The heterodimer interleukin (IL)-17A/F is elevated in the lungs in chronic respiratory disease such as severe asthma, along with the pro-inflammatory cytokine tumor necrosis factor-α (TNF-α). Although IL-17A/F and TNF-α are known to functionally cooperate to exacerbate airway inflammation, proteins altered by their interaction in the lungs are not fully elucidated. We used Slow Off-rate Modified Aptamer-based proteomic array to identify proteins that are uniquely and/or synergistically enhanced by concurrent stimulation with IL-17A/F and TNF-α in human bronchial epithelial cells (HBEC). The abundance of 38 proteins was significantly enhanced by the combination of IL-17A/F and TNF-α, compared to either cytokine alone. Four out of seven proteins that were increased > 2-fold were those that promote neutrophil migration; host defence peptides (HDP; Lipocalin-2 (LCN-2) and Elafin) and chemokines (IL-8, GROα). We independently confirmed the synergistic increase of these four proteins by western blots and ELISA. We also functionally confirmed that factors secreted by HBEC stimulated with the combination of IL-17A/F and TNF-α uniquely enhances neutrophil migration. We further showed that PI3K and PKC pathways selectively control IL-17A/F + TNF-α-mediated synergistic production of HDPs LCN-2 and Elafin, but not chemokines IL-8 and GROα. Using a murine model of airway inflammation, we demonstrated enhancement of IL-17A/F, TNF-α, LCN-2 and neutrophil chemokine KC in the lungs, thus corroborating our findings in-vivo. This study identifies proteins and signaling mediated by concurrent IL-17A/F and TNF-α exposure in the lungs, relevant to respiratory diseases characterized by chronic inflammation, especially neutrophilic airway inflammation such as severe asthma.

中文翻译：

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IL-17A/F 和 TNF-α 的组合独特地改变支气管上皮细胞蛋白质组以增强增强中性粒细胞迁移的蛋白质

在严重哮喘等慢性呼吸系统疾病中，异二聚体白细胞介素 (IL)-17A/F 与促炎细胞因子肿瘤坏死因子-α (TNF-α) 一起在肺部升高。尽管已知 IL-17A/F 和 TNF-α 在功能上协同加剧气道炎症，但因它们在肺部相互作用而改变的蛋白质尚未完全阐明。我们使用基于慢解离率改良适体的蛋白质组学阵列来鉴定在人支气管上皮细胞 (HBEC) 中通过同时刺激 IL-17A/F 和 TNF-α 而独特和/或协同增强的蛋白质。与单独使用任一细胞因子相比，IL-17A/F 和 TNF-α 的组合显着增强了 38 种蛋白质的丰度。增加 > 2 倍的七种蛋白质中有四种是促进中性粒细胞迁移的蛋白质；宿主防御肽 (HDP; 脂质运载蛋白 2 (LCN-2) 和 Elafin) 和趋化因子（IL-8、GROα）。我们通过蛋白质印迹和 ELISA 独立证实了这四种蛋白质的协同增加。我们还在功能上证实，IL-17A/F 和 TNF-α 联合刺激 HBEC 分泌的因子独特地增强了中性粒细胞迁移。我们进一步表明，PI3K 和 PKC 通路选择性地控制 IL-17A/F + TNF-α 介导的 HDPs LCN-2 和 Elafin 的协同产生，但不控制趋化因子 IL-8 和 GROα。使用小鼠气道炎症模型，我们证明了肺中 IL-17A/F、TNF-α、LCN-2 和中性粒细胞趋化因子 KC 的增强，从而证实了我们在体内的发现。这项研究确定了由肺部同时暴露的 IL-17A/F 和 TNF-α 介导的蛋白质和信号，与以慢性​​炎症为特征的呼吸系统疾病相关，