Autologous peripheral blood progenitor cell transplantation

Harvesting of autologous peripheral blood stem cells (PBSCs) has been facilitated by using currently available, efficient apheresis technology at the time of rebound from chemotherapy while patients are receiving recombinant growth factors, i.e., granulocyte (G) or granulocyte-macrophage (GM) colony stimulating factor (CSF). Ideally pheresis should be done before patients have had extensive stem cell toxins, i.e., alkylating agents or nitrosoureas. This strategy has facilitated the use of high dose chemoradiotherapy given as a single regimen or in a divided dose for patients with solid tumors or hematologic malignancies and results in more rapid engraftment than bone marrow transplantation (BMT). Although mere are no assays which measure repopulating stem cells, enumeration of CD34⁺ cells within PBSCs is a direct and rapid assay which provides an index of both early and late long-term reconstitutive capacity, since it correlates with colony-forming unit (CFU)-GMs, as well as pre-progenitor or delta assays and long-term culture-initiating cells (LTC-IC). A threshold of ≥2 × 10⁶ CD34⁺ cells/kg recipient body weight has been reported to be required for engraftment, but may vary depending upon the clinical setting. Strategies for mobilization of normal PBSCs also increase tumor cell contamination within PB in the setting of both hematologic malignancies and solid tumors, but the significance of these tumor cells in terms of patient outcome is unclear. Recently isolation of CD34⁺ cells from PBSCs has been done using magnetic beads or immunoabsorption on columns or rigid plates in order to enrich for normal hematopoietic progenitors and potentially decrease tumor cell contamination. As for other cellular blood components, standards have been developed to assure efficient collection and processing, thawing, and reinfusion, and to maintain optimal PBPC viability. Finally, future directions of clinical research include expansion of hematopoietic progenitor cells ex vivo; use of umbilical cord or placenta as rich sources of progenitor cells; syngeneic hematopoietic stem cell transplantation; related and unrelated allogeneic hematopoietic stem cell transplantation; treatment of infections, i.e., Epstein Barr virus, or tumor relapse after allogeneic BMT using donor PBSC infusions; and gene therapy approaches.     

Minimal number of circulating CD34+ cells to ensure successful leukapheresis and engraftment in autologous peripheral blood progenitor cell transplantation

BACKGROUND: The number of peripheral blood (PB) CD34+ cells has been widely used to monitor the timing of leukapheresis for autologous transplantation. However, no cutoff value for CD34+ cells in PB has been defined as a guideline for the identification of patients in whom the harvest would be effective and those in whom there was a high probability of failure. STUDY DESIGN AND METHODS: The present study investigated the best threshold of CD34+ cells in PB for successful harvesting and engraftment, using 263 PB samples with their corresponding leukapheresis components. In addition, that measure has been compared to other commonly used criteria such as the white cell count, the number of mononuclear cells, and the number of colony- forming units-granulocyte macrophage in PB. RESULTS : Time to engraftment of both granulocytes and platelets was significantly influenced by the number of CD34+ cells transfused, but all patients receiving > or = 0.75 × 10(6) CD34+ cells per kg achieved engraftment within a reasonable number of days (> 0.5 × 10(9)/L granulocytes by Day 11 and > 20 × 10(9)/L platelets by Day 13). A clear correlation between the number of CD34+ cells per microL in PB and of CD34+ cells per kg collected was found at each apheresis (r = 0.9, p < 0.0001). Moreover, the number of CD34+ cells per microL measured in PB the day the first leukapheresis was initiated displayed an excellent correlation with the total amount of CD34+ cells per kg finally collected (r = 0.81, p < 0.0001). On the basis of the regression curve obtained and the clinical engraftment results, it was found that the presence of > 5 CD34+ cells per microL in PB ensured a good yield from the harvest in 95 percent of patients and would avoid an unsuccessful harvest in 81 percent of cases. CONCLUSION: A dose of only 0.75 × 10(6) CD34+ cells per kg guarantees hematopoietic recovery within a reasonable number of days. To initiate a leukapheresis from which enough progenitor cells may confidently be obtained, a minimum of 5 CD34+ cells per microL in PB is required.     

Autologous graft-versus-host disease after CD34+-purified autologous peripheral blood progenitor cell transplantation

Autologous graft-versus-host disease (GVHD) has been frequently reported after cyclosporine A (CsA) administration in the autologous setting. This complication is related to the disruption of self-tolerance mechanisms induced by CsA and may exert an antitumor effect. We report the spontaneous occurrence of autologous GVHD after CD34+-purified peripheral blood progenitor cell transplantation (PBPCT) in 5 out of 24 consecutive patients (20.8%). The syndrome was characterized by skin rash (5/5), pruritus (5/5), eosinophilia (5/5), and fever (2/5) occurring at a median of 37 days (range 22-60) after transplantation. Diagnosis was confirmed by skin biopsy in all patients. The syndrome was self-limiting, lasted a median of 25 days, and did not require treatment. The rate of autologous GVHD was high after CD34+-purified autologous PBPCT. In fact, no autologous GVHD was documented in an historical control of 100 consecutive patients submitted to unmanipulated PBPCT at the same institution. The manipulation of the graft by the purging procedure causes a profound T lymphocyte depletion, thus possibly perturbing the equilibrium between autoregulatory cells and autocytotoxic T cells. These observations add new interest to the antitumor efficacy of autologous GVHD and suggest new questions regarding the role of transplantation for autoimmune diseases.     

Reticulated platelet monitoring after autologous peripheral haematopoietic progenitor cell transplantation

Reticulated platelets are the youngest marker of megakaryopoiesis. We monitored 13 haematological patients undergoing autologous peripheral haematopoietic progenitor cell transplantation. We used a flow cytometric method based on thiazole orange platelet staining. Patients showed a fall in reticulated platelet percentage at a mean of 5 days before a nadir of the platelet count. In all cases, platelet recovery is preceded by a rise of reticulated platelets; in 10 cases patients received platelet prophylactic transfusions after RP rise. The rise in reticulated platelets is an early sign of engraftment and may represent an easy parameter for transfusion management of transplanted patients.     

Changes in platelet demand in association with the spread of autologous peripheral blood progenitor cell transplantation.

Changes in platelet demand accompanying widespread application of autologous peripheral blood progenitor cell transplantation (APBPCT) were anticipated. The differences in the transfused volume of platelet concentrate (PC) among 8 patients with malignant lymphoma who were treated with APBPCT and 10 patients with malignant lymphoma who were not treated with APBPCT, although peripheral blood progenitor cell harvests had been performed, were studied. The former was 81 Japanese PC units more than the latter. Considering the supplied volume of PC from Red Cross blood centers and the number of APBPCTs in 1996 in Japan, the Japanese demand for PC increases by 0.16% for enforced APBPCT versus 0.36%, which includes the PC demand necessary in all treatment courses. The total increment in PC demand associated with APBPCT is not large enough to threaten the PC supply of Japan even if the number of APBPCTs increases rapidly.     

Successful autologous peripheral blood stem cell transplantation using thiotepa in a patient with systemic sclerosis and cardiac involvement

A 19-year-old man with systemic sclerosis (SSc) was hospitalized for autologous peripheral blood stem cell transplantation (auto-PBSCT) due to progressive scleroderma and cardiac involvement despite conventional treatment. During the administration of cyclophosphamide (60 mg/kg/day for 2 days) for mobilization and collection of CD34+ selected peripheral blood stem cells, he developed congestive heart failure. Echocardiogram showed hypokinetic asynergy from the septum to posterior wall, which might indicate underlying cardiac damage. We were also concerned about the risk of high-dose cyclophosphamide-induced cardiotoxicity. Since the dose-limiting toxicity of thiotepa, an alkylating agent, is myelosuppression, and cardiac toxicity due to thiotepa is less common, we used a conditioning regimen consisting of thiotepa (10 mg/kg/day, day -5) and low-dose cyclophosphamide (50 mg/kg/day, days -3 and -2), instead of the conventional high-dose cyclophosphamide (50 mg/kg/day x 4 days/course). The post-transplant course was uneventful, and the modified Rodnan skin thickness score improved from 32 to 15. The present case report demonstrates that thiotepa can be employed as a conditioning regimen for auto-PBSCT in SSc patients with cardiac involvement in order to reduce cyclophosphamide-induced cardiotoxicity.     

Automated red blood cell exchange in preparation for filgrastim mobilization of autologous peripheral blood hematopoietic progenitor cells in a patient with sickle cell anemia

Increasing survival of patients with sickle cell anemia (SCA) well into adulthood results in a rising likelihood of developing hematological malignancy. High‐dose chemotherapy with autologous hematopoietic progenitor cell (HPC) rescue is standard of care for several hematological malignancies, but the risk of severe or life‐threatening vaso‐occlusive phenomena during filgrastim mobilization of HPC for collection poses a potential barrier to this approach. We report the use of automated red cell exchange in preparation for filgrastim mobilization in a patient with homozygous SCA. Red cell exchange was repeated just prior to high‐dose chemotherapy to mitigate the need for red cell transfusion during bone marrow reconstitution. The patient experienced no vaso‐occlusive phenomena throughout the entire episode of care and did not become iron overloaded. This approach should be considered for all patients with homozygous or compound heterozygous sickle cell disease who are candidates for auto‐HPC rescue therapy.     

Leukapheresis after high-dose chemotherapy and autologous peripheral blood progenitor cell transplantation: a novel approach to harvest a second autograft

BACKGROUND: Autologous peripheral blood progenitor cells (PBPCs) are usually collected after the administration of conventional-dose chemotherapy (CDCT) and growth factors. However, there are no data available concerning the collection of PBPCs after high-dose chemotherapy (HDCT) and autologous hematopoietic transplantation in a larger series. STUDY DESIGN AND METHODS: Patients (n = 30) underwent leukapheresis for PBPC harvest after CDCT. After HDCT and autografting, the collection of a second PBPC autograft was attempted. RESULTS: Leukapheresis was performed after CDCT in all cases at a median of 118 CD34+ cells per microL (range, 18-589) and resulted in 6.4 x 10(6) CD34+ cells per kg (range, 1.7-29.0). After HDCT and autografting, 24 patients (80%) underwent secondary leukapheresis, although they had a significantly lower median of peripheral blood (PB) CD34+ cells (30/microL; range, 10-171; p < 0.001). In these patients a median of 3.6 x 10(6) CD34+ cells per kg (range, 1.6-10.1) was collected in the post-transplantation course. In the remaining six patients (20%) with PB CD34+ cells < 10 per microL, no PBPC harvesting was performed. These so-called poor mobilizers had received significantly less CD34+ cells for autologous transplantation than patients with successful post-HDCT mobilization (median, 2.5 x 10(6)/kg [range, 1.7-3.0] vs. 6.5 x 10(6)/kg [range, 3.2-19.6]; p < 0.001). CONCLUSION: Collection of PBPCs is possible in most patients during the recovery phase of hematopoiesis after HDCT plus autografting, and the number of circulating PBPCs may be related to the CD34+ cell dose transfused by the preceding autograft.     

Large-volume apheresis for the harvest of peripheral blood progenitor cells for autologous transplantation

BACKGROUND: The mobilization and harvest of a sufficient number of peripheral blood stem and progenitor cells for autologous transplantation is an important aspect of treatment in patients with certain hematologic and solid tumor disease. The level of CD34+ cells in peripheral blood is often used as a predictor of successful harvest. STUDY DESIGN AND METHODS: A total of 129 apheresis procedures in 38 patients have been investigated retrospectively to evaluate the possibility to predict the outcome by other measures, such as total treated blood volume (TBV) during the apheresis. RESULTS: No significant correlation was observed between the level of CD34+ cells per kg of body weight in collected apheresis components and the TBV in all 129 apheresis procedures. However, analysis of results from 22 apheresis procedures with TBV > 16 L (large-volume apheresis) and with < 10 × 10(3) CD34+ cells per mL in the peripheral blood found a correlation between TBV and the number of CD34+ cells per kg of body weight in the collected component (R2 = 0.585, p = 0.005). In patients who underwent large-volume apheresis (> 16 L) and who had < 10 × 10(3) CD34+ cells per mL in their peripheral blood, the number of CD34+ cells in the apheresis component was not correlated with that in the peripheral blood prior to harvest (R2 = 0.262, p = 0.1569). In the patients who underwent apheresis procedures with TBV < 16 L and who had > 20 × 10(3) CD34+ cells per mL in their peripheral blood, there was a correlation between the number of CD34+ cells in the component and the number of CD34+ cells in the peripheral blood (R2 = 0.800, p = 0.0000). However, there was not a correlation in this group between the number of CD34+ cells in the component and the TBV. There were no significant differences in the content of CD34+/CD33+ and CD34+/ HLA-DR+ cells in the collected component in the two groups. CONCLUSION: TBV appears to be critical for the collection of a sufficient number of progenitor cells in patients with < 10 × 10(3) CD34+ cells per mL in peripheral blood.     

自体外周血造血干细胞移植治疗恶性血液病的临床研究

目的：探讨自体外周血造血干细胞移植（ＡＢＳＣＴ）治疗恶性血液病的疗效。方法：ＡＢＳＣＴ１３例，自体外周血干细胞（ＰＢＳＣ）及自体骨髓联合移植１５例。移植前采用化疗加多抗甲素或粒细胞集落刺激因子（ＧＣＳＦ）动员外周血干细胞。结果：多抗甲素组每例采集单个核细胞数为（３．５８±２．１２）×１０８／ｋｇ，ＧＣＳＦ组为（６．６８±５．３１）×１０８／ｋｇ；接受移植的２８例患者中２１例呈持续缓解状态（ＣＣＲ），中位ＣＣＲ时间为１８（５～５８）个月，复发７例；３年无病生存率为６８．７％，复发率为２２．３％。结论：两种动员方案均有良好的动员效果；ＡＢＳＣＴ及ＰＢＳＣ和自体骨髓联合移植均具有良好疗效及造血功能恢复快、合并症少等优点，值得进一步推广应用。     

