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Attached stratified mucus separates bacteria from the epithelial cells in COPD lungs
Joan Antoni Fernández-Blanco, Dalia Fakih, Liisa Arike, Ana M. Rodríguez-Piñeiro, Beatriz Martínez-Abad, Elin Skansebo, Sonya Jackson, James Root, Dave Singh, Christopher McCrae, Christopher M. Evans, Annika Åstrand, Anna Ermund, Gunnar C. Hansson
Joan Antoni Fernández-Blanco, Dalia Fakih, Liisa Arike, Ana M. Rodríguez-Piñeiro, Beatriz Martínez-Abad, Elin Skansebo, Sonya Jackson, James Root, Dave Singh, Christopher McCrae, Christopher M. Evans, Annika Åstrand, Anna Ermund, Gunnar C. Hansson
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Research Article Pulmonology

Attached stratified mucus separates bacteria from the epithelial cells in COPD lungs

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Abstract

The respiratory tract is normally kept essentially free of bacteria by cilia-mediated mucus transport, but in chronic obstructive pulmonary disease (COPD) and cystic fibrosis (CF), bacteria and mucus accumulates instead. To address the mechanisms behind the mucus accumulation, the proteome of bronchoalveolar lavages from COPD patients and mucus collected in an elastase-induced mouse model of COPD was analyzed, revealing similarities with each other and with the protein content in colonic mucus. Moreover, stratified laminated sheets of mucus were observed in airways from patients with CF and COPD and in elastase-exposed mice. On the other hand, the mucus accumulation in the elastase model was reduced in Muc5b-KO mice. While mucus plugs were removed from airways by washing with hypertonic saline in the elastase model, mucus remained adherent to epithelial cells. Bacteria were trapped on this mucus, whereas, in non–elastase-treated mice, bacteria were found on the epithelial cells. We propose that the adherence of mucus to epithelial cells observed in CF, COPD, and the elastase-induced mouse model of COPD separates bacteria from the surface cells and, thus, protects the respiratory epithelium.

Authors

Joan Antoni Fernández-Blanco, Dalia Fakih, Liisa Arike, Ana M. Rodríguez-Piñeiro, Beatriz Martínez-Abad, Elin Skansebo, Sonya Jackson, James Root, Dave Singh, Christopher McCrae, Christopher M. Evans, Annika Åstrand, Anna Ermund, Gunnar C. Hansson

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Figure 5

Airway epithelial cell proteome changes after elastase (PPE) administration.

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Airway epithelial cell proteome changes after elastase (PPE) administrat...
(A) H&E-stained section of bronchi before (left) and after epithelial cell isolation (right). Scale bars: (left) 200 μm, (right) 50 μm. (B) Heatmap of the most changed proteins in the airway epithelial cell proteome after PPE exposure. (C and D) The most abundant proteins in airway epithelial cells detected by label-free proteomics after saline (C) and PPE (D) instillation. (E) Label-free quantification (LFQ) of the most changed proteins after PPE exposure, n = 7 pools/group with 3 mice/pool, data presented as median ± IQR, *P = 0.017, **P = 0.006, ***P = 0.0006, Mann-Whitney U test. The detection limit for Muc5ac was set to 1 × 106. (F) Absolute quantification of Muc5ac and Muc5b with isotopically labeled peptides by mass spectrometry; n = 8 pools/group with 3 mice/pool, data presented as median ± IQR, **P = 0.001, Mann-Whitney U test. Detection limit for Muc5ac was set to 1 fmol. (G) Ratio of Muc5ac to Muc5b in isolated mucus plugs, BALF supernatants (BALF SN), and epithelial cells, n = 6–12 samples/group. Data presented as median ± IQR, *P = 0.04, **P = 0.0016, Kruskal-Wallis and Dunn’s multiple comparisons test. Chi3l4, chitinase-3-like protein 4; Clca1, chloride channel accessory 1; Retnla, resistin-like α; Chi3l3, chitinase-3-like protein 3; Ltf, lactotransferrin; C3, complement C3; Agr2, anterior gradient protein 2 homolog; Cp, ceruloplasmin; Bpifb1, BPI fold-containing family B member 1; Chi3l1, chitinase-3-like protein 1; Scin, adseverin; Fcgbp, Fcgbp protein; Pla2g4c, Pla2g4c protein; St3gal4, β-galactoside α-2,3-sialyltransferase 4; Tff2, trefoil factor 2; Pglyrp1, peptidoglycan recognition protein 1; Reg3g, regenerating islet-derived protein 3-γ; Qsox1, sulfhydryl oxidase 1; Chia, acidic mammalian chitinase; Chad, chondroadherin; Fer1l6, Fer-1-like 6; Fn1, fibronectin; Cbr2, carbonyl reductase [NADPH] 2; Aldh1a1, retinal dehydrogenase 1; Sec14l3, SEC14-like 3; Hist1h2bj, histone H2B; Scgb1a1, uteroglobin.

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