Effects of prolonged cold storage time in xenotransplantation.
BACKGROUND
Ischemia and reperfusion injury after prolonged ischemic time (IT) has a marked influence on hyperacute xenograft rejection (HXR). The aim of the study was to investigate the impact of different cold ischemic times on the HXR of ex vivo "working pig hearts" perfused with human blood. Xenoreactive natural antibodies (XNAb) as the trigger of HXR were reduced by immunoadsorption (IA) using an Ig-Therasorb column.


METHODS
Hearts of 24 Landrace pigs (13-31 kg) were harvested after cardioplegia with Celsior-solution and split into 4 groups. In Group C1 (n = 6) the short ischemic time (IT) lasted 48.9+/-10 minutes on the average prior to start of xenoperfusion, in Group I1 IT lasted 4 hours instead. Groups C2 and 12 experienced the same IT as C1 and I1, respectively, but underwent 2 cycles of IA in addition. IA removed immunoglobulins IgG, IgM and IgA from the perfusate. In the working heart mode hemodynamic parameters were measured in defined intervals. Blood samples were collected at the same time to determine myocardial enzymes, immunoglobulins, complement and anti-pig-antibodies. At the end of the study (cardiac arrest) tissue was sampled for histologic examination (light/electron microscopy (LM/EM) and immunohistochemistry).


RESULTS
Survival time of the control Group C1 was 125 minutes. IA resulted in an extension of perfusion time to 6.5 hours in C2. Four hours of IT (I1) prolonged the working time of hearts when compared with C1. IA had no additional impact (I2). Heart weight increased significantly in C1 without IA. Cardiac output and coronary flow in C1 were significantly lower when compared with the other 3 groups. IA improved cardiac output (CO) in C2 (vs C1, p < 0.001). Histologic signs of HXR (LM/EM) could be found in C1 in contrast to the other groups. Serologic parameters for myocardial damage were higher in groups with prolonged IT than in groups with short IT.


CONCLUSIONS
Prolonged ischemia and reperfusion injury showed controversial effects in this specific xenogeneic heart transplant model. In contrast to observations in allogeneic transplantation 4 hours of IT showed a beneficial behaviour of the xenografts. Reasons could be either a protective effect of the Celsior solution or changes of the endothelial cell surface (in terms of glycosylation or loss of alpha1-3Gal-epitopes). Tolerance of prolonged IT would allow transportation of xenografts over long distances.
Brenner
P
P
Department of Cardiac Surgery, Klinikum Grosshadern, Ludwig-Maximilians-University of Munich, Germany. Paolo.Brenner@hch.med.uni-muenchen.de
Hinz
M
M
Huber
H
H
Schmoeckel
M
M
Reichenspurner
H
H
Meiser
B
B
Hammer
C
C
Reichart
B
B
D000818:Animals; D000906:Antibodies / Q000032:analysis; D003080:Cold Temperature; D003165:Complement System Proteins / Q000032:analysis; D004187:Disaccharides / Q000494:pharmacology; D004573:Electrolytes / Q000494:pharmacology; D005971:Glutamates / Q000494:pharmacology; D005978:Glutathione / Q000494:pharmacology; D016027:Heart Transplantation / Q000379:methods*; D006439:Hemodynamics / Q000502:physiology; D006639:Histidine / Q000494:pharmacology; D006801:Humans; D007136:Immunoglobulins / Q000032:analysis; D007163:Immunosorbent Techniques; D008353:Mannitol / Q000494:pharmacology; D015428:Myocardial Reperfusion Injury / Q000503:physiopathology*; D009206:Myocardium / Q000276:immunology / Q000473:pathology; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions / Q000494:pharmacology; D013552:Swine; D013997:Time Factors; D014183:Transplantation, Heterologous
D000906:Antibodies; C088453:Celsior; D004187:Disaccharides; D004573:Electrolytes; D005971:Glutamates; D007136:Immunoglobulins; D019188:Organ Preservation Solutions; D008353:Mannitol; D006639:Histidine; D003165:Complement System Proteins; D005978:Glutathione