In vivo imaging of type 1 diabetes immunopathology using eye-transplanted islets in NOD mice
Autoimmune attack against the insulin-producing beta cells in the pancreatic islets results in type 1 diabetes. However, despite considerable research, details of the type 1 diabetes immunopathology in situ are not fully understood mainly because of difficult access to the pancreatic islets in vivo.
Here, we used direct non-invasive confocal imaging of islets transplanted in the anterior chamber of the eye (ACE) to investigate the anti-islet autoimmunity in NOD mice before, during and after diabetes onset. ACE-transplanted islets allowed longitudinal studies of the autoimmune attack against islets and revealed the infiltration kinetics and in situ motility dynamics of fluorescence-labelled autoreactive T cells during diabetes development. Ex vivo immunostaining was also used to compare immune cell infiltrations into islet grafts in the eye and kidney as well as in pancreatic islets of the same diabetic NOD mice.
We found similar immune infiltration in native pancreatic and ACE-transplanted islets, which established the ACE-transplanted islets as reliable reporters of the autoimmune response. Longitudinal studies in ACE-transplanted islets identified in vivo hallmarks of islet inflammation that concurred with early immune infiltration of the islets and preceded their collapse and hyperglycaemia onset. A model incorporating data on ACE-transplanted islet degranulation and swelling allowed early prediction of the autoimmune attack in the pancreas and prompted treatments to intercept type 1 diabetes.
The current findings highlight the value of ACE-transplanted islets in studying early type 1 diabetes pathogenesis in vivo and underscore the need for timely intervention to halt disease progression.
KeywordsAnterior chamber of the eye Autoimmune diabetes Diabetes recurrence Diabetes transfer Immune modulation Intraocular transplantation Islet degranulation Islet inflammation Islet swelling Local intervention NOD mice Non-invasive longitudinal intravital imaging Pancreatic islet transplant Prediction of type 1 diabetes Predictive mathematical model
Anterior chamber of the eye
In vivo immunocytolabelling
Median fluorescence intensity
Median time to hyperglycaemia
Positron emission tomography
The authors are grateful to R. Rodriguez-Diaz (DRI, University of Miami, USA) and I. Leibiger (The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Stockholm, Sweden) for fruitful discussion of the manuscript. Special thanks go to E. Zahar-Akrawi, J. Gimeno and Y. Gadea (DRI Translational Core, University of Miami, USA), O. Umland (DRI Flow Cytometry Core, University of Miami, USA) and K. Johnson (DRI Histology Core, University of Miami, USA) for outstanding technical assistance.
MHA, RDM and AP conceived and designed the study, conducted experiments, analysed and interpreted data and wrote the manuscript. GF, ASB and CF planned experiments, analysed and interpreted data and edited the manuscript. MLC, UU, CF, AS, LFH, AT, VA and AT-G, conducted experiments, collected data and proof-read the manuscript. PB planned experiments, analysed and interpreted data, developed the mathematical model and wrote the manuscript. CR contributed to discussion and advice on experimental design and reviewed the manuscript. AC 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, RDM, PB, AP and P-OB are the guarantors of this work. The contents of this manuscript are solely the responsibility of the authors and do not necessarily represent the official views of the funding agencies. This article was prepared while AP was employed at the University of Miami. He is currently employed at NIH/Center for Scientific Review. The opinions expressed in this article are the authors’ own and do not reflect the view of the NIH, the Department of Health and Human Services or the United States government.
This work was supported by funds from the Diabetes Research Institute Foundation (DRIF; to MHA, AC, PB, P-OB and AP), Diabetes Research & Wellness Foundation and Diabetes Wellness Sverige (to MHA and P-OB), the National Institutes of Health (NIH), the National Institute of Allergy and Infectious Diseases (NIAID) - Cooperative Study Group for Autoimmune Disease Prevention U19AI050864 (to AP), R56AI130330 (to MHA), the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) R03DK075487 (to AC), R01DK084321 (to AC), UC4DK116241/K01DK097194/ F32DK083226 (to MHA); the Juvenile Diabetes Research Foundation International (JDRF) 4-2004-361 (to AC, P-OB and AP) and 4-2008-811 and 17-2010-5 (to AP). Additional support to P-OB was received from 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 European Research Council (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 ACE 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.
