Miyamoto T, Nagafuji K, Harada M, Eto T, Fujisaki T, Kubota A, Akashi K, Mizuno S, Takenaka K, Kanaji T
First Department of Internal Medicine, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Br J Haematol. 1995 Sep;91(1):132-8. doi: 10.1111/j.1365-2141.1995.tb05258.x.
Peripheral blood stem cells (PBSC) have been used increasingly for haemopoietic reconstitution after marrow-ablative chemotherapy in patients with acute leukaemia because of the possibility that there is a lower risk of leukaemic contamination. We have developed a titration assay using a competitive reverse transcriptase polymerase chain reaction (RT-PCR) which is able to estimate the number of AML1/ETO transcripts so that minimal residual disease (MRD) can be monitored quantitatively in patients with t(8;21) acute myelogenous leukaemia (AML). Using a qualitative RT-PCR method, AML1/ETO transcripts could be detected in all samples from 15 first PBSC harvests and 11 second PBSC harvests obtained from 15 patients with t(8;21) AML. With our competitive RT-PCR assay, the number of AML1/ETO transcripts was found to be lower in the second PBSC harvest than that in the first in every individual. Furthermore, MRD in PBSC harvests was less than that in the corresponding bone marrow obtained on the day of PBSC collection in the individual patients studied. In 10 patients who received autologous blood stem cell transplantation (ABSCT), we could not find a relationship between the number of AML1/ETO transcripts in the infused PBSC harvests and the clinical outcome after ABSCT. The present study clearly indicates that although PBSC harvests collected after consolidation chemotherapy are contaminated by leukaemic cells, the degree of leukaemic contamination may decrease as chemotherapy is repeated. The mobilization of PBSC by repeated chemotherapy may provide an advantageous source of haemopoietic stem cells for ABSCT.
外周血干细胞（PBSC）越来越多地用于急性白血病患者进行骨髓清除性化疗后的造血重建，因为其白血病污染风险可能较低。我们开发了一种滴定分析法，使用竞争性逆转录聚合酶链反应（RT-PCR）来估计AML1/ETO转录本的数量，从而能够对t(8;21)急性髓性白血病（AML）患者的微小残留病（MRD）进行定量监测。使用定性RT-PCR方法，在15例t(8;21) AML患者的15次首次PBSC采集样本和11次第二次PBSC采集样本中均能检测到AML1/ETO转录本。通过我们的竞争性RT-PCR分析，发现每个个体第二次PBSC采集时AML1/ETO转录本的数量均低于首次采集时。此外，在所研究的个体患者中，PBSC采集中的MRD低于PBSC采集当天相应骨髓中的MRD。在10例接受自体血干细胞移植（ABSCT）的患者中，我们未发现输注的PBSC采集中AML1/ETO转录本数量与ABSCT后的临床结局之间存在关联。本研究清楚地表明，尽管巩固化疗后采集的PBSC受到白血病细胞污染，但随着化疗的重复，白血病污染程度可能会降低。重复化疗动员PBSC可能为ABSCT提供有利的造血干细胞来源。
