Collection of peripheral blood stem cells from normal donors 60 years of age or older

We report 14 normal peripheral blood stem cell (PBSC) donors ≥ 60 years of age who had cytokine mobilization followed by PBSC apheresis for allogeneic transplantation. Mobilization was achieved with filgrastim (6 μg/kg twice daily). Their median age was 63.5 years (range 60–77), and 43% had a positive medical history, mainly hypertension and/or cardiac problems. Their median pre-apheresis leucocyte count (×10⁹/l) was 38.6 (range 29.6–63.4). The median apheresis yield (×10⁶ CD34⁺ cells/litre blood processed, first apheresis) was 27.9 (range 1.6–54.8). The target cell dose (≥4& times; 10⁶ CD34⁺ cells/kg recipient) was reached with one procedure in eight (57%) donors. Filgrastim-related adverse events were acceptable and apheresis was well tolerated. When compared to younger donors (<60 years of age), a trend to a lower CD34⁺ apheresis yield and a requirement for more than one apheresis to achieve the collection target (≥4 ×10⁶ CD34⁺ cells/kg) was evident. Although older (≥60 years) donors seem to mobilize less effectively, these data suggest that PBSC collection from them is feasible and has an acceptable short-term safety profile.     

异基因外周血造血干细胞移植治疗难治性白血病(附1例报告)

对 1例难治性急性粒 -单细胞白血病 (AML - M4 b)患者施行异基因外周血造血干细胞移植 (allo-PBSCT ) ,以 Cy/ TBI方案预处理后 ,输注 HL A完全相合的同胞供者经 G- CSF动员的外周血单个核细胞(PBMNCs) 9.0× 10 8/ kg,其中 CD34 细胞 6 .2 5× 10 6 / kg;移植物抗宿主病 (GVHD)的预防用 Cs A MTX方案。结果 : 15天时 ,外周血中性粒细胞 >0 .5× 10 9/ L,血小板 >5 0× 10 9/ L; 30天时 ,外周血三系均完全恢复正常。仅有 度皮肤 GVHD发生。认为对于难治性白血病 ,如有 HL A相合供者 ,应及早行异基因造血干细胞移植 (allo-HSCT)特别是 allo- PBSCT,具有受者造血与免疫功能重建快等优点     

Autologous transplantation in chronic myeloid leukaemia using peripheral blood stem cells

Forty-three patients with chronic myeloid leukaemia in first chronic phase were recruited to study intensive chemotherapy (idarubicin plus cytarabine; IdAC) followed by collection of peripheral blood stem cells (PBSC) in the recovery phase. PBSC autografting was performed on 32 patients. One patient died during mobilization and three died following autograft. All procedural deaths occurred in patients who received IdAc more than a year from diagnosis. Nine further patients died, eight following progression of CML. 72% of transplanted patients showed a major cytogenetic response but most cases have returned to Philadelphia-positive haemopoiesis. 62% of autografted patients remain alive (median survival from diagnosis 52 months). Four of the 11 patients who did not receive a transplant remain in chronic phase.     

Flow cytometric detection of cytomegalovirus antigen in peripheral blood cells after bone marrow transplantation

We report 13 normal peripheral blood stem cell (PBSC) donors who had a second PBSC collection for allogeneic transplantation performed after the first. The median interval between the first and second collection was 5 months. Mobilization was achieved with filgrastim (12 μg/kg/d). No significant difference was found in the median pre-apheresis leucocyte count (×10⁹/l) between the two donations (40.2 v 38.5; P  = 0.91). The median apheresis yield (×10⁶ CD34⁺ cells/litre blood processed, first apheresis) was also similar (28 v 27.3; P  = 0.91). Filgrastim-related adverse events were comparable. These data suggest that second PBSC collections are feasible, similarly tolerated and provide comparable apheresis yields.     

