Application of peripheral blood stem cells (PBSC) mobilized by recombinant human granulocyte colony stimulating factor for allogeneic PBSC transplantation and the comparison of allogeneic PBSC transplantation and bone marrow transplantation.

Recombinant human granulocyte colony stimulating factor (rhG-CSF)-mobilized peripheral blood stem cells (PBSC) are now widely used for allogeneic PBSC transplantation (alloPBSCT). Large numbers of hematopoietic progenitor cells mobilized by rhG-CSF would be considered equivalent or better than bone marrow (BM) cells and would be used as an alternative to BM for allogeneic hematopoietic stem cell transplantation. The complications associated with the administration of rhG-CSF and apheresis in PBSC collection in formal donors are well tolerated and usually acceptable in the short term but some hazardous adverse events such as splenic rupture and cardiac arrest are reported although the incidence is very low. Protective means and stopping rules for safe donation in the collection of PBSC are established. The characteristics of PBSC were clarified; the expression of some adhesion molecules such as CD49d on CD34 positive cells of PBSC have been shown to be low compared to BM stem cells. In alloPBSCT compared with allogeneic BM transplantation (alloBMT), the incidence and frequency of graft versus host disease (GVHD) is of concern because high number of T lymphocytes are infused in alloPBSCT. The incidence and severity of acute GVHD are not increased but chronic GVHD is higher in alloPBSCT compared with alloBMT. The outcome of alloPBSCT and BMT are almost equivalent and conclusive results regarding survival are not yet available.     

CD41+ and CD42+ hematopoietic progenitor cells may predict platelet engraftment after allogeneic peripheral blood stem cell transplantation

The objective of this study was to quantify subpopulations of CD34+ cells such as CD41+ and CD42+ cells that might represent megakaryocyte (MK) precursors in peripheral blood stem cell (PBSC) collections of normal, recombinant human granulocyte‐colony stimulating factor (rhG‐CSF) primed donors and to determine whether there is a statistical association between the dose infused megakaryocytic precursors and the time course of the platelet recovery following an allogeneic PBSC transplantation. Twenty‐six patients with various hematologic malignancies transplanted from their HLA identical siblings between July 1997 and December 1999 were used. All patients except one with severe aplastic anemia who had cyclophosphamide (CY) alone received busulfan‐CY as preparative regimen and cyclosporine‐methotrexate for GVHD prophylaxis. Normal healthy donors were given rhG‐CSF 10 μg/kg/day subcutaneously twice daily and PBSCs were collected on days 5 and 6. The median number of infused CD34+, CD41+ and CD42+ cells were 6.61 × 10⁶/kg (range 1.47–21.41), 54.85 × 10⁴/kg (5.38–204.19), and 49.86 × 10⁴/kg (6.82–430.10), respectively. Median days of ANC 0.5 × 10⁹/L and platelet 20 × 10⁹/L were 11.5 (range 9–15) and 13 (8–33), respectively. In this study, the number of CD41+ and CD42+ cells infused much better correlated than the number of CD34+ cells infused with the time to platelet recovery of 20 × 10⁹/L in 26 patients receiving an allogeneic match sibling PBSC transplantation (r = ?0.727 and P < 0.001 for CD41+ cells, r = ?0.806 and P < 0.001 for CD42+ cells, r = ?0.336 and P > 0.05 for CD34+ cells). There was an inverse correlation between the number of infused CD41+ and CD42+ cells and duration of platelet engraftment. Therefore, as the number of CD41+ and CD42+ cells increased, duration of platelet engraftment (time to reach platelet count of ≥ 20 × 10⁹/L) shortened significantly. Based on this data we may conclude that flow cytometric measurement of CD41+ and CD42+ progenitor cells may provide an accurate indication of platelet reconstitutive capacity of the allogeneic PBSC transplant. J. Clin. Apheresis. 16:67–73, 2001. © 2001 Wiley‐Liss, Inc.     

