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Avenin fails to induce a Th1 response in coeliac tissue following in vitro culture
  1. C Kilmartin1,
  2. S Lynch1,
  3. M Abuzakouk1,
  4. H Wieser2,
  5. C Feighery1
  1. 1C Kilmartin, S Lynch, M Abuzakouk, C Feighery, Department of Immunology, St James's Hospital, James's Street, Dublin 8, Ireland
  2. 2H Wieser, Deutsche Forschungsanstalt, für Lebensmittelchemie, Lichtenbergstr 4, D-85748 Garching, Germany
  1. Correspondence to:
    C Kilmartin Department of Immunology, St James's Hospital, James's Street, Dublin 8, Ireland;
    kilmarc{at}tcd.ie

Abstract

Background: It is well established that the wheat protein gliadin triggers inflammation in coeliac patients. However, the potential toxicity of avenin, the equivalent protein in oats, is debated.

Aim: To investigate the immunogenicity of avenin using the cytokines interferon γ (IFN-γ) and interleukin (IL)-2 as markers of immunological activity.

Methods: Duodenal biopsies from coeliac patients were cultured with 5 mg/ml of peptic tryptic (PT) gliadin (n=9) or 5 mg/ml of PT avenin (n=8) for four hours. Biopsies cultured with RPMI 1640 alone served as controls. Non-coeliac biopsies were also cultured with PT gliadin (n=8) and PT avenin (n=8). Total RNA was extracted from the tissue after culture. Cytokine mRNA was quantified by TaqMan polymerase chain reaction. Secreted cytokine protein was measured in the culture supernatant by enzyme linked immunosorbent assay.

Results: After culture with PT gliadin, an increase in IFN-γ mRNA was observed in all nine patients with coeliac disease. Increased IFN-γ protein was also found in four of these patients. Smaller increases in IL-2 mRNA were detected in six subjects with increased IL-2 protein found in two patients. In contrast with PT gliadin, there was no significant IFN-γ or IL-2 response when coeliac biopsies were cultured with PT avenin. Similarly, biopsies from normal controls did not respond to PT gliadin or PT avenin stimulation.

Conclusions: The findings of this study suggest that the immunogenic sequences in gliadin are not present in avenin. Moreover, they are in keeping with in vivo studies which report that oats are safe for consumption by coeliac patients.

  • coeliac disease
  • avenin
  • organ culture
  • interferon γ
  • interleukin 2
  • IFN-γ, interferon γ
  • IL, interleukin
  • PT, peptic tryptic
  • PCR, polymerase chain reaction
  • ELISA, enzyme linked immunosorbent assay
  • IEL, intraepithelial lymphocyte
  • cDNA, complementary DNA
  • GAPDH, glyceraldehyde-3-phosphate dehydrogenase
  • Tm, melting temperature
  • PBMC, peripheral blood mononuclear cell
  • EMA, endomysial antibody
  • ICAM, intercellular adhesion molecule
  • RT, room temperature (≈20°C)
  • RP-HPLC, reversed phase-high performance liquid chromatography
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