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
Subscribe to Journal
Get full journal access for 1 year
only $18.75 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
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.)).
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.