同胞异基因外周血干细胞移植治疗白血病研究

本研究观察异基因外周血干细胞移植 (PBSCT)治疗白血病的疗效与副作用。所选供者均为HLA A、B、DR位点与患者完全相同的同胞兄弟姐妹。供者经 2 5 0 μg/kgrhG CSF动员 5天后 ,单采外周血干细胞 1- 2次。 4例白血病病人经过改良Bu/Cy预处理方案 ,输入单个核细胞 6.78× 10 8/kg± 1.96× 10 8/kg( 5× 10 8/kg - 8.67×10 8/kg) ,其中CD3 4 为 15 .0 2× 10 6 /kg± 8.93× 10 6 /kg ( 5 .3× 10 6 /kg - 2 4 .2 3× 10 6 /kg) ,用MTX CsA MMF预防急性移植物抗宿主病 (aGVHD)。结果表明 :移植前 2天到移植后 2天白细胞降至最低 ,移植后 11- 17天中性粒细胞 >0 .5× 10 9/L ,移植后 11到 5 5天血小板 >5 0× 10 9/L。 4例中 2例出现aGVHD ,2例出现cGVHD ,2例出现感染。移植后 2 8天复查骨髓显示造血恢复 ,STR位点的DNA检查显示供者细胞生长。结论 :采用改良Bu/Cy方案和用MTX CsA MMF预防aGVHD方案进行PBSCT治疗白血病是一种安全、可靠的治疗方法。     

The impact of the CD34+ cell dose on engraftment in allogeneic peripheral blood stem cell transplantation

Forty-five patients who underwent allogeneic peripheral blood stem cell transplantation (PBSCT) were evaluated in order to investigate any relationship between CD34+ cell dose given and hematological recovery. Granulocyte counts > 1.0 x 10(9)/L and platelet > 50 x 10(9)/L were considered as hematological recovery. Three different regimens were used for mobilization, by adjusting the recombinant granulocyte colony stimulating factor (rhG-CSF, Roche) dose. The first group (n = 3), whose donors mobilized with 5 micrograms/kg/d s.c. rhG-CSF received a mean of 5.9 x 10(6)/kg (95% confidence interval for mean (CI); 2.4-9.3) CD34+ cells. The second group (n = 37), mobilized with 10 micrograms/kg/d s.c. rhG-CSF and the third group (n = 5) mobilized with 15 micrograms/kg/d s.c. rhG-CSF, received a mean of 5.7 x 10(6)/kg (95% CI; 4.6-6.75) and 6.56 x 10(6)/kg (95% CI; 4.57-8.55) CD34+ cells, respectively. CD34+ cell dose was 5.82 x 10(6)/kg (95% CI; 4.97-6.68) for all the patients. All patients received rhG-CSF from day +1 until attaining granulocyte count > 1.0 x 10(9)/L for three consecutive days. Median granulocyte and platelet engraftment days for the whole group was 15 (range; 11-44) and 14 (11-54) days respectively. There was a close correlation (r = -0.301, p < 0.05) between the CD34+ cell dose and granulocyte recovery for the whole group. When these analyses were performed separately within groups, this correlation was also found significant for the first group (r = -0.99, p < 0.05) for granulocyte recovery. On the contrary the same analysis did not reach significance for the other groups, nor for platelet recovery for the whole group (r = 0.039, p = 0.821). We calculated a minimum dose of 4 x 10(6)/kg CD34+ cells for a safe alloPBSCT. There was no difference between patients who received more than 5 x 10(6)/kg CD34+ cells, and those who received more than 2 x 10(6)/kg and less than 5 x 10(6)/kg CD34+ cells. In conclusion, we have demonstrated a correlation between the CD34+ cell dose given and faster hematological recovery for alloPBSCT patients.     

Mobilization of peripheral blood stem cells with paclitaxel and rhG-CSF in high-risk breast cancer patients

