Figure 1.
Figure 1. Hair follicles and rete-like prominences in the lingual epithelium are damaged in GVHD. Lethally irradiated B6 mice were transplanted with 20 × 106 G-CSF–mobilized splenocytes from syngeneic (Syn) B6 or allogeneic (Allo) BALB/c mice on day 0, except for (G,P) using B6-Lck-Cre × R26tdTomato and for (K) using B6-Il17aCre × R26eYFP as donors. (A) Clinical GVHD scores from 3 experiments (n = 11 per group). (B) Survival curves from 6 independent experiments (n = 20 per group). (C) Serum levels of IFN-γ on day +14 after SCT from 3 independent experiments (n = 8-10 per group). (D-F) The back skins were harvested on day +14 (D) and +28 (E-F) after SCT. The representative images of hematoxylin and eosin (H&E) staining (D-E) and pathological GVHD scores (F) from 3 independent experiments. The arrowhead indicates dermal epidermal detachment. (G) Immunofluorescent images of tdTomato+ donor T cells (red) with 4′,6-diamidino-2-phenylindole (DAPI) nuclear staining (blue) in the skin harvested on day +10 after SCT. Dashed line shows the epidermal-dermal junction. (H-L) Flow cytometric analysis of skin-infiltrating cells on day +14 after SCT. The absolute numbers of donor T cells (H), CD11b+ myeloid cells (I), IFN-γ+ T cells (J), IL-17 eYFP+ T cells (K), and mean fluorescence intensity (MFI) of CXCR3 on T cells (L) from 2 experiments (n = 8-10 per group). (M-N) Total RNA was extracted from the whole skin (M) or purified CD45+ cells and CD45− cells from the back skin (N) on day +14 after SCT and the expression of Cxcl9 was evaluated by Q-PCR (n = 8-10 per group from 2 independent experiments). H&E staining (O) and immunofluorescent staining of tdTomato+ donor T cells (red) with DAPI (blue) staining (P) of lingual samples harvested on day +7 after SCT. Dashed line shows the junction of epithelium and lamina propria. Original magnification ×20 (D,G,O,P) and ×10 (E). Scale bar, 50 μm. The Mann-Whitney U test or 1-way ANOVA followed by the Tukey posttest was used to compare the data (*P < .05; **P < .01; ***P < .005). Data represent the mean ± SEM. rRNA, ribosomal RNA.

Hair follicles and rete-like prominences in the lingual epithelium are damaged in GVHD. Lethally irradiated B6 mice were transplanted with 20 × 106 G-CSF–mobilized splenocytes from syngeneic (Syn) B6 or allogeneic (Allo) BALB/c mice on day 0, except for (G,P) using B6-Lck-Cre × R26tdTomato and for (K) using B6-Il17aCre × R26eYFP as donors. (A) Clinical GVHD scores from 3 experiments (n = 11 per group). (B) Survival curves from 6 independent experiments (n = 20 per group). (C) Serum levels of IFN-γ on day +14 after SCT from 3 independent experiments (n = 8-10 per group). (D-F) The back skins were harvested on day +14 (D) and +28 (E-F) after SCT. The representative images of hematoxylin and eosin (H&E) staining (D-E) and pathological GVHD scores (F) from 3 independent experiments. The arrowhead indicates dermal epidermal detachment. (G) Immunofluorescent images of tdTomato+ donor T cells (red) with 4′,6-diamidino-2-phenylindole (DAPI) nuclear staining (blue) in the skin harvested on day +10 after SCT. Dashed line shows the epidermal-dermal junction. (H-L) Flow cytometric analysis of skin-infiltrating cells on day +14 after SCT. The absolute numbers of donor T cells (H), CD11b+ myeloid cells (I), IFN-γ+ T cells (J), IL-17 eYFP+ T cells (K), and mean fluorescence intensity (MFI) of CXCR3 on T cells (L) from 2 experiments (n = 8-10 per group). (M-N) Total RNA was extracted from the whole skin (M) or purified CD45+ cells and CD45− cells from the back skin (N) on day +14 after SCT and the expression of Cxcl9 was evaluated by Q-PCR (n = 8-10 per group from 2 independent experiments). H&E staining (O) and immunofluorescent staining of tdTomato+ donor T cells (red) with DAPI (blue) staining (P) of lingual samples harvested on day +7 after SCT. Dashed line shows the junction of epithelium and lamina propria. Original magnification ×20 (D,G,O,P) and ×10 (E). Scale bar, 50 μm. The Mann-Whitney U test or 1-way ANOVA followed by the Tukey posttest was used to compare the data (*P < .05; **P < .01; ***P < .005). Data represent the mean ± SEM. rRNA, ribosomal RNA.

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