Department of Rheumatology, Clinical Immunology and Allergology, Inselspital, University Bern, Bern, Switzerland.
Allergy. 2011 Jan;66(1):85-91. doi: 10.1111/j.1398-9995.2010.02431.x.
Basophil activation tests (BAT) rely on different combinations of basophil selection and activation markers. Whereas activation markers, especially CD63, are widely validated, the most suitable and robust marker for basophil selection is still a matter of debate.
Comparison of cell surface expression of two commonly used basophil selection markers (IgE, CD123/HLA-DR) with CCR3 in an unselected group of atopic and nonatopic donors in resting and activated basophils.
EDTA blood of 94 healthy adults, about half of them atopic by history, was analyzed using two different staining strategies: anti-CD123-PE/anti-HLA-DR-PerCP/anti-lin1-FITC and anti-IgE-FITC/anti-CD3-PerCP/anti-CCR3-PE. Additionally 40 pollen-allergic patients were recruited for the assessment of CCR3 expression after basophil activation.
In resting basophils, cell surface expression of the three basophil selection markers was most constant for CCR3. IgE gating strategy showed the highest variation and up to 80% of nonbasophils in the selected gate in certain donors. During basophil activation, a shift of the mean fluorescence intensity for CCR3 toward the lower third of the CCR3-positive population could be demonstrated, but neither were CCR3-positive cells significantly lost for further analysis nor was differentiation between CCR3-positive and CCR3-negative cell populations hampered by this shift.
CCR3 is a stable and highly expressed basophil selection marker, independent of the atopic background or basophil activation state and allows an accurate identification of basophils without need of a second marker. The basophil markers CD123/HLA-DR and IgE showed significantly higher interindividual variability in cell surface expression and are therefore less suited as selection markers.
嗜碱性粒细胞激活试验（BAT）依赖于不同的嗜碱性粒细胞选择和激活标志物组合。虽然激活标志物，特别是 CD63，已经得到了广泛的验证，但最适合和最稳健的嗜碱性粒细胞选择标志物仍存在争议。
比较两种常用的嗜碱性粒细胞选择标志物（IgE、CD123/HLA-DR）与 CCR3 在未选择的特应性和非特应性供体的静息和激活嗜碱性粒细胞中的细胞表面表达。
使用两种不同的染色策略分析 94 名健康成年人的 EDTA 血液，其中约一半有特应性病史：抗-CD123-PE/抗-HLA-DR-PerCP/抗-lin1-FITC 和抗-IgE-FITC/抗-CD3-PerCP/抗-CCR3-PE。此外，还招募了 40 名花粉过敏患者，以评估嗜碱性粒细胞激活后 CCR3 的表达。
在静息嗜碱性粒细胞中，三种嗜碱性粒细胞选择标志物中，CCR3 的细胞表面表达最稳定。IgE 门控策略显示，在某些供体中，所选门内高达 80%的非嗜碱性粒细胞。在嗜碱性粒细胞激活过程中，可以观察到 CCR3 的平均荧光强度向 CCR3 阳性群体的较低三分之一转移，但没有进一步分析的情况下 CCR3 阳性细胞明显丢失，也没有因这种转移而妨碍 CCR3 阳性和 CCR3 阴性细胞群体的分化。
CCR3 是一种稳定且高表达的嗜碱性粒细胞选择标志物，与特应性背景或嗜碱性粒细胞激活状态无关，可无需使用第二种标志物准确识别嗜碱性粒细胞。嗜碱性粒细胞标志物 CD123/HLA-DR 和 IgE 的细胞表面表达个体间差异明显更大，因此不太适合作为选择标志物。
