Flagellin shifts 3D bronchospheres towards mucus hyperproduction
Abstract Cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD) are associated with acute and chronic bacterial infections of the lung. Excessive differentiation of basal cells to mucus-producing goblet cells can result in mucus hyperproduction and loss of mucociliary clearance in the...
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doaj-c720dc3eef364c3e911034fc54a8df132020-11-25T03:49:27ZengBMCRespiratory Research1465-993X2020-08-012111610.1186/s12931-020-01486-xFlagellin shifts 3D bronchospheres towards mucus hyperproductionRichard F. Sprott0Felix Ritzmann1Frank Langer2Yiwen Yao3Christian Herr4Yvonne Kohl5Thomas Tschernig6Robert Bals7Christoph Beisswenger8Department of Internal Medicine V – Pulmonology, Allergology and Respiratory Critical Care Medicine, Saarland UniversityDepartment of Internal Medicine V – Pulmonology, Allergology and Respiratory Critical Care Medicine, Saarland UniversityDepartment of Thoracic and Cardiovascular Surgery, Saarland UniversityDepartment of Internal Medicine V – Pulmonology, Allergology and Respiratory Critical Care Medicine, Saarland UniversityDepartment of Internal Medicine V – Pulmonology, Allergology and Respiratory Critical Care Medicine, Saarland UniversityDepartment Bioprocessing & Bioanalytics, Fraunhofer Institute for Biomedical EngineeringInstitute of Anatomy and Cell Biology, Saarland UniversityDepartment of Internal Medicine V – Pulmonology, Allergology and Respiratory Critical Care Medicine, Saarland UniversityDepartment of Internal Medicine V – Pulmonology, Allergology and Respiratory Critical Care Medicine, Saarland UniversityAbstract Cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD) are associated with acute and chronic bacterial infections of the lung. Excessive differentiation of basal cells to mucus-producing goblet cells can result in mucus hyperproduction and loss of mucociliary clearance in the airways of CF and COPD patients. Here, we aimed to investigate the effect of pathogen-associated molecular patterns (PAMPs) on the differentiation of human 3D bronchospheres. Primary human bronchial epithelial cells (HBECs) were differentiated to bronchospheres in the presence of bacterial flagellin and LPS and the synthetic Toll-like receptor (TLR) ligands Pam3CSK4 (TLR-2) and polyinosinic:polycytidylic acid (pIC, TLR-3). Electron and fluorescence microscopy showed that the differentiation of bronchospheres associated with the formation of lumina and appearance of cilia within 30 days after seeding. Incubation with flagellin resulted in a decreased formation of lumina and loss of cilia formation. Incubation with Pam3CSK, pIC, and LPS did not significantly affect formation of lumina and ciliation. Mucus production was strongly increased in response to flagellin and, to a lesser degree, in response to Pam3CSK4. Our results indicate that bacterial factors, such as flagellin, drive the differentiation of the respiratory epithelium towards mucus hyperproduction.http://link.springer.com/article/10.1186/s12931-020-01486-xOrganoidsBronchospheresMucusPAMPsFlagellin |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Richard F. Sprott Felix Ritzmann Frank Langer Yiwen Yao Christian Herr Yvonne Kohl Thomas Tschernig Robert Bals Christoph Beisswenger |
spellingShingle |
Richard F. Sprott Felix Ritzmann Frank Langer Yiwen Yao Christian Herr Yvonne Kohl Thomas Tschernig Robert Bals Christoph Beisswenger Flagellin shifts 3D bronchospheres towards mucus hyperproduction Respiratory Research Organoids Bronchospheres Mucus PAMPs Flagellin |
author_facet |
Richard F. Sprott Felix Ritzmann Frank Langer Yiwen Yao Christian Herr Yvonne Kohl Thomas Tschernig Robert Bals Christoph Beisswenger |
author_sort |
Richard F. Sprott |
title |
Flagellin shifts 3D bronchospheres towards mucus hyperproduction |
title_short |
Flagellin shifts 3D bronchospheres towards mucus hyperproduction |
title_full |
Flagellin shifts 3D bronchospheres towards mucus hyperproduction |
title_fullStr |
Flagellin shifts 3D bronchospheres towards mucus hyperproduction |
title_full_unstemmed |
Flagellin shifts 3D bronchospheres towards mucus hyperproduction |
title_sort |
flagellin shifts 3d bronchospheres towards mucus hyperproduction |
publisher |
BMC |
series |
Respiratory Research |
issn |
1465-993X |
publishDate |
2020-08-01 |
description |
Abstract Cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD) are associated with acute and chronic bacterial infections of the lung. Excessive differentiation of basal cells to mucus-producing goblet cells can result in mucus hyperproduction and loss of mucociliary clearance in the airways of CF and COPD patients. Here, we aimed to investigate the effect of pathogen-associated molecular patterns (PAMPs) on the differentiation of human 3D bronchospheres. Primary human bronchial epithelial cells (HBECs) were differentiated to bronchospheres in the presence of bacterial flagellin and LPS and the synthetic Toll-like receptor (TLR) ligands Pam3CSK4 (TLR-2) and polyinosinic:polycytidylic acid (pIC, TLR-3). Electron and fluorescence microscopy showed that the differentiation of bronchospheres associated with the formation of lumina and appearance of cilia within 30 days after seeding. Incubation with flagellin resulted in a decreased formation of lumina and loss of cilia formation. Incubation with Pam3CSK, pIC, and LPS did not significantly affect formation of lumina and ciliation. Mucus production was strongly increased in response to flagellin and, to a lesser degree, in response to Pam3CSK4. Our results indicate that bacterial factors, such as flagellin, drive the differentiation of the respiratory epithelium towards mucus hyperproduction. |
topic |
Organoids Bronchospheres Mucus PAMPs Flagellin |
url |
http://link.springer.com/article/10.1186/s12931-020-01486-x |
work_keys_str_mv |
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