Chowdhury Ferdousi, Howat William J, Phillips Gary J, Lackie Peter M
Allergy and Inflammation Research, Infection Inflammation and Immunity Division, School of Medicine, University of Southampton, Southampton, UK.
Exp Lung Res. 2010 Feb;36(1):1-11. doi: 10.3109/01902140903026582.
Environmental particulates impact first on airway epithelium, whereas circulating infiltrating cells are recruited through the underlying endothelium. An effective cellular immune response requires coordination between endothelium and epithelium. The authors have developed a bilayer culture model consisting of human bronchial epithelial derived cells (16HBE 14o-) and human umbilical vein endothelial cells (HUVECs) cultured as confluent layers on either side of a porous membrane. Confocal microscopy with epithelial and endothelial-specific antibodies showed segregated cell layers. By scanning and transmission electron microscopy, both cell types are polarized and tight junctions formed at the apical interface between cells. Epithelial cells grown in a bilayer showed significantly increased transepithelial resistance (TER) of 2260 +/- 64 Omega.cm(2) compared to epithelial or endothelial monolayers alone (1400 +/- 70 or 80 +/- 12 Omega.cm(2), respectively). This reflected decreased permeability and was unrelated to cell density or height. Increased TER coincided with increased occludin mRNA and protein in the epithelial cell layer as determined by polymerase chain reaction (PCR) and immunoblotting. Conditioned medium showed that decreased permeability was mediated by soluble endothelial-derived factor(s). This model reflects the in vivo relationship of human airway endothelial cells and epithelial cells. Altered tight junction permeability in cocultures indicates that these cells can work together as an active part of the mucosal barrier.
环境颗粒物首先影响气道上皮，而循环浸润细胞则通过下层内皮细胞募集。有效的细胞免疫反应需要内皮细胞和上皮细胞之间的协调。作者开发了一种双层培养模型，该模型由人支气管上皮衍生细胞（16HBE 14o-）和人脐静脉内皮细胞（HUVECs）组成，在多孔膜的两侧以汇合层的形式培养。用上皮细胞和内皮细胞特异性抗体进行的共聚焦显微镜检查显示细胞层分离。通过扫描电子显微镜和透射电子显微镜观察，两种细胞类型均呈极化状态，且在细胞顶端界面形成紧密连接。与单独的上皮细胞或内皮细胞单层（分别为1400 +/- 70或80 +/- 12 Ω.cm2）相比，在双层中生长的上皮细胞的跨上皮电阻（TER）显著增加，达到2260 +/- 64 Ω.cm2。这反映了通透性降低，且与细胞密度或高度无关。通过聚合酶链反应（PCR）和免疫印迹法测定，TER增加与上皮细胞层中闭合蛋白mRNA和蛋白质的增加一致。条件培养基显示，通透性降低是由可溶性内皮衍生因子介导的。该模型反映了人气道内皮细胞和上皮细胞的体内关系。共培养中紧密连接通透性的改变表明，这些细胞可以作为黏膜屏障的一个活跃部分共同发挥作用。
