Fig. 1

Ibrutinib inhibits LTA and S. pneumoniae-induced macrophage and PMN activation in vitro. a RAW264.7 macrophages were stimulated for 5 min with 10 μg/mL lipoteichoic acid (LTA) in the presence of 2500 nM Ibrutinib in IMDM containing 0,1% DMSO or vehicle (0.1% DMSO in IMDM). Cell lysates were analyzed by Western blot for total Bruton’s tyrosine kinase (Btk), Tyrosine (Y) 223 phosphorylated Btk and β-actin. b RAW 264.7 cells were stimulated with 0.01, 0.1 and 1 μg/mL LTA or (c) UV-killed S. pneumoniae strain 6303 (cell:bacterium ratio 1:100) (n = 8 wells group) for 24 h in the presence of 2500 nM Ibrutinib or vehicle. The concentration of tumor necrosis factor (TNF) in supernatant is depicted. d Bone marrow polymorphonuclear cells (PMN) were stimulated with LTA (10 μg/ml) or UV-killed S. pneumoniae strain 6303 (cell:bacterium ratio 1:100) (n = 8 wells group) for 1 h in the presence of 2500 nM Ibrutinib or vehicle. CD11b expression was determined by flow cytometry. Data are represented as the mean of each group with error bars representing SD. *p < 0.05, ***P < 0.001 (Mann–Whitney U-test). Data represent results from one (d), two (a) or three (b and c) independent experiments