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在小儿B细胞前体急性淋巴细胞白血病患者中实施用于微小残留病诊断的Ig/TCR靶点识别标准策略:波兰的经验

Implementation of the standard strategy for identification of Ig/TCR targets for minimal residual disease diagnostics in B-cell precursor ALL pediatric patients: Polish experience.

作者信息

Dawidowska Małgorzata, Jółkowska Justyna, Szczepański Tomasz, Derwich Katarzyna, Wachowiak Jacek, Witt Michał

机构信息

Department of Molecular and Clinical Genetics, Institute of Human Genetics, Polish Academy of Sciences, Strzeszyńska 32, 60-479 Poznań, Poland.

出版信息

Arch Immunol Ther Exp (Warsz). 2008 Nov-Dec;56(6):409-18. doi: 10.1007/s00005-008-0045-y. Epub 2008 Dec 1.

Abstract

INTRODUCTION

Minimal residual disease (MRD), detected based on immunoglobulin and T-cell receptor (Ig/TCR) gene rearrangements as markers of residual leukemic cells, is currently the most reliable prognostic factor in acute lymphoblastic leukemia (ALL). A feasibility study is presented of the standard strategy for the identification of Ig/TCR targets for MRD diagnostics in Polish ALL patients by identifying Ig/TCR gene rearrangement pattern using standard primer sets and protocols.

MATERIALS AND METHODS

The PCR-heteroduplex approach based on BIOMED-1 and BIOMED-2 protocols (recommended as the European standard) was used to detect IGH, IGK-Kde, TCRD, TCRG, and TCRB rearrangements in 58 Polish B-cell precursor ALL patients. Sequencing and homology analysis between the obtained and germline Ig/TCR sequences enabled identification of the rearrangements. The U-Gauss test was used for statistical analysis of the Ig/TCR rearrangement pattern in Polish patients compared with relevant data on other nationalities.

RESULTS

The following pattern was identified: IGH: 83% (VH-JH: 74%, DH-JH: 9%), IGK-Kde: 41%, TCRD: 78% (incomplete TCRD: 55%, Vdelta2-Ddelta3: 45%, Ddelta2-Ddelta3: 21%, Vdelta2-Jalpha: 35%), TCRG: 50%, and TCRB: 13%. Considerable convergence of the Ig/TCR pattern in Polish patients and those of other nationalities (mainly West Europeans) was demonstrated. Statistically relevant differences were only found between the incidence of DH-JH in Polish (9%) and Dutch patients (24%; p<0.05) and Polish and Italian patients (19%; p<0.05), VH-JH in Polish (74%) and Chilean patients (100%; p<0.05), and TCRG in Polish (50%) and Brazilian patients (69%; p<0.05).

CONCLUSIONS

The convergence of Ig/TCR patterns in Polish and European patients indicates that the strategy for Ig/TCR target identification based on standard primers and protocols might be directly used for the construction of Polish standards and recommendations for MRD diagnostics.

摘要

引言

基于免疫球蛋白和T细胞受体(Ig/TCR)基因重排作为残留白血病细胞标志物检测到的微小残留病(MRD),是目前急性淋巴细胞白血病(ALL)中最可靠的预后因素。本文介绍了一项可行性研究,该研究通过使用标准引物组和方案识别Ig/TCR基因重排模式,来确定波兰ALL患者MRD诊断中Ig/TCR靶点的标准策略。

材料与方法

采用基于BIOMED-1和BIOMED-2方案(被推荐为欧洲标准)的PCR-异源双链方法,检测58例波兰B细胞前体ALL患者的IGH、IGK-Kde、TCRD、TCRG和TCRB重排。通过对获得的Ig/TCR序列与种系序列进行测序和同源性分析,确定重排情况。使用U-Gauss检验对波兰患者的Ig/TCR重排模式与其他国家相关数据进行统计学分析。

结果

确定了以下模式:IGH:83%(VH-JH:74%,DH-JH:9%),IGK-Kde:41%,TCRD:78%(不完全TCRD:55%,Vdelta2-Ddelta3:45%,Ddelta2-Ddelta3:21%,Vdelta2-Jalpha:35%),TCRG:50%,TCRB:13%。结果表明波兰患者与其他国家(主要是西欧国家)患者的Ig/TCR模式有相当程度的趋同。仅在波兰患者(9%)与荷兰患者(24%;p<0.05)、波兰与意大利患者(19%;p<0.05)的DH-JH发生率之间,波兰患者(74%)与智利患者(100%;p<0.05)的VH-JH发生率之间,以及波兰患者(50%)与巴西患者(69%;p<0.05)的TCRG发生率之间发现了具有统计学意义的差异。

结论

波兰患者与欧洲患者Ig/TCR模式的趋同表明,基于标准引物和方案的Ig/TCR靶点识别策略可能直接用于构建波兰MRD诊断的标准和建议。

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