Deamidated gliadin peptides form epitopes that transglutaminase antibodies recognize.
OBJECTIVE
Deamidated gliadin peptides are efficient antigens in diagnostic tests for celiac disease, and results correlate better with transglutaminase 2-based assays than those with native gliadin. We investigated whether deamidated gliadin antigens are structurally similar to transglutaminase 2 or could mimic transglutaminase epitopes.


METHODS
Serum samples from 74 celiac and 65 control patients, and 13 different transglutaminase 2-specific monoclonal mouse antibodies were investigated for their binding to commercially available deamidated gliadin peptides using enzyme-linked immunosorbent assay, competition studies, and molecular modelling.


RESULTS
The enzyme-linked immunosorbent assay with deamidated gliadin peptides had 100% sensitivity and 98.5% specificity in patients. Deamidated gliadin epitopes also were recognized by 3 transglutaminase-specific monoclonal antibodies, and antibodies affinity-purified with deamidated gliadin peptides from celiac patient sera reacted with transglutaminase but did not show endomysial binding. The binding of the monoclonal antibodies to deamidated gliadin was inhibited dose dependently by full-length recombinant human transglutaminase, its fragments containing the binding sites of these monoclonal antibodies, or by celiac patient antibodies. Deamidated gliadin peptides decreased the binding of transglutaminase-specific monoclonal antibodies to transglutaminase. Three different cross-reacting transglutaminase epitopes were found, of which 2 are located in the C-terminal domain and 1 is conformational. The binding of celiac serum samples to deamidated gliadin peptides could not be abolished by transglutaminase or by any of the transglutaminase-specific monoclonals, indicating that celiac sera also contain additional antibodies to gliadin epitopes different from transglutaminase.


CONCLUSIONS
Certain deamidated gliadin-derived peptides and transglutaminase 2 epitopes have similar 3-dimensional appearance. This homology may contribute to the induction of transglutaminase autoantibodies by molecular mimicry.
Korponay-Szabó
Ilma R
IR
Departments of Pediatrics, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary.
Vecsei
Zsófia
Z
Király
Róbert
R
Dahlbom
Ingrid
I
Chirdo
Fernando
F
Nemes
Eva
E
Fésüs
László
L
Mäki
Markku
M
D000293:Adolescent; D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal; D016022:Case-Control Studies; D002446:Celiac Disease / Q000276:immunology*; D002648:Child; D002675:Child, Preschool; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D000939:Epitopes; D005260:Female; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000276:immunology*; D005903:Gliadin / Q000737:chemistry / Q000276:immunology*; D005983:Glutens / Q000008:administration & dosage; D006801:Humans; D007223:Infant; D008297:Male; D051379:Mice; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology*; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D011503:Transglutaminases / Q000737:chemistry / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010446:Peptide Fragments; D005983:Glutens; D005903:Gliadin; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins