Fig 1: Chronic Inflammation Induces IL-10+ MDSC Accumulation in Colon Lamina Propria(A) RNA was isolated from colon tissues from mice treated with DSS at days 0, 7,14,21, and 28 and analyzed for IL-10 and CD11b mRNA expression levels by RT-PCR.(B) RNA was isolated from colon tissues from tumor-free control mice (n = 3) and from tumor tissues of AOM/DSS-treated mice (n = 3) and analyzed for the expression levels of IL-10 and CD11b by semiquantitative RT-PCR (left) and q-RT-PCR (right) using ß-actin as an internal control. Each column represents the mean of triplicates of one mouse. Bar, SD.(C) CD11b+Gr1+ MDSC profiles in the spleen cells of mice at days 0, 7, 14, 21, and 28 after DSS treatment, as in (A). Shown are representative results from one of three mice.(D) Quantitation of CD11b+Gr1+, as shown in (C). Bar, SD.(E) CD11b+Gr1+ MDSC levels in the lamina propria of colon tissues from control mice (n = 3) and DSS-treated mice (n = 3). Shown are representative results. The percentage of CD11b+ Gr1+ cells was quantified (right). Bar, SD.(F) Spleen cells were stained with CD11b- and Gr1-specific antibodies, followed by intracellular staining with IL-10-specific antibody. The IL-10+ cells were then gated and analyzed for CD11b+Gr1+ cells. Shown are representative results from one of three mice.(G) Quantification of the percentage of IL10+ spleen cells (left) and the percentage of CD11b+Gr1+ in the IL-10+ spleen cells (right). Bar, SD.(H) Lamina propria cells were extracted from colon tissues and stained with CD11b- and Gr1-specific antibodies, followed by intracellular staining with IL-10-specific antibody. The IL-10+ cells were then gated and analyzed for CD11b+Gr1+ cells. Shown are representative results from one of three mice.(I) Quantification of the percentage of IL-10+ lamina propria cells (left) and the percentage of CD11b+Gr1+ in the IL-10+ lamina propria cells (right). Bar, SD.
Fig 2: Streptococcus pneumoniae clearance is improved by IL-10R–deficient neutrophils. (A) Lung burden of S. pneumoniae detected at 72 h postinfection with 106 CFU/mouse intratracheally (i.t.) in WT or Il10−/− mice treated with isotype control (−) or anti-Ly6G antibodies (+) intraperitoneally 24 h prior to S. pneumoniae infection (n = 9 mice/group). (B) Schematic of neutrophil adoptive transfer. (C, D) Donor neutrophils labeled with CFSE detected by flow cytometry (C) and lung burden of S. pneumoniae (D) in recipient WT mice 72 h postinfection with S. pneumoniae 106 CFU/mouse i.t., with adoptive transfer of neutrophils purified from WT or LysMcrexIL-10Rflox mice at 24 h postinfection (n = 9 recipient mice/group). Data are pooled from 3 independent experiments and are displayed as mean ± SEM. **P < 0.01; Kruskal-Wallis with Dunn's post hoc test (A) or Mann-Whitney U test (C-D).
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