title: Bronchial epithelial cell lines and primary nasal epithelial cells from cystic fibrosis respond differently to cigarette smoke exposure.
abstract: The effects of cigarette smoke extract (CSE) on airway epithelial cells (AECs) from cystic fibrosis (CF) and non-cystic fibrosis (non-CF) individuals are not fully understood. It has been suggested that CSE modulates inflammatory cytokine release from AECs by modulating the epidermal growth factor receptor (EGFR) pathway; these pathways could reveal novel therapeutic targets. We compared the effect of CSE pre-incubation on IL-8 release from CF and non-CF bronchial epithelial cell lines, and separately, with primary nasal epithelial cells (NECs) retrieved from CF and non-CF individuals. We also determined if the EGFR pathway regulates IL-8 release by LPS or cytomix in non-CF and CF AECs at baseline and following CSE exposure. CF and non-CF cell lines, NECs derived from both CF patients (R117H heterozygous and F508del homozygous), and from healthy subjects, were cultured in the presence or absence of CSE, and subsequently exposed to inflammatory stimuli. In cell lines CSE significantly reduced IL-8 release following inflammatory challenge. Conversely, CSE pre-treatment was pro-inflammatory in primary NECs. In NECs from control subjects, CSE increased cytomix and LPS induced IL-8 release, and for the R117H heterozygous NEC cultures, CSE enhanced basal IL-8 release. Cytomix and LPS induced IL-8 release from F508del homozygous NEC cultures was further heightened following CSE pre-treatment. EGFR inhibition mitigated IL-8 release from immortalised and primary non-CF and CF AECs, suggesting that constitutive and CSE elicited IL-8 release from AECs is partly regulated via the EGFR pathway. This study demonstrates the importance of the EGFR cascade in the regulation of constitutive and CSE induced inflammatory mediator release from immortalised and primary AECs. Moreover, it clearly highlights the significance of using primary cells to confirm results obtained from immortalised cell studies, as these model systems may respond very differently to the stimuli under investigation.
authors.affiliation: Centre for Infection and Immunity, Queen's University Belfast, Belfast BT9 7AE, Northern Ireland, United Kingdom; Institute for Infection, Immunity and Inflammation, University of Glasgow, Glasgow, G12 8TA Scotland, United Kingdom. Electronic address: mark.williams@glasgow.ac.uk.
authors.forename: Mark Thomas Shaw
authors.initials: MT
authors.lastname: Williams
authors.affiliation: Centre for Infection and Immunity, Queen's University Belfast, Belfast BT9 7AE, Northern Ireland, United Kingdom.
authors.forename: Francine
authors.initials: F
authors.lastname: de Courcey
authors.affiliation: Centre for Infection and Immunity, Queen's University Belfast, Belfast BT9 7AE, Northern Ireland, United Kingdom.
authors.forename: David
authors.initials: D
authors.lastname: Comer
authors.affiliation: Centre for Infection and Immunity, Queen's University Belfast, Belfast BT9 7AE, Northern Ireland, United Kingdom.
authors.forename: Joseph S
authors.initials: JS
authors.lastname: Elborn
authors.affiliation: Centre for Infection and Immunity, Queen's University Belfast, Belfast BT9 7AE, Northern Ireland, United Kingdom.
authors.forename: Madeleine
authors.initials: M
authors.lastname: Ennis
mesh_terms: D001980:Bronchi / Q000473:pathology; D002460:Cell Line; D000073865:Cigarette Smoking / Q000276:immunology; D003550:Cystic Fibrosis / Q000276:immunology*; D004847:Epithelial Cells / Q000276:immunology*; D066246:ErbB Receptors / Q000378:metabolism*; D006801:Humans; D007249:Inflammation / Q000139:chemically induced / Q000276:immunology; D016209:Interleukin-8 / Q000378:metabolism*; D009297:Nasal Mucosa / Q000473:pathology; D015398:Signal Transduction; D012906:Smoke / Q000009:adverse effects*
chemical_list: D016209:Interleukin-8; D012906:Smoke; D066246:ErbB Receptors
keywords: Airway; Bronchial; CSE; Cells; Cytokine; EGFR; Epithelial; F508del; Healthy; IL-8; Immortalised; Inflammation; Nasal; Patients; Primary; R117H; Release