多西他赛联合重组人粒细胞集落刺激因子动员乳腺癌患者外周血干细胞的效果及影响因素分析

 目的研究多西他赛联合重组人粒细胞集落刺激因子（rhG-CSF）在乳腺癌患者动员外周血干细胞 （peripheral blood stem cell,PBSC）的效果及影响因素。方法2006年2月—2009年6月我科收治的55例 行多西他赛（商品名多帕菲,TXT,120 mg/m2 持续静脉滴注3 h）动员的乳腺癌患者,化疗后白细胞降至 1.0×109/L左右时使用rhG-CSF 5 μg /(kg·d) 动员至采集结束。并进一步分析患者年龄、化疗后白细 胞最低数、采集前各类血细胞数、内脏、骨转移情况及化、放疗情况对采集单个核细胞（mononuclear cell,MNC）和CD34+细胞的影响。结果多西他赛化疗动员后中位7d白细胞降至1.0×109/L 左右,皮下注 射rhG-CSF中位3d,采集总MNC平均(5.51±1.24)×108/kg,采集总CD34+细胞平均(2.90±1.38)×106/kg 。年龄、化疗后白细胞最低数与采集CD34+细胞均显著相关（P<0.05）。化疗的周期数≤6的患者采集平均 CD34+细胞数较周期数≥6患者显著增高（P<0.05）。无1例患者出现严重不良反应。结论TXT（120 mg/m2 持续静脉滴注3 h）联合rhG-CSF为转移性乳腺癌患者动员的有效安全方案。患者年龄、既往化疗周期数显 著影响患者CD34+细胞的采集数量。患者化疗后白细胞最低数可作为采集CD34+细胞数量的预测指标。     

供者外周血干细胞动员采集198例临床分析

 目的　研究外周血造血干细胞移植（PBSCT）供者外周血干细胞动员采集的方法及其效果。方法　198名健康供者每天皮下注射重组人粒细胞集落刺激因子（rhG-CSF）（5～10）μg／kg进行外周血干细胞动员,第5天开始采集。采用血细胞分析仪行单个核细胞（MNC）计数,流式细胞术（FCM）行CD+34 细胞计数。分析供者性别、身高、年龄、采集当天外周血白细胞（WBC）计数对动员采集效果的影响。结果　所有供者均成功动员采集,采集当天的MNC计数平均为（4.19±1.96）×108／kg,CD+34 细胞计数平均为（2.98±1.40）×106／kg;MNC和CD+34细胞计数与供者性别、身高、年龄无关;采集当天外周血WBC计数与MNC、CD+34 细胞计数呈正相关（r＝0.9201,P＝0.0035;r＝0.8420,P＝0.0149）;采集当天外周血WBC计数≥20.0×109／L的供者比＜20.0×109／L的供者采集效果更显著（F＝4.688,P＝0.0013;F＝4.622,P＝0.0006）。结论　rhG-CSF动员的健康供者采集当天外周血WBC计数是一项预测CD+34细胞采集数量的简单、可行的指标。     

外周血CD_(34)~+细胞检测对外周血干细胞采集结果及时机选择的意义

 目的　研究外周血CD+34 细胞数对采集结果的意义,并探索可用于临床指导外周干细胞采集时机选择的参考阈值。方法　2007年1月至2009年12月共57例次自体造血干细胞移植动员采集患者,以环磷酰胺（CTX）化疗+粒细胞集落刺激因子（G-CSF）（5～10 μg／kg）动员,COBE分离仪（Spectra Version 6）行外周血造血干细胞采集,应用流式细胞术监测外周血中CD+34 细胞绝对计数。结果　采集产品单个核细胞（MNC）中位数4.6×108／kg（0.3×108／kg～10.5×108／kg）,CD+34 细胞中位数2.4×106／kg（0.16×106／kg～34.9×106／kg）,外周血CD+34 细胞数是产品MNC和CD+34 细胞总量唯一相关指标,外周血白细胞（WBC）与采集产品MNC和CD+34 细胞数无关。进一步分析提示外周血CD+34 计数≥15／μl,单次采集效率提高,CD+34 细胞采集量达1×106／kg和2×106／kg比例为81 %和60 %,采集产品MNC和CD+34 总数明显提高。提示外周血CD+34 细胞数15／μl可作为启动采集。ROC分析发现外周血CD+34 细胞 25（26.5～28.6）／μl,单次采集足量CD+34 细胞概率最大。结论　外周血CD+34 细胞计数是外周血自体干细胞采集重要的相关指标,CD+34 细胞 15／μl可作为采集时机选择的阈值。     

环磷酰胺联合G-CSF动员慢性粒细胞白血病患者外周血造血干细胞

 目的 评价环磷酰胺（Cy）联合粒细胞集落刺激因子（G-CSF）动员慢性粒细胞白血病（CML）患者外周血造血干细胞的动员、采集效果及不良反应。方法 选择符合诊断的CML患者7例,无心、肺、肝、肾等重要脏器功能损害。动员方案为Cy+人基因重组粒细胞集落刺激因子（rhG-CSF）,于化疗后WBC降至1.0×109／L时开始用rhG-CSF,用CS3000 plus血细胞分离机采集干细胞,-80 ℃低温冰箱冻存,并作单个核细胞（MNC）计数及CD+34细胞测定。结果 Cy平均剂量为4.0（3.0 ~ 5.0）g,rhG-CSF的剂量均为300 μg／d,平均5.8（3 ~ 9）d;其中仅1例患者动员2次,余6例均动员1次;共采集2次,每次循环血量为10 ~ 13 L;采集的MNC达（2.94 ~ 5.45）×108／kg,CD+34细胞数达（2.15 ~ 6.59）×106／kg ;患者能耐受整个动员和采集过程,无一例因化疗的不良反应而中止动员;移植后全部患者均迅速恢复了造血功能。结论 Cy联合G-CSF的动员方案对CML患者是安全、有效的。     

应用粒细胞集落刺激因子动员后异基因外周血干细胞移植供者外周血干细胞与不成熟粒细胞的相关性

 目的　探讨供者应用重组人粒细胞集落刺激因子（rhG-CSF）动员后单采外周血干细胞（PBSC）移植物中的不成熟粒细胞与CD+34 细胞及单个核细胞（MNC）之间的相关性。方法　122例异基因外周血干细胞移植（allo-PBSCT）供者用rhG-CSF 7.25～10 μg·kg-1·d-1进行动员,于动员前及动员后对外周血及PBSC收集物中的MNC和不成熟粒细胞及CD+34 细胞进行检测,并计算出输给患者的MNC、不成熟粒细胞及CD+34 细胞数。结果　动员后白细胞计数及不成熟粒细胞比值逐渐升高,至第5天达高峰;外周血MNC中不成熟粒细胞与CD+34 细胞同步增加有较好的相关性;采集后的PBSC移植物输给患者后,均全部植活并恢复造血功能,为染色体核型或血型及HLA转为供者型所证实,慢性粒细胞白血病患者Ph1染色体转阴。结论　rhG-CSF 7.25～10 μg·kg-1·d-1分两次皮下注射能有效动员PBSC;allo-PBSCT供者经rhG-CSF动员后,外周血MNC中不成熟粒细胞与CD+34细胞数同步增加有较好的相关性,不成熟粒细胞的数量可间接反映干／祖细胞的数量,故MNC与不成熟粒细胞数结合可作为allo-PBSCT的剂量阈标准。     

