Peripheral Blood Hematopoietic Stem Cell Research Articles

Circulating hematopoietic progenitors and CD34+ cells predicted successful hematopoietic stem cell harvest in myeloma and lymphoma patients: experiences from a single institution

BackgroundPrevious studies have shown that the numbers of both circulating hematopoietic progenitor cell (HPC) and CD34+ cell are positively correlated with CD34+ cell harvest yield. However, the minimal numbers of both circulating HPCs and CD34+ cells required for performing an efficient hematopoietic stem cell (HSC) harvest in lymphoma and myeloma patients have not been defined in our institution.Patients and methodsMedical records of 50 lymphoma and myeloma patients undergoing peripheral blood HSC harvest in our institution were retrospectively reviewed. The minimal and optimal HSC harvest yield required for the treatment was considered to be ≥2×106 CD34+ cells/kg and ≥5×106 CD34+ cells/kg, respectively.ResultsThe minimally required or optimal HSC yield obtained was not influenced by age (≥60 years), sex, underlying malignancies, disease status, multiple rounds of chemotherapy, or history of radiotherapy. The numbers of both circulating HPC and CD34+ cell were higher in patients with minimally required HSC yields (P=0.000 for HPC and P=0.000 for CD34+ cell) and also in patients with optimal HSC yields (P=0.011 for HPC and P=0.006 for CD34+ cell). The cell count cutoff for obtaining minimally required HSC harvest was determined to be 20/mm3 for HPCs and 10/mm3 for CD34+ cells. Furthermore, the cell count cutoff for obtaining optimal HSC harvest was determined to be 60/mm3 for HPCs and 35/mm3 for CD34+ cells.ConclusionA total of 60/mm3 of HPCs and 35/mm3 of CD34+ cells in peripheral blood predicted optimal HSC harvest in lymphoma and myeloma patients.

Exercise as an Adjuvant Therapy for Hematopoietic Stem Cell Mobilization.

Hematopoietic stem cell transplant (HSCT) using mobilized peripheral blood hematopoietic stem cells (HSPCs) is the only curative strategy for many patients suffering from hematological malignancies. HSPC collection protocols rely on pharmacological agents to mobilize HSPCs to peripheral blood. Limitations including variable donor responses and long dosing protocols merit further investigations into adjuvant therapies to enhance the efficiency of HSPCs collection. Exercise, a safe and feasible intervention in patients undergoing HSCT, has been previously shown to robustly stimulate HSPC mobilization from the bone marrow. Exercise-induced HSPC mobilization is transient limiting its current clinical potential. Thus, a deeper investigation of the mechanisms responsible for exercise-induced HSPC mobilization and the factors responsible for removal of HSPCs from circulation following exercise is warranted. The present review will describe current research on exercise and HSPC mobilization, outline the potential mechanisms responsible for exercise-induced HSPC mobilization, and highlight potential sites for HSPC homing following exercise. We also outline current barriers to the implementation of exercise as an adjuvant therapy for HSPC mobilization and suggest potential strategies to overcome these barriers.

Comparable Outcome of Allogeneic versus Autologous Hematopoietic Peripheral Blood Stem Cell Transplantation in Acute Myeloid Leukemia Patients with Normal Karyotype and FLT3-ITD Negative

Introduction: Optimal post-remission treatment for acute myeloid leukemia patients with normal karyotype (AMLNK) in first complete remission (CR1) who lacks an HLA identical donor is still not well-defined. Aim of the Work: To compare the outcome of allogeneic versus autologous peripheral blood stem cell transplantation (PBSCT) in adult AML patients regarding toxicities of transplant procedure, transplant-related mortality (TRM), disease free survival (DFS) and overall survival (OS). Patients and Methods: 43 AML patients were included; 34 patients (with a median age 28 years) received myeloablative allogeneic PBSCT from a matched sibling donor while 9 patients (with a median age 36 years) received PBSC autograft. All patients had a normal karyotype (NK), FMS-like tyrosine kinase 3 internal tandem duplication (FLT3 ITD) negative and were in CR1. Results: After a median follow up of 21.5 months (0.3- 46.5), the cumulative 2-year OS and DFS in the allogeneic group were 73.5% and 70.6% respectively, compared to 74.1% and 64.8%, respectively in the autologous group (p=0.690 and 0.768). Increasing number of consolidation cycles (>3) and lower CD34 stem cell dose were associated with lower relapse rates and higher DFS in the autologous group. Conclusion: Preliminary data show a comparable outcome of autologous compared to allogeneic PBSCT in patients with AML-NK and FLT3 ITD negative in CR1. In absence of matched sibling donor, autologous PBSCT may provide an acceptable post remission therapy for patients with low risk molecular profile.

