六色流式细胞术用于儿童急性B系淋巴细胞白血病相关免疫表型的检测  

The detection of leukemia-associated immune phenotypes by six-color flow cytometry in pediatric B-lineage acute lymphoblastic leukemia

作　　者：刘光明 吴韶清 孙新 

机构地区：[1]广州市妇女儿童医疗中心,广州510623

出　　处：《中国小儿血液与肿瘤杂志》2012年第3期104-109,共6页Journal of China Pediatric Blood and Cancer

基　　金：广州市医药卫生科技项目(项目编号:2009-YB-078);广东省医学科研基金(项目编号:B2010274)

摘　　要：目的比较六色流式细胞术与三色流式细胞术在急性B系淋巴细胞白血病(B-ALL)中检测出白血病相关免疫表型(LAIP)的差异。方法利用两组六色抗体组合CD20/CD34/CD10/CD22/CD45/CD19、CD7/CD5/CD13/CD33/CD45/CD19对25例三色流式免疫分型诊断为B-ALL的初诊患儿骨髓标本进行检测,通过FSC/SSC、CD45/SSC、CD19/SSC设门,在多个双参数二维点图上对幼稚细胞群、CD19+细胞群进行分析。结果三色流式免疫分型使用23种抗体共检测出CD19strong、CD10strong、CD34strong、CD22strong、CD19+/CD45-/dim+、CD19+/CD34+/CD38-、CD19+/CD13+、CD19+/CD15+、CD19+/CD33+9种LAIP。25例患儿中23例检测出LAIP,其中1个LAIP 4例,2个LAIP 8例,3个LAIP 5例,4个LAIP 3例,5个LAIP 3例。六色流式细胞术使用10种抗体共检测出CD10strong、CD34strong、CD22strong、CD45-/dim+、CD19+/CD13+、CD19+/CD33+、CD19+/CD34+/CD20+、CD19+/CD34+/CD22++、19+/CD34+/CD10dim+、19+/CD45-/dim+/CD34-/dim+、19+/CD10-/CD20-/CD34-11种LAIP。25例中23例检测出LAIP,其中1个LAIP 4例,2个LAIP 5例,3个LAIP 3例,4个LAIP 6例,5个LAIP 5例。结论儿童B-ALL患者具有LAIP者比例较高,适用于运用流式细胞术对B-ALL患儿进行化疗骨髓缓解(CR)后微小残留病(MRD)检测。六色流式细胞术使用的抗体总数较少,但通过FSC/SSC、SSC/CD45、SSC/CD19设门,对幼稚细胞群、CD19+细胞群在多个双参数点图上进行分析,能发现比三色流式细胞术更多的LAIP,并且六色流式细胞术可在同一试管中同时检测6种抗原,能根据患儿初诊时免疫分型结果进行灵活调整,更有利于进行CR后MRD的检测。Objective To compare the differences between three-color and six-color flow cytometry （FCM） on detecting leukemia-associated immune phenotype （LAIP） in pediatric B-lineage acute lymphoblastic leukemia （B-ALL）. Methods Twenty-five patients diagnosed as B-ALL by three-color FCM were detected by two groups of six-color antibody combinations： CD20/CD34/CDlO/CD22/CD45/ CD19 and CD7/CD5/CD13/CD33/CD45/CD19. Naive cell populations and CD19 ＋ cell populations were analyzed on multiparametric two-dimensional point maps through FSC/SSC, CD45/SSC and CD19/SSC gating. Results Nine LAIPs were detected using 23 antibodies by three-color FCM, including CD19strong, CD10 strong , CD34 strong, CD22 strong , CD45 dim ＋/- , CD19 ＋/CD34 ＋/CD38 - , CD19 ＋/CD13 ＋ , CD19＋/CD15＋ and CD19＋/CD33＋. Twenty-three cases （23/25） had LAIPs detected, of which, 4 cases had 1 LAW, 8 cases had 2 LAIPs, 4 cases had 3 LAIPs, 3 cases had 4 LAIPs and 3 had 5 LAIPs. By contrast, 11 LAIPs were detected using 10 antibodies by six-color FCM, including CD10 strong , CD34 strong, CD22 strong , CD45 -/dim, CD19＋/CD13 ＋ , CD19 ＋/CD33 ＋ , CD19 ＋/CD34 ＋/CD20 ＋ , CD19 ＋/CD34 ＋/CD22 ＋ ＋ , 19 ＋/CD34 ＋/CD10 dim＋, 19 ＋/CD45-/dim＋/CD34-/dim ＋ and 19 ＋/CD10-/ CD20-/CD34-. Twenty-three eases （23/25） had LAIPs detected, of which, 4 eases had 1 LAIP, 5 eases had 2 LAWs, 3 eases had 3 LAIPs, 6 eases had 4 LAIPs and 5 had 5 LAIPs. Conclusions High proportions of LAIPs can be detected by FCM in B-ALL patients. Though fewer number of antibodies was used, more LAIPs were detected by six-color FCM than those by three-color FCM. Besides, six-color FCM can detect six antibodies at the same time in one tube, and adjust the antibody combination flexibly, which makes MRD detection more reasonable and convenient.

关 键 词：六色流式细胞术 白血病相关免疫表型 白血病 急性 B细胞 

分 类 号：R733.71[医药卫生—肿瘤]