HSP47+myofibroblasts are accumulated in the fibrotic skin lesion of chronic GVHD. BALB/c mice were transplanted with 8 × 106 BM cells plus 2.5 × 107 splenocytes from allogeneic B10.D2 or syngeneic BALB/c donors on day 0, following 6 Gy of TBI. Skin samples were harvested on day +42 after BMT for histological analyses. (A) H&E and MT images. (B) Immunofluorescent images for HSP47 (green) and α-SMA (red) with DAPI (blue) counterstaining. The area in the white rectangles is magnified and is shown below the original images. (C) Immunofluorescent images for RBP1 (green) and α-SMA (red) with DAPI (blue) counterstaining. (D-F) A group of mice were intraperitoneally injected with 0.5 mg/body of αCSF1R 3 times per week after BMT, and skin samples were harvested on day +42. (D) Immunofluorescent images for HSP47 (green) and F4/80 (red) with DAPI (blue) counterstaining. The area in the white rectangles is magnified and is shown below the original images. (E) Immunofluorescent images for TGF-β (red) with DAPI (blue) counterstaining. (F) Positive pixels of F4/80 (left) and HSP47 (right) on skin samples from allogeneic and syngeneic controls and αCSF1R-treated allogeneic mice (n = 7-10/group). Data from 2 independent experiments were combined and are shown as means ± standard error of the mean (SEM). Magnification, ×20. Scale bar, 50 μm. *P < .05; **P < .01. Allo, allogeneic; Syn, syngeneic.
