摘要
目的 建立评价中性粒细胞吞噬和氧化功能的流式细胞仪方法。方法 取健康成人、儿童外周血,佛波 脂(PMA)刺激中性粒细胞活化后吞噬并氧化二氢若丹明123(DHR)为可发出绿色荧光的若丹明123,用流式细胞 仪检测阳性细胞以评价中性粒细胞吞噬和氧化功能。结果10μg／mL佛波脂(PMA)可有效刺激中性粒细胞吞噬 和氧化功能。20例健康成人无刺激时中性粒细胞活化率<10％[(4±3)％],10μg／mL PMA刺激后中性粒细胞活 化率均>90％[(96±3)％];19例健康儿童无刺激时中性粒细胞活化率<10％[(5±4)％],10μg／mL PMA刺激后 中性粒细胞活化率均>90％[(95±4)％];8例足月儿脐带血无刺激时中性粒细胞活化率为(7±4)％,10μg／mL PMA刺激后中性粒细胞活化率(89±6)％。采用刺激指数(SI),PMA刺激与对照荧光强度几何均数,也可用于评 价中性粒细胞刺激后活化情况,但变异较大。结论 流式细胞仪-DHR分析可用于临床评价成人与儿童中性粒 细胞吞噬和氧化功能。
Purpose To establish a method for evaluating phagocyte and oxidation function (respiratory burst) of neutrophils by flow cytometry.Methods Intracellular oxidation of dihydrorhodamine 123(DHR) to the fluorescent compound rhodamine 123 (Rhol23) was used to detect the production of oxygen metabolites in phorbol myristate acetate (PMA) activated neutrophils. Peripheral blood samples were got from 20 healthy adults and 19 healthy children, and 8 cord blood samples were got. Results We found (hat 10 uuuuuuuuuug/mL PMA was effective for the activation of neutrophils. In 20 adult cases, the ratio of activated neutrophils was less than 10%[(4+3)%] in nan-stimulated tubes. The average activated ratio was more than 90 %[ (96+3) % ] in PMA stimulated tubes. Similar results were found in children,activated neutrophils were less than 10%[(5+4)%] in non-stimulated test and more than 90% [ (95+4) % ] in PMA treated tubes. Activated neutrophils were (7+4) % in non-stimulated and (89+6) % in PMA stimulated cord blood. The variance was considerable when stimulated index (SI) was used to evaluate the function of neutrophils that based on geometric mean of fluorescence intensity.Conclusions Flow cytometry-DHR assay could be used for evaluate neutrophl function in clinic for adults as well as children.
出处
《复旦学报（医学版）》
CAS
CSCD
北大核心
2005年第1期101-104,共4页
Fudan University Journal of Medical Sciences