The high effect of chemomobilization with high-dose topside and G-CSF in autologous hematopoietic peripheral blood stem cell transplantation (Single centre experience)

The high effect of chemomobilization with high-dose topside and G-CSF in autologous hematopoietic peripheral blood stem cell transplantation (Single centre experience)

Outcome Comparison of Lymphoma and Myeloma Patients after Autologous Stem Cell Transplantation (ASCT) with Peripheral Blood Stem Cell Mobilization Between Plerixafor (P) Mobilized in Poor Mobilizer Patients and Non-Plerixafor Mobilized Patients

Background: P is used in combination with G-CSF to improve the mobilization of peripheral blood hematopoietic stem cells in poor mobilizers. Limited data are available in regard to effects of P on post-transplant outcomes in comparison with non-P chemomobilized patients.Methods: In this retrospective study, we compare the engraftment and the outcomes of 34 patients mobilized with P + G-CSF or just-in-time rescue P in combination with chemotherapy and G-CSF to 143 patients (control group) mobilized with G-CSF with or without chemotherapy in lymphoma and myeloma patients who underwent ASCT between February 2012 and April 2014 at the University of Maryland Greenebaum Cancer Center. Post-transplantation outcomes including infections, hematologic recovery, relapse, progression and survival were recorded.Results: The median number of collected of CD34+ cells/Kg was 5.9 x 10(6) in the P group and 12.3 x 10(6) in the control group (p=0.0002). Median time to neutrophil engraftment (>0.5 × 10(9) /L) was comparable between the 2 groups: 12 days for the P group and 11 for the control group (p=0.28). There was a trend toward a shorter time to platelet engraftment (>20 × 10(9) /L) in the control (12 days) compared to the P group (14 days) (p=0.056). Progression-free survival at 1 year after (ASCT) was 88.2% in the P group and 81.8% in the control group. There was no difference in the overall survival of both groups (p=0.62)Conclusions: Short and long-term engraftment and outcomes after ASCT seem to be comparable in lymphoma and myeloma patients receiving plerixafor compared to chemomobilized patients without plerixafor. This observation support the use of plerixafor + G-CSF or just-in-time rescue P in patients who mobilize poorly without P. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

A Pilot Study of Peripheral Blood Hematopoietic Stem Cells Mobilization with the Combination of Bortezomib and G-CSF in Multiple Myeloma and Non-Hodgkin's Lymphoma Patients.

Background: Bortezomib has become an integral part of front-line therapy of multiple myeloma in a large majority of patients. There are preliminary reports which show that addition of bortezomib can augment the peripheral blood CD34 count during stem cell mobilization. In this single center prospective trial we added bortezomib to G-CSF to evaluate the effects of bortezomib on peripheral CD34 counts and collection.Methods: Patients aged 18-70 years with diagnosis of multiple myeloma (MM) or non-hodgkin's lymphoma (NHL) who were eligible for autologous stem cell transplantation (ASCT) and had received no more than three prior chemotherapeutic regimens were eligible for the study. Patients were enrolled in two groups. Group A (N=3) received G-CSF 16mcg/kg for 5 days and proceeded to stem cell collection on D5 and then received bortezomib 1.3mg/m2 on D5 after stem cell collection and G-CSF 16mcg/kg on D6, 7, 8 and repeat stem cell collection on D6, 7, 8 till the goal was achieved. Group B (N=17) received G-CSF 16mg/kg on D1-5 and received bortezomib 1.3mg/m2 on D4 and proceeded to stem cell collection on D5. If the patient was not able to collect the predefined goal CD34, G-CSF was continued on D 6, 7, 8 and a second dose of bortezomib 1.3mg/m2 was given on D7. Mobilization procedure was stopped once the predefined goal CD34 collection (4 x 106/kg for MM and 2 x 106/kg for NHL) had been collected.Primary objectives of the study was to determine if addition of bortezomib to G-CSF will result in an increase in PBSCs by > 2-fold and to achieve median neutrophil engraftment 12 days post ASCT. Secondary objectiveswere to evaluate the collected product for co-mobilization of lymphoma or myeloma cells and to determine if the use of bortezomib increases the mobilization of immune-stimulatory Dendritic cell (DC) -1 subsets.Results: A total of 23 patients were enrolled and 20 were evaluable for the results. Only one patient with NHL was enrolled and rest had MM. Median age of pts was 57 years, M/F 8/12, median number of previous chemotherapy regimens was 1 (range 1-3).The median peripheral blood CD34 count pre and post bortezomib in all patients were 28.8 x 106/kg and 37 x 106/kg respectively. All three patients in group A had drop in peripheral blood CD34 counts on D6 post bortezomib as they had undergone stem cell collection on day 5. In part B (N=17), 15 patients had increase in peripheral blood CD 34+ve cell counts with 4 patients achieved doubling while 11 pts had less than doubling of peripheral blood CD34 count after receiving bortezomib. Two patients had minimal drop in the peripheral blood CD34 counts post bortezomib. Median number of CD34 cells collected in15 patients (part B) were 5.06 x 106 CD34 cells/kg (range 4-15.1). 18 patients proceeded to ASCT and median time to neutrophil engraftment (ANC ≥500/cumm) post transplant was 12 days (range 11-16) and platelet engraftment (Plt count ≥ 20,000/cumm) was 18 days (range 15-27). There was no significant change in DC1/DC2 ratio in both groups following treatment with bortezomib and G-CSF (Figure 1).In group A all three patients collected goal CD34 count on day 5 and 2/3 patients collected >4 x106 CD34 cells/kg on D6 post bortezomib and1/3 patients collected 2.6 x 106 on D6 post bortezomib. In group B (n=17), 2 patients were unable to collect because of low CD34 counts on D4 and D5, 11 pts collected the goal in one day (D 5) and 4 pts required two days of apheresis (D 5 and 6). None of the patients received D7 bortezomib.Conclusion: Use of bortezomib during autologous stem cell collection was safe and well tolerated. Majority of patients had increase in peripheral blood CD34 counts post bortezomib administration on D4. Future trials should explore bortezomib as an alternate strategy to chemo-mobilization in combination with growth factors. [Display omitted] [Display omitted] DisclosuresOff Label Use: Bortezomib for stem cell mobilization. Lum:Karyopharm Therapeutics Inc: Equity Ownership; Transtarget.Inc: Equity Ownership. Deol:Bristol meyer squibb: Research Funding. Abidi:celgene: Speakers Bureau; Millenium: Research Funding.

Clinical Analysis of 4 AML Patients Aged Over 80 Years Treated with Chemotherapy Combined with Haploidentical Hematopoietic Stem Cell Infusion

To investigate the efficacy and safety of MA (mitoxantrone and cytarabine) regimen chemotherapy combined with granulocyte-colony stimulating factor (G-CSF)-mobilized family related HLA-haploidentical donor peripheral blood hematopoietic stem cell (G-PBHSC) infusion for the treatment of acute myeloid leukemia (AML) patients aged over 80 years. Four elderly patients with AML were treated in Chinese Second Artillery General Hospital from August 2008 to September 2013. The proportion of male to female was 1 : 3 and the median age 83 (80-85) years. All patients received programmed infusions of G-PBHSC after MA regimen chemotherapy without graft-versus-host disease (GVHD) prophylaxis. After complete remission (CR), patients only received G-PBHSC infusion without chemotherapy. Three cases achieved CR and their disease free survival (DFS) time was 18, 8, 6 months, respectively. 1 case did not reach remission after 2 cycles chemotherapy. The median overall survival (OS) time was 10 (3-20) months. No GVHD was observed in any of the patients during treatment. Concludsion: The combination of chemotherapy and programmed haploidentical G-PBHSC infusion is an alternative approach for AML patients aged over 80 years.

