Determination of IgG subclass antibodies against wheat flour antigens by an ELISA technique.
We describe the assay conditions for an enzyme-linked immunoassay for the determination of IgG and IgG subclass antibodies in serum to water-soluble wheat flour antigens. The optimal antigen coating concentration was 5 micrograms/ml for total IgG, IgG1, IgG4 and 100 micrograms/ml for IgG2. Serial dilutions of test sera were used and commercially available monoclonal mouse anti-human IgG isotype antibodies (as ascites fluid) were diluted 1:500-1:1000. Specific wheat flour antibodies belonging to the IgG1, IgG2 and IgG4 subclasses were detected. Despite the lack of standardized isotype-specific second mouse monoclonal antibodies, the subclass antibody levels between flour-exposed bakers and controls could be compared. We observed significantly higher IgG1, IgG2, and IgG4 subclass antibodies among 23 bakers than among 12 non-exposed controls, but no IgG3 antibodies were detected. The differences in biological activities of the IgG subclass antibodies may explain the clinical and pathophysiological features for fluor-induced occupational allergic diseases among bakers.
Tiikkainen
U
U
Institute of Occupational Health, Helsinki, Finland.
Klockars
M
M
D000293:Adolescent; D000328:Adult; D000918:Antibody Specificity; D001249:Asthma / Q000276:immunology*; D004391:Dust / Q000009:adverse effects; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005260:Female; D005433:Flour / Q000009:adverse effects*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis*; D007132:Immunoglobulin Isotypes / Q000032:analysis*; D008297:Male; D008875:Middle Aged; D009784:Occupational Diseases / Q000276:immunology*; D012130:Respiratory Hypersensitivity / Q000276:immunology*; D014908:Triticum / Q000276:immunology*
D004391:Dust; D000939:Epitopes; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes