[Application of flow cytometry to the analysis of activation of human basophils. Immunologic validation of the method].
Among the different methods used for the analysis of human basophil activation as histamine and leukotriene release, flow cytometry presents several methodological advantages. Moreover, preliminary results obtained for a wide allergen panel have demonstrated its high sensitivity and specificity. We present here the results of the methods used to validate the flow cytometric analysis of human basophil activation by double anti-IgE FITC/anti-CD63 PE labelling of human leucocytes. The use of multiple labelling by monoclonals having different specificities have shown that the selected cells express the phenotype CD14-, CD19-, CD41- or +/-, CD45+, CD63- or +. This phenotype is related to human basophils. We have also demonstrated that the expression of CD63 is related to basophil activation and not to aggregated platelets which are also able to express this marker.
Sainte-Laudy
J
J
Unité d'immuno-allergologie, Lavoratoire Cerba, Cergy-Pontoise.
D015703:Antigens, CD / Q000032:analysis; D018941:Antigens, CD19 / Q000032:analysis; D001491:Basophils / Q000166:cytology / Q000276:immunology*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis; D007132:Immunoglobulin Isotypes / Q000276:immunology; D017493:Leukocyte Common Antigens / Q000032:analysis; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D008213:Lymphocyte Activation; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000032:analysis; D010980:Platelet Membrane Glycoproteins / Q000032:analysis; D060149:Tetraspanin 30
D015703:Antigens, CD; D018941:Antigens, CD19; C554586:CD63 protein, human; D007132:Immunoglobulin Isotypes; D018950:Lipopolysaccharide Receptors; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D010980:Platelet Membrane Glycoproteins; D060149:Tetraspanin 30; D007073:Immunoglobulin E; D017493:Leukocyte Common Antigens