High efficiency of peripheral blood hematopoietic stem cell mobilization in multiple myeloma: one case report and literature review

Objective To explore the influential factors of obtaining high proportion of CD34+ cells in patients with multiple myeloma (MM) peripheral blood hematopoietic stem cell (HSC) mobilization. Methods One patient with MM who was 64 years old and hospitalized in Department of Hematology, Hainan Branch of Chinese PLA General Hospital was enrolled into this study on January 2014. The patient achieved complete remission after four courses of bortezomib + doxorubicin hydrochloride liposome + dexamethasone induction regimens. Then peripheral blood HSC was mobilized with bortezomib + high dose cyclophosphamide + dexamethasone regimens in combination with recombinant human granulocyte colony-stimulating factor (rhG-CSF) and recombinant human thrombopoietin (rhTPO). Mobilization effect was reported with its relevant literature reviewed. Results The proportion of CD34+ cells in HSC graft was 6.80%. The number of mononuclear cells and CD34+ cells in harvest of single collection was 3.17×108/kg and 19.20×106/kg, respectively. Engraftment time of neutrophil and platelet was 10 days and 12 days after HSC transplantation, respectively. Conclusions Chemotherapy in combination with rhG-CSF and rhTPO is effective and safety for autologous HSC mobilization in MM patient. As this research is just a case report, it still needs many large sample and multi-center randomized controlled trials to confirm whether it can be widely used in MM patients. Key words: Multiple myeloma; Hematopoietic stem cell transplantation; Hematopoietic stem cell mobilization

Biosimilar Filgrastim (Tevagrastim, XMO2) for AllogeneicHematopoietic Stem Cell Mobilization andTransplantation in Patients with Acute MyelogenousLeukemia/Myelodysplastic Syndromes.

Human recombinant granulocyte colony-stimulating factor (G-CSF), filgrastim (Neupogen; Amgen, Thousand Oaks, CA, USA), has been widely used for the mobilization of CD34(+) hematopoietic stem cells (HSC) from healthy donors. The experience with biosimilar G-CSF agents in this area is limited. We performed a prospective study assessing Tevagrastim (biosimilar filgrastim, XMO2; Teva, Israel) for mobilization of CD34(+) peripheral blood HSC in HLA-matched healthy sibling donors for transplantation in 24 patients with acute myelogenous leukemia (AML) and high-risk myelodysplastic syndromes (MDS) (NCT01542944). Results were compared to a historical control group of sibling donors who received filgrastim for stem cell mobilization for allogeneic stem cell transplantations in patients with AML and MDS. The healthy donors received Tevagrastim or filgrastim in a dose of 10 μg/kg body weight (BW) subcutaneously for 4 days. The target yields of CD34(+) cells was 5× 10(6) CD34(+) cells/kg BW of the recipient. A median 10.2× 10(6) (range, 2.52 to 35.4) and 9.35× 10(6) (range, 3.7 to 30.6) CD34(+) cells per kg BW were collected in the Tevagrastim and filgrastim groups, respectively. All patients promptly engrafted with a median day of absolute neutrophil count (ANC) of >.5× 10(9)/L and >1× 10(9)/L of 13 days (range, 10 to 21) and 13.5 days (range, 10 to 22), respectively in the Tevagrastim group and 12 days (range, 10 to 18) and 13 days (range, 10 to 18) in the filgrastim group, respectively. Platelets reached counts of >20× 10(9)/L and >50× 10(9)/L within a median of 14 days (range, 11 to 33) and 17 days (range, 12 to 33) in the Tevagrastim group and 13 (range, 10 to 29) and 15 (range, 10 to 32) days in the filgrastim group, respectively. The donors developed only mild and transient side effects, which were not different between the Tevagrastim study group and the filgrastim historical control group. Similarly, the transplantation-related toxicitiesand outcomes did not differ between the patients who underwent transplantation with Tevagrastim-mobilized grafts and the filgrastim historical controls. In summary, we observed a similar yield of CD34(+) stem cell mobilization in the Tevagrastim study group and the filgrastim historical control group with similar engraftment kinetic, hematopoietic reconstitution, and transplantation outcome. Tevagrastim administration was safe with minimal side effects and toxicity not different than historical controls. The lack of significant differences for all parameters of stem cell collection, engraftment, and safety with the biosimilar XMO2 Tevagrastim demonstrate the "similarity" of the biosimilar and recombinant human G-CSF inthis indication.

