儿童外周血造血干细胞采集影响因素分析

目的探讨性别、年龄、外周血白细胞(WBC)和单个核细胞(mononuclear cell,MNC)计数对儿童外周血造血干细胞采集效果的影响。方法收集2016年1月~2018年1月期间,本院采集的儿童外周血干细胞28例,年龄2岁10个月~14岁,中位年龄10岁。共采集31次,自体7例次采用化疗+粒系集落刺激因子动员,异体24例次采用粒系集落刺激因子动员,均在动员后4~5 d进行外周血干细胞采集。单次平均循环血量(7593±2248)ml,单次产品体积(126±42)ml。干细胞产品进行血常规计数和流式细胞仪检测CD34~+细胞数量。结果儿童外周血干细胞单次采集产品MNC与性别、年龄无关(P=0.44;P=0.81),与外周血WBC、MNC计数相关(P=0.03;P=0.01),产品CD34~+细胞数与性别、年龄、外周血WBC无关(P=0.15;P=0.79;P=0.66),与采前MNC计数相关(P=0.02)。其中24例患者采集前外周血MNC3.75×10~9/L,单次采集的干细胞产品MNC数为(9.88±3.16)×10~8/kg,CD34+细胞数为(7.51±4.74)×10~6/kg。结论外周血MNC数量是儿童外周血造血干细胞采集效率的重要影响因素,动员后外周血MNC数量3.75×10~9/L,具有做为造血干细胞动员效果评价指标的应用价值。     

未动员的外周血造血干细胞采集效果及影响因素

目的探讨未动员的外周血造血干细胞(PBSC)采集效果及影响因素。方法应用血细胞分离机对112例未经重组人粒细胞集落刺激因子(rhG-CSF)动员的健康供者进行PBSC采集,并分析年龄、体质量指数(BMI)、采集前血常规指标、采集循环血容量、处理血量及循环次数等因素对男女两组供者所采集获得的单个核细胞(MNC)、CD34+计数的影响,同时比较分析男女两组供者采集前血常规指标及采集过程、采集物等指标。结果男性组年龄,BMI,采集前血细胞比容(Hct)、血红蛋白(Hb)、MNC计数、白细胞(WBC)计数,总循环血量高于女性组,而采集处理血量、采集物中MNC计数低于女性组,差异均有统计学意义(P<0.05)。在男性组中,采集物中MNC计数的影响因素为采集循环数(P=0.018),CD34+计数的影响因素为采集前血小板(PLT)计数(P=0.048)。女性组年龄、采集前PLT计数、WBC计数、MNC百分比、MNC计数是采集物中MNC计数的影响因素(P<0.05),采集前Hct、PLT计数及采集处理血量是采集物中CD34+计数的影响因素(P<0.05)。结论未经rhG-CSF动员的健康供者其外周血中存在一定数量的MNC、CD34+细胞,并能采集到满足临床嵌合抗原受体T细胞疗法所需要的MNC阈值。健康供者不同年龄、性别、总循环血量等可致采集效果不一致;采集前关注血常规中PLT计数有助于预测PBSC采集效果。     

无关供者24例外周血干细胞动员和采集

目的 总结无关供者外周血千细胞(PBSC)动员和采集情况.方法 24例无关供者给予重组人粒细胞集落刺激因子(rhG-CSF)5 μμg·kg-1·d-1,每天皮下注射,第4、5天或5、6天用CS-3000PLUS血细胞分离机采集外周血干细胞,计数采集物中单个核细胞(MNC)和CD34+细胞.结果 所有供者均安全顺利完成...     

外周血CD34+细胞计数预测造血干细胞收获量

目的了解外周血CD34+细胞计数与造血干细胞收获量的关系.方法用流式细胞仪Pro-COUNT方法检测了16例造血干细胞移植患者单采前外周血和采集物中CD34+细胞百分率、绝对数,用血细胞计数仪计数白细胞数.分析各项检测指标与CD34+细胞收获量(×106/kg体重)的相关性.结果外周血白细胞计数、CD34+百分率、CD34+细胞计数与CD34+细胞收获量的相关系数分别为-0.307,0.738,0.665.经相关系数t检验,单采前外周血白细胞计数与CD34+收获量无相关性(P＞0.05),而外周血CD34+百分率和绝对计数与CD34+收获量密切相关(P＜0.01).结论外周血CD34+计数可准确预测采集效果,确定干细胞采集时机.     

