Evidence of cGvHD and antiviral immune response in recipients with untreated splenocytes. Irradiated CB6F1 recipients of 3 × 106 untreated donor splenocytes (cGvHD), 10 × 106 amotosalen-treated donor splenocytes (no-GvHD–treated splenocyte recipients), or TCD BM alone (no GvHD) were infected with MCMV (5 × 104 PFU intraperitoneally) on 100+ days after transplantation. (A) Percentage weight loss was calculated from the initial weight measured on day 0 of transplantation. Here, day 0 represents the percentage weight loss measured on day 133 after transplantation (□, cGvHD, splenocytes recipients; ▪, no GvHD; and ▴, TCD BM). (B) Recipients with cGvHD, no-GvHD–treated splenocyte recipients, and TCD BM were killed on day 100 after transplantation (day 0 after MCMV infection) and day 10 after 5 × 104 PFU MCMV infection. Skin, liver, and small intestine were collected in formalin solution and cGvHD scores were assessed from the histologic tissue sections of skin, liver, and small intestines. The average mean value and standard deviation of cGvHD scores obtained from the 3 organs of recipients with cGvHD (□), no GvHD (▪), and TCD BM (▴) on days 0 and 10 after MCMV infection. (C) The presence of sclerodermoid sclerosis and fibrosis (yellow arrows) is shown in the histologic slides of skin tissue sections collected from the recipients with cGvHD, no GvHD, and TCD BM under uninfected conditions (day 100 after transplantation) and day 10 after MCMV infection (day 110 after transplantation). The data are representative of 1 of 2 similar experiments and 4 to 5 mice were used per group in each time point. (D) Recipients with cGvHD, no GvHD, and TCD BM alone were infected with MCMV (5 × 104 PFU intraperitoneally) at 100 days after transplantation. Liver and spleen were aseptically collected on day 3 after infection and viral load per organs was determined by spreading the tissue homogenates on a 3T3-cell confluent monolayer as described in “Materials and methods.” Mean viral PFU and standard deviations obtained from each of the infected organs of 5 to 6 mice per group were used (*P < .001, Student t test).