Role of antithymocyte globulin in matched sibling donor peripheral blood stem cell transplantation for hematologic malignancies

Background:High incidence of chronic graft-versus-host disease (GVHD) has been a major drawback of matched sibling donor peripheral blood stem cell transplantation (MSD -PBSCT). This study aimed to investigate the safety and efficacy of antithymocyte globulin (ATG) as a standardized part of GVHD prophylaxis in patients receiving MSD -PBSCT.Methods:A total of 72 patients with hematological malignancies receiving MSD -PBSCT who displayed similar baseline characteristics were either given rabbit ATG ( n = 42) or no ATG (n = 30), in addition to cyclosporine, methotrexate, and mycophenolate mofetil as a standard GVHD prophylaxis regimen. Either patients or donors aged ≥40 years were included in the study. Thymoglobulin was administered at a daily dose of 1.5 mg/kg on day −5 and 3.5 mg/kg on day −4 prior to transplant (the total dose was 5 mg/kg)Results:After a median follow-up of 874 days, the 3-year cumulative incidence of chronic GVHD (cGVHD) was 37.3% in the ATG group and 52.1% in the non -ATG group. The 3-year overall and disease-free survival probability were 71.0% and 62.0% (ATG versus non -ATG, P = .262) and 66.7% and 58.4% (ATG versus non -ATG, P = .334). No difference was found in the 2-year cumulative incidence of nonrelapse mortality and relapse between the ATG and non -ATG groups. This significant reduction in the incidence of cGVHD without increased relapse risk and nonrelapse mortality led to a 3-year GVHD-free, relapse-free survival probability of 66.7% and 40.0% in the ATG and non-ATG groups, respectively.Conclusions:These data suggested that rabbit antithymocyte globulin in the current protocol for GVHD prophylaxis was well tolerable and efficacious.The clinical trial was registered on January 1, 2016 (ClinicalTrials.gov Identifier {"type":"clinical-trial","attrs":{"text":"NCT02677181","term_id":"NCT02677181"}}NCT02677181). https://clinicaltrials.gov/ct2/show/{"type":"clinical-trial","attrs":{"text":"NCT02677181","term_id":"NCT02677181"}}NCT02677181.     

Allogeneic bone marrow transplantation from unrelated donors using in vivo anti-T-cell globulin

Despite improvements in HLA typing, graft-versus-host disease (GVHD) continues to impair the results after volunteer unrelated donor bone marrow transplantation (VUD-BMT) in adult patients compared with matched sibling BMT. Here, the outcome after VUD-BMT using a specific regimen with high-dose anti-T-lymphocyte globulin (ATG) was analysed. Fifty-five adult patients, median age 34 years (range 17-55 years), with acute or chronic leukaemia or myelodysplastic syndrome (MDS) were transplanted in first complete remission (CR1)/first chronic phase (CP1) (early disease) (n = 21) or in advanced (CR2/CP2, no remission) disease (n = 34) from an unrelated marrow donor. GVHD prophylaxis consisted of ATG-S (Fresenius) 60-90 mg/kg b.w. prior to transplantation, in addition to cyclosporin A and short-course methotrexate. Graft failure did not occur and white blood cell count (WBC) > 1.0 x 10(9)/l was reached at median day +16. The cumulative incidence of acute (a)GVHD grade II-IV was 15% [95% CI (8%, 28%)] and of chronic GVHD was 51% [95% CI (38%, 68%)]. The cumulative incidence of relapse within 1 year was 0% [95% CI (0%, 19%)] and 21% [95% CI (11%, 40%)] for patients with early and advanced disease respectively. With a median follow-up of 28 months (range 16-45 months), 2-year disease-free and overall survival for patients transplanted in CR1/CP1 was 81% and 81% [95% CI (64%, 98%)], respectively, and for patients with advanced disease was 33% [95% CI (17%, 50%)] and 40% [95% CI (23%, 57%)] respectively. Complete and persistent donor chimaerism was seen in 77.5% of 40 patients evaluated. All 14 chronic myeloid leukaemia (CML)-CP1 patients became bcr-abl negative within 250 d. High-dose ATG pretransplant results in a low incidence of severe aGVHD without compromising donor chimaerism or elimination of minimal residual disease. Our results are similar to data obtained after matched sibling donor transplantation.     

