The Effect of Recombinant Human Alpha-1,2-Fucosyltransferase and Alpha-Galactosidase A on the Reduction of Alpha-Gal Expression in the Liver of Transgenic Pigs

Genetically modified pigs lacking Galα1 → 3Gal and other immunogenic carbohydrates are considered as the most promising, alternative source of various tissues and organs for human transplantation. Here, we tested the hypothesis that combining the expression of human α1,2-fucosyltransferase (hFUT2) and α-galactosidase A (hGLA) genes would allow for the removal of this specific carbohydrate in porcine transgenic livers. We investigated the expression profile of human α1,2-fucosyltransferase and α-galactosidase A proteins and the amount of Galα1 → 3Gal antigen in the liver of single transgenic hFUT2 (n=5), hGLA (n=5), and double transgenic hFUT2 × hGLA (n=5) pigs. Both human proteins, α1,2-fucosyltransferase and α-galactosidase A, were abundantly expressed in the liver tissue in respective transgenic lines as was revealed by confocal microscopy and Western blotting. The level of Galα1 → 3Gal epitope evaluated by lectin histochemistry and lectin blotting was significantly lower (p