Classical pathway complement destruction is not responsible for the loss of human erythrocytes during porcine liver perfusion.
BACKGROUND
Porcine livers perfused with human blood destroy 85% of human erythrocytes (red blood cells [RBC]) during prolonged extracorporeal perfusion, raising the possibility of a complement-mediated graft-versus-host effect.


METHODS
Isolated porcine livers were perfused with fresh human blood. Plasma samples were analyzed for complement production by reverse CH50 analysis and porcine immunoglobulin class and specificity by enzyme-linked immunosorbent assay (ELISA) and flow cytometry. Anti-CD59 and anti-decay accelerating factor (DAF) monoclonal antibody were used to investigate whether human complement regulatory proteins inhibit porcine complement.


RESULTS
After 64 hr of perfusion of porcine livers with human blood, mean complement activity in the perfusate was 95% of the starting value and increasing, whereas perfusion in the absence of a liver showed a falling complement activity of 28.7%. ELISA demonstrated porcine immunoglobulin (Ig) G and IgM in the xenoperfused human plasma. Whereas in a previous study flow cytometry demonstrated porcine antibodies specific for antigens on human T lymphocytes, in this study, anti-human RBC antibodies were not found. Xenoperfused human plasma did not lyse fresh human RBC. Human complement was consistently more efficient at lysing porcine RBC than was porcine complement at lysing human RBC, and human plasma inhibited the ability of porcine plasma to lyse human RBC, raising the possibility of cross-species complement regulation. Complement regulatory proteins on human RBC were blocked using mouse monoclonal anti-human CD59 and DAF. Blocking CD59, but not DAF, augmented lysis of human RBC by porcine complement.


CONCLUSIONS
Human CD59 inhibits porcine complement. The production of porcine complement from xenoperfused porcine livers is unlikely to result in clinically significant injury mediated through the classical pathway of complement activation.
Rees
Michael A
MA
Department of Surgery, University of Cambridge, Addenbrooke's Hospital, Cambridge, Cambridgeshire, United Kingdom. mrees@mco.edu
Butler
Andrew J
AJ
Negus
Margaret C
MC
Davies
Hugh F S
HF
Friend
Peter J
PJ
D000818:Animals; D030801:Animals, Genetically Modified; D015478:Antigens, Heterophile / Q000276:immunology; D018958:CD55 Antigens / Q000276:immunology; D018957:CD59 Antigens / Q000276:immunology; D003171:Complement Pathway, Classical / Q000276:immunology*; D004912:Erythrocytes / Q000276:immunology*; D006086:Graft vs Host Disease / Q000276:immunology*; D006461:Hemolysis / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D008099:Liver / Q000276:immunology*; D010477:Perfusion; D013552:Swine
D015478:Antigens, Heterophile; D018958:CD55 Antigens; D018957:CD59 Antigens; D007074:Immunoglobulin G