Preclinical studies have demonstrated the rapid and efficient mobilization of hematopoietic peripheral blood stem cells (PBSC) in a mouse model using the combination of paclitaxel with recombinant human granulocyte colony-stimulating factor (rhG-CSF). On the basis of these results, a clinical trial was initiated using rhG-CSF with paclitaxel for PBSC mobilization in high-risk breast cancer patients. The mobilized PBSC were evaluated for CD34(+) cell number, mononuclear cell content, and clonogenic potential. One-hundred and seventeen breast cancer patients received paclitaxel (300 mg/m(2)) administered as a 24-h continuous intravenous infusion. Forty-eight hours after completing paclitaxel, rhG-CSF (5 microg/kg) was initiated and continued until completion of PBSC collection. Leukapheresis was initiated once the white blood cell count reached 1.0 x 10(9)/L. Each collection was evaluated for the numbers of mononuclear cells (MNC) and CD34(+) cells. Clonogenic potential was enumerated using colony-forming units-granulocyte-macrophage (CFU-GM) and burst-forming units-erythroid (BFU-E). Patients receiving paclitaxel with rhG-CSF mobilized a large number of mononuclear cells/apheresis (mean, 3.7 x 10(8); range, 3.3-4.1) and CD34(+) cells/apheresis (mean, 7.2 x 10(6); range, 6.1-8.4). The average number of leukophereses needed was 1.8 (mean, range 1.6-2.0). Colony growth was normal with 178.9 x 10(5) and 214.8 x 10(5) colonies counted in CFU-GM and BFU-E assays, respectively. Patients engrafted platelets and neutrophils on day 10 following transplantation. In conclusion, PBSC mobilization with paclitaxel and rhG-CSF results in a large number of mononuclear cells and CD34(+) cells with normal clonogenic potential. The cells engraft normally following high-dose chemotherapy and autologous stem cell transplantation in high-risk breast cancer patients. These results demonstrate that paclitaxel with rhG-CSF is an efficient mobilizing agent in high-risk breast cancer patients.     

Factors influencing mobilization and engraftment in patients with metastatic breast cancer undergoing PBSC transplantation.

Factors influencing mobilization and engraftment of PBSC were analyzed in 38 patients with metastatic breast cancer who were undergoing PBSC transplantation. None of these patients had had previous chemotherapy for metastatic disease. PBSC were mobilized with cyclophosphamide (CY) and G-CSF (n = 21) or CY and etoposide (CY-etoposide) and G-CSF (n = 17). All received cyclophosphamide 6000 mg/m2, thiotepa 500 mg/m2, and carboplatin 800 mg/m2 (CTCb) as preparative regimen. PBSC infusion was followed by G-CSF at 5 microg/kg in 30 patients or 10 microg/kg in 8 patients. A median number of 27 x 10(6) CD34+ cells/kg was obtained with a median of four aphereses. Previous chemotherapy, radiation therapy, marrow disease, time from previous chemotherapy to mobilization, and type of mobilization regimen did not have a statistically significant effect on collection efficiency (CE). CE was defined as the total number of CD34+ collected/number of collections. Engraftment was rapid, with patients reaching a neutrophil count of 0.5 x 10(9)/L a median of 9 days (range 7-23) and a platelet count of 20 x 10(9)/L a median of 12 days (range 8-28) after transplantation. Shorter times to platelet recovery were associated with a higher number of CD34+ cells infused (p = 0.012), CY mobilization (p = 0.033), and a lower number of prior chemotherapy cycles (p = 0.022). When the number of CD34+ cells was included in the proportional hazard model, no other variables were found to be significant predictors of platelet engraftment. Time to neutrophil recovery was negatively associated with the dose of G-CSF used after transplantation (p = 0.036) CD34 cell dose is an important predictor of engraftment kinetics. A posttransplant dose of G-CSF improves neutrophil recovery. For patients with metastatic breast cancer and no previous chemotherapy for metastatic disease, we have no evidence for a difference between CY and CY-Etoposide as the mobilization regimen.     

Peripheral blood stem cell mobilization for high-dose chemotherapy.

Several studies have clearly documented a more rapid hematopoietic recovery with growth factor-mobilized PBSC than with bone marrow. Time to engraftment for neutrophils and platelets average 8-12 days in contrast to 2-4 weeks after bone marrow. This rapid hematopoietic recovery with PBSC has decreased the duration of hospitalization, transfusion requirements, and costs. Although growth factors alone may mobilize enough PBSC for high-dose chemotherapy, administration of growth factor after submyeloablative chemotherapy increases the yield of CD34+ cells. Based on the current data, CD34+ cell content of PBSC appears to be the single most powerful predictor of hematopoietic recovery. Infusion of > or =5 x 10(6) CD34+ cells/kg is associated with a rapid engraftment of neutrophils and platelets, although successful engraftment has also been reported with infusion of 2.5-5 x 10(6) CD34+ cells/kg. Age, prior radiotherapy, marrow involvement, and prior chemotherapy regimens are important factors influencing the yield of stem cells. Therefore, using these pa-rameters, we may identify the patients who will fail to mobilize sufficient numbers of PBSC before collection and use new strategies for stem cell mobilization. Because of the ease of collection and rapid engraftment after myeloablative therapy, PBSC have replaced bone marrow for autologous transplantation and may supplant bone marrow for allogeneic transplantation in the near future.     

