Fine specificity of monoclonal antibodies against celiac disease-inducing peptides in the gluteome.
BACKGROUND
In celiac disease patients, peptides derived from dietary gluten are recognized by HLA-DQ2-restricted CD4(+) T cells, which results in inflammation. Such immune-stimulatory peptides are found in both gliadins and glutenins. Monoclonal antibodies (mAbs) against these peptides can be used to screen food for the presence of such peptides.


OBJECTIVE
We aimed to determine the specificity of 5 mAbs raised against T cell stimulatory peptides found in alpha- and gamma-gliadins and in low- and high-molecular-weight glutenins and to compare it with the specificity of patient-derived T cells.


METHODS
The reactivity of the mAbs with gluten peptides, enzymatic gluten digests, and intact gluten proteins was determined and compared with that of gluten-specific T cells by using a combination of immunologic and biochemical techniques. Furthermore, the reactivity of the mAbs with gluten homologues in barley, rye, and oat was determined.


RESULTS
The specificity of the mAbs largely overlaps with that of gluten-specific T cells. Moreover, mAbs detect several homologous peptides present in gluten proteins. All except the LMW-specific mAbs also detect storage proteins present in barley and rye, whereas the gamma-gliadin-specific mAbs also recognize oat proteins.


CONCLUSIONS
The mAbs raised against T cell stimulatory peptides in gliadins and glutenins allow a comprehensive screen for the presence of harmful gluten and gluten-like proteins and peptides in food products. They can thus be used to guarantee the safety of food for celiac disease patients.
Department of Blood Transfusion and Immunohematology, Leiden University Medical Center, Leiden, Netherlands.
Cristina
C
Mitea
Yvonne
Y
Kooy-Winkelaar
Peter
P
van Veelen
Arnoud
A
de Ru
Jan W
JW
Drijfhout
Frits
F
Koning
Liesbeth
L
Dekking
D000911:Antibodies, Monoclonal / Q000276:immunology; D001667:Binding, Competitive; D015153:Blotting, Western; D002446:Celiac Disease / Q000276:immunology*; D003257:Consumer Product Safety; D004591:Electrophoresis, Polyacrylamide Gel; D005504:Food Analysis / Q000379:methods*; D005903:Gliadin / Q000276:immunology; D005983:Glutens / Q000276:immunology*; D006801:Humans; D008213:Lymphocyte Activation; D008970:Molecular Weight; D010446:Peptide Fragments / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*
D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D005983:Glutens; D005903:Gliadin; C037302:glutenin