Pseudomonas aeruginosa-Mediated Damage Requires Distinct Receptors at the Apical and Basolateral Surfaces of the Polarized Epithelium
Pseudomonas aeruginosa-Mediated Damage Requires Distinct Receptors at the Apical and Basolateral Surfaces of the Polarized Epithelium
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DOI：
10.1128/iai.01215-09
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发表时间：
2010-03-01
影响因子：
3.1
通讯作者：
Engel, Joanne N.
中科院分区：
文献类型：
--
作者：
Bucior, Iwona;Mostov, Keith;Engel, Joanne N.
铜绿假单胞菌是一种重要的人类机会致病菌，它利用上皮损伤来建立感染。我们已经严格探索了N-糖蛋白和硫酸乙酰肝素蛋白聚糖（HSPG）在铜绿假单胞菌介导的附着和随后的极化上皮的顶端（AP）和基底外侧（BL）表面的下游事件中的作用。我们证明，在AP表面的N-聚糖链是必要的和足够的结合，入侵和细胞毒性的肾（MDCK）和气道（Calu-3）细胞生长在各种状态的极化Transwell过滤器。上调N-糖基化增强结合，而药理学抑制N-糖基化或感染N-糖基化缺陷的MDCK细胞导致结合降低。相反，在BL表面，HSPG的HS部分介导铜绿假单胞菌结合、细胞毒性和侵袭。在不完全极化的上皮中，AP表面的HSPG丰度增加，解释了其对铜绿假单胞菌定植和损伤的易感性增加。使用MDCK细胞生长为三维囊肿作为上皮器官的模型，我们表明，铜绿假单胞菌特异性共定位与HS丰富的地区在BL膜，但复杂的N-聚糖在AP表面。最后，铜绿假单胞菌结合到HS链和塑料表面上包被的N-聚糖，显示出对分离的HS链的最高结合亲和力。总之，这些发现表明铜绿假单胞菌识别极化上皮的AP和BL表面上的不同受体。N-聚糖链组成和/或HSPG分布的变化可以解释受损上皮对铜绿假单胞菌的敏感性增强。
Pseudomonas aeruginosa, an important opportunistic pathogen of humans, exploits epithelial damage to establish infection. We have rigorously explored the role of N-glycoproteins and heparan sulfate proteoglycans (HSPGs) in P. aeruginosa-mediated attachment and subsequent downstream events at the apical (AP) and basolateral (BL) surfaces of polarized epithelium. We demonstrate that the N-glycan chains at the AP surface are necessary and sufficient for binding, invasion, and cytotoxicity to kidney (MDCK) and airway (Calu-3) cells grown at various states of polarization on Transwell filters. Upregulation of N-glycosylation enhanced binding, whereas pharmacologic inhibition of N-glycosylation or infection of MDCK cells defective in N-glycosylation resulted in decreased binding. In contrast, at the BL surface, the HS moiety of HSPGs mediated P. aeruginosa binding, cytotoxicity, and invasion. In incompletely polarized epithelium, HSPG abundance was increased at the AP surface, explaining its increased susceptibility to P. aeruginosa colonization and damage. Using MDCK cells grown as three-dimensional cysts as a model for epithelial organs, we show that P. aeruginosa specifically colocalized with HS-rich areas at the BL membrane but with complex N-glycans at the AP surface. Finally, P. aeruginosa bound to HS chains and N-glycans coated on plastic surfaces, showing the highest binding affinity toward isolated HS chains. Together, these findings demonstrate that P. aeruginosa recognizes distinct receptors on the AP and BL surfaces of polarized epithelium. Changes in the composition of N-glycan chains and/or in the distribution of HSPGs may explain the enhanced susceptibility of damaged epithelium to P. aeruginosa.