FBC预处理方案在HLA半相合异基因外周血干细胞移植中的应用

目的　观察降低预处理强度对HLA半相合异基因干细胞植入的影响。方法　用氟达拉宾 (30mg m2 × 6d)、白消安 (4mg kg× 2d)、环磷酰胺 [(30～ 6 0 )mg kg× 2d]组成FBC方案。将 12例白血病患者分成两组 ,HLA完全相合移植组 4例 ,移植前患者处完全缓解状态 ;HLA半相合移植组 8例 ,皆为难治性白血病 ,其中 1例HLA 3个位点不合 ,6例二个位点不合 ,1例一个位点不合 ,接受本处理方案后 ,进行G CSF动员的异基因外周血干细胞移植 ,并于移植后第 30天 (+30天 )、+6 0天、+90天输注供者淋巴细胞 ,观察异基因干细胞植入和长期植入情况。结果　HLA半相合组患者平均接受 4.87× 10 8 kg供者外周血单个核细胞 ,其中CD3 4+细胞平均数为 4.5 8× 10 6 kg,HLA全相合组平均接受 4.85× 10 8 kg供者外周血单个核细胞 ,其中CD3 4+细胞 4.47× 10 6 kg。HLA半相合组 7例白细胞升至 1.0×10 9 L的平均时间为 2 9d(11～ 90d) ,血小板升至 2 0× 10 9 L的时间为 36d(14～ 96d) ,1例HLA 3个位点不合的患者移植失败 ,但该患者于 +5 0天恢复自体造血。而HLA全相合的 4例患者白细胞升至 1.0×10 9 L平均需 14d(11～ 18d) ,血小板升至 2 0× 10 9 L平均需 15d(11～ 18d)。经STR PCR检测 ,两组患者除 1例表现为混合嵌合体 ,其余均呈完     

Peripheral blood stem cell apheresis for allogeneic transplants: Ibni Sina experience

BACKGROUND: The rate of utilizing peripheral blood stem cells (PBSC) as a source for allogeneic stem cells is growing rapidly. We aimed to demonstrate our 4 years experience as the largest apheresis center in Turkey and analyzed the content of the apheresis material. PATIENTS AND METHODS: From 1998 to the end of April 2002, 151 leukopheresis procedures were performed on 116 healthy donors (M/F:66/50) with a median age of 30 years (14-53). The HLA identical sibling donors received rhG-CSF 10 microg/kg/day sc. for 4 days and at the 5th day leukopheresis was started until collecting >4 x 10e6/kg CD34+ cells. Two times the donors' total blood volume was processed in 195 min (178-245) on continuous flow cell separators using peripheral venous access. RESULTS: Preapheresis WBC was 51.5 x 10e9/L (range, 13.11-91.3). Mono nuclear cell, CD34 and CD3 quantity of the harvest material were 5.35 x 10e8/kg (range, 0.45-23.46), 6.4 x 10e6/kg (range, 2.49-33.27) and 2.79 x 10e8/kg (range, 0.46-30.95), respectively. We were able to reach the target CD34 count after 1st cycle in 39% and 2nd cycle 61% of the procedures. In all donors with a peripheral blood CD34 count >80/mcl we succeeded to collect enough stem cells with only one leukopheresis. CONCLUSION: Collection of peripheral blood stem cells with continuous flow cell separators is well tolerated, with no mobilization failures or poor mobilizers. We collected high values of CD34+ cells (med. 6.4 x 10e6/kg) at the expense of high CD3+lymphocytes (med. 2.79 x 10e8/kg), which may increase the risk of acute and chronic GVHD after allogeneic hemapoietic cell transplantation.     

