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. 2015 Jan 14;83(2):743–758. doi: 10.1128/IAI.02639-14

FIG 7.

FIG 7

B cell-derived IL-10 in conjunction with B cell-mediated induction of IL-10 in accessory cells protects on-demand hematopoiesis during responses to Pneumocystis lung infection. (A) Total bone marrow cell counts from IFrag−/− mice that were either left untransferred or that had received B cells isolated from wild-type or IL-10−/− donor mice on days 0, 10, and 16 after Pneumocystis lung infection. (B) Comparative analysis of hematopoietic progenitor activity for the granulocytic/monocytic lineage were performed by plating 1 × 105 total bone marrow cells per mouse of each comparison group in hematopoietic colony-forming assays (CFU assays) using reagents from StemCell Technologies. All samples were plated in duplicate. Total CFU numbers were plotted. (C) Serum IL-10 levels were measured in each group in the course of infection (from days 0 to 16 after infection) using ELISA reagents from R&D Systems. (D) Freshly flushed bone marrow cells were spun onto glass slides and evaluated morphologically by light microscopy following Diff-Quik staining. Bone marrow cells from uninfected and untransferred IFrag−/− mice are shown compared to IFrag−/− mice with Pneumocystis lung infections that had received B cells from either wild-type or IL-10−/− mice at 16 days (d.16) or were left untransferred. (E) Shown are the percentage of mature (band and segmented) neutrophils identified via morphological analysis and confirmed by FACS analysis in the comparison groups over the course of Pneumocystis lung infection. (F) Pneumocystis lung burden displayed as the number of nuclei in log10 units between the three groups over the 16-day course of infection. For statistical analysis of data between the two groups over time, a two-way ANOVA was performed followed by a Bonferroni posthoc test. Values that are significantly different are indicated as follows: * or #, P < 0.05; **, P < 0.01; *** or ###, P < 0.001. Each B cell-transferred group was compared separately to untransferred IFrag−/− group. Values compared to wild-type-B cell-transferred mice are shown as asterisks, and values compared to IL-10−/− B cell-transferred mice are shown as number or pound signs.