title: Corticosteroid effects on COPD alveolar macrophages: dependency on cell culture methodology.
abstract: It is unclear whether cell culture methodology affects the corticosteroid sensitivity of chronic obstructive pulmonary disease (COPD) alveolar macrophages. We compared the effect of a short and a long isolation procedure on corticosteroid inhibition of lipopolysaccharide (LPS) stimulated cytokine release from COPD alveolar macrophages. We also investigated signalling pathways associated with macrophage activation during cell isolation. Macrophages cultured using a short isolation protocol released higher unstimulated levels of tumour necrosis factor (TNF)-α and chemokine C-X-C motif ligand (CXCL) 8; these macrophages were less sensitive to corticosteroid inhibition of LPS stimulated TNF-α and CXCL8 release when compared to a long isolation procedure. This was associated with increased p38 mitogen activated kinase (MAPK) activation. The p38 MAPK inhibitor, BIRB-796, significantly reduced unstimulated cytokine release. A key finding of this study was that both cell culture methods showed no difference in the corticosteroid sensitivity between COPD and control macrophages. We conclude that the culture of alveolar macrophages using a short isolation procedure alters cytokine production through p38 MAPK activation; this is associated with a change in corticosteroid sensitivity.
authors: The University of Manchester, Manchester Academic Health and Science Centre, University Hospital of South Manchester Foundation Trust, Respiratory and Allergy Centre, Institute of Inflammation and Repair, Faculty of Medical and Human Sciences, Manchester M23 9LT, UK. Electronic address: Andrew.Higham@manchester.ac.uk.
authors: Andrew
authors: A
authors: Higham
authors: The University of Manchester, Manchester Academic Health and Science Centre, University Hospital of South Manchester Foundation Trust, Respiratory and Allergy Centre, Institute of Inflammation and Repair, Faculty of Medical and Human Sciences, Manchester M23 9LT, UK.
authors: Simon
authors: S
authors: Lea
authors: The University of Manchester, Manchester Academic Health and Science Centre, Central Manchester University Hospitals Foundation Trust, Centre for Endocrinology and Diabetes, Institute for Human Development, Faculty of Medical and Human Sciences, Manchester M13 9PT, UK.
authors: David
authors: D
authors: Ray
authors: The University of Manchester, Manchester Academic Health and Science Centre, University Hospital of South Manchester Foundation Trust, Respiratory and Allergy Centre, Institute of Inflammation and Repair, Faculty of Medical and Human Sciences, Manchester M23 9LT, UK.
authors: Dave
authors: D
authors: Singh
mesh_terms: D000368:Aged; D015153:Blotting, Western; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000378:metabolism; D016207:Cytokines / Q000276:immunology* / Q000378:metabolism; D003907:Dexamethasone / Q000276:immunology* / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005938:Glucocorticoids / Q000276:immunology* / Q000494:pharmacology; D006801:Humans; D016209:Interleukin-8 / Q000276:immunology / Q000378:metabolism; D008070:Lipopolysaccharides / Q000276:immunology / Q000494:pharmacology; D016676:Macrophages, Alveolar / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009281:Naphthalenes / Q000276:immunology / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects / Q000276:immunology; D029424:Pulmonary Disease, Chronic Obstructive / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D011720:Pyrazoles / Q000276:immunology / Q000494:pharmacology; D051996:Transcription Factor RelA / Q000276:immunology / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism; D010671:Phenylurea Compounds
chemical_list: D017077:Culture Media, Conditioned; D016207:Cytokines; D003907:Dexamethasone; D005938:Glucocorticoids; D016209:Interleukin-8; D008070:Lipopolysaccharides; D009281:Naphthalenes; D011720:Pyrazoles; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D048051:p38 Mitogen-Activated Protein Kinases; C452139:doramapimod; D010671:Phenylurea Compounds
keywords: Alveolar macrophage; Chronic obstructive pulmonary disease; Corticosteroids; Cytokines; Inflammation; p38 MAPK