- 14.Sabek O, Gaber MW, Wilson CM, Zawaski JA, Fraga DW, Gaber O (2010) Imaging of human islet vascularization using a dorsal window model. Transplant Proc 42(6):2112–2114. https://doi.org/10.1016/j.transproceed.2010.05.080 CrossRefPubMedGoogle Scholar
- 15.Takahashi Y, Takebe T, Enomura M et al (2014) High-resolution intravital imaging for monitoring the transplanted islets in mice. Transplant Proc 46(4):1166–1168. https://doi.org/10.1016/j.transproceed.2013.11.089 CrossRefPubMedGoogle Scholar
- 16.Krishnan R, Arora RP, Alexander M et al (2014) Noninvasive evaluation of the vascular response to transplantation of alginate encapsulated islets using the dorsal skin-fold model. Biomaterials 35(3):891–898. https://doi.org/10.1016/j.biomaterials.2013.10.012 CrossRefPubMedGoogle Scholar
- 17.Benson RA, Garcon F, Recino A et al (2018) Non-invasive multiphoton imaging of islets transplanted into the pinna of the NOD mouse ear reveals the immediate effect of anti-CD3 treatment in autoimmune diabetes. Front Immunol 9:1006. https://doi.org/10.3389/fimmu.2018.01006 CrossRefPubMedPubMedCentralGoogle Scholar
- 18.Abdulreda MH, Caicedo A, Berggren P-O (2013) Transplantation into the anterior chamber of the eye for longitudinal, non-invasive in vivo imaging with single-cell resolution in real-time. J Vis Exp (73):e50466. https://doi.org/10.3791/50466
- 30.Chen Y, Chong MM, Darwiche R, Thomas HE, Kay TW (2004) Severe pancreatitis with exocrine destruction and increased islet neogenesis in mice with suppressor of cytokine signaling-1 deficiency. Am J Pathol 165(3):913–921. https://doi.org/10.1016/S0002-9440(10)63353-6 CrossRefPubMedPubMedCentralGoogle Scholar
- 32.Wan X, Guloglu FB, Vanmorlan AM et al (2013) Recovery from overt type 1 diabetes ensues when immune tolerance and beta-cell formation are coupled with regeneration of endothelial cells in the pancreatic islets. Diabetes 62(8):2879–2889. https://doi.org/10.2337/db12-1281 CrossRefPubMedPubMedCentralGoogle Scholar
- 33.Bodor N, Buchwald P (2000) Soft drug design: general principles and recent applications. Med Res Rev 20(1):58–101. https://doi.org/10.1002/(SICI)1098-1128(200001)20:1<58::AID-MED3>3.0.CO;2-X CrossRefPubMedGoogle Scholar
- 50.Pileggi AAM, Faleo G, Molano RD, Ricordi C, Caicedo A, Berggren P-O (2012) The anterior chamber of the eye allows longitudinal, intravital imaging of CTLs to study islet immunobiology [Eletter]. J Clin Invest. https://doi.org/10.1172/JCI59285L1
- 52.Viret C, Mahiddine K, Baker RL, Haskins K, Guerder S (2015) The T cell repertoire-diversifying enzyme TSSP contributes to thymic selection of diabetogenic CD4 T cell specificities reactive to ChgA and IAPP autoantigens. J Immunol 195(5):1964–1973. https://doi.org/10.4049/jimmunol.1401683 CrossRefPubMedGoogle Scholar
- 59.Vendrame F, Pileggi A, Laughlin E et al (2010) Recurrence of type 1 diabetes after simultaneous pancreas-kidney transplantation, despite immunosuppression, is associated with autoantibodies and pathogenic autoreactive CD4 T-cells. Diabetes 59(4):947–957. https://doi.org/10.2337/db09-0498 CrossRefPubMedPubMedCentralGoogle Scholar