G-CSF预处理供者单倍体相合骨髓移植加用CD25 单克隆抗体预防急性GVHD的临床研究

探索单倍体相合未去T细胞骨髓移植应用CD25单克隆抗体预防急性移植物抗宿主病(GVHD)的疗效。在1999年2月至2001年11月期间，对28例白血病病人进行HLA2—3个位点不合骨髓移植，其中2000年11月前完成的不使用CD25单杭的15例为对照组，此后完成的13例为本方案组，两组供均接受粒细胞集落刺激因子(G—CSF，Lenograstim)250μg／d，皮下注射，连用7天后采髓；环抱菌素A，氨甲蝶呤，抗胸腺细胞球蛋白和霉酚酸酯(MMF)联合用于GvHD预防。研究组病例在移植前2小时和移植后第4天加用CD25单克隆抗体(Basiliximab)进一步预防GvHD；并对两组移植后植入，GvHD发生和无病存活情况进行比较。结果表明，28例患移植后均植入成功，植入直接证据的检测证实完全供造血，两组造血重建时间比较无显性差异(P＞0.05)；未用抗体的对照纽急性Ⅱ—IV度GvHD发生率33.3％，本方案组加用CD25单克隆抗体，无1例发生急性Ⅱ—IV度GVHD，两组之间差异有显性(P＜0．05)；慢性GvHD发生均为局限性，两组的差异无显性(P＞0.05)。对照组中位随访26(15—36)月，活存9／15例，本方案组中位随访8(3—15)月，活存12／13例，Kaplan—Meier存活曲线分析与对数秩检验，本方案组1年无病生存率明显高于对照组(P＜0．05)。结论：单倍体相合骨髓移植加用CD25单克隆抗体有利于急性重度GvHD的预防，显提高单倍体相合骨髓移植无病生存率。     

自体纯化CD34+细胞移植治疗严重自身免疫性疾病的初步研究

目的探讨自体外周血CD34+细胞移植治疗严重自身免疫性疾病的干细胞动员、细胞采集和分选、预处理和并发症处理等问题.方法 10例重度自身免疫性疾病患者接受自体外周血CD34+细胞移植治疗.采用环磷酰胺(CTX)+rhG-CSF方案动员外周血干细胞,并以CliniMACS细胞分选仪分选CD34+细胞,适时用CTX+抗胸腺细胞球蛋白(7例)或CTX+全身照射(3例)两种预处理方案后,进行CD34+细胞回输的方法治疗.结果经CTX+rhG-CSF方案动员并以CliniMACS细胞分选仪分选后,可获得(1.98±0.95)×108的CD34+细胞,其纯度为(91.4±10.6)%,回收率为(60.5±19.8)%.在回输(2.14±1.05)×106/kg的CD34+细胞后,ANC ≥0.5×109/L的时间为(8.6±2.5)d,血小板升至20×109/L的时间为(9.0±5.2)d.在造血恢复后,所有CD3+细胞、CD19+细胞和CD16+CD56+细胞均未恢复至移植前状态.在造血和免疫抑制时,巨细胞病毒感染的发生率较高.2例患者死于移植相关并发症.所有患者近期疗效满意,6例系统性红斑狼疮患者DAI评分由移植前的平均17分降为移植后的4分;类风湿关节炎患者DAS28评分由6.4分降至1.8分;干燥综合征患者的症状和体征均明显缓解.结论对常规治疗无效的严重自身免疫性疾病,自体外周血CD34+细胞移植是可选择的治疗方法之一.     

低剂量rhG-CSF对56例非血缘供者外周造血干细胞动员

本研究观察低剂量人重组粒细胞集落刺激因子(rhG-CSF)对非血缘健康供者的影响,探讨用于中华造血干细胞捐赠者资料库提供的非血缘健康供者外周造血干细胞动员方案。56例非血缘健康供者接受rhG-CSF 5μg/(kg.d)皮下注射,在动员第4、5两天或第5、6两天采集干细胞,观察动员效果及不良反应,检测动员前后血常规指标、CD3+、CD4+、CD8+和CD20+细胞比例;对采集物进行单个核细胞(MNC)和CD34+细胞计数;对所有供者随访至2006年5月31日。结果显示:在rhG-CSF动员过程中出现1级毒副作用(按WHO分级标准):腰背酸痛17.9%(10/56)、焦虑失眠8.9%(5/56)、疲乏4.5%(3/56)等,无需特殊处理,无需终止动员。第4、5两天采集和第5、6两天采集所得的MNC分别是(5.95±1.52)×108/kg和(7.19±2.12)×108/kg;CD34+细胞分别是(3.03±1.09)×106/kg和(7.92±2.50)×106/kg。血红蛋白水平、血小板量、CD3、CD4、CD8、CD20百分比动员前后无变化。结论:5μg/(kg.d)rhG-CSF用于非血缘健康供者的动员是安全而有效的。     

