摘要
目的 :应用流式细胞术 ,建立一种准确、外周血嗜酸性细胞及其相关分子的快速测定方法。方法 :采集正常人外周血 ,用茶碱 (10 -4mol/L)、地塞米松 (10 -4mol/L)和rhIL 5 (10 -8mol/L)预处理 ,用抗CD16 PEmAb与FITC标记的抗相关细胞分子的进行双标记染色 ,并以CD16 FL2辅助设门 ,准确找到嗜酸性粒细胞群 ,然后对其相关分子进行分析。结果 :嗜酸性粒细胞定位准确 ,茶碱和地塞米松能够抑制IL 5引起的Eos的活化并使其表面的CD6 2L脱落。结论 :应用流式细胞术与二色荧光mAbCD16 PE阴性细胞法设门 ,可准确快速地检测外周血中嗜酸性粒细胞及分子的表达率 ,血液用样量小 ,人为影响因素少 ,是免疫学基础研究和临床检验较理想的测定方法。
AIM: To set up an accurate and rapid method to detect the phenotype of peripheral blood eosinophils using flow cytometry. METHODS: 10 -4 mol/L of theophylline, 10 -4 of dexamethasone and 10 -8 mol/L of rhIL-5 were added to peripheral blood sampled from normal human subjects. Anti-CD16-PE mAb and FITC-labelled mAbs to eosinophil’s molecules were used to perform double labeling staining. Simultaneously, CD16-FL2 was used to install gating so as to accurately locate eosinophils. And then their molecules were examined and analyzed. RESULTS: The eosinophils were accurately located. Theophylline and dexamethasone effectively inhibited activation of eosinphils and sheding of CD62L on eosinophils caused by IL-5. CONCLUSION: Our method can accurately detect eosinophils in peripheral blood samples and their molecules. In addition, only a tiny amount of blood sample is needed and few artificial factors are involved in the detection. Therefore, this method may be an ideal both in basic immunological research and clinical laboratory examination.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2004年第4期478-480,共3页
Chinese Journal of Cellular and Molecular Immunology
