Abstract
The basis of flow cytometric allergy diagnosis is the quantification of changes in the expression of basophilic surface membrane markers (Ebo et al., Clin Exp Allergy 34: 332–339, 2004). Upon encountering specific allergens recognized by surface receptor FcεRI-bound IgE, basophils not only secrete and generate quantifiable bioactive mediators but also upregulate the expression of different markers (e.g., CD63, CD203c) which can be detected by multicolor flow cytometry using specific monoclonal antibodies (Ebo et al., Cytometry B Clin Cytom 74: 201–210, 2008). Here, we describe two flow cytometry-based protocols which allow the detection of surface marker activation (Method 1) and changes in intragranular histamine (Method 2), both reflecting different facets of basophil activation.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Ebo DG, Hagendorens MM, Bridts CH, Schuerwegh AJ, De Clerck LS, Stevens WJ (2004) In vitro allergy diagnosis: should we follow the flow? Clin Exp Allergy 34:332–339
Ebo DG, Bridts CH, Hagendorens MM, Aerts NE, De Clerck LS, Stevens WJ (2008) Basophil activation test by flow cytometry: present and future applications in allergology. Cytometry B Clin Cytom 74:201–210
Hausmann OV, Gentinetta T, Bridts CH, Ebo DG (2009) The basophil activation test in immediate-type drug allergy. Immunol Allergy Clin N Am 29:555–566
Ebo DG, Bridts CH, Hagendorens MM, Mertens CH, De Clerck LS, Stevens WJ (2006) Flow-assisted diagnostic management of anaphylaxis from rocuronium bromide. Allergy 61:935–939
Sabato V, Verweij MM, Bridts CH, Levi-Schaffer F, Gibbs BF, De Clerck LS et al (2012) CD300a is expressed on human basophils and seems to inhibit IgE/FcepsilonRI-dependent anaphylactic degranulation. Cytometry B Clin Cytom 82:132–138
Ebo DG, Dombrecht EJ, Bridts CH, Aerts NE, De Clerck LS, Stevens WJ (2007) Combined analysis of intracellular signalling and immunophenotype of human peripheral blood basophils by flow cytometry: a proof of concept. Clin Exp Allergy 37:1668–1675
Verweij MM, De Knop KJ, Bridts CH, De Clerck LS, Stevens WJ, Ebo DG (2010) P38 mitogen-activated protein kinase signal transduction in the diagnosis and follow up of immunotherapy of wasp venom allergy. Cytometry B Clin Cytom 78:302–307
Verweij MM, Sabato V, Nullens S, Bridts CH, De Clerck LS, Stevens WJ et al (2012) STAT5 in human basophils: IL-3 is required for its FcepsilonRI-mediated phosphorylation. Cytometry B Clin Cytom 82:101–106
Ebo DG, Bridts CH, Mertens CH, Hagendorens MM, Stevens WJ, De Clerck LS (2012) Analyzing histamine release by flow cytometry (HistaFlow): a novel instrument to study the degranulation patterns of basophils. J Immunol Methods 375:30–38
Nullens S, Sabato V, Faber M, Leysen J, Bridts CH, De Clerck LS et al (2013) Basophilic histamine content and release during venom immunotherapy: insights by flow cytometry. Cytometry B Clin Cytom 84:173–178
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Bridts, C.H., Sabato, V., Mertens, C., Hagendorens, M.M., De Clerck, L.S., Ebo, D.G. (2020). Flow Cytometric Allergy Diagnosis: Basophil Activation Techniques. In: Gibbs, B.F., Falcone, F.H. (eds) Basophils and Mast Cells. Methods in Molecular Biology, vol 2163. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0696-4_15
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0696-4_15
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0695-7
Online ISBN: 978-1-0716-0696-4
eBook Packages: Springer Protocols