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ResearchIn-Press PreviewImmunology Open Access | 10.1172/jci.insight.203119

Tracking GAD-specific T-cell expansions in Type 1 diabetes by intradermal GAD-Alum challenge

Stephanie J. Hanna,1 Emma J.S. Robinson,1 Terri C. Thayer,1 Maki Nakayama,2 Laurie Landry,2 Robert Andrews,1 Garry Dolton,1 Joanne Davies,1 Evangelia Williams,3 James A. Pearson,4 Andrew K. Sewell,1 Parth Narendran,4 David Wraith,4 Alexandra Howell,1 Philippa Young,5 Mary Hart,6 Anton Lindqvist,7 F. Susan Wong,1 Tim I.M. Tree,3 Colin M. Dayan,4 and Danijela Tatovic4

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Hanna, S. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Robinson, E. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Thayer, T. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Nakayama, M. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Landry, L. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Andrews, R. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Dolton, G. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Davies, J. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Williams, E. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Pearson, J. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Sewell, A. in: PubMed | Google Scholar |

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Narendran, P. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Wraith, D. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Howell, A. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Young, P. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Hart, M. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Lindqvist, A. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Wong, F. in: PubMed | Google Scholar |

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Tree, T. in: PubMed | Google Scholar

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Dayan, C. in: PubMed | Google Scholar |

1Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom

2Barbara Davis Center for Childhood Diabetes, University of Colorado, Denver, United States of America

3Department of Immunobiology, School of Immunology & Microbial Sciences, King’s College London, London, United Kingdom

4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom

5Breast Test Wales, Public Health Wales, Cardiff, United Kingdom

6Department of Radiology, University Hospital of Wales, Cardiff, United Kingdom

7Diamyd Medical AB, Stockholm, Sweden

Find articles by Tatovic, D. in: PubMed | Google Scholar

Published September 1, 2026 - More info

JCI Insight. https://doi.org/10.1172/jci.insight.203119.
Copyright © 2026, Hanna et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Published September 1, 2026 - Version history
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Abstract

Identifying and monitoring autoreactive T cells that drive beta cell destruction remains a major obstacle to developing effective immunotherapies for type 1 diabetes (T1D). These cells are extremely rare in peripheral blood and cannot be accessed directly from the pancreas. We used intradermal injection of Glutamic Acid Decarboxylase (GAD)-Alum to recruit GAD-specific T cells to accessible sites in the skin and skin-draining lymph nodes (LNs), sampled by skin suction blisters and ultrasound-guided LN aspiration. Peripheral blood samples obtained before GAD injection were restimulated with GAD in vitro to detect reactive CD4+ T cells. Single-cell RNA sequencing (scRNAseq) followed by re-expression of selected T cell receptors (TCRs) confirmed antigen specificity. Up to 70% of T cells at the skin injection site were clonally-expanded and 4 of 14 (28%) re-expressed TCRs were GAD-reactive. In LNs 1 of 14 (4%) clonally-expanded TCRs was GAD-reactive, representing ~0.08% of all T-cells. GAD-reactive cells across compartments displayed Th1 and Th17-associated transcription signatures. These results demonstrate the intradermal autoantigen challenge and scRNAseq, enable direct identification and molecular profiling of autoreactive T cells in vivo. This minimally invasive approach provides a powerful platform for tracking antigen-specific T cells to monitor disease activity and evaluate immune interventions in T1D.

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