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. 2017 Jun 8;152(1):89–101. doi: 10.1111/imm.12751

Figure 3.

Figure 3

Cord blood myeloid‐derived suppressor cells (CB‐MDSC) induce T helper type 2 (Th2) cells via ArgI and reactive oxygen species (ROS) and partially dependent on monocytes. CB‐MDSC were enriched from cord blood mononuclear cells (CBMC) and added to peripheral blood mononuclear cells (PBMC), CD3+ T cells or CD14‐depleted PBMC isolated from healthy adult controls either with direct cell/cell‐contact or in transwells. After 5 days of culture, surface staining was performed and cells were analysed by flow cytometry. (a) Scatter plot with bars shows the percentage of CD4+ CXCR3− CCR4+ CCR6− Th2 cells without addition of CB‐MDSC (white bar), with addition of CB‐MDSC in direct cell/cell‐contact (grey bar) or separated by transwells (checked bar). Bars represent pooled data from 11 independent experiments and each point represents an individual sample. **P = 0·008; paired t‐test. (b, c) Scatter plots with bars shows induction of Th2 cells by CB‐MDSC alone (white bar) or under addition of inhibitors for ArgI (nor‐NOHA), ROS (catalase), iNOS (L‐NMMA) and IDO (1‐MT) in ascending grey scales. (b) Percentages of Th2 cells without addition of CB‐MDSC (blank bars) and with addition of CB‐MDSC (checked bars). n = 5 or n = 6; **P < 0·01, *P < 0·05, ns = not significant; Wilcoxon matched‐pairs signed rank test. (c) Induction of Th2 cells was determined as ratio between percentages of Th2 cells with addition of CB‐MDSC and percentages of Th2 cells without addition of CB‐MDSC. n = 5 or n = 6, *P < 0·05, ns = not significant in comparison to without (w/o) inhibitors; Wilcoxon matched‐pairs signed rank test. (d) Scatter plot with bars show induction of Th2 cells by CB‐MDSC alone (white bar) or under addition of nor‐NOHA (light grey bar) or nor‐NOHA and catalase in combination (checked bar). n = 5, *P < 0·05, Wilcoxon matched‐pairs signed rank test. (e) Scatter plot with bars show the percentage of CD4+ CXCR3− CCR4+ CCR6− Th2 cells after culture of CD3+ T cells without (white bar) or with CB‐MDSC (grey bar) and with CB‐MDSC and CD14+ monocytes (checked bar) or with CB‐MDSC and CD19+ B cells (striped bar). n = 6, *P = 0·02, Wilcoxon matched‐pairs signed rank test. (f) Scatter plot with bars shows the percentage of CD4+ CXCR3− CCR4+ CCR6− Th2 cells after culture of PBMC (filled bars) or CD14‐depleted PBMC (checked bars) without addition of CB‐MDSC (white bars) or with addition of CB‐MDSC (grey bars). n = 8, **P = 0·003; Wilcoxon matched‐pairs signed rank test.