Staphylococcus aureus impairs the airway epithelial barrier in vitro
Staphylococcus aureus impairs the airway epithelial barrier in vitro
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DOI：
10.1002/alr.21517
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发表时间：
2015-06-01
影响因子：
6.4
通讯作者：
Vreugde, Sarah
中科院分区：
文献类型：
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作者：
Malik, Zacki;Roscioli, Eugene;Vreugde, Sarah
背景慢性鼻窦炎（CRS）是一组导致鼻窦粘膜炎症的疾病。金黄色葡萄球菌(S.金黄色葡萄球菌）与严重和顽固性CRS相关。本研究旨在探讨S.金黄色葡萄球菌条件培养液对呼吸道上皮屏障结构和功能的影响。将金黄色葡萄球菌参考菌株（美国典型培养物保藏中心[ATCC] 13565、14458和25923）应用于原代人鼻上皮细胞（HNEC）的气-液界面（ALI）培养物，并测量跨上皮电阻（TEER）以评估细胞-细胞完整性。用电子显微镜测量纤毛区和紧密连接（TJ）。此外，通过免疫荧光检测TJ蛋白封闭带-1（ZO-1）的表达。统计学分析采用方差分析（ANOVA）和两两Bonferroni校正的t检验。与对照和参考菌株14458和25923相比，金黄色葡萄球菌菌株13565引起电阻抗的浓度依赖性下降（p < 0.001）。电子显微镜显示，在TJ区域，相邻细胞之间在顶部有明显的分离。结论S13565条件培养液对HNECs的纤毛区无明显影响，但ZO-1表达不连续。金黄色葡萄球菌菌株13565通过破坏ALI时生长的原代HNEC之间的TJ而损伤气道上皮。这些结果表明，菌株特异性S.金黄色葡萄球菌分泌的产物损害了上皮屏障功能，这可能是S.金黄色葡萄球菌在顽固性CRS的病理生理学中的作用。需要进一步的研究来揭示相关的分子机制。(C)2015 ARS-AAOA有限责任公司
BackgroundChronic rhinosinusitis (CRS) is a cluster of disorders that result in sinonasal mucosal inflammation. Staphylococcus aureus (S. aureus) is associated with severe and recalcitrant CRS. The purpose of our study was to investigate the effect of S. aureus on respiratory epithelial barrier structure and function.MethodsConditioned media from S. aureus reference strains (American Type Culture Collection [ATCC] 13565, 14458, and 25923) was applied to air-liquid interface (ALI) cultures of primary human nasal epithelial cells (HNECs) and transepithelial electrical resistance (TEER) was measured to assess cell-to-cell integrity. Electron microscopy was used to gauge the ciliated area and tight junctions (TJs). Additionally, the expression of the TJ protein zona occludens-1 (ZO-1) was examined via immunofluorescence. Statistical analysis was performed using analysis of variance (ANOVA) with pairwise Bonferroni-adjusted t tests.ResultsSecreted products applied to ALI cultures from S. aureus strain 13565 caused a concentration-dependent decline in electrical impedance compared to controls and reference strains 14458 and 25923 (p < 0.001). Electron microscopy showed a distinct separation between adjacent cells apically, in the region of TJs. The ciliated area was not affected; however, ZO-1 expression became discontinuous in HNECs exposed to the 13565 strain's conditioned media.ConclusionConditioned media of the S. aureus strain 13565 damages the airway epithelium by disrupting the TJs between primary HNECs grown at an ALI. These findings suggest that strain-specific S. aureus-secreted product(s) compromise epithelial barrier function, which may constitute 1 of the roles played by S. aureus in the pathophysiology of recalcitrant CRS. Further research is required to uncover the relevant molecular mechanisms. (C) 2015 ARS-AAOA, LLC