外周血造血干细胞的动员、采集和移植后造血重建

目的对13例健康供者及患者外周血造血干细胞的动员和采集效果进行分析。方法健康供者用rhG-CSF动员;患者采用强化疗加rhG-CSF或和rhGM-CSF动员,COBESpectro血细胞分离机全自动造血干细胞程序采集单个核细胞。结果13例移植中8例一次采集成功,1例(即首例)采集4次,余者采集2次。采集后的MNC6×108/kg～12×108/kg,CD3+4细胞3×106/kg～34×106/kg。移植后全部患者造血均获重建,异基因外周血造血干细胞移植一个月后DNA指纹图均提示植活。结论rhG-CSF可作为健康供者的安全有效动员剂,强化疗加rhG-CSF或/和rhGM-CSF是白血病、淋巴瘤自体外周血造血干细胞移植的有效动员方法之一。     

化疗联合单一剂量rhG-CSF用于自体外周血造血干细胞移植的临床研究

背景与目的:总结广东省干细胞多中心研究协作组自1999年6月至2001年12月间55例自体外周血造血干细胞移植治疗造血系统恶性疾病的资料,对化疗联合单一剂量rhG-CSF用于自体外周血造血干细胞移植前动员及移植后造血重建的效果进行研究和评价。方法:全部病例(急性髓细胞性白血病28例,急性淋巴细胞性白血病9例,非霍奇金淋巴瘤14例,其他4例)采用化疗+重组粒系集落刺激因子(rhG-CSF,格拉诺赛特)联合动员方案,其中白血病患者主要采用EA方案,恶性淋巴瘤患者主要采用以CTX为主的方案。rhG-CSF用量为250μg/d,WBC升至>4×109/L后,连续1～2天采集PBSC。移植后+3天开始使用rhG-CSF250μg/d,并观察造血重建情况。结果:动员所需的时间即自化疗开始至采集的平均时间为(18.08±3.63)天,rhG-CSF平均应用剂量为4.15μg·(kg·d)-1,应用时间平均7.12天。55例患者平均采集1.38次,采集到的MNC细胞数为(4.09±1.69)×109/kg,CD34+细胞平均值为8.5×106/kg,CFU-GM平均为(6.1±5.8)×105/kg。WBC恢复至>1.0×109/L及中性粒细胞绝对值>0.5×109/L的中位天数分别为10天和10.5天,全部移植患者均获满意的造血重建。结论:我们采用的EA和以CTX为主的化疗联合单一剂量rhG-CSF,是一种安全有效的动员自体外周血造血干细胞的方法,单一剂量rh     

正常供者外周血造血干细胞动员效果及影响因素

 目的　探讨粒细胞集落刺激因子（G-CSF）动员正常供者外周血造血干细胞的效果、毒副作用及影响因素。方法　对59名异基因造血干细胞正常供者采用G-CSF 皮下注射3 ~ 5 d,使用COBE Spectra血细胞分离机采集外周血干细胞,流式细胞术检测采集物中CD+34细胞数。结果　所有供者第一次采集的单个核细胞（MNC）及CD+34细胞量平均值分别为4.4（1.12～13.06）×108／kg供者体重及3.78（1.14～12.92）×106／kg供者体重。患者不良反应轻微。男性供者、年龄小于45岁者及采集前白细胞计数高者采集所得CD+34细胞数较高。结论　G-CSF作为正常供者动员剂安全有效。患者性别、年龄及采集前白细胞计数可作为预测因素。     

