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目的探讨利用四色流式细胞术(FCM)检测 B 细胞急性淋巴细胞白血病(B-ALL)微量残留病(MRD)的临床意义。方法采用以抗 CD34/CD10/CD45/CD19为主的两种四色荧光标记抗体组合,对98例 B-ALL 患者的671份骨髓标本和1份脑脊液标本进行 FCM 多参数 MRD 检测,98例随访患者中26例无发病初期的免疫分型资料。结果 FCM 检测显示白血病细胞<0.0001(MRD 阴性)的标本为579份;白血病细胞>0.0001的样本数为93份,其中64份骨髓标本白血病细胞比例<0.05,29份标本白血病细胞比例>0.05,我们将其归为复发病例(包括治疗后未达 CR 的病例)。共20例患者复发,其中19例血液学复发,1例中枢神经系统复发。15例血液学复发者在复发前7～17周发现MRD 阳性,包括发病时免疫分型资料不明者6例,MRD 水平均>0.0001;2例分别在复发前3个月和9个月检测 MRD 为阴性,此后中断检测。在诱导治疗结束和治疗3个月时,如果 MRD 水平>0.0001,复发率为50%(12例中有6例复发),而 MRD 阴性组复发率为7.5%(40例中有3例复发)(P=0.000)。结论利用 FCM 进行 MRD 跟踪临测可预测复发,治疗初期患者 MRD>0.0001,复发的危险性较高。而在掌握了正常 B 祖细胞抗原表达规律的基础上,可不完全依赖于发病时的免疫表型资料。
Objective To investigate the clinical significance of using four-color flow cytometry (FCM) to detect micro-residual disease (MRD) in B-cell acute lymphoblastic leukemia (B-ALL) Methods A total of 671 bone marrow specimens and 1 cerebrospinal fluid from 98 patients with B-ALL were examined by FCM multi-parameter MRD with two anti-CD34 / CD10 / CD45 / CD19 two-color fluorescent labeled antibodies. 26 cases of patients without early onset of immunophenotyping data. Results FCM showed that the number of leukemia cells with <0.0001 (MRD negative) was 579 and the number of leukemia cells with 0.0001 was 93. Among them, the proportion of 64 myeloid leukemia cells was less than 0.05 and the percentage of leukemia cells was over 0.05. It is classified as a recurrent case (including cases of non-CR after treatment). A total of 20 patients relapsed, of which 19 cases of hematological relapse, 1 case of central nervous system recurrence. In 15 hematological relapse patients, MRD was positive 7 to 17 weeks before relapse, including 6 patients with unknown immunophenotype at onset and MRD> 0.0001; 2 patients were examined MRD at 3 months and 9 months before recurrence Negative, then cut off the test. At the end of induction therapy and at 3 months of treatment, the recurrence rate was 50% (MRD level> 0.0001) (6 of 12 patients relapsed) and 7.5% of MRD-negative patients (3 of 40 patients relapsed) (P = 0.000). Conclusion MRD tracing using FCM can predict the recurrence, the initial treatment of patients with MRD> 0.0001, the higher the risk of recurrence. On the basis of grasping the regular expression of antigen of B progenitor cells, it may not be completely dependent on the immunophenotype data at onset.