Analysis of anti-IgE and allergen induced human basophil activation by flow cytometry. Comparison with histamine release

On the basis of flow cytometric methods previously described for the analysis of human basophil activation, we present here a bi-color anti-IgE FITC, anti CD63 PE method and the correlation with histamine release. Subjects allergic to grass pollen were selected by their clinical history, skin tests...

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Bibliographic Details
Published in:Inflammation research 1998-10, Vol.47 (10), p.401-408
Main Authors: Sainte-Laudy, J, Sabbah, A, Vallon, C, Guerin, J C
Format: Article
Language:English
Subjects:
Allergens - immunology
Antibodies, Anti-Idiotypic
Antigens, CD - analysis
Basophils - immunology
Basophils - physiology
Blood Platelets - immunology
Flow Cytometry
Fluorescein-5-isothiocyanate
Fluorescent Dyes
Histamine Release
Humans
Immunoglobulin E - analysis
Immunoglobulin E - immunology
Immunophenotyping
Platelet Glycoprotein GPIIb-IIIa Complex - analysis
Platelet Membrane Glycoproteins - analysis
Sensitivity and Specificity
Tetraspanin 30
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Summary:On the basis of flow cytometric methods previously described for the analysis of human basophil activation, we present here a bi-color anti-IgE FITC, anti CD63 PE method and the correlation with histamine release. Subjects allergic to grass pollen were selected by their clinical history, skin tests and specific IgE. Basophils gated in the lymphocyte region of the side scatter (SSC)/forward scatter (FSC) pattern were selected by their high IgE epitope density. Percentage of cells expressing CD63 marker, upregulated on activated basophil membrane, was calculated by the cytometer. Histamine released into the supernatants was measured by RIA. In these conditions, flow cytometric analysis of blood leukocytes showed that the selected cells had the phenotype CD14-, CD19-, CD45+, IgE++ and CD63- or + which is related to human basophil phenotype, the isotype controls being negative. The use of an anti-CD41 FITC antibody also showed the presence of aggregated platelets on the basophil membrane, CD63 antigen being, however, expressed by basophils themselves and not by platelets. Moreover, no statistical difference was observed between histamine release and flow cytometry after passive sensitization of blood donor leukocytes. Flow cytometry, as a popular method often used in the immunology and haematology departments of clinical laboratories may represent a new alternative for allergy diagnosis and basophil pharmacology.
ISSN:1023-3830
1420-908X
DOI:10.1007/s000110050351