【摘要】 目的探讨急性髓系白血病（AML）患者免疫表型和白血病相关的免疫表型（LAIP）特征及在微量残留病（MRD）检测中的意义。方法采用 CIM5/SSC 设门四色流式细胞术,检测610例AML 患者和20名健康志愿者下列抗原的表达:CD7、CD117、CD33、CD34、CD10、CD19、CD56、CD38、CD13、CD14、CD64、CD9、CD16、CD2、CD5、CD11b、CD123、HLA-DR。结果正常骨髓中 CD34⁺和CD117⁺SSC 值低(SSC^low)的细胞比例分别为（0.35±0.15）%和（0.76±0.31）%。CD34⁺SSC^low细胞中绝大多数细胞共表达 CD13、CD33、CD38、CD117、HLA-DR（84%～94%）。CD9和 CD15的相对比例为20%和33%,而 CD11b、CD56、CD19和 CD64的相对比例均低于10%,CD7为12%。CD117⁺SSC^low细胞中,CD19⁺、CD11b⁺、CD56⁺和 CD7⁺细胞相对比例低于10%,CD9⁺细胞为14%,CD38⁺和 HLA-DR⁺细胞的相对比例为87%和91%,其余抗原（CD15、CD13、CD33、CD34）均在30%～50%。AML 患者中 CD117、CD38表达率约为95%,CD33、CD9表达率约为84%。HLA-DR 和 CD13表达率分别为77.23%和75.25%。CD64和 CD34的表达率分别为64.41%和59.51%。CD15表达率为43.06%,CD11b 表达率为22.07%。86.39%的 AML 患者 LAIP 阳性,以 AML-M₁和 M₃最高,AML-M_4Eo最低。LAIP 主要表现为交叉系列抗原表达和非同期共抗原表达,前者以 CD34与 CD7、CD19、CD56同时阳性为主。在非同期抗原表达中,CD34⁺CD64⁺、CD117⁺CD11b⁺、CD117⁺CD38^-/dim、CD117⁺HLA-DR^-/dim正常值在0.01%左右,与 AML 之间的对数差大于3,为灵敏度较高的 LAIP。结论多参数流式细胞术适于80%以上 AML 患者 MRD 检测。更多还原

【Abstract】 Objective To analyze the immunophenotype and leukemia associated immunophenotype （LAIP）of leukemia cells from patients with acute myeloid leukemia（AML）in minimal residual disease （MRD）detection.Methods Four-color multiparametric flow eytometry（FCM）with CD45/SSC gating was used to determine the expression of the following antibodies of CD7,CD117,CD33,CD34,CD10,CD19, CD56,CD38,CD13,CD14,CD64,CD9,CD16,CD2,CDS,CD11b,CD123,HLA-DR in610 AML pa- tients and 20 normal bone marrow（NBM）samples.Results The mean percentages of CD34⁺ and CD117⁺ side scatter low(SSC^low)cells in NBM mononnclear cells（BMMNCs）were（0.35±0.15）% and（0.76± 0.31）%,respectively.The majority（84%-95%）of CD34⁺ SSC^low cells co-expressed CD13,CD33,CD38, CD117 and HLA-DR.33% and 20% of CD34⁺SSC^low cells were CD15⁺ and CD9⁺,respectively.Only a small proportion（<10%）of CD34⁺SSC^low cell co-expressed CD11b,CD56,CD19 and CD64,while co-ex- pressed CD7 was 12%.In CD117⁺SSC^low cells,the relative proportions of CD19⁺,CD11b⁺,CD56⁺ and CD7⁺ were less than 10%,while CD9⁺ was 14%.CD38⁺ and HLA-DR⁺ were 87% and 91%,respectively. The expressions of CD15,CD34,CD13 and CD33 on CD117⁺ SSC^low cells were between 30% and 50%.In AML patients,most cases were CD117⁺（95.08%）,CD38⁺（94.74%）,CD9⁺（84.93%）,CD33⁺ （84.60%）,HLA-DR⁺（77.23%）and CD13⁺（75.25%）.The proportions of CD64⁺ and CD34⁺ were 64.41% and 59.51%,and that of CD15⁺ and CD11b⁺ were 43.06% and 22.07%,respectively.86.39% of AML patients were found to have at least one LAIP,the highest incidence being in AML-M₁ and M₃ sub- types and the lowest in AML-M_4Eo subtype.The cross-lineage antigen and asynchronous antigen expression were the most frequent aberrant phenotypes.CD7,CD19 and CD56 expressing on CD34⁺ cells were major cross-lineage antigen.For asynchronous antigen expression,CD34⁺ CD64⁺,CD117⁺ CD11b⁺,CD34⁺ CD38^-/dim and CD34⁺ HLA-DR^-/dim were seldom expressed on normal BMMNCs（about 0.01%）,and the logarithm difference between AML and NBM was larger than 3.0,being the more sensitive LAIP.Conclusion MRD detection by multiparameter flow cytometry can be applied to more than 80% of AML patients.更多还原