Dawidowska Małgorzata, Jółkowska Justyna, Szczepański Tomasz, Derwich Katarzyna, Wachowiak Jacek, Witt Michał
Department of Molecular and Clinical Genetics, Institute of Human Genetics, Polish Academy of Sciences, Strzeszyńska 32, 60-479 Poznań, Poland.
Arch Immunol Ther Exp (Warsz). 2008 Nov-Dec;56(6):409-18. doi: 10.1007/s00005-008-0045-y. Epub 2008 Dec 1.
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.
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.
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).
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.
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。
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。
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。
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。