Figure 1

Close localization of MDSCs and B cells in the spleen. Tumors were induced in C57BL/6 wild-type mice by subcutaneously injecting MCA205 (5 × 10⁵) tumor cells. Then, the spleens were studied after three weeks. Untreated mice served as controls. *P<0.05 and **P<0.01, as determined with Student’s t-test. (a) Spleen sections were stained for B220 (green) and CD11b (red) and were assessed by cLSM. The cLSM images are representative of three independent experiments with 3 mice per group; Scale bars, 100 μm. (b) Isolated splenocytes were stained for CD11b and Gr1, and the CD11b⁺Gr1⁺ cells were analyzed by flow cytometry. The percentages are indicated in the representative dot plots. The results are shown as the mean values±s.e.m. of the CD11b⁺Gr1⁺ cell percentages from three independent experiments with 3 mice per group per experiment. (c) Spleen sections were stained for B220 (green) and IgA (red) and they were subsequently analyzed by fluorescence microscopy. The left panel shows representative images (scale bars, 200 μm) and the right panel indicates the mean values±s.e.m. of the B220⁻IgA⁺ cell percentages from three independent experiments with 2-3 mice per group. (d) Isolated splenocytes were stained for B220 and IgA, and the B220⁻IgA⁺ cells were analyzed by flow cytometry. The percentages are indicated in the representative dot plots. The results are shown as the mean values±s.e.m. of the B220⁻IgA⁺ cell percentages from three independent experiments with 3 mice per group per experiment. (e) Spleen sections that were stained for Gr1 (green) and IgA (red) were analyzed by fluorescence microscopy. The images are representative of two independent experiments with 3 mice per group; scale bars, 50 μm. cLSM, confocal laser scanning microscope; IgA, immunoglobulin A; MDSCs, myeloid-derived suppressor cells.