Measurement of IgE, IgG1 and IgG4 antibodies against mite Sephadex fractions and purified allergens by means of enzyme-linked immunosorbent assay

Mite antigens (Dermatophagoides farinae) were fractionated by a Sephadex G-200 column and their reactivities with IgE, IgG1 and IgG4 antibodies were investigated with enzyme-linked immunosorbent assay (ELISA). High IgE antibody values were observed in fractions with low molecular weight (allergenic part), while high IgG1 and IgG4 antibody values were observed in fractions with high molecular weight. High IgG4 antibody values to crude mite extract and fractions with high molecular weight were detected in individuals who had received immunotherapy. However, IgG4 antibodies directed to allergenic part were found in only one out of 12 sera tested. IgG4-ELISA using DF1 (major allergen of Dermatophagoides farinae) as antigen was also performed. In the group treated with mite, significant IgG4 antibody levels were detected in only one out of 13 sera tested. In the group treated with house dust, significant IgG4 antibodies were detected in only one out of 12 sera tested. Patients who showed high IgG4 antibody responses to crude mite extract and to high molecular weight did not show responses to allergenic part and DF1. The only case who showed positive IgG4 responses to allergenic part also reacted with DF1. Those results suggest that IgG1 and IgG4 antibody values in ELISA using crude mite extract as antigen do not reflect major allergen-specific antibody values. The importance of the use of partially purified antigens in measuring major allergen-specific IgG4 antibodies was also suggested.