STAT-1 decoy oligodeoxynucleotide inhibition of acute rejection in mouse heart transplants.
During acute rejection of cardiac transplants endothelial cell-leukocyte interaction fueled by co-stimulatory molecules like CD40/CD154 may ultimately lead to graft loss. One key player in up-regulating the expression of such pro-inflammatory gene products is the interferon-gamma-dependent transcription factor STAT-1. Hence down-regulating interferon-gamma-stimulated pro-inflammatory gene expression in the graft endothelial cells by employing a decoy oligodeoxynucleotide (dODN) neutralising STAT-1 may protect the graft. To verify this hypothesis, heterotopic mouse heart transplantation was performed in the allogeneic B10.A(2R) to C57BL/6 and syngeneic C57BL/6 to C57BL/6 strain combination without immunosuppression. Graft vessels were pre-treated with STAT-1 dODN, mutant control ODN (10 muM each) or vehicle (Ringer solution). Cellular rejection (vascular and interstitial component) was graded histologically and CD40, ICAM-1, VCAM-1, MCP-1, E-selectin and RANTES expression in the graft monitored by real time PCR 24 h and 9 days post-transplantation. Nine days after transplantation both rejection scores were significantly diminished by 85 and 70%, respectively, in STAT-1 dODN-treated allografts as compared to mutant control ODN-treated allografts. According to immunohistochemistry analysis, this was accompanied by a reduced infiltration of monocyte/macrophages and T cells into the graft myocardium. In addition, pro-inflammatory gene expression was strongly impaired by more than 80% in STAT-1 dODN-treated allografts 24 h post-transplantation but not in mutant control ODN or vehicle-treated allografts. This inhibitory effect on pro-inflammatory gene expression was no longer detectable 9 days post-transplantation. Single periprocedural treatment with a STAT-1 dODN thus effectively reduces cellular rejection in mouse heart allografts. This effect is associated both with an early decline in pro-inflammatory gene expression and a later drop in mononuclear cell infiltration.
Stojanovic
Tomislav
T
Department of Heart, Thoracic and Vascular Surgery, University of Göttingen, Robert-Koch-Str 40, 37073, Göttingen, Germany.
Wagner
Andreas H
AH
Wang
Shijun
S
Kiss
Eva
E
Rockstroh
Nicolas
N
Bedke
Jens
J
Gröne
Hermann-Josef
HJ
Hecker
Markus
M
D000818:Animals; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics; D015536:Down-Regulation; D015870:Gene Expression / Q000187:drug effects* / Q000235:genetics; D006084:Graft Rejection / Q000235:genetics / Q000473:pathology / Q000517:prevention & control*; D016027:Heart Transplantation / Q000473:pathology; D007150:Immunohistochemistry; D007249:Inflammation / Q000235:genetics / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D009841:Oligonucleotides / Q000494:pharmacology* / Q000627:therapeutic use; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050794:STAT1 Transcription Factor / Q000378:metabolism*
D015815:Cell Adhesion Molecules; D009841:Oligonucleotides; D050794:STAT1 Transcription Factor; C494080:Stat1 protein, mouse