Measurement of murine IgE antibody responses to dust mite allergens by in vitro assay.
In an analysis of murine immune responses to the dust mite allergen Der p 1, treatment with purified allergen induced a significant increase in the level of circulating IgE immunoglobulin (from less than 100 ng/ml in normal mice to 1,350 ng/ml in mice receiving the allergen). Even so, specific IgE antibodies binding to purified Der p 1 were not detected in a conventional ELISA, and the major response appeared to be the induction of high titre IgG antibodies. Specific circulating murine IgE antibodies were however detected using the following assay format: murine IgE was captured to anti-murine IgE antibody coated wells; Der p 1 was added and bound by immobilized anti-Der p 1 IgE antibodies; the captured Der p 1 was then detected by the addition of monoclonal IgG antibodies against Der p 1 and these antibodies were measured by the addition of anti-murine IgG antibody-enzyme conjugate with which colour development is produced after substrate addition. This assay establishes a procedure to measure circulating anti-Der p 1 IgE antibodies which are present together with competing high titre IgG anti-Der p 1 antibodies.
Ma
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Cancer Research Laboratory, School of Pharmaceutical Sciences, University of Nottingham, Nottingham NG7 2RD, UK.
Gunn
B
B
Byers
V S
VS
Baldwin
R W
RW
Price
M R
MR
D000485:Allergens / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000941:Antigens / Q000276:immunology; D039741:Antigens, Dermatophagoides; D001667:Binding, Competitive / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006023:Glycoproteins / Q000276:immunology*; D007073:Immunoglobulin E / Q000097:blood*; D007074:Immunoglobulin G / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D008925:Mites / Q000276:immunology
D000485:Allergens; D000911:Antibodies, Monoclonal; D000941:Antigens; D039741:Antigens, Dermatophagoides; D006023:Glycoproteins; D007074:Immunoglobulin G; D007073:Immunoglobulin E