Blood graft lymphocyte subsets after plerixafor injection in non-Hodgkin's lymphoma patients mobilizing poorly with chemotherapy plus granulocyte-colony-stimulating factor

BACKGROUND: A combination of chemotherapy plus granulocyte–colony‐stimulating factor (G‐CSF) (chemomobilization) is commonly used to mobilize CD34+ cells to circulation. Plerixafor, a chemokine CXCR4 antagonist, increases the mobilization of CD34+ cells and may also have effect on graft composition. STUDY DESIGN AND METHODS: We have analyzed lymphocyte subsets in grafts collected on the next morning after plerixafor injection in 13 chemomobilized patients with non‐Hodgkin's lymphoma (NHL) mobilizing poorly. As controls we had the first leukapheresis products from 11 NHL patients mobilized with chemotherapy plus G‐CSF. The analyses were performed from cryopreserved apheresis products. RESULTS: The median counts of both total CD3+ T cells and natural killer (NK) cells (CD3–CD16/56+) in the graft were significantly higher in plerixafor‐treated group compared to the control group. Both helper T‐lymphocytes (CD3+CD4+) and suppressor T‐lymphocytes (CD3+CD8+) were significantly increased in the plerixafor‐treated group so that CD4+/CD8+ ratio in the graft did not differ between the groups. CD19+ cells were evident only at very small amounts in few patients in both groups, and the CD34+ cell content of the graft did not differ between the groups. Engraftment after high‐dose therapy was comparable between the groups. CONCLUSION: Plerixafor added to chemomobilization in poor mobilizers seems to mobilize more T cells and NK cells than chemomobilization. Larger patient numbers and longer follow‐up is needed in regard to evaluate posttransplant complications and risk of relapse in patients receiving plerixafor due to poor mobilization.     

Mobilization of blood-derived stem and progenitor cells in normal subjects by granulocyte-macrophage- and granulocyte-colony-stimulating factors

BACKGROUND: It was previously reported that the combination of granulocyte-macrophage-colony-stimulating factor (GM-CSF) and granulocyte-CSF (G-CSF) for 4 days mobilized more primitive CD34+ subsets than did either G-CSF or GM-CSF alone. STUDY DESIGN AND METHODS: The studies determine the optimal number of days of growth factor dosing for mobilization and collection of peripheral blood progenitor cells, by increasing the days of administration of GM-CSF and/or G-CSF or employing the sequential administration of GM-CSF followed by G-CSF. Sixty normal subjects were given injections of G-CSF or GM-CSF alone; GM-CSF and G-CSF concurrently for 4, 5, or 6 days; or a sequential regimen of GM-CSF for 3 or 4 days followed by G-CSF for 2 or 3 days. A 10-L apheresis was performed 24 hours after the last dose. RESULTS: The three most efficacious mobilization regimens consisted of sequential GM-CSF for 3 days followed by G-CSF for either 2 or 3 days and G-CSF alone for 5 days. Each of these regimens resulted in the collection of significantly greater numbers of CD34+ cells by apheresis than any of the 4-day dosing regimens with G-CSF and/or GM-CSF (sequential GM-CSF/G-CSF: 3 days/2 days = 3.58 +/− 0.53 × 106 CD34+ cells/kg; GM-CSF/G-CSF: 3 days/3 days = 4.45 +/− 1.08 × 10(6) CD34+ cells/kg; G-CSF: 5 days = 3.58 +/− 0.97 × 10(6) CD34+ cells/kg; all p<0.05 vs. G-CSF and/or GM-CSF for 4 days). Clonogenic assays generally paralleled the level of CD34+ cells. Regimens containing GM-CSF resulted in a higher percentage of the cells from primitive CD34+/CD38- /HLA-DR+ subset than G-CSF alone. CONCLUSION: Compared with 4-day dosing regimens with G-CSF and/or GM-CSF, mobilization of CD34+ cells in normal subjects using sequential GM-CSF for 3 days followed by G-CSF for 2 or 3 days or using G-CSF alone for 5 days increased the number CD34+ cells that can be collected by a single 10-L apheresis 24 hours after the last dose of cytokine.     

