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- Title
The alpha1,3GalT knockout/alpha1,2FucT transgenic pig does not appear to have an advantage over the alpha1,3GalT knockout pig with respect to glycolipid reactivity with human serum antibodies.
- Authors
Diswall, Mette; Benktander, John; Ångström, Jonas; Teneberg, Susann; Breimer, Michael E.
- Abstract
Background The human H-transferase (α2FucT) was introduced in Gal-negative pigs to produce pig organs not only free from Gal-antigens, but also in which the uncapped N-acetyllactosamine precursor had been transformed into non-xenogenic blood group H type 2 compounds. This work is the first descriptive analysis of glycolipids from the GalT- KO/FucT- TG pig. The aim was to investigate the cell membrane antigens in GalT- KO/FucT- TG tissues to explore its efficacy as an organ donor. Also, detailed knowledge on the correlation between the cellular glycosyltransferase configuration and the resulting carbohydrate phenotype expression is valuable from a basic glycobiological perspective. Methods Neutral and acidic glycolipids from GalT- KO/FucT- TG small intestine were compared with glycolipids from two wildtype and two GalT- KO pig intestines. Glycolipid reactivity was tested on thin layer chromatography plates using chemical reagents, antibodies, lectins, and human serum. Structural characterization of neutral glycolipids was performed by LC- ESI/ MS and proton NMR spectroscopy. Results Characterization of the glycolipid expression in GalT- KO/FucT- TG intestine showed absence of Gal antigens and decreased/unchanged levels of the N-acetyllactosamine precursor and the blood group H type 2 expression, when compared with the wildtype. The reactivity of human serum antibodies to GalT- KO/FucT- TG derived glycolipids was similar or slightly elevated when compared with GalT- KO glycolipids. Results from LC- ESI/ MS and proton NMR spectroscopy revealed no established neutral xenogenic antigens in the GalT- KO/FucT- TG pig, and could thus not explain the immunologic reactivity to human serum antibodies. The antibody binding to acidic glycolipids is most likely to be explained by the abundance of N-glycolylneuraminic acid epitopes in pig tissues. Six neutral complex biantennary glycolipids with blood group H type 1, 2, Lewisx and Lewisy determinants were found, of which three were identified in this work for the first time. One of these was a nonaglycosylceramide with blood group H type 2 and lactosyl determinants linked to a lactotetraosyl core, and the other two were decaglycosylceramides with blood group H type 1 and H type 2 determinants linked to a neolactotetraosyl core, and Lewisx and blood group H type 1 determinants on a lactotetraosyl core, respectively. Conclusions Lipid-linked carbohydrate antigens in the GalT- KO/FucT- TG pig intestine showed no or minor qualitative difference when compared with GalT- KO pigs. The GalT- KO/FucT- TG pig did not appear to have an advantage over the GalT- KO pig with respect to reactivity with human antibodies from a xenotransplantation perspective.
- Subjects
TRANSFERASES; GLYCOLIPIDS; GLYCOCONJUGATES; LIPIDS; IMMUNOGLOBULINS
- Publication
Xenotransplantation, 2014, Vol 21, Issue 1, p57
- ISSN
0908-665X
- Publication type
Article
- DOI
10.1111/xen.12071