Rhinoviral stimuli, epithelial factors and ATP signalling contribute to bronchial smooth muscle production of IL-33.
Rhinoviral stimuli, epithelial factors and ATP signalling contribute to bronchial smooth muscle production of IL-33.
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DOI：
10.1186/s12967-015-0645-3
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发表时间：
2015-08-29
影响因子：
7.4
通讯作者：
Uller L
中科院分区：
文献类型：
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作者：
Calvén J;Akbarshahi H;Menzel M;Ayata CK;Idzko M;Bjermer L;Uller L
来自严重哮喘患者的支气管平滑肌细胞 (BSMC) 已被证明过度表达 Th2 驱动和哮喘相关细胞因子 IL-33。然而，关于 BSMC 产生 IL-33 的相关因素知之甚少。鼻病毒 (RV) 感染会导致哮喘加重，表现出 Th2 型炎症的特征。在这里，我们研究了上皮源性培养基和病毒刺激对人 BSMC 中 IL-33 表达的影响。使用来自原代人支气管上皮细胞 (BEC)、双链 (ds)RNA、dsRNA/LyoVec 或感染 RV 的条件培养基刺激来自健康 (n = 3) 和哮喘 (n = 3) 受试者的原代人 BSMC。 BSMC 还用嘌呤能受体拮抗剂苏拉明进行预处理。通过 RT-qPCR 和蛋白质印迹分析 IL-33 表达，并测定细胞上清液中的 ATP 水平。 RV 感染和 dsRNA 对 TLR3 的激活增加了健康和哮喘 BSMC 中的 IL-33 mRNA 和蛋白质。这些作用被地塞米松抑制。使用 dsRNA/LyoVec 刺激 RIG-I 样受体也可增加 BSMC 的 IL-33 表达。 BEC 的条件培养基诱导 BSMC 表达 IL-33，dsRNA 进一步增强这种表达。 BEC 来源的培养基和病毒刺激的 BSMC 上清液表现出升高的 ATP 水平。用苏拉明阻断嘌呤能信号传导可抑制 dsRNA 和 BEC 衍生培养基诱导的 BSMC 表达 IL-33。 BSMC 的 RV 感染以及 TLR3 和 RIG-I 样受体的激活导致 IL-33 的表达和产生。上皮释放因子增加 BSMC 的 IL-33 表达，并与 dsRNA 表现出正相互作用。 BSMC IL-33 增加与 ATP 释放相关，并被苏拉明拮抗。我们认为，上皮衍生因子有助于基线 BSMC IL-33 的产生，而 BSMC 的 RV 感染及其病原体识别受体的刺激进一步增强了这种产生。
Bronchial smooth muscle cells (BSMCs) from severe asthmatics have been shown to overexpress the Th2-driving and asthma-associated cytokine IL-33. However, little is known regarding factors involved in BSMC production of IL-33. Rhinovirus (RV) infections cause asthma exacerbations, which exhibit features of Th2-type inflammation. Here, we investigated the effects of epithelial-derived media and viral stimuli on IL-33 expression in human BSMCs. Primary human BSMCs from healthy (n = 3) and asthmatic (n = 3) subjects were stimulated with conditioned media from primary human bronchial epithelial cells (BECs), double-stranded (ds)RNA, dsRNA/LyoVec, or infected with RV. BSMCs were also pretreated with the purinergic receptor antagonist suramin. IL-33 expression was analysed by RT-qPCR and western blot and ATP levels were determined in cell supernatants. RV infection and activation of TLR3 by dsRNA increased IL-33 mRNA and protein in healthy and asthmatic BSMCs. These effects were inhibited by dexamethasone. BSMC expression of IL-33 was also increased by stimulation of RIG-I-like receptors using dsRNA/LyoVec. Conditioned media from BECs induced BSMC expression of IL-33, which was further enhanced by dsRNA. BEC-derived medium and viral-stimulated BSMC supernatants exhibited elevated ATP levels. Blocking of purinergic signalling with suramin inhibited BSMC expression of IL-33 induced by dsRNA and BEC-derived medium. RV infection of BSMCs and activation of TLR3 and RIG-I-like receptors cause expression and production of IL-33. Epithelial-released factor(s) increase BSMC expression of IL-33 and exhibit positive interaction with dsRNA. Increased BSMC IL-33 associates with ATP release and is antagonised by suramin. We suggest that epithelial-derived factors contribute to baseline BSMC IL-33 production, which is further augmented by RV infection of BSMCs and stimulation of their pathogen-recognising receptors.