正常人骨髓、脐血及动员后外周血CD^＋34细胞粘附分子的研究

目的探讨正常人骨髓、脐血及动员后外周血CD34+细胞粘附分子的表达及外周血干细胞动员的可能机制.方法采用CD34+MultiSortKit免疫磁珠分离系统,分离纯化出正常人骨髓、脐血及动员后外周血CD34+细胞,流式细胞术检测其纯度,选择与CD34+细胞相关的粘附分子CD44、CD11a、CD18、CD49d、CD54、CD58及CD62L,进行免疫荧光标记及短期液体培养后再行免疫荧光标记,流式细胞术检测.结果动员后外周血CD34+细胞粘附分子表达CD44为(92.7±2.2)%[骨髓(93.1±2.3)%]、CD11a为(56.3±6.0)%[骨髓(61.8±7.8)%]、CD18为(65.2±6.0)%[骨髓(70.6±7.5)%]、CD49d为(39.4±7.2)%[骨髓(66.9±5.1)%]、CD54为(20.9±4.1)%[骨髓(24.1±3.8)%]、CD58为(77.9±5.8)%[骨髓(81.9±5.6)%]及CD62L为(45.9±5.6)%[骨髓(63.9±4.3)%],其表达均较骨髓为低,尤以CD49d和CD62L为著.脐血CD34+细胞CD11a为(55.5±6.5)%、CD18为(66.7±7.5)%、CD44为(90.3±4.0)%、CD49d为(63.7±6.7)%、CD62L为(50.8±5.9)%,其表达亦较骨髓为低,尤以CD62L为著,但脐血CD54的表达[(29.1±4.9)%]较骨髓及动员后外周血为高,尤较动员后外周血为著.结论不同来源CD34+细胞粘附分子表达存在差异,外周血细胞动员的机制可能与粘附分子的表达下调有关.     

Management strategies for poor peripheral blood stem cell mobilization.

Peripheral blood stem cells (PBSC) have nearly replaced bone marrow (BM) as the preferred source of hematopoietic rescue for patients undergoing high-dose chemotherapy. However, some patients fail to mobilize sufficient numbers of PBSC into the peripheral blood thereby putting high-dose chemotherapy at risk. The present article reviews mobilization of PBSC with a special focus on poor mobilizers. Under steady-state conditions less than 0.05% of the white blood cells (WBC) are CD34+ cells. Chemotherapy results in a 5-15-fold increase of PBSC. Combining chemotherapy and growth factors increases CD34+ cells up to 6% of WBC. Several factors affect the mobilization of PBSC: age, gender, type of growth factor, dose of the growth factor and in the autologous setting patient's diagnosis, chemotherapy regimen and number of previous chemotherapy cycles or radiation. Poor mobilizers are defined as patients with less than 10 CD34+ cells/mul in the peripheral blood during mobilization. Promising approaches for those patients rely on remobilization, use of high doses of granulocyte-colony stimulating factor (G-CSF), or the combination of G-CSF and granulocyte macrophage (GM)-CSF, which successfully mobilized the majority of poor mobilizing patients. New agents such as long lasting variants of G-CSF and CXCR4 antagonists are at the horizon and studied in clinical trials as mobilizing agents. Muscle and bone pain are frequent adverse events in stem cell mobilization but are usually tolerated under the use of analgesics. Large volume apheresis (LVL) with a processed volume of more than 4-fold patient's blood volume is an approach to increase the CD34+ yield in patients with low CD34+ pre-counts resulting in higher yields of CD34+ cells for transplantation. Processing of more blood in LVL is achieved by an increase of the blood flow rate and an altered anticoagulation regimen with the occurrence of more citrate reactions.     

Augmented mobilization and collection of CD34+ hematopoietic cells from normal human volunteers stimulated with granulocyte-colony-stimulating factor by single-dose administration of AMD3100, a CXCR4 antagonist

BACKGROUND: AMD3100, a selective antagonist of CXCR4, rapidly mobilizes CD34+ hematopoietic progenitor cells (HPCs) from marrow to peripheral blood with minimal side effects. STUDY DESIGN AND METHODS: To further investigate potential clinical utility of AMD3100 for CD34+ cell mobilization and collection, a Phase I study in normal volunteers was performed examining single-dose administration of AMD3100 alone and in combination with a standard 5-day granulocyte-colony-stimulating factor (G-CSF) regimen. RESULTS: AMD3100 (160 microg/kg x 1 on Day 5) significantly increased both G-CSF-stimulated (10 microg/kg/day) mobilization of CD34+ cells (3.8-fold) and leukapheresis yield of CD34+ cells. Moreover, collection of CD34+ cells was comparable between individuals mobilized by a single-dose regimen of AMD3100 (240 microg/kg) and individuals mobilized with a 5-day regimen of G-CSF. AMD3100-mobilized leukapheresis products contained significantly greater numbers of T and B cells compared to G-CSF-stimulated leukapheresis products. CONCLUSION: These findings indicate that AMD3100 can be used alone or as an adjunct to G-CSF to mobilize cells for HPC transplantation.     

