Herbert Cristan, Do Kimchi, Chiu Vincent, Garthwaite Linda, Chen Yifan, Young Paul M, Traini Daniela, Kumar Rakesh K
Department of Pathology, School of Medical Sciences, UNSW Sydney, NSW 2052, Australia.
Respiratory Cellular and Molecular Biology, Woolcock Institute of Medical Research, University of Sydney, Sydney, NSW 2037, Australia.
Clin Sci (Lond). 2017 Mar 1;131(6):499-509. doi: 10.1042/CS20160939. Epub 2017 Jan 23.
Airway epithelial cells (AEC) exhibit a pro-inflammatory phenotype in patients with allergic asthma. We examined the effect of an allergic cytokine environment on the response of AEC to rhinovirus (RV), the most common trigger of acute exacerbations of asthma. Calu-3 cells, a well-differentiated human AEC line, were cultured with or without the T-helper type 2 cytokines interleukin (IL)-4 and IL-13, then stimulated with a toll-like receptor (TLR) 3 agonist (poly I:C, dsRNA) or a TLR7 agonist (imiquimod), or infected with RV 16. Expression of pro-inflammatory and antiviral mediators, and of viral pattern-recognition molecules, was assessed using nCounter assays, quantitative real-time PCR (qRT-PCR) and protein immunoassays. Both dsRNA and imiquimod stimulated expression of mRNA for whereas expression of several chemokines and antiviral response genes was induced only by dsRNA. Conversely, expression of other cytokines and growth factors was induced only by imiquimod. RV infection not only stimulated expression of the inflammation-related genes induced by dsRNA, but also of complement factor B and the novel pro-inflammatory cytokine IL-32. In the T helper type 2 (Th2) cytokine environment, several mediators exhibited significantly enhanced expression, whereas expression of interferons was either unchanged or enhanced. The allergic environment also increased expression of pattern-recognition receptors and of intercellular adhesion molecule 1, the cell surface receptor for RV. We conclude that Th2 cytokines promote increased production of pro-inflammatory mediators by AEC following infection with RV. Increased viral entry or enhanced signalling via pattern-recognition receptors could also contribute to the exaggerated inflammatory response to RV observed in allergic asthmatics.
在过敏性哮喘患者中，气道上皮细胞（AEC）呈现促炎表型。我们研究了过敏性细胞因子环境对AEC对鼻病毒（RV）反应的影响，RV是哮喘急性加重最常见的诱因。将分化良好的人AEC系Calu-3细胞在有或无2型辅助性T细胞细胞因子白细胞介素（IL）-4和IL-13的情况下进行培养，然后用Toll样受体（TLR）3激动剂（聚肌胞苷酸，双链RNA）或TLR7激动剂（咪喹莫特）刺激，或用RV 16感染。使用nCounter分析、定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫分析评估促炎和抗病毒介质以及病毒模式识别分子的表达。双链RNA和咪喹莫特均刺激了[具体分子]mRNA的表达，而几种趋化因子和抗病毒反应基因的表达仅由双链RNA诱导。相反，其他细胞因子和生长因子的表达仅由咪喹莫特诱导。RV感染不仅刺激了双链RNA诱导的炎症相关基因的表达，还刺激了补体因子B和新型促炎细胞因子IL-32的表达。在2型辅助性T细胞（Th2）细胞因子环境中，几种介质的表达显著增强，而干扰素的表达要么未改变，要么增强。过敏环境还增加了模式识别受体和细胞间黏附分子1（RV的细胞表面受体）的表达。我们得出结论，Th2细胞因子促进RV感染后AEC促炎介质产生增加。病毒进入增加或通过模式识别受体的信号增强也可能导致过敏性哮喘患者对RV的炎症反应过度。
