Figure 2. Impact of T1 versus T9IL-33 cells on gut pathology and effect of ST2–IL-33 signaling on gut T cell proliferation, viability, and migration, T reg expansion, and ILC2 expansion. (A) Pathology index of mouse intestines at day 10 after allo-HCT with either T1 or T9IL-33 cells (n = 3, from three independent experiments, unpaired t test; data are shown as mean ± SEM; *, P < 0.05). Specimens were harvested, placed in 10% buffered formalin, embedded in paraffin, cut into 5-µm-thick sections, and stained with hematoxylin and eosin for histological examination. Slides were coded without reference to mouse type or prior treatment status and examined systematically by a single pathologist. (B) Representative plots of CFSE-labeled in gut T cells collected from mice on day 5 after all-HCT with syngeneic BALB/c T cells or allogeneic in vitro differentiated T cells. (C) Absolute counts of gut-infiltrating T cells from mice on day 10 after allo-HCT with syngeneic BALB/c T cells or allogeneic in vitro differentiated T cells (n = 3, mean ± SEM, from three independent experiments, unpaired t test; data are shown as mean ± SEM). (D) Representative plots of α4β7 and CRK (v-crk sarcoma virus CT10 oncogene homologue [avian], also called p38) and CCR5 in CD4 T cells infiltrating the gut at day 10 after HTC (n = 3, unpaired t test; data are shown as mean ± SEM). (E) Representative plots of Annexin V and fixable viability dye (FVD) staining of gut T cells at day 10 post-HTC (n = 3). (F) Representative plots of CD4 and FoxP3 and a bar graph showing the frequency of T reg cells (CD4+FoxP3+) in gut-infiltrating CD4 T cells at day 10 after HTC (n = 4, unpaired t test; data are shown as mean ± SEM). (G) Representative plots of mST2 and Gata3 in Lin−CD45+CD90.2+ cells (ILC2 markers, n = 4, unpaired t test; data are shown as mean ± SEM). (H) Ex vivo expression of Foxp3-GFP in gut CD4 T cells collected on day 14 after allo-HCT with either Foxp3-GFP–depleted or nondepleted allogeneic T9IL-33 cells (n = 3, mean ± SEM). (I) Clinical scores of GVHD and survival curves for C3H.SW mice receiving B6 BM cells and in vitro differentiated Foxp3-GFP–depleted or nondepleted T9IL-33 cells by flow cytometry (n = 9 per group, unpaired t test; data are shown as mean ± SEM).
Figure 2.

Impact of T1 versus T9IL-33 cells on gut pathology and effect of ST2–IL-33 signaling on gut T cell proliferation, viability, and migration, T reg expansion, and ILC2 expansion. (A) Pathology index of mouse intestines at day 10 after allo-HCT with either T1 or T9IL-33 cells (n = 3, from three independent experiments, unpaired t test; data are shown as mean ± SEM; *, P < 0.05). Specimens were harvested, placed in 10% buffered formalin, embedded in paraffin, cut into 5-µm-thick sections, and stained with hematoxylin and eosin for histological examination. Slides were coded without reference to mouse type or prior treatment status and examined systematically by a single pathologist. (B) Representative plots of CFSE-labeled in gut T cells collected from mice on day 5 after all-HCT with syngeneic BALB/c T cells or allogeneic in vitro differentiated T cells. (C) Absolute counts of gut-infiltrating T cells from mice on day 10 after allo-HCT with syngeneic BALB/c T cells or allogeneic in vitro differentiated T cells (n = 3, mean ± SEM, from three independent experiments, unpaired t test; data are shown as mean ± SEM). (D) Representative plots of α4β7 and CRK (v-crk sarcoma virus CT10 oncogene homologue [avian], also called p38) and CCR5 in CD4 T cells infiltrating the gut at day 10 after HTC (n = 3, unpaired t test; data are shown as mean ± SEM). (E) Representative plots of Annexin V and fixable viability dye (FVD) staining of gut T cells at day 10 post-HTC (n = 3). (F) Representative plots of CD4 and FoxP3 and a bar graph showing the frequency of T reg cells (CD4+FoxP3+) in gut-infiltrating CD4 T cells at day 10 after HTC (n = 4, unpaired t test; data are shown as mean ± SEM). (G) Representative plots of mST2 and Gata3 in Lin−CD45+CD90.2+ cells (ILC2 markers, n = 4, unpaired t test; data are shown as mean ± SEM). (H) Ex vivo expression of Foxp3-GFP in gut CD4 T cells collected on day 14 after allo-HCT with either Foxp3-GFP–depleted or nondepleted allogeneic T9IL-33 cells (n = 3, mean ± SEM). (I) Clinical scores of GVHD and survival curves for C3H.SW mice receiving B6 BM cells and in vitro differentiated Foxp3-GFP–depleted or nondepleted T9IL-33 cells by flow cytometry (n = 9 per group, unpaired t test; data are shown as mean ± SEM).

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