Faulkner L B, Tintori V, Tamburini A, Paoli A, Garaventa A, Viscardi E, Tucci F, Lippi A A, De Bernardi B, Bernini G
Department of Pediatrics, University of Florence, Ospedale Pediatrico A. Meyer, Italy.
J Hematother. 1998 Aug;7(4):361-6. doi: 10.1089/scd.1.1998.7.361.
High-sensitivity immunocytochemistry was used to evaluate the relative frequency of neuroblastoma cells in bone marrow and peripheral blood in patients with neuroblastoma (NB). A total of 51 concomitant paired blood and marrow samples (102 total) from 35 patients with NB (age 4 months-31 years; stage 29 stage IV, 4 stage III, 2 stage IVS; 14 at diagnosis, 18 in relapse, 12 during treatment, and 7 off-therapy) were analyzed. Cytospins containing up to 10(6) cells each were prepared using the mononuclear cell (MNC) fraction. For immunocytologic staining, a primary mouse monoclonal anti-GD2 antibody (3F8), a secondary antimouse biotinylated antibody, and a streptavidin-alkaline phosphatase complex were used. A minimum of two cytospins containing a mean of 1.4 x 10(6) total MNCs was analyzed in addition to a negative and a positive control. No circulating tumor cells were detected when the concomitant marrow samples were negative or had <10 positive cells per 106 MNC (23 of 51 samples). Of the 18 marrow samples positive at 10-10,000 cells per 106 MNC, 6 had detectable NB cells in the corresponding blood sample, whereas for marrow samples with >10,000 NB cells per 10(6) MNC (1%), the concomitant blood sample was positive for 9 of the 10. When both marrow and blood samples were positive (15 BM-PB pairs), NB cell frequency was significantly lower in blood, with a mean difference of 2.14 logs (median 2.22, range -0.16-4.8, standard error 0.38). In patients with NB, circulating tumor cell frequencies seem to be substantially lower than in concomitant marrow samples, with a mean difference of >2 logs.
采用高灵敏度免疫细胞化学方法评估神经母细胞瘤（NB）患者骨髓和外周血中神经母细胞瘤细胞的相对频率。分析了来自35例NB患者（年龄4个月至31岁；29例IV期、4例III期、2例IV-S期；14例诊断时、18例复发时、12例治疗期间、7例治疗结束后）的总共51对同时采集的血液和骨髓样本（共102份）。使用单核细胞（MNC）组分制备每个含有多达10⁶个细胞的细胞涂片。免疫细胞化学染色时，使用小鼠抗GD2单克隆抗体（3F8）、抗小鼠生物素化二抗和链霉亲和素-碱性磷酸酶复合物。除阴性和阳性对照外，至少分析两个平均含有1.4×10⁶个总MNC的细胞涂片。当同时采集的骨髓样本为阴性或每10⁶个MNC中阳性细胞少于10个时（51份样本中的23份），未检测到循环肿瘤细胞。在每10⁶个MNC中含有10至10000个细胞的18份骨髓样本中，6份在相应血液样本中可检测到NB细胞，而对于每10⁶个MNC中NB细胞>10000个（1%）的骨髓样本，10份中有9份相应血液样本呈阳性。当骨髓和血液样本均为阳性时（15对BM-PB），血液中NB细胞频率显著较低，平均差异为2.14个对数（中位数2.22，范围-0.16至4.8，标准误0.38）。在NB患者中，循环肿瘤细胞频率似乎显著低于同时采集的骨髓样本，平均差异>2个对数。
