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In Vitro Spatial and Temporal Analysis of Mycoplasma pneumoniae Colonization of Human Airway Epithelium

机译：人气道上皮细胞肺炎支原体定植的体外时空分析

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Mycoplasma pneumoniae is an important cause of respiratory disease, especially in school-age children and young adults. We employed normal human bronchial epithelial (NHBE) cells in air-liquid interface culture to study the interaction of M. pneumoniae with differentiated airway epithelium. These airway cells, when grown in air-liquid interface culture, polarize, form tight junctions, produce mucus, and develop ciliary function. We examined both qualitatively and quantitatively the role of mycoplasma gliding motility in the colonization pattern of developing airway cells, comparing wild-type M. pneumoniae and mutants thereof with moderate to severe defects in gliding motility. Adherence assays with radiolabeled mycoplasmas demonstrated a dramatic reduction in binding for all strains with airway cell polarization, independent of acquisition of mucociliary function. Adherence levels dropped further once NHBE cells achieved terminal differentiation, with mucociliary activity strongly selecting for full gliding competence. Analysis over time by confocal microscopy demonstrated a distinct colonization pattern that appeared to originate primarily with ciliated cells, but lateral spread from the base of the cilia was slower than expected. The data support a model in which the mucociliary apparatus impairs colonization yet cilia provide a conduit for mycoplasma access to the host cell surface and suggest acquisition of a barrier function, perhaps associated with tethered mucin levels, with NHBE cell polarization.
机译：肺炎支原体是呼吸系统疾病的重要原因，尤其是在学龄儿童和年轻人中。我们在气液界面培养中采用正常人支气管上皮细胞（NHBE）来研究肺炎支原体与分化气道上皮的相互作用。这些气道细胞在气液界面培养物中生长时，会极化，形成紧密的连接，产生粘液并发展睫状功能。我们定性和定量地检查了支原体滑行运动在发育气道细胞定植模式中的作用，比较了野生型肺炎支原体及其突变体在滑行运动中具有中度至重度缺陷。放射性标记的支原体的粘附试验表明，与气道细胞极化有关的所有菌株的结合均显着降低，而与粘液纤毛功能的获得无关。 NHBE细胞达到终末分化后，黏附水平进一步下降，粘膜纤毛活性强烈地选择了完整的滑行能力。通过共聚焦显微镜对时间的分析显示出明显的定居模式，该定居模式似乎主要起源于纤毛细胞，但是从纤毛底部的侧向扩散比预期的要慢。数据支持一种模型，其中粘膜纤毛装置损害定植，而纤毛为支原体进入宿主细胞表面提供了导管，并暗示了与NHBE细胞极化相关的屏障功能的获得，可能与栓系粘蛋白水平有关。

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