外周血干细胞移植中标本放置时间对CD34~+细胞及单个核细胞计数结果的影响

目的探讨标本放置温度及时间对外周血CD34+细胞及单个核细胞(MNC)计数结果的影响。方法抽取10例健康供者或自体外周血干细胞(APBSCT)移者经粒细胞集落刺激因子(G-CSF)动员第5天EDTA-K2抗凝静脉血,各分装2管,分别在室温和4℃条件下保存,对每份外周血标本分别于0、1、2、4、6、8、10、12、24h计数MNC和CD34+细胞。结果室温放置的标本中,MNC和CD34+细胞计数随着标本放置时间的延长而逐渐减低,放置到8h时,CD34+细胞计数和0h相比差异有统计学意义(t=5.04,P<0.05);放置到12h时,MNC计数和0h相比差异有统计学意义(t=3.68,P<0.05)。4℃条件下,标本放置到24h,MNC和CD34+细胞计数结果和0h相比差异均无统计学意义(t分别为0.50、1.24,P>0.05)。结论为确保检测结果的准确性,室温放置的标本CD34+细胞计数应在采血后8h内完成,MNC计数可在采血后10h内完成,外周血标本最好放置于4℃条件下保存。     

肿瘤患者自体外周血干细胞采集效果与影响因素的研究

目的探讨影响肿瘤患者行外周血干细胞采集效果的因素。方法采用MCS+血细胞分离机对79例采集前未动员的肿瘤患者(A组)行自体血外周血干细胞(PBSC)采集,所采集获得单个核细胞(MNC)和CD34~+细胞数分别与采集前白细胞(WBC)计数、血红蛋白(Hb)、体质量指数(BMI)、甘油三酯(TG)、载脂蛋白B(apoB)、红细胞比容(Hct)、血小板计数等因素进行相关性分析;同时与52例采集前动员的血液肿瘤患者(B组)所采集获得MNC和CD34~+细胞数进行比较。结果未动员的A组患者均采集成功:(1)其外周血中存在并能够采集到一定数量的MNC、CD34~+细胞。(2)与B组(动员组)比较,A组MNC、CD34~+细胞总数明显低于B组,差异有统计学意义(P<0.01),但A组与B组的MNC和CD34的百分数比较差异无统计学意义(P>0.01)。(3)采集物的MNC、CD34~+细胞数量与采集前自身的WBC计数、Hb、BMI、apoB、Hct、血小板计数、MNC均无明显相关性,而采集的MNC细胞数与采集前自身的TG水平是呈负相关。结论未作动员的肿瘤患者其外周血都可以安全地采集到一定浓度的、并能够满足临床治疗需要的PBSC。肿瘤患者自身PBSC采集效果与采集前TG水平因素有关联。     

rhIL-11联合rhG-CSF动员小鼠外周血造血干/祖细胞的研究

目的　研究rhIL 11对小鼠巨核系造血干 /祖细胞的动员作用。方法　rhIL 112 5 0μg·kg-1·d-1或联合rhG CSF 2 5 0 μg·kg-1·d-1给C5 7BL/ 6小鼠皮下注射 1～ 7d ,观察用药前和用药第 3,5 ,7,9天小鼠外周血白细胞、血小板计数 ,CD34 +细胞比例 ,CFU GM、CFU MK、CFU E的数量变化。结果　单用rhIL 11或与rhG CSF联合使用时 ,外周血白细胞、血小板、CD34 +细胞比例及各种造血细胞集落数明显高于对照组 (P <0 .0 1)。在含有IL 11的实验组中 ,CFU MK明显高于rhG CSF组 (P <0 .0 1)。结论　rhIL 11可升高外周血白细胞、血小板 ,同时增加外周血CD34 +细胞的比例 ,提高粒、红、巨核系造血细胞集落形成单位的数量 ,特别是对CFU MK作用较强 ;与rhG CSF联合使用对动员骨髓造血干 /祖细胞进入外周血有明显的协同作用。     