Expression of the adhesion molecules CD49d and CD49e on G-CSF-mobilized CD34+ cells of patients with solid tumors or non-Hodgkin's and Hodgkin's lymphoma and of healthy donors is inversely correlated with the amount of mobilized CD34+ cells

The yield of CD34+ PBPC and colony-forming units-granulocyte-macrophage (CFU-GM) in leukapheresis products and the expression of the adhesion molecules CD11a, CD31, CD49d, CD49e, CD54, CD58, CD62L, c-kit (CD117), Thy-1 (CD90), CD33, CD38, and HLA-DR on CD34+ PBPC were analyzed in patients with cancer of the testis (n = 10), breast cancer (n = 10), Hodgkin's disease (n = 20), high-grade (n = 20) and low-grade (n = 20) non-Hodgkin's lymphoma, and healthy donors (n = 20) undergoing G-CSF (filgrastim)-stimulated PBPC mobilization. For each disease entity, G-CSF was administered in two different doses, 10 microg G-CSF/kg body weight (BW)/day s.c. vs. 24 microg G-CSF/kg BW s.c./day in steady-state condition. Data were compared for each dose group separately. Patients with cancer of the testis and breast cancer mobilized significantly more CD34+ cells than patients with high-grade and low-grade non-Hodgkin's lymphoma and Hodgkin's disease (p<0.05). Correspondingly, expression of CD49d on CD34+ PBPC was significantly lower in the same patients with cancer of the testis compared with high-grade and low-grade non-Hodgkin's lymphoma and Hodgkins' disease and in patients with breast cancer compared with high-grade and low-grade non-Hodgkin's lymphoma, Hodgkins's disease, and healthy donors. Similar results were obtained for CD49e. These data suggest that the expression of the adhesion molecules CD49d and CD49e on G-CSF-mobilized CD34+ cells of patients with solid tumors, non-Hodgkin's lymphoma, Hodgkin's disease, and healthy donors is inversely correlated with the amount of mobilized CD34+ cells.     

Mobilization kinetics of CD34+/Thy-1dim progenitor cells during recombinant human granulocyte-colony-stimulating factor administration in normal donors

BACKGROUND: Allogeneic transplantation of granulocyte-colony- stimulating factor (G-CSF)-mobilized peripheral blood progenitor cells (PBPCs) from normal related donors is effective in achieving engraftment with a relatively short period of posttransplantation aplasia. The optimal dose and composition of PBPC transplants are unknown. The CD34+/Thy-1dim progenitor cell subset is enriched for putative stem cells. STUDY DESIGN AND METHODS: The kinetics of the primitive subpopulation were prospectively studied in nine normal donors receiving recombinant human G-CSF (6 microg/kg) subcutaneously twice daily for 6 days for collection of PBPCs for allogeneic transplantation. RESULTS: The concentration (mean +/− SD) of the circulating CD34+/Thy-1dim subset increased from a baseline of 0.9 +/− 0.9 × 10(3) to 29.2 +/− 22.1 × 10(3) per mL on Day 4 and 38.0 +/− 29.8 × 10(3) per mL on Day 6. The level of CD34+/Thy-1dim cells was closely correlated with the overall level of CD34+ cells. At baseline, CD34+/Thy-1dim cells composed 21.1 percent of the total CD34+ cells, increasing to 36.3 percent at the peak of mobilization. CONCLUSION: CD34+/Thy-1dim cells are optimally mobilized on Days 4 to 6 of recombinant human G-CSF treatment.     

Kinetics of PBPC mobilization by cyclophosphamide, as compared with that by epirubicin/paclitaxel followed by G-CSF support: implications for optimal timing of PBPC harvest

BACKGROUND: Limited information is available on the mobilization kinetics of autologous PBPCs after induction with various chemotherapy regimens. With PBPC mobilization in patients with breast cancer used as a model for chemotherapy-induced PBPC recruitment, the kinetics of progenitor cells mobilized either with cyclophosphamide (CY) or epirubicin/paclitaxel (EPI-TAX) followed by the administration of G-CSF was compared. STUDY DESIGN AND METHODS: The study included a total of 86 patients with breast cancer (stage II-IV) receiving either CY (n = 39) or EPI-TAX (n = 47), both followed by G-CSF support. The progenitor cell content in peripheral blood and apheresis components was monitored by flow cytometric enumeration of CD34+ cells. PBPC collection was started when the threshold of >20 x 10(6) CD34+ cells per L of peripheral blood was reached. RESULTS: The PBPC collection was begun a median of 9 days after the administration of EPI-TAX followed by G-CSF support, as compared to a median of 13 days after mobilization with CY plus G-CSF. After treatment with CY, the total numbers of PBPCs peaked on Day 1 of apheresis, and they rapidly declined thereafter. In contrast, treatment with EPI-TAX followed by G-CSF administration led to a steady mobilization of CD34+ cells during leukapheresis. The difference in the mobilization patterns with CY and EPI-TAX resulted in a greater yield of CD34+ cells per L of processed blood volume. Compared to EPI-TAX, mobilization with CY required the overall processing of 30 percent less whole-blood volume to reach the target yield of > or = 10 x 10(6) CD34+ cells per kg of body weight. After a median of three apheresis procedures, however, both CY+G-CSF and EPI-TAX+G-CSF were equally effective in obtaining this target yield. CONCLUSION: These results imply that specific PBPC mobilization as part of a given chemotherapy regimen should be taken into consideration before the planning of a PBPC harvest.     

A combination of low-dose cyclophosphamide and colony-stimulating factors is more cost-effective than granulocyte-colony-stimulating factors alone in mobilizing peripheral blood stem and progenitor cells

BACKGROUND: The use of peripheral blood progenitor cells (PBPCs) instead of autologous bone marrow leads to more rapid engraftment following high-dose chemotherapy. Mobilization regimens differ with respect to toxicity, efficiency, and cost. STUDY DESIGN AND METHODS: Two cohorts of patients with breast cancer received one of two mobilization regimens: granulocyte-colony-stimulating factor (G-CSF) at 10 micrograms per kg was given subcutaneously for 5 days, with leukapheresis begun on Day 6, or low-dose cyclophosphamide followed by sequential granulocyte-macrophage-CSF (GM-CSF) at 5 micrograms per kg for 5 days and by G-CSF at 10 micrograms per kg, with leukapheresis begun on Day 11. Results of CD34+ cell collection, engraftment, and costs of mobilization were determined. RESULTS: The combination chemotherapy and growth factor regimen was more efficient in mobilizing CD34+ cells. Sixty-six percent of patients reached a target 4 × 10(6) CD34+ cells per kg in a single leukapheresis session with the combination regimen, compared to 14 percent who received G-CSF alone (p < 0.01). The mean number of leukapheresis sessions required to reach a target of 4 × 10(6) CD34+ cells per kg was 1.3 for the combination regimen and 2.7 for the regimen of G-CSF alone (p < 0.01). One patient in the chemotherapy and growth factor group developed febrile neutropenia. Engraftment was similar in both cohorts of patients. The cost of mobilization, including all supplies and cryopreservation, was $7381 for the G-CSF regimen and $5508 for the chemotherapy regimen (p < 0.05). This reduction was attributed to the lower number of leukapheresis and cryopreservation sessions, which outweighed the slight increase in expense for chemotherapy and growth factor in the combination regimen. CONCLUSION: This combination mobilization regimen allowed the predictable and efficient collection of CD34+ cells from the peripheral blood in a limited number of leukapheresis sessions, which reduced the cost of mobilization by approximately 25 percent.     

Good and poor mobilizing patients differ in mobilized CD34+ cell adhesion molecule profiles

BACKGROUND: Alterations in expression of adhesion molecules are important in the trafficking of hematopoietic progenitors and probably in the mobilization process. Relatively little and conflicting data are currently available on the differences in expression between good and poor mobilizing patients. STUDY DESIGN AND METHODS: In this study, the expression of eight adhesion molecules on the collected CD34+ cells from 36 patients undergoing mobilization was determined. RESULTS: Good mobilizing patients, defined as those who collected their target in one apheresis procedure, had significantly fewer cells that expressed CD11a (LFA-1) and CD54 (ICAM-1) and borderline fewer that expressed CD11c, CD49d (VLA-4), and CD49d (VLA-5). No differences were detected in CD11b (Mac-1), CD15s (sLe(x)), or CD62L (L-selectin). Linear regression analysis identified number of prior chemotherapy courses and expression of CD11a (LFA-1) as independent predictive factors for mobilization efficiency. Good and poor mobilizing patients had approximately the same number of total CD34+ cells collected and little difference in times to engraftment. CONCLUSIONS: CD11a (LFA-1) expression inversely correlates with mobilization efficiency. Elucidation of the mechanism(s) underlying these observations will require further study.     

