造血B祖细胞与急性B淋巴细胞白血病细胞的流式细胞术免疫表型分析与鉴别  

Immunophenotypic analysis and identification of hematogones and acute B lymphoblastic cells by flow cytometry

作　　者：许艳丽[1] 王顺清[1] 杜庆华[1] 谢健晋[1] 

机构地区：[1]广州市第一人民医院血液科,510180

出　　处：《白血病．淋巴瘤》2015年第5期282-286,共5页Journal of Leukemia & Lymphoma

摘　　要：目的 分析造血B祖细胞与急性B淋巴细胞白血病（B-ALL）细胞在形态学、免疫表型等方面的差异,为造血B祖细胞、微小残留白血病细胞的正确鉴定提供参考.方法 采用流式细胞术对临床确诊的58例B-ALL患者初诊、缓解、复发期的132份骨髓样本进行细胞免疫表型分析,通过CD34/CD10/CD19/CD45或CD34/CD10/CD45/CD19/CD20/CD38抗体组合鉴定造血B祖细胞群.结果 132份骨髓样本中,45份（34％）检出造血B祖细胞群,检测范围0～36％,其中3份样本出现在初诊病例,1份样本出现在复发病例,41份样本出现在缓解病例.66份缓解病例样本中,造血B祖细胞检出率为62％（41/66）.白血病细胞在初诊、复发病例中均占有核细胞群体5％以上,24份缓解病例样本可检出低于5％的残留白血病细胞.造血B祖细胞与白血病细胞共存在28份样本中,其中3份出现在初诊病例,1份出现在复发病例,24份出现在缓解病例.造血B祖细胞CD45呈阴性至弱阳性连续分布,侧向散射信号（SSC）较低,早期造血B祖细胞群CD34＋,随着细胞成熟度的增加,CD34消失.CD19、CD10在造血B祖细胞整个阶段均为阳性,早期CD10表达强度更高.白血病细胞呈“发育阻滞”模式,呈现为较均一的荧光细胞群;SSC值较正常B祖细胞偏高.白血病细胞抗原表达过强或过低的表型在正常造血B祖细胞均未见到,且造血B祖细胞不会出现交叉系列抗原标记,CD20呈阴性至弱阳性的连续分布.结论 造血B祖细胞在B-ALL患者不同阶段均不同程度地存在,尤其在化疗后的骨髓恢复期有一过性增长,分析此阶段的样本鉴定B系群体来源需谨慎.了解正常细胞的发育背景、认知白血病细胞表型的漂移变化模式及掌握多色分析优化抗体组合的流式细胞技术,对白血病细胞的准确鉴定及微小残留病检测的准确性提升具有重要意义.Objective To discriminate morphology and immunophenotype differences between hematogones and lymphoblast to provide some references for the correct identification of hematogones and minimal residual leukemia cells.Methods Immunophenotypes were detected by flow cytometry in a total of 132 bone marrow from 58 patients with acute B lymphoblastic leukemia during diagnosis,remission and relapse.Hematogones were identified based on combination of CD34/CD10/CD19/CD45 or CD34/CD10/CD45/CD19/CD20/CD38.Results Among 132 specimens,45 （34 %） were identified hematogones,the detection range was 0-36 %.Three specimens appeared in diagnosis patients,one in relapse,and the remaining 41 cases in remission.The detection rate of hematogones was 62 % （41/66） in the remission cases.More than 5 % leukemia cells of nucleated cells were detected in diagnosis and relapse,and less than 5 % residual leukemia cells was in 24 specimens from remission patients.In 28 specimens,the co-existence of hematogones and leukemia cells was found,including three in diagnosis,one in relapse and the remaining 24 in remission.Hematogones were characterized in term of variable expression of CD45 and very low side scatter.The early hematogones expressed CD34.With maturation increasing,hematogones gradually lacked CD34.CD19 and CD10 were presented in whole hematogones stage.Early hematogones had expression of CD10.Lymphoblasts showed maturation arrest and more homogeneous populations.SSC values of hematogones were higher than that of normal B cell progenitors.Antigen overexpression or underexpression was not found in normal hematopoietic progenitor B cells,and hematopoietic progenitor B cells did not appear cross-lineage markers,CD20＋ cells exhibited continuous distribution from negative to weak positive for normal hematogones.Conclusions Hematogones were present in diagnosis,remission and relapse cases with acute B lymphoblastic leukemia,especially abundant in bone marrow after chemotherapy.It should be careful to diagnose and discriminate the malig

关 键 词：急性B淋巴细胞白血病 造血B祖细胞 多参数流式细胞术 免疫分型 

分 类 号：R733.71[医药卫生—肿瘤]