Abstract

BACKGROUND
Wheat gluten comprises gliadins and glutenins. The high-molecular-weight (HMW) glutenin subunits (GS)-1Dy10 are toxic for patients with celiac disease (CD). This study aimed to assess whether CD patients mount a serological response to HMW-GS-1Dy10.
METHODS
Recombinant HMW-GS-1Dy10 was deamidated using human recombinant tissue transglutaminase. MALDI-TOF was performed to compare the level of deamidation of glutamine residues between material before and after treatment. Enzyme-linked immunosorbent assays were developed. Sera from patients with untreated CD and gastrointestinal disease controls were tested and receiver operator characteristics were used to calculate cutoffs.
RESULTS
MALDI-TOF revealed a number of fragments matching known HMW-GS-1Dy10 sequences within both the deamidated and non-deamidated material. Evidence of deamidation of glutamine residues was found only within the human transglutaminase-treated material. Patients with untreated CD had significantly increased levels of serum antibodies to HMW-GS-1Dy10 compared to controls. Undeamidated HMW-GS-1Dy10 IgA antibodies had sensitivities and specificities of 72.5 and 78.26%, respectively. Deamidated HMW-GS-1Dy10 IgA antibodies had sensitivities and specificities of 76.8 and 65.2%. Undeamidated HMW-GS-1Dy10 IgG antibodies had sensitivities and specificities of 75.3 and 68.1%. Deamidated HMW-GS-1Dy10 IgG antibodies had sensitivities and specificities of 36.2 and 92.8%.
CONCLUSION
Patients with untreated CD have raised antibody levels to HMW-GS-1Dy10, indicating the participation of these proteins in the adaptive immune response to gluten. Discrimination between CD patients and controls is not enhanced by deamidation of HMW-GS-1Dy10. Thus antibodies to these proteins are not useful markers for CD detection.

Links

Publisher Full Text

Authors

Ellis HJ, Lozano-Sanchez P, Bermudo Redondo C, Šuligoj T, Biagi F, Bianchi PI, Corazza GR, De Silvestri A, Bravi E, Katakis I, O'Sullivan CK, Ciclitira PJ

Institution

Division of Diabetes and Nutritional Sciences, King's College London, London, UK. julia.ellis@kcl.ac.uk

Source

International archives of allergy and immunology 159:4 2012 pg 428-34

MeSH

Adaptive Immunity
Amino Acid Sequence
Antibodies
Antibody Specificity
Celiac Disease
Gliadin
Glutens
Humans
Immunoglobulin A
Molecular Sequence Data
Molecular Weight
Protein Subunits
Recombinant Proteins
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Transglutaminases
Triticum

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22813868