PBPC mobilization with paclitaxel, ifosfamide, and G-CSF with or without amifostine: results of a prospective randomized trial

BACKGROUND: The impact of amifostine on PBPC mobilization with paclitaxel and ifosfamide plus G-CSF was assessed. STUDY DESIGN AND METHODS: Forty patients with a median age of 34 years (range, 19-53) who had germ cell tumor were evaluated for high-dose chemotherapy. Patients were randomly assigned to receive either a single 500-mg dose of amifostine (Group A, n = 20) or no amifostine (Group B, n = 20) before mobilization chemotherapy with paclitaxel (175 mg/m(2)) given over 3 hours and ifosfamide (5 g/m(2)) given over 24 hours (TI) on Day 1. G-CSF at 10 microg per kg per day was given subsequent to TI with or without amifostine from Day 3 until the end of leukapheresis procedures. RESULTS: In 2 (10%) of 20 patients receiving amifostine and 3 (15%) of 20 patients not receiving it, no PBPC separation was performed because of mobilization failure. No significant differences were observed in the study arms with regard to the time from chemotherapy until first PBPC collection or the number of apheresis procedures needed to harvest more than 2.5 x 10(6) CD34+ cells per kg. Furthermore, leukapheresis procedures yielded comparable doses of CD34+ cells per kg (3.4 x 10(6) vs. 3.6 x 10(6); p = 0.82), MNCs per kg (2.7 x 10(8) vs. 2.6 x 10(8); p = 0.18), and CFU-GM per kg (15.9 x 10(4) vs. 19.3 x 10(4); p = 0.20). Patients in Group A had higher numbers of circulating CD34+ cells on Day 10 (103.0/microL vs. 46.8/microL; p = 0.10) and on Day 11 (63.0/microL vs.14.3/microL; p = 0.04) than did patients in Group B. CONCLUSION: Administration of a single dose of amifostine before chemotherapy with TI mobilized higher numbers of CD34 cells in the circulation, but did not enhance the overall collection efficiency in the present trial.     

The composition of leukapheresis products impacts on the hematopoietic recovery after autologous transplantation independently of the mobilization regimen

BACKGROUND: Effects of mobilization regimen on the composition of leukapheresis products (LPs) and on hematopoietic reconstitution after autologous peripheral blood progenitor cell transplantation (PBPCT) are not well known. STUDY DESIGN AND METHODS: The effects of three different mobilization regimens--stem cell factor (SCF) plus granulocyte colony stimulating factor (G-CSF) plus cyclophosphamide (CCP), G-CSF alone, and G-CSF plus CCP--on the composition of LPs from patients with nonhematologic PBPC malignancies compared to LPs from G-CSF-mobilized healthy donors and normal marrow (BM) samples were analyzed. The impact of LP composition on both short- and long-term engraftment after autologous PBPCT was also evaluated. RESULTS: The most effective regimen for mobilization of CD34+ hematopoietic progenitor cells (HPCs) into peripheral blood was SCF, G-CSF, and CCP, providing the highest numbers of all CD34+ HPCs subsets analyzed. Patients mobilized with SCF plus G-CSF plus CCP showed the highest numbers of neutrophils and monocytes, whereas the highest numbers of lymphocytes and NK cells were observed in LPs from G-CSF-mobilized patients. The overall number of CD34+ HPCs was the strongest factor for predicting recovery of platelets, whereas the number of myelomonocytic-committed CD34+ precursors was the most powerful independent prognostic factor for WBC and neutrophil recovery. The overall number of CD4+ T cells returned showed an independent prognostic value for predicting the occurrence of infections, during the first year after transplant. CONCLUSIONS: The use of different mobilization regimens modifies the overall number of CD34+ HPCs obtained during leukapheresis procedures, and also affects both the absolute and the relative composition of the LPs in different CD34+ and CD34- cell subsets.     

