Department of Surgery-Otorhinolaryngology Head and Neck Surgery, The Queen Elizabeth Hospital and the University of Adelaide, Adelaide, SA, Australia.
Int Forum Allergy Rhinol. 2015 Jun;5(6):551-6. doi: 10.1002/alr.21517. Epub 2015 Mar 26.
Chronic 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.
Conditioned 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.
Secreted 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.
Conditioned 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.
慢性鼻鼻窦炎(CRS)是一组导致鼻黏膜炎症的疾病。金黄色葡萄球菌(S. aureus)与严重和难治性 CRS 有关。我们的研究目的是研究金黄色葡萄球菌对呼吸上皮屏障结构和功能的影响。
金黄色葡萄球菌参考株(美国典型培养物保藏中心 [ATCC] 13565、14458 和 25923)的条件培养基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)以评估细胞间完整性。电子显微镜用于评估纤毛面积和紧密连接(TJ)。此外,通过免疫荧光法检测 TJ 蛋白闭合蛋白-1(ZO-1)的表达。使用方差分析(ANOVA)和配对 Bonferroni 调整的 t 检验进行统计分析。
与对照和参考株 14458 和 25923 相比,来自金黄色葡萄球菌菌株 13565 的分泌产物在 ALI 培养物中应用时会导致电阻抗呈浓度依赖性下降(p < 0.001)。电子显微镜显示,在 TJ 区域的细胞顶部分离处相邻细胞之间有明显的分离。纤毛面积不受影响;然而,暴露于 13565 菌株条件培养基的 HNEC 中 ZO-1 的表达变得不连续。
金黄色葡萄球菌菌株 13565 的条件培养基通过破坏在 ALI 上生长的原代 HNEC 之间的 TJ 来破坏气道上皮。这些发现表明,金黄色葡萄球菌分泌的特定菌株产物会损害上皮屏障功能,这可能是金黄色葡萄球菌在难治性 CRS 病理生理学中发挥作用的 1 个角色。需要进一步研究以揭示相关的分子机制。