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. 2021 Sep 1;12:716606. doi: 10.3389/fimmu.2021.716606

Figure 3.

Figure 3

MAPC expanded Tregs exhibit superior suppression of antigen specific responses. (A–C) Suppression assay using 3rd party PBMC stimulated with CD3/CD28 microbeads for 6 days in the presence or absence of Treg or MulTreg lines. (A). Flow cytometry plots showing proliferation in responder T cells from PBMC stimulated with CD3/CD28 microbeads (1:10 bead to cell) in the presence or absence of autologous, expanded Treg or MulTreg lines at a ratio of 1:2 Treg : PBMC. (B) Bar graph displaying % suppression of responder T cell proliferation at different ratios of Treg or MulTreg to PBMC. (C) Bar graph displaying cytokine levels in tissue culture supernatant from suppression assays in panel (B) (ratio 1:2 Treg : PBMC). n = 5. (D–F) Suppression assay using autologous PBMC stimulated with Flu-HA for 6 days in the presence or absence of Treg or MulTreg lines. (D) Flow cytometry plots showing proliferation in responder CD4 T cells from PBMC stimulated with Flu-HA in the presence or absence of autologous, expanded Treg or MulTreg lines at a ratio of 1:5 Treg : PBMC. (E) Bar graph displaying % suppression of responder CD4+ T cell proliferation at different ratios of Treg : PBMC, CD4 (left) and CD8 (right). (F) Bar graph displaying cytokine levels in tissue culture supernatant from suppression assays in panel (E) (ratio of 1:5). Error bars represent the SEM of 5 donors. Stats from Wilcoxon matched-pairs analysis (B, E) or Friedman test (C, F). *p < 0.05, **p < 0.01, ns, not signficant.