G-CSF-mobilized peripheral blood progenitor cells for allogeneic transplantation: safety,kinetics of mobilization,and composition of the graft

Allogeneic transplantation of peripheral blood progenitor cells (PBPC) markes the general anaesthesia of the donor unnecessary and may result in more rapid engraftment and faster recovery of the immune system. We have studied G-CSF-mediated PBPC mobilization in healthy donors and analysed the cellular composition of the resulting PBPC grafts. PBPC grafts were obtained from nine healthy donors (18-67 years old) for allogeneic or syngeneic transplantation. Six donors received 10 μg/kg G-CSF per day, the others 5-6 μg/kg. Mobilization and harvesting were well tolerated except for moderate bone pain which occurred in all donors primed with 10 μg/kg. With 10 μg/kg, a 31-fold (9-62) enrichment of circulating CD34⁺ cells was observed with peak values constantly occurring on day 5 after the start of G-CSF administration. Starting harvest on day 5, one to three collections on consecutive days yielded 5.5 x 10⁶/kg (0.9-10.7) CD34⁺ cells, 219 x 10⁶/kg (106–314) T cells, and 34 x 10⁶/kg (23–67) NK cells per 10 litres leukapheresis volume. Altogether, PBPC grafts contained 3 times more CD34⁺ cells, 7 times more T cells, and 20 times more NK cells than five allogeneic marrow grafts that were analysed for comparison. The yield of CD34⁺ cells per 10 litres apheresis volume as well as the height of the CD34⁺ peak in peripheral blood were inversely correlated to the age of the donor. In the donors primed with 5–6μg/kg G-CSF the increase of circulating CD34⁺ cells (4–7-fold enrichment) and the CD34⁺ cell yield per 10 litres leukapheresis volume (1 x 10⁶/kg [0.8-2-2]) was much smaller compared with the 10μg/kg group. In conclusion, sufficient amounts of PBPC capable of restoring haemopoiesis in allogeneic recipients can be mobilized safely by administration of G-CSF (10 μg/kg s.c. for 5 d) in healthy donors, and harvested with one or two leukapheresis procedures. Whether the large numbers of T-cells and NK cells that are contained in the collection products may influence graft-versus-host and graft-versus-leukaemia reactivities of PBPC grafts remains to be determined.     

CD34~+ CD38~-细胞对异基因造血干细胞移植的影响研究

目的观察CD34+CD38-细胞对异基因造血干细胞移植术后造血重建和移植物抗宿主病(GVHD)的影响。方法分析2004年1月至2009年12月河南省人民医院血液科全相合异基因外周血干细胞造血干细胞移植78例,CD34+、CD34+CD38-细胞输入量与血缘全相合异基因外周血造血干细胞移植术后造血重建及GVHD发生率间的相关性。结果粒细胞、血小板恢复时间与CD34+CD38-细胞输入量呈负相关(r分别为-0.521、-0.448,P<0.01),与CD34+细胞输入量也呈负相关(r分别为-0.405、-0.371,P<0.05)。急性GVHD、慢性GVHD的发生与CD34+、CD34+CD38-、CD3+、CD4+、CD8+细胞输入量无相关性。结论输入高数量的CD34+CD38-细胞有利于移植术后的粒细胞、血小板快速恢复;对于预测术后造血恢复,CD34+CD38-细胞亚群输入量可能优于CD34+细胞总数。     

HLA不全相合外周血造血干细胞移植治疗急性白血病的临床观察

目的：探讨HLA不全相合外周血造血干细胞移植(PBSCT)治疗急性白血病(AL)的可行性。方法：应用HLA1个位点不合亲缘供体PBSCT治疗急性白血病2例，采用BUCY加ALG进行预处理，采用环孢素A、甲氨喋呤、霉酚酸酯和抗CD25单抗等联合预防移植物抗宿主病(GVHD)。结果：2例患者均获造血重建，并发Ⅱ度急性GVHD及局限性慢性GVHD。随访19个月均无病生存，恢复正常生活。结论：对缺乏HLA完全相合同胞供体的白血病患者，应用HLA不全相合的亲缘供体PBSCT治疗急性白血病具有一定的应用前景。     

Invasive fungal infections after allogeneic peripheral blood stem cell transplantation: incidence and risk factors in 395 patients

