Flow cytometric analysis of peritoneal leukocytes populations following BZ5B8 administration in vivo.

Female C57BL/6 mice were injected with anti-CCRL2 antibody BZ5B8 or isotype control intraperitoneally (IP), followed by human CXCL8 or PBS IP injection to induce the model. Peritoneal lavages were harvested 4 h later for flow cytometry analysis. (A) Percentages of neutrophils (left) out of total CD45+ leukocytes in peritoneal lavages and surface expression of CXCR2 on neutrophils (right) are reported for each treatment group. (B) Percentages of dendritic cells (left) and monocytes (right) out of total CD45+ leukocytes in peritoneal lavages are reported for each treatment group. (C) Mouse anti-CCRL2 antibody BZ5B8 plasma concentration (Mean ± SD) 24 h after IP dose. (D) CCRL2 expression is shown on neutrophils (left), dendritic cells (middle) and monocytes (right) in control mice stimulated with CXCL8 or PBS control. (E) Representative histogram of CCRL2 expression on dendritic cells (grey), non-myeloid cells (CD11b-, Ly6G-) (orange), monocytes/macrophages (blue) or neutrophils (red) compared to FMO control in peritoneal lavages from CXCL8 stimulated mice. (A-B, D) Statistical analyses were performed using the Brown-Forsythe and Welch ANOVA with Dunnett’s T3 multiple comparisons test, and a unpaired t test with Welch’s correction. Statistical analyses used an alpha threshold of 0.05. Results are plotted as the Mean ± SEM. **P<0.01.