血细胞分离机采集干细胞效果观察

目的探讨Amicus血细胞分离机的MNC程序采集外周血干细胞的效果。方法用Amicus血细胞分离机的MNC程序采集健康供者和肿瘤患者外周血干细胞;供者给予G-CSF动员,患者采用化疗加G-CSF动员。用流式细胞仪检测CD34~+抗原表达细胞数。供患者共采集了53次,处理(8±2)个循环,处理抗凝全血(11420±2401)ml,时间(236±28)min,抗凝剂(957±195)ml。结果采集CD34+细胞(235.26±298.53)×10~6,MNC的采集效率为(53.05±39.03)%;患/供者采集前外周血CD34~+计数>0.04×10~9/L(n=28),采集CD34~+细胞为(4.94±4.57)×10~6/kg;而当患/供者采集前外周血CD34~+计数<0.04×10~9/L(n=25),采集CD34~+细胞为(1.07±0.64)×10~6/kg;所采集的干细胞制品中血小板含量为(5.57±4.26)×10~(10)/袋。单采后患/供者Plt、Hb、Hct分别下降14.79%,12.21%和12.23%,所有程序没有观察到严重的副反应。结论Amicus血细胞分离机的MNC采集程序能安全地采集到血小板含量低、高产量的异体和自体CD34~+细胞。     

血缘关系供者外周血干细胞移植中移植物细胞组分对造血重建移植物抗宿主病的影响

目的 探讨HLA全相合血缘关系供者外周血干细胞移植(PBSCT)中移植物细胞组分对恶性血液病患者移植后造血重建、移植物抗宿主病(GVHD)的影响.方法 回顾性分析我科107例接受HLA全相合血缘关系供者PBSCT的恶性血液病患者,其移植物细胞组分与移植后患者造血重建、GVHD的关系.结果 移植物各细胞组分与粒细胞重建时间无关;单个核细胞(MNC)、CD34+细胞数与血小板重建时间呈负相关(r值分别为-0.32和-0.21,P值均＜0.05).CD34+、CD34+CD38-细胞数与急性GVHD发生呈负相关(r分别为-0.24和-0.29,P值均＜0.05).淋巴细胞各亚群数量与急性GVHD发生均无明显关系.CD25+ CD4+、CD3+、CD4+ CD3+细胞数及CD4+/CD8+细胞比值与慢性GVHD发生均呈正相关(P值均＜0.05),且相关系数均大于0.4,其中CD25+ CD4+细胞数与慢性GVHD相关系数高达0.78.CD34+、CD34+ CD38-细胞数与慢性GVHD发生无明显关系.结论PBSCT中输入MNC、CD34+、CD34+ CD38-细胞数增加到一定阈值后,增加细胞数并不能进一步有效促进患者造血重建,反而有可能因输入淋巴细胞数增加而增加患者慢性GVHD、广泛慢性GVHD的发生率.     

未动员外周血造血干细胞采集的临床效果评价

目的探讨未动员外周血造血干细胞(PBSC)采集影响因素及采集效果。方法应用MCS+血细胞分离机对该院130例未动员采集者进行PBSC采集,获得单个核细胞(MNC)、白细胞(WBC)、CD34+细胞数分别与采集前体质量指数(BMI)、三酰甘油(TG)、载脂蛋白(apo)B、红细胞比容(Hct)、血小板(PLT)、WBC、血红蛋白(Hb)、MNC等指标,以及采集时循环数、循环血量等因素进行相关性分析;同时对其中50岁以下的2组(健康组与疾病组)的PBSC采集效果进行比较。结果所有采集者均采集成功:(1)其外周血中是存在并能够采集到一定数量的MNC、CD34+细胞的。(2)50岁以下采集者比较:健康组采集物中MNC、WBC计数均高于疾病组,差异有统计学意义(P0.05);但2组采集物CD34+细胞数比较差异无统计学意义(P0.05)。(3)采集物的MNC总量和CD34+细胞数量与采集前自身的apoB、PLT、WBC、Hb、TG均无明显相关性。采集前的外周血MNC、BMI、Hct及采集时循环血量、循环数对采集效果有相关影响,其中外周血MNC水平与采集物MNC水平呈正相关。结论未作动员的采集者其外周血可安全采集到一定浓度的、并能够满足临床治疗需要的PBSC。采集者自身PBSC采集效果与采集前MNC、BMI、Hct水平及采集时循环血量、循环数有关联,其中采集前MNC数量可预测采集者PBSC采集效果。     

