Pseudomonas aeruginosa internalization by human epithelial respiratory cells depends on cell differentiation, polarity, and junctional complex integrity
Pseudomonas aeruginosa internalization by human epithelial respiratory cells depends on cell differentiation, polarity, and junctional complex integrity
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DOI：
10.1165/ajrcmb.20.5.3408
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发表时间：
1999-05-01
影响因子：
6.4
通讯作者：
Puchelle, E
中科院分区：
文献类型：
--
作者：
Plotkowski, MC;de Bentzmann, S;Puchelle, E
上皮呼吸细胞系对铜绿假单胞菌的内化依赖于囊性纤维化跨膜电导调节蛋白（CFTR）。因为我们仅在不表达顶端CFTR的细胞中观察到细胞内（IC）铜绿假单胞菌，所以我们解决了上皮细胞的细菌内化是否取决于细胞分化程度和极性的问题。通过庆大霉素排除测定评估原代培养物中的人上皮呼吸细胞以及在厚胶原凝胶或薄胶原膜上培养的16个人支气管上皮140细胞系对毛毛P、铜绿假单胞菌PAO-1和PAK的内化。在厚凝胶上培养的细胞已分化、极化且紧密。它们在顶膜上表现出 CFTR，在基底膜上表达 β1 整合素，排除硝酸镧，并均匀表达 ZO-1 蛋白。相反，在薄膜上培养的细胞中，CFTR 主要存在于细胞质中，而 β1 整合素则在上膜上检测到。大多数在薄膜上培养的细胞不排除硝酸镧，并且很少表达ZO-1蛋白。在厚胶原基质和薄胶原基质上生长的细胞在细菌内化方面存在显着差异：在凝胶上培养的细胞中无法检测到IC细菌，而从薄膜培养的细胞中分离出高IC细菌浓度。用乙二胺四乙酸处理薄膜上培养的细胞，进一步破坏细胞间连接，显着增强铜绿假单胞菌的内化。我们的结果表明，上皮呼吸细胞对铜绿假单胞菌的内化并不依赖于上皮细胞表面的 CFTR 蛋白表达，而是依赖于细胞极性和连接复合体的完整性。
Internalization of Pseudomonas aeruginosa by epithelial respiratory cell lines has been suggested to be dependent on the cystic fibrosis transmembrane conductance regulator (CFTR) protein. Because we have observed intracellular (IC) P. aeruginosa only in cells that do not express apical CFTR, we addressed the question of whether bacterial internalization by epithelial cells depends on the degree of cell differentiation and polarity. Internalization of piliated P, aeruginosa PAO-1 and PAK by human epithelial respiratory cells in primary culture and by the 16 human bronchial epithelial 140- cell line cultured either on thick collagen gels or on thin collagen films was evaluated by the gentamicin exclusion assay. Cells cultured on thick gels were differentiated, polarized, and tight. They exhibited CFTR at their apical membranes, expressed beta 1 integrins at their basal membranes, excluded lanthanum nitrate, and uniformly expressed ZO-1 protein. In contrast, in cells cultured on thin films, CFTR was present mainly in the cytoplasm, whereas beta 1 integrins were detected at epical membranes. Most cells cultured on thin films did not exclude lanthanum nitrate and rarely expressed ZO-1 protein. Cells grown on thick and thin collagen substrates differed markedly in bacterial internalization: no IC bacteria could be detected in cells cultured on gels, whereas high IC bacterial concentrations were isolated from cells cultured on thin films. Treatment of cells cultured on thin films with ethylenediaminetetraacetic acid, to disrupt intercellular junctions further, significantly enhanced P. aeruginosa internalization. Our results suggest that P. aeruginosa internalization by epithelial respiratory cells does not depend on CFTR protein expression at the epithelial cell surface but rather on cell polarity and junctional complex integrity.