Journal of Clinical Immunology

, Volume 28, Issue 4, pp 350–360

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

  • Caroline Baker
  • Liliana G. Petrich de Marquesini
  • Amanda J. Bishop
  • Alan J. Hedges
  • Colin M. Dayan
  • F. Susan Wong
Article

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.

Keywords

Type 1 diabetes autoimmunity human CD8 T cells 

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Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Caroline Baker
    • 1
  • Liliana G. Petrich de Marquesini
    • 2
  • Amanda J. Bishop
    • 2
  • Alan J. Hedges
    • 1
  • Colin M. Dayan
    • 2
  • F. Susan Wong
    • 1
  1. 1.Department of Cellular and Molecular Medicine, School of Medical SciencesUniversity of BristolBristolUK
  2. 2.Henry Wellcome Laboratories for Integrative Neuroscience and EndocrinologyUniversity of BristolBristolUK

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