scholarly journals Characterisation of morphological differences in well-differentiated nasal epithelial cell cultures from preterm and term infants at birth and one-year

2018 ◽  
Author(s):  
Helen E. Groves ◽  
Hong Guo-Parke ◽  
Lindsay Broadbent ◽  
Michael D. Shields ◽  
Ultan F. Power

AbstractBackgroundInnate immune responses of airway epithelium are important defences against respiratory pathogens and allergens. Newborn infants are at greater risk of severe respiratory infections compared to older infants. However, very little is known regarding human neonatal airway epithelium immune responses and whether age-related morphological and/or innate immune changes contribute to the development of airway disease.MethodsWe collected nasal epithelial cells from 41 newborn infants (23 term, 18 preterm) within 5 days of birth. Repeat sampling was achieved for 24 infants (13 term, 11 preterm) at a median age of 12.5 months. Morphologically and physiologically authentic well-differentiated primary paediatric nasal epithelial cell (WD-PNEC) cultures were generated and characterised using light microscopy and immunofluorescence.ResultsWD-PNEC cultures were established for 15/23 (65%) term and 13/18 (72%) preterm samples at birth, and 9/13 (69%) term and 8/11 (73%) preterm samples at one-year. Newborn and infant WD-PNEC cultures demonstrated extensive cilia coverage, mucous production and tight junction integrity. Newborn WD-PNECs took significantly longer to reach full differentiation and were noted to have much greater proportions of goblet cells compared to one-year repeat WD-PNECs. No differences were evident in ciliated/goblet cell proportions between term- and preterm-derived WD-PNECs at birth or one-year old.ConclusionWD-PNEC culture generation from newborn infants is feasible and represents a powerful and exciting opportunity to study differential innate immune responses in human airway epithelium very early in life.

2008 ◽  
Vol 294 (6) ◽  
pp. L1068-L1075 ◽  
Author(s):  
Jonathan L. Koff ◽  
Matt X. G. Shao ◽  
Iris F. Ueki ◽  
Jay A. Nadel

Toll-like receptors (TLRs) are critical for the recognition of inhaled pathogens that deposit on the airway epithelial surface. The epithelial response to pathogens includes signaling cascades that activate the EGF receptor (EGFR). We hypothesized that TLRs communicate with EGFR via epithelial signaling to produce certain innate immune responses. Airway epithelium expresses the highest levels of TLR2, TLR3, TLR5, and TLR6, and here we found that ligands for these TLRs increased IL-8 and VEGF production in normal human bronchial epithelial cells. These effects were prevented by treatment with a selective inhibitor of EGFR phosphorylation (AG-1478), a metalloprotease (MP) inhibitor, a reactive oxygen species (ROS) scavenger, and an NADPH oxidase inhibitor. In an airway epithelial cell line (NCI-H292), TNF-α-converting enzyme (TACE) small interfering RNA (siRNA) was used to confirm that TACE is the MP involved in TLR ligand-induced IL-8 and VEGF production. We show that transforming growth factor (TGF)-α is the EGFR ligand in this signaling cascade by using TGF-α neutralizing antibody and by showing that epithelial production of TGF-α occurs in response to TLR ligands. Dual oxidase 1 (Duox1) siRNA was used to confirm that Duox1 is the NADPH oxidase involved in TLR ligand-induced IL-8 and VEGF production. We conclude that multiple TLR ligands induce airway epithelial cell production of IL-8 and VEGF via a Duox1→ ROS→ TACE→ TGF-α→ EGFR phosphorylation pathway. These results show for the first time that multiple TLRs in airway epithelial cells produce innate immune responses by activating EGFR via an epithelial cell signaling cascade.


PLoS ONE ◽  
2012 ◽  
Vol 7 (3) ◽  
pp. e32728 ◽  
Author(s):  
William A. Rose ◽  
Chris L. McGowin ◽  
Rae Ann Spagnuolo ◽  
Tonyia D. Eaves-Pyles ◽  
Vsevolod L. Popov ◽  
...  

2019 ◽  
Vol 1 (1A) ◽  
Author(s):  
Patricia J. Coulter ◽  
Helen Groves ◽  
Lindsay Broadbent ◽  
Jonathan Dean Coey ◽  
Michael D. Shields ◽  
...  

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