IgA antibodies of coeliac disease patients recognise a dominant T cell epitope of Α-gliadin
- 1Department of Clinical Immunology, Churchill Hospital, Oxford Radcliffe Hospitals, Oxford, UK
- 2Department of Immunology, Chelsea and Westminster Hospital, London, UK
- 3The Walter and Eliza Hall Institute, Melbourne, Australia
- Correspondence to:
Dr B L Ferry
Department of Clinical Immunology, Churchill Hospital, Oxford Radcliffe Hospitals, Oxford OX3 7LJ, UK;
- Accepted 4 February 2004
- Revised 30 January 2004
Background: In coeliac disease (CD) patients, the dominant DQ2-Α-I-gliadin peptide recognised by CD4 T cells is contained within peptide sequence 57–73 (p57-73) of Α-gliadin. This peptide sequence is also located within a 33-mer protease resistant gliadin fragment and therefore is likely to play an important role in the pathogenesis of CD.
Aims: Our aim was to determine whether a B cell epitope was present within the immunodominant T cell epitope of Α-gliadin and, if so, to elucidate its sequence and determine the importance of deamidation and/or modification of the amino acid at position 65 for IgA binding.
Patients and methods: A cohort of CD patients, disease controls, and healthy individuals were examined. Serum IgA antibodies to the native and modified p57-73 fragment of Α-gliadin were analysed using enzyme linked immunosorbent assays. Peptide scanning experiments were further used to elucidate the B cell epitope.
Results and conclusion: IgA antibodies to p57-73 were found in 29/72 (40.2%) endomysial antibody positive patients, all of whom had CD. The peptide antibody appeared to be present when patients were on a diet containing gluten and declined on a gluten free diet. The p57-73 antibody was very specific for CD (98%) and had a sensitivity of 56%. The amino acid at position 65 was not important for IgA binding but was crucial for T cell recognition of p57-73. Pentapeptide PXPQP emerges as a potentially strong candidate for the IgA binding motif in this region of Α-gliadin. This study shows that a significant proportion of newly diagnosed CD patients have an antibody response to the immunodominant T cell epitope.
- CD, coeliac disease
- p57-73, Α-gliadin peptide sequence 57-73
- GFD, gluten free diet
- EMA, endomysial antibody
- Rhf, rheumatoid factor positive
- PP, paraprotein
- CV, coefficient of variation
- ELISA, enzyme linked immunosorbent assay
- TBS, Tris buffered saline
- OD, optical density