title: Inactivation of IL-6 and soluble IL-6 receptor by neutrophil derived serine proteases in cystic fibrosis.
abstract: The ability of IL-6 to signal via both membrane bound and soluble receptors is thought to explain the capacity of this cytokine to act in both the initiation and resolution of acute inflammatory responses. In cystic fibrosis (CF), poorly resolved neutrophillic inflammation of the lungs is a primary cause of morbidity and mortality. Expression of IL-6 has been reported to be low in CF lung secretions, despite ongoing inflammation, but the status of soluble IL-6 receptor (sIL-6R) in these patients is unknown. We hypothesised that sIL-6R may be an important potentiator of IL-6 activity in CF associated lung disease. IL-6, sIL-6R and sgp130 (a natural antagonist of responses mediated by the sIL-6R) were analysed by ELISA and Western blot in bronchoalveolar lavage fluid (BALF) from 28 paediatric CF patients and nine non-CF controls. Total cell counts in CF were four fold higher compared to controls (median: 1.4 x 10(6) cells/ml v. 0.35 x 10(6) cells/ml in controls) (p<0.001) and the infiltrate was dominated by neutrophils which were elevated by 89 fold (0.62 x 10(6) cells/ml v. 0.007 x 10(6) cells/ml in controls) (p<0.001). Other markers of inflammation such as IL-8 and MCP-1 were elevated 17.5 and 3.8 fold respectively (IL-8; median: 1122 pg/ml v. 64 pg/ml in controls, p<0.01 and MCP-1; median: 692 pg/ml v. 182 pg/ml in controls, p<0.05). IL-6, although present in 23/32 CF BALF specimens compared to 1/9 controls (p<0.01), was weakly expressed (median: 50 pg/ml). Expression of sIL-6R and sgp130 in CF was no different to control patients. We tested whether weak expression of all three molecules was due to degradation by CF BALF. Degradative activity was observed in association with BALF elastase activity and could be specifically blocked by serine protease inhibitors. Degradation of sIL-6R by purified serine proteases (elastase, cathepsin G and proteinase 3) was also observed leading to a loss of trans-signalling activity. Interestingly, sIL-6R was protected from proteolysis by interaction with IL-6. Our data identify and define a novel protease mediated deficiency of IL-6 signalling in the CF lung.
authors.affiliation: Department of Child Health, Cardiff University School of Medicine, Heath Park, Cardiff CF14 4XN, UK. mcgrealep@cf.ac.uk
authors.forename: Eamon P
authors.initials: EP
authors.lastname: McGreal
authors.forename: Philip L
authors.initials: PL
authors.lastname: Davies
authors.forename: Wendy
authors.initials: W
authors.lastname: Powell
authors.forename: Stefan
authors.initials: S
authors.lastname: Rose-John
authors.forename: O Bradley
authors.initials: OB
authors.lastname: Spiller
authors.forename: Iolo
authors.initials: I
authors.lastname: Doull
authors.forename: Simon A
authors.initials: SA
authors.lastname: Jones
authors.forename: Sailesh
authors.initials: S
authors.lastname: Kotecha
mesh_terms: D000328:Adult; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007223:Infant; D018836:Inflammation Mediators / Q000032:analysis / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism* / Q000494:pharmacology; D019272:Leukocyte Elastase / Q000378:metabolism; D009504:Neutrophils / Q000201:enzymology* / Q000473:pathology; D011485:Protein Binding / Q000502:physiology; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D019947:Receptors, Interleukin-6 / Q000737:chemistry / Q000378:metabolism*; D057057:Serine Proteases / Q000378:metabolism / Q000502:physiology*; D012995:Solubility
chemical_list: C508600:IL6 protein, human; C508603:IL6R protein, human; D018836:Inflammation Mediators; D015850:Interleukin-6; D019947:Receptors, Interleukin-6; D057057:Serine Proteases; D019272:Leukocyte Elastase