The control of hyperacute rejection by genetic engineering of the donor species.
Activation of endogenous complement is inhibited both in the soluble phase and at the membrane surface by a group of structurally similar proteins. A possible solution to hyperacute rejection is to produce donor animals transgenic for human complement regulators. Mouse cells expressing the human complement regulatory proteins decay accelerating factor (DAF) or membrane cofactor protein (MCP) were produced both by hybridoma technology and by transfection with the appropriate cDNAs. The expression of either or both of these products protected the mouse cell from lysis by human (though not rabbit) complement in the presence of naturally occurring human anti-mouse antibody. This effect could be abrogated by the addition of monoclonal antibody against DAF or MCP. Hyperacute rejection of discordant organ xenografts is mediated by human complement. A 6.5 kilobase minigene for DAF has been microinjected into porcine fertilised ova. Forty-five pigs transgenic for human DAF have been produced. Of these, 65% transcribe message. The amount of message produced varied substantially from animal to animal and was independent of copy number integrated. Expression of human DAF on the porcine lymphocyte surface could be detected and this was able to downregulate human complement activation. Amounts of protein expressed on different tissues varied both from pig to pig and within animals from tissue to tissue. The pigs grow and develop normally with no evidence of ill effects due to possession of the transgene.
White
D J
DJ
Department of Surgery, School of Clinical Medicine, University of Cambridge, UK.
Cozzi
E
E
Langford
G
G
Oglesby
T
T
Wang
M W
MW
Wright
L
L
Wallwork
J
J
D000818:Animals; D030801:Animals, Genetically Modified; D000911:Antibodies, Monoclonal / Q000276:immunology; D015703:Antigens, CD / Q000096:biosynthesis / Q000502:physiology*; D001483:Base Sequence; D001798:Blood Proteins / Q000096:biosynthesis; D018958:CD55 Antigens / Q000096:biosynthesis / Q000502:physiology*; D003169:Complement Inactivator Proteins; D005818:Genetic Engineering; D006084:Graft Rejection / Q000276:immunology / Q000517:prevention & control*; D006801:Humans; D006822:Hybrid Cells / Q000276:immunology; D008214:Lymphocytes / Q000276:immunology; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000502:physiology*; D051379:Mice; D008969:Molecular Sequence Data; D011817:Rabbits; D013552:Swine / Q000235:genetics; D014162:Transfection
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D001798:Blood Proteins; C497857:CD46 protein, human; D018958:CD55 Antigens; D003169:Complement Inactivator Proteins; C497859:Mcp protein, mouse; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins