Kletzel M, Longino R, Rademaker A W, Danner-Koptik K E, Olszewski M, Morgan E R
Department of Pediatrics, The Children's Memorial Hospital, Northwestern University School of Medicine, Chicago, Illinois 60614, USA.
Pediatr Transplant. 1998 Aug;2(3):191-6.
The purpose of this study was to determine the feasibility and assess optimal timing of harvesting peripheral blood stem cells (PBSC) for transplantation in young children. Thirteen children with body weight less than 25 kg, mean age of 3.9 years (1-9 yrs) who had recurrent solid tumors and leukemia were given tumor specific chemotherapy followed by i.v. rhG-CSF (5 microg/kg/d) for stem cell mobilization. Cytaphereses were done through a central venous line (CVL) during the marrow recovery phase (WBC >0.5 x 10(9)/l). The phereses were analyzed separately and assigned to three groups depending on the WBC at the time of the pheresis: Group I (WBC <1.0 x 10(9)/l), Group II [WBC in the range 1.0-3.0 x 10(9)/l] and Group III (WBC >3.0 x 10(9)/l). Samples from each harvest were assayed for cell count, CFU-GM, BFU-E, CD34+ cell count, and tumor cell immunocytology in patients with neuroblastoma (NBL). A median of 3.2 x 10(8) mononuclear cells per kg (MNC/kg), [mean 2.8 x 10(8) MNC/kg, standard error of the mean (SEM) +/- 0.74 (1.1-4.7)] were infused following myeloablative therapy. 78 phereses were performed in 13 children with a median weight of 18 kg (10-25 kg). A median of 5 phereses were performed per patient. There were no significant differences in the percentage and number of CD34+ cells, CFU-GM or BFU-E colonies assayed by plating 0.5 x 10(5) cells. Differences could be found in the total number of MNC (p<0.008) and the number of MNC/kg (p<0.001) between Groups II and III. No tumor cell contamination was detected in the NBL patients by immunocytology. All patients were rescued with PBSC and achieved sustained white cell engraftment (ANC >0.5 x 10(9)/l) at a median of 13.5 d (10-25 d) and platelet engraftment (untransfused platelet count >20.0 x 10(9)/l) at a median of 29 d (12-63 d). The only toxicity encountered during the phereses was thrombocytopenia in 4 patients whose median post-pheresis platelet count was 6.0 x 10(9)/l (3.0-9.01). It is concluded that collection of PBSC in young children is feasible and safe and can be performed through a cuffed CVL at the time of WBC recovery post mobilization with chemotherapy and G-CSF. Cytopheresis can be effectively performed when the peripheral WBC count approaches 1.0 x 10(9)/l. Following stem cell infusion, engraftment was prompt and durable.
本研究的目的是确定在幼儿中采集外周血干细胞（PBSC）用于移植的可行性，并评估最佳采集时机。13名体重小于25kg、平均年龄3.9岁（1 - 9岁）的复发性实体瘤和白血病患儿接受了肿瘤特异性化疗，随后静脉注射重组人粒细胞集落刺激因子（rhG - CSF，5μg/kg/d）以动员干细胞。在骨髓恢复阶段（白细胞>0.5×10⁹/L）通过中心静脉导管（CVL）进行血细胞分离术。根据血细胞分离术时的白细胞水平将采集物分别分析并分为三组：第一组（白细胞<1.0×10⁹/L）、第二组[白细胞在1.0 - 3.0×10⁹/L范围内]和第三组（白细胞>3.0×10⁹/L）。对每位患儿每次采集的样本进行细胞计数、粒 - 巨噬细胞集落形成单位（CFU - GM）、爆式红系集落形成单位（BFU - E）、CD34⁺细胞计数检测，对神经母细胞瘤（NBL）患者的样本进行肿瘤细胞免疫细胞学检测。清髓性治疗后，每千克体重输注的单个核细胞（MNC）中位数为3.2×10⁸个/kg（平均2.8×10⁸个/kg，平均标准误（SEM）±0.74（1.1 - 4.7））。对13名中位体重18kg（10 - 25kg）的患儿进行了78次血细胞分离术。每位患者血细胞分离术的中位数为5次。通过接种0.5×10⁵个细胞检测，CD34⁺细胞、CFU - GM或BFU - E集落的百分比和数量无显著差异。第二组和第三组之间的单个核细胞总数（p<0.008）和每千克体重单个核细胞数（p<0.001）存在差异。通过免疫细胞学检测，未在NBL患者中检测到肿瘤细胞污染。所有患者均接受PBSC救治，白细胞中位植入时间为13.5天（10 - 25天）（中性粒细胞绝对值（ANC）>0.5×10⁹/L），血小板中位植入时间为29天（12 - 63天）（未输血时血小板计数>20.0×10⁹/L）。血细胞分离术期间仅4例患者出现血小板减少这一毒性反应，其血细胞分离术后血小板计数中位数为6.0×10⁹/L（3.0 - 9.01）。结论是，在幼儿中采集PBSC是可行且安全的，可在化疗和G - CSF动员后白细胞恢复时通过带袖套的CVL进行。当外周白细胞计数接近1.0×10⁹/L时可有效进行血细胞分离术。干细胞输注后，植入迅速且持久。
