Apolline Salama, 1,2,3,4 Mathilde Mosser, 1 Xavier Lévêque, 1 Andrea Perota, 5 Jean-Paul Judor, 3,4 Corentin Danna, 1 Sylvie Pogu, 1 Anne Mouré, 1 Dominique Jégou, 1 Nicolas Gaide, 6 Jérôme Abadie, 6 Olivier Gauthier, 7 Jean-Paul Concordet, 8,9,10 Stéphanie Le Bas-Bernardet, 3,4 David Riochet, 3,4,11 Ludmilla Le Berre, 3,4 Jérémy Hervouet, 3,4 David Minault, 3,4 Pierre Weiss, 12,13 Jérôme Guicheux, 12,13 Sophie Brouard, 3,4,14 Steffi Bosch, 1 Irina Lagutina, 5 Roberto Duchi, 5 Giovanna Lazzari, 5,15 Emanuele Cozzi, 16,17 Gilles Blancho, 3,4,14 Sophie Conchon, 3,4 Cesare Galli, 5,15,18 Jean-Paul Soulillou, 19 and Jean-Marie Bach 1 Neu5Gc and a1-3 GAL Xenoantigen Knockout Does Not Affect Glycemia Homeostasis and Insulin Secretion in Pigs Diabetes 2017;66:987–993 | DOI: 10.2337/db16-1060 Xenocell therapy from neonate or adult pig pancreatic islets is one of the most promising alternatives to allograft in type 1 diabetes for addressing organ short- age. In humans, however, natural and elicited anti- bodies specific for pig xenoantigens, a-(1,3)-galactose (GAL) and N-glycolylneuraminic acid (Neu5Gc), are likely to significantly contribute to xenoislet rejection. We obtained double-knockout (DKO) pigs lacking GAL and Neu5Gc. Because Neu5Gc 2/2 mice exhibit glycemic dys- regulations and pancreatic b-cell dysfunctions, we eval- uated islet function and glucose metabolism regulation in DKO pigs. Isolation of islets from neonate piglets yielded identical islet equivalent quantities to quantities obtained from control wild-type pigs. In contrast to wild-type islets, DKO islets did not induce anti-Neu5Gc antibody when grafted in cytidine monophosphate-N-acetylneuraminic acid hydroxylase KO mice and exhibited in vitro normal insulin secretion stimulated by glucose and theophylline. Adult DKO pancreata showed no histological abnormali- ties, and immunostaining of insulin and glucagon was similar to that from wild-type pancreata. Blood glucose, insulin, C-peptide, the insulin-to-glucagon ratio, and HOMA- insulin resistance in fasted adult DKO pigs and blood glucose and C-peptide changes after intravenous glu- cose or insulin administration were similar to wild-type pigs. This first evaluation of glucose homeostasis in 1 IECM, Immuno-endocrinology, EA4644 Oniris, University of Nantes, USC1383 INRA, Oniris, Nantes, France 2 Société d’Accélération du Transfert de Technologies Ouest Valorisation, Rennes, France 3 INSERM CRTI UMR 1064, University of Nantes, Nantes, France 4 Institute of Transplantation, Urology and Nephrology (ITUN), Nantes University Hospital–CHU de Nantes, Nantes, France 5 Avantea Laboratory of Reproductive Technologies, Cremona, Italy 6 Animal cancers as Models for Research in comparative Oncology (AMaROC), Oniris, Nantes, France 7 Department of Experimental Surgery, Center for Research and Preclinical Investi- gation, Oniris, Nantes, France 8 Muséum National d’Histoire Naturelle, Paris, France 9 CNRS UMR 7196, Paris, France 10 INSERM U1154, Paris, France 11 Department of Pediatrics, Nantes University Hospital –CHU de Nantes, Nantes, France 12 INSERM UMRS 791, Laboratoire d’ingénierie Ostéo-Articulaire et Dentaire (LIOAD), Nantes, France; University of Nantes, UFR Odontologie, Nantes, France 13 Nantes University Hospital–CHU de Nantes, PHU4 OTONN, Nantes, France 14 Centre d’investigation clinique (CIC) Biotherapy, Nantes University Hospital–CHU de Nantes, Nantes, France 15 Avantea Foundation, Cremona, Italy 16 Transplantation Immunology Unit, Department of Transfusion Medicine, Univer- sity of Padua–Ospedale Giustinianeo, Padua, Italy 17 CORIT (Consortium for Research in Organ Transplantation), Padua, Italy 18 Department of Veterinary Medical Sciences, University of Bologna, Ozzano Emilia, Italy 19 University of Nantes, Nantes, France Corresponding authors: Jean-Marie Bach, jean-marie.bach@oniris-nantes.fr, Jean-Paul Soulillou, soulillou@yahoo.fr, and Cesare Galli, cesaregalli@avantea.it. Received 30 August 2016 and accepted 8 January 2017. This article contains Supplementary Data online at http://diabetes .diabetesjournals.org/lookup/suppl/doi:10.2337/db16-1060/-/DC1. A.S., M.M., X.L., and A.P. contributed equally to this study. C.G., J.-P.S., and J.-M.B. contributed equally to this study. © 2017 by the American Diabetes Association. Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. More information is available at http://www.diabetesjournals .org/content/license. Diabetes Volume 66, April 2017 987 IMMUNOLOGY AND TRANSPLANTATION