OBJECTIVE AND DESIGN: 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. MATERIALS AND SUBJECTS: Subjects allergic to grass pollen were selected by their clinical history, skin tests and specific IgE. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
OBJECTIVE AND DESIGN: 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. MATERIALS AND SUBJECTS: Subjects allergic to grass pollen were selected by their clinical history, skin tests and specific IgE. METHODS: 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. RESULTS: 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-CD41FITC 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. CONCLUSION: 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.
Authors: Didier G Ebo; Steven E Haine; Margo M Hagendorens; Chris H Bridts; Viviane M Conraads; Ann Vorlat; Chris J Vrints; Luc S De Clerck; Wim J Stevens Journal: Clin Drug Investig Date: 2004 Impact factor: 2.859
Authors: Shabnam Foroughi; Barbara Foster; Nayoung Kim; Leigh B Bernardino; Linda M Scott; Robert G Hamilton; Dean D Metcalfe; Peter J Mannon; Calman Prussin Journal: J Allergy Clin Immunol Date: 2007-09 Impact factor: 10.793
Authors: Theodore Kim; Jing Yu; Henry Li; Mark Scarupa; Richard L Wasserman; Athena Economides; Martha White; Carla Ward; Atul Shah; Douglas Jones; Melinda Rathkopf; Kelly Frye; Ahmet Aybar; Shahrooz Shayegan; Benjamin Enav; Laura Ispas; Denise Loizou; David Fitzhugh; James Tracy; James Friedlander; Zachary Jacobs; Jonathan Matz; David Golden; Donald McNeil; William McCann; Christopher Copenhaver; Jeffrey Factor; Raavi Gupta; Oral Alpan; Matthew Plassmeyer; Søren Ulrik Sønder Journal: Cytometry B Clin Cytom Date: 2021-02-04 Impact factor: 3.248
Authors: Kaori Mukai; Nicolas Gaudenzio; Sheena Gupta; Nora Vivanco; Sean C Bendall; Holden T Maecker; Rebecca S Chinthrajah; Mindy Tsai; Kari C Nadeau; Stephen J Galli Journal: J Allergy Clin Immunol Date: 2016-07-15 Impact factor: 10.793