SMALL INTESTINE

T cells in peripheral blood after gluten challenge in coeliac disease

R P Anderson¹, D A van Heel², J A Tye-Din¹, M Barnardo³, M Salio⁴, D P Jewell⁵, A V S Hill⁶

¹ Autoimmunity and Transplantation Division, Walter and Eliza Hall Institute, and Department of Gastroenterology, the Royal Melbourne Hospital, Parkville, Victoria, Australia
² Department of Gastroenterology, Hammersmith Hospital, Imperial College, London, UK
³ Transplantation Immunology, Nuffield Department of Surgery, Churchill Hospital, University of Oxford, Oxford, UK
⁴ Weatherall Institute of Molecular Medicine, Nuffield Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford, UK
⁵ Department of Gastroenterology, Nuffield Department of Medicine, Gibson Building, Radcliffe Infirmary, University of Oxford, Oxford, UK
⁶ Wellcome Trust Centre for Human Genetics, Nuffield Department of Medicine, Churchill Hospital, University of Oxford, Oxford, UK

Correspondence to:
Correspondence to:
Dr R P Anderson
Autoimmunity and Transplantation Division, Walter and Eliza Hall Institute, c/o Post Office RMH, Victoria, Australia 3050; banderson{at}wehi.edu.au

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 (IFN-) 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- 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 (4ß7) 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.

Abbreviations: CD, coeliac disease; IFN-, interferon ; IL, interleukin; GFD, gluten free diet; PBMC, peripheral blood mononuclear cells; tTG, tissue transglutaminase; EMA, endomysial antibodies; PPD, purified protein derivative of Mycobacterium bovis; SFU, spot forming units

Keywords: coeliac disease; T cell epitopes; gluten; interferon ; ELISPOT; antigen challenge

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