Non-typeable Haemophilus influenzae biofilm formation on air-liquid interface cultured primary human airway epithelia

Horton, Katie L., Allan, Raymond N., Coles, Janice L., Hueppe, Jana F., Cleary, David W., Jackson, Claire L. and Lucas, Jane S. (2021) Non-typeable Haemophilus influenzae biofilm formation on air-liquid interface cultured primary human airway epithelia. European Respiratory Journal, 58, [PA2354]. (doi:10.1183/13993003.congress-2021.PA2354).

Abstract

Introduction: defective motile cilia on primary ciliary dyskinesia (PCD) airway epithelia results in impaired mucociliary function and ultimately, bronchiectasis. Non-typeable Haemophilus influenzae (NTHi) is frequently isolated from PCD airways. It is unclear if biological pathways in response to NTHi infection differ in PCD, contributing to increased susceptibility to infection.

Aim: to develop a biologically representative model of mature NTHi infection of airway epithelia to investigate PCD airway epithelia-NTHi interactions.

Methods: air-liquid interface (ALI)-cultured primary nasal cells were used at 21-35 days post-ALI (n=30). Barrier properties were assessed by trans-epithelial electrical resistance (TEER), FITC-dextran passage and tight junctions staining. Cilia were assessed by high-speed video microscopy at 37˚C. NTHi PCD isolate was added to ALI cultures at multiplicity of infections (MOIs) 10, 50 or 100 for 1 hour and co-cultured for 3 days. TEER was measured pre- and post-infection. NTHi recoverability was assessed by colony forming units. Morphology was shown by scanning electron microscopy (SEM).

Results: 100% of ALI cultures succeeded. Tight barriers were formed. Ciliary beat pattern and frequency (mean 14Hz) were normal. Mean cilia coverage at day 29 was 38%. Following infection, TEER increased relative to pre-infection at all MOIs but maximum NTHi recoverability was at MOI 50. SEM showed widespread ciliation and NTHi biofilm formation at MOI 50.

Conclusions: a successful model of mature NTHi infection of human primary airway was established. The model will be used with PCD, and other airway disease, epithelia to investigate host-pathogen interactions.

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Contributors

Author: David W. Cleary

Author: Claire L. Jackson

Author: Jane S. Lucas

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