异基因外周血造血干细胞移植治疗骨髓增生异常综合征23例疗效分析

目的观察异基因外周血造血干细胞移植(allo-PBSCT)治疗骨髓增生异常综合征(MDS)的疗效。方法对23例 MDS 患者行 HLA 相合的同胞间 allo-PBSCT,供者外周血采集前给予G-CSF 动员。输注外周血单个核细胞8.25(4.50～22.36)×10~8/kg,CD34~ 细胞5.59(1.57～12.22)×lO~6/kg 移植物抗宿主病(GVHD)的预防给予环孢菌素 A 加短疗程甲氨蝶呤,移植后1天( 1天)～ 28天给予霉酚酸酯。结果 23例患者中22例造血完全重建。中性粒细胞>1.0×10~9/L 和 BPC>50 × 10~9/L 的时间分别为 13( 11～ 17)天和 30( 13～ 102)天。移植相关死亡2例,移植后复发3例,现18例存活。 Kaplan-Meier 分析显示无病生存率、复发率分别为(77.8±8.7)%和(14.4±7.5)%。结论 Allo-PBSCT 治疗 MDS 治愈率高,复发率低。     

重组人白介素11联合粒细胞集落刺激因子动员的自体外周血造血干细胞移植

本研究旨在探索重组人白介素11（rhIL—11）联合重组人粒细胞集落刺激因子（rhG—CSF）动员外周血造血干细胞进行自体外周血干细胞移植的作用。16例预行自体外周血干细胞移植的非霍奇金淋巴瘤及急性髓系白血病患者随机分为实验组（rhIL-11联合rhG—CSF动员）及对照组（rhG—CSF动员），两组均在动员性化疗后血象下降至最低值有回升迹象时应用rhIL—11及rhG—CSF；rhG—CSF5μg/（kg·d）动员中位时间5．5天，rhIL—1150μg/（kg·d）动员中位时间4天；动员后观察外周血白细胞和血小板计数，以及干细胞采集物单个核细胞、CD34^＋细胞、CFU—GM集落数的变化；按常规进行自体外周血干细胞移植后，观察粒细胞及血小板植活时间及单采血小板输注量。结果显示：实验组及对照组动员后外周血白细胞和血小板计数，以及干细胞采集物单个核细胞、CD54^＋细胞及CFU—GM集落数无显著性差异（P〉0．05）。自体外周血干细胞移植后，实验组中性粒细胞数≥0.5×10^9／L的中位时间为10．5天，对照组中为13天．实验组比对照组提前2.5天（P〈0．05）。实验组血小板数≥20×10^9／L的中位时间为11.5天，对照组为13天，实验组比对照组提前1.5天（p〈0．05）。实验组输注单采血小板中住数为3.5单位，对照组为5单位，实验组比对照组减少1．5单位（P〈0．05）。实验组使用动员剂的不良反应主要有低热、乏力、感冒样症状、食欲不振、头晕、肌肉酸痛等，对照组仅出现低热，患者对以上症状均可以耐受，停药后症状自行消失。结论：rhIL—11联合rhG—CSF动员外周血造血干细胞安全有效，在自体外周血干细胞移植后造血重建较快，单采血小板输注量少。     

Large-volume leukapheresis using femoral venous access for harvesting peripheral blood stem cells with the Fenwal CS 3000 Plus from normal healthy donors: predictors of CD34+ cell yield and collection efficiency

The current paper reports on the predicting factors associated with satisfactory peripheral blood stem cell collection and the efficacy of large-volume leukapheresis (LVL) using femoral vein catheterization to harvest PBSCs with Fenwal CS 3000 Plus from normal healthy donors for allogeneic transplantation. A total of 113 apheresis procedures in 57 patients were performed. The median number of MNCs, CD3+ cells, and CD34+ cells harvested per apheresis was 5.3 x 10(8)/kg (range, 0.3-11.0 x 10(8)/kg), 3.0 x 10(8)/kg (range, 0.2-6.6 x 10(8)/kg), and 7.9 x 10(6)/kg (range, 0.1-188.9 x 10(6)/kg), respectively. The median collection efficiency of MNCs and CD34+ cells was 49.8% and 49.7%, respectively. A highly significant correlation was found between the collected CD34+ cell counts and the pre-apheresis WBC counts in the donors (P = 0.013), and between the collected CD34+ cell counts and the pre-apheresis peripheral blood (PB) CD34+ cell counts (P<0.001). Harvesting at least >4 x 10(6)/kg CD34+ cells from the 1st LVL was achieved in 44 (77.2%) out of 57 donors and in 19 (90.5%) out of 21 donors with a PB-CD34+ cell count of >40/microl. There was no significant difference in the harvested MNC and CD34+ cell counts between the 1st and 2nd apheresis. The catheter-related complications included catheter obstruction (n = 2) and hematoma at the insertion site (n = 3). Accordingly, LVL using femoral venous access for allogeneic PBSC collection from normal healthy donors would appear to be safe and effective.     

