Fig. 5

Peritoneal exudate cells (PEC) isolated from Mesocestoides vogae-infected mice possess suppressive capacities and can be subdivided into two subsets. Ly6C+ and Ly6G+ cells were isolated using MACS from the peritoneal cell population at day 35 post-infection and cultured with CFSE-labeled T cells stimulated with anti-CD3/ anti-CD28 mAbs for 72 h at different ratios. a Flow cytometry plots of PEC from healthy and infected mice showing the expression of CD11b and Gr-1 before and after MACS sort. b Representative histograms showing suppression of spleen T cells by sorted Ly6C+ or Ly6G+ in vitro (1:4 ratio MDSC to T cells). c The graph represents the proliferation inhibition of CD3/CD28-activated spleen T cells (the ratio of myeloid cell subsets to T cells as indicated in the graph). Results represent means ± SD of triplicates. Statistical significance was analyzed using Student's t test, and significantly different values are indicated as: **p < 0.01, ***p < 0.001. d The sorted cells were subjected to May-Grünwald/Giemsa staining. Representative pictures showing the morphology of sorted subsets are depicted. Original magnification, ×1000, scale bar = 20 μm. e mRNA expression of iNOS and Arg-1 in PEC of healthy mice (control) and sorted Ly6C+ or Ly6G+ cells. Statistical significance was analyzed using two-way ANOVA followed by Bonferroni's multiple comparison tests and significantly different values are indicated as: ***p < 0.001. f Immunofluorescent assay of iNOS and Arg-1 in whole peritoneal cells. Hoechst staining was used to visualize nuclei. Original magnification, ×1000, scale bar = 10 μm.