影响恶性肿瘤患者造血系统自体外周血造血干细胞数量和质量的因素分析(英文)

Effective factors for quantity and quality of auto-peripheral blood stem cells in hematopoietic system of patients with malignant tumor

背景:造血系统恶性肿瘤的造血重建除与疾病本身、预处理方案、移植后支持治疗手段等相关外,自体外周血造血干细胞的动员、采集和冻存是影响其移植后造血系统顺利重建的关键因素。目的:观察造血系统恶性肿瘤患者自体外周血造血干细胞经动员、采集和冻存后,重新回输至造血系统的重建情况,并分析影响外周血造血干细胞数量和质量的因素。设计:以造血系统恶性肿瘤为对象的病例分析。单位:解放军广州军区广州总医院血液科,南方医科大学珠江医院血液科。对象:选取2000-02/2004-12解放军广州军区广州总医院血液科收治的18例造血系统恶性肿瘤住院患者,年龄16～56岁,其中急性髓性白血病2例,急性淋巴细胞白血病1例,淋巴瘤白血病2例,慢性粒细胞白血病2例,多发性骨髓瘤4例,非霍奇金淋巴瘤7例。粒细胞集落刺激因子(Granocyte,中外制药产品,批号N3G31)。方法:①全部病例均采用对肿瘤有效的联合化疗方案+粒细胞集落刺激因子进行动员。联合化疗方案:白血病患者第1～3天每隔12h给予阿糖胞苷2g/m2,第1～5天给予足叶乙甙200mg/m2或氟达拉宾50mg/m2。多发性骨髓瘤患者给予阿糖胞苷方案同上,第1～2天给予环磷酰胺1g/m2。淋巴瘤患者第1～2天给予环磷酰胺2g/m2。各类型患者化疗后白细胞降至1.0×109L-1以下时开始进行粒细胞集落刺激因子动员,5μg/(kg.d)皮下注射至采集结束。②当白细胞恢复至(4.0～10.0)×109L-1时开始采集外周血造血干细胞,单个核细胞计数≥4.0×108/kg或CD34+细胞≥2.0×106/kg时结束采集,经程序降温仪处理置入-196℃液氮中保存,37～40℃水浴解冻。③患者病灶部位行局部照射预处理,200cGy/次,5次/周,连续4周,总剂量40Gy。结束后48h回输外周血造血干细胞(55.3±28.7)mL,回输日距采集日平均为(56.5±22.3)d。全部患者于干细胞移植后第1天起皮下注射粒细胞集落刺激因子300μg/d,至中性粒细胞≥0.5×109L-1时停止。检测冻存前及解冻后自体外周血造血干细胞的锥虫蓝拒染率、单个核细胞计数、粒-单系祖细胞集落数及CD34+细胞百分率。主要观察指标:①自体外周血造血干细胞的采集情况。②冻存后自体外周血造血干细胞存活率及相关指标检测。③自体外周血造血干细胞移植后造血系统重建情况。结果:18例造血系统恶性肿瘤患者全部进入结果分析。①18例患者自体外周血造血干细胞平均采集时间为化疗后12.6d,采集次数为1.9次,采集第1天白细胞总数为(8.93±1.27)×109L-1,单个核细胞采集率为(138.33±28.61)%。②冻存后18例自体外周血造血干细胞标本锥虫蓝拒染率与冻存前基本相似[(96.26±1.33)%,(92.75±2.04)%,P>0.05]。解冻后单个核细胞、CD34+、粒-单系祖细胞回收率分别为(91.96±1.37)%,(85.94±0.64)%,(87.69±4.53)%。骨髓瘤患者的单个核细胞采集率、CD34+细胞百分率及粒-单系祖细胞集落数均明显低于白血病和淋巴瘤患者(t=2.524～3.268,P<0.05)。③移植后15d,15例患者中性粒细胞恢复至≥0.5×109L-1;移植后20d,血小板恢复至≥20×109L-1。化疗疗程>10次的5例患者粒-单系祖细胞生长不良,为(18.67～26.82)×105/kg,其中3例出现自体外周血造血干细胞移植后造血重建延迟。结论:①重组粒细胞集落刺激因子与大剂量化疗联合的动员方案可缩短外周血造血干细胞采集时间,提高单个核细胞采集率。②移植前化疗次数增多可影响自体外周血造血干细胞的数量和质量,导致造血重建延迟。

BACKGROUND： Hematopoietic reconstruction of malignant tumor in hematopoietic system is related to disease itself, pretreatment program and therapeutic tool after transplantation; especially, mobilization, collection and cryopreservation of auto-peripheral blood stem cell play a key role in successful reconstruction of hematopoietic system after transplantation. OBJECTIVE： To investigate the reconstruction of hematopoietic system through mobilization, collection and cryopreservation of auto-peripheral blood stem cell in patients with malignant tumor and analyze the effective factors on quantity and quality of auto-peripheral blood stem cell. DESIGN ： Case analysis based on malignant tumor in hematopoietic system SETTING： Department of Blood, Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA; Department of Blood, Zhujiang Hospital, Nanfang Medical University. PARTICIPANTS： A total of 18 patients with malignant tumor in hematopoietic system were selected from Department of Blood, Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA. Their ages ranged from 16 to 56 years. Among them, 2 patients had acute myelogenous leukemia （AML）, 1 acute lymphoblastic leukemia （ALL）, 2 lymphoblastic leukemia （LL）, 2 chronic granulocytic leukemia （CGL）, 4 multiple myeloma （MM）, and 7 non-Hodgkin lymphoma. Granulocyte colony-stimulating factor （G-CSF） was made by Chugai Pharmaceutical Company Limited （batch number： N3G31 ）. METHODS： （1） All patients were mobilized with associated chemotherapy ＋ G-CSF. Associate chemotherapy： Patients with leukemia were given 2 g/m^2 arabinosyl cytosine every 12 hours from the first to the third days and 200 mg/m^2 etoposide or 50 mg/m^2 fludarabine from the first to the fifth days. In addition, patients with MM were treated with arabinosyl cytosine as the same way mentioned above and with 1 g/m^2 cyclophosphamide from the first to the second days. And patients with lymphoma were given 2 g/m^2 cyclophosphamide from the first to the second days. When numbers of leucocyte of all patients decreased below 1.0×10^9 L^-1 after chemotherapy, G-CSF started mobilization and the collection was stopped with 5 μg/（kg .d） subcutaneous injection. （2）When numbers of leucocyte increased to （4.0-10.0）×10^9 L^-1, hemopoietic stem cells of peripheral blood were collected till the amount of mononuclear cells ≥ 4.0×10^8/kg or numbers of CD34^＋ cells ≥ 2.0×10^6/kg. And then, the samples were dealt with cooling device, maintained in liquid nitrogen at -196 % and defrosted in water bath at 37-40 %. （3） Focal sites of patients were pretreated with local irradiation with 200 cGy/time and 5 times/week for 4 successive weeks. The total dosage was 40 Gy. At 48 hours later, （55.3±28.7） mL hemopoietic stem cells of peripheral blood were transfused back, and the duration from transfusion to collection was about （56.5±22.3） days. 300μg/d G-CSF was subcutaneously injected into all patients at 1 day after transplantation and the reaction was stopped at the phase of neutrophil ≥ 0.5×10^9 L^-1. Finally, refusing-staining rate of trypan blue of peripheral blood stem cell, amount of mononuclear cells, number of granulation-monophyly progenitor cell colony and percentage of CD34^＋ cells were detected before and after thaw.MAIN OUTCOME MEASURES： （1） Collection of auto-peripheral blood stem cell; （2） survival rate and related markers of auto-peripheral blood stem cell after cryopreservation; （3） hematopoietic reconstruction of auto-peripheral blood stem cell after transplantation. RESULTS： All 18 patients with malignant tumor in hematopoietic system were involved in the final analysis. The mean collection time of auto-peripheral blood stem cell was 12.6 days after chemotherapy, the collection times were 1.9, total number of leucocyte was （8.93±1.27） × 10^9 L^-1 on the first day, and collection rate of mononuclear cell was （138.33± 28.61）%. （2） Refusing-staining rate of trypan blue of auto-peripheral blood stem cell was similar before and after cryopreservation [（96.26±1.33）%, （92.75±2.04）%, P〉 0.05]. In addition, after cryopreservation, recovery rates of mononuclear cells, CD34^＋ cells and granulation-monophyly progenitor cell were （91.96±1.37）%, （85.94±0.64）% and （87.69±4.53）%, respectively. Collection rate of mononuclear cells, number of granulation-monophyly progenitor cell colony and percentage of CD34^＋ cells were lower in patients with myeloma than in those with leukemia and lymphoma （t = 2.524-3.268, P〈 0.05）. （3） At 15 days after transplantation, 15 patients had the neutrophil ≥ 0.5 × 10^9 L^-1; at 20 days after transplantation, blood platelet was ≥ 20 × 10^9 L^-1 granulation-monophyly progenitor cells [（18.67-26.82） × 10^5/kg] of 5 patients grew poorly if the course of chemotherapy was more than 10 times. Among them, 3 patients had delayed hematopoietic reconstruction after transplantation of auto-peripheral blood stem cell. CONCLUSION： （1） High-dose chemotherapy combined with G-CSF can shorten collection time of peripheral blood stem cell and improve collection rate of mononuclear cells. （2） Increase of chemotherapy times before transplantation can affect quantity and quality of auto-peripheral blood stem cell and cause delayed hematopoietic reconstruction.