Figure 1.
Hepatic GVHD induces apoptosis and transitory proliferation of BECs. (A-G) Lethally irradiated BDF1 mice were transplanted from allogeneic B6 (Allo) or syngeneic BDF1 (Syn) donors on day 0. Clinical GVHD scores (A; n = 11-12/group) and pathologic liver GVHD scores (B; n = 10/group) are shown. (C-G) Flow cytometric analyses of liver mononuclear cells were performed at indicated time points (n = 7-12/group). See also supplemental Figures 1 and 2. Absolute numbers of donor CD4+ T cells (C), CD8+ T cells (D), donor monocytes (E), infiltrating donor macrophages (F), and Kupffer cells (G) are shown. (H,I) Lethally irradiated B6 (Syn) or BDF1 (Allo) mice were transplanted with purified T cells from B6 Lck-cre×R26Tomato combined with B6 T-cell-depleted-BM cells on day 0. (H) Immunofluorescent staining of CK19 (green), tdTomato+ donor T cells (red) with DAPI (4′,6-diamidino-2-phenylindole; blue) staining on day 21 after HCT. (I) Immunofluorescent staining of CK19 (green), F4/80 (red) with DAPI (blue) staining on day 28 after HCT. (J-S) Lethally irradiated BDF1 mice were transplanted as in Figure 1A. Immunofluorescent staining of cleaved caspase-3 (cCaspase3; green) and CK19 (red) with DAPI (blue) staining on day 21 after HCT. Representative images (J) and the proportion (K) of cCaspase 3+ BECs (n = 4-5/group). (L) Immunofluorescent staining of Mmp7 (green) and CK19 (red) with DAPI (blue) staining on day 21. (M) Relative expression of Mmp7 in the liver on day 21 (n = 5/group). (N,O) Immunofluorescent staining of CK19 (green) with DAPI (blue) staining on days 21 and 35. Representative images (N) and proportion (O) of CK19+ area were shown as fold change from naive mice (n = 5/group). (P,Q) Immunofluorescent staining of Ki-67 (green) and CK19 (red) with DAPI (blue) staining on days 21 and 35. Representative images (P) and proportions (Q) of Ki-67+ cells among CK19+ BECs were shown (n = 5/group). White arrows indicate clusters of Ki-67+ BECs. (R) Plasma levels of total bilirubin from the recipients (n = 10/group) and naïve BDF1 (n = 4) were shown. (S) Immunofluorescent staining of albumin (Alb; green) and CK19 (red) with DAPI (blue) staining 70 days after HCT. (A-G, O, Q, R) Data from 2 independent experiments were combined and shown as mean ± standard error (SE). (K,M) Data from 1 of 2 independent experiments are shown as mean ± SE. Bar, 100 μm. ∗P < .05, ∗∗P < .01, ∗∗∗P < .005.

Hepatic GVHD induces apoptosis and transitory proliferation of BECs. (A-G) Lethally irradiated BDF1 mice were transplanted from allogeneic B6 (Allo) or syngeneic BDF1 (Syn) donors on day 0. Clinical GVHD scores (A; n = 11-12/group) and pathologic liver GVHD scores (B; n = 10/group) are shown. (C-G) Flow cytometric analyses of liver mononuclear cells were performed at indicated time points (n = 7-12/group). See also supplemental Figures 1 and 2. Absolute numbers of donor CD4+ T cells (C), CD8+ T cells (D), donor monocytes (E), infiltrating donor macrophages (F), and Kupffer cells (G) are shown. (H,I) Lethally irradiated B6 (Syn) or BDF1 (Allo) mice were transplanted with purified T cells from B6 Lck-cre×R26Tomato combined with B6 T-cell-depleted-BM cells on day 0. (H) Immunofluorescent staining of CK19 (green), tdTomato+ donor T cells (red) with DAPI (4′,6-diamidino-2-phenylindole; blue) staining on day 21 after HCT. (I) Immunofluorescent staining of CK19 (green), F4/80 (red) with DAPI (blue) staining on day 28 after HCT. (J-S) Lethally irradiated BDF1 mice were transplanted as in Figure 1A. Immunofluorescent staining of cleaved caspase-3 (cCaspase3; green) and CK19 (red) with DAPI (blue) staining on day 21 after HCT. Representative images (J) and the proportion (K) of cCaspase 3+ BECs (n = 4-5/group). (L) Immunofluorescent staining of Mmp7 (green) and CK19 (red) with DAPI (blue) staining on day 21. (M) Relative expression of Mmp7 in the liver on day 21 (n = 5/group). (N,O) Immunofluorescent staining of CK19 (green) with DAPI (blue) staining on days 21 and 35. Representative images (N) and proportion (O) of CK19+ area were shown as fold change from naive mice (n = 5/group). (P,Q) Immunofluorescent staining of Ki-67 (green) and CK19 (red) with DAPI (blue) staining on days 21 and 35. Representative images (P) and proportions (Q) of Ki-67+ cells among CK19+ BECs were shown (n = 5/group). White arrows indicate clusters of Ki-67+ BECs. (R) Plasma levels of total bilirubin from the recipients (n = 10/group) and naïve BDF1 (n = 4) were shown. (S) Immunofluorescent staining of albumin (Alb; green) and CK19 (red) with DAPI (blue) staining 70 days after HCT. (A-G, O, Q, R) Data from 2 independent experiments were combined and shown as mean ± standard error (SE). (K,M) Data from 1 of 2 independent experiments are shown as mean ± SE. Bar, 100 μm. ∗P < .05, ∗∗P < .01, ∗∗∗P < .005.

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