Treatment and follow‐up of 18 severe systemic lupus erythematosus patients with stem cell transplantation

Aim: To investigate the feasibility, efficacy and safety of high‐dose immunosuppressive therapy and autologous peripheral blood stem cell transplantation (PBSCT) with CD34⁺ cell selection in patients with refractory and severe autoimmune diseases. Methods: Eighteen patients with persistent systemic lupus erythematosus refractory to conventional treatment were enrolled into the study of peripheral blood stem cell transplantation in Peking Union Medical College Hospital from 1999 to 2005. After mobilization and conditioning, the enriched CD34⁺ cells were reinfused. Disease activity, adverse effects, haematopoietic and immunologic reconstitution were monitored and followed up for at least 6 months. Results: Overall treatment‐related mortality was 5.6% with one patient dying of cytomegalovirus infection. The overall remission rate was 95.8% in the first year after PBSCT. Relapse occurred in three patients (17.6%) in 37, 26, and 19 months post‐transplantation, respectively. Disease Activity Index scores of systemic lupus erythematosus survivors were decreased significantly (P  <  0.001). Conclusions: Short‐term effect of autologous peripheral blood stem cell transplantation is promising although treatment‐related mortality and relapses are observed in a subset of patients. High‐dose immunosuppressive therapy followed by autologous peripheral blood stem cell transplantation with CD34⁺ cell selection is feasible and relatively safe in the treatment of severe and refractory autoimmune diseases. The long‐term effect needs further evaluation and multicentre study.     

Comparison of myeloma cell contamination of bone marrow and peripheral blood stem cell harvests

It could be speculated for patients with myeloma and other lymphoproliferative disorders that peripheral blood stem cells may be preferable to bone marrow for autologous transplantation because they may be less contaminated by neoplastic cells. To test this possibility, the immunoglobulin heavy chain gene rearrangement and limiting dilution polymerase chain reaction were used to sensitively quantify myeloma cells in bone marrow and peripheral blood stem cell collections, taken at a similar time, from eight patients with multiple myeloma. Levels of residual disease in the peripheral blood stem cell harvests were variable and did not reflect the tumour burden in the marrow. Peripheral blood stem cells contained 1.7 to 23 700-fold fewer myeloma cells compared with the bone marrow and would have resulted in reinfusion of 0.08 to 59 480-fold fewer myeloma cells based on total reinfused CFU-GM and 0.24 to 24 700-fold fewer myeloma cells based on total reinfused nucleated cells. Assuming that the proportion of clonogenic myeloma cells is equivalent, peripheral blood stem cells may be better than bone marrow as a source of haemopoietic stem cells for transplantation in multiple myeloma. The clinical follow-up suggested that patients transplanted with peripheral blood stem cells containing a low number of myeloma cells had better disease control than those transplanted with peripheral blood stem cells containing a high number.     

Factors which affect the CFU-GM content of the peripheral blood haemopoietic progenitor cell harvests in patients with acute myeloid leukaemia

Autologous peripheral blood haemopoietic stem cells (PBSC) were harvested from 30 patients with de novo acute leukaemia, 29 of whom had entered remission following standard chemotherapy. Correlation of CD34⁺ cells/kg to CFU-GM/kg in the harvests was good (correlation coefficient=0.72, P<0.001). We demonstrated significant associations between the CFU-GM content of the harvest and the following: time to platelets >50×10⁹/l post final induction course (P<0.0001), days to harvest from day 1 of intensification/mobilization (correlation coefficient=?0.73, P<0.001), platelets >20×10⁹/l at time of harvest (P=0.02), time to WBC >1.0×10⁹/l post intensification/mobilization (correlation coefficient=?0.70, P<0.001), and WBC on day of harvest (correlation coefficient=0.60, P<0.001). In contrast, we found no relationship between the CFU-GM content of the harvest and patient age up to 65 years, presence of absence of coexistent features of trilineage myelodysplasia at diagnosis, number of induction courses to remission or total number of courses of chemotherapy prior to intensification/mobilization. Haemopoietic recovery after reinfusion of PBSC was highly correlated to the number of CFU-GM infused (neutrophils >0.5×10⁹/l r_s=?0.72, P=0.001; platelets >20×10⁹/l unsupported r_s=?0.71, P=0.001). Our results show that the number of induction courses received, and thus exposure to cytotoxic agents received, made no significant difference to subsequent CFU-GM harvest content. We collected superior harvests from those patients with faster platelet recovery following mobilization therapy. We also found that faster platelet recovery following the final induction therapy was a better predictor of the CFU-GM harvest following mobilization than was the neutrophil recovery following final induction.     

