In vitro Analysis of Birch-Pollen-Associated Food Allergy by Use of Recombinant Allergens in the Basophil Activation Test

• Published in: International Archives of Allergy and Immunology Vol. 136; no. 3; pp. 230 - 238
• Main Authors: Erdmann, S.M., Sachs, B., Schmidt, A., Merk, H.F., Scheiner, O., Moll-Slodowy, S., Sauer, I., Kwiecien, R., Maderegger, B., Hoffmann-Sommergruber, K.
• Format: Journal Article
• Language: English
• Published: Basel, Switzerland Karger 01.03.2005; S. Karger AG
• Subjects: Allergens - analysis; Allergens - immunology; Allergies; Antibodies, Monoclonal; Antigens, CD - analysis; Antigens, CD - biosynthesis; Antigens, Plant; Apium graveolens; Basophils - immunology; Betula - immunology; Betula verrucosa; Biological and medical sciences; Daucus carota; Diagnostic tests; Europe - epidemiology; Flow Cytometry - methods; Food; Food Hypersensitivity - blood; Food Hypersensitivity - diagnosis; Food Hypersensitivity - epidemiology; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Humans; Immunoglobulin E - analysis; Immunoglobulin E - blood; Immunopathology; In Vitro Techniques; Malus dometicus; Medical sciences; Original Paper; Plant Proteins - immunology; Platelet Membrane Glycoproteins - analysis; Platelet Membrane Glycoproteins - biosynthesis; Pollen; Pollen - immunology; Recombinant Proteins - immunology; Sensitivity and Specificity; Skin Tests; Tetraspanin 30; Flowcytometry; Food allergy; Dau c 1; CD63 expression; Recombinant allergens; Api g 1; Mal d 1; Basophil activation test; Bet v 1; Birch; Insecta; Granulocyte; Apidae; Basophil; Pollen analysis; Immunology; Apoidea; Recombinant protein; Aculeata; Immunopathology; Apis; In vitro; Arthropoda; Hymenoptera; Invertebrata
• ISSN: 1018-2438; 1423-0097; 1365-2567
• DOI: 10.1159/000083949

Background: Basophil activation is associated with the expression of CD63. In birch-pollen-associated food allergy to celery, carrot and apple, Bet v 1, Api g 1, Dau c 1 and Mal d 1 are major allergens. Recombinant allergens have not yet been used in the CD63-based basophil activation test (BAT). Objective: To evaluate the feasibility of using recombinant allergens in the BAT in the diagnosis of allergy to apple, carrot and celery and to compare results with routine tests, i.e. skin prick tests (SPTs) and specific IgE. Methods: Thirty-two patients with an oral allergy syndrome induced by apple, carrot or celery and 22 controls were studied. SPTs were performed with native foods. Specific IgE was determined by the CAP method and basophil activation by flowcytometry upon double staining with anti-IgE/anti-CD63 monoclonal antibodies after incubating with purified recombinant Bet v 1, Bet v 2, Api g 1, Dau c 1 and Mal d 1. Results: By the combined use of the BAT and the CAP method, sensitization to Bet v 1 and Bet v 2 was detected in 100 and 25% of all subjects, respectively. Sensitivity of specific IgE for apple, carrot and celery was 60, 70 and 75% with corresponding specificities of 64, 86 and 82%. Sensitivity of the BAT for Mal d 1, Dau c 1 and Api g 1 was 75, 65 and 75% with corresponding specificities of 68, 100 and 77%. Conclusions: The BAT using recombinant allergens provides a valuable new in vitro method for the detection of sensitization to foods. Although double-blind placebo-controlled food challenges remain the gold standard to confirm food allergy, the CD63-based BAT with recombinant allergens may supplement routine tests for allergy diagnosis.