We have analysed the incidence and risk factors for the occurrence of invasive fungal infections (IFI) among 395 recipients of an allogeneic peripheral blood stem cell transplantation (PBSCT) from a human leucocyte antigen (HLA)-identical sibling. IFI (n = 50) occurred in 46 patients, giving an overall probability of 14%. There were 12 cases of invasive candidiasis (3%), with only one death. Non-Candida IFI occurred in 37 patients (12% probability), mostly invasive aspergillosis (n = 32). In multivariate analysis the only two significant variables associated with a higher risk of developing a non-Candida IFI were the development of moderate-to-severe graft-versus-host disease (GvHD, P < 0.0001; OR 4.6) and having received steroid prophylaxis for GvHD (P = 0.04; OR 2.1). In multivariate analysis the variables associated with a lower overall survival after PBSCT were development of a non-Candida IFI (P < 0.0001; OR 5.6), non-early disease phase (P = 0.0001; OR 1.9), steroid prophylaxis (P = 0.02; OR 1.4), moderate-to-severe GvHD (P = 0.01; OR 1.6) and cytomegalovirus infection post transplant (P = 0.001; OR 1.8). Our results show that non-Candida IFI (in particular aspergillosis) was an important cause of infectious morbidity and mortality after an HLA-identical sibling PBSCT, while invasive candidiasis was rare. Use of steroid prophylaxis and, in particular, the development of moderate-to-severe GvHD post transplant were risk factors for non-Candida IFI. Prophylactic strategies for these infections should thus take into account these risk factors.     

家兔自体外周血干细胞移植治疗下肢缺血的实验研究

目的探索自体外周血干细胞移植治疗下肢动脉缺血性疾病的可行性。方法于2005-01～2005-04进行家兔自体外周血干细胞移植治疗下肢缺血实验研究。建立家兔双后肢缺血动物模型,取经动员的自体外周血干细胞制备成干细胞悬液,注射于右后肢缺血部位,分别于第2、4周后行数字减影血管造影术(DSA),取双侧后肢内收肌和腓肠肌标本,病理切片观察血管新生程度,CD31免疫组化检测毛细血管密度。结果DSA动脉造影及病理切片显示自体外周血干细胞移植的家兔缺血右后肢较左侧肢动脉明显增多,CD31免疫组化标记毛细血管密度增加。结论自体外周血干细胞移植是一种方法简便、安全有效的治疗下肢缺血性疾病的方法。     

Allogeneic bone marrow vs. peripheral blood stem cell transplantation: a long-term retrospective single-center analysis in 329 patients

Objectives: Granulocyte colony‐stimulating factor‐mobilized peripheral blood hematopoietic stem cell transplantation (HSCT) provides a valuable and increasingly used alternative to bone marrow transplantation (BMT). This retrospective study aimed at determining whether the stem cell source is predictive for outcome, relapse incidence, non‐relapse mortality, and severity and incidence of both, acute and chronic graft‐versus‐host disease (GVHD) in patients undergoing allogeneic HSCT. Patients and methods: Between 1983 and 2007, 329 adult patients (median age 40, range 18–76) received a first allogeneic HSCT from either sibling (n = 203) or volunteer unrelated donors (n = 126) at our institution. The source of stem cells was bone marrow in 177 (54%) and peripheral blood in the remaining 152 (46%) patients. Results: Overall survival was 37% (31–43%, 95% confidence interval, CI), the relapse incidence was 30% (25–36%, 95% CI), and the non‐relapse mortality was 43% (38–49%, 95% CI) for the entire cohort with no significant differences between peripheral blood stem cell or BMT. In patients receiving myeloablative conditioning, peripheral blood stem cell transplantation (PBSCT) was associated with a significantly lower non‐relapse mortality (32% vs. 46%, P = 0.05), which, however, was restricted to standard‐risk disease (23% vs. 42%, P = 0.02). The overall cumulative incidences of acute GVHD II–IV were 51% and 54% following bone marrow and PBSCT, respectively. Severe acute GVHD III–IV was significantly more frequent after BMT (24% vs. 14%, P = 0.04), whereas chronic GVHD was significantly more frequent following PBSCT (48% vs. 24%, P = 0.0001). By multivariate analysis, PBSCT was only predictive for chronic GVHD (RR 2.29, P = 0.02). Conclusion: Although we failed to demonstrate any advantage of PBSCT over conventional BMT with regard to overall survival, relapse incidence and non‐relapse mortality PBSCT were associated with a significantly higher incidence of chronic graft‐versus‐host disease. Therefore, and by virtue of observations, that some patient groups might benefit from either stem cell source, there is still need for prospective randomized trials with special emphasize on quality of life in long‐term survivors.     

