Abstract
We recently discovered that ABO incompatibility, which evokes a rapid humoral immune response in adult heart transplantation, is not a barrier in infant heart transplantation, and that infant recipients of ABO-incompatible hearts develop specific B-cell tolerance to donor A/B antigens. An animal model of ABO-incompatible heart transplantation would allow detailed investigation of the mechanism(s) of acceptance of ABO-incompatible grafts, using experimental methods that would not be possible in humans. To determine the feasibility of such a model, the human α-1,2-fucosyltransferase (H-transferase; for H antigen expression) gene was cloned into a lentiviral vector, and the human α-1,3-N-acetylgalactosaminyltransferase (A-transferase; for A antigen expression) gene was cloned into a bicistronic lentiviral vector also containing the green fluorescent protein (GFP) gene; these replication-deficient vectors were denoted H-trs and A-GFP, respectively. Synthesis of the human histo-blood group A antigen in humans is dependent on expression of these two glycosyltransferases. HeLa cells, a human cell line known to be of blood group O origin, expressed cell surface A antigen as measured by cellular ELISA when transfected with A-GFP alone, and the level of A antigen expression was enhanced by transfection with H-trs in addition to A-GFP. Cell surface H antigen expression was observed on both mouse fibroblast and mouse endothelial cells only when infected with H-trs lentiviral particles. Expression of A antigen was dependent on infection with both H-trs and A-GFP lentiviral particles, approaching levels on human group A cells. These collective results indicate that expression of human histo-blood group A antigen at the cell surface can be induced in mouse cells by infection with H-trs and A-GFP, and that such A antigen expression is dependent on H-trs expression.
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