Mariette X, Fermand J P, Brouet J C
Laboratory of Immunopathology, Hôpital Saint-Louis, Paris, France.
Bone Marrow Transplant. 1994 Jul;14(1):47-50.
In order to assess the contamination with malignant cells of peripheral blood stem cell (PBSC) transplants used to support high-dose therapy in multiple myeloma (MM), we used the immunoglobulin heavy chain gene radioactive fingerprinting polymerase chain reaction (PCR) method to detect clonal cells in PBSC from 10 patients. The sensitivity of the technique allowed the detection of one clonal cell among 10(4) normal blood mononuclear cells. A clonal band was detected in 4 of 11 leukaphereses samples. The level of contamination was low because a clonal band could never be identified on ethidium bromide-stained agarose gels whose sensitivity is between 1 and 5%. The use of granulocyte-colony stimulatory factor (G-CSF) in combination with chemotherapy in three cases did not seem to increase the contamination of PBSC grafts; in one patient, G-CSF was used during a second course of leukapheresis which was free of detectable clonal cells whereas the first one performed after chemotherapy alone contained clonal cells. Thus, PBSC grafts may rarely be completely devoid of clonal potentially malignant cells but the level of contamination is much lower than in BM grafts. Whether graft contamination is an important adverse prognostic factor for patients with MM undergoing intensive treatment and autografting is still unsettled.
为了评估用于支持多发性骨髓瘤（MM）高剂量治疗的外周血干细胞（PBSC）移植中恶性细胞的污染情况，我们采用免疫球蛋白重链基因放射性指纹聚合酶链反应（PCR）方法检测了10例患者PBSC中的克隆细胞。该技术的灵敏度能够检测出10⁴个正常血液单核细胞中的一个克隆细胞。在11个白细胞分离样本中的4个检测到了克隆条带。污染水平较低，因为在灵敏度为1%至5%的溴化乙锭染色琼脂糖凝胶上从未鉴定出克隆条带。在3例患者中，粒细胞集落刺激因子（G-CSF）与化疗联合使用似乎并未增加PBSC移植物的污染；在1例患者中，第二次白细胞分离过程中使用了G-CSF，未检测到克隆细胞，而仅在化疗后进行的第一次白细胞分离中含有克隆细胞。因此，PBSC移植物可能很少完全没有克隆性潜在恶性细胞，但污染水平远低于骨髓移植物。对于接受强化治疗和自体移植的MM患者，移植物污染是否是一个重要的不良预后因素仍未确定。
