Nagafuji K, Harada M, Takamatsu Y, Eto T, Teshima T, Kamura T, Okamura T, Hayashi S, Akashi K, Murakawa M
First Department of Internal Medicine, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Br J Haematol. 1993 Nov;85(3):578-83. doi: 10.1111/j.1365-2141.1993.tb03351.x.
A major issue in autologous blood stem cell transplantation (ABSCT) for leukaemia is whether peripheral blood stem cell (PBSC) harvests are less contaminated with leukaemic cells than bone marrow mononuclear cells (BMMNC). We compared leukaemic contamination in PBSC harvests and BMMNC, obtained simultaneously, by using reverse transcriptase polymerase chain reaction (RT-PCR) of leukaemia-specific chimaeric messenger RNA (mRNA), in three patients with Philadelphia chromosome (Ph)-positive acute lymphoblastic leukaemia (ALL), one with Ph-positive acute myelogenous leukaemia (AML), and two with acute promyelocytic leukaemia (APL). Our two-step PCR method employed 'nested primers' in the second step and can detect one leukaemic blast diluted into 10(6) HL-60 cells. In three of four patients with Ph-positive ALL and AML we detected leukaemic contamination in both PBSC harvests and BMMNC. In the remaining patient with ALL, both PBSC harvests and BMMNC were PCR-negative. Both PBSC harvests and BMMNC from one patient with APL were PCR-positive. In contrast, PBSC harvests from another patient with APL, whose BMMNC could not be obtained because of bone marrow necrosis, were PCR-positive after the first course of consolidation chemotherapy, but became PCR-negative after the second course. The present study does not support the hypothesis that PBSC harvests are less contaminated by leukaemic cells than BMMNC, but suggests that PBSC harvests are contaminated when BMMNC are contaminated.
白血病自体血干细胞移植（ABSCT）中的一个主要问题是，外周血干细胞（PBSC）采集物中白血病细胞的污染是否低于骨髓单个核细胞（BMMNC）。我们对3例费城染色体（Ph）阳性急性淋巴细胞白血病（ALL）患者、1例Ph阳性急性髓性白血病（AML）患者和2例急性早幼粒细胞白血病（APL）患者同时采集的PBSC采集物和BMMNC中的白血病污染情况进行了比较，采用白血病特异性嵌合信使核糖核酸（mRNA）的逆转录聚合酶链反应（RT-PCR）技术。我们的两步PCR方法在第二步使用了“巢式引物”，能够检测出稀释到10(6)个HL-60细胞中的一个白血病母细胞。在4例Ph阳性ALL和AML患者中，有3例在PBSC采集物和BMMNC中均检测到白血病污染。在其余1例ALL患者中，PBSC采集物和BMMNC的PCR检测均为阴性。1例APL患者的PBSC采集物和BMMNC的PCR检测均为阳性。相比之下，另1例APL患者因骨髓坏死无法获得BMMNC，其PBSC采集物在巩固化疗的第一个疗程后PCR检测为阳性，但在第二个疗程后变为PCR阴性。本研究不支持PBSC采集物中白血病细胞污染低于BMMNC的假说，但提示当BMMNC受到污染时，PBSC采集物也会受到污染。
