title: Smoking enhances the proinflammatory effects of nucleotides on cytokine release from human lung.
abstract: Nucleotides have effects on immune cells which are complex but generally proinflammatory, and have been suggested to play a role in smoking-related lung diseases. However, there have been no studies directly measuring functional responses to nucleotides in human lungs taken from smokers. We used fragments of post mortem human lung from smokers and non-smokers, incubated them with a range of nucleotides (4-1000 µM) in the presence of lipopolysaccharide (LPS; 10 µg/ml) for 24 hours and measured cytokines (IL-1β, IFNγ, IL-17, TNFα, IL-6, IL-8, IL-2 and IL-10) in the supernatants using multiplex immunoassays. Although the basal cytokine levels in the smokers were generally higher in the smokers than the non-smokers, there were no significant differences in either the basal release or the LPS-stimulated release of any of the cytokines when lungs from smokers and non-smokers were compared. There were no significant effects of ATP, ADP, AMP, UTP, α,β-methylene-ATP, P1, P4-diATP, 2-methylthio-ATP or Bz-ATP on the release of cytokines from the lungs. However, the stable ATP analogue ATPγS increased the release of IL-1β and IFNγ, and the effect was greatly increased in lungs from smokers. In non-smokers but not in smokers ATPγS increased the release of IL-17. Overall these results clearly demonstrate for the first time that in normal human lung a stable ATP analogue can enhance LPS-induced pro-inflammatory cytokine release, and that these effects are greatly altered by a prior history of smoking. This provides strong support for the suggestion that nucleotides are involved in the pathogenesis of smoking-related diseases.
authors: Faculty of Health and Medical Sciences, University of Surrey, Guildford, Surrey, United Kingdom.
authors: Kylie
authors: K
authors: Belchamber
authors: Fibrosis Discovery Performance Unit, GlaxoSmithKline, Stevenage, Hertfordshire, United Kingdom.
authors: David A
authors: DA
authors: Hall
authors: Faculty of Health and Medical Sciences, University of Surrey, Guildford, Surrey, United Kingdom.
authors: Susanna M O
authors: SM
authors: Hourani
mesh_terms: D016022:Case-Control Studies; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008070:Lipopolysaccharides / Q000633:toxicity; D008168:Lung / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D009711:Nucleotides / Q000494:pharmacology*; D012907:Smoking / Q000009:adverse effects*
chemical_list: D016207:Cytokines; D008070:Lipopolysaccharides; D009711:Nucleotides