PURPOSE: Pigs expressing neither galactose-α1,3-galactose (Gal) nor N-glycolylneuraminic acid (NeuGc) take xenotransplantation one step closer to the clinic. Our aims were (1) to document the lack of NeuGc expression on corneas and aortas and cultured endothelial cells [aortic endothelial cells (AECs); corneal (CECs)] of GTKO/NeuGcKO pigs, and (2) to investigate whether the absence of NeuGc reduced human antibody binding to the tissues and cells. METHODS: Wild-type (WT), GTKO, and GTKO/NeuGcKO pigs were used for the study. Human tissues and cultured cells were negative controls. Immunofluorescence staining was performed using anti-Gal and anti-NeuGc antibodies, and human IgM and IgG binding to tissues was determined. Flow cytometric analysis was used to determine Gal and NeuGc expression on cultured CECs and AECs and to measure human IgM/IgG binding to these cells. RESULTS: Both Gal and NeuGc were detected on WT pig corneas and aortas. Although GTKO pigs expressed NeuGc, neither humans nor GTKO/NeuGcKO pigs expressed Gal or NeuGc. Human IgM/IgG binding to corneas and aortas from GTKO and GTKO/NeuGcKO pigs was reduced compared with binding to WT pigs. Human antibody binding to GTKO/NeuGcKO AECs was significantly less than that to GTKO AECs, but there was no significant difference in binding between GTKO and GTKO/NeuGcKO CECs. CONCLUSIONS: The absence of NeuGc on GTKO aortic tissue and AECs is associated with reduced human antibody binding, and possibly will provide a better outcome in clinical xenotransplantation using vascularized organs. For clinical corneal xenotransplantation, the absence of NeuGc expression on GTKO/NeuGcKO pig corneas may not prove an advantage over GTKO corneas.
PURPOSE:Pigs expressing neither galactose-α1,3-galactose (Gal) nor N-glycolylneuraminic acid (NeuGc) take xenotransplantation one step closer to the clinic. Our aims were (1) to document the lack of NeuGc expression on corneas and aortas and cultured endothelial cells [aortic endothelial cells (AECs); corneal (CECs)] of GTKO/NeuGcKO pigs, and (2) to investigate whether the absence of NeuGc reduced human antibody binding to the tissues and cells. METHODS: Wild-type (WT), GTKO, and GTKO/NeuGcKO pigs were used for the study. Human tissues and cultured cells were negative controls. Immunofluorescence staining was performed using anti-Gal and anti-NeuGc antibodies, and humanIgM and IgG binding to tissues was determined. Flow cytometric analysis was used to determine Gal and NeuGc expression on cultured CECs and AECs and to measure humanIgM/IgG binding to these cells. RESULTS: Both Gal and NeuGc were detected on WT pig corneas and aortas. Although GTKO pigs expressed NeuGc, neither humans nor GTKO/NeuGcKO pigs expressed Gal or NeuGc. HumanIgM/IgG binding to corneas and aortas from GTKO and GTKO/NeuGcKO pigs was reduced compared with binding to WT pigs. Human antibody binding to GTKO/NeuGcKO AECs was significantly less than that to GTKO AECs, but there was no significant difference in binding between GTKO and GTKO/NeuGcKO CECs. CONCLUSIONS: The absence of NeuGc on GTKO aortic tissue and AECs is associated with reduced human antibody binding, and possibly will provide a better outcome in clinical xenotransplantation using vascularized organs. For clinical corneal xenotransplantation, the absence of NeuGc expression on GTKO/NeuGcKO pig corneas may not prove an advantage over GTKO corneas.
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: Bertrand Vabres; Stéphanie Le Bas-Bernardet; David Riochet; Yan Chérel; David Minault; Jérémy Hervouet; Yvette Ducournau; Anne Moreau; Véronique Daguin; Flora Coulon; Annaïck Pallier; Sophie Brouard; Simon C Robson; Mark B Nottle; Peter J Cowan; Eric Venturi; Pascal Mermillod; Philippe Brachet; Cesare Galli; Irina Lagutina; Roberto Duchi; Jean-Marie Bach; Gilles Blancho; Jean-Paul Soulillou; Bernard Vanhove Journal: Xenotransplantation Date: 2014-07-08 Impact factor: 3.907
Authors: Annie N Samraj; Oliver M T Pearce; Heinz Läubli; Alyssa N Crittenden; Anne K Bergfeld; Kalyan Banda; Christopher J Gregg; Andrea E Bingman; Patrick Secrest; Sandra L Diaz; Nissi M Varki; Ajit Varki Journal: Proc Natl Acad Sci U S A Date: 2014-12-29 Impact factor: 11.205
Authors: Minoru Fujita; Ruhina Mehra; Seung Eun Lee; Danny S Roh; Cassandra Long; James L Funderburgh; David L Ayares; David K C Cooper; Hidetaka Hara Journal: Ophthalmic Res Date: 2012-12-18 Impact factor: 2.892
Authors: Christopher Burlak; Marshall Bern; Alejandro E Brito; Dragan Isailovic; Zheng-Yu Wang; Jose L Estrada; Ping Li; A Joseph Tector Journal: Xenotransplantation Date: 2013-09-05 Impact factor: 3.907
Authors: Seung Eun Lee; Ruhina Mehra; Minoru Fujita; Danny S Roh; Cassandra Long; Whayoung Lee; James L Funderburgh; David L Ayares; David K C Cooper; Hidetaka Hara Journal: Semin Ophthalmol Date: 2013-06-12 Impact factor: 1.975
Authors: Guerard W Byrne; Zeji Du; Paul Stalboerger; Heide Kogelberg; Christopher G A McGregor Journal: Xenotransplantation Date: 2014-09-01 Impact factor: 3.907
Authors: Whayoung Lee; Hidetaka Hara; Mohamed B Ezzelarab; Hayato Iwase; Rita Bottino; Cassandra Long; Jagdeece Ramsoondar; David Ayares; David K C Cooper Journal: Xenotransplantation Date: 2016-03-14 Impact factor: 3.907
Authors: Hironosuke Watanabe; Yuichi Ariyoshi; Thomas Pomposelli; Kazuhiro Takeuchi; Dilrukshi K Ekanayake-Alper; Lennan K Boyd; Scott J Arn; Hisashi Sahara; Akira Shimizu; David Ayares; Marc I Lorber; Megan Sykes; David H Sachs; Kazuhiko Yamada Journal: Xenotransplantation Date: 2019-09-23 Impact factor: 3.907