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. Author manuscript; available in PMC: 2016 Nov 12.
Published in final edited form as: Cell Rep. 2016 Oct 25;17(5):1318–1329. doi: 10.1016/j.celrep.2016.09.082

Figure 2. ITE induces functional human FOXP3+ Tregs in the presence of TGFβ1.

Figure 2

(A) Suppressive activity of human naïve CD4+ T cells activated with αCD3/αCD28 and TGFβ1 in the presence of CTL, ITE, or TCDD. Percent suppression is depicted as the mean ± SEM pooled from at least 2 independent experiments with 10 unique donors.

(B) qPCR analysis on RNA isolated from T cells stimulated with αCD3/αCD28 and IL2 in the presence of CTL, TGFβ1, TGFβ1 + ITE, or TGFβ1 + TCDD. Target fold change was calculated using vehicle control with the dashed line representing the normalized control value of 1 with relative expression shown as the pooled mean ± SEM (n ≥ 10).

(C) Representative flow cytometric analysis of TBX21, GATA3, and RORC protein expression in T cells following activation using αCD3/αCD28 and IL2 in the presence of CTL, TGFβ1, or TGFβ1 + ITE from 3 independent experiments.

(D) qPCR analysis on RNA isolated from T cells stimulated with αCD3/αCD28 in the presence of CTL, TGFβ1, TGFβ1 + ITE, or TGFβ1 + TCDD. Target fold change was calculated using vehicle control with the dashed line representing the normalized control value of 1 with relative expression shown as the pooled mean ± SEM (n ≥ 7). ‡ P < 0.05 compared to control.

(E) Representative flow cytometric analysis of FOXP3, IL-10, CD39, and GZMB protein expression in T cells following activation using αCD3/αCD28 and IL2 in the presence of CTL, TGFβ1, or TGFβ1 + ITE from 2 independent experiments.

(F) Effect of CD39 blockade using CD39 blocking antibodies on the suppressive activity of TGF+ITE (left panel) or TGF+TCDD (right panel) with the mean depicted ± SEM (n ≥ 12). * P < 0.05, ** P < 0.01, *** P < 0.001.