Epithelial Pathobiology and Mucosal Inflammation Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA 30308, USA.
Int Forum Allergy Rhinol. 2013 Jan;3(1):19-25. doi: 10.1002/alr.21077. Epub 2012 Aug 27.
Chronic rhinosinusitis (CRS) is an inflammatory upper-airway disease with numerous etiologies. Patients with a characteristic subtype of CRS, allergic fungal rhinosinusitis (AFRS), display increased expression of T helper 2 (Th2) cytokines and antigen-specific immunoglobulin E (IgE). Various sinonasal inflammatory conditions are associated with alterations in epithelial barrier function. The aim of this study was to compare epithelial permeability and intercellular junctional protein expression among cultured primary sinonasal cells from AFRS patients vs noninflammatory controls.
Epithelial cells isolated from paranasal sinus mucosa of AFRS and noninflammatory control patients were grown to confluence on permeable supports and transitioned to air-liquid interface (ALI). Transepithelial resistance (TER) was measured with a horizontal Ussing chamber to characterize the functional permeability of each cell type. After TER recordings were complete, a panel of intercellular junctional proteins was assessed by Western blot and immunofluorescence labeling followed by confocal microscopy.
After 12 samples were measured from each group, we observed a 41% mean decrease in TER in AFRS cells (296 ± 89 ohms × cm(2) ) compared to control (503 ± 134 ohms × cm(2) , p = 0.006). TER deficits observed in AFRS were associated with decreased expression of the tight junction proteins occludin and junctional adhesion molecule-A (JAM-A), and increased expression of a leaky tight junction protein claudin-2.
Cultured sinonasal epithelium from AFRS patients displayed increased epithelial permeability and altered expression of intercellular junctional proteins. Given that these cells were not incubated with inflammatory cytokines in vitro, the cultured AFRS epithelial alterations may represent a retained modification in protein expression from the in vivo phenotype.
慢性鼻-鼻窦炎（CRS）是一种具有多种病因的炎症性上呼吸道疾病。具有特征性亚类的 CRS（变应性真菌性鼻-鼻窦炎，AFRS）的患者表现出 Th2 细胞因子和抗原特异性免疫球蛋白 E（IgE）表达增加。各种鼻-鼻窦炎症性疾病与上皮屏障功能改变有关。本研究旨在比较 AFRS 患者与非炎症性对照患者的培养原代鼻-鼻窦细胞的上皮通透性和细胞间连接蛋白表达。
从 AFRS 和非炎症性对照患者的鼻窦黏膜中分离出上皮细胞，在可渗透的支持物上生长至汇合，然后过渡到气-液界面（ALI）。用水平 Ussing 室测量跨上皮电阻（TER）以表征每种细胞类型的功能通透性。在完成 TER 记录后，通过 Western blot 和免疫荧光标记以及共聚焦显微镜评估细胞间连接蛋白的表达。
从每组测量了 12 个样本后，我们观察到 AFRS 细胞的 TER 平均下降了 41%（296 ± 89 欧姆×厘米²），而对照组为 503 ± 134 欧姆×厘米²（p = 0.006）。在 AFRS 中观察到的 TER 缺陷与紧密连接蛋白闭合蛋白和连接黏附分子-A（JAM-A）的表达减少以及渗漏性紧密连接蛋白 Claudin-2 的表达增加有关。
来自 AFRS 患者的培养鼻-鼻窦上皮显示出增加的上皮通透性和细胞间连接蛋白表达的改变。鉴于这些细胞在体外未用炎症细胞因子孵育，培养的 AFRS 上皮改变可能代表体内表型保留的蛋白表达改变。
