CD58在儿童急性B淋巴细胞白血病中的表达及其用于微小残留病检测的可行性研究

Expression of CD58 in childhood B-lineage acute lymphoblastic leukemia and its feasibility in minimal residual disease detection

摘要:

目的 检测淋巴细胞功能相关抗原-3(CD58)在儿童急性B淋巴细胞白血病(B-ALL)中的表达情况,并探讨其用于儿童B-ALL微小残留病(MRD)检测的可行性。方法 选取2014年1~9月确诊为B-ALL的87例患儿为研究对象,同期选取20例非肿瘤、非血液病且骨髓涂片细胞形态正常的住院患儿作为对照组。应用四色流式细胞术(FCM)检测对照组及B-ALL患儿初诊时、诱导化疗15 d时骨髓标本的CD58表达水平。在诱导化疗33 d时,采用实时荧光定量聚合酶链反应(RT-PCR)和FCM对B-ALL患儿进行MRD检测。结果 87例B-ALL患儿骨髓CD58平均荧光强度(MFI)(91±33)高于对照组(14±6)(P< 0.01);其中44例B-ALL患儿CD58强表达。在诱导化疗过程中,诱导化疗15 d时B-ALL患儿骨髓CD58的表达量(105±22)与初诊时(107±26)比较差异无统计学意义(P> 0.05)。对44例CD58强表达B-ALL患儿进行MRD检测,FCM检测MRD阳性患儿9例,阴性患儿35例;RT-PCR检测MRD阳性患儿11例,阴性患儿33例,42例(95%)检测结果一致,两种检测方法检测MRD差异无统计学意义(P> 0.05)。结论 CD58在B-ALL患儿中高表达且表达稳定,可以作为B-ALL患儿MRD检测的指标。

Abstract:

Objective To measure the expression of lymphocyte function-associated antigen-3 (CD58) in childhood B-lineage acute lymphoblastic leukemia (B-ALL) and to explore the feasibility of CD58 as an indicator for minimal residual disease (MRD) detection in childhood B-ALL. Methods Eighty-seven children diagnosed with B-ALL between January 2014 and September 2014 were enrolled, and 20 hospitalized children who had no tumor or blood disease and had normal bone marrow cell morphology served as the control group. The expression features of CD58 in bone marrow samples from the two groups (at diagnosis, on day 15 of induction chemotherapy) were analyzed by four-color flow cytometry (FCM). Quantitative real-time polymerase chain reaction (qRT-PCR) and FCM were used to detect MRD in B-ALL patients on day 33 of induction chemotherapy. Results The mean fluorescence intensity of CD58 expression in the 87 B-ALL cases (91±33) was significantly higher than that in the 20 controls (14±6) (P< 0.01); CD58 was over-expressed in 44 of the B-ALL cases. In the B-ALL children, the expression of CD58 on day 15 of induction chemotherapy (105±22) was not significantly different from that at diagnosis (107±26) (P> 0.05). In the 44 B-ALL patients with CD58 over-expression, FCM showed 9 MRD(+) cases and 35 MRD(-) cases, while qRT-PCR showed 11 MRD(+) cases and 33 MRD(-) cases; 42 cases (95%) showed consistent results of the two tests, so there was no significant difference between the two methods in detecting MRD (P> 0.05). Conclusions CD58 is over-expressed and stable in children with B-ALL, and it can be considered as an indicator for MRD detection in childhood B-ALL.