Fig 1.
Influence of IL-27 on the inflammatory response in sterile zymosan-induced peritonitis.
IL-27 was either given together with zymosan (zymosan +IL-27) or 12 h before zymosan application into the peritoneum (zymosan +preIL-27). The control group received zymosan only. A. Representative FACS plots for peritoneal neutrophils and macrophages isolated from the peritoneal lavage 12h after induction of peritonitis. GR1+CD11b+ cells are defined as neutrophils, CD11b+ single positive cells as macrophages. Cell frequencies within the neutrophil gate are depicted. B. Absolute number of neutrophils in the peritoneal lavage at various time points after zymosan injection. Results are pooled from four independent experiments and represent mean ± S.E.M (n = 4–12). C. Absolute number of macrophages in the peritoneal lavage at various time points after zymosan injection. Pooled data from four independent experiments, mean ± S.E.M. (n = 4–12). Mann–Whitney test was used to compare between groups, *p<0.05, ** p<0.01.
Fig 2.
IL-27 pre-treatment does not affect the cell numbers of T, B, NK and dendritic cells in the peritoneal lavage.
Cells in the peritoneal lavage were collected 12 h after the administration of zymosan with or without pre-treatment with rIL-27 and measured by FACS analysis. Results from three different experiments are combined, given are means ± S.D (n = 4–9). Mann–Whitney test was used to compare between groups, *p<0.05, ** p<0.01, ***p<0.005.
Fig 3.
IL-27 pre-treatment decreases neutrophil accumulation in the murine blood and temporarily in the bone marrow during zymosan-induced peritonitis.
Dot plots show the gating for neutrophils in the FACS analyses of blood cells and bone marrow cells (at at 12h and 4h after zymosan injection, respectively). Cell frequencies within the gate are given for the exemplary FACS plots. A. Absolute numbers of blood neutrophils in the blood of vehicle-treated animals (vehicle) 12h after injection and animals with or without pre-treatment with rIL-27 monitored 4–48h after injection of zymosan. Results from four different experiments are combined; given are means ± S.E.M (n = 8–11). B. Absolute numbers of neutrophils in the bone marrow of vehicle-treated animals (12h after vehicle injection) and 4–48h after injection of zymosan with or without pre-treatment with rIL-27. Results from four different experiments are combined; given are means ± S.E.M (n = 8–11). Mann–Whitney test was used to compare between groups, *p<0.05, ** p<0.01, ***p<0.005.
Fig 4.
Comparable levels of cytokines in the peritoneal exudate.
12 h after induction of peritonitis the peritoneal cavity was flushed with 5 ml PBS and levels of the indicated cytokines were measured by multiplex bead array assay. Results from three different experiments are combined; given are means ± S.D (n = 6–10). Mann–Whitney test was used to compare between groups.
Fig 5.
Decreased levels of KC, MCP-1 and MIP-1α in the peritoneal lavage after IL-27 pretreatment.
12 h after induction of peritonitis or vehicle injection, the peritoneal cavity was flushed with 5ml PBS and levels of the indicated chemokines were measured by multiplex bead array assay. Results are pooled from three independent experiments; given are means ± S.D (n = 6–8). Mann–Whitney test was used to compare between groups, *p<0.05, ** p<0.01, ***p<0.005.