Clinical neurorestorative progress in amyotrophic lateral sclerosis

Clinical neurorestorative progress in amyotrophic lateral sclerosis Lin Chen,1,2,5 Hongyun Huang,3 Haitao Xi,4,5 Gengsheng Mao3 1Medical Center, Tsinghua University, 2Tsinghua University Yuquan Hospital, 3General Hospital of Chinese People's Armed Police Forces, 4Beijing Rehabilitation Hospital of Capital Medical University, 5Beijing Hongtianji Neuroscience Academy, Beijing, People’s Republic of China Abstract: Amyotrophic lateral sclerosis is a progressive and fatal neurodegenerative disease characterized by progressive paralysis and motor neuron death. In addition to symptomatic managements such as ventilation and nutritional support, neurorestorative therapies have demonstrated anti-neurodegenerative potential and may improve quality of life for patients. Currently, clinical neurorestorative strategies include pharmacological management (granulocyte colony stimulating factor), neuromodulatory intervention (repetitive transcranial magnetic and cortical stimulation), cell transplantation (bone marrow stromal cells, olfactory ensheathing cells, granulocyte colony stimulating factor-mobilized peripheral blood stromal cells, hematopoietic stem and progenitor cells, neural stem/progenitor cells, CD133+ cells and CD34+ cells), bioengineering and tissue engineering therapy, and combined neurorehabilitative treatment. In this review, we describe the latest progress in clinical neurorestorative management of amyotrophic lateral sclerosis and discuss the underlying evidence base. Keywords: amyotrophic lateral sclerosis, neurorestorative treatment, cell transplantation, clinical trial

19. Augment, revival and repair

It is well known that decreased temperature results in the suppression of metabolic activity and, thus, in a reduction of the rate at which deterioration of an unnourished biological system would occur. The freezing process, however, is not necessarily benign; it generally induces extreme variations in chemical, thermal, and electrical properties that could be expected to alter cellular membranes, intracellular organelles, and the associated delicate intracellular system functions. Given the extreme complexity of even the simplest biological cells, it is therefore remarkable that a reversible state of suspended animation by freezing is possible at all. Biological responses to cold stress can include (among others) disassembly of cell cytoskeleton, apoptosis, osmotic stress, oxidative stress and attenuation of transcription and translation. As a result, surviving post thawed cells may not function immediately as they did before, or may have been completely compromised. As deeper understanding of the molecular mechanisms has been uncovered, issues affecting the transcriptome due to cold stress have become identifiable as areas requiring attention. While the goal of any preservation strategy should be geared towards outcomes with no change to the cell or biological system (e.g. tissue or organ) pre-preservation, the ability to repair cells post preservation could mitigate inevitable compromises. Additionally, arbitrary “viability” counts generally accepted by various regulatory bodies need to be examined from the standpoint of evidence. Historically, some situations, such as autologous peripheral blood hematopoietic stem cell transplantation, have required higher doses of cells with lower viability due to the nature of preserving that system. Outcomes from these transplants can still yield good results in vivo. Preservation protocols that combine optimized preservation, “adjuvants” and factors that can provide support in vivo might be key to solving the very complex situations in organ preservation and importantly, use.

Circulating hematopoietic stem cells and putative intestinal stem cells in coeliac disease.

