title: The toxicity of high molecular weight glutenin subunits of wheat to patients with coeliac disease.
abstract: OBJECTIVE
  The ability of the gliadin fraction of wheat gluten to exacerbate coeliac disease is well documented. We investigated the possible toxicity of high molecular weight glutenin subunits (HMW-GS) in coeliac disease in vitro using gluten-sensitive T cells, and in vivo with challenge studies in patient volunteers.
  
  
  METHODS
  A mixture of four HMW-GS was chemically separated from wheat flour and checked for purity by HPLC, SDS-PAGE and ELISA. T-cell lines, grown up from small intestinal biopsies from coeliac patients (n=17), were tested for their reactivity to HMW-GS. Adults with coeliac disease and who were on a gluten-free diet (n=3) underwent in-vivo challenges with HMW-GS. Duodenal biopsies, taken prior to the challenge and at intervals up to 6 h afterwards, were assessed for morphology, intra-epithelial lymphocyte count, and interleukin 15 (IL-15) expression, by immunohistochemistry.
  
  
  RESULTS
  T-cell lines from 11 of 17 patients were stimulated by HMW-GS. There was a significant change in small intestinal morphology 4 h after commencing infusions with HMW-GS in all three subjects. For example villus height to crypt depth ratios were reduced in the three patients from 3.0+/-0.5 to 1.29+/-0.2, 2.53+/-0.7 to 0.81+/-0.6 and 3.0+/-0.7 to 1.85+/-0.3, P<0.0001 in all cases. There was increased expression of IL-15 in the small intestine from 2 h after the HMW-GS challenges.
  
  
  CONCLUSIONS
  Mixed HMW-GS stimulate T-cell lines from some coeliac patients and exacerbate coeliac disease in vivo, inducing expression, within 2 h, of IL-15. This suggests an innate immune response to these proteins.
authors.affiliation: Rayne Institute (KCL Division of Nutrition), St Thomas' Hospital, Lambeth Palace Road, London, UK.
authors.forename: David H
authors.initials: DH
authors.lastname: Dewar
authors.forename: Massimo
authors.initials: M
authors.lastname: Amato
authors.forename: H Julia
authors.initials: HJ
authors.lastname: Ellis
authors.forename: Emma L
authors.initials: EL
authors.lastname: Pollock
authors.forename: Nuria
authors.initials: N
authors.lastname: Gonzalez-Cinca
authors.forename: Herbert
authors.initials: H
authors.lastname: Wieser
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Ciclitira
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002446:Celiac Disease / Q000276:immunology / Q000473:pathology*; D002460:Cell Line; D018753:Diet, Protein-Restricted; D000939:Epitopes / Q000276:immunology; D005260:Female; D005983:Glutens / Q000008:administration & dosage / Q000009:adverse effects* / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007371:Interferon-gamma / Q000276:immunology; D019409:Interleukin-15 / Q000032:analysis; D007413:Intestinal Mucosa / Q000276:immunology / Q000473:pathology; D007421:Intestine, Small / Q000276:immunology / Q000473:pathology; D008297:Male; D008875:Middle Aged; D008970:Molecular Weight; D013601:T-Lymphocytes / Q000276:immunology; D014908:Triticum / Q000737:chemistry*
chemical_list: D000939:Epitopes; D019409:Interleukin-15; D005983:Glutens; D007371:Interferon-gamma; C037302:glutenin