The absolute number of circulating CD34+ cells as the best predictor of peripheral hematopoietic stem cell yield.

Many controversies still exist about the timing of leukapheresis procedures for PBSC transplantation. Thirty-nine patients were followed daily by monitoring the absolute PB WBC count and CD34+ cell enumeration prior to apheresis. These determinations were compared with the apheresis cell content (nucleated cells and CD34+ cells yield). There was a highly significant correlation between PB CD34+ cells and apheresis CD34+ cell yield (r = 0.921, p < 0.001). A small but significant correlation was found between the PB WBC count and the apheresis nucleated cell content (r = 0.383, p < 0.001), but no correlation was found between PB WBC count and apheresis CD34+ cell yield (r = -0.065, p = 0.460). A target value of 20 x 10(6) CD34+ cells/L was determined to be the most reliable predictor to collect at least 1.0 x 10(6) CD34+ cells/kg in a single apheresis. Of the 39 patients, 20 could be followed after transplantation, and a good correlation was found for total number of CD34+ cells reinfused and platelet and neutrophil engraftment. No correlation was found for nucleated cells infused and engraftment. CD34+ cell determination is a better predictor than WBC count for timing leukapheresis and is thus recommended for monitoring the quality of the product.     

Evaluation of an algorithm based on peripheral blood hematopoietic progenitor cell and CD34+ cell concentrations to optimize peripheral blood progenitor cell collection by apheresis

BACKGROUND: Quantification of peripheral blood (PB) CD34+ cells is commonly used to plan peripheral blood progenitor cell (PBPC) collection but is time-consuming. Sysmex has developed a hematology analyzer that can quickly identify a population of immature hematopoietic cells (HPCs) according to cell size, cell density, and differential lysis resistance, which may indicate the presence of PBPCs in PB. This prospective study has evaluated the potential of such method to predict the PBPC mobilization. STUDY DESIGN AND METHODS: A total of 141 patients underwent PBPC mobilization. PB HPCs and PB CD34+ cells were simultaneously quantified with a hematology analyzer (SE2100, Sysmex) and flow cytometry, respectively. The number of blood volumes processed was then based on PB CD34+ cell concentration. RESULTS: The optimal PB HPC level able to predict a minimal level of 10 x 10(6) PB CD34+ cells per L was 5 x 10(6) per L with positive and negative predictive values of 0.93 and 0.36 percent, respectively. For this cutoff point, sensitivity and specificity were 0.81 and 0.65, respectively. The median number of blood volumes processed according to the PB CD34+ cell count allowed us to perform only one apheresis procedure for a majority of patients. CONCLUSION: PB HPC quantification is very useful to quickly determine the initiation of PBPC apheresis especially for patients with higher concentrations. For patients exhibiting a lower HPC count (<5 x 10(6)/L), other parameters such as a CD34 test may be needed. Such a policy associated with a length of apheresis adapted to the richness in the PB CD34+ cells allows for optimizing the organization of centers with an improvement in patient comfort and economical savings.     

