Bronchial epithelial cell lines and primary nasal epithelial cells from cystic fibrosis respond differently to cigarette smoke exposure  Mark Thomas Shaw Williams, Francine de Courcey, David Comer, Joseph S. Elborn, Madeleine Ennis  Journal of Cystic Fibrosis  Volume 15, Issue 4, Pages 467-472 (July 2016) DOI: 10.1016/j.jcf.2015.11.002 Copyright © 2015 Terms and Conditions

Fig. 1 Viability of non-CF and CF AECs following exposure to CSE. 16HBE14o− (dashed line), and CFBE41o− (solid line) cell lines (a), and NECs derived from healthy controls (open squares), R117H/F508del heterozygous CF patients (open circles) and F508del homozygous CF patients (open triangles) (b), were incubated with a range of CSE concentrations (1.5%–50%) for 4h. The MTT proliferation assay was carried out on these cells. Viability of the cells was then expressed as a percentage of the absorbance displayed by the controls (relative viability). Data are median±IQR and represent N=3 independent experiments for the non-CF and CF AECs each. Journal of Cystic Fibrosis 2016 15, 467-472DOI: (10.1016/j.jcf.2015.11.002) Copyright © 2015 Terms and Conditions

Fig. 2 CSE effects on inflammatory responses displayed by AECs at baseline and following inflammatory stimuli are cell model specific. 16HBE14o− (a), CFBE41o− (b) cell lines, and NECs derived from healthy controls (c), R117H/F508del heterozygous CF patients (d) and F508del homozygous CF patients (e), were exposed/unexposed to 5% CSE for 4h. Cells were then stimulated with various pro-inflammatory stimuli (as mentioned previously) for 24h. Basal and induced IL-8 release was then determined by ELISA of harvested supernatants. Data are mean±SEM and represent N=6 independent experiments for each cell line, and N=6, N=3–7 and N=5 independent experiments for the healthy, R117H/F508del heterozygous and F508del homozygous NECs respectively. Paired sample t test, *p<0.05, and **p<0.01. Journal of Cystic Fibrosis 2016 15, 467-472DOI: (10.1016/j.jcf.2015.11.002) Copyright © 2015 Terms and Conditions

Fig. 3 The EGFR pathway plays a role in basal and CSE induced IL-8 release from non-CF and CF AECs. 16HBE14o− (a), CFBE41o− (b) cells, and NECs derived from healthy individuals (c) and F508del homozygous CF patients (d) were incubated with increasing concentrations of the specific EGFR kinase inhibitor AG-1478 (50nM–1μM) in the presence and absence of 5% CSE for 4h. Constitutive and CSE induced IL-8 release was then determined by ELISA of harvested supernatants. IL-8 release from these cells was then expressed as a percentage of IL-8 secretion exhibited by the media control, IL-8 (% control). Data are median and represent N=4 independent experiments for each cell line and N=3 independent experiments for both the healthy and CF NECs. Paired Student t-test, ***p<0.001. Journal of Cystic Fibrosis 2016 15, 467-472DOI: (10.1016/j.jcf.2015.11.002) Copyright © 2015 Terms and Conditions