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Gut 46:46-51 doi:10.1136/gut.46.1.46
  • Small intestine

Production of a panel of recombinant gliadins for the characterisation of T cell reactivity in coeliac disease

  1. E H Arentz-Hansen,
  2. S N McAdam,
  3. Ø Molberg,
  4. C Kristiansen,
  5. L M Sollid
  1. Institute of Immunology, Rikshospitalet, University of Oslo, Norway
  1. Dr E H Arentz-Hansen, Institute of Immunology, Rikshospitalet, 0027 Oslo, Norway.
  • Accepted 21 July 1999

Abstract

BACKGROUND/AIMS Coeliac disease is a chronic intestinal disorder most probably caused by an abnormal immune reaction to wheat gliadin. The identification of the HLA-DQ2 and HLA-DQ8 as the molecules responsible for the HLA association in coeliac disease strongly implicates a role for CD4 T cells in disease pathogenesis. Indeed, CD4 T cells specific for gliadin have been isolated from the small intestine of patients with coeliac disease. However, identification of T cell epitopes within gliadin has been hampered by the heterogeneous nature of the gliadin antigen. To aid the characterisation of gliadin T cell epitopes, multiple recombinant gliadins have been produced from a commercial Nordic wheat cultivar.

METHODS The α-gliadin and γ-gliadin genes were amplified by polymerase chain reaction from cDNA and genomic DNA, cloned into a pET expression vector, and sequenced. Genes encoding mature gliadins were expressed inEscherichia coli and tested for recognition by T cells.

RESULTS In total, 16 α-gliadin genes with complete open reading frames were sequenced. These genes encoded 11 distinct gliadin proteins, only one of which was found in the Swiss-Prot database. Expression of these gliadin genes produced a panel of recombinant α-gliadin proteins of purity suitable for use as an antigen for T cell stimulation.

CONCLUSION This study provides an insight into the complexity of the gliadin antigen present in a wheat strain and has defined a panel of pure gliadin antigens that should prove invaluable for the future mapping of epitopes recognised by intestinal T cells in coeliac disease.

Footnotes

  • Abbreviations used in this paper:
    PCR
    polymerase chain reaction
    SDS/PAGE
    sodium dodecyl sulphate/polyacrylamide gel electrophoresis
    tTG
    tissue transglutaminase
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