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Diswall M, Ångström J, Karlsson H, Phelps CJ, Ayares D, Teneberg S, Breimer ME. Structural characterization of α1,3‐galactosyltransferase knockout pig heart and kidney glycolipids and their reactivity with human and baboon antibodies. Xenotransplantation 2010; 17: 48–60. © 2010 John Wiley & Sons A/S. Abstract: Background: α1,3‐galactosyltranferase knockout (GalT‐KO) pigs have been established to avoid hyperacute rejection in GalT‐KO pig‐to‐human xenotransplantation. GalT‐KO pig heart and kidney glycolipids were studied focusing on elimination of Gal‐antigens and whether novel antigens would appear. Non‐human primates are used as pre‐clinical transplantation experimental models. Therefore, sera from baboons transplanted with GalT‐KO hearts were compared with human serum regarding reactivity with pig glycolipids. Methods: Neutral and acidic glycolipids were isolated from GalT‐KO and WT pig hearts and kidneys. Glycolipid immune reactivity was tested on TLC plates using human affinity‐purified anti‐Gal Ig, anti‐blood group monoclonal antibodies, lectins, and human serum as well as baboon serum collected before and after GalT‐KO pig heart transplantations. Selected glycolipid fractions, isolated by HPLC, were structurally characterized by mass spectrometry and proton NMR spectroscopy. Results: GalT‐KO heart and kidney lacked α3Gal‐terminated glycolipids completely. Levels of uncapped N‐acetyllactosamine precursor compounds, blood group H type 2 core chain compounds, the P1 antigen and the x2 antigen were increased. Human serum antibodies reacted with Gal‐antigens and N‐glycolylneuraminic acid (NeuGc) in WT organs of which only the NeuGc reactivity remained in the GalT‐KO tissues. A clear difference in reactivity between baboon and human antibodies with pig glycolipids was found. This was most pronounced for acidic, not yet identified, compounds in GalT‐KO organs which were less abundant or lacking in the corresponding WT tissues. Conclusions: GalT‐KO pig heart and kidney completely lacked Gal glycolipid antigens whilst glycolipids synthesized by competing pathways were increased. Baboon and human serum antibodies showed a different reactivity pattern to pig glycolipid antigens indicating that non‐human primates have limitations as a human pre‐clinical model for immune rejection studies.
Xenotransplantation – Wiley
Published: Jan 1, 2010
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