Onor hearts transplanted into Fenofibrate-treated (100 mg/kg/d) recipients (median survival: 25 days; n = 6) compared to non-treated recipients (NT, median survival: 9.5 days; n = 6) (Wilcoxon log-rank test, p = 0.0007) (PD168393 biological activity Figure 4) and led to allograft beating beyond day 30 (study endpoint) in two animals. Mice tolerated the Fenofibrate dose well; no change in total body weight during the treatment was observed. AlloIndolactam V web Grafts in the NT control group were rejected within an average of 12 1655472 days after transplantation. Comparison to standard immunosuppression (Cyclosporine, Cys, 20 mg/kg/d i.p.) revealed equivalent efficacy of Fenofibrate for extending graft survival (median survival Cys 30 days) (Figure 4).Significantly improved graft function with Fenofibrate. Seven days daily treatment with Fenofibrate-using the same acute rejection model used for graft survival, resulted in significantly higher average BS’ (mean BS = 3.5) compared to NT (mean BS = 2, p,0.001) (Figure 5A). The chosen dosage of 100 mg/kg/d Fenofibrate was well tolerated by the mice as there was no change in body weight, liver function and serum Creatinine (Materials and Methods S1).Reduction in cellular infiltrates and myocyte damage with Fenofibrate. Graft tissue from POD7 was assessed bya single blinded pathologist by microscopic evaluation of H E staining applying ISHLT criteria [32,33]. Grafts from Fenofibrate treated animals revealed decreased cellular infiltrates either uni- or multifocal but never diffuse, and with- or without myocyte damage compared to grafts from untreated animals which always showed diffuse multifocal cellular infiltrates and were always associated with myocyte damage (Figure 5B).Significantly reduced numbers of graft infiltrating innate and adaptive immune cells with Fenofibrate. FACS analysisof graft infiltrating cells not only confirmed histological evaluation but further quantified the anti-inflammatory effect of Fenofibrate. Assessed were total numbers of graft infiltrating total leukocytes (CD45+), and of differential innate Dendritic- (DC,CD11c), Natural Killer (ND, NK1.1), Macrophages (F4/80), Neutrophils (Gr1+)) and adaptive immune Cytotoxic-(CD8), Helper T-(CD4), and B-(CD220)) cells in Fenofibrate and no-treated grafts at POD7 (Figure 5C). Numbers of cells were corrected for total graft weight. Fenofibrate significantly decreased the number of CD45+ graftDrug Repositioning Fenofibrate for TransplantationFigure 2. IL17 pathway- and IL17+ Th- gene sets in human AR: Segregation of AR and STA. Genes from the IL17 pathway 1317923 gene-set (2a) and from the Th17 gene-set (2b) were used for hierarchical clustering of post-transplant biopsies and resulted in clear separation of AR and STA after mean centering arrays and genes (Euclidean Distance, Cosine Dissimilarity). Unsupervised principal component analysis (PCA) of the same samples using the same genes confirmed the separation of AR and STA (2c, d). A separation of the AR group into C4d+ and C4d- cases was not seen. doi:10.1371/journal.pone.0056657.ginfiltrating total leukocytes (p,0.001); more specifically, there was a significant reduction in antigen presenting DC (p,0.01), in CD4+ T-helper and CD8+ cytotoxic T-cells, as well as in infiltrating neutrophils (p,0.01) and NK cells. As IL17 is known to promote neutrophil infiltration, the observed reduction of neutrophils in the graft of Fenofibrate treated mice potentially reflects the IL17 pathway inhibition by Fenofibrate. Infiltrating macrophages an.Onor hearts transplanted into Fenofibrate-treated (100 mg/kg/d) recipients (median survival: 25 days; n = 6) compared to non-treated recipients (NT, median survival: 9.5 days; n = 6) (Wilcoxon log-rank test, p = 0.0007) (Figure 4) and led to allograft beating beyond day 30 (study endpoint) in two animals. Mice tolerated the Fenofibrate dose well; no change in total body weight during the treatment was observed. Allografts in the NT control group were rejected within an average of 12 1655472 days after transplantation. Comparison to standard immunosuppression (Cyclosporine, Cys, 20 mg/kg/d i.p.) revealed equivalent efficacy of Fenofibrate for extending graft survival (median survival Cys 30 days) (Figure 4).Significantly improved graft function with Fenofibrate. Seven days daily treatment with Fenofibrate-using the same acute rejection model used for graft survival, resulted in significantly higher average BS’ (mean BS = 3.5) compared to NT (mean BS = 2, p,0.001) (Figure 5A). The chosen dosage of 100 mg/kg/d Fenofibrate was well tolerated by the mice as there was no change in body weight, liver function and serum Creatinine (Materials and Methods S1).Reduction in cellular infiltrates and myocyte damage with Fenofibrate. Graft tissue from POD7 was assessed bya single blinded pathologist by microscopic evaluation of H E staining applying ISHLT criteria [32,33]. Grafts from Fenofibrate treated animals revealed decreased cellular infiltrates either uni- or multifocal but never diffuse, and with- or without myocyte damage compared to grafts from untreated animals which always showed diffuse multifocal cellular infiltrates and were always associated with myocyte damage (Figure 5B).Significantly reduced numbers of graft infiltrating innate and adaptive immune cells with Fenofibrate. FACS analysisof graft infiltrating cells not only confirmed histological evaluation but further quantified the anti-inflammatory effect of Fenofibrate. Assessed were total numbers of graft infiltrating total leukocytes (CD45+), and of differential innate Dendritic- (DC,CD11c), Natural Killer (ND, NK1.1), Macrophages (F4/80), Neutrophils (Gr1+)) and adaptive immune Cytotoxic-(CD8), Helper T-(CD4), and B-(CD220)) cells in Fenofibrate and no-treated grafts at POD7 (Figure 5C). Numbers of cells were corrected for total graft weight. Fenofibrate significantly decreased the number of CD45+ graftDrug Repositioning Fenofibrate for TransplantationFigure 2. IL17 pathway- and IL17+ Th- gene sets in human AR: Segregation of AR and STA. Genes from the IL17 pathway 1317923 gene-set (2a) and from the Th17 gene-set (2b) were used for hierarchical clustering of post-transplant biopsies and resulted in clear separation of AR and STA after mean centering arrays and genes (Euclidean Distance, Cosine Dissimilarity). Unsupervised principal component analysis (PCA) of the same samples using the same genes confirmed the separation of AR and STA (2c, d). A separation of the AR group into C4d+ and C4d- cases was not seen. doi:10.1371/journal.pone.0056657.ginfiltrating total leukocytes (p,0.001); more specifically, there was a significant reduction in antigen presenting DC (p,0.01), in CD4+ T-helper and CD8+ cytotoxic T-cells, as well as in infiltrating neutrophils (p,0.01) and NK cells. As IL17 is known to promote neutrophil infiltration, the observed reduction of neutrophils in the graft of Fenofibrate treated mice potentially reflects the IL17 pathway inhibition by Fenofibrate. Infiltrating macrophages an.
