Fig 5. Different IL-1β production in macrophages and neutrophils from Ncf1-/- and wild type mice and the activity of neutrophil elastase to process pro-IL-1β.
Thioglycollate-elicited macrophages (1x105) and neutrophils (1x106) from Ncf1-/- and WT mice were infected with M. marinum at MOI:1, the IL-1β release was then determined by ELISA (A and B). Differentiated THP-1 cells were left uninfected (control) or pretreated by PBS and DPI then infected for 4 h with M. marinum at MOI:1, 18 h later, pro-IL-1β (p34) and mature-IL-1β (p17) (C) were analyzed by immunoblot. NE activity of human granulocytes (1x106) were left untreated or pre-treated by NE inhibitor and then stimulated with PMA (100nM) as a positive control and M. marinum (MOI: 5) (D). Recombinant pro-IL1β (50 ng) was treated by NE (2x10-4 U) with and without NE inhibitor, and mature-IL-1β (p17) was analyzed by immunoblot (E). Data represent mean ± sd. The experiments were analyzed with Kruskal-Wallis analysis and repeated 2 times with similar results. *p < 0.05; **p < 0.005.