环磷酰胺或足叶乙甙联合粒细胞集落刺激因子动员自体外周血造血干细胞的对照研究

背景与目的:通过动员采集获得高质量的自体外周血造血干细胞(autologousperipheralbloodstemcell,APBSC)是造血干细胞移植成功的关键,环磷酰胺(cyclophosphamide,CTX)联合重组人粒细胞集落刺激因子(recombinedhumangranulocytecolony-stimulatingfactor,rhG-CSF)是APBSC经典的动员方案,足叶乙甙(etoposide,VP-16)联合rhG-CSF是近年来应用的另一个动员方案。本研究的目的是比较上述两种动员方案对恶性淋巴瘤和生殖细胞肿瘤患者APBSC的动员效果。方法:共有52例恶性实体瘤患者,其中CTX方案组26例,剂量为CTX3.5g/m2加rhG-CSF5μg·kg-1·d-1;VP-16方案组26例,VP-16的剂量随机采用1000mg/m2或1500mg/m2加rhG-CSF5μg·kg-1·d-1。两组均在白细胞(whitebloodcell,WBC)降至最低点时开始皮下注射rhG-CSF,直至采集结束前一天。当CTX组WBC恢复到2.5×109/L、VP-16组WBC恢复到5.0×109/L以上时开始连日采集APBSC,当累计采集的单个核细胞(mononuclearcell,MNC)≥5×108/kg或CD34+细胞≥2×106/kg时停止采集。患者经预处理后回输采集到的APBSC。比较两组动员采集过程中的血液学指标变化、采集细胞数量、造血重建时间、不良反应等。结果:CTX组患者化疗后外周血中WBC和血小板(platelet,PLT)降至最低值的时间明显早于VP-     

中剂量环磷酰胺为主的联合化疗加同一剂量G-CSF动员恶性血液病患者自体外周血干细胞

目的:观察中剂量环磷酰胺(CTX)为主的联合化疗加G蛳CSF对恶性血液病患者自体外周血造血干细胞(APBSC)的动员效果。方法:31例患者接受中剂量CTX 2.2 g/m2(1.8 g/m2~3.0 g/m2)联合VP16(600 mg ~ 800 mg)或Ara蛳C(1.0 g/m2 ~ 2.0 g/m2)化疗,WBC降至最低值后开始皮下注射G蛳CSF 300 μg/d直至采集结束。WBC≥(3.0~5.0)×109/L时开始采集,当单个核细胞(MNC)累计≥3.8×108/kg或CD+34细胞≥2.0×106/kg时停止采集。结果:采集次数为(2.9±1.0)次,G蛳CSF持续应用时间为(7.4±2.0)d,采集到的MNC细胞数为(5.53±2.54)×108/kg,CD+34细胞数为(9.46±7.24)×106/kg,CFU蛳GM(46.02±70.58)×104/kg。全部移植患者造血功能均获满意重建。结论:中剂量CTX为主的联合化疗加同一剂量G蛳CSF对血液病患者的APBSC动员是安全、有效的。     

外周血造血祖细胞检测对外周血干细胞动员采集的预测意义

 目的　通过血细胞分析仪检测外周血造血祖细胞（HPC）含量,探寻一种快速预测采集物干细胞含量、判断外周血造血干细胞采集时机的手段,并评价其应用意义。方法　对27例化疗联合粒细胞集落刺激因子（G-CSF）进行外周血造血干细胞动员采集者及17例单用G-CSF动员的采集者,利用血细胞分析仪检测外周血HPC计数,血液治疗和移植工程国际组织（ISHAGE）法检测外周血及采集物CD+34 细胞计数,并进行相关回归分析。对外周血HPC计数预测采集物CD+34 含量进行受试者工作曲线（ROC）分析。结果　无论化疗组或非化疗组,外周血HPC计数与CD+34 计数均具有线性相关性（r＝0.711,P＝0.000及r＝0.656,P＝0.004）。化疗组外周血CD+34 计数＝-0.829+0.648×外周血HPC计数;非化疗组外周血CD+34 细胞计数＝45.033+0.460×外周血HPC计数。化疗组患者外周血HPC计数与采集物CD+34 细胞量呈线性相关（r＝0.602,P＝0.001）,采集物CD+34 计数＝1.106+0.046×外周血HPC计数。以HPC≥85／μl预测采集物CD+34 计数≥5×106／kg的灵敏度为78 %,特异度为82 %。结论　在自体干细胞移植动员患者中,外周血HPC计数可部分替代外周血CD+34 计数预测采集效果,HPC≥85／μl为较好的预测阈值。     

