The effect of leflunomide analogue FK778 on development of chronic rat renal allograft rejection and transforming growth factor-BETA expression.
BACKGROUND
Chronic allograft nephropathy (CAN) remains the primary reason for late allograft loss in kidney transplantation. Transforming growth factor beta (TGF-beta) 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-beta 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-beta, TGF-betaR1). 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-beta and TGF-betaR1 after 90 days. In grafts treated with Tac monotherapy this expression was substantially lower. FK778 markedly reduced the expression of TGF-beta and TGF-betaR1 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-beta 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.
Rintala
J M
JM
Transplantation Laboratory, University of Helsinki, and Helsinki University Central Hospital, Helsinki, Finland. jukka.m.rintala@helsinki.fi
Savikko
J
J
Rintala
S E
SE
von Willebrand
E
E
D000480:Alkynes / Q000627:therapeutic use*; D000818:Animals; D016572:Cyclosporine / Q000627:therapeutic use; D004359:Drug Therapy, Combination; D006084:Graft Rejection / Q000517:prevention & control*; D007166:Immunosuppressive Agents / Q000627:therapeutic use*; D007555:Isoxazoles / Q000627:therapeutic use*; D016030:Kidney Transplantation / Q000276:immunology*; D009570:Nitriles / Q000627:therapeutic use*; D051381:Rats; D011920:Rats, Inbred WF; D016559:Tacrolimus / Q000627:therapeutic use; D016212:Transforming Growth Factor beta / Q000235:genetics*; D014184:Transplantation, Homologous / Q000276:immunology*; D014185:Transplantation, Isogeneic
D000480:Alkynes; D007166:Immunosuppressive Agents; D007555:Isoxazoles; D009570:Nitriles; D016212:Transforming Growth Factor beta; C115284:2-cyano-3-hydroxy-N-(4-(trifluoromethyl)phenyl)-2-hepten-6-ynamide; D016572:Cyclosporine; D016559:Tacrolimus