Mobilization of peripheral blood stem cells for autologous transplantation patients with hematological malignancies: Influence of disease, mobilization method, age and sex

Autologous peripheral blood stem cells transplantation (Auto-PBSCT) is a therapeutic option which can be used in various hematological neoplastic disorders; and it can prolong disease-free survival and total survival. Many factors could influence the mobilization of peripheral blood stem cells for patients of Auto-PBSCT. In this study, we investigated the variables influencing the mobilization of peripheral blood stem cells in 240 patients with hematological malignancies who had undergone Auto-PBSCT between 2001 and March 2007 in our center, retrospectively. Patients with acute myelogenous leukemia had the most collected mononuclear cells (MNCs) and patients with acute lymphoblastic leukemia had the most collected CD34(+) cells than did other patients. However, patients with multiple myeloma had the least collected MNCs and CD34(+) cells. Patients mobilized with chemotherapy with granulocyte colony stimulating factor (G-CSF) plus recombinant human interleukin-11(rhIL-11) had the most collected MNCs and CD34(+) cells. The difference is statistical signification between chemotherapy with G-CSF and chemotherapy with G-CSF plus rhIL-11 for collected MNCs (P<0.05). Adults had the most collected MNCs and CD34(+) cells and the difference is statistical signification between children/adolescent and older, children/adolescent and adult for CD34(+) cells (P<0.05). Male patients had the more collected MNCs and CD34(+) cells and the difference is statistical signification for CD34(+) cells (P<0.05). The adverse events were not serious during mobilization. In conclusion, many factors could influence the mobilization of peripheral blood stem cells, and our findings emphasize the need to optimize harvesting technique to enhance safety and minimize morbidity and costs of this valuable procedure.     

恶性血液系统疾病自体外周血造血干细胞动员的临床分析

目的:分析恶性血液系统疾病患者外周血造血干细胞动员与采集过程中的影响因素。方法:对50例血液系统恶性疾病患者在东南大学附属中大医院血液科进行外周血造血干细胞动员。对患者年龄、性别、动员方案、疾病状态、采集机器等因素进行分析,评估以上因素对干细胞动员结果的影响,并分析了采集前白细胞、血红蛋白、血小板的数量与采集的CD34^+细胞计数的相关性。结果:动员方案对CD34^+细胞采集数及CD34^+细胞采集成功率的影响有显著性影响,而性别、年龄、确诊到动员间隔时间、既往化疗方案、骨髓受累与否等对干细胞采集数量影响并不显著。采集前外周血白细胞数量及血红蛋白数量与采集的CD34^+细胞数呈正相关。采集前外周血中白细胞计数及单个核细胞计数与采集成功密切相关。结论:化疗联合细胞因子的动员方案采集造血干细胞优于单用细胞因子的动员方案。通过采集前白细胞计数及单个核细胞计数确定合适的采集时机,可以提高采集的成功率。     

化疗联合G-CSF动员的骨髓及外周血CD34+细胞表达粘附分子的变化

探讨化疗合G－CSF动员前，后患骺髓及外周血CD34^ 细胞表达CD44，CD49d，CD62L及趋化因子CXCR4的动态变化，用免疫荧光直接三角标记和流式细胞术测定G－CSF动员前，后骨髓及外周血CD34^ 细胞的CD44，CD49d，CD62L及趋化因子CXCR4的表达，观察输注各表达亚群细胞数与移植后造血重建的关系。结果发现，动员后骨髓中CD34^ CD44^ 和CD34^＋CD49d^ 细胞的百分比较动员前明显降低, 而外周血中二的比例则显升高；动员前，后骨髓中CD34^＋CD62L^＋和CD34^＋CXCR4^ 细胞的变化并不明显，而外周血中前明显增加，后则显减少。输入CD44^＋，CD49d^ ，CD62L^ 及CXCR4^ 的CD34^＋细胞的量与移植后血中中性粒细胞和血小板恢复的时间未表明有显的相关，结论：G－CSF动员可下调骨髓CD34^＋细胞的CD44，CD49d，CD62L及CXCR4的表达，从而进入外周血循环，输注这些细胞的临床意义有待累积更多病例的分析。     

The dose of granulocyte-colony-stimulating factor after chemopriming treatment does not influence apheresis yield of progenitor cells: a retrospective study of 91 cases

