• A Corrigendum to this article was published on 07 February 2017
  • A Corrigendum to this article was published on 04 August 2017

This article has been updated

Abstract

A major therapeutic goal for type 1 diabetes (T1D) is to induce autoantigen-specific tolerance of T cells. This could suppress autoimmunity in those at risk for the development of T1D, as well as in those with established disease who receive islet replacement or regeneration therapy. Because functional studies of human autoreactive T cell responses have been limited largely to peripheral blood–derived T cells1,2,3, it is unclear how representative the peripheral T cell repertoire is of T cells infiltrating the islets. Our knowledge of the insulitic T cell repertoire is derived from histological and immunohistochemical analyses of insulitis4,5,6,7,8, the identification of autoreactive CD8+ T cells in situ, in islets of human leukocyte antigen (HLA)-A2+ donors9 and isolation and identification of DQ8 and DQ2–DQ8 heterodimer–restricted, proinsulin-reactive CD4+ T cells grown from islets of a single donor with T1D10. Here we present an analysis of 50 of a total of 236 CD4+ and CD8+ T cell lines grown from individual handpicked islets or clones directly sorted from handpicked, dispersed islets from nine donors with T1D. Seventeen of these T cell lines and clones reacted to a broad range of studied native islet antigens and to post-translationally modified peptides. These studies demonstrate the existence of a variety of islet-infiltrating, islet-autoantigen reactive T cells in individuals with T1D, and these data have implications for the design of successful immunotherapies.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Change history

  • 18 April 2017

    In the version of this article initially published, the IA-2545–562 peptide identified as a target of a T cell clone was mislabeled as the native version of the peptide. The T cell clone responded to a modified version of the peptide with glutamine-to-glutamic-acid deaminations at positions 548, 551 and 556. The sentence, “From donor T1D.7, a CD4+ T cell clone recognized an IA-2 peptide with three glutamine-to-glutamic-acid deaminations (IA-2545–562(Q–G548,551,556); Fig. 2e)” has been added on p.1484 to reflect this. The relevant text has also been edited in the main text on p.1483; in Figure 2e and the Figure 2 legend; and in Figure 3b. Additionally, the phrase, ‘, a deaminated IA-2 peptide’ has been added after ‘…(a panel of 60 peptides)…’ to the ‘Peptides’ section of the Online Methods. Finally, in the Figure 2 legend, the phrase ‘Detection of reactivity to known autoreactive targets…’ has been changed to ‘Detection of reactivity to autoreactive targets…’. These errors have been corrected in the HTML and PDF versions of the article.

  • 15 November 2016

    In the version of this article initially published online, the authors forgot to acknowledge the islet-isolation team at the Diabetes Research Institute, University of Miami. This oversight has been corrected for the print, PDF and HTML versions of this article.

References

  1. 1.

    , & Translational mini-review series on type 1 diabetes: Systematic analysis of T cell epitopes in autoimmune diabetes. Clin. Exp. Immunol. 148, 1–16 (2007).

  2. 2.

    Tetramer analysis of human autoreactive CD4-positive T cells. Adv. Immunol. 88, 51–71 (2005).

  3. 3.

    et al. Simultaneous detection of circulating autoreactive CD8+ T-cells specific for different islet cell-associated epitopes using combinatorial MHC multimers. Diabetes 59, 1721–1730 (2010).

  4. 4.

    et al. Blood and islet phenotypes indicate immunological heterogeneity in type 1 diabetes. Diabetes 63, 3835–3845 (2014).

  5. 5.

    et al. Insulitis and β-cell mass in the natural history of type 1 diabetes. Diabetes 65, 719–731 (2016).

  6. 6.

    et al. The diagnosis of insulitis in human type 1 diabetes. Diabetologia 56, 2541–2543 (2013).

  7. 7.

    , , , & The histopathology of the pancreas in type 1 (insulin-dependent) diabetes mellitus: a 25-year review of deaths in patients under 20 years of age in the United Kingdom. Diabetologia 29, 267–274 (1986).

  8. 8.

    Pathologic anatomy of the pancreas in juvenile diabetes mellitus. Diabetes 14, 619–633 (1965).

  9. 9.

    et al. Demonstration of islet-autoreactive CD8 T cells in insulitic lesions from recent onset and long-term type 1 diabetes patients. J. Exp. Med. 209, 51–60 (2012).

