Leung W, Chen A R, Klann R C, Moss T J, Davis J M, Noga S J, Cohen K J, Friedman A D, Small D, Schwartz C L, Borowitz M J, Wharam M D, Paidas C N, Long C A, Karandish S, McMannis J D, Kastan M B, Civin C I
Johns Hopkins Oncology Center, Baltimore MD, USA.
Bone Marrow Transplant. 1998 Nov;22(10):971-9. doi: 10.1038/sj.bmt.1701471.
Many poor-risk neuroblastomas and tumours of the Ewing's sarcoma family (ET) recur despite autologous transplants. Recurrence may be due to tumor cells contained in the BM harvests or PBSC harvests. The objectives of this prospective study were to: (1) determine the incidence and degree of tumor cell contamination in paired BM and PBSC harvests; and (2) determine the efficacy of tumor cell purging by immunomagnetic CD34+ cell selection. 198 samples from 11 consecutive patients with neuroblastoma or Ewing's sarcoma were analyzed. We assayed tumor contamination by RT-PCR assay for PGP 9.5, plus immunohistochemistry for neuroblastoma-specific antigens (the latter in neuroblastoma only). None of these patients had tumor cells detected in their BM by clinical histology immediately before BM or PBSC harvests. However, 82% of PBSC and 89% of backup BM harvests were contaminated with tumor by RT-PCR and/or immunocytochemistry assays. Unselected PBSC and BM harvests contained similar quantities of tumor cells (median, approximately 200000 cells). Cyclophosphamide plus G-CSF mobilization did not affect the incidence or level of contamination in PBSC harvests, as compared to blood obtained before mobilization. Immunomagnetic CD34+ cell selection depleted tumor cells by a median of 3.0 logs for PBSC, and 2.6 logs for BM harvests.
许多高危神经母细胞瘤和尤因肉瘤家族肿瘤（ET）即便进行了自体移植仍会复发。复发可能是由于骨髓采集物或外周血干细胞采集物中所含的肿瘤细胞。这项前瞻性研究的目的是：（1）确定配对的骨髓和外周血干细胞采集物中肿瘤细胞污染的发生率和程度；（2）确定通过免疫磁珠法选择CD34+细胞清除肿瘤细胞的疗效。对11例连续的神经母细胞瘤或尤因肉瘤患者的198份样本进行了分析。我们通过针对PGP 9.5的逆转录聚合酶链反应（RT-PCR）检测以及神经母细胞瘤特异性抗原的免疫组织化学检测（后者仅用于神经母细胞瘤）来测定肿瘤污染情况。在进行骨髓或外周血干细胞采集前，通过临床组织学检查，这些患者的骨髓中均未检测到肿瘤细胞。然而，通过RT-PCR和/或免疫细胞化学检测，82%的外周血干细胞采集物和89%的备用骨髓采集物被肿瘤污染。未经选择的外周血干细胞和骨髓采集物中所含肿瘤细胞数量相似（中位数约为200000个细胞）。与动员前采集的血液相比，环磷酰胺加粒细胞集落刺激因子动员并未影响外周血干细胞采集物中污染的发生率或水平。免疫磁珠法选择CD34+细胞对外周血干细胞采集物中的肿瘤细胞清除中位数为3.0个对数级，对骨髓采集物为2.6个对数级。
