Fig. 4

Anti-IL-39 antibody ameliorates immune dysregulation in scleroderma-like cGVHD. Irradiated BALB/c recipients were infused with 1 × 10⁷ bone marrow cells and 1 × 10⁶ splenocytes from the C57BL/6 mice. Fourteen days after transplantation, each mouse in the antibody group received 100 μl (100 μg) of IL-39 antibody, while each mouse in the control group received 100 μl (100 μg) of isotype control antibody via intraperitoneal injection twice a week for 6 weeks. Splenocytes (n = 4 in each group) were collected and stained for FACS analysis eight weeks after transplantation. The percentages and numbers of CD4⁺, CD69⁺CD4⁺T, CD8⁺, and CD69⁺CD8⁺T cells in H2Kb⁺ cells from the spleens of recipients are shown (A). Lymphocytes were isolated from the spleen and treated with PMA, brefeldin A, and ionomycin for 4-6 h. The percentage and number of TNF-α-(B) and IL-4-(C) positive T cells in the spleens of recipients are shown. The percentage and number of Treg (CD4⁺CD25⁺Foxp3⁺) cells in the spleens of recipients are shown (D). The percentage and number of CD4⁺CD8⁺T cells and the CD4⁺/CD4⁺CD8⁺ and CD8⁺/CD4⁺CD8⁺ ratios in the thymuses of recipients are shown (E). The percentage and number of Tfh (CD4⁺CXCR5⁺PD-1⁺) cells in the spleens of recipients are shown (F). The percentage and number of GC B (B220⁺GL-7⁺Fas⁺) cells in the spleens of recipients are shown (G). The data are representative of at least three independent experiments. Values are presented as mean ± SD. *P < 0.05; **P < 0.01