Optimizing of the basophil activation test: Comparison of different basophil identification markers.
نویسندگان
چکیده
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.
منابع مشابه
Expression of Basophil Activation Markers in Pediatric Asthma
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Dear Editor, We are grateful to Dr. Chirumbolo for the letter in response to our paper describing a basophil activation test for chronic urticaria [1, 2]. We agree with most of the issues raised by Dr. Chirumbolo, but some points need to be addressed. The non-atopic donors in our study were adults, not infants. Among 3-5 healthy individuals with blood group O, one with a high basophil count was...
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عنوان ژورنال:
- Cytometry. Part B, Clinical cytometry
دوره 88 3 شماره
صفحات -
تاریخ انتشار 2014