异常免疫表型模式在急性髓系白血病微小残留检测中的应用

目的用多参数流式细胞术区分急性髓性白血病细胞与正常骨髓原始细胞免疫表型,建立急性髓系白血病细胞异常免疫表型模式和流式细胞术检测急性白血病微小残留病的方法。方法选择正常骨髓标本和急性髓性白血病患者骨髓标本,用流式细胞术分析髓系原始细胞免疫表型特征。并将标记抗原用于分析初发白血病细胞免疫表型,以寻找急性髓系白血病细胞异常免疫表型模式和确定用于微小残留病检测的抗体组合,更好地提示患者复发情况。并将流式细胞术结果、骨髓细胞形态学结果和PCR融合基因检测结果进行比较。结果分析20例非恶性血液系统恶性疾病患者的骨髓标本中髓系原始细胞的免疫表型,CD117阳性髓系原始细胞占2.51%±0.84%,CD34阳性髓系原始细胞占1.21%±0.83%,均有特定免疫表型特征。检测346例非早幼粒细胞白血病的急性髓性白血病患者免疫表型特征,异常免疫表型有314例,占90.80%,其中跨系表达抗原主要包括CD7(31.21%)、CD56(23.70%)、CD19(13.87%),跨阶段共表达的抗原包括CD34/CD64(2.89%)、CD14/CD117(1.16%),抗原表达强度变化或抗原缺失主要包括原始粒细胞不表达HLA-DR(4.62%)、CD13(7.51%)或CD33(12.14%),以及单核细胞不表达CD14(5.20%)。早幼粒细胞白血病患者31例,有异常免疫表型的仅占16.7%,低于非早幼粒细胞白血病的急性髓性白血病。根据白血病异常免疫表型模式,与PCR结果符合度高,能很好地提示白血病进展。结论急性髓性白血病细胞具有明显不同于正常来源原始细胞的异常免疫表型特征,适合采用流式细胞术进行微小残留病检测,并能反映体内白血病细胞负荷和病情变化。

Detection of minimal residual disease in acute myeloid leukemia by aberrant immunophenotype

Objective The immunophenotypes of acute myeloid leukemia cells and normal bone marrow primordial cells were distinguished by multiparameter flow cytometry.The abnormal immunophenotype pattern of acute myeloid leukemia cells was established and the flow cytometry was used to detect minimal residual disease of acute leukemia.Methods The immunophenotypic characteristics of myeloid primordial cells were analyzed by flow cytometry.In order to find out the abnormal immunophenotype pattern of acute myeloid leukemia cells and to determine the antibody combination for detection of minimal residual disease,we also used the marker antigen to analyze the immunophenotype of primary leukemia cells.The results of flow cytometry,morphology of bone marrow cells and PCR fusion gene were compared.Results The immunophenotype of myeloid primordial cells in bone marrow of 20 patients with non hematologic malignancies was analyzed.CD117 positive myeloid cells accounted for 2.51%±0.84%,CD34 positive myeloid cells accounted for 1.21%±0.83%,all of which had specific immunophenotypic characteristics.Aberrant immunophenotype were found in 314 of 346 AML(90.80%),acute promyelocytic leukemia excluded.Main immunophenotypes of lineage cross-expressing included CD7(31.21%),CD56(23.70%)and CD19(13.87%).Cross-stage expressing antigen included CD34/CD64(2.89%),CD14/CD117(1.16%).Antigen intensity change or deficiency mainly included HLA-DR(4.62%)、CD13(7.51%),CD33(12.14%)and CD14 on monocytic leukemia(5.20%).In 31 cases of promyelocytic leukemia,only 16.7%had abnormal immunophenotype,which was lower than that of non promyelocytic leukemia.According to the abnormal immunophenotypic pattern of leukemia,it is highly consistent with PCR results,which can well indicate the progression of leukemia.Conclusion Acute myeloid leukemia cells have abnormal immunophenotypic characteristics which are different from those of normal primordial cells.It is suitable to detect minimal residual disease by flow cytometry,and it can reflect the load of leukemia cells and the change of disease condition in vivo.