Flow cytometry.
Peritoneal lavage cells were surface stained with the antibodies listed in Supplementary Table 1. Flow cytometry data were acquired in a FACS Canto II Cytometer (BD), at the Instituto de Higiene, Universidad de la República, and then analysed using the FlowJo software package. Single cells were selected on the basis of FSC-H and FSC-A, and then viable cells selected using the Live/Dead GreenTM viability probe. Within live cell singlets, eosinophils and neutrophils were defined as SiglecFhi and Ly6G+ cells respectively. B cells (CD19+) were excluded from SiglecF- Ly6G- cells. SiglecF- Ly6G-CD19- cells were divided into monocytes (Ly6Chi) and non-monocytes (Ly6C-). The second category was further divided into small peritoneal macrophages and dendritic cells (SPM/DC; MHCII+F4/80-), MHCII-F4/80med macrophages, and large peritoneal macrophages (LPM; MHCII- F4/80high) (26, 27). The gating strategy is summarised in supplementary Figure S1.
Statistical analysis .
Data were analysed by the non-parametric method of Kruskal-Wallis (28), followed by the post-hoc multiple comparison test described by Conover (29) and the Benjamini-Hochberg correction for controlling the false discovery rate (30). The symbols *, ** and *** represent p-values less than 0.05, 0.01 and 0.001 respectively.