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Human CD8 Responses to a Complete Epitope Set from Preproinsulin: Implications for Approaches to Epitope Discovery

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Abstract

Purpose

In this study, we explored the breadth of CD8 T cell reactivity to preproinsulin (PPI) in type 1 diabetes.

Materials and Methods

We tested a complete peptide set in pools covering all 406 potential 8–11mer epitopes of PPI and 61 algorithm-predicted human leukocyte antigen (HLA)-A2-specific epitopes (15 pools) from islet-specific glucose-6-phophatase catalytic subunit-related protein (IGRP), using a CD8-specific granzyme B enzyme-linked immunosorbent spot assay.

Results

Responses were seen to 64 of the 102 PPI pools in two or more newly diagnosed patients (63%) compared to 11 pools in the control subjects (11%, p < 0.0001, Fisher’s exact test). We identified five pools containing 20 peptides, which distinguished patients from control subjects, most of which had predicted low-affinity binding to HLA class I molecules. In contrast, fewer (5 of 15 = 33%) IGRP peptide pools, selected by higher binding affinity for HLA-A2 (present in seven of eight patients and five of seven control subjects), stimulated responses in two or more patients, and none stimulated responses in more than two control subjects (p = 0.042, Fisher’s exact test).

Conclusion

Thus, we conclude that CD8 T cell reactivity to PPI in patients with type 1 diabetes can be much broader than shown previously and more diverse than seen in control subjects. Furthermore, responses were often stimulated by peptides with low predicted HLA-binding affinities.

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Acknowledgments

C.B. was supported by a Diabetes UK studentship. F.S.W. was a Wellcome Trust Senior Fellow in Clinical Science. We are very grateful to Mr. L. Keen at the National Blood Service for the HLA typing analysis and to Mr. A.J.K. Williams and Prof. P.J. Bingley for the autoantibody measurements. We would also like to thank Mr. S.J. Chapman for his technical assistance and the physicians and diabetes specialist nurses in all of the collaborating centers for referring newly diagnosed patients to SWENDIC.

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Authors and Affiliations

  1. Department of Cellular and Molecular Medicine, School of Medical Sciences, University of Bristol, University Walk, Bristol, BS8 1TD, UK

    Caroline Baker, Alan J. Hedges & F. Susan Wong

  2. Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Bristol, UK

    Liliana G. Petrich de Marquesini, Amanda J. Bishop & Colin M. Dayan

Authors
  1. Caroline Baker
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  2. Liliana G. Petrich de Marquesini
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  3. Amanda J. Bishop
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  6. F. Susan Wong
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Corresponding author

Correspondence to F. Susan Wong.

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Baker, C., Petrich de Marquesini, L.G., Bishop, A.J. et al. Human CD8 Responses to a Complete Epitope Set from Preproinsulin: Implications for Approaches to Epitope Discovery. J Clin Immunol 28, 350–360 (2008). https://doi.org/10.1007/s10875-008-9177-4

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  • DOI: https://doi.org/10.1007/s10875-008-9177-4

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