Significant downregulation of the major swine xenoantigen by N-acetylglucosaminyltransferase III gene transfection.
Introduction of the beta-D-mannoside beta-1,4-N-acetylglucosaminyltransferase III (GnT-III) gene into swine endothelial cells (SEC) reduced their susceptibility to normal human serum (NHS) in complement-mediated cell lysis and also suppressed the antigenicity to human natural antibodies as evidenced by flow cytometric analysis, as well as Griffonia simplicifolia 1 isolectin (1B4 lectin) binding to the Gal alpha1-3 Gal beta 1-4 GlcNAc-R (the alpha-galactosyl epitope). Western blot analysis indicated that proteins smaller than 66 kDa had diminished reactivity to NHS and 1B4 lectin. GnT-III, a key enzyme involved in branch formation of N-linked sugars, was found to downregulate the expression of xenoantigen, suggesting that this approach may be of value in clinical xenotransplantation in the future.
Tanemura
M
M
Department of Biochemistry, Biomedical Research Center, Osaka University Medical School, Suita, Japan.
Miyagawa
S
S
Ihara
Y
Y
Matsuda
H
H
Shirakura
R
R
Taniguchi
N
N
D000818:Animals; D000906:Antibodies / Q000276:immunology; D015478:Antigens, Heterophile / Q000276:immunology / Q000378:metabolism*; D015153:Blotting, Western; D016923:Cell Death; D002460:Cell Line; D003165:Complement System Proteins / Q000276:immunology; D015536:Down-Regulation; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D005434:Flow Cytometry; D005647:Fucosyltransferases / Q000378:metabolism; D005690:Galactose / Q000276:immunology / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D037102:Lectins / Q000378:metabolism; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism*; D013552:Swine; D014162:Transfection; D000097631:Galactoside 2-alpha-L-fucosyltransferase
D000906:Antibodies; D015478:Antigens, Heterophile; D003165:Complement System Proteins; D000939:Epitopes; D005647:Fucosyltransferases; D005690:Galactose; D007770:L-Lactate Dehydrogenase; D037102:Lectins; D017351:N-Acetylglucosaminyltransferases; D000097631:Galactoside 2-alpha-L-fucosyltransferase; C054599:beta-1,4-mannosyl-glycoprotein beta-1,4-N-acetylglucosaminyltransferase