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Gut 54:1217-1223 doi:10.1136/gut.2004.059998
  • Small intestine

T cells in peripheral blood after gluten challenge in coeliac disease

  1. R P Anderson1,
  2. D A van Heel2,
  3. J A Tye-Din1,
  4. M Barnardo3,
  5. M Salio4,
  6. D P Jewell5,
  7. A V S Hill6
  1. 1Autoimmunity and Transplantation Division, Walter and Eliza Hall Institute, and Department of Gastroenterology, the Royal Melbourne Hospital, Parkville, Victoria, Australia
  2. 2Department of Gastroenterology, Hammersmith Hospital, Imperial College, London, UK
  3. 3Transplantation Immunology, Nuffield Department of Surgery, Churchill Hospital, University of Oxford, Oxford, UK
  4. 4Weatherall Institute of Molecular Medicine, Nuffield Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford, UK
  5. 5Department of Gastroenterology, Nuffield Department of Medicine, Gibson Building, Radcliffe Infirmary, University of Oxford, Oxford, UK
  6. 6Wellcome Trust Centre for Human Genetics, Nuffield Department of Medicine, Churchill Hospital, University of Oxford, Oxford, UK
  1. Correspondence to:
    Dr R P Anderson
    Autoimmunity and Transplantation Division, Walter and Eliza Hall Institute, c/o Post Office RMH, Victoria, Australia 3050; bandersonwehi.edu.au
  • Accepted 24 February 2005
  • Revised 23 February 2005

Abstract

Background: Current understanding of T cell epitopes in coeliac disease (CD) largely derives from intestinal T cell clones in vitro. T cell clones allow identification of gluten peptides that stimulate T cells but do not quantify their contribution to the overall gluten specific T cell response in individuals with CD when exposed to gluten in vivo.

Aims: To determine the contribution of a putative dominant T cell epitope to the overall gliadin T cell response in HLA-DQ2 CD in vivo.

Patients: HLA-DQ2+ individuals with CD and healthy controls.

Methods: Subjects consumed 20 g of gluten daily for three days. Interferon γ (IFN-γ) ELISPOT was performed using peripheral blood mononuclear cells (PBMC) to enumerate and characterise peptide and gliadin specific T cells before and after gluten challenge.

Results: In 50/59 CD subjects, irrespective of homo- or heterozygosity for HLA-DQ2, IFN-γ ELISPOT responses for an optimal concentration of A-gliadin 57–73 Q-E65 were between 10 and 1500 per million PBMC, equivalent to a median 51% of the response for a “near optimal” concentration of deamidated gliadin. Whole deamidated gliadin and gliadin epitope specific T cells induced in peripheral blood expressed an intestinal homing integrin (α4β7) and were HLA-DQ2 restricted. Peripheral blood T cells specific for A-gliadin 57–73 Q-E65 are rare in untreated CD but can be predictably induced two weeks after gluten exclusion.

Conclusion: In vivo gluten challenge is a simple safe method that allows relevant T cells to be analysed and quantified in peripheral blood by ELISPOT, and should permit comprehensive high throughput mapping of gluten T cell epitopes in large numbers of individuals with CD.

Footnotes