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An Inverse Transwell Assay for Airway Mucus Barrier Function Reveals both Virus- and Mucin-Specific Impacts on
Maria Corkran1,2, Elizabeth M Engle2,3, Heng Pan4,5
1Department of Cell Biology and Molecular Genetics, Maryland Pathogen Research Institute, University of Maryland, College Park, MD, USA.
Biorxiv : the Preprint Server for Biology
|July 29, 2026
Summary
Airway mucus traps respiratory viruses, but its effectiveness varies by virus type. MUC5AC mucin is crucial for mucus structure and antiviral defense, impacting how effectively viruses like influenza penetrate the mucus barrier.
Area of Science:
- * Respiratory immunology
- * Virology
- * Mucosal immunology
Background:
- * Respiratory viruses cause significant global morbidity and mortality.
- * Airway mucus, rich in mucins, acts as a primary defense against respiratory viruses.
- * Altered mucin expression in chronic lung disease may affect susceptibility to viral infections.
Purpose of the Study:
- * To evaluate the barrier function of human airway mucus against different respiratory viruses.
- * To investigate the role of specific mucins (MUC5B and MUC5AC) in mucus antiviral properties.
- * To establish an in vitro model for studying mucus penetration by viruses.
Main Methods:
- * Developed an inverse Transwell assay (ITA) to assess mucus barrier function.
- * Tested mucus from human airway epithelial (HAE) cultures against influenza (IAV), rhinovirus (RV), adenovirus, and parainfluenza virus.
- * Utilized CRISPR/Cas9 to create HAE cultures deficient in MUC5B or MUC5AC for infection studies and mucus analysis.
Main Results:
- * Mucus exhibited differential inhibition of viral penetration, with influenza virus (IAV) being most inhibited.
- * Mucin-deficient HAE cultures showed higher IAV titers, indicating MUC5B and MUC5AC contribute to antiviral defense.
- * MUC5AC-depleted mucus allowed more efficient IAV penetration and displayed larger pore structures, suggesting a role in mucus barrier integrity.
Conclusions:
- * Mucus-mediated viral restriction is virus-specific.
- * MUC5AC plays a key role in maintaining mucus structure and its ability to impede viral infection.
- * The ITA is a valuable tool for studying virus-mucus interactions and the impact of altered mucus composition.