BackgroundThe intestinal stem cells (ISC) modulation and the role of circulating hematopoietic stem cells (HSC) in coeliac disease (CD) are poorly understood. Our aim was to investigate the longitudinal modifications in peripheral blood HSC traffic and putative ISC density induced by gluten-free diet (GFD) in CD.MethodsThirty-one CD patients and 7 controls were enrolled. Circulating CD133+ and CD34+ HSC were measured by flow cytometry, at enrolment and after 7 days and 1, 3, 6, 12, and 24 months of GFD. Endoscopy was performed at diagnosis and repeated at 6, 12, and 24 months following GFD. We used the Marsh-Oberhuber score to evaluate the histological severity of duodenal damage; immunohistochemistry was employed to measure the intraepithelial lymphoid infiltrate (IEL, CD3+ lymphoid cells) and the putative ISC compartment (CD133+ and Lgr5+ epithelial cells).ResultsAt enrolment, circulating HSCs were significantly increased in CD patients and they further augmented during the first week of GFD, but progressively decreased afterwards. CD patients presented with villous atrophy, abundant IEL and rare ISC residing at the crypt base. Upon GFD, IEL progressively decreased, while ISC density increased, peaking at 12 months. After 24 months of GFD, all patients were asymptomatic and their duodenal mucosa was macroscopically and histologically normal.ConclusionsIn active CD patients, the ISC niche is depleted and there is an increased traffic of circulating HSC versus non-coeliac subjects. GFD induces a precocious mobilization of circulating HSC, which is followed by the expansion of the local ISC compartment, leading to mucosal healing and clinical remission.

Infused autograft lymphocyte-to-monocyte ratio and survival in T-cell lymphoma post-autologous peripheral blood hematopoietic stem cell transplantation.

BackgroundThe infused autograft lymphocyte-to-monocyte ratio (A-LMR) is a prognostic factor for survival in B-cell lymphomas post-autologous peripheral hematopoietic stem cell transplantation (APHSCT). Thus, we set out to investigate if the A-LMR is also a prognostic factor for survival post-APHSCT in T-cell lymphomas.MethodsFrom 1998 to 2014, 109 T-cell lymphoma patients that underwent APHSCT were studied. Receiver operating characteristic (ROC) and area under the curve (AUC) were used to identify the optimal cut-off value of A-LMR for survival analysis and k-fold cross-validation model to validate the A-LMR cut-off value. Univariate and multivariate Cox proportional hazard models were used to assess the prognostic discriminator power of A-LMR.ResultsROC and AUC identified an A-LMR ≥ 1 as the best cut-off value and was validated by k-fold cross-validation. Multivariate analysis showed A-LMR to be an independent prognostic factor for overall survival (OS) and progression-free survival (PFS). Patients with an A-LMR ≥ 1.0 experienced a superior OS and PFS versus patients with an A-LMR < 1.0 [median OS was not reached vs 17.9 months, 5-year OS rates of 87 % (95 % confidence interval (CI), 75–94 %) vs 26 % (95 % CI, 13–42 %), p < 0.0001; median PFS was not reached vs 11.9 months, 5-year PFS rates of 72 % (95 % CI, 58–83 %) vs 16 % (95 % CI, 6–32 %), p < 0.0001].ConclusionsA-LMR is also a prognostic factor for clinical outcomes in patients with T-cell lymphomas undergoing APHSCT.

Infused autograft lymphocyte to monocyte ratio and survival in T-cell lymphomas post-autologous peripheral blood hematopoietic stem cell transplantation.

e19500 Background: We recently reported that the infused autograft lymphocyte to monocyte ratio (A-LMR) is a prognostic factor for survival in B-cell lymphomas post-autologous peripheral hematopoietic stem cell transplantation (APHSCT). Thus, we set out to investigate if the A-LMR is also a prognostic factor for survival post-APHSCT in T-cell lymphomas. Methods: From 1998 to 2014, 109 T-cell lymphoma patients that underwent APHSCT were studied. Receiver operating characteristic and area under the curve identified an A-LMR ≥ 1 as the best cut-off value and validated by the k-fold cross-validation. Results: . Multivariate analysis showed A-LMR to be an independent prognostic factor for overall survival (OS) and progression-free survival (PFS). Patients with an A-LMR ≥ 1.0 experienced a superior OS and PFS versus patients with an A-LMR < 1.0 [median OS was not reached vs 17.9 months, 5-year OS rates of 87% (95 CI, 75%-94%) vs 26% (95 CI, 13%-42 %), p < 0.0001, respectively; median PFS was not reached vs 11.9...

