Department of Immunology, Allergology, Rheumatology, University of Antwerp, Antwerp University Hospital, Antwerp, Belgium.
Faculty of Medicine and Health Science, Department of Immunology, Allergology, Rheumatology and the Infla-Med Centre of Excellence, University of Antwerp, Antwerp University Hospital, Antwerp, Belgium.
Methods Mol Biol. 2020;2163:183-195. doi: 10.1007/978-1-0716-0696-4_15.
The basis of flow cytometric allergy diagnosis is the quantification of changes in the expression of basophilic surface membrane markers (Ebo et al., Clin Exp Allergy 34: 332-339, 2004). Upon encountering specific allergens recognized by surface receptor FcεRI-bound IgE, basophils not only secrete and generate quantifiable bioactive mediators but also upregulate the expression of different markers (e.g., CD63, CD203c) which can be detected by multicolor flow cytometry using specific monoclonal antibodies (Ebo et al., Cytometry B Clin Cytom 74: 201-210, 2008). Here, we describe two flow cytometry-based protocols which allow the detection of surface marker activation (Method 1) and changes in intragranular histamine (Method 2), both reflecting different facets of basophil activation.
流式细胞术过敏诊断的基础是定量检测嗜碱性表面膜标志物的表达变化（Ebo 等人，Clin Exp Allergy 34: 332-339, 2004）。当遇到被表面受体 FcεRI 结合的 IgE 识别的特定过敏原时，嗜碱性粒细胞不仅会分泌和产生可量化的生物活性介质，还会上调不同标志物的表达（例如，CD63、CD203c），这些标志物可以通过使用特异性单克隆抗体的多色流式细胞术检测到（Ebo 等人，Cytometry B Clin Cytom 74: 201-210, 2008）。在这里，我们描述了两种基于流式细胞术的方案，可用于检测表面标志物的激活（方法 1）和细胞内组胺的变化（方法 2），这两种方法均反映了嗜碱性粒细胞激活的不同方面。
