Fig 2: Brilliant Blue G (BBG) treatment does not affect the distribution of splenic leukocytes from SOD1G93A mice.Splenic leukocytes from BBG- or saline-treated SOD1G93A mice at end-stage were labelled with fluorochrome-conjugated mAbs. The percentage of (A) B cells (CD19+CD3−), (B) T cells (CD3+CD19−), (C) CD8+ T cells (CD8+CD3+CD19−), (D) CD4+ T cells (CD4+CD3+), (E) regulatory T (Treg) cells (CD25+CD4+CD3+), (F) dendritic cells (DCs; CD11b±CD11c+), (G) macrophages (MOs; CD11b+CD11c−Ly6G−) and (H) polymorphonuclear neutrophils (PMNs; CD11b+CD11c−Ly6G+) of the total CD45+ leukocyte population were determined by flow cytometry. Results shown as means ± SD, n = 6 (male and female). Symbols represent individual animals.

Fig 3: Characterization of PILR-α-/- and PILR-β1-/-neutrophils for surface expression of molecules involved in extravasation.Ly6G+ bone marrow neutrophils from WT, PILR-α-/- or PILRβ1-/- mice were analyzed for indicated antigens by FACS. Representative histograms of at least three independent experiments are shown in (A) for PILR-α-/- and (C) for PILR-β1-/- mice, with expression levels quantified in (B) and (D), respectively. Solid tracks, specific staining; dotted tracks, isotype control. Groups were compared by 2-tailed t-test. (E) qRT-PCR analysis of RNA extracted from peripheral blood of WT (n = 9) or PILR-α-/- (n = 5) or PILR-β1-/- (n = 7) mice for the indicated genes. Groups were analyzed by 1-way ANOVA followed by Holm-Sidak method for multiple comparisons. Error bars, SEM. ****p<0.001.10.7554/eLife.47642.010Figure 1—source data 1.Source data for Figure 1B and D.10.7554/eLife.47642.011Figure 1—source data 2.Source data for Figure 1E.

Fig 4: Reduction number of MDSCs alleviates inflammation in asthmatic mice. Asthma was induced in mice that were then treated with gemcitabine as described in the Materials and Methods section. The single-cell suspensions of the lungs in mice were tested in terms of CD11b+Ly6G-Ly6Chigh M-MDSC and CD11b+Ly6G+Ly6Clow G-MDSC subsets via flow cytometry. Arg-1 and iNOS expression levels were gated from G-MDSCs and M-MDSCs, respectively. (A) Representative flow cytometry results are shown. (B) The percentage and (C) absolute number of G-MDSCs and the percentage and (E) absolute number of M-MDSCs in each group are shown. (F) Arg-1 and iNOS expression levels in G-MDSCs and M-MDSCs are shown. (G) MFI of Arg-1 in G-MDSCs, (H) MFI of iNOS in G-MDSCs, (I) MFI of iNOS in M-MDSCs, and (J) MFI of iNOS in M-MDSCs in each group are shown. *P < 0.05, **P < 0.01, ***P < 0.001. (K) Histological analysis of the lung section from mice via H&E staining (20× magnification). Images are representatives of two independent experiments (n = 6 mice per group).

Fig 5: MDSC subset distribution in splenocytes and lungs of mice. The splenocytes and the single-cell suspensions of lungs in mice were tested for CD11b+Ly6G-Ly6Chigh M-MDSC and CD11b+Ly6G+Ly6Clow G-MDSC subsets by flow cytometry. (A) Representative flow cytometry results are shown in splenocytes of mice. (B) G-MDSC percentage, (C) Absolute number of G-MDSCs, (D) M-MDSC percentage and (E) Absolute number of M-MDSCs in splenocytes of each group are shown. (F) Representative flow cytometry results are shown in lungs of mice. (G) G-MDSC percentage, (H) Absolute number of G-MDSCs, (I) M-MDSC percentage and (J) Absolute number of M-MDSCs in lungs of each group are shown. The values are mean±SEM of 12 mice from two independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001.