G-CSF动员同胞供体外周血干细胞的研究

目的：研究粒细胞集落刺激因子（Ｇ－ＣＳＦ）动员异基因外周血干细胞（ＰＢＳＣ）的动力学,分析移植物细胞成分,探讨供者年龄、特别对动员效果的影响。方法：４０个ＨＬＡ相合的同胞供者一日两次皮下注射Ｇ－ＣＳＦ５μｇ／ｋｇ,于第５、６天采集ＰＢＳＣ,所得ＰＢＳＣ以流式细胞仪分析免疫表型。结果：①Ｇ－ＧＳＦ动员过程中供者耐受良好,外周血白细胞和单个核细胞（ＭＮＣ）峰值在第５天;②ＰＢＳＣ含有ＭＮＣ５．８（３．     

造血干细胞移植供者应用粒细胞集落刺激因子的研究

目的　探讨外周血造血干细胞移植 (PBSCT)供者应用粒细胞集落刺激因子 (G CSF)后细胞成分的变化和药物对供者身体状况的近期影响。方法　对 18例健康PBSCT供者给予G CSF 5～ 10 μg/ (kg·d) ,4～ 5d ,采集外周血单个核细胞进行检测 ,临床观察用药后出现的毒副反应。结果　①外周血白细胞在动员后 4～ 5d达峰值 ,动员后比动员前高 7～ 14倍 (P <0 0 1)。②细胞采集在动员后第 4～ 5天开始 ,两者单个核细胞 (MNC)和CD34 细胞值之间无明显差异。③本组供者不同性别和体重间 ,MNC值有显著差异性。④主要毒副反应为骨痛和肌痛、乏力等 ,采集细胞过程中出现口唇、四肢麻木。结论　对PBSCT供者应用G CSF 5～ 10 μg/ (kg·d) ,4～ 5d可有效动员MNC和CD34 细胞。供者对此剂量的毒副反应能够耐受     

类风湿关节炎患者外周血CD34+造血干/祖细胞的检测及其意义

目的 探讨类风湿关节炎(RA)外周血造血干/祖细胞(HSC/HPC)的数量及细胞膜CD34平均荧光强度(MFI)变化及其与临床指标的关系,以期阐明其在RA中的意义.方法 收集34例RA患者和16名健康对照者外周血,利用单克隆抗体标记CD34+细胞,流式方法测定CD34+HSC/HPC所占外周血淋巴细胞的比例及膜CD34的MFI,分析其与外周血细胞计数、疾病活动病程和药物应用的关系.数据分析采用t检验和方差分析,相关性分析采用Pearson相关分析.结果 ①RA患者CD34+细胞在外周血中淋巴细胞中所占的比例比健康人偏低[分别为(0.13±0.09)%和(0.38±0.21)%,P＜0.05],但两者所占外周血淋巴细胞的比例均低于0.5%;但MFI偏高(分别为57±33和31±11,P＜0.05).②RA患者CD34+细胞在外周血淋巴细胞中所占的比例与外周血红细胞计数、血红蛋白浓度呈正相关性,和C反应蛋白呈负相关;其MFI与健康评价问卷表(HAQ)评分、X线分期呈正相关,与血小板计数呈负相关.③RA患者CD34+细胞在外周血淋巴细胞中所占比例的降低程度以及MFI与疾病的活动性、病程和用药情况无明显相关性(P＞0.05).结论 造血干细胞可能在RA的发病机制中起作用.     