The predictive value of white cell or CD34+ cell count in the peripheral blood for timing apheresis and maximizing yield

BACKGROUND: The collection of peripheral blood stem and progenitor cells (PBPCs) for transplantation can be time-consuming and expensive. Thus, the utility of counting CD34+ cells and white cells (WBCs) in the peripheral blood was evaluated as a predictor of CD34+ cell yield in the apheresis component. STUDY DESIGN AND METHODS: The WBC and CD34+ cell counts in the peripheral blood and the apheresis components from 216 collections were assessed. Sixty-three patients underwent mobilization with chemotherapy plus filgrastim, and 17 patients and 14 allogeneic PBPC donors did so with filgrastim alone. The relationship between the number of WBC and CD34+ cells in the peripheral blood and in the apheresis component was analyzed by using rank correlation and linear regression analysis. RESULTS: The correlation coefficient for CD34+ cells per liter of peripheral blood with CD34+ cell yield (x 10(6)/kg) was 0.87 (n = 216 collections). This correlation existed for many patient and collection variables. However, patients with acute myeloid leukemia had fewer CD34+ cells in the apheresis component at any level of peripheral blood CD34+ cell count. Components collected from patients with CD34+ cell counts below 10 x 10(6) per L in the peripheral blood contained a median of 0.75 x 10(6) CD34+ cells per kg. When the WBC count in the blood was below 5.0 x 10(9) per L, the median number of CD34+ cells in the peripheral blood was 5.6 x 10(6) per L (range, 1.0-15.5 x 10(6)/L). A very poor correlation was found between the WBC count in the blood and the CD34+ cell yield (p = 0.12, n = 158 collections). CONCLUSION: The number of CD34+ cells, but not WBCs, in the peripheral blood can be used as a predictor for timing of apheresis and estimating PBPC yield. This is a robust relationship not affected by a variety of patient and collection factors except the diagnosis of acute myeloid leukemia. Patients who undergo mobilization with chemotherapy and filgrastim also should undergo monitoring of peripheral blood CD34+ cell counts, beginning when the WBC count in the blood exceeds 1.0 to 5.0 x 10(9) per L.     

Peripheral blood circulating immature cell counts predict CD34+ cell yields in G-CSF-induced PBPC mobilization in healthy donors

BACKGROUND: It has been previously reported that the number of circulating immature cells (CIC) in peripheral blood (PB) estimates the number of CD34+ cells collected in G-CSF plus chemotherapy-induced PBPC mobilization. The correlation of CIC counts in PB with CD34+ cell yield and its usefulness was evaluated in G-CSF-induced PBPC mobilization for healthy donors. STUDY DESIGN AND METHODS: CIC counts in PB and CD34+ cell counts in the apheresis product from 122 collections were assessed, and the relationship between these two variables was evaluated with the Pearson rank correlation analysis, the chi-squared test, and the U-test. RESULTS: CIC counts were correlated weakly with the number of CD34+ cells per L of blood processed in the apheresis product (Pearson rank correlation analysis; r=0.357, p<0.0001). When a level of 1.7 x 10(9) CICs per L was selected as a cutoff value, the sensitivity and specificity for collecting more than 20 x 10(6) CD34+ cells per L of blood processed were 63.6 and 77.5 percent, respectively. CONCLUSION: The present study suggests that the number of CICs in PB may estimate the number of CD34+ cells collected. The data indicate that CIC counts above 1.7 x 10(9) per L can be used as a good predictor for PBPC collections containing more than 20 x 10(6) CD34+ cells per L of blood processed in a single apheresis procedure.     

Collection efficiencies of MNC subpopulations during autologous CD34+ peripheral blood progenitor cell (PBPC) harvests in small children and adolescents

