In vitro assays for the diagnosis of IgE-mediated disorders

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Presentation transcript:

In vitro assays for the diagnosis of IgE-mediated disorders Robert G. Hamilton, PhD, N. Franklin Adkinson, MD Journal of Allergy and Clinical Immunology Volume 114, Issue 2, Pages 213-225 (August 2004) DOI: 10.1016/j.jaci.2004.06.046 Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 1 Schematic diagram of the original RAST for IgE antibody developed in 1967. In the first incubation allergen-specific antibody is extracted from serum with an allergosorbent. After buffer washes to remove unbound serum proteins, bound IgE antibody is detected with an iodine 125–labeled rabbit anti-human IgE Fc detection antibody. The counts per minute (CPM) bound to the allergosorbent are proportional to the amount of specific IgE in the original serum. Even in this earliest assay, quantitative IgE antibody results were emphasized. The test serum IgE antibody results were obtained at multiple serum dilutions, interpolated from a multipoint dose-response curve, corrected for dilution, and reported in units relative to the quantity of IgE antibody in the original serum. Reproduced with permission from Adkinson.24 Journal of Allergy and Clinical Immunology 2004 114, 213-225DOI: (10.1016/j.jaci.2004.06.046) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 2 Allergen microarray. A, Scheme of environmental allergens spotted in triplicate in vertical columns. B and C, Scanned images from a microarray chip–based IgE assay obtained with serum from an allergic (Fig 2, B) and nonallergic (Fig 2, C) individual. D, Dot-blot IgE reactivity to the same allergens bound to nitrocellulose. E, Skin reactivity to allergens and histamine (H6; wheal circled for visualization). Reproduced with permission from Hiller et al.30 Journal of Allergy and Clinical Immunology 2004 114, 213-225DOI: (10.1016/j.jaci.2004.06.046) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 3 Probability curves from a retrospective and prospective study of patients reacting to 4 foods (egg white, cow's milk, fish, and peanut) given a measured quantitative level of allergen-specific IgE antibody. The food allergen–specific IgE 95% decision level (kUa/L) above which a double-blind, placebo-controlled food challenge is not considered necessary is as follows: egg, 7 kUa/L; milk, 15 kUa/L; peanut, 14 kUa/L; and fish, 10 kUa/L. Not shown in the figure are the 95% decision levels for soy (65 kUa/L) and wheat (80 kUa/L). Reproduced with permission from Sampson.15 Journal of Allergy and Clinical Immunology 2004 114, 213-225DOI: (10.1016/j.jaci.2004.06.046) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 4 Selected curves demonstrating the probability of receiving a positive allergy diagnosis at a given IgE antibody level for different allergens at 3 different clinics. Each curve's shape demonstrates the allergist's disposition for a positive clinical allergy diagnosis in relation to the level of allergen-specific IgE antibody in serum. The different slopes indicate different identification patterns of symptoms. A flat slope reflects difficulty in linking high IgE antibody levels as a trigger for allergic symptoms. A steep slope indicates that allergic symptoms are easily linked to low levels of allergen-specific IgE antibody. Extracted and reproduced with permission from Soderstrom et al.16 Journal of Allergy and Clinical Immunology 2004 114, 213-225DOI: (10.1016/j.jaci.2004.06.046) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions