1. Optimizing of the basophil activation test: Comparison of different basophil identification markers.
- Author
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Eberlein B, Hann R, Eyerich S, Pennino D, Ring J, Schmidt-Weber CB, and Buters J
- Subjects
- Adult, Allergens chemistry, Allergens immunology, Antibodies, Monoclonal pharmacology, Basophil Degranulation Test, Basophils drug effects, Basophils pathology, CD3 Complex genetics, CD3 Complex immunology, Case-Control Studies, Cells, Cultured, Female, Flow Cytometry methods, Gene Expression, Humans, Male, Middle Aged, N-Formylmethionine Leucyl-Phenylalanine pharmacology, Phosphoric Diester Hydrolases genetics, Phosphoric Diester Hydrolases immunology, Plant Extracts chemistry, Plant Extracts immunology, Plant Extracts pharmacology, Pollen chemistry, Pollen immunology, Pyrophosphatases genetics, Pyrophosphatases immunology, Receptors, CCR3 genetics, Receptors, CCR3 immunology, Receptors, Immunologic genetics, Receptors, Immunologic immunology, Receptors, Prostaglandin genetics, Receptors, Prostaglandin immunology, Respiratory Hypersensitivity immunology, Respiratory Hypersensitivity pathology, Tetraspanin 30 genetics, Tetraspanin 30 immunology, Antibodies, Anti-Idiotypic chemistry, Basophils immunology, Immunoglobulin E blood, Immunophenotyping methods, Respiratory Hypersensitivity diagnosis
- Abstract
Background: Flowcytometric identification of basophils is a prerequisite for measuring activation of basophils with IgE-dependent or IgE-independent stimuli. Aim of this study was to compare different marker combinations in a simultaneous multicolor flowcytometric measurement., Methods: Ten patients with a grass pollen allergy and three controls were included in the study. Basophilic cells were gated by using anti-CCR3, anti-IgE, anti-CRTH2, anti-CD203c, and anti-CD3. Cells were activated by a monoclonal anti-FcεRI antibody, N-formyl-methionyl-leucyl-phenylalanine (fMLP), and the allergen extract Phleum pratense. The activation marker anti-CD63 was used., Results: The highest relative number of basophils was found with anti-CCR3+ cells, anti-IgE+ and anti-IgE+ /anti-CD203c+ cells, the lowest with CRTH2+/CD203c+/CD3- cells. A very good and good concordance of CCR3+ cells was seen with CCR3+/CD3- cells and CRTH2+/CD203c+/CD3- cells in all experiments. The contamination of the CCR3+ population with CD3+ cells and the contamination of the IgE+-population with CCR3- cells and CD203- cells were the lowest compared to all other marker combinations., Conclusions: As the highest relative number of basophils was identified by anti-CCR3 followed by the anti-IgE and anti-IgE/antiCD203c positive population in most cases, these markers can generally be recommended for identification of basophils. If a basophil population with very high purity is needed, anti-IgE should be chosen., (© 2014 International Clinical Cytometry Society.)
- Published
- 2015
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