BACKGROUND: alpha1,3-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 alpha3Gal-terminated glycolipids completely. Levels of uncapped N-acetyllactosamine precursor compounds, blood group H type 2 core chain compounds, the P1 antigen and the x(2) 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.
BACKGROUND: alpha1,3-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 pigglycolipids. 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 alpha3Gal-terminated glycolipids completely. Levels of uncapped N-acetyllactosamine precursor compounds, blood group H type 2 core chain compounds, the P1 antigen and the x(2) 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 pigglycolipids 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 pigglycolipid antigens indicating that non-human primates have limitations as a human pre-clinical model for immune rejection studies.
Authors: C Burlak; L L Paris; A J Lutz; R A Sidner; J Estrada; P Li; M Tector; A J Tector Journal: Am J Transplant Date: 2014-06-06 Impact factor: 8.086
Authors: Thomas A E Platts-Mills; Scott P Commins; Tilo Biedermann; Marianne van Hage; Michael Levin; Lisa A Beck; Maria Diuk-Wasser; Uta Jappe; Danijela Apostolovic; Michael Minnicozzi; Marshall Plaut; Jeffrey M Wilson Journal: J Allergy Clin Immunol Date: 2020-02-10 Impact factor: 10.793
Authors: Geon A Kim; Eun Mi Lee; Jun-Xue Jin; Sanghoon Lee; Anukul Taweechaipaisankul; Jong Ik Hwang; Zahid Alam; Curie Ahn; Byeong Chun Lee Journal: Transgenic Res Date: 2017-05-28 Impact factor: 2.788
Authors: Agnes M Azimzadeh; Guerard W Byrne; Mohamed Ezzelarab; Emily Welty; Gheorghe Braileanu; Xiangfei Cheng; Simon C Robson; Christopher G A McGregor; David K C Cooper; Richard N Pierson Journal: Xenotransplantation Date: 2014-09-01 Impact factor: 3.907
Authors: M Thaysen-Andersen; E Chertova; C Bergamaschi; E S X Moh; O Chertov; J Roser; R Sowder; J Bear; J Lifson; N H Packer; B K Felber; G N Pavlakis Journal: Glycoconj J Date: 2015-11-12 Impact factor: 2.916