Bridts Chris H, Sabato Vito, Mertens Christel, Hagendorens Margo M, De Clerck Luc S, Ebo Didier G
Department of Immunology and Allergology, University of Antwerp, Campus Drie Eiken, D.T.596, Universiteitsplein 1, 2610, Wilrijk, Belgium.
Methods Mol Biol. 2014;1192:147-59. doi: 10.1007/978-1-4939-1173-8_11.
The basis of flow cytometric allergy diagnosis is quantification of changes in 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 up-regulate 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 detection of surface marker activation (Method 1) and changes in intragranular histamine (Method 2), both reflecting different facets of basophil activation.
流式细胞术诊断过敏的基础是对嗜碱性表面膜标志物表达变化进行定量分析（Ebo等人，《临床与实验过敏》34: 332 - 339，2004年）。当遇到由表面受体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并产生可定量的生物活性介质，还会上调不同标志物（如CD63、CD203c）的表达，这些标志物可通过使用特异性单克隆抗体的多色流式细胞术进行检测（Ebo等人，《细胞分析B：临床细胞分析》74: 201 - 210，2008年）。在此，我们描述了两种基于流式细胞术的方案，它们能够检测表面标志物激活（方法1）和颗粒内组胺的变化（方法2），二者均反映了嗜碱性粒细胞激活的不同方面。
