Prevention of complement-mediated activation of xenogeneic endothelial cells in an in vitro model of xenograft hyperacute rejection by C1 inhibitor.
The complement system plays a major role in hyperacute rejection of discordant xenografts. In an immediately vascularized xenograft of a porcine organ to a primate, natural antibodies bind to the vascular endothelium of the graft, triggering activation of complement via the classical pathway. One consequence of antibody binding and complement activation is the activation of endothelial cells leading to the loss from the cells of heparan sulfate. We explored to what extent the classical pathway regulatory protein C1 inhibitor (C1 inh) would inhibit complement-mediated cytotoxicity and activation of endothelial cells. Cultured porcine aortic endothelial cells were used as a model for a xenogeneic organ and human serum as a source of xenoreactive natural antibody and complement. Addition of purified human C1 inh to human serum inhibited deposition of C4b and iC3b and cytotoxicity after the serum was reacted with the cultured cells. C1 inh prevented, in a dose-dependent manner, activation of the endothelial cells, as manifested by release of heparan sulfate. These observations demonstrate that C1 inh added in sufficient amounts to human serum can effectively inhibit C1 activation in an antigen-antibody system. These studies extend our previous findings consistent with the concept that complement activation occurs via the classical pathway in models of hyperacute rejection in which porcine vascular endothelial cells are in contact with human serum containing xenogeneic natural antibodies against the endothelial cells. Thus, our results suggest a potential clinical use of C1 inh in conjunction with other therapies to prevent hyperacute rejection in xenogeneic combinations mediated by complement activation via the classical pathway.
Dalmasso
A P
AP
Laboratory Service, Veterans Affairs Medical Center, Minneapolis, MN 55417.
Platt
J L
JL
D000818:Animals; D002478:Cells, Cultured; D003167:Complement Activation; D003174:Complement C1 Inactivator Proteins / Q000494:pharmacology*; D003165:Complement System Proteins / Q000502:physiology*; D003602:Cytotoxicity, Immunologic; D004730:Endothelium, Vascular / Q000276:immunology*; D006084:Graft Rejection; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D013552:Swine; D014183:Transplantation, Heterologous / Q000276:immunology*
D003174:Complement C1 Inactivator Proteins; D003165:Complement System Proteins; D006497:Heparan Sulfate