Fig 2: IHC staining for immune cells populations.a-g. IHC (yellow) for CD4 (a), CD8 (b), CD68 (c), CD14 (d), CD20 (e), CD66b (f) and Leishmania OPB (g). Left and right images correspond to boxed areas of images shown in Fig. 4a right. Images are for a single participant (LC001) and pair wise staining was performed on serial sections. Scale bar in all images represents 0.2 mm.

Fig 3: Anti-DCIR mAb promotes neutrophil clearance through ADCP and ADCC.(A) LPS primed human neutrophils were labeled CellTracker Green. Neutrophils were incubated with 5 μg/mL anti-DCIR mAbs (3A4, 9D9, 5E11, 3F7) or isotype for 30 minutes on ice. Neutrophils and monocyte-derived macrophages were cocultured (ratio 4:1). (B) ADCP were quantitated by flow cytometry detecting CellTracker Green+CD11b+CD66b–HLA-DR+macrophage after 2 hours coculture with 3A4 (n = 12), 9D9 (n = 6), 5E11 (n = 6), 3F7(n = 12), or isotype (n = 11). Data are normalized to 3A4 group. (C) LPS primed neutrophils were incubated with 3A4, 9D9, 5E11, 3F7, or isotype for 30 minutes on ice. Neutrophils and Jurkat cells containing an ADCC reporter were cocultured (ratio 4:1). (D) ADCC were quantitated by luminescence overnight. Representative data from 2 studies are shown. (E) Confocal flow cytometry of human monocytes treated with 10 μg/mL A647-labeled antibodies (red) for 0, 15, and 30 minutes at 37°C. Antibody internalization was determined by colocalization (yellow) of A647-labeled antibodies in lysosome, marked by A488 anti-LAMP1 antibody (green). (F and G) HuDCIR-KI mice received i.p. administration of 10 mpk 5E11 with WT (n = 25) or LALA mutant (n = 7) huIgG1 Fc or isotype (n = 25) 1 day before the i.p. injection of 0.5 mL 2 mg/mL ZymD. Relative leukocytes and neutrophils induction in peritoneal lavage were quantitated 6 hours after the ZymD injection. Data are normalized to isotype group. (H–K) Relative leukocytes and neutrophils infiltration (H and I) and cytokine in peritoneal fluid (J and K) collected 6 hours after ZymD injection, from mice pretreated with 5E11 with mouse IgG2b (5E11-mIgG2b) (n = 5), Fc-matched isotype (iso-mIgG2b) (n = 5), or anti–Gr-1 mAb (RB6-8C5) (n = 5) 1 day before the peritonitis. Data are normalized to isotype group. Each dot represents 1 biological replicate. Means ± SEM are shown, and statistical significance is determined by 1-way ANOVA with Dunnett’s correction compared with the isotype condition. *P < 0.05, **P < 0.01, ***P < 0.001.