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Activity of Tracheal Cytotoxin of Bordetella pertussis in a Human Tracheobronchial 3D Tissue Model
David K Kessie1, Nina Lodes2, Heike Oberwinkler2
1Biocentre, Chair of Microbiology, University of Würzburg, Würzburg, Germany.
Abstract:
Bordetella pertussis is a highly contagious pathogen which causes whooping cough in humans. A major pathophysiology of infection is the extrusion of ciliated cells and subsequent disruption of the respiratory mucosa. Tracheal cytotoxin (TCT) is the only virulence factor produced by B. pertussis that has been able to recapitulate this pathology in animal models. This pathophysiology is well characterized in a hamster tracheal model, but human data are lacking due to scarcity of donor material. We assessed the impact of TCT and lipopolysaccharide (LPS) on the functional integrity of the human airway mucosa by using in vitro airway mucosa models developed by co-culturing human tracheobronchial epithelial cells and human tracheobronchial fibroblasts on porcine small intestinal submucosa scaffold under airlift conditions. TCT and LPS either alone and in combination induced blebbing and necrosis of the ciliated epithelia. TCT and LPS induced loss of ciliated epithelial cells and hyper-mucus production which interfered with mucociliary clearance. In addition, the toxins had a disruptive effect on the tight junction organization, significantly reduced transepithelial electrical resistance and increased FITC-Dextran permeability after toxin incubation. In summary, the results indicate that TCT collaborates with LPS to induce the disruption of the human airway mucosa as reported for the hamster tracheal model.
Insights
Bordetella pertussis toxins, Tracheal Cytotoxin (TCT) and lipopolysaccharide (LPS), damage human airway mucosa. This study reveals how these toxins disrupt ciliated cells and impair mucociliary clearance, impacting respiratory health.
Area of Science:
- Microbiology and Immunology
- Respiratory Pathophysiology
Background:
- Bordetella pertussis causes whooping cough, characterized by respiratory mucosa disruption.
- Tracheal cytotoxin (TCT) is a key virulence factor recapitulating this pathology in animal models.
- Human data on TCT's effects are limited due to material scarcity.
Purpose of the Study:
- To assess the impact of TCT and lipopolysaccharide (LPS) on human airway mucosa integrity in vitro.
- To investigate the collaborative effect of TCT and LPS on respiratory epithelial cells.
Main Methods:
- Development of in vitro human airway mucosa models using co-cultured human tracheobronchial epithelial cells and fibroblasts on a porcine submucosa scaffold.
- Application of TCT and LPS, alone and in combination, to the models under airlift conditions.
- Assessment of epithelial integrity, cell viability, mucus production, tight junction organization, transepithelial electrical resistance (TEER), and permeability (FITC-Dextran).
Main Results:
- TCT and LPS induced ciliated epithelial blebbing, necrosis, and loss.
- Combined toxins led to hyper-mucus production, impairing mucociliary clearance.
- Toxins disrupted tight junctions, significantly reduced TEER, and increased epithelial permeability.
Conclusions:
- TCT collaborates with LPS to disrupt human airway mucosa, mirroring findings from hamster models.
- These toxins compromise the functional integrity of the human respiratory epithelium.
- The study provides crucial human in vitro data on B. pertussis virulence mechanisms.