异基因外周血干细胞移植治疗白血病的临床研究

目的　探讨异基因外周血干细胞移植（allo-PBSCT）的植活情况,移植物抗宿主病（GVHD）的发生率和临床结果。方法　回顾性分析1997年6月至1999年5月在我院实行的40例allo-PBSCT的临床资料。以粒细胞集落刺激因子(G-CSF)5μg/kg,12h1次,皮下注射动员外周血造血干细胞;输入CD     

单个核细胞计数对异基因外周血干细胞移植后造血重建的预测研究

目的探讨单个核细胞(MNC)计数作为造血干(祖)细胞含量的独立指标预测异基因外周血干细胞移植(allo-PBSCT)后造血重建的可行性。方法将暨南大学附属第一医院血液科2000年1月至2008年12月120例allo-PBSCT患者分为MNC组(83例)和CD34+细胞组(37例),MNC组以≥4×108/kg为采集目标,CD34+细胞组以≥4×106/kgCD34+细胞为采集目标。比较两种计数指标对造血重建和供者采集次数的影响,并分析不同MNC剂量对造血重建的影响。结果MNC组受者输入MNC的中位数为6.81×108/kg,CD34+细胞组受者输入CD34+细胞的中位数为5.05×106/kg;两组造血重建率均为100%;两组中性粒细胞植活的中位时间均为移植后第11天(P0.05),血小板植活的中位时间均为移植后第12天(P0.05);两组供者1次采集率分别为100%和37.84%(P0.05);MNC组中HLA全相合与不全相合移植受者中性粒细胞植活的中位时间分别为移植后第11天和移植后第12天(P0.05),血小板植活的中位时间分别为移植后第12天和移植后第14天(P0.05);MNC剂量在(3～5.99)×108/kg递增时,剂量与造血重建呈正相关,而MNC剂量在达到6×108/kg后递增,则并未使植活时间随之进一步缩短。结论MNC计数单独作为造血干(祖)细胞含量的计数指标,不仅能可靠预示allo-PBSCT(包括HLA全相合与不全相合移植)后造血重建,其植活率和植活速度可与CD34+细胞相比拟,而且其供者1次采集率(100%)显著高于后者(37.84%),allo-PBSCT时MNC计数可取代CD34+细胞作为造血干(祖)细胞含量的独立指标。     

非低温保存自体外周造血干细胞移植14例报告

目的:探讨非低温保存外周造血干细胞的活性,为临床自体外周造血干细胞移植提供方便有效的保存方法。方法:常规方法动员外周造血干细胞,CS-3000PLUS血细胞分离机采集外周造血干细胞,将采集好的干细胞加入肝素钠1250U后,放置4℃冰箱保存48~72h,预处理结束24h后回输。结果:全部病例获造血重建,MNC、CD34 细胞回收率分别为95.7%、91.4%,与保存前相比差异无统计学意义(P>0.05)。结论:4℃冰箱保存外周造血干细胞48~72h的方法是安全、可靠的,适用于基层医院临床应用。     

The role of allogeneic stem cell transplantation in the management of acute myeloid leukaemia: a triumph of hope and experience

Acute myeloid leukaemia (AML) is the commonest indication for allogeneic stem cell transplantation (allo-SCT) worldwide. The accumulated experience of allografting in AML over the last four decades has provided critical insights into both the contribution of the conditioning regimen and the graft-versus-leukaemia effect to the curative potential of the most common form of immunotherapy utilised in standard clinical practice. Coupled with advances in donor availability and transplant technologies, this has resulted in allo-SCT becoming an important treatment modality for the majority of adults with high-risk AML. At the same time, advances in genomic classification, coupled with progress in the accurate quantification of measurable residual disease, have increased the precision with which allo-mandatory patients can be identified, whilst simultaneously permitting accurate identification of those patients who can be spared the toxicity of an allograft. Despite this progress, disease recurrence still remains a major cause of transplant failure and AML has served as a paradigm for the development of strategies to reduce the risk of relapse ‒ notably the novel concept of post-transplant maintenance, utilising pharmacological or cellular therapies.     

