Abstract
IntroductionDifferentiated paediatric epithelial cells can be used to study the role of epithelial cells in asthma. Nasal epithelial cells are easier to obtain and may act as a surrogate for bronchial epithelium in asthma studies. We assessed the suitability of nasal epithelium from asthmatic children to be a surrogate for bronchial epithelium using air-liquid interface cultures.MethodsPaired nasal and bronchial epithelial cells from asthmatic children (n = 9) were differentiated for 28 days under unstimulated and IL-13-stimulated conditions. Morphological and physiological markers were analysed using immunocytochemistry, transepithelial-electrical-resistance, Quantitative Real-time-PCR, ELISA and multiplex cytokine/chemokine analysis.ResultsPhysiologically, nasal epithelial cells from asthmatic children exhibit similar cytokine responses to stimulation with IL-13 compared with paired bronchial epithelial cells. Morphologically however, nasal epithelial cells differed significantly from bronchial epithelial cells from asthmatic patients under unstimulated and IL-13-stimulated conditions. Nasal epithelial cells exhibited lower proliferation/differentiation rates and lower percentages of goblet and ciliated cells when unstimulated, while exhibiting a diminished and varied response to IL-13.ConclusionsWe conclude that morphologically, nasal epithelial cells would not be a suitable surrogate due to a significantly lower rate of proliferation and differentiation of goblet and ciliated cells. Physiologically, nasal epithelial cells respond similarly to exogenous stimulation with IL-13 in cytokine production and could be used as a physiological surrogate in the event that bronchial epithelial cells are not available.
Highlights
Differentiated paediatric epithelial cells can be used to study the role of epithelial cells in asthma
Following IL-13 stimulation, there was a significant difference in Transepithelial Electrical Resistance measurements (TEER) values on days 7 (p = 0.02) & 14 (p = 0.02) between PBECs and profiles of nasal differentiated ALI cultures (PNECs) with the difference in resistance becoming similar by days 21 & 28 [Figure 1B]
There was no significant difference in the total cell number under IL-13 stimulated conditions [PBECs [mean 5.36105 cells/ml (SD 0.9); PNECs: mean 4.26105 cells/ml (SD 1.1)] [Table 2]
Summary
Differentiated paediatric epithelial cells can be used to study the role of epithelial cells in asthma. Nasal epithelial cells are easier to obtain and may act as a surrogate for bronchial epithelium in asthma studies. Bronchial epithelial airway remodelling is characterised by goblet cell hyperplasia, reduced ciliated cell numbers and mucus hypersecretion [5], defective repair and proliferation [6], increased basal cell number [7] and impaired barrier function [8,9]. Differentiated ALI cultures using the Transwell system have recently been shown to be an authentic model representing the airway epithelium ex vivo [10]. Differentiated ALI cultures from healthy subjects displayed a polarised pseudostratified multi-layered epithelium comprising basal, ciliated and goblet cells whereas cultures from asthmatic subjects displayed a dysfunctional epithelium consistent with the asthmatic airway in vivo [5]. The cultures closely mimic in vivo airway epithelial physiology in terms of cilia coverage and cilial beating, mucus production and formation of intact tight junctions [5,11]
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