皮迁移模型中招募的血液中性粒细胞的功能特性，该模型使用来自CF受试者的气道环境作为顶端化学引诱物。与体内CF气道招募的中性粒细胞相似，体外迁移至CF气道环境的中性粒细胞显示吞噬细胞受体表达和细菌杀伤抑制，但增强颗粒释放、免疫调节功能（谷胱甘肽酶-1活化）和代谢活性，包括高Glut 1表达、糖酵解和氧化剂产生。我们还确定增强的胞饮活性作为这些细胞的一个新特征。在体外用白三烯途径抑制剂acebilustat治疗减少了中性粒细胞的数量，而代谢调节剂二甲双胍减少了代谢和氧化剂的产生，但未能恢复细菌杀伤。有趣的是，我们描述了其他炎症性气道疾病中中性粒细胞的类似病理条件。我们成功地验证了这一假设，即在体外CF患者的气道环境中招募中性粒细胞诱导了与体内观察到的病理条件相似的病理条件，为靶向治疗干预开辟了新的途径。
Recruitment of neutrophils to the airways, and their pathological conditioning therein, drive tissue damage and coincide with the loss of lung function in patients with cystic fibrosis (CF). So far, these key processes have not been adequately recapitulated in models, hampering drug development. Here, we hypothesized that the migration of naive blood neutrophils into CF airway fluid in vitro would induce similar functional adaptation to that observed in vivo, and provide a model to identify new therapies. We used multiple platforms (flow cytometry, bacteria-killing, and metabolic assays) to characterize functional properties of blood neutrophils recruited in a transepithelial migration model using airway milieu from CF subjects as an apical chemoattractant. Similarly to neutrophils recruited to CF airways in vivo, neutrophils migrated into CF airway milieu in vitro display depressed phagocytic receptor expression and bacterial killing, but enhanced granule release, immunoregulatory function (arginase-1 activation), and metabolic activities, including high Glut1 expression, glycolysis, and oxidant production. We also identify enhanced pinocytic activity as a novel feature of these cells. In vitro treatment with the leukotriene pathway inhibitor acebilustat reduces the number of transmigrating neutrophils, while the metabolic modulator metformin decreases metabolism and oxidant production, but fails to restore bacterial killing. Interestingly, we describe similar pathological conditioning of neutrophils in other inflammatory airway diseases. We successfully tested the hypothesis that recruitment of neutrophils into airway milieu from patients with CF in vitro induces similar pathological conditioning to that observed in vivo, opening new avenues for targeted therapeutic intervention.