Mesenchymal stem cells obtained after bone marrow transplantation or peripheral blood stem cell transplantation originate from host tissue

Mesenchymal stem cells (MSC) obtained from human bone marrow have been described as adult stem cells with the ability of extensive self-renewal and clonal expansion, as well as the capacity to differentiate into various tissue types and to modulate the immune system. Some data indicate that leukapheresis products may also contain non-hematopoietic stem cells, as they occur in whole bone marrow transplantation (BMT). However, there is still controversy whether MSC expand in the host after transplantation like blood progenitor cells do. Therefore, we were interested in finding out if graft MSC can be detected in leukapheresis products and in bone marrow after BMT and peripheral blood stem cell transplantation (PBSCT). Every sample from total bone marrow transplants exhibited growth of MSC after in vitro culture, but not one of nine leukapheresis products did. In addition, bone marrow aspirates of 9 patients receiving BMT and of 18 patients after PBSCT were examined for origin of MSC. Almost all MSC samples exhibited a complete host profile, whereas peripheral blood cells were of donor origin. We conclude that even if trace amounts of MSC are co-transplanted during PBSCT or BMT, they do not expand significantly in the host bone marrow.     

Transplantation of peripheral blood progenitor cells from HLA-identical sibling donors

Transplantation of peripheral blood progenitor cells (PBPCs) has largely replaced autologous bone marrow transplantation. The same might occur in the allogeneic setting if the favourable initial experience with allogeneic PBPCT is confirmed. We analysed all primary transplants utilizing unmodified PBPC from HLA-identical sibling donors reported to the European Group for Blood and Marrow Transplantation (EBMT) for 1994. 59 patients with a median age of 39 years received myeloablative therapy for acute myelogenous leukaemia (23 patients), acute lymphoblastic leukaemia (13), chronic myelogenous leukaemia (nine), lymphoma (seven), or other diagnoses (seven) mostly of advanced stages followed by transplantation of allogeneic PBPC. Three patients died soon after grafting, the others showed prompt haemopoietic recovery with median times to recover an absolute neutrophil count (ANC) above 0.5 and 1.0×10⁹/l of 15 (range 9–27) and 17 d (range 10–28) respectively. Time to platelet recovery above 20 or 50×10⁹/l was 16 (range 9–76) and 18 d (range 12–100) respectively. 27 patients (46%) developed no or mild acute graft-versus-host disease (GVHD). The incidence of moderate (grade II) disease was 27%; 24% of the patients developed severe acute GVHD (grades III or IV). 55% of patients who were alive 90 d after transplantation developed chronic GVHD, the probability to develop extensive chronic GVHD was 32% (95% confidence interval 22–42) with a median follow-up of 14 months. Overall and event-free survival (EFS) at 1 year were 54% (CI 48–60) and 50% (CI 43–57), respectively, the relapse incidence was 23% (CI 17–29). EFS was 67% (CI 55–79) in patients transplanted for acute leukaemias in first complete remission, chronic myelogenous leukaemia in first chronic phase, or severe aplastic anaemia. Transplantation of allogeneic PBPC resulted in prompt and durable engraftment. The incidence and severity of acute and chronic GVHD seemed comparable to that observed after allogeneic BMT. Overall and event-free survival in this cohort of patients, most of whom suffered from advanced leukaemia or lymphoma, is encouraging, suggesting that the high numbers of T lymphocytes and/or natural killer cells contained in a typical PBPC collection product exert a vigorous graft-versus-leukaemia effect. Further evaluation of allogeneic PBPCT is highly desirable.     

ABO血型不合对外周血造血干细胞移植GVHD影响的Meta分析

目的:移植物抗宿主病(GVHD)是异基因造血干细胞移植(allo-HSCT)中常见的并发症,其与ABO血型不合的关系尚存在争议,本研究利用Meta分析评价ABO血型不合对外周血造血干细胞移植GVHD的影响。方法:使用关键词检索PubMed,筛选出与本研究目的相符的文献,采用Review Manager 5.0统计软件进行分析。结果:初筛65篇文献,最终纳入6篇文献共计884例,其中血型不合417例,血型相合467例。血型不合组(包括主侧不合、次侧不合和主次侧均不合)发生aGVHD的OR=1.53(P=0.004),仅计算次侧不合组OR=1.40(P=0.14),cGVHD在血型相合与不合组之间无统计学差异。结论:Meta分析支持ABO血型不合提高外周血造血干细胞移植aGVHD的风险,但对cGVHD无影响。清髓方案可能促进ABO血型不合移植受者发生aGVHD。