The endogenous granulocyte colony-stimulating factor response following autologous peripheral blood stem cell transplantation is impaired in patients with myeloma

Granulocyte colony-stimulating factor (G-CSF) levels were studied in 23 patients (10 myeloma, 13 relapsed Hodgkin's disease, non-Hodgkin's lymphoma or germ cell tumours), post autologous peripheral blood stem cell transplantation (PBSCT). The two groups had similar previous chemotherapy and numbers of CD34+ cells transplanted. All patients received G-CSF by injection starting 8 d post transplantation. Twenty out of 23 patients showed raised endogenous levels of G-CSF before cytokine administration. Myeloma patients showed significantly lower levels of endogenous G-CSF than the other patients (0.767 versus 3.262 ng/ml, P < 0.05). Further rises in G-CSF levels were seen following the administration of exogenous G-CSF which then fell, despite ongoing administration of G-CSF, as neutrophil recovery occurred.     

Successful treatment with allogeneic peripheral blood stem cell transplantation and granulocyte transfusion for severe aplastic anemia with sinusitis

A 43-year-old woman with severe aplastic anemia (SAA) received anti-thymocyte globulin and cyclosporin A (CyA) and achieved hematological remission. Although she had maintained hematological remission, the disease relapsed 10 months after arbitrary discontinuance of maintenance therapy with CyA. Resumption of CyA therapy was not effective, and her condition became complicated with progressive sinusitis with bone destruction, which was refractory to antibiotics, antifungal agents, granulocyte colony-stimulating factor, and surgical drainage. Because of the necessity for early neutrophil recovery (to resolve the infection), we proceeded with a combination therapy using allogeneic peripheral blood stem cell transplantation (PBSCT) promptly followed by granulocyte transfusion (GTX) from the same human leukocyte antigen-identical donor rather than carrying out a second immunosuppressive therapy. The patient showed temporal resolution of infection on the second day after a single GTX. Although the patient had pneumonia on day 11, it was resolved promptly after engraftment on day 16. This report suggests the clinical utility of a salvage therapy with allogeneic PBSCT followed by GTX in a particular case of recurrent SAA with refractory infections.     

Autologous peripheral blood stem cell transplantation (PBSCT) in acute myeloid leukaemia (AML) in first complete remission (CR) a report of the BGMT cooperative group about 32 patients

Thirty-two patients < 55 year with AML in first complete remission underwent autologous peripheral blood stem cell transplantation (PBSCT). Before transplant, they were treated with one or two induction courses plus one consolidation cycle including daunorubicin and araC. Thereafter, haemopoietic stem cells (HSC) were mobilized using daunorubicin 45 mg/m² days 1-3 plus either high-dose araC 3 g/m² TID, 4 days (BGMT87 trial, 13 patients) or intermediate-dose araC 500 mg/m² TID, 4 days and G-CSF from day 7 until completion of cytaphereses (BGMT91 trial, 19 patients). The median number of leukaphereses was 6 (range 5-6). Collections were better in the BGMT91 protocol concerning median MNC (18.1 versus 9.1 10⁸/kg, p 0.0019), median CFU-GM (36.5 versus 13.5 10⁴/kg, p 0.017), and median CD34+ cells (12.2 versus 7.65 10⁶/kg, p 0.24). The unpurged product was reinfused after busulfan 16 mg/kg and melphalan 140 mg/m². All but one patients engrafted. Number of days with fever and antibiotics, and hospitalization duration were significantly reduced in the BGMT91 protocol. No death in aplasia was encountered. The actuarial DFS at 3 years was 55%+/-18%. Sufficient HSC can be collected in AML following intensive consolidation and autologous PBSCT represents a valuable procedure in this setting.     

非清髓性自体外周血造血干细胞移植治疗系统性红斑狼疮远期疗效分析

目的探讨非清髓性自体外周血造血干细胞移植(NAST)治疗系统性红斑狼疮(SLE)的远期疗效。方法总结中山大学附属第五医院2002年11月至2005年10月4例成功接受NAST的SLE患者移植后的随访情况。非清髓性预处理移植前1～2 d静脉滴注阿糖胞苷200 mg/(kg.d)及环磷酰胺40 mg/(kg.d)。评价患者移植前后的相关症状体征、远期并发症及免疫功能的变化。结果白细胞总数恢复正常的中位时间12 d,血小板>100×109/L的中位时间为10 d,血红蛋白>120 g/L的中位时间为22 d。随访中,NAST后4例患者临床症状和体症均消失,淋巴细胞亚群检测显示:CD4+及CD4+/CD8+均恢复正常。1例男性患者移植4年后妻子正常受孕并产下一健康女婴。3例女性均恢复正常工作与生活。结论 NAST造血重建快,远期疗效确切。SLE患者NAST治疗后生活质量较好。     

