Effects of human rhinovirus on epithelial barrier integrity and function in children with asthma
Effects of human rhinovirus on epithelial barrier integrity and function in children with asthma
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DOI：
10.1111/cea.13097
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发表时间：
2018-05-01
影响因子：
6.1
通讯作者：
Stick, S. M.
中科院分区：
文献类型：
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作者：
Looi, K.;Buckley, A. G.;Stick, S. M.
背景：支气管上皮紧密连接（TJ）在健康气道上皮中已被广泛评估。然而，还没有研究评估的影响，人鼻病毒（HRV）感染的表达和由此产生的屏障功能在上皮紧密连接（TJ）在儿童asthmatic.Objectives：探讨HRV感染的影响，气道上皮细胞（AECs）的气道紧密连接（TJ）的表达和屏障功能的哮喘儿童和非哮喘儿童。此外，测试的假设，屏障的完整性和功能受到损害，在更大程度上由HRV在AEC从哮喘children.Methods：初级AEC从儿童哮喘和非哮喘，分化成气液界面（ALI）文化和鼻病毒感染。通过qPCR、免疫细胞化学（ICC）、细胞内蛋白质印迹（ICW）和共聚焦显微镜评估密蛋白-1、闭合蛋白和闭合小带-1（ZO-1）的表达。屏障功能进行了评估，跨上皮电阻（TER; R-T）和通透性荧光dextrans.Results：基础钛基因表达的claudin-1和occludin显着上调哮喘儿童相比，非哮喘;然而，没有差异被视为与ZO-1。有趣的是，claudin-1，occludin和ZO-1蛋白的表达显着减少，在AEC的哮喘儿童相比，非哮喘对照组，这表明可能的转录后的固有差异。HRV感染导致非哮喘儿童的TJ和气道屏障完整性短暂分离。虽然在哮喘儿童中观察到类似的分离TJ，TJ表达的显着和持续的减少，同时在TER和增加的渗透性在哮喘children observed.Conclusion：这项研究表明，新的内在差异TJ基因和蛋白质表达的AEC的儿童与非哮喘。此外，它直接相关的关系之间的HRV感染和由此产生的分离上皮TJ，导致持续改变的屏障功能在儿童哮喘。
Background: Bronchial epithelial tight junctions (TJ) have been extensively assessed in healthy airway epithelium. However, no studies have yet assessed the effect of human rhinovirus (HRV) infection on the expression and resultant barrier function in epithelial tight junctions (TJ) in childhood asthma.Objectives: To investigate the impact of HRV infection on airway epithelial TJ expression and barrier function in airway epithelial cells (AECs) of children with and without asthma. Furthermore, to test the hypothesis that barrier integrity and function is compromised to a greater extent by HRV in AECs from asthmatic children.Methods: Primary AECs were obtained from children with and without asthma, differentiated into air-liquid interface (ALI) cultures and infected with rhinovirus. Expression of claudin-1, occludin and zonula occluden-1 (ZO-1) was assessed via qPCR, immunocytochemistry (ICC), in-cell western (ICW) and confocal microscopy. Barrier function was assessed by transepithelial electrical resistance (TER; R-T) and permeability to fluorescent dextran.Results: Basal Ti gene expression of claudin-1 and occludin was significantly upregulated in asthmatic children compared to non-asthmatics; however, no difference was seen with ZO-1. Interestingly, claudin-1, occludin and ZO-1 protein expression was significantly reduced in AEC of asthmatic children compared to non-asthmatic controls suggesting possible post-transcriptional inherent differences. HRV infection resulted in a transient dissociation of TJ and airway barrier integrity in non-asthmatic children. Although similar dissociation of TJ was observed in asthmatic children, a significant and sustained reduction in TJ expression concurrent with both a significant decrease in TER and an increase in permeability in asthmatic children was observed.Conclusion: This study demonstrates novel intrinsic differences in TJ gene and protein expression between AEC of children with and without asthma. Furthermore, it correlates directly the relationship between HRV infection and the resultant dissociation of epithelial TJ that causes a continued altered barrier function in children with asthma.