Anti-VEGFR-3 treatment ameliorates lethal GVHD. Allo-HSCT recipients received intraperitoneal injections of 1 mg anti-VEGFR-3 antibody (mF4-31c1) or control antibody every second day from day 0 to day +16. LP/J or 129/SV → C57BL/6, conditioning with Bu/Cy; allo-HSCT with 1.5 × 107 BM cells, 2 × 106 T cells. (A) Survival curve of control antibody and mF4-31c1 treated allo-HSCT recipients using the 129/SV → C57BL/6 model (n = 5 per group), data from one representative experiment are shown, analysis with the log-rank test. (B) GVHD scores of control antibody and mF4-31c1–treated allo-HSCT recipients on day +14 using the 129/SV → C57BL/6 model. Data from one representative experiment are shown (n = 5 per group). (C) Survival curve in the LP/J → C57BL/6 model. Combined data from three independent experiments are presented (n = 28 per group). Analysis was done with the log-rank test. (D) GVHD scores of control antibody versus mF4-31c1–treated allo-HSCT recipients in the LP/J → C57BL/6 model on day +18 after HSCT. Combined data from three independent experiments are presented (n = 28 per group), analysis with the log-rank test. (E) Representative images of histopathology in the colon and liver. Organs were taken on day +11 after HSCT; histological staining was performed with hematoxylin and eosin (n = 4 per group). (F) Histopathological scores of colon and liver sections from control antibody and mF4-31c1–treated allo-HSCT recipients on day +11 after HSCT; scoring was done according to Lerner criteria (n = 4 per group). (G) Representative images of CD3+ cell infiltration in the liver. Organs were taken on day +11, liver sections were stained with CD3 antibody (green) and counterstained with DAPI. (H) Quantification of CD3 positive area in the liver of control antibody or mF4-31c1–treated mice (n = 4 per group). Error bars indicate mean ± SEM; significance was tested with an unpaired Student t test. BMT, bone marrow transplantation.
