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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References
Durham SR, Walker SM, Varga EM et al (1999) Long-term clinical efficacy of grass-pollen immunotherapy. N Engl J Med 341:468–475
James LK, Shamji MH, Walker SM et al (2011) Long-term tolerance after allergen immunotherapy is accompanied by selective persistence of blocking antibodies. J Allergy Clin Immunol 127:509–516
Durham SR, Emminger W, Kapp A et al (2012) SQ-standardized sublingual grass immunotherapy: confirmation of disease modification 2 years after 3 years of treatment in a randomized trial. J Allergy Clin Immunol 129:717–725
Wambre E, DeLong JH, James EA et al (2012) Differentiation stage determines pathologic and protective allergen-specific CD4+ T-cell outcomes during specific immunotherapy. J Allergy Clin Immunol 129:544–551
Francis JN, Till SJ, Durham SR (2003) Induction of IL-10+CD4+CD25+ T cells by grass pollen immunotherapy. J Allergy Clin Immunol 111:255–261
O’Hehir RE, Gardner LM, de Leon MP et al (2009) House dust mite sublingual immunotherapy: the role for transforming growth factor-beta and functional regulatory T cells. Am J Respir Crit Care Med 180:936–947
Swamy RS, Reshamwala N, Hunter T et al (2012) Epigenetic modifications and improved regulatory T-cell function in subjects undergoing dual sublingual immunotherapy. J Allergy Clin Immunol 130:215–217
Nouri-Aria KT, Wachholz PA, Francis JN et al (2004) Grass pollen immunotherapy induces mucosal and peripheral IL-10 responses and blocking IgG activity. J Immunol 172:3252–3259
Shamji MH, Ljørring C, Francis JN et al (2012) Functional rather than immunoreactive levels of IgG4 correlate closely with clinical response to grass pollen immunotherapy. Allergy 67:217–226
van de Veen W, Stanic B, Yaman G et al (2013) IgG4 production is confined to human IL-10-producing regulatory B cells that suppress antigen-specific immune responses. J Allergy Clin Immunol 131:1204–1212
Garcia BE, Sanz ML, Gato JJ et al (1993) IgG4 blocking effect on the release of antigen-specific histamine. J Investig Allergol Clin Immunol 3:26–33
van der Heijden FL, van Neerven RJ, Kapsenberg ML (1995) Relationship between facilitated allergen presentation and the presence of allergen-specific IgE in serum of atopic patients. Clin Exp Immunol 99:289–293
van Neerven RJ, Wikborg T, Lund G et al (1999) Blocking antibodies induced by specific allergy vaccination prevent the activation of CD4+ T cells by inhibiting serum-IgE-facilitated allergen presentation. J Immunol 163:2944–2952
van Neerven RJ, Arvidsson M, Ipsen H et al (2004) A double-blind, placebo-controlled birch allergy vaccination study: inhibition of CD23-mediated serum immunoglobulin E-facilitated allergen presentation. Clin Exp Allergy 34:420–428
Wachholz PA, Soni NK, Till SJ, Durham SR (2003) Inhibition of allergen-IgE binding to B cells by IgG antibodies after grass pollen immunotherapy. J Allergy Clin Immunol 112:915–922
Shamji MH, Wilcock LK, Wachholz PA et al (2006) The IgE-facilitated allergen binding (FAB) assay: validation of a novel flow-cytometric based method for the detection of inhibitory antibody responses. J Immunol Methods 317:71–79
Kinaciyan T, Jahn-Schmid B, Radakovics A et al (2007) Successful sublingual immunotherapy with birch pollen has limited effects on concomitant food allergy to apple and the immune response to the Bet v 1 homolog Mal d 1. J Allergy Clin Immunol 119:937–943
Klunker S, Saggar LR, Seyfert V et al (2007) Combination treatment with omalizumab and rush immunotherapy for ragweed allergic rhinitis: induction of inhibitory antibody responses. J Allergy Clin Immunol 120:688–695
Jones SM, Pons L, Roberts JL, Scurlock AM et al (2009) Clinical efficacy and immune regulation with peanut oral immunotherapy. J Allergy Clin Immunol 24:292–300
Varga EM, Francis JN, Zach MS et al (2009) Time course of serum inhibitory activity for facilitated allergen-IgE binding (FAB) during bee venom immunotherapy in children. Clin Exp Allergy 39:1353–1357
Jones M, Jeal H, Schofield S et al (2014) Rat-specific IgG and IgG4 antibodies associated with inhibition of IgE-allergen complex binding in laboratory animal workers. Occup Environ Med 71:619–623
Shamji MH, Francis JN, Wurtzen PA et al (2013) Cell-free detection of allergen-IgE cross-linking with immobilized phase CD23: inhibition by blocking antibody responses after immunotherapy. J Allergy Clin Immunol 132:1003–1004
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.
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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
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