BACKGROUND: The optimal dose of post-chemotherapy granulocyte-colony-stimulating factor (G-CSF) administration before peripheral blood progenitor cell (PBPC) collection has not been determined as yet, although 5 microg per kg per day has been recommended as the standard dose. This study retrospectively analyzed the effect of G-CSF dose on peripheral blood CD34+ cell collection from 91 patients with hematologic malignancies. STUDY DESIGN AND METHODS: Various doses of G-CSF were administered after several chemotherapeutic PBPC mobilization regimens. According to the dose of G-CSF administered, patients were assigned to two groups. Group 1 included 46 patients who received a low dose of G-CSF (median, 3.6 [range, 2.8-4.6] microg/kg/day). Group 2 included 45 patients who received a standard G-CSF dose of 6.0 (5.5-8. 1) microg per kg per day. Patients in the two groups were matched for age, diagnosis, previous therapy, and chemotherapeutic PBPC mobilization regimens. RESULTS: No difference was observed in the median number of CD34+ cells harvested from each group.The number of leukapheresis procedures necessary to obtain a minimum of 3 x 10(6) CD34+ cells per kg was the same in both groups, and the percentage of patients who failed to achieve adequate PBPC collections was similar in the two groups. CONCLUSION: The administration of low-dose G-CSF after chemotherapy appears equivalent to administration of the standard dose in achieving satisfactory PBPC collection.This approach could allow significant savings in medical cost. A randomized and prospective study is necessary, however, to assess the validity of these conclusions.     

Growth factor mobilization and modulation of progenitor cell adhesion to stromal cells: role of VLA-4

Cellular interactions between hematopoietic progenitor cells and bone marrow (BM) stromal cells are mediated by cell adhesion molecules (CAM). In agreement with previous studies, our flow cytometric analysis of isolated CD34+ cells showed that VLA-4 expression was significantly (p < 0.001) higher on steady-state BM than on CD34+ cells from growth factor-mobilized peripheral stem cell (PSC) products. To determine whether the expression of VLA-4 on progenitor cells plays a role in their adhesion to stromal cells, we examined the binding of isolated CD34+ progenitor cells from BM (n = 14) and PSC (n = 10) products to BM stromal cells in the presence or absence of a neutralizing antibody to VLA-4. In these studies, similar kinetics of BM and PSC CD34+ cell adhesion to BM stromal cells were observed. However, neutralizing antibody to VLA-4 significantly inhibited BM CD34+ but not PSC CD34+ cell adherence to stromal cells, suggesting a role for alternative CAM in cell binding. Further, in long-term co-cultures of BM CD34+ cells with BM stroma, we observed a significantly higher number of colony-forming units granulocyte-macrophage (CFU-GM) released into the media following treatment with neutralizing antibody to VLA-4 than in untreated control cultures. In contrast, no difference in the frequency of nonadherent CFU-GM between antibody-treated and control long-term co-cultures of PSC CD34+ cells with BM stromal cells was observed. This suggests that VLA-4 expression on mobilized PSC versus BM CD34+ cells has biologic relevance for at least 2 weeks based on the long-term BM culture results. In summary, these data suggest that the decreased expression of VLA-4 may have a role in the mobilization of progenitor cells, in part, by regulating their adherence to stromal cells, although additional mediators of adhesion are also involved.     

基质细胞衍生因子-1及其受体在G-CSF诱导的造血干/祖细胞动员中的作用

目的探讨基质细胞衍生因子-1（SDF-1）及其特异性受体CXCR4在G-CSF诱导的造血干／祖细胞（HSPC）动员中的作用。方法应用酶联免疫吸附实验（ELISA）、免疫组织化学、流式细胞术等方法检测健康供者稳态及G-CSF动员过程中骨髓、外周血SDF-1／CXCR4的变化，并应用SDF-1中和性抗体阻断BALB／c小鼠SDF-1信号通路，进一步验证SDF-1／CXCR4在动员中的作用。结果G-CSF动员前骨髓和外周血的SDF-1浓度分别为（7．23±0．66）μg/L和（5．43±0．35）μg／L，动员后分别为（5．88±1．03）μg／L和（5．42±0．52）μg/L。动员后骨髓SDF-1蛋白水平下降（P＜0．05），骨髓和外周血之间的SDF-1浓度梯度消失（P＞0．05）；稳态骨髓、动员后骨髓和动员后外周血的CD34^＋ CXCR4^＋细胞在CD34^＋细胞群中的比例分别为（40．98±21．56）％、（65．80±24．68）％和（27．54±26．03）％。动员后CXCR4在骨髓CD34^＋胞上表达增加（P＜0．05），而外周血CD34^＋细胞CXCR4表达降低（P＜0．05）。SDF-1中和性抗体可降低G-CSF动员的BALB／c小鼠外周血成熟白细胞和祖细胞集落数量（P＜0．05）。结论骨髓中SDF-1水平的降低以及CXCR4在HSPC上表达的下降促进了G-CSF介导的动员的发生。     

