Expression of the human alpha1,2-fucosyltransferase in transgenic pigs modifies the cell surface carbohydrate phenotype and confers resistance to human serum-mediated cytolysis.
Hyperacute rejection (HAR) is the first critical immunological hurdle that must be addressed in order to develop xenogeneic organs for human transplantation. In the area of cell-based xenotransplant therapies, natural antibodies (XNA) and complement have also been considered barriers to successful engraftment. Transgenic expression of human complement inhibitors in donor cells and organs has significantly prolonged the survival of xenografts. However, expression of complement inhibitors without eliminating xenogeneic natural antibody (XNA) reactivity may provide insufficient protection for clinical application. An approach designed to prevent XNA reactivity during HAR is the expression of human alpha1, 2-fucosyltransferase (H-transferase, HT). H-transferase expression modifies the cell surface carbohydrate phenotype of the xenogeneic cell, resulting in the expression of the universal donor O antigen and a concomitant reduction in the expression of the antigenic Galalpha1,3-Gal epitope. We have engineered various transgenic pig lines that express HT in different cells and tissues, including the vascular endothelium. We demonstrate that in two different HT transgenic lines containing two different HT promoter constructs, expression can be differentially regulated in a constitutive and cytokine-inducible manner. The transgenic expression of HT results in a significant reduction in the expression of the Galalpha1,3-Gal epitope, reduced XNA reactivity, and an increased resistance to human serum-mediated cytolysis. Transgenic pigs that express H-transferase promise to become key components for the development of xenogeneic cells and organs for human transplantation.
Costa
C
C
Department of Molecular Sciences, Alexion Pharmaceuticals Inc, New Haven, Connecticut 06511, USA.
Zhao
L
L
Burton
W V
WV
Bondioli
K R
KR
Williams
B L
BL
Hoagland
T A
TA
Ditullio
P A
PA
Ebert
K M
KM
Fodor
W L
WL
D000017:ABO Blood-Group System / Q000276:immunology; D000818:Animals; D030801:Animals, Genetically Modified; D001011:Aorta / Q000276:immunology; D002462:Cell Membrane / Q000276:immunology; D004730:Endothelium, Vascular / Q000276:immunology; D005347:Fibroblasts / Q000276:immunology; D005647:Fucosyltransferases / Q000096:biosynthesis* / Q000235:genetics; D006084:Graft Rejection / Q000097:blood*; D006801:Humans; D008562:Membrane Glycoproteins / Q000276:immunology; D010641:Phenotype; D013552:Swine / Q000235:genetics* / Q000276:immunology*; D014183:Transplantation, Heterologous / Q000276:immunology*; D000097631:Galactoside 2-alpha-L-fucosyltransferase
D000017:ABO Blood-Group System; D005647:Fucosyltransferases; D008562:Membrane Glycoproteins; D000097631:Galactoside 2-alpha-L-fucosyltransferase