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Allergy pp 33–43Cite as

Measurement of Allergen-Specific Inhibitory Antibody Activity

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2020))

Abstract

Specific allergen immunotherapy (AIT) is an effective treatment for IgE-mediated allergic diseases and involves T- and B-cell-mediated events. IgE receptors on the surface of antigen-presenting cells facilitate the presentation of allergens in the presence of specific IgE antibody resulting in T-cell activation. Interference with these IgE-dependent mechanisms by “blocking” IgG antibodies suppresses pro-inflammatory Th2 cell responses and manifests as a reduction in allergic responses in vivo.

In vitro assays used to measure the inhibition of binding of allergen-IgE complexes have previously utilized proliferation of antigen-specific T-cell clones as an assay readout. Here we describe two simplified assays to measure allergen binding without the complexity of generating T-cell clones. The IgE-facilitated allergen binding assay (IgE-FAB) utilizes flow cytometry to measure the binding of allergen-IgE complexes to EBV-transformed B cells. The enzyme-linked immunosorbent-facilitated antigen binding (ELIFAB) assay uses standard ELISA-based techniques to measure allergen-IgE binding to plate-bound CD23, the low-affinity IgE receptor expressed on B cells.

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Acknowledgments

The authors would like to thank Peter Würtzen, Stephen J Till, Rebecca Parkin, and Natalia Do Couto Francisco for their contributions to this protocol.

Author information

Authors and Affiliations

  1. Immunomodulation and Tolerance Group, Allergy and Clinical Immunology, Inflammation, Repair and Development, National Heart and Lung Institute, Imperial College London, London, UK

    Mohamed H. Shamji

  2. Asthma UK Centre in Allergic Mechanisms of Asthma, London, UK

    Mohamed H. Shamji

  3. Immune Targeting Systems (ITS) Ltd., London, UK

    James N. Francis

Authors
  1. Mohamed H. Shamji
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  2. James N. Francis
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Corresponding author

Correspondence to Mohamed H. Shamji .

Editor information

Editors and Affiliations

  1. Core Facilities, St. George’s University of London, London, UK

    Penny Lympany

  2. Department of Occupational and Environmental Medicine, Imperial College London, London, UK

    Meinir G. Jones

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Shamji, M.H., Francis, J.N. (2019). Measurement of Allergen-Specific Inhibitory Antibody Activity. In: Lympany, P., Jones, M. (eds) Allergy. Methods in Molecular Biology, vol 2020. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9591-2_3

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  • DOI: https://doi.org/10.1007/978-1-4939-9591-2_3

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9589-9

  • Online ISBN: 978-1-4939-9591-2

  • eBook Packages: Springer Protocols

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