There is increasing demand for mononuclear cell (MNC) harvests not only for PBPC but also for immune therapies using dendritic cells and donor lymphocytes. We determined the collection efficiencies (CE) of various MNC subpopulations during CD34+ cell harvests using a Fenwal CS 3000 Plus Omnix system in small children and adolescents. The cell content of 140 leukapheresis products (LP) was prospectively evaluated in 45 pretreated patients with solid tumors and hematological malignancies. The median age was 12 years (range 0.8-22), and the median body weight (BW) 43 kg (range 9-92). Depending upon the BW of the patients, the media used for priming were saline (SP) in 86, human albumin (HA, HAP) in 10, and packed red blood cells (BP) in 44 apheresis procedures. The major nucleated cell (NC) fractions collected were monocytes (52% of NC) and CD3+ T cells (26%). The median cell yield for monocytes was 174 * 10(6)/kg (range 24-613) representing a CE of 55%. The median number of CD3(+) T cells was 84 * 10(6)/kg (range 5.6-380; CE = 74%). CD34+ cells represented a very small cell fraction of the LP (1.3% of NC), with a median yield of 4.2 * 10(6)/kg (range 0.2-87) and a CE of 63%. The cell yield of various MNCs was significantly correlated with the cell count in the peripheral blood (PB) and with the blood volume processed (ANOVA, P < 0.0001). No influence on the CE was observed for the priming procedure, the patients' age or sex, or the other adaptations used in the harvesting protocol. In conclusion, the Fenwal CS 3000 Plus OMNIX system with the CD34+ cell program and the described adaptations, is also predictably useful for harvest of monocytes or lymphocytes in pediatric patients. We present regression equations that predict the cell yield of various MNC subpopulations in apheresis products.     

Monitoring of peripheral blood CD34+ cell counts on the first day of apheresis is highly predictive for efficient CD34+ cell yield.

The purpose of this study was to evaluate the correlation of preleukapheresis circulating CD 34+ cells/micro L, white blood cells (WBC), and platelet counts on the first day of apheresis with the yield of collected CD 34+ cell counts in 40 patients with hematological malignancies (n = 29) and solid tumors (n = 11). The median numbers of apheresis cycles, numbers of CD 34+ cells, peripheral blood (PB) mononuclear cells, and total nucleated cells collected were 2 (range, 1-4), 5.5 x 106/kg (range, 0.05-33.78), 2.59 x 108/kg (range, 0.04-20.68), and 7.36 x 108/kg (range, 0.15-28.08), respectively. There was a strong correlation between the number of preleukapheresis circulating CD 34+ cells/micro L and the yield of collected CD 34+ cells per kilogram (r = 0.962, p < 0.001). The threshold levels of PB C 34+ cell/micro L to obtain > or =1 x 106/kg and > or =2.5 x 106/kg CD 34+ cell in one collection were 12/micro L and 34/ micro L, respectively. Fifteen of 17 (88%) patients who had > or =34 CD 34+ cells/ micro L in the PB before collection reached the level of > or =2.5 x 106/kg in a single apheresis. Despite a low r value, WBC and platelet counts on the first day of apheresis also correlated with the yield of collected daily CD 34+ cells per kilogram (r = 0.482, p < 0.01 and r = 0.496 p < 0.01, respectively). These data suggest that preleukapheresis circulating CD 34+ cells/ micro L correlated significantly better with the yield of collected CD 34+ cells than WBC and platelet counts on the first day of apheresis. Using a value of 34/micro L preleukapheresis circulating CD 34+ cells as a guide for the timing of peripheral blood stem cells collections can be time saving and cost-effective.     

Evaluation of a new cell separator for collection of peripheral blood CD34+ progenitor cells in pediatric patients

