Figure 4. Allogeneic T cell interaction with colonic epithelial cells and AREG blockade in mice and human. (A) sST2 concentration in plasma collected every 5 d after HCT from the BALB/c mice receiving 5 × 106 BM cells plus 106 from (n = 4, from two independent experiments, unpaired t test; data are shown as mean ± SEM; *, P < 0.05; **, P < 0.01). (B) TNF and IFN-γ concentrations in plasma collected every 5 d after HCT from BALB/c mice receiving 5 × 106 BM cells plus 1 × 106 from syngeneic T cells (BALB/c mice) or T1, T9, or T9IL-33 T cells from B6 WT mice (n = 4, from two independent experiments, unpaired t test; data are shown as mean ± SEM; *, P < 0.05, **. P < 0.01). (C) Ratio of AREG/actin mRNA expression in WT or IL-9−/− T9IL-33 cells. (D) Ratio of mST2/actin expression (left) and sST2/actin expression (right) in WT or IL-9−/− T9IL-33 cells (n = 3, from three independent experiments, unpaired t test; data are shown as mean ± SEM; *, P < 0.05; **, P < 0.01). (E) Ex vivo expression of IFN-γ and IL-17 in gut CD4 T cells collected on day 14 after allo-HCT with allogeneic T1, WT T9IL-33, or ST2−/− T9IL-33 cells (n = 4, from two independent experiments, unpaired t test; data are shown as mean ± SEM; **, P < 0.01; ***, P < 0.001). (F) Ex vivo expression of IFN-γ and IL-17 in gut CD4 T cells collected on day 14 after allo-HCT with T9IL-33 cells from Foxp3-GFP reporter mice depleted or nondepleted of Foxp3 T reg cells (n = 3, unpaired t test; data are shown as mean ± SEM). (G) AREG expression on in vitro differentiated human T1, T9, and T9IL-33 CD4 cells from healthy donors (n = 3, from three independent experiments, unpaired t test; data are shown as mean ± SEM; *, P < 0.05; **, P < 0.01; ***, P < 0.001). (H) Percentage of residual live human colonic cells after co-culture with T1, T9, T9IL-33 cells for 6 h in the presence of anti-AREG blocking antibody or isotype control and without T cells as control (n = 3, from three independent experiments, unpaired t test; data are shown as mean ± SEM; **, P < 0.01; ***, P < 0.001).
Figure 4.

Allogeneic T cell interaction with colonic epithelial cells and AREG blockade in mice and human. (A) sST2 concentration in plasma collected every 5 d after HCT from the BALB/c mice receiving 5 × 106 BM cells plus 106 from (n = 4, from two independent experiments, unpaired t test; data are shown as mean ± SEM; *, P < 0.05; **, P < 0.01). (B) TNF and IFN-γ concentrations in plasma collected every 5 d after HCT from BALB/c mice receiving 5 × 106 BM cells plus 1 × 106 from syngeneic T cells (BALB/c mice) or T1, T9, or T9IL-33 T cells from B6 WT mice (n = 4, from two independent experiments, unpaired t test; data are shown as mean ± SEM; *, P < 0.05, **. P < 0.01). (C) Ratio of AREG/actin mRNA expression in WT or IL-9−/− T9IL-33 cells. (D) Ratio of mST2/actin expression (left) and sST2/actin expression (right) in WT or IL-9−/− T9IL-33 cells (n = 3, from three independent experiments, unpaired t test; data are shown as mean ± SEM; *, P < 0.05; **, P < 0.01). (E) Ex vivo expression of IFN-γ and IL-17 in gut CD4 T cells collected on day 14 after allo-HCT with allogeneic T1, WT T9IL-33, or ST2−/− T9IL-33 cells (n = 4, from two independent experiments, unpaired t test; data are shown as mean ± SEM; **, P < 0.01; ***, P < 0.001). (F) Ex vivo expression of IFN-γ and IL-17 in gut CD4 T cells collected on day 14 after allo-HCT with T9IL-33 cells from Foxp3-GFP reporter mice depleted or nondepleted of Foxp3 T reg cells (n = 3, unpaired t test; data are shown as mean ± SEM). (G) AREG expression on in vitro differentiated human T1, T9, and T9IL-33 CD4 cells from healthy donors (n = 3, from three independent experiments, unpaired t test; data are shown as mean ± SEM; *, P < 0.05; **, P < 0.01; ***, P < 0.001). (H) Percentage of residual live human colonic cells after co-culture with T1, T9, T9IL-33 cells for 6 h in the presence of anti-AREG blocking antibody or isotype control and without T cells as control (n = 3, from three independent experiments, unpaired t test; data are shown as mean ± SEM; **, P < 0.01; ***, P < 0.001).

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