大剂量多西他塞联合粒细胞集落刺激因子用于恶性实体瘤外周血造血干/祖细胞动员

目的观察大剂量多西他塞联合粒细胞集落刺激因子(G-CSF)在恶性实体瘤患者动员采集自体外周血造血干/祖细胞的有效性和安全性。方法30例恶性实体瘤患者入组,第一天采用多西他塞120mg/m2经静脉持续滴注3小时,在白细胞1.0×109/L左右时每天给予G-CSF5μg/kg,分早晚两次皮下注射,直至采集结束。结果在给予多西他塞后平均第(6.47±1.01)天白细胞降至1.0×109/L,皮下注射G-CSF平均(3.50±1.01)天即多西他塞动员后平均第(9.97±1.03)天开始外周血造血干/组细胞单采,每人每天采集一次,采集两天,共获CD34+细胞中位数3.49×106/kg(1.71×106～16.69×106/kg),其中CD34+细胞>2.0×106/kg的患者25例。不同采集时间获CD34+细胞数差异无统计学意义(P=0.651)。6例发生关节轻度疼痛,3例轻度腹泻,1例口腔粘膜炎。结论多西他塞120mg/m2联合G-CSF5μg/kg是恶性实体瘤患者动员采集自体干/祖细胞的有效安全方案。     

Efficacy and toxicity of lenograstim (glycosylated recombinant human granulocyte-colony stimulating factor) on peripheral blood stem cell harvest in healthy donors

Peripheral blood stem cell harvest with lenograstim (glycosylated rhG-CSF) was performed 12 times from 10 normal donors. Five micrograms/kg of lenograstim was administered subcutaneously twice a day (10 micrograms/kg/day) for 4 to 6 days, and apheresis was performed on day 5 to 7 depending on the collected CD 34+ cell counts. We collected a sufficient number of CD 34+ cells in 9 mobilizations from 7 donors less than 50 years of age, with a total number of collected CD 34+ cells in each mobilization of 22.1 +/- 6.5 x 10(7). In contrast, we could not obtain a sufficient number of CD 34+ cells in 2 mobilizations from 3 donors above 50 years of age, with a total number of collected CD 34+ cells of 9.8 +/- 3.3 x 10(7). Although all donors had adverse events in response to lenograstim administration, all of them were grade 2 or less toxicity. These results indicate that peripheral blood stem cell mobilization and apheresis by lenograstim is safe and well tolerated, but the risk of poor mobilization may become higher in donors more than 50 years of age.     

Isolation and transplantation of highly purified autologous peripheral CD34<Superscript>+</Superscript> progenitor cells: purging efficacy,hematopoietic reconstitution following high dose chemotherapy in patients with breast cancer: results of a feasibility study in Japan

BACKGROUND: High-dose chemotherapy with autologous stem cell support may have some therapeutic impact on certain groups of the patients with advanced breast cancer(BRCA). Since stem cell contamination by tumor cells might contribute to relapse, development of a tumor cell purging technique would improve the clinical outcome. The present study was undertaken to evaluate the purging efficacy of autologous mobilized CD34+peripheral stem cells in patients with breast cancer (BRCA) in an advanced stage or relapse. METHODS: CD34+cells were selected from autologous peripheral blood stem cells (PBSC) using a clinical scale of magnetic-activated cell sorting system (CliniMACS), followed by high-dose chemotherapy with transplantation of CD34+ selected cells. Amplification of cytokeratin 19 (CK19) and 20 (CK20) gene in leukapheresis products were measured to evaluate the performance of tumor cell elimination. RESULTS: Seven patients were entered into this study. After leukopheresis, 1 patient was dropped form this study due to poor mobilization. Among 6 patient, a total of 8 CD34+ selection was performed. The median purity and recovery rate of the CD34+ cells post selection was 85.1% (range 62.5-98.1%) and 74.2% (range 50.2-90.2%), respectively. After isolation of CD34+cells, the elimination rate in the logarithmic transformation of CK19 was 2.77 log, and that of CK20 were 2.43 log and 2.53 log. In 4 patients, high-dose chemotherapy was performed, followed by the transplantation of the isolated CD34+cells. Rapid neutrophil recovery, as well as platelet recovery was seen with a median time to reach 0.5 x 109/l neutrophils of 9 days(range 8-9), and 20 x 109/l platelets of 12 days (range 10-13). There was no treatment related death and no serious adverse events directly associated with the selection procedure or infusion of selected cells. CONCLUSIONS: The present study demonstrated that the CliniMACS system is a highly effective positive selection method and that a high purging efficacy could be obtained without compromising the hematopoietic reconstitution capacity of the graft in BRCA patients undergoing high-dose chemotherapy.     

