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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 6

Airway mucus accumulated after elastase administration in mice has a stratified appearance.

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Airway mucus accumulated after elastase administration in mice has a str...
(A) Lower (left) and higher magnification (right) images of an AB/PAS-stained paraffin section showing bronchial walls lined by mucus with lamellar appearance in a mouse exposed to PPE. Representative of 9 animals. Scale bars: left, 200 μm; right, 50 μm. In this and the other pictures, the white line marks the mucus layer. (B) A paraffin section from a mouse exposed to PPE stained with specific antibodies against Muc5b (green) and Muc5ac (red), in addition to nuclear stain (blue), shows that this stratified structure consisted Muc5b and Muc5ac. Representative of 3 animals. Scale bar: 10 μm. (C) The stratified appearance of mucus has been observed in the distal colon, where the mucus is organized to separate the epithelium from bacterial contact in humans as well as mice. Representative image showing mouse distal colon in an AB/PAS-stained paraffin section. Scale bar: 50 μm. In airway sections from patients with (D) COPD and (E) cystic fibrosis stained with AB/PAS, stratified mucus can be observed covering the epithelium. Scale bars: 50 μm. In mice as well as humans, mucus does not accumulate in the airways in (F) control subjects. Section from patients with (G) COPD and (H) cystic fibrosis stained with antibodies against MUC5B (green) and MUC5AC (red), in addition to nuclear stain (blue), shows a stratified structure of MUC5B and MUC5AC in the mucus. The MUC5B cell surface staining in F could be due to cross-reactivity. Scale bars: 20 μm.

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