A retrospective analysis of autologous peripheral blood hematopoietic stem cell mobilizations and collections in 149 multiple myeloma patients

目的了解多发性骨髓瘤（MM）患者自体外周血造血干细胞动员采集的状况及影响因素。方法回顾分析1998年1月至2014年3月149例MM患者采用环磷酰胺（CTX）或E-CHOP（依托泊苷+CTX＋表阿霉素+长春地辛＋泼尼松）化疗联合G-CSF动员采集外周血造血干细胞的资料[采集获得CD34+细胞数、成功（获得的CD34+细胞数≥2×106/kg）率、优良（获得的CD34+细胞数≥5×106/kg）率]。分析性别、年龄、疾病类型、DS分期、ISS分期、动员前治疗情况、动员时疾病状态、动员方案等因素与采集结果之间的关系。结果149例患者共进行了177例次动员采集，采集CD34+细胞中位数为3.20（0.13~22.34）×106/kg，采集成功率、优良率分别为74.5%、27.5%。单因素分析显示：性别、年龄（>60岁/≤60岁）、疾病类型、DS分期（Ⅲ/Ⅰ +Ⅱ）、ISS分期（Ⅲ/Ⅰ+Ⅱ）及动员方案（E-CHOP+G-CSF/CTX+ G-CSF）与采集获得CD34+细胞数、采集成功率无相关性（P>0.05），但>60岁的患者单次采集成功率较低（P<0.05），DS Ⅲ期的患者采集优良率较低（P<0.05）；动员前疗程数≤6、动员时疾病状态为部分缓解及以上的患者采集效果较好。动员前治疗情况影响干细胞采集成功率（P=0.006），疗程数>6的患者干细胞采集失败风险高（OR值为3.57, 95%CI 1.45~8.78）。结论性别、年龄、诊断类型、DS分期、ISS分期、动员采用的化疗方案对MM患者外周血干细胞采集成功率无影响，但年龄较大的患者需要两次动员。动员前疗程数较少、动员时相对稳定的疾病状态有利于造血干细胞的动员采集。

Hematopoietic reconstitution after transplantation of uncontrolled-rate cryopreservation autologous peripheral blood hematopoietic stem cells using -80 °C mechanical freezer

This study was to identify the efficacy of -80°C cryopreservated peripheral blood hemato-poietic stem cell (PBHSC) transplantation for hematopoietic reanstitution in patients. The efficacy of 104 patients underwent autologous peripheral blood hematopoietic stem cell transplantation using uncontrolled-rate freezing and storage at -80°C was evaluated. This cryopreservation method could effectively cryopreserve peripheral blood stem cells. Out of 104 patients only 2 patients died, other patients got hematologic reconstition satisfactorily, the median engrafement times of neutrophils and platelet were 12 and 14 days respectively, the activity of cells after rehabilitation was 94%, the mean recovery rates of CD34(+) cells and mononuclear cells (MNC) were 86% and 80.3% respectively. There were no significant influences on engrafement time in sex, chemotherapy circles and radiotherapy. The engrafement of leukocytes associated with amount of CD34(+) cells. This simple uncontrolled-rate freezing PBHSC at -80°C is safe, effective and economic, and can meet clinical needs. As compared with the classical cryopreservation, there were no significant differences in hematopoietic reconstitution. Therefore, this method worth to popularize and apply in clinic.

