Optimizing of the Basophil Activation Test: Comparison of Different Basophil Identification Markers
Optimizing of the Basophil Activation Test: Comparison of Different Basophil Identification Markers
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DOI：
10.1002/cyto.b.21203
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发表时间：
2015-05-01
影响因子：
3.4
通讯作者：
Buters, Jeroen
中科院分区：
文献类型：
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作者：
Eberlein, Bernadette;Hann, Rebekka;Buters, Jeroen
背景流式细胞术鉴定嗜碱性粒细胞是检测嗜碱性粒细胞在IgE依赖性或非依赖性刺激下活化的前提。本研究的目的是比较不同的标志物组合在一个同步的biflycocytometric measurement.MethodsTen患者与草花粉过敏和三个对照组被列入研究。通过使用抗CCR 3、抗IgE、抗CRTH 2、抗CD 203 c和抗CD 3对嗜碱性细胞进行门控。细胞被单克隆抗Fc β RI抗体、N-甲酰基-甲硫氨酰基-亮氨酰基-苯丙氨酸（fMLP）和过敏原提取物Phalopratense激活。结果抗CCR 3（+）细胞、抗IgE（+）细胞和抗IgE（+）/抗CD 203 c（+）细胞的嗜碱性粒细胞相对数最高，CRTH 2（+）/CD 203 c（+）/CD 3（-）细胞的嗜碱性粒细胞相对数最低。在所有实验中观察到CCR 3（+）细胞与CCR 3（+）/CD 3（-）细胞和CRTH 2（+）/CD 203 c（+）/CD 3（-）细胞非常好和良好的一致性。CCR 3（+）群体与CD 3（+）细胞的污染以及IgE（+）-群体与CCR 3（-）细胞和CD 203（-）细胞的污染与所有其它标记物组合相比是最低的。这些标记通常可推荐用于鉴定嗜碱性粒细胞。如果需要非常高纯度的嗜碱性粒细胞群体，则应选择抗IgE。(c)2014年国际临床细胞计数学会
BackgroundFlowcytometric 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.MethodsTen 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-Fc epsilon RI antibody, N-formyl-methionyl-leucyl-phenylalanine (fMLP), and the allergen extract Phleum pratense. The activation marker anti-CD63 was used.ResultsThe 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.ConclusionsAs 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. (c) 2014 International Clinical Cytometry Society