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目的探讨流式细胞仪在急性粒细胞白血病(acute myeloid leukemia,AML)免疫分型中的特点和规律。方法利用流式细胞术中CD45/SSC双参数散点图设门法,对四色荧光素(FITC、PE、PC5、ECD)标记的骨髓血细胞免疫表型分析。取初发患者骨髓单细胞悬液100μl加入到测试管中,再加入特定抗体,室温避光1 h,然后经溶血处理,上机检测。每管均设置同型对照。结果在76例AML中,髓系相关抗原MPO、CD33、CD65表达最高,总阳性率分别为96.1%、94.7%和93.4%,CD13、CD15、CD16的表达率较低,总阳性率分别为50.0%、63.2%和89.5%;CD14、CD64在M4、M4EO、M5中阳性率最高;CD41、CD61在M7中的阳性率最高;CD34在M1和部分M2中强表达,但总阳性率约为46.1%。结论本研究成功分析不同亚型AML的CD45/SSC散点图异常细胞分布的特定位置,同时结合各种不同系列分化特异性抗原的表达综合分析,实现与FAB法在AML分型比较符合率高达97%。
Objective To investigate the characteristics and rules of flow cytometry in the immunophenotyping of acute myeloid leukemia (AML). Methods The immunophenotypic analysis of bone marrow blood cells labeled with four-color fluorescein (FITC, PE, PC5, ECD) was performed by CD45 / SSC double-parameter scatter plot flow cytometry in flow cytometry. Take fresh bone marrow mononuclear cell suspension 100μl added to the test tube, and then add specific antibodies, dark at room temperature 1 h, and then hemolysis treatment, the machine test. Each tube is set the same type of control. Results The expression of MPO, CD33 and CD65 in myeloid-associated antigen was the highest in 76 cases of AML with the total positive rates of 96.1%, 94.7% and 93.4%, respectively, and the positive rates of CD13, CD15 and CD16 were lower %, 63.2% and 89.5%, respectively. The positive rates of CD14 and CD64 in M4, M4EO and M5 were the highest. The positive rates of CD41 and CD61 in M7 were the highest. CD34 was strongly expressed in M1 and M2, but the positive rate was 46.1 %. Conclusions This study successfully analyzed the specific location of abnormal cell distribution of CD45 / SSC scattergram in different subtypes of AML. Combined with the expression analysis of different series of differentiation-specific antigens, the coincidence rate with FAB in AML typing was higher 97%.