Monoclonal antibodies directed against human C5 and C8 block complement-mediated damage of xenogeneic cells and organs.
The hyperacute rejection (HAR) of xenotransplanted organs is initiated by the deposition of natural antibodies on donor endothelium followed by the activation of the recipient complement system, which rapidly destroys the graft. Studies of the role of activated complement in HAR have suggested that natural antibody as well as early (C3a, C3b) and late (C5a, C5b-9) activated complement components may contribute to cell activation and damage. Attenuation of HAR has been achieved by blockade of C3 activation with soluble CR1 or consumptive depletion of complement with cobra venom factor; however, similar studies using specific inhibitors of terminal complement components have not been described. To address the contribution of C5a and the membrane attack complex (C5b-9, MAC) to complement-mediated xenogeneic cell and organ damage, we utilized functionally blocking monoclonal antibodies directed against the human terminal complement components C5 and C8. Our data show that both anti-C5 and anti-C8 mAbs protect porcine aortic endothelial cells from membrane damage mediated by human C5b-9. Additionally, both the anti-C5 and anti-C8 mAbs blocked complement-mediated generation of membrane prothrombinase activity on porcine aortic endothelial cells challenged with human serum. To test the ability of these antibodies to attenuate antibody and complement-mediated damage of xenogeneic organs, an ex vivo model was developed wherein isolated rat hearts were perfused with human serum in the presence or absence of the anti-C5 and anti-C8 mAbs. Our data demonstrate that mAbs directed against human C5 and C8 prevented organ damage by human serum complement and suggest that these molecules may serve as potent inhibitors of HAR.
Department of Immunobiology, Alexion Pharmaceuticals, Inc., New Haven, Connecticut 06511, USA.
S A
SA
Rollins
L A
LA
Matis
J P
JP
Springhorn
E
E
Setter
D W
DW
Wolff
D000818:Animals; D000911:Antibodies, Monoclonal; D003167:Complement Activation; D003182:Complement C5 / Q000276:immunology*; D003185:Complement C8 / Q000276:immunology*; D015938:Complement Membrane Attack Complex / Q000276:immunology* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000276:immunology; D006084:Graft Rejection; D006801:Humans; D009206:Myocardium / Q000276:immunology; D010477:Perfusion; D051381:Rats; D013552:Swine; D013925:Thromboplastin / Q000378:metabolism
D000911:Antibodies, Monoclonal; D003182:Complement C5; D003185:Complement C8; D015938:Complement Membrane Attack Complex; D013925:Thromboplastin