1. Production of a panel of recombinant gliadins for the characterisation of T cell reactivity in coeliac disease.
- Author
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Arentz-Hansen EH, McAdam SN, Molberg O, Kristiansen C, and Sollid LM
- Subjects
- Amino Acid Sequence, Electrophoresis, Polyacrylamide Gel, Epitope Mapping methods, Gliadin genetics, Gliadin immunology, Humans, Intestine, Small immunology, Molecular Sequence Data, Polymerase Chain Reaction, Recombinant Proteins biosynthesis, Celiac Disease immunology, Gliadin biosynthesis, T-Lymphocyte Subsets immunology
- 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 alpha-gliadin and gamma-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 in Escherichia coli and tested for recognition by T cells., Results: In total, 16 alpha-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 alpha-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.
- Published
- 2000
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