三组四色抗体的流式细胞术监测儿童B系急性淋巴细胞白血病微小残留病  

Flow Cytometry Screening for Minimal Residual Disease in Childhood B Lineage Acute Lymphoblastic Leukemia by 3 Panels of 4-Color Antibodies

作　　者：张剑波[1] 李戈[2] 母得志[3] 房俊[1] 陈杰[1] 景清[2] 屈艺[3] 

机构地区：[1]四川省人民医院中心实验室 [2]四川省人民医院儿科,成都610072 [3]四川大学华西第二医院儿科

出　　处：《中华妇幼临床医学杂志（电子版）》2008年第4期10-14,共5页Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition)

基　　金：国家自然科学基金项目(编号:30770748);教育部博士点基金项目(编号:20050610094)~~

摘　　要：目的建立一种敏感的三组四色抗体流式细胞术,监测儿童B系急性淋巴细胞白血病(B lineage acute lymphoblastic leukemia,B-ALL)微小残留病(minimal residual disease,MRD),并探讨其应用价值。方法采用三组四色荧光抗体组合(CD_(45)/CD_(19)/CD_(10)/CD_(34),CD_(45)/CD_(19)/CD_(10)/CD_(13)和CD_(45)/CD_(19)/CD_(22)/CD_(34))检测18份初诊为B系急性淋巴细胞白血病患儿的骨髓样本(A组)和67份该病治疗后缓解患儿骨髓样本(B组)的白血病相关异常免疫表型(leukemia-associated aberrant immunophenotyping,LAIP)细胞。A,B两组样本来源于2004年6月至2007年12月在四川省人民医院儿科住院和定期随访的42例患儿,年龄为1～12岁;男性患儿为30例,女性为12例。同时,检测12份正常儿童骨髓(C组)作为对照(同期在本院就诊)。结果由于B系急性淋巴细胞白血病患儿骨髓的白血病相关异常免疫表型细胞与正常骨髓B细胞,在流式散点图中分布位置有差异,从而筛选出8个有价值的双参数散点图,以监测临床缓解B系急性淋巴细胞白血病患儿的微小残留病。本方法在B系急性淋巴细胞白血病A组和B组患儿骨髓中的微小残留病阳性检出率分别为89%(16/18)和48%(32/67);白血病相关异常免疫表型细胞在A,B两组样本中分别为(67.8±36.4)%((?)±s)和(0.056±0.083)%((?)±s),A组患儿均明显高于B组患儿(P<0.01)。分析3例复发患儿复发前的跟踪监测结果发现,复发前均检测到微小残留病呈阳性,并且其白血病呈相关异常免疫表型细胞均明显升高。结论本研究建立了一种灵敏、快速、易于标准化操作的监测B系急性淋巴细胞白血病患儿微小残留病的方法。Objective Minimal residual disease （MRD）in childhood B lineage acute lymphoblastic leukemia （B ALL） is an important indicator for treatment and prognosis. We set up a flow cytometric method with three panels of 4-color antibodies to enhance detection sensitivity and test its clinical application. Methods Three panels of 4 color fluorochrome monoclonal antibodies （CD45/CD19/CD10/ CD84, CD45/CD19/CD10/CD13 and CD45/CD19/CD22/CD34） were used to detect the leukemia-associated aberrant immunophenotyping （LAIP） cells in 18 bone morrow samples of newly diagnosed B-ALL patients （the group A） and 67 bone morrow samples of clinical remission patients（the group B）. All the cases were inpatients or scheduled follow-up patients registered in the Department of Pediatrics, Sichuan Provincial People＇s Hospital from June 2004 to December 2007. Meanwhile,Bcells of 12 normal bone marrows were used as controls （the groupC）. Results Eight panels of the 2-parameter dot plots to monitor the minimal residual disease in the clinical remission patients were set up to tell leukemia-associated aberrant immunophenotype cell populations of B lineage acute lymphoblastic leukemia from the normal. The positive detection rate for minimal residual disease was 89%（16/18） in the group A and 48%（32/67） in the group B （P〈0. 01）. The average leukemia associated aberrant immunophenotype cell levels of the groupA （67.8±36.4）% （x±s）wereobviousiy higher than those of the group B（0.056±0.083）%（x±s） with statistical significance （P〈0.01）. By analyzing the leukemia-associated aberrant immunophenotype cell results of 3 relapsed patients with regular follow-up, all of them were found minimal residual disease positive at remission stage, and their leukemia associated aberrant immunophenotype cell level rose sharply immediate before recurrence. Conclusion The method described here was a sensitive and rapid method for monitoring minimal residual disease in children with B lineage acute lymphobla

关 键 词：微小残留病 白血病 B细胞 急性 流式细胞术 

分 类 号：R733.7[医药卫生—肿瘤]