自体外周血造血干细胞移植患者的护理

目的探讨对行自体外周血造血干细胞移植的患者在不同阶段实施的相应护理措施。方法选取我院2005年10月至2008年12月进行APBSCT治疗的恶性肿瘤患者60例,根据其移植的各个阶段采取相应的护理措施。结果通过施行全环境保护、合理饮食、全身皮肤黏膜清洁消毒、心理支持等护理活动,有效地减少了移植过程中各种并发症的发生率。结论在进行APBSCT治疗的患者中,护理质量的优劣是直接影响APBSCT成败和患者预后好坏的重要因素,良好的护理使癌症患者的生命得到真正的延长。     

儿童自体外周血造血干细胞移植的护理

自体外周血造血干细胞移植(APBSCT)是对患者进行超大剂量化疗或全身放疗预处理后,将正常外周血造血干细胞植入患者体内,使其重建正常造血及免疫功能的治疗方法[1].由于儿童对大剂量化疗和全身照射治疗不良反应敏感性较高,对护理工作提出了更高要求.     

非血缘异基因外周血造血干细胞移植治疗急性粒细胞白血病一例

目的：开展非血缘关系供者外周血造血干细胞移植（ＰＢＳＣＴ）对血液恶性肿瘤进行根治性治疗，并观察其长期造血重建和移植物抗宿主病（ＧＶＨＤ）及一些移植并发症。方法：选择ＨＬＡ相配非血缘关系的２６岁男性健康供者的外周血干细胞移植给１例急性粒细胞白血病第１次复发缓解后的１１岁男性患儿。预处理方案为分次全身６０Ｃｏ照射（８Ｇｙ）加环磷酰胺（１２０ｍｇ／ｋｇ）和足叶乙甙（３０ｍｇ／ｋｇ）。移植物抗宿主病预防用环孢霉素Ａ（ＣｓＡ）联合氨甲喋呤（ＭＴＸ）。移植单个核细胞为８．３×１０８／ｋｇ，ＣＤ３４＋细胞为１．０７９×１０８／ｋｇ，ＣＦＵ－ＧＭ为１．０９×１０６／ｋｇ。结果：＋１０天中性粒细胞达１．３０×１０９／Ｌ，＋１８天骨髓显示造血重建，＋２３天ＤＮＡＤ１Ｓ８０显示移植物植入成功，＋１２０天受者血型由ＡＢ型转为供者血型Ａ型。＋１１天出现Ⅰ度急性ＧＶＨＤ，＋１４３天出现慢性ＧＶＨＤ，分别用甲基泼尼龙和泼尼松控制。随访３００余天，患者情况正常。结论：非血缘ＰＢＳＣＴ可以维持长期造血     

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.     

Hematopoietic progenitor cell count, but not immature platelet fraction value, predicts successful harvest of autologous peripheral blood stem cells

Mobilized stem cells in the peripheral blood (PB) must be efficiently harvested at the appropriate time before autologous PB stem cell (PBSC) transplantation. Enumeration of CD34+ cells in the PB before apheresis predicts the number of PBSCs that can be collected, but the cytometric techniques used are complex and expensive. Therefore, it is necessary to identify an alternative to the CD34+ cell count in PBSC harvest-time monitoring. Fully automated flow cytometry using blood cell counters now allows reliable quantification of immature myeloid cells in the PB, referred to as hematopoietic progenitor cells (HPC), and reticulated platelets, expressed as the immature platelet fraction (IPF). Immature or reticulated platelets are thought to correlate with thrombopoietic activity of the marrow. Following a chemotherapy nadir, the recovery of white blood cell and platelet counts has been used to determine the right time for apheresis. Therefore, we examined whether the HPC count and IPF value could be used to predict PBSC mobilization in 20 patients with hematological malignancies. The HPC count was found to be correlated with the CD34+ cell count (r = 0.84, P < 0.01), whereas the IPF value was not (r = 0.37, P = 0.44). Therefore, the HPC count, but not the IPF value, is a possible predictor of the timing of autologous stem cell transplantation.     

自体外周血干细胞移植病人的健康教育

[目的]为56例进行自体外周血造血干细胞移植(APBSCT)的病人提供全程的健康教育,使病人在层流病房中很好地配合治疗及护理,减少并发症,顺利转出层流病房.[方法]采取一对一交谈法、病人现身说法等形式,从病人心理、层流室环境、无菌观念、化疗、回输造血干细胞过程、Ⅳ度骨髓抑制期、转出层流室和出院进行全面的健康教育.[结果]56例病人在干细胞移植的各个阶段都能积极配合医护工作,平均15.2 d就完成整个自体外周血造血干细胞移植过程,顺利转出层流病房.[结论]56例病人经过全程健康教育,能正确认识自体外周血干细胞移植的过程及在层流室中配合医护工作,心身协调地完成治疗.