Optimizing of the basophil activation test: Comparison of different basophil identification markers
| Autor: | Eberlein B, Hann R, Eyerich S, Pennino D, Ring J, Cb, Schmidt-Weber, Jeroen Buters |
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| Rok vydání: | 2014 |
| Předmět: |
Adult
Male Basophil Activation Test Basophil Identification Markers Multicolor Flowcytometry CD3 Complex Receptors CCR3 Receptors Prostaglandin Basophil Degranulation Test Gene Expression Immunophenotyping Respiratory Hypersensitivity Humans Pyrophosphatases Receptors Immunologic Cells Cultured Phosphoric Diester Hydrolases Plant Extracts Tetraspanin 30 Antibodies Monoclonal hemic and immune systems Allergens Immunoglobulin E Middle Aged Flow Cytometry Antibodies Anti-Idiotypic Basophils N-Formylmethionine Leucyl-Phenylalanine Case-Control Studies Pollen Female |
| Zdroj: | Cytometry B Clin. Cytom. 88, 183-189 (2015) Europe PubMed Central |
| ISSN: | 1552-4957 |
| Popis: | 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 3 controls were included in the study. Basophilic cells were gated by using anti-CCR3647, anti-IgE, anti-CRTH2, anti-CD203c und 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-CD193 (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. |
| Databáze: | OpenAIRE |
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