Marilen Benner

THREE TYPES OF TREG CONTROL PLACENTAL INFLAMMATION 57 3 Figure 2. Decidual Tregs suppress proliferation of Teffs. (A) CFSE dilution of 6-12-week decidual CD4+ Teffs (upper panel) and CD8+ Teffs (lower panel) stimulated with anti-CD3/28 and co-cultured with or without CD25HI, PD1HI, or TIGIT+ Tregs during 4 days in a 1:2, Treg: Teff ratio. Percentages of the undivided Teffs are shown. (B–G) Fold change (FC) in the percentage of undivided cells (B–D) and in the division index ( E–G) after the addition of CD25 HI , PD1 HI , or TIGIT+ Tregs, compared to CD4+ Teffs 6–12 weeks (B and E) , CD8+ Teffs 6–12 weeks (C and F) , and CD4+ Teffs >37 weeks ( D and G) cultured alone. (H) Addition of IL-10R blocking antibodies to Teffs +PD1 HI but not Teffs + CD25 HI cultures inhibit suppression of proliferation, measured by FC in percentage of undivided cells, compared to isotype control cultures. Bars represent median and interquartile range; In (B) – (G) : n = 7–11; (H): n = 3–4; *p < 0.05, **p < 0.01 and ***p < 0.001; values > 1 for FC % undivided cells and < 1 for FC division index represent suppression of proliferation. Division index reflects the number of divisions per cell as calculated in FlowJO v7.6.5. See also Figure S4.