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Optimizing of the basophil activation test: Comparison of different basophil identification markers
Author(s) -
Eberlein Bernadette,
Hann Rebekka,
Eyerich Stefanie,
Pennino Davide,
Ring Johannes,
SchmidtWeber Carsten B.,
Buters Jeroen
Publication year - 2014
Publication title -
cytometry part b: clinical cytometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.646
H-Index - 61
eISSN - 1552-4957
pISSN - 1552-4949
DOI - 10.1002/cyto.b.21203
Subject(s) - phleum , basophil activation , immunoglobulin e , basophil , population , immunology , antibody , biology , microbiology and biotechnology , chemistry , medicine , environmental health , agronomy
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