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In vivo antigen challenge in celiac disease identifies a single transglutaminase-modified peptide as the dominant A-gliadin T-cell epitope

Abstract

Celiac disease (CD) is an increasingly diagnosed enteropathy (prevalence, 1:200–1:300)1 that is induced by dietary exposure to wheat gliadins2 (as well as related proteins in rye and barley) and is strongly associated with HLA-DQ2 (α1*0501, β1*0201), which is present in over 90% of CD patients3. Because a variety of gliadin peptides have been identified as epitopes for gliadin-specific T-cell clones4,5,6 and as bioactive sequences in feeding studies and in ex vivo CD intestinal biopsy challenge7,8,9, it has been unclear whether a ‘dominant’ T-cell epitope is associated with CD. Here, we used fresh peripheral blood lymphocytes from individual subjects undergoing short-term antigen challenge and tissue transglutaminase-treated, overlapping synthetic peptides spanning A-gliadin to demonstrate a transient, disease-specific, DQ2-restricted, CD4 T-cell response to a single dominant epitope. Optimal gamma interferon release in an ELISPOT assay was elicited by a 17-amino-acid peptide corresponding to the partially deamidated peptide of A-gliadin amino acids 57–73 (Q65E). Consistent with earlier reports indicating that host tissue transglutaminase modification of gliadin enhances gliadin-specific CD T-cell responses10, tissue transglutaminase specifically deamidated Q65 in the peptide of A-gliadin amino acids 56–75. Discovery of this dominant epitope may allow development of antigen-specific immunotherapy for CD.

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Figure 1: Amino-acid sequence of A-gliadin
Figure 2: Ex vivo IFN-γ responses of PBMCs to A-gliaden peptides.
Figure 3: Definition of the minimal epitope in tTG-treated pool 3.
Figure 4: Analysis of dominant A-gliadin epitope.
Figure 5: Dynamics of PBMC IFN-γ responses by ELISPOT assay and proliferative responses.

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Acknowledgements

We thank A. Willis for HPLC and amino-acid analysis; G. Bird and H. Griffith for assays of antibody against endomysium; M. Bunce for tissue typing; A. Pathan and A. Lalvani for the gift of antibodies against class II; A. Connell for dietary advice; and J. Simmons and A. Ellis for referring subjects for the study. R.P.A. was supported by a CJ Martin/Menzies Fellowship from the National Health and Medical Research Council of Australia 987001. A.V.S.H. is a Wellcome Trust Principal Research Fellow. The work was supported by the Coeliac Society of the United Kingdom and Isis Innovation (University of Oxford, UK).

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Correspondence to Robert P. Anderson.

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Anderson, R., Degano, P., Godkin, A. et al. In vivo antigen challenge in celiac disease identifies a single transglutaminase-modified peptide as the dominant A-gliadin T-cell epitope. Nat Med 6, 337–342 (2000). https://doi.org/10.1038/73200

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