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Analysis of pathological changes in the epithelium in COVID-19 patient airways

Wenguang Yin, Weitao Cao, Guangde Zhou, Lifei Wang, Jing Sun, Airu Zhu, Zhongfang Wang, Yumin Zhou, Xiaoqing Liu, Yimin Li, Nanshan Zhong, Jincun Zhao, Lei Liu, Pixin Ran
ERJ Open Research 2021 7: 00690-2020; DOI: 10.1183/23120541.00690-2020
Wenguang Yin
1State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
4These authors contributed equally
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Weitao Cao
1State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
4These authors contributed equally
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Guangde Zhou
2Shenzhen Key Laboratory of Pathogen and Immunity, National Clinical Research Center for Infectious Disease, State Key Discipline of Infectious Disease, Shenzhen Third People's Hospital, Second Hospital Affiliated to Southern University of Science and Technology, Shenzhen, China
4These authors contributed equally
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Lifei Wang
3Dept of Radiology, National Clinical Research Center for Infectious Disease, Third People's Hospital, Second Hospital Affiliated to Southern University of Science and Technology, Shenzhen, China
4These authors contributed equally
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Jing Sun
1State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
4These authors contributed equally
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Airu Zhu
1State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
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Zhongfang Wang
1State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
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Yumin Zhou
1State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
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Xiaoqing Liu
1State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
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Yimin Li
1State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
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Nanshan Zhong
1State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
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Jincun Zhao
1State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
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Lei Liu
2Shenzhen Key Laboratory of Pathogen and Immunity, National Clinical Research Center for Infectious Disease, State Key Discipline of Infectious Disease, Shenzhen Third People's Hospital, Second Hospital Affiliated to Southern University of Science and Technology, Shenzhen, China
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Pixin Ran
1State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
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  • ORCID record for Pixin Ran
  • For correspondence: pxran@gzhmu.edu.cn
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    FIGURE 1

    Coronavirus disease 2019 (COVID-19) patient lungs exhibit mucus plugs and pathological changes in the airway epithelium. Chest radiographs obtained on a) 12 January 2020, b) 17 January 2020 and c) 22 January 2020. d, e) Representative lung CT image on 13 January 2020. f, g) Representative transverse lung CT image on 23 January 2020. Chest radiographs obtained on h) 30 January 2020 i) 1 February 2020 and j) 15 February 2020. k) Representative images of lung tissue sections from the COVID-19 patient stained with haematoxylin and eosin (HE). Asterisks indicate airways. l) Representative images of lung tissue sections from the COVID-19 patient stained with periodic acid–Schiff (PAS). The arrows point to goblet cells. Asterisks indicate mucus plugs in the airways. m) Immunostaining for MUC5AC (green) and DAPI staining (blue) of sections of larger distal bronchioles from healthy controls (n=3) and the COVID-19 patient. The arrow points to the mucus plug. The arrowhead points to the goblet cell. n) Quantification of the number of larger distal bronchioles plugged by mucus in healthy controls (n=3) and the COVID-19 patient. o) Quantification of the number of larger distal bronchioles with MUC5AC+ cells in healthy controls (n=3) and the COVID-19 patient. p) Immunostaining for MUC5AC (green) and DAPI staining (blue) of sections of the smaller distal bronchioles from healthy controls (n=3) and the COVID-19 patient. The arrow points to the mucus plug. The arrowhead points to the goblet cell. q) Quantification of the number of smaller distal bronchioles plugged by mucus in healthy controls (n=3) and the COVID-19 patient. r) Quantification of the number of smaller distal bronchioles with MCU5AC+ cells in healthy controls (n=3) and the COVID-19 patient. s) Relative MUC5AC mRNA levels in the lungs from healthy controls and the COVID-19 patient. t) Immunostaining for MUC5B (red) and DAPI staining (blue) of sections of the airways from healthy controls (n=3) and the COVID-19 patient. u) Quantification of mean fluorescence intensity of MUC5B immunostaining in the airways in healthy controls (n=3) and the COVID-19 patient. v) Relative MUC5B mRNA levels in the lungs from healthy controls and the COVID-19 patient. w) Immunostaining for acetylated α-tubulin (green) and DAPI staining (blue) of sections of small bronchioles from healthy controls (n=3) and the COVID-19 patient. x) Quantification of the relative number of acetylated α-tubulin+ multiciliated cells in small bronchioles in healthy controls (n=3) and the COVID-19 patient. y) Immunostaining for CC16 (red) and DAPI staining (blue) of sections of distal bronchioles from healthy controls (n=3) and the COVID-19 patient. z) Quantification of the relative number of CC16+ cells in distal bronchioles in healthy controls (n=3) and the COVID-19 patient. aa) Relative CC16 mRNA levels in the lungs from healthy controls and the COVID-19 patient. bb) Quantification of CC16 levels in serum from healthy controls (n=7) and severe and critical COVID-19 patients (n=9). cc) Immunostaining for interleukin (IL)-13 (red) and DAPI staining (blue) of sections of the lungs from healthy controls (n=3) and the COVID-19 patient. dd) Quantification of mean fluorescence intensity of IL-13 immunostaining in the lungs in healthy controls (n=3) and the COVID-19 patient. ee) Representative images of cross sections of air–liquid interface (ALI) cultures stained with HE. The arrow points to cilia. ff) Trans-epithelial electrical resistance (TEER) measurements. gg) Immunostaining for MUC5AC (red), severe acute respiratory syndrome coronavirus 2 nucleocapsid proteins (SARS-CoV-2 N; green) and DAPI staining (blue) in human bronchial epithelial cells at the ALI after a 14-day differentiation. Scale bars: 100 μm (k, l, m, p, gg), 20 μm (t, w, y, cc, ee). *: p<0.05; **: p<0.01; ***: p<0.001; ****: p <0.0001; unpaired t-test, mean±sd.

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Vol 7 Issue 2 Table of Contents
ERJ Open Research: 7 (2)
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Analysis of pathological changes in the epithelium in COVID-19 patient airways
Wenguang Yin, Weitao Cao, Guangde Zhou, Lifei Wang, Jing Sun, Airu Zhu, Zhongfang Wang, Yumin Zhou, Xiaoqing Liu, Yimin Li, Nanshan Zhong, Jincun Zhao, Lei Liu, Pixin Ran
ERJ Open Research Apr 2021, 7 (2) 00690-2020; DOI: 10.1183/23120541.00690-2020

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Analysis of pathological changes in the epithelium in COVID-19 patient airways
Wenguang Yin, Weitao Cao, Guangde Zhou, Lifei Wang, Jing Sun, Airu Zhu, Zhongfang Wang, Yumin Zhou, Xiaoqing Liu, Yimin Li, Nanshan Zhong, Jincun Zhao, Lei Liu, Pixin Ran
ERJ Open Research Apr 2021, 7 (2) 00690-2020; DOI: 10.1183/23120541.00690-2020
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