Fig. 9

CLEC-2-podoplanin axis regulates the inflammatory reaction in vitro and in vivo. a–c Bone marrow derived macrophages (BMDM) were incubated in the presence of LPS (1 μg/ml) or media for 18 h. a Podoplanin expression assessed by flow cytometry gated on annexin V-negative BMDM. b,c LPS-treated BMDM were incubated with anti-podoplanin antibody (mAb 8.1.1) or Syrian hamster IgG (IgG control) for 24 h. b TNF-α secretion and c iNOS expression was assessed. d–o WT mice were subjected to cecal ligation and puncture (CLP) for 24 h. Two hours post cecal ligation and puncture (CLP), mAb 8.1.1 (100 μg/mouse) or Syrian hamster IgG (IgG control) were injected i.v. and d clinical score assessed at 24h. e CXCL-1, f CCL2, g IL-6, h TNF-α and i IL-10 were measured in peritoneal lavage 24 h after CLP. Immune cell recruitment to the peritoneal cavity was assessed by flow cytometry. Total number of j CD45+ cells k CD45 + CD11b+ myeloid cells, l neutrophils (CD45+CD11b+Ly6G+) and m macrophages (CD45 + CD11b + F4/80+) in PLF was measured. n Bacteria in the PLF was assessed 24h post CLP. Colony unit formation (CFU) were counted and adjusted to the PLF volume. o Hemoglobin levels measured in the peritoneal lavage fluid 24 h post CLP. Data are means ± s.d. (n = 7). Differences analyzed by Mann–Whitney U-test. NS non-significant, *p < 0.05, **p < 0.01, ***p < 0.001