Extracts of wheat gluten activate complement via the alternative pathway.
We studied the ability of wheat gluten and its subfractions to activate complement directly. A sensitive sandwich ELISA employing a monoclonal antibody (MoAb) to a C9 neoepitope exposed in the terminal complement complex (TCC), a functional haemolytic assay for C5b6 generation, and Laurell's electrophoretic method of estimating C3 conversion to C3bi were used. On a weight-for-weight basis, enzyme solubilized Frazer's fraction three of gluten (FIII) produced approximately 75% of the complement activation seen with the potent activator zymosan. By contrast, activation with whole insoluble undigested gluten was very weak and similar to that seen with ovalbumin or beta-lactoglobulin. The results were the same using normal human serum or sera from patients with coeliac disease, dermatitis herpetiformis, or hypogammaglobulinaemia as the complement source. Activation by both zymosan and FIII was blocked in 0.01 M EDTA, but not in 0.01 M EGTA with 0.0025 M magnesium chloride. Zymosan and FIII activated complement in a serum from a patient with an intact alternative pathway but classical pathway haemolytic activity (CH50) of zero. Preferential heat inactivation of the alternative pathway inhibited both zymosan- and FIII-induced activation. Our results confirm that FIII is a strong activator of the alternative pathway. We discuss how gluten enteropathy might be initiated by complement.
Unsworth
D J
DJ
Department of Clinical Immunology, Addenbrooke's Hospital, Cambridge, UK.
Würzner
R
R
Brown
D L
DL
Lachmann
P J
PJ
D000328:Adult; D000361:Agammaglobulinemia / Q000276:immunology; D000911:Antibodies, Monoclonal; D002446:Celiac Disease / Q000276:immunology; D003167:Complement Activation / Q000276:immunology*; D015941:Complement Hemolytic Activity Assay; D003170:Complement Pathway, Alternative / Q000276:immunology*; D003874:Dermatitis Herpetiformis / Q000276:immunology; D004492:Edetic Acid; D004533:Egtazic Acid; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005983:Glutens / Q000276:immunology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D010936:Plant Extracts / Q000276:immunology; D014908:Triticum / Q000276:immunology*; D015054:Zymosan / Q000276:immunology
D000911:Antibodies, Monoclonal; D010936:Plant Extracts; D004533:Egtazic Acid; D005983:Glutens; D015054:Zymosan; D004492:Edetic Acid