Factors associated with granulocyte colony-stimulating factor-induced peripheral blood stem cell yield in healthy donors

BACKGROUND AND OBJECTIVES: Poor collection results are a clinical problem in granulocyte-colony stimulating factor (G-CSF)-induced peripheral blood stem cell (PBSC) collection in healthy donors. It would be beneficial to be able to predict the PBSC yield from allogeneic donors before mobilization or harvesting. MATERIALS AND METHODS: We examined the relationship between certain donor characteristics and the effectiveness of G-CSF-induced PBSC collection in 59 healthy family donors aged 3-63 years old (median 16 years). G-CSF was administered subcutaneously at 10 microg/kg for mobilization, daily for 5 days, and PBSC harvest using a continuous blood cell separator was started on day 5 of G-CSF treatment. Total cell yields were calculated as the number per unit of processed blood (l) per unit weight of the donor (kg). RESULTS: In a univariate analysis, the donor's age, body mass index (BMI), white blood cell (WBC) count before mobilization, and platelet count before and during mobilization were significantly correlated with the yield of mononuclear cells (MNC), CD34(+) cells and granulocyte-macrophage colony-forming units (GM-CFU). Younger age (P < 0.001), a low BMI (P = 0.002), a high WBC count before mobilization (P = 0.004), a high platelet count before (P = 0.012) and during (P < 0.05) mobilization, and a low speed of withdrawal (P = 0.019) were associated with a higher CD34(+) cell yield. No significant correlation was found for gender, the type of G-CSF, the serum level of G-CSF, the type of cell separator, or the type of blood access. A multivariate forward and backward stepwise selection regression analysis showed that the factors associated with CD34(+) cell yield were age, platelet count before and during mobilization, and circulating CD34(+) cell concentration on day 2 of G-CSF treatment. CONCLUSION: In this small preliminary study, we found that donor age is the most important factor in predicting G-CSF-induced PBSC yields. Old age and low platelet counts before mobilization might be useful indicators for identifying poor mobilizers. Further validation of these findings in a larger number of donors are needed to establish whether these findings apply to other populations.     

Successful collection of peripheral blood progenitor cells in patients with acute myeloid leukaemia following early consolidation therapy with granulocyte colony-stimulating factor-supported high-dose cytarabine and mitoxantrone

We evaluated the feasibility of collecting peripheral blood progenitor cells (PBPC) in patients with acute myeloid leukaemia (AML) following two cycles of induction chemotherapy with idarubicin, cytarabine and etoposide (ICE), and one cycle of consolidation therapy with high-dose cytarabine and mitoxantrone (HAM). Thirty-six patients of the multicentre treatment trial AML HD93 were enrolled in this study, and a sufficient number of PBPC was harvested in 30 (83%). Individual peak concentrations of CD34⁺ cells in the blood varied (range 13.1–291.5/μl; median 20.0/μl). To reach the target quantity of 2.5 × 10⁶ CD34⁺ cells/kg, between one and six (median two) leukaphereses (LP) were performed. The LP products contained between 0.2 × 10⁶ and 18.9 × 10⁶ CD34⁺cells/kg (median 1.2 × 10⁶/kg). Multivariate analysis showed that the white blood cell count prior to HAM and the time interval from the start of HAM therapy to reach an unsupported platelet count > 20 × 10⁹/l were predictive for the peak value of CD34⁺ cells in the blood during the G-CSF stimulated haematological recovery. In 16 patients an intraindividual comparison was made between bone marrow (BM) and PBPC grafts. Compared to BM grafts, PBPC grafts contained 14-fold more MNC, 5-fold more CD34⁺ cells and 36-fold more CFU-GM. A CD34⁺ subset analysis showed that blood-derived CD34⁺ cells had a more immature phenotype as indicated by a lower mean fluorescence intensity for HLA-DR and CD38. In addition, the proportion of CD34⁺/Thy-1⁺ cells tended to be greater in the PBPC grafts. The data indicate that sufficient PBPC can be collected in the majority of patients with AML following intensive double induction and first consolidation therapy with high-dose cytarabine and mitoxantrone.     

