Sainte-Laudy J, Sabbah A, Vallon C, Guerin J C
Unité d'immuno-allergologie, Laboratoire Cerbra, Val d'Oise, France.
Inflamm Res. 1998 Oct;47(10):401-8. doi: 10.1007/s000110050351.
On the basis of flow cytometric methods previously described for the analysis of human basophil activation, we present here a bi-color anti-IgE FITC, anti CD63 PE method and the correlation with histamine release.
Subjects allergic to grass pollen were selected by their clinical history, skin tests and specific IgE.
Basophils gated in the lymphocyte region of the side scatter (SSC)/forward scatter (FSC) pattern were selected by their high IgE epitope density. Percentage of cells expressing CD63 marker, upregulated on activated basophil membrane, was calculated by the cytometer. Histamine released into the supernatants was measured by RIA.
In these conditions, flow cytometric analysis of blood leukocytes showed that the selected cells had the phenotype CD14-, CD19-, CD45+, IgE++ and CD63- or + which is related to human basophil phenotype, the isotype controls being negative. The use of an anti-CD41 FITC antibody also showed the presence of aggregated platelets on the basophil membrane, CD63 antigen being, however, expressed by basophils themselves and not by platelets. Moreover, no statistical difference was observed between histamine release and flow cytometry after passive sensitization of blood donor leukocytes.
Flow cytometry, as a popular method often used in the immunology and haematology departments of clinical laboratories may represent a new alternative for allergy diagnosis and basophil pharmacology.
基于先前描述的用于分析人类嗜碱性粒细胞活化的流式细胞术方法，我们在此展示一种双色抗IgE FITC、抗CD63 PE方法及其与组胺释放的相关性。
通过临床病史、皮肤试验和特异性IgE选择对草花粉过敏的受试者。
在侧向散射（SSC）/前向散射（FSC）模式的淋巴细胞区域中通过高IgE表位密度选择嗜碱性粒细胞。通过细胞仪计算在活化嗜碱性粒细胞膜上上调表达的CD63标志物的细胞百分比。通过放射免疫分析法测量释放到上清液中的组胺。
在这些条件下，对血液白细胞的流式细胞术分析表明，所选细胞具有CD14 - 、CD19 - 、CD45 + 、IgE ++ 和CD63 - 或 + 的表型，这与人类嗜碱性粒细胞表型相关，同型对照为阴性。使用抗CD41 FITC抗体也显示嗜碱性粒细胞膜上存在聚集的血小板，然而，CD63抗原由嗜碱性粒细胞自身表达而非血小板表达。此外，在对献血者白细胞进行被动致敏后，组胺释放与流式细胞术之间未观察到统计学差异。
流式细胞术作为临床实验室免疫科和血液科常用的一种流行方法，可能代表过敏诊断和嗜碱性粒细胞药理学的一种新选择。
