The influence of antibody and complement removal with a Ig-Therasorb column in a xenogeneic working heart model.
OBJECTIVE
Organ transplantation is limited by the number of brain-dead human donors. Xenotransplantation could be an alternative to guarantee a constant supply of organs. A major problem of xenotransplantation are xenogeneic natural antibodies (XNAb) directed against species-specific antigens of a discordant donor species (e.g. pig). They trigger the hyperacute xenograft rejection (HXR). Re-usable immunoapheresis (LA)-columns Ig-Therasorb (Therasorb, Baxter) were used to adsorb these XNAb. The effect of immunoapheresis of the perfusing human blood was investigated in ex vivo working pig hearts.


METHODS
Hearts of 12 landrace pigs (body weight 14-31 kg) were explanted after inducing cardiac arrest with 4 degrees C Celsior solution. Human blood (500 ml, heparinized) was obtained from healthy volunteers. In group 1 (G1, n = 6), blood as perfusate remained untreated. In group 2 (G2, n = 6), native blood was separated by plasmapheresis into cellular components and plasma. The latter passed through the Ig-Therasorb column for removal of immunoglobulins (so-called immunoadsorption or immunoapheresis). After back-table preparation the hearts were mounted to the working heart model. After 20 min of reperfusion in Langendorff mode, the working heart mode was established. Blood samples were taken isochronously for measurement of: CK(-MB), LDH, ASAT, troponin, immunoglobulins, complement activity, anti-pig antibodies and others. After cessation of the heart, atrial and ventricular tissue samples were taken for histological examinations (light/electron microscopy and immunohistochemistry).


RESULTS
Two cycles of immunoapheresis reduced the levels of IgG by 84%, IgM by 83.3% and IgA by 76%. In G2, the antibody immunoadsorption of blood prolonged the duration of the working heart mode significantly to 335+/-37.5 min. In contrast, hearts of group 1 (control) failed after 125+/-31.3 min. Heart rate was significantly different between both groups (G1, 77.3+/-6.1 beats/min; G2, 86.5+/-5.5 beats/min). In G2 cardiac output was 118% and mean coronary flow was 154.6% higher than in G1. CK, LDH and ASAT showed no differences in the two groups. Heart weight increased significantly more in group 1 than in G2. Histological examination indicated specific signs of HXR in G1 after 1.5 h, whereas in G2 only slight unspecific damages were found after 6 h.


CONCLUSIONS
Antibody removal by means of immunoapheresis results in a significantly improved xenogeneic cardiac function. Immunoapheresis may, therefore, become an important adjunct in future pig-to-man clinical xenotransplantation.
Brenner
P
P
Department of Cardiac Surgery, Klinikum Grosshadern, Ludwig-Maximilians-University Munich, Germany.
Hinz
M
M
Huber
H
H
Schmoeckel
M
M
Reichenspurner
H
H
Meiser
B
B
Hammer
C
C
Reichart
B
B
D000818:Animals; D005112:Extracorporeal Circulation / Q000379:methods*; D016027:Heart Transplantation / Q000276:immunology* / Q000473:pathology*; D006439:Hemodynamics / Q000502:physiology; D006801:Humans; D007136:Immunoglobulins / Q000097:blood*; D007163:Immunosorbent Techniques; D007518:Isoantibodies / Q000097:blood; D008297:Male; D008928:Mitochondria / Q000648:ultrastructure; D008955:Models, Cardiovascular; D009206:Myocardium / Q000276:immunology* / Q000473:pathology*; D012016:Reference Values; D012508:Sarcolemma / Q000648:ultrastructure; D013045:Species Specificity; D013552:Swine; D014183:Transplantation, Heterologous
D007136:Immunoglobulins; D007518:Isoantibodies