Cigarette smoke modifies neutrophil chemotaxis, neutrophil extracellular trap formation and inflammatory response-related gene expression

• Main Authors: Chapple, I.L.C (Author), Cooper, P.R (Author), Hirschfeld, J. (Author), Matthews, J.B (Author), Milward, M.R (Author), White, P.C (Author), Wright, H.J (Author)
• Format: Article
• Language: English
• Published: Blackwell Munksgaard 2018
• Subjects: apoptosis; Apoptosis; chemotaxis; Chemotaxis; cigarette smoke; cotinine; Cotinine; drug effect; extracellular trap; Extracellular Traps; flow cytometry; Flow Cytometry; gene expression; Gene Expression; human; Humans; inflammation; metabolism; neutrophil; neutrophils; Neutrophils; nicotine; Nicotine; periodontitis; reactive oxygen metabolite; Reactive Oxygen Species; real time polymerase chain reaction; Real-Time Polymerase Chain Reaction; smoke; Smoke; smoking; Smoking; thiocyanate; Thiocyanates; thiocyanic acid derivative
• Online Access: View Fulltext in Publisher

 Background and Objective: Cigarette smoking is a major risk factor for periodontitis, and smoking perturbs neutrophil reactive oxygen species production. This study tested the hypothesis that cigarette smoke extract (CSE) and its components/metabolites nicotine, cotinine and thiocyanate (SCN-), may influence neutrophil functions. Material and Methods: Chemotaxis was assessed in neutrophils pre-treated with CSE using real-time video microscopy. Neutrophil extracellular trap (NET) release in response to CSE, nicotine, cotinine, SCN- as well as to phorbol 12-myristate-13-acetate and hypochlorous acid following pre-treatment with CSE, nicotine, cotinine or SCN- was assessed using fluorescence-based assays. The impact of CSE and SCN- treatment on neutrophil respiratory burst- and inflammation-related gene expression (NFKBIE, DNAJB1, CXCL8, NCF1, NCF2, CYBB) was determined by real-time polymerase chain reaction. Results: Both CSE and SCN- pre-treatment inhibited phorbol 12-myristate-13-acetate-stimulated NET release. Additionally, SCN- inhibited hypochlorous acid-stimulated NET formation, while SCN- alone stimulated NET release. Overall, neutrophils pre-treated with CSE exhibited reduced speed, velocity and directionality relative to untreated neutrophils. Although CSE and SCN- promoted DNAJB1 expression, increased redox-related gene expression was only detected in response to SCN-. Conclusion: These results suggest that CSE can alter ex vivo neutrophil activation by mechanisms independent of SCN- and nicotine, and SCN- may contribute to the perturbed innate immune responses observed in smokers. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd