Department of Medicine, University of Maryland School of Medicine, Baltimore, MD, USA.
U.S. Department of Veterans Affairs, Baltimore VA Medical Center, Baltimore, MD, USA.
Sci Rep. 2021 Nov 22;11(1):22725. doi: 10.1038/s41598-021-02242-x.
We previously reported that flagellin-expressing Pseudomonas aeruginosa (Pa) provokes NEU1 sialidase-mediated MUC1 ectodomain (MUC1-ED) desialylation and MUC1-ED shedding from murine lungs in vivo. Here, we asked whether Pa in the lungs of patients with ventilator-associated pneumonia might also increase MUC1-ED shedding. The levels of MUC1-ED and Pa-expressed flagellin were dramatically elevated in bronchoalveolar lavage fluid (BALF) harvested from Pa-infected patients, and each flagellin level, in turn, predicted MUC1-ED shedding in the same patient. Desialylated MUC1-ED was only detected in BALF of Pa-infected patients. Clinical Pa strains increased MUC1-ED shedding from cultured human alveolar epithelia, and FlaA and FlaB flagellin-expressing strains provoked comparable levels of MUC1-ED shedding. A flagellin-deficient isogenic mutant generated dramatically reduced MUC1-ED shedding compared with the flagellin-expressing wild-type strain, and purified FlaA and FlaB recapitulated the effect of intact bacteria. Pa:MUC1-ED complexes were detected in the supernatants of alveolar epithelia exposed to wild-type Pa, but not to the flagellin-deficient Pa strain. Finally, human recombinant MUC1-ED dose-dependently disrupted multiple flagellin-driven processes, including Pa motility, Pa biofilm formation, and Pa adhesion to human alveolar epithelia, while enhancing human neutrophil-mediated Pa phagocytosis. Therefore, shed desialylated MUC1-ED functions as a novel flagellin-targeting, Pa-responsive decoy receptor that participates in the host response to Pa at the airway epithelial surface.
我们之前曾报道过,表达鞭毛蛋白的铜绿假单胞菌(Pa)会引发 NEU1 唾液酸酶介导的 MUC1 外显子(MUC1-ED)去唾液酸化和 MUC1-ED 从体内的小鼠肺部脱落。在这里,我们想知道呼吸机相关性肺炎患者肺部的 Pa 是否也会增加 MUC1-ED 脱落。从 Pa 感染患者的支气管肺泡灌洗液(BALF)中,MUC1-ED 和 Pa 表达的鞭毛蛋白水平显著升高,并且每个鞭毛蛋白水平反过来又预测了同一患者的 MUC1-ED 脱落。仅在 Pa 感染患者的 BALF 中检测到去唾液酸化的 MUC1-ED。临床 Pa 菌株增加了培养的人肺泡上皮细胞中 MUC1-ED 的脱落,并且 FlaA 和 FlaB 表达鞭毛蛋白的菌株引发了相当水平的 MUC1-ED 脱落。与表达鞭毛蛋白的野生型菌株相比,鞭毛蛋白缺陷的同基因突变体产生的 MUC1-ED 脱落明显减少,而纯化的 FlaA 和 FlaB 则重现了完整细菌的作用。在暴露于野生型 Pa 的肺泡上皮细胞的上清液中检测到 Pa:MUC1-ED 复合物,但在鞭毛蛋白缺陷的 Pa 菌株中没有检测到。最后,人重组 MUC1-ED 剂量依赖性地破坏了多个鞭毛蛋白驱动的过程,包括 Pa 运动性、Pa 生物膜形成和 Pa 与人类肺泡上皮的黏附,同时增强了人中性粒细胞介导的 Pa 吞噬作用。因此,脱落的去唾液酸化的 MUC1-ED 作为一种新型的鞭毛蛋白靶向、Pa 反应性诱饵受体发挥作用,参与了气道上皮表面宿主对 Pa 的反应。