High-dose chemotherapy followed by reinfusion of selected CD34+ peripheral blood cells in patients with poor-prognosis breast cancer: a randomized multicentre study.

Seventy-one patients with poor-prognosis breast cancer were enrolled after informed consent in a multicentre randomized study to evaluate the use of selected peripheral blood CD34+ cells to support haematopoietic recovery following high-dose chemotherapy. Patients who responded to conventional chemotherapy were mobilized with chemotherapy (mainly high-dose cyclophosphamide) and/or recombinant human granulocyte colony-stimulating factor (rhG-CSF). Patients who reached the threshold of 20 CD34+ cells per microl of peripheral blood underwent apheresis and were randomized at that time to receive either unmanipulated mobilized blood cells or selected CD34+ cells. For patients in the study arm, CD34+ cells were selected from aphereses using the Isolex300 device. Fifteen patients failed to mobilize peripheral blood progenitors and nine other patients were excluded for various reasons. Forty-seven eligible patients were randomized into two comparable groups. CD34+ cells were selected from aphereses in the study group. Haematopoietic recovery occurred at similar times in both groups. No side-effect related to the infusion of selected cells was observed. The frequency of epithelial tumour cells in aphereses was low (8 out of 42 evaluated patients), as determined by immunocytochemistry. We conclude that selected CD34+ cells safely support haematopoietic recovery following high-dose chemotherapy in patients with poor-prognosis breast cancer.     

联合化疗加重组人粒细胞集落刺激因子动员自体外周血干细胞

目的观察环磷酰胺(CTX)为主的联合化疗加重组人粒细胞集落刺激因子(rhG-CSF)对自体外周血干细胞(APBSC)的动员效果。方法CTX（2.5±1.0）g/m     

Efficacy of first-line bleomycin, etoposide, and cisplatin chemotherapy for peripheral blood stem cell mobilization in patients with germ cell cancer

Background We assessed the efficacy of first-line bleomycin, etoposide, and cisplatin (BEP) chemotherapy for the mobilization of peripheral blood stem cells (PBSC) in patients with testicular cancer, and analyzed the predictive factors indicating the optimal time of PBSC harvest. Patients and Methods A total of 29 aphereses, performed during first-line BEP chemotherapy between 1994 and 1996 for 10 patients with metastatic germ cell cancer were analyzed. The predictive value for the optimal time of PBSC harvest was determined by analysis of the correlation between the rate of each cellular component in peripheral blood, and the number of CD34-positive cells harvested. Results The median number of CD34-positive cells obtained at a single apheresis was 11.2×10⁶/kg (range, 0.14 to 47.9×10⁶/kg), and the median cumulative number of CD34-positive cells collected during first-line BEP chemotherapy per patient was 31.9×10⁶/kg (range, 9.7 to 75.5×10⁶/kg). The percentage of immature leukocytes (myelocytes plus metamyelocytes) was significantly correlated with the number of harvested CD34-positive cells. Conclusion Adequate amounts of CD34-positive cells can be harvested during first-line BEP chemotherapy for patients with germ cell cancer. The monitoring of the percentage of immature leukocytes might be useful in ascertaining the optimal time of apheresis.