异基因造血干细胞移植后供者外周血造血干细胞输注预防高危白血病复发

目的评估重组人粒细胞集落刺激因子(G-CSF)动员的供者外周血造血干细胞输注 (GPBSCI)作为一种早期过继性免疫治疗方法的有效性和安全性。方法 12例高危白血病患者同胞配型相合异基因造血干细胞移植(allo-HSCT)后接受了G-CSF动员的预防性GPBSCI。allo-HSCT前患者的诊断包括2例Ph 急性淋巴细胞白血病首次完全缓解(ALL-CR1),1例ALL-CR2,1例ALL合并顽固中枢神经系统白血病(CNSL),1例急性髓系白血病(AML)复发,1例AML合并CNSL,1例 AML-CR3,4例进展期慢性粒细胞白血病(CML)及1例骨髓增生异常综合征-难治性贫血伴幼稚细胞增多型(MDS-RAEB)。结果 12例患者共接受了16次GPBSCI,其中移植后90天( 90天)前接受 GPBSCI 5次,输注的单个核细胞(MNC)及CD3 细胞中位数分别为1．00(0．95-1．24)×108/kg和 0．53(0．39-0．63)×108/kg。 90天后接受GPBSCI 11次,输注两类细胞中位数分别为2．27(1．00- 4．30)×108/kg和1．15(0．55-2．10)×108/kg。输注后4例患者发生了Ⅰ或Ⅱ度急性移植物抗宿主病(GVHD),1例患者发生Ⅲ度急性GVHD。7例患者发生了慢性GVHD,其中4例为广泛型。2例患者未发生输注相关GVHD。未观察到GPBSCI相关的全血细胞减少。中位随访563(415-728)d,12 例高危白血病患者中有10例无病存活,2例死于白血病复发。结论预防性GPBSCI可以增强移植物抗白血病作用,相关不良反应小,可能成为改善高危白血病allo-HSCT预后的安全有效的手段。     

HLA相合同胞脐血移植治疗急性髓系白血病成功——国内首例报告

目的 研究脐血移植（ ＵＣＢＴ） 对恶性血液病的根治性治疗; 观察其长期造血重建和移植物抗宿主病（ＧＶＨＤ） 及移植并发症发生情况。方法 给１ 例急性粒细胞白血病完全缓解期的１１ 岁男性患儿移植ＨＬＡ 配型相合的同胞脐血。预处理选用ＢＵ／ＣＹ 方案（ 马利兰４ ｍ ｇ ·ｋｇ －１ ·ｄ － １ ×４ ,环磷酰胺６０ ｍｇ ·ｋｇ － １ ·ｄ － １ ×２） 。ＧＶＨＤ 预防用环孢霉素Ａ（ＣｓＡ） 。移植有核细胞为０ ．３５ ×１０８／ｋｇ ,ＣＦＵＧＭ １ ．８２ ×１０４／ｋｇ ,ＣＤ３４ ＋ 细胞２ ．０４ ×１０５／ｋｇ 。结果 ＋ １４ 天骨髓显示造血重建;＋ ２１ 天中性粒细胞（ＡＮＣ） ＞ １ ．０ ×１０９／Ｌ;＋ ６０ 天ＤＮＡ 位点检测显示移植物植入成功;＋ １００ 天受者血型由Ｏ 型转为供者血型Ｂ 型。随访３３０ 天,患者情况正常,未发生急、慢性ＧＶＨＤ。结论本例为国内首例异基因脐血移植治疗急性粒细胞白血病获得成功;开展同胞ＨＬＡ 相合脐血移植安全、有效,适合中国国情,值得深入研究。     

