Flow cytometric allergy diagnosis : basophil activation techniques
Faculty of Medicine and Health Sciences
New York, N.Y. :Springer, 2014
Basophils and mast cells : methods and protocols / Gibbs, Bernard F. [edit.]; et al.
Methods in molecular biology ; 1192
University of Antwerp
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: 332339, 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: 201210, 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.