Hyperacute rejection of a porcine organ by higher primates is initiated by the binding of xenoreactive natural antibodies of the recipient to blood vessels in the graft leading to complement activation. The majority of these antibodies recognize the carbohydrate structure Gal(al,3)Gal (gal epitope) present on cells of pigs. It is possible that the removal or lowering of the number of gal epitopes on the graft endothelium could prevent hyperacute rejection. The Gal(al,3)Gal structure is formed by the enzyme Gal81,4GlcNAc3-i-

Fetal-pig fibroblasts homozygous for the knockout of the alpha1-3 galactosyltransferase gene appear to express low but detectable levels of the gal antigen.

We describe isologous promoter replacement as an approach to permit high level expression of human hemoglobin in transgenic swine. We linked the human beta globin genomic coding region to the porcine beta globin promoter and used this fusion gene in an expression construct containing the human beta locus control region and the human alpha and epsilon genes to produce transgenic pigs. The highest level of expression was 24% human (32g/liter) and 30% human alpha/pig beta hybrid (40g/liter) hemoglobin in one transgenic pig. This pig was bred to a non-transgenic animal resulting in the transmission of high level human hemoglobin expression to 5 of 12 progeny.

These observations suggest that this model can be used to study the regulation of anti-Gal B cells and can establish a reliable source of functional anti-Gal B cells, which could be used to test the effectiveness of alpha-Gal-specific immunosuppressive reagents.

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