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Analysis of self-antigen specificity of islet-infiltrating T cells from human donors with type 1 diabetes

Nature Medicine volume 22, pages 1482–1487 (2016)Cite this article

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A Corrigendum to this article was published on 04 August 2017

A Corrigendum to this article was published on 07 February 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.

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Figure 1: Schema of islet handling and ex vivo isolation and growth of T cells from islets.
Figure 2: Detection of reactivity to autoreactive targets of CD4+ and CD8+ T cell lines and clones sorted or directly grown from islets from donors with T1D.
Figure 3: Detection of autoreactivity of CD4+ T cell lines and clones sorted or directly grown from islets from donors with T1D with modified peptides.

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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.

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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

Authors and 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. Department of Clinical and Experimental Medicine, Laboratory for Clinical and Experimental Endocrinology, 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, Department of Medicine, Endocrinology and Metabolism, 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

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  1. Jenny Aurielle B Babon
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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.

Corresponding author

Correspondence to Sally C Kent.

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The authors declare no competing financial interests.

Supplementary information

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. (PDF 578 kb)

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Babon, J., DeNicola, M., Blodgett, D. et al. Analysis of self-antigen specificity of islet-infiltrating T cells from human donors with type 1 diabetes. Nat Med 22, 1482–1487 (2016). https://doi.org/10.1038/nm.4203

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