The Effect of Leflunomide Analogue FK778 on Development of Chronic Rat Renal Allograft Rejection and Transforming Growth Factor-BETA Expression

Abstract

Background Chronic allograft nephropathy (CAN) remains the primary reason for late allograft loss in kidney transplantation. Transforming growth factor beta (TGF-β) is a major mitogen mediating mesenchymal cell proliferation and epithelial to mesenchymal cell transition in CAN. FK778, an analogue of an active metabolite of leflunomide, is a promising immunosuppressive drug that inhibits de novo pyrimidine biosynthesis. Herein we investigated the effect of FK778 on development of chronic rejection and TGF-β expression in combination with calcineurin inhibitors cyclosporine (CsA) and tacrolimus (Tac). Methods Kidney transplantations were performed from DA to WF rats and syngeneic control transplantations between DA rats. Allografts were immunosupressed alone with CsA (1.5 mg/kg/d subcutaneously) or Tac (1.5 mg/kg/d orally) or with combinations of FK778 (10 mg/kg/d orally) and CsA or Tac. No immunosuppression was given to syngeneic grafts. Grafts were harvested 90 days after transplantation for histology and immunohistochemistry (TGF-β, TGF-βR1). The chronic changes in allografts were scored according to the Chronic Allograft Damage Index (CADI). Results No histological signs of chronic rejection were seen in syngeneic grafts. According to CADI, moderate chronic changes were seen in grafts treated only with CsA or Tac. In both groups the changes typically associated with CAN were significantly ameliorated with FK778. CsA-treated grafts showed intense posttransplant expression of TGF-β and TGF-βR1 after 90 days. In grafts treated with Tac monotherapy this expression was substantially lower. FK778 markedly reduced the expression of TGF-β and TGF-βR1 when combined with calcineurin inhibitors and lesser expression was demonstrated with the combination of FK778 and Tac. Conclusions Our results demonstrated that FK778 is a potent immunosuppressive drug having synergistic effects with calcineurin inhibitors. When combined with CsA or Tac, it decreased posttransplant TGF-β ligand and receptor expression. Our data also showed that FK778 prevented chronic changes typically associated with CAN. Taken together our results suggested that FK778 could be a promising therapy for CAN in clinical kidney transplantation.