Identification of common epitopes on gliadin, enterocytes, and calreticulin recognised by antigliadin antibodies of patients with coeliac disease.
BACKGROUND
Sera of patients with coeliac disease, containing IgA and IgG antigliadin antibodies (AGA) and various IgA autoantibodies, react with isolated enterocytes. AGA cross react with enterocyte antigens, one of which has been identified as calreticulin.


OBJECTIVE
To characterise the antigenic structures of gliadin, enterocytes, and calreticulin recognised by AGA from patients with active coeliac disease.


METHODS
AGA were isolated from sera of nine patients by affinity chromatography and tested by competitive ELISA using 40 alpha-gliadin synthetic dodecapeptides (A1-F6).


RESULTS
Reactivity of gliadin with all purified AGA tested was inhibited by peptide A4 at the N-terminal region; by C2, C3, and D4 at the central region; and by F3 and F4 at the C-terminal region of the gliadin molecule. AGA cross reactivity with enterocytes was inhibited by peptides A4, D1-D4, and F6 and with calreticulin by peptides A4, D3, and D4. As dominant epitopes AGA of coeliac patients recognise similar structures corresponding to peptides A4, D3, D4, and F6 present on gliadin, enterocytes, and calreticulin. Substitution of glutamine in the A4 peptide by glutamic acid caused loss of inhibitory capacity. Shortening of peptide A4 on the N-terminal by three amino acids increased its inhibitory effect.


CONCLUSIONS
AGA of patients with coeliac disease react with similar structures on gliadin and potential autoantigens on enterocytes.
Krupicková
S
S
Department of Immunology, Institute of Microbiology, Czech Academy of Sciences, Prague.
Tucková
L
L
Flegelová
Z
Z
Michalak
M
M
Walters
J R
JR
Whelan
A
A
Harries
J
J
Vencovský
J
J
Tlaskalová-Hogenová
H
H
D000328:Adult; D000818:Animals; D000906:Antibodies / Q000097:blood; D001324:Autoantigens / Q000276:immunology; D001327:Autoimmune Diseases / Q000276:immunology; D002135:Calcium-Binding Proteins / Q000276:immunology*; D037282:Calreticulin; D002446:Celiac Disease / Q000276:immunology*; D002846:Chromatography, Affinity; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D005260:Female; D005903:Gliadin / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D007422:Intestines / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D051381:Rats; D017208:Rats, Wistar; D012261:Ribonucleoproteins / Q000276:immunology*
D000906:Antibodies; D001324:Autoantigens; D002135:Calcium-Binding Proteins; D037282:Calreticulin; D000939:Epitopes; D010446:Peptide Fragments; D012261:Ribonucleoproteins; D005903:Gliadin