Plerixafor and Filgrastim XM02 (Tevagastrim) as a first line peripheral blood stem cell mobilisation strategy in patients with multiple myeloma and lymphoma candidated to autologous bone marrow transplantation

Plerixafor, a CXCR4 antagonist, has shown to be effective in increasing the number of circulating stem cells, even in patients failing a previous mobilisation attempt. Recently a number of non-glycosylated recombinant human granulocyte-colony stimulating factor (G-CSF) has been clinically approved for the same indications as the originator G-CSF for comparable safety and efficacy and their reduced cost. In an attempt to provide a less toxic strategy, 14 patients affected by haematological malignancies (non-Hodgkin's lymphoma = 4, Hodgkin's disease = 2 and multiple myeloma = 8), received the combination of biosimilar filgrastim and plerixafor as a first line mobilising strategy. The median number of circulating CD34+ cells on day 4 was 16 (3-42); Plerixafor was administered to all, but one patient who had already 42 CD34+ cells per μL on day 4. On day 5, after plerixafor administration, the median number of circulating CD34+ cells had raised to 60 per μL (14-138). All the patients underwent leukapheresis and were able to collect a number of CD34+ cells ≥ 2.0 × 10(6) /kg in a median number of procedures of one. Although preliminary, these data show the combination of biosimilar filgrastim and plerixafor is effective and provides a non-toxic approach to mobilise stem cells.     

De novo CD5+ diffuse large B cell lymphoma with basophilia in the peripheral blood: successful treatment with autologous peripheral blood stem cell transplantation

Basophils play an important role in allergic inflammation and are pathologically related to hematological disturbances, such as iron deficiency anemia and myeloproliferative disorders; however, they are only rarely encountered in lymphoid malignancies. Here, we report the case of a 33-year-old man with a bulky mass of the small intestine, multiple paraaortic lymphoadenopathy, pleural effusion, and ascites, who was diagnosed as a case of de novo CD5+ diffuse large B cell lymphoma (DLBCL). This patient showed a marked elevation of the basophil count in the peripheral blood, which appeared to run in parallel with the tumor burden. High dose chemotherapy followed by autologous peripheral blood cell transplantation yielded complete remission, and the patient has remained disease free for 5 years. To the best of our knowledge, this is the first report of a case of de novo CD5+ DLBCL showing marked elevation of the PB basophil count.     

Detection of monoclonal plasma cells in the peripheral blood stem cell harvests of patients with multiple myeloma

We evaluated the harvest product from 47 patients undergoing peripheral blood (BP) stem cell collections for monoclonal plasma cells (PC) using a sensitive immunofluorescence technique. 60% (28/47) had docu mented tumour cells in the apheresis product. The 32 patients in plateau had a mean of 1.62 x 10⁶ PC/1 v 74.64 x 10⁶ in 15 relapsed patients ( P < 0.01). 32% (6/19) of patients without any tumour cells in the initial sample had them detected on a subsequent apheresis sample. In four cases (all treated with GM-CSF) small numbers of tumour cells were detected initially but became undetectable on a subsequent sample.     

Fatal upper and lower gastrointestinal cytomegalovirus disease following autologous peripheral blood stem cell transplantation

Although the life-threatening cytomegalovirus (CMV) disease is a well known complication following allogeneic hematopoietic stem cell transplantation (HSCT), it has been considered infrequent after autologous peripheral blood stem cell transplantation (PBSCT). On the other hand, the massive involvement of the gastrointestinal (GI) tract as the primary site of fatal CMV disease is particularly rare after autologous PBSCT. We present the case of a woman who suffered from CMV disease after high-dose busulphan/melphalan/thiotepa (BuMelTT) and autologous PBSCT. The primary site of infection was the GI tract, which was extensively affected. During the fifth week post-transplant the patient started with epigastralgia, diarrhea, fever, GI bleeding, and thrombocytopenia, and she died on day +52. Another case of fatal CMV disease among the few patients treated with BuMelTT has been recently reported, which suggests that the immunodeficiency associated with that regimen can be as intense as that occurring after allogeneic BMT.     

