Operational immune tolerance towards transplanted allogeneic pancreatic islets in mice and a non-human primate
Patients with autoimmune type 1 diabetes transplanted with pancreatic islets to their liver experience significant improvement in quality of life through better control of blood sugar and enhanced awareness of hypoglycaemia. However, long-term survival and efficacy of the intrahepatic islet transplant are limited owing to liver-specific complications, such as immediate blood-mediated immune reaction, hypoxia, a highly enzymatic and inflammatory environment and locally elevated levels of drugs including immunosuppressive agents, all of which are injurious to islets. This has spurred a search for new islet transplant sites and for innovative ways to achieve long-term graft survival and efficacy without life-long systemic immunosuppression and its complications.
We used our previously established approach of islet transplant in the anterior chamber of the eye in allogeneic recipient mouse models and a baboon model of diabetes, which were treated transiently with anti-CD154/CD40L blocking antibody in the peri-transplant period. Survival of the intraocular islet allografts was assessed by direct visualisation in the eye and metabolic variables (blood glucose and C-peptide measurements). We evaluated longitudinally the cytokine profile in the local microenvironment of the intraocular islet allografts, represented in aqueous humour, under conditions of immune rejection vs tolerance. We also evaluated the recall response in the periphery of the baboon recipient using delayed-type hypersensitivity (DTH) assay, and in mice after repeat transplant in the kidney following initial transplant with allogeneic islets in the eye or kidney.
Results in mice showed >300 days immunosuppression-free survival of allogeneic islets transplanted in the eye or kidney. Notably, >70% of tolerant mice, initially transplanted in the eye, exhibited >400 days of graft survival after re-transplant in the kidney without immunosuppression compared with ~30% in mice that were initially transplanted in the kidney. Cytokine and DTH data provided evidence of T helper 2-driven local and peripheral immune regulatory mechanisms in support of operational immune tolerance towards the islet allografts in both models.
We are currently evaluating the safety and efficacy of intraocular islet transplantation in a phase 1 clinical trial. In this study, we demonstrate immunosuppression-free long-term survival of intraocular islet allografts in mice and in a baboon using transient peri-transplant immune intervention. These results highlight the potential for inducing islet transplant immune tolerance through the intraocular route. Therefore, the current findings are conceptually significant and may impact markedly on clinical islet transplantation in the treatment of diabetes.
KeywordsAllogeneic rejection Anterior chamber of the eye Immune tolerance induction and maintenance Immunosuppression-free Intraocular transplantation Long-term graft survival Non-invasive longitudinal intravital imaging Pancreatic islet transplant Th2 cytokines
Immediate blood-mediated immune reaction
Peripheral blood mononuclear cell
Tetanus toxoid and diphtheria
The authors are grateful to W. Diaz, J. Geary and R. Rodriguez-Lopez (Diabetes Research Institute, University of Miami, USA) for their excellent care of non-human primates and associated procedures, and to A. Rabassa and E. Poumian-Ruiz (Diabetes Research Institute, University of Miami, USA) for assistance with islet isolation from the donor baboon. We also thank S. Dubovy and C. Maza (Bascom Palmer Eye Institute, University of Miami, USA) for help with sectioning and histological examination of the baboon eyes, and A. Mendez (Diabetes Research Institute, University of Miami, USA) and H. Salah-Uddin (Department of Psychiatry, University of Miami, USA) for help with Bio-Plex assay setup.
MHA conceived the study, designed and conducted experiments, analysed and interpreted data and wrote the manuscript. DMB designed and conducted experiments, analysed and interpreted data and wrote the manuscript. AS, CM, MH, AT, LFH, AH and EAA-Q conducted experiments and collected data and proofread the manuscript. JMP planned experiments, interpreted data and proofread the manuscript. WJB and EJ-G performed trans vivo DTH assays, interpreted data and edited the manuscript. VLP designed experiments, performed intraocular islet transplantation and eye examinations in the baboon, interpreted data and edited the manuscript. CR, NSK and P-OB conceived the study, designed experiments, interpreted data and edited the manuscript. All authors approved the version of the manuscript to be published. MHA, DMB, and P-OB are the guarantors of this work.
This work was supported by funds from the Diabetes Research Institute Foundation (DRIF) and the Diabetes Wellness Foundation and by grants from the Stanley J. Glaser Foundation Research Award (UM SJG2016-2), the NIH/NIDDK/NIAID (K01DK097194, U01-AI-102456, R56AI130330, UC4DK116241), the Swedish Diabetes Association Fund, the Swedish Research Council, Novo Nordisk Foundation, the Family Erling-Persson Foundation, Strategic Research Program in Diabetes at Karolinska Institutet, the ERC-2013-AdG 338936-BetaImage, the Family Knut and Alice Wallenberg Foundation, Skandia Insurance Company Ltd, Diabetes and Wellness Foundation, the Bert von Kantzow Foundation and the Stichting af Jochnick Foundation.
Duality of interest
P-OB is cofounder and CEO of Biocrine, an unlisted biotech company that is using the anterior chamber of the eye technique as a research tool. MHA is consultant for the same company. All other authors declare that there is no duality of interest associated with their contribution to this manuscript.
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