Pirfenidone treatment shows variable efficacy in the scleroderma models. (A-D) BALB/c mice were conditioned with TBI (7 Gy, day 0), followed by infusion of 107 B10.D2 BM only or plus 1.8 × 106 CD4 and 0.9 × 106 CD8 T cells (day 0). In the treatment group, mice were treated with pirfenidone from day 21 to 55. (A) Skin clinical score based on the area of skin lesion. (B) Hematoxylin and eosin (HE) staining of the skin (left) and pathology score (right). Pirfenidone reduced both clinical score and pathology score. (C) Trichrome staining of skin. Collagen was stained blue. Fibrosis was scored on a 1 to 5 scale based on the intensity of blue staining. Pirfenidone significantly reduced collagen deposition in skin. (D) Immunofluorescence staining of macrophage in skin samples. CD68 (red) identified macrophages. Pirfenidone significantly reduced macrophage infiltration in skin. (E) Flow cytometry analysis cytokine production and Treg percentage of splenocytes from transplanted mice. (F) B6D2F1 mice were conditioned with TBI (11 Gy split into 2 doses, day 1) followed by infusion of 5 × 106 B6 BM only or plus 1 × 106 purified T cells (day 0). Where indicated, mice were treated with pirfenidone from days14 to 28 or days 14 to 41. HE staining on days 28 (top) and 42 (bottom) and day28 F4/80 staining (middle) indicating that pirfenidone alleviated macrophage infiltration but failed to significantly improve pathologic changes. In each experiment, 5 to 10 mice from each group were tested. *P ≤ .05, **P < .01, ***P < .001; data are shown as the mean ± SEM.