Adverse Events during Peripheral Blood Hematopoietic Stem Cell Mobilization in Light Chain Amyloidosis Patients

Adverse Events during Peripheral Blood Hematopoietic Stem Cell Mobilization in Light Chain Amyloidosis Patients

Engineered dendritic cells from cord blood and adult blood accelerate effector T cell immune reconstitution against HCMV

Human cytomegalovirus (HCMV) harmfully impacts survival after peripheral blood hematopoietic stem cell transplantation (PB-HSCT). Delayed immune reconstitution after cord blood (CB)-HSCT leads to even higher HCMV-related morbidity and mortality. Towards a feasible dendritic cell therapy to accelerate de novo immunity against HCMV, we validated a tricistronic integrase-defective lentiviral vector (coexpressing GM-CSF, IFN-α, and HCMV pp65 antigen) capable to directly induce self-differentiation of PB and CB monocytes into dendritic cells processing pp65 (“SmyleDCpp65”). In vitro, SmyleDCpp65 resisted HCMV infection, activated CD4+ and CD8+ T cells and expanded functional pp65-specific memory cytotoxic T lymphocytes (CTLs). CD34+ cells obtained from PB and CB were transplanted into irradiated NOD.Rag1−/−.IL2γc−/− mice. Donor-derived SmyleDCpp65 administration after PB-HSCT stimulated peripheral immune effects: lymph node remodeling, expansion of polyclonal effector memory CD8+ T cells in blood, spleen and bone marrow, and pp65-reactive CTL and IgG responses. SmyleDCpp65 administration after CB-HSCT significantly stimulated thymopoiesis. Expanded frequencies of CD4+/CD8+ T cell precursors containing increased levels of T-cell receptor excision circles in thymus correlated with peripheral expansion of effector memory CTL responses against pp65. The comparative in vivo modeling for PB and CB-HSCT provided dynamic and spatial information regarding human T and B cell reconstitution. In vivo potency supports future clinical development of SmyleDCpp65.

Loss of MXD3 induces apoptosis of Reh human precursor B acute lymphoblastic leukemia cells.

MXD3 is a transcription factor that plays an important role in proliferation of human DAOY medulloblastoma cells. Here, we demonstrate that MXD3 is highly enriched in human precursor B acute lymphoblastic leukemia (preB ALL) samples compared to mobilized peripheral blood mononuclear cells, bone marrow, or hematopoietic stem cells from healthy donors. MXD3 knock-down in the preB ALL cell line Reh resulted in decreased cell numbers with no change in G0/G1, S or G2/M populations but increased apoptosis compared to control cells. Our results suggest that MXD3 is important for survival of Reh preB ALL cells, possibly as an anti-apoptotic factor.

Reduced Relapse Risk in Acute Myelogenous Leukemia (AML) after Allogeneic Hematopoietic Stem Cell Transplantation (HCT) in Patients Who Experienced CMV Reactivation: Influence of Donor KIR Genotypes

Background: Recent studies demonstrate an intriguing association between CMV reactivation and reduced risk of post-transplant AML relapse, yet mechanistic explanations remain elusive. Donor killer immunoglobulin-like receptor (KIR) genotypes have been associated with both AML and CMV clinical outcomes post allogeneic HCT. In this study we evaluated the CMV reactivation association with AML relapse risk, and the possible impact of donor KIR genotypes on the CMV-AML relapse relationship.Patients/Methods: A consecutive case-series of 288 patients with AML who underwent allogeneic HCT between 1995 and 2011 at City of Hope were identified, patients were excluded who were not in remission at HCT, or received ex-vivo/in-vivo T cell depletion. A landmark analysis on 276 patients who survived without relapse for at least 55 days (this time point was selected given that the majority of CMV reactivations had occurred by this time, with very few relapse events). The median age was 49 years (range: 18 - 70). All patients were in 1st (n=229) or 2nd remission (n=47). Transplant conditioning included fully ablative (n=151) or reduced intensity regimens (fludarabine 125mg/m2 + melphalan 140mg/m2: n=125). Patients received peripheral blood hematopoietic stem cell (n=240) or bone marrow (n=36) grafts from matched sibling donors (n=190) or unrelated donors (8/8 match: n=86). Graft-versus-host disease prophylaxis consisted of a calcineurin inhibitor plus sirolimus or a short course of methotrexate. No patients in this cohort received prophylactic anti-CMV therapy. CMV was monitored by shell vial culture or polymerase chain reaction (PCR) once or twice a week from day +21 to day +100. Preemptive therapy with ganciclovir or foscarnet began when CMV reactivation was detected by positive culