Simultaneous flow cytometric detection of basophil activation marker CD63 and intracellular phosphorylated p38 mitogen-activated protein kinase in birch pollen allergy

Cytometry. Part B, Clinical Cytometry
Nicolaas E AertsDidier G Ebo

Abstract

Phosphorylation of p38 MAPK is a crucial step in IgE-receptor signaling in basophils. The relation of p38 MAPK to the well-validated diagnostic cell surface marker CD63 has not been evaluated in a clinical allergy model. Expression of CD63 and phosphorylation of p38 MAPK were analyzed flow cytometrically in anti-IgE-gated basophils from 18 birch pollen allergic patients, five grass pollen allergic patients, and five healthy individuals, after 3 and 20 min of stimulation with recombinant major birch pollen allergen (rBet v 1). Additional time points and the influence of p38 MAPK inhibitor SB203580 were studied in birch pollen allergic patients. Phospho-p38 MAPK and CD63 were expressed dose-dependently in birch pollen allergic patient basophils within 1 minute of rBet v 1 stimulation. P38 MAPK phosphorylation was fastest and subsided gradually while CD63 expression remained elevated for at least 20 min. Inhibition of p38 MAPK significantly inhibited CD63 upregulation. With optimal stimulation of the cells (1 μg/mL), sensitivity and specificity for the discrimination between patients and a group of control individuals (grass pollen allergic patients and healthy controls) were 94% and 100% for CD63 at 3 and 20 min and for phospho-p...Continue Reading

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Citations

Oct 27, 2010·International Archives of Allergy and Immunology·Yael GernezKari C Nadeau
Jul 16, 2014·Immunology and Allergy Clinics of North America·Astrid P UyttebroekDidier G Ebo
Aug 21, 2013·The Journal of Allergy and Clinical Immunology·Donald W MacGlashan
Dec 17, 2011·Cytometry. Part B, Clinical Cytometry·Vito SabatoDidier G Ebo
Nov 22, 2011·Cytometry. Part B, Clinical Cytometry·Marjoke M VerweijDidier G Ebo
Jan 11, 2011·Allergy, Asthma & Immunology Research·Salvatore ChirumboloAntonio Vella
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Jan 16, 2010·International Journal of Immunopathology and Pharmacology·B F Gibbs
Sep 17, 2014·Expert Review of Clinical Immunology·Astrid P UyttebroekDidier G Ebo
Jan 26, 2021·Journal of Immunological Methods·Anne-Emmanuelle BergerClaude Lambert

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