  10. 10.

    et al. Proinsulin-specific, HLA-DQ8, and HLA-DQ8-transdimer-restricted CD4+ T cells infiltrate islets in type 1 diabetes. Diabetes 64, 172–182 (2015).

  11. 11.

    et al. TCR bias of in vivo expanded T cells in pancreatic islets and spleen at the onset in human type 1 diabetes. J. Immunol. 186, 3787–3797 (2011).

  12. 12.

    et al. Evidence for superantigen involvement in insulin-dependent diabetes mellitus aetiology. Nature 371, 351–355 (1994).

  13. 13.

    et al. Mononuclear cell infiltration and its relation to the expression of major histocompatibility complex antigens and adhesion molecules in pancreas biopsy specimens from newly diagnosed insulin-dependent diabetes mellitus patients. J. Clin. Invest. 92, 2313–2322 (1993).

  14. 14.

    et al. Insulitis and characterisation of infiltrating T cells in surgical pancreatic tail resections from patients at onset of type 1 diabetes. Diabetologia 59, 492–501 (2016).

  15. 15.

    , , & The pancreas in human type 1 diabetes. Semin. Immunopathol. 33, 29–43 (2011).

  16. 16.

    et al. Recognition of self and altered self by T cells in autoimmunity and allergy. Protein Cell 4, 8–16 (2013).

  17. 17.

    , , , & Biological relevance of citrullinations: diagnostic, prognostic and therapeutic options. Autoimmunity 48, 73–79 (2015).

  18. 18.

    & Celiac disease and transglutaminase 2: a model for posttranslational modification of antigens and HLA association in the pathogenesis of autoimmune disorders. Curr. Opin. Immunol. 23, 732–738 (2011).

  19. 19.

    et al. Diabetogenic T-cell clones recognize an altered peptide of chromogranin A. Diabetes 61, 3239–3246 (2012).

  20. 20.

    et al. The insulin A-chain epitope recognized by human T cells is posttranslationally modified. J. Exp. Med. 202, 1191–1197 (2005).

  21. 21.

    , , , & T cell epitopes and post-translationally modified epitopes in type 1 diabetes. Curr. Diab. Rep. 15, 90 (2015).

  22. 22.

    et al. Citrullinated glucose-regulated protein 78 is an autoantigen in type 1 diabetes. Diabetes 64, 573–586 (2015).

  23. 23.

    et al. Antibodies to post-translationally modified insulin in type 1 diabetes. Diabetologia 58, 2851–2860 (2015).

  24. 24.

    et al. Posttranslational modification of HLA-DQ binding islet autoantigens in type 1 diabetes. Diabetes 63, 237–247 (2014).

  25. 25.

    et al. Recognition of posttranslationally modified GAD65 epitopes in subjects with type 1 diabetes. Diabetes 63, 3033–3040 (2014).

  26. 26.

    et al. Human islets and dendritic cells generate post-translationally modified islet autoantigens. Clin. Exp. Immunol. 185, 133–140 (2016).

  27. 27.

    et al. Discovery of a selective islet peptidome presented by the highest-risk HLA-DQ8trans molecule. Diabetes 65, 732–741 (2016).

  28. 28.

    et al. Pathogenic CD4 T cells in type 1 diabetes recognize epitopes formed by peptide fusion. Science 351, 711–714 (2016).

  29. 29.

    et al. Pancreatic beta cell function persists in many patients with chronic type 1 diabetes, but is not dramatically improved by prolonged immunosuppression and euglycaemia from a beta cell allograft. Diabetologia 52, 1369–1380 (2009).

  30. 30.

    , , & Recurrent diabetes mellitus in the pancreas iso- and allograft. A light and electron microscopic and immunohistochemical analysis of four cases. Lab. Invest. 53, 132–144 (1985).

  31. 31.

    et al. Recurrence of type 1 diabetes after simultaneous pancreas-kidney transplantation, despite immunosuppression, is associated with autoantibodies and pathogenic autoreactive CD4 T-cells. Diabetes 59, 947–957 (2010).

  32. 32.

    , , & Current concepts on the pathogenesis of type 1 diabetes--considerations for attempts to prevent and reverse the disease. Diabetes Care 38, 979–988 (2015).

  33. 33.

