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Correlation of IgE/IgG4 milk epitopes and affinity of milk-specific IgE antibodies with different phenotypes of clinical milk allergy  Julie Wang, MD,

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Presentation on theme: "Correlation of IgE/IgG4 milk epitopes and affinity of milk-specific IgE antibodies with different phenotypes of clinical milk allergy  Julie Wang, MD,"— Presentation transcript:

1 Correlation of IgE/IgG4 milk epitopes and affinity of milk-specific IgE antibodies with different phenotypes of clinical milk allergy  Julie Wang, MD, Jing Lin, PhD, Ludmilla Bardina, MSc, Marina Goldis, MD, Anna Nowak-Węgrzyn, MD, Wayne G. Shreffler, MD, PhD, Hugh A. Sampson, MD  Journal of Allergy and Clinical Immunology  Volume 125, Issue 3, Pages e6 (March 2010) DOI: /j.jaci Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Comparison of IgE and IgG4 binding frequency to peptides of 5 milk allergens between the different clinical phenotypes. The x-axis shows the overlapping peptides, and the y-axis shows the percentage of patients within each group showing positive binding to each peptide. Binding for IgE is indicated above the x-axis line; binding for IgG4 is below the line. Candidates of informative epitopes are indicated with numbers. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Heat map of IgE binding to candidates of informative epitopes from the allergic subjects compared with the HM-tolerant/outgrown subjects. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Amino acid sequence of milk allergens. Candidates of informative epitopes identified based on the binding frequency are in bold and underlined. Informative epitopes recognized by greater than 50% of the allergic patients with high-affinity IgE antibodies are highlighted in gray. The numbers and letters indicated on top of each epitope correspond to the ones indicated in the other figures. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Heat map showing the effects of competition on 7 allergic and 7 HM-tolerant/outgrown subjects. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Scatter plot comparing IgE binding from 7 allergic (red) and 7 HM-tolerant/outgrown (blue) subjects between the standard protocol (x-axis) and the competition assay (y-axis). Each spot represents IgE binding (represented as a z score) to 1 milk peptide. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 Comparison of IgE peptide-binding frequency between groups using the standard protocol and competition assay. Informative epitopes recognized by greater than 50% of the allergic patients with high-affinity IgE antibodies are indicated with letters. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Scatter plot showing the correlation of IgE and IgG4 binding of 8 randomly selected subjects from each group. Each spot represents IgE binding (represented as a z score) to 1 milk peptide. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Distribution of IgE-binding peptides for each group.
Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Effect of temperature. Scatter plot comparing IgE binding between the standard protocol (x-axis) and competition with 2 mg/mL caseins and 3 mg/mL β-lactoglobulin at different temperatures (y-axis) for 4 hours. Each spot represents IgE binding (represented as a z score) to 1 peptide of milk allergens. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

11 Effect of competitor concentration
Effect of competitor concentration. Scatter plot comparing IgE binding of the serum pool between the standard protocol (x-axis) and competition with a one-third serial dilution of 2 mg/mL caseins and 3 mg/mL β-lactoglobulin (y-axis) at 16°C for 4 hours. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

12 Effect of competition time
Effect of competition time. Scatter plot comparing IgE binding of a subject with milk allergy and a subject who had outgrown his or her allergy between the standard protocol (x-axis) and 1 mg/mL caseins and 1.5 mg/mL β-lactoglobulin (y-axis) at 16°C for different competition time. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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