Comparative analysis of three genetic modifications designed to inhibit human serum-mediated cytolysis.
Hyperacute rejection (HAR) remains a critical immunologic hurdle in the development of xenogeneic organs for human transplantation. Strategies that simultaneously eliminate both natural antibody reactivity and complement activation on the xenogeneic cell surface may be the best approach to achieve clinical application of xenogeneic vascularized organ transplantation. We have developed multiple lines of genetically manipulated mice to evaluate the combination of different genetic approaches aimed at inhibiting antibody and complement-mediated cell lysis. We utilized transgenic mice expressing the human complement inhibitor, CD59, the human 1,2-fucosyltransferase (H-transferase, HT) and the alpha1,3-galactosyltransferase (alpha1,3-GT) knock-out mouse line (Gal KO). Our data show that expression of hCD59 in combination with HT expression or the null phenotype of alpha1,3-GT are equally effective at preventing human serum-mediated cytolysis. Interestingly, the triple combination affords no additional protective effect. Therefore, coexpression of HT and a complement inhibitor is the most immediate strategy to genetically engineer transgenic pigs to be used as xenogeneic donors.
Costa
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Department of Molecular Sciences, Alexion Pharmaceuticals Inc., New Haven, CT 06511, USA.
Zhao
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Decesare
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Fodor
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D000818:Animals; D000910:Antibodies, Heterophile / Q000096:biosynthesis; D001483:Base Sequence; D018957:CD59 Antigens / Q000235:genetics; D003165:Complement System Proteins / Q000378:metabolism; D003602:Cytotoxicity, Immunologic / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D005647:Fucosyltransferases / Q000235:genetics; D005700:Galactosyltransferases / Q000235:genetics; D006084:Graft Rejection / Q000235:genetics*; D006801:Humans; D007113:Immunity, Innate; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D014183:Transplantation, Heterologous; D000097631:Galactoside 2-alpha-L-fucosyltransferase
D000910:Antibodies, Heterophile; D018957:CD59 Antigens; D003165:Complement System Proteins; D017931:DNA Primers; D005647:Fucosyltransferases; D005700:Galactosyltransferases; D000097631:Galactoside 2-alpha-L-fucosyltransferase; C047006:N-acetyllactosaminide alpha-1,3-galactosyltransferase