    , , & Recent lessons learned from prevention and recent-onset type 1 diabetes immunotherapy trials. Diabetes 62, 9–17 (2013).

  34. 34.

    et al. Isolation of human islets for autologous islet transplantation in children and adolescents with chronic pancreatitis. J. Transplant. 2012, 642787 (2012).

  35. 35.

    et al. Islet-enriched gene expression and glucose-induced insulin secretion in human and mouse islets. Diabetologia 55, 707–718 (2012).

  36. 36.

    et al. Novel observations from next-generation RNA sequencing of highly purified human adult and fetal islet cell subsets. Diabetes 64, 3172–3181 (2015).

  37. 37.

    et al. Residual methylprednisolone suppresses human T-cell responses to spleen, but not islet, extracts from deceased organ donors. Int. Immunol. 24, 447–453 (2012).

  38. 38.

    et al. Beta-cell replication is increased in donor organs from young patients after prolonged life support. Diabetes 59, 1702–1708 (2010).

  39. 39.

    , , , & Increased immune cell infiltration of the exocrine pancreas: a possible contribution to the pathogenesis of type 1 diabetes. Diabetes 63, 3880–3890 (2014).

  40. 40.

    et al. Protein kinase inhibitors substantially improve the physical detection of T-cells with peptide-MHC tetramers. J. Immunol. Methods 340, 11–24 (2009).

Download references

Acknowledgements

This research was performed with the support of the Network for Pancreatic Organ Donors with Diabetes (nPOD), a collaborative type 1 diabetes research project sponsored by the Juvenile Diabetes Research Foundation. Organ-procurement organizations (OPOs) partnering with nPOD to provide research resources are listed at http://www.jdrfnpod.org/for-partners/npod-partners/. We thank the families of the donors. We also thank M. Nakayama (Barbara Davis Center for Childhood Diabetes, University of Colorado) for supplying B cells from HLA-matched donors, and S. Purushothaman for her expert technical assistance. We thank D. Melton (Harvard University) for resources supporting this project. We thank G. Nepom, H. Reijonen (Benaroya Research Institute at Virginia Mason) and D. Hafler (Yale University) for providing B cell and T cell lines and clones. We also acknowledge the islet-isolation team at the Diabetes Research Institute, University of Miami. This study was supported by the University of Massachusetts Medical School Flow Cytometry Core Facility. The following funding sources supported this research: the Helmsley Charitable Trust 2015PG-T1D057 (S.C.K.), AI126189 (S.C.K.) and the Human Islet Research Network (HIRN) Opportunity Pool Fund U01 DK104162 (S.C.K.), DK089572 (A.C.P., D.M.H.), DK072473 (A.C.P.), DK104211 (A.C.P.), DK108120 (A.C.P.), DK106755 (A.C.P.), Islet Procurement and Analysis Core of the Vanderbilt Diabetes Research and Training Grant Center (DK020593) (A.C.P.), PO142288 (M.A., C. Mathews, M.C.T.), DK081166 (K.H.), Juvenile Diabetes Research Foundation 2-SRA-2015-68-Q-R (A.C.P., D.M.H.), 2-SRA-2015-52-Q-R (L.O., C. Mathieu), 2-SRA-2014-297-Q-R (E.A.J.), 25-2013-268 (M.A.A., with subcontract to J.S.K.), GOA 14/010 (L.O., C. Mathieu), American Diabetes Association Pathway to Stop Diabetes Grant 1-15-ACE-14 (T.D.), Helmsley Charitable Trust 2009PG-T1D006 (R.M.), Glass Charitable Foundation (R.M.) and the Helmsley Charitable Trust (George Eisenbarth nPOD Award for Team Science, 2015PG-T1D052 (A.P.)).

Author information

Affiliations

  1. Department of Medicine, Division of Diabetes, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, Massachusetts, USA.

    • Jenny Aurielle B Babon
    • , Megan E DeNicola
    • , David M Blodgett
    • , David M Harlan
    •  & Sally C Kent
  2. Laboratory for Clinical and Experimental Endocrinology, Department of Clinical and Experimental Medicine, KU Leuven, Leuven, Belgium.

    • Inne Crèvecoeur
    • , Lut Overbergh
    •  & Chantal Mathieu
  3. Ann Romney Center for Neurologic Diseases, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, USA.

    • Thomas S Buttrick
    •  & Wassim Elyaman
  4. Program in Molecular Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, Massachusetts, USA.

    • René Maehr
  5. Institute of Cellular Therapeutics, Allegheny-Singer Research Institute, Pittsburgh, Pennsylvania, USA.

    • Rita Bottino
  6. Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania, USA.

    • Rita Bottino
  7. Institute for Diabetes, Obesity, and Metabolism, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA.

    • Ali Naji
  8. Department of Information Sciences, Beckman Research Institute, City of Hope, Duarte, California, USA.

    • John Kaddis
  9. Benaroya Research Institute at Virginia Mason, Seattle, Washington, USA.

    • Eddie A James
  10. Division of Diabetes, Endocrinology and Metabolism, Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee, USA.

    • Rachana Haliyur
    • , Marcela Brissova
    •  & Alvin C Powers
  11. Department of Immunology and Microbiology, University of Colorado School of Medicine, Denver, Anschutz Medical Campus, Aurora, Colorado, USA.

    • Thomas Delong
    •  & Kathryn Haskins
  12. Diabetes Research Institute, University of Miami, Miami, Florida, USA.

    • Alberto Pugliese
  13. Departments of Pathology, Immunology, and Laboratory Medicine, University of Florida, Gainesville, Florida, USA.

    • Martha Campbell-Thompson
    • , Clayton Mathews
    •  & Mark A Atkinson
  14. Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, Tennessee, USA.

    • Alvin C Powers
  15. Veterans Affairs Tennessee Valley Healthcare System, Nashville, Tennessee, USA.

    • Alvin C Powers

Authors

  1. Search for Jenny Aurielle B Babon in:

  2. Search for Megan E DeNicola in:

  3. Search for David M Blodgett in:

  4. Search for Inne Crèvecoeur in:

  5. Search for Thomas S Buttrick in:

  6. Search for René Maehr in:

  7. Search for Rita Bottino in:

  8. Search for Ali Naji in:

  9. Search for John Kaddis in:

  10. Search for Wassim Elyaman in:

  11. Search for Eddie A James in:

  12. Search for Rachana Haliyur in:

  13. Search for Marcela Brissova in:

  14. Search for Lut Overbergh in:

  15. Search for Chantal Mathieu in:

  16. Search for Thomas Delong in:

  17. Search for Kathryn Haskins in:

  18. Search for Alberto Pugliese in:

  19. Search for Martha Campbell-Thompson in:

  20. Search for Clayton Mathews in:

  21. Search for Mark A Atkinson in:

  22. Search for Alvin C Powers in:

  23. Search for David M Harlan in:

  24. Search for Sally C Kent in:

Contributions

S.C.K., D.M.H. and J.A.B.B. designed the study. J.A.B.B., M.E.D., D.M.B. and S.C.K. performed experiments. T.S.B., W.E., R.H., M.B. and M.C.-T. performed experiments. R.M., I.C., E.A.J., L.O., C. Mathieu, T.D. and K.H. generated and supplied reagents. R.B., A.N., J.K., A.P., C. Mathews, M.A.A., M.B., R.H., A.C.P. and D.M.H. provided islets. S.C.K. wrote the manuscript, and all authors edited the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Sally C Kent.

Supplementary information

PDF files

  1. 1.

    Supplementary Tables and Figures

    Supplementary Tables 1–3 and Supplementary Figures 1–4 Supplemental Table 1. Summary of characteristics and tally of CD4+ and CD8+ lines and clones derived from islats of donors with T1D. Supplemental Table 2. Summary of characteristics of normal donors (Is) without T1D and lack of islet infiltrating T cells. Supplemental Table 3. Islet equivalents (IEQ) recovered from six of the islet isolations from donors with T1D. Supplemental Figure 1. One hundred and two T cells lines grown from individual islets from donors with T1D were mixtures of CD4 and CD8 T cells. Supplemental Figure 2. DQ8 restriction and proinflammatory cytokine secretion of a CD4+ T cell line reactive with hybrid peptide hEGGG:NP-Y (GQVELGGG:SSPETLI). Supplemental Figure 3. Islet-derived autoreactive T cells are pro-inflammatory. Supplemental Figure 4. Autoreactive T cell clones recognize HLA-matched B cells transduced with autoantigen.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nm.4203

Further reading