Gene therapy for liver transplantation using adenoviral vectors: CD40-CD154 blockade by gene transfer of CD40Ig protects rat livers from cold ischemia and reperfusion injury.
Liver injury induced by ischemia/reperfusion (I/R) is the prime factor in delayed or loss graft function following transplantation. CD4+ T lymphocytes are key cellular mediators of antigen-independent inflammatory response triggered by I/R. We attempted to modulate rat liver I/R injury by targeted gene therapy with CD40Ig, which blocks the CD40-CD154 costimulation pathway. One hundred percent of Ad-CD40Ig-pretreated orthotopic liver transplants (OLTs) subjected to 24 h of cold (4 degrees C) ischemia survived > 14 days (vs 50% in untreated/Ad-beta-gal groups). Ad-CD40Ig treatment decreased sGOT levels and depressed neutrophil infiltration, compared with controls. These functional data correlated with histological Suzuki's grading of hepatic injury, which in untreated/Ad-beta-gal groups showed severe necrosis (> 60%) and moderate to severe sinusoidal congestion; the Ad-CD40Ig-pretreated group revealed minimal sinusoidal congestion/necrosis. Unlike in controls, OLT expression of mRNA coding for IL-2/IFN-gamma remained depressed, whereas that of IL-4/IL-13 reciprocally increased in the Ad-CD40Ig group. Ad-CD40Ig reduced frequency of TUNEL+ cells and pro-apoptotic Caspase-3, but enhanced antioxidant HO-1 and anti-apoptotic Bcl-2/Bcl-xl expression. Thus, prolonged blockade of CD40-CD154 by CD40Ig exerts potent cytoprotection against hepatic I/R injury. These results provide the rationale for a novel gene therapy approach to maximize the organ donor pool through the safer use of liver transplants exposed to prolonged cold ischemia.
Ke
Bibo
B
Dumont--UCLA Transplant Center and David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA 90095, USA.
Shen
Xiu-Da
XD
Gao
Feng
F
Busuttil
Ronald W
RW
Löwenstein
Pedro R
PR
Castro
Maria G
MG
Kupiec-Weglinski
Jerzy W
JW
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D017209:Apoptosis; D019013:CD40 Antigens / Q000235:genetics*; D023201:CD40 Ligand / Q000502:physiology; D003080:Cold Temperature; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D007136:Immunoglobulins / Q000235:genetics; D008107:Liver Diseases / Q000209:etiology / Q000473:pathology / Q000517:prevention & control*; D016031:Liver Transplantation / Q000379:methods* / Q000473:pathology; D008297:Male; D020556:Neutrophil Infiltration; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000209:etiology / Q000473:pathology / Q000517:prevention & control*
D019013:CD40 Antigens; D016207:Cytokines; D007136:Immunoglobulins; D023201:CD40 Ligand