Individually determined dosing of filgrastim after autologous peripheral stem cell transplantation in patients with malignant lymphoma--results of a prospective multicentre controlled trial

OBJECTIVES: To explore the safety and effectiveness of the individually determined application granulocyte-colony stimulating factor (G-CSF) after autologous peripheral blood stem cell transplantation (ASCT). METHODS: The administration of G-CSF from day +5 (arm A) was compared in a randomised, controlled trial with delayed, individually determined administration (G-CSF started when WBC >or= 0.5 x 10(9)/L and ANC >or= 0.1 x 10(9)/L or at day +10; arm B), and with placebo (arm C). RESULTS: One hundred and six patients, median age 45 (range 21-64), all with malignant lymphoma treated with BEAM chemotherapy were analysed. A significant difference in the time to neutrophil engraftment and in the duration of neutropenia <0.5 x 10(9)/L and <1.0 x 10(9)/L was observed between the arms (P = 0.04-<0.0001) with a 1-d prolongation of the median durations in arm B in comparison with arm A but a 2-4-d prolongation in the placebo arm C in comparison with arm B. The median number and range of days to neutrophil engraftment >0.5 x 10(9)/L after graft re-infusion was 10 (9-14) in arm A; 11 (9-19) in arm B; and 14 (10-30) in arm C (P < 0.0001). Engraftment of platelets to >20 x 10(9)/L and >50 x 10(9)/L was significantly delayed in the arms using G-CSF in comparison with placebo (P = 0.04-0.002) without any increase in bleeding or in transfusion requirement. There was no difference in the incidence and duration of transplant-related complications and their treatment between the arms. CONCLUSIONS: Our study has confirmed the safety of individually determined administration of G-CSF. The optimal timing of G-CSF application after ASCT in patients with good-quality grafts is shortly before expected spontaneous engraftment.     

Elevation of serum hepatocyte growth factor during granulocyte colony-stimulating factor-induced peripheral blood stem cell mobilization

We examined serum levels of the angiogenic factors, vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and hepatocyte growth factor (HGF), in normal donors for allogeneic peripheral blood stem cell (PBSC) transplantation. Granulocyte colony-stimulating factor (G-CSF) (filgrastim 400 microg/m2/d) was administered to 23 donors for 5 d and aphereses were performed on days 4 and 5. Although bFGF remained at similar levels after G-CSF treatment, serum VEGF and HGF levels increased 1.5-fold (n = 13; P = 0.02) and 6.8-fold (n = 23; P < 0.0001) respectively. The serum HGF level before G-CSF administration on day 1 correlated inversely with mobilized CD34+ cell numbers. Time course kinetics of HGF showed that on the day after G-CSF administration (day 2), serum HGF levels increased to 3678 pg/ml. For auto PBSC mobilization with chemotherapy and G-CSF 200 microg/m2/d (n = 8), we observed similar HGF elevation, which appeared to be dose-dependent on the G-CSF administered.     

Impact of different dimethyl sulphoxide concentrations on cell recovery,viability and clonogenic potential of cryopreserved peripheral blood hematopoietic stem and progenitor cells

Background and Objectives The procedure of autologous hematopoietic stem/progenitor cell transplantation requires cryopreservation. Addition of DMSO is necessary to secure the viability of such cells, but this solvent is potentially toxic to stem cells’ recipient. 10% DMSO solution is used by the majority of transplant centres. The aim of our study was to test if DMSO concentration might be reduced without negative impact on cell recovery and clonogenicity. Materials and Methods Samples were prospectively collected from 20 patients. Small volumes of leukapheresis products were frozen with different cryoprotective mixtures, containing 10%, 7·5%, 5% and 2·5% DMSO, respectively. The quality of cryoprotective mixtures was evaluated based on recovery, viability and clonogenic potential of hematopoietic stem cells after defreezing. Results Reduction in DMSO concentration to 7·5% or lower was associated with decreased recovery of nucleated cells. In contrast, the number of colonies was highest for 7·5% DMSO with significant differences when compared to 10% DMSO solution. Conclusion Reduction in DMSO concentration from 10% to 7·5% may have favourable impact on hematopoietic recovery after autologous transplantation. The findings require confirmation in a clinical setting.