Detection of minimal residual disease in acute leukemia by immunological marker analysis and polymerase chain reaction.
Detection of minimal residual disease in acute leukemia by immunological marker analysis and polymerase chain reaction.
复制标题
通过免疫标记分析和聚合酶链反应检测急性白血病微小残留病。
作者：
J. J. V. Dongen;T. Breit;H. Adriaansen;Auke Beishuizen;H. Hooijkaas
微小残留病（MRD）的检测可用于适应或分层急性白血病患者的治疗，并最终导致个性化的治疗方案。虽然白血病细胞通常具有与其正常对应物相当的免疫表型，但基于以下假设，可以使用免疫标记物分析来检测MRD：在其正常繁殖部位和“归巢区域”之外存在阳性细胞指示恶性肿瘤。该方法可用于检测终末脱氧核苷酸转移酶（TdT）阳性T细胞急性淋巴细胞白血病（ALL）患者和TdT+急性髓性白血病（AML）患者的血液和骨髓中以及TdT+白血病患者的脑脊液中的MRD。在其他类型的急性白血病中，免疫标记分析通常不能检测到低频率的恶性细胞，但在其中一部分中，聚合酶链反应（PCR）技术可能是有价值的。PCR技术允许扩增肿瘤特异性DNA序列或mRNA序列（逆转录成cDNA后），如果侧翼序列是明确定义的。这种PCR介导的扩增可以检测来自许多正常细胞之间的仅少数恶性细胞的特定序列。明确定义的染色体易位已被用作肿瘤特异性标记，如t（9;22）。使用特异性染色体畸变作为肿瘤特异性标志物的优点是它们在疾病过程中的稳定性。然而，只有10-15%的ALL和25-30%的AML具有特定的染色体易位，并且其中很大一部分的精确断点尚不清楚。近年来的研究表明，以可变（V）和连接（J）基因特异性寡核苷酸为引物，通过PCR介导的重排免疫球蛋白（IG）和T细胞受体（TcR）基因连接区扩增，可以检测急性白血病的MRD。该应用的主要缺陷是在诊断时发生多个重排（寡克隆性）和在复发时重排模式的变化（克隆进化），这将导致该MRD-PCR技术的假阴性结果。总之，检测MRD的技术选择取决于白血病的免疫表型、明确的染色体易位的存在、重排的IG和/或TcR基因的存在以及免疫表型转变和IG和TcR基因重排模式变化的可能性。（400字处截断摘要）
Detection of minimal residual disease (MRD) can be useful for adaptation or stratification of treatment in acute leukemia patients and may finally result in individualization of treatment protocols. Although leukemic cells generally have immunophenotypes comparable to their normal counterparts, it is possible to use immunological marker analysis for the detection of MRD based on the assumption that the presence of positive cells outside their normal breeding sites and 'homing areas' is indicative of malignancy. This approach can be used for the detection of MRD in blood and bone marrow of patients with a terminal deoxynucleotidyl transferase (TdT) positive T-cell acute lymphoblastic leukemia (ALL) and patients with a TdT+ acute myeloid leukemia (AML) as well as in cerebrospinal fluid of patients with a TdT+ leukemia. In other types of acute leukemias, immunological marker analysis generally does not allow detection of low frequencies of malignant cells, but in a part of them the polymerase chain reaction (PCR) technique may be valuable. The PCR technique allows the amplification of tumor-specific DNA sequences or mRNA sequences (after reverse transcription into cDNA), if the flanking sequences are well-defined. This PCR-mediated amplification can detect specific sequences which are derived from only a few malignant cells between many normal cells. Well-defined chromosome translocations have been used as tumor-specific markers, such as t(9;22). An advantage of using specific chromosome aberrations as tumor-specific markers is their stability during the disease course. However, only 10-15% of ALL and 25-30% of AML have a specific chromosome translocation and in a large part of them the precise breakpoints are not (yet) known. Recent studies indicate that it is possible to detect MRD in acute leukemias by use of PCR-mediated amplification of the junctional regions of rearranged immunoglobulin (Ig) and T-cell receptor (TcR) genes, using variable (V) and joining (J) gene-specific oligonucleotides as primers. Major pitfalls of this application are the occurrence of multiple rearrangements at diagnosis (oligoclonality) and changes in rearrangement patterns at relapse (clonal evolution), which will lead to false negative results of this MRD-PCR technique. In conclusion, the technique of choice for the detection of MRD is dependent on the immunophenotype of the leukemia, the presence of a well-defined chromosome translocation and the presence of a rearranged Ig and/or TcR gene as well as the chance of immunophenotypic shifts and changes in Ig and TcR gene rearrangement patterns.(ABSTRACT TRUNCATED AT 400 WORDS)