BACKGROUND: This study was conducted to evaluate the performance of the COM.TEC cell separator (Fresenius HemoCare GmbH) for collecting CD34+ cells in pediatric patients who were intended to have autologous peripheral blood progenitor cell transplantation, with respect to collection variables, prediction power of CD34+ cell yield, and influence on donors. STUDY DESIGN AND METHODS: A total of 26 pediatric solid tumor patients who received mobilization chemotherapy and granulocyte–colony‐stimulating factor underwent CD34+ cell collection (n = 96) using the COM.TEC auto mononuclear cell (MNC) program. Patients were divided into a neuroblastoma (NBL) group and a brain tumor group according to the intensity of prior chemotherapy regimens. The collection variables, cellular variables of leukapheresis products, and the peripheral blood cell counts of patients were compared with those acquired using the COBE Spectra (GambroBCT). The CD34+ cell collection efficiency (CE) and the percentage ratios of actual to predicted CD34+ cell yield indicating prediction power were analyzed. RESULTS: Using the COM.TEC auto MNC program, the processing rate was higher and the product volume was smaller (p < 0.05) than those of the COBE Spectra. Platelet (PLT) reduction in peripheral blood and PLT contamination of the products were significantly lower (p < 0.01). The median CE was less than 60% in both patient groups (50.0 and 48.4%, respectively). The actual collected CD34+ cell yields were medians of 66.9 and 76.1% of the predicted values in NBL group and brain tumor group, respectively. CONCLUSION: PBPC collections by the COM.TEC cell separator had advantages of high processing rate, low product volume, and low contamination by PLTs of product. Low PLT loss was observed in pediatric patients who need to collect autologous PBPCs. However, applying CD34+ cell yield prediction was not practical for prospective scheduling of the next collection. More specified data need to be accumulated for more accurate prediction of CD34+ cell yield in pediatric patients.     

Immune reconstitution after autologous selected peripheral blood progenitor cell transplantation: comparison of two CD34+ cell-selection systems

BACKGROUND: Selection of CD34+ PBPCs has been applied as a method of reducing graft contamination from neoplastic cells. This procedure seems to delay lymphocyte recovery, while myeloid engraftment is no different from that with unselected PBPC transplants. STUDY DESIGN AND METHODS: Lymphocyte recovery was studied in two groups of patients who underwent autologous CD34+ PBPC transplant with two different technologies (Ceprate SC, Cellpro [n = 17]; CliniMACS, Miltenyi Biotech [n = 13]). The median number of CD34+ cells transfused was 3.88 x 10(6) per kg and 3.32 x 10(6) per kg, respectively. Residual CD3 cells x 10(6) per kg were 4.97 and 0.58, respectively (p = 0.041). Residual CD19 cells x 10(6) per kg were 1.33 and 0.73, respectively (NS). RESULTS: No differences were found between the two groups in total lymphocyte recovery to >0.5 x 10(9) per L, which achieved a stable count by Day 30. During the study period, the CD4+ cell count remained below 0.2 x 10(9) per L, and the B-cell subset showed a trend toward normalization. CD3/HLA-DR+ and CD16/56 increased markedly in both groups by Day 30. An increase in CMV (13%) and adenovirus (17.4%) infection was found in both groups. CONCLUSION: Both CD34+ cell selection technologies used here determined an excellent CD34+ cell purity and an optimal depletion of T cells. The high rate of viral complications is probably due to the inability of residual T cells left from the CD34+ cell selection to generate, immediately after transplant, an adequate number of virus-specific lymphocytes.     

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.     

Comparison of COBE® Spectra™ software version 4.7 PBSC and version 6.0 auto PBSC™ program

Until recently, the collection of peripheral blood progenitor cells (PBPC) has been semi-automated by using the COBE® Spectra™, with the operator manually maintaining the position of the white cells being collected. The COBE® Spectra™ Version 6.0 apheresis device offers the user an automated program for the collection of PBPC. In this study, we compared the new software Version 6.0 to that of Version 4.7. Patients (n=46) undergoing PBPC collection were allocated to cell processing with either Version 4.7 (n=24) or Version 6.0 (n=22). The CD34+ cell count, mononuclear cell (MNC) count, white cell count (WCC), hemoglobin (Hb), and platelet content in the autograft product by using the two versions were compared. We divided the analysis into three subsets according to peripheral blood (PB) CD34 content: <10×10⁶/L, 10–50×10⁶/L and >50×10⁶/L. Analysis of the three subsets showed no statistical difference between results obtained when the starting PB CD34+ cell count was 10–50×10⁶/L (P=0.08) or >50 ×10⁶/L (P=0.4065). At lower starting PB CD34+ cell counts of <10×10⁶/L, Version 4.7 was superior (P=0.0167). However, autograft platelet contamination of the autograft was significantly higher using Version 4.7 (P=<0.0001). J. Clin. Apheresis 14:26–30, 1999. © 1999 Wiley-Liss, Inc.