无关供者脐血造血干细胞移植治疗一例成人重型再生障碍性贫血

目的　研究无关供者脐血造血干细胞移植 (unrelatedallo CBSCT)治疗成人重型再生障碍性贫血 (再障 )。方法　用HLA配型 6个位点全相合无关供者allo CBSCT治疗 1例成人重型再障患者。脐血由广州市脐血库提供 ,输入的单个核细胞 (MNC)数为 1.89× 10 7/kg(按患者体重 6 0kg计 ) ,CD34 阳性细胞占 0 .0 0 9,CFU GM为 1.8× 10 4 /kg。预处理方案由环磷酰胺 (CTX)和抗淋巴细胞球蛋白 (ALG)组成 ,CTX总用量为 6 0mg/kg;ALG按 12 0mg/kg给予。用甲氨蝶呤 (MTX)和环孢菌素A(CsA)预防移植物抗宿主病 (GVHD) ,CsA用至 8个月逐渐减量停药。结果　输入脐血后白细胞最低值为 0 .6× 10 9/L ,血小板最低值为 12 .0× 10 9/L ,中性粒细胞于移植后第 10天大于 0 .5× 10 9/L ,血小板于移植后第 2 0天大于 5 0 0× 10 9/L。移植后 8个月时出现皮肤局限性GVHD。微卫星法DNA指纹图示稳定的供、受体混合嵌合状态 ,但迄今ABO血型仍为受者型。结论　此例为国内首例采用allo CBSCT治疗成人重型再障 ,并获得成功 ,脐血造血干细胞已长期植入。     

CD34+ cell enrichment for autologous peripheral blood stem cell transplantation by use of the CliniMACs device

Several devices for selection of CD34+ peripheral blood stem cells (PBSC) have been used during the last years for reducing tumor cell contamination of the graft. The new CliniMACS system (magnetic-activated cell separation system by Miltenyi Biotech GmbH, Bergisch-Gladbach, Germany) was recently approved for clinical use in Europe. To evaluate its purging efficiency and engraftment data in the autologous transplant, PBSC from 28 adult patients with various malignant diseases (non-Hodgkin's lymphoma, n = 17; chronic lymphocytic leukemia, n = 5; multiple myeloma, n = 4; acute lymphocytic leukemia, n = 1; medulloblastoma, n = 1) were mobilized by chemotherapy and granulocyte colony-stimulating factor (G-CSF) (10 microg/kg per day). Thirty leukapheresis products from 28 patients with a median of 4.4 x 10(8) nucleated cells/kg body weight (bw)(range 0.6-10.8 x 10(8)/kg bw) and a median of 7.1 x 10(6) CD34+ cells/kg bw (range 2.8 to 18.8 x 10(6)/kg bw) were selected using the Cobe spectra cell separator (Cobe BCT Inc., Lakewood, CO). After the CliniMACS procedure, the median yield of CD34+ selected cells was 4.5 x 10(6)/kg (range 2.2-11.1 X 10(6)/kg bw) with a median recovery of 69.5% (range 46.9-87.3%) and a median purity of 97.7% (range 89.4-99.8%). The procedure did not alter viability of selected cells, which was tested by propidium iodide staining. So far, purified PBSC were used for autologous transplantation in 15 out of 28 patients after total body irradiation and/or high-dose chemotherapy. Median time to reach an absolute neutrophil count > 500/microl was 12 days (range 10-18 days), platelet recovery >50,000/microl occurred at day + 16 (range 11-22). With a median follow-up time of 12 months (range 3-19), 5 patients died of relapse. We confirmed the feasibility and safety of the CliniMACS CD34+ cell enrichment procedure in adult patients with autologous PBSC transplantation.     

Effect of filgrastim administration for steady-state mobilization of peripheral blood stem cells.

To obtain a better (optimal) schedule of peripheral blood stem cell (PBSC) collection by steady-state granulocyte colony-stimulating factor administrations for autologous or allogeneic transplantations, we compared the effect of doses of filgrastim (8 microg/kg/day versus 16 microg/kg/day) for the steady-state mobilization of PBSCs. The effects of a filgrastim dose of 8 microg/kg/day were not significantly different from those of a dose of 16 microg/kg/day. In the group of patients receiving 8 microg/kg/day, the CD34+ cells over 3 x 10(6)/kg donor body weight were harvested in 3 patients who did not have a long history of receiving combination chemotherapy. The administration of 8 microg/kg filgrastim was adopted also for allogeneic PBSC mobilization for 24 healthy donors. All healthy donors donated an adequate number of PBSCs (CD34+ cells over 4 x 10(6)/kg of recipient body weight) and tolerated this mobilization well with no serious complications. In PBSC mobilization with healthy donors, the maximal yields of CD34+ cells from Day 4 to Day 6 were seen on the fifth day in most cases.