title: Flow cytometric allergy diagnosis: basophil activation techniques.
abstract: The basis of flow cytometric allergy diagnosis is quantification of changes in 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 up-regulate 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 detection of surface marker activation (Method 1) and changes in intragranular histamine (Method 2), both reflecting different facets of basophil activation.
authors.affiliation: Department of Immunology and Allergology, University of Antwerp, Campus Drie Eiken, D.T.596, Universiteitsplein 1, 2610, Wilrijk, Belgium.
authors.forename: Chris H
authors.initials: CH
authors.lastname: Bridts
authors.forename: Vito
authors.initials: V
authors.lastname: Sabato
authors.forename: Christel
authors.initials: C
authors.lastname: Mertens
authors.forename: Margo M
authors.initials: MM
authors.lastname: Hagendorens
authors.forename: Luc S
authors.initials: LS
authors.lastname: De Clerck
authors.forename: Didier G
authors.initials: DG
authors.lastname: Ebo
mesh_terms: D015549:Basophil Degranulation Test / Q000379:methods*; D001491:Basophils / Q000276:immunology* / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002138:Calibration; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards; D006632:Histamine / Q000032:analysis / Q000378:metabolism; D006801:Humans; D006967:Hypersensitivity / Q000175:diagnosis* / Q000276:immunology; D013194:Staining and Labeling / Q000379:methods
chemical_list: D015415:Biomarkers; D006632:Histamine