Peripheral blood hematopoietic stem cell mobilization and collection efficacy is not an independent prognostic factor for autologous stem cell transplantation

BACKGROUND: The successful mobilization and collection of hematopoietic stem cells are dependent on a number of clinical factors such as previous chemotherapy and disease stage. The aim of this retrospective study was to determine whether the effectiveness of mobilization and collection is an independent prognostic factor for autologous stem cell transplantation outcome. STUDY DESIGN AND METHODS: A total of 358 patients who received transplants from January 2003 to December 2004 (201 male and 157 female patients, ages from 2.7 to 77.3 years with median of 53 years of age) underwent autologous hematopoietic stem cell collection after mobilization with granulocyte-colony-stimulating factor (G-CSF) or G-CSF plus chemotherapy priming. This retrospective study included patients with diagnoses of acute myelogenous leukemia, non-Hodgkin's lymphoma, Hodgkin's disease, multiple myeloma, and solid tumors. All patients underwent stem cell collection until a target or a minimum CD34+ cell dose was reached. Correlations were performed between stem cell mobilization and/or collection efficacy and transplantation outcomes. RESULTS: In general, both larger reinfused CD34+ cell dose and shorter number of days for the stem cell count to reach the minimum of 2 x 10(6) per kg CD34+ cells do not foster quicker engraftment. Reinfused CD34+ cell dose of less than 12 x 10(6) and number of days stem cell collection to reach this minimum CD34+ cell dose did not independently affect the overall survival (OS) or disease-free survival (DFS). CONCLUSION: The effectiveness of hematopoietic stem cell mobilization and collection as defined as number of days to reach a CD34+ cell dose of 2 x 10(6) per kg should not be used independently to forecast posttransplantation prognosis, engraftment, DFS, and OS.     

Peripheral blood progenitor cell collection after epirubicin, paclitaxel, and cisplatin combination chemotherapy using EPO-based cytokine regimens: a randomized comparison of G-CSF and sequential GM-/G-CSF

BACKGROUND: The peripheral blood progenitor cell (PBPC) mobilization capacity of EPO in association with either G-CSF or sequential GM-CSF/G-CSF was compared in a randomized fashion after epirubicin, paclitaxel, and cisplatin (ETP) chemotherapy. STUDY DESIGN AND METHODS: Forty patients with stage IIIB, IIIC, or IV ovarian carcinoma were enrolled in this randomized comparison of mobilizing capacity and myelopoietic effects of G-CSF + EPO and GM-/G-CSF + EPO following the first ETP chemotherapy treatment. After ETP chemotherapy (Day 1), 20 patients received G-CSF 5 microg per kg per day from Day 2 to Day 13 and 20 patients received GM-CSF 5 microg per kg per day from Day 2 to Day 6 followed by G-CSF 5 microg per kg per day from Day 7 to Day 13. EPO (150 IU per kg) was given every other day from Day 2 to Day 13 to all patients in both arms of the study. Apheresis (two blood volumes) was performed during hematologic recovery. RESULTS: The magnitude of CD34+ cell mobilization and the abrogation of patients' myelosuppression were comparable in both study arms; however, GM-/G-CSF + EPO patients had significantly higher CD34+ yields because of a higher CD34+ cell collection efficiency (57.5% for GM-/G-CSF + EPO and 46.3% for G-CSF + EPO patients; p = 0.0009). Identical doses of PBPCs mobilized by GM-/G-CSF + EPO and G-CSF + EPO drove comparable hematopoietic recovery after reinfusion in patients treated with identical high-dose chemotherapy. CONCLUSION: The sequential administration of GM-CSF and G-CSF in combination with EPO is feasible and improves the PBPC collection efficiency after platinum-based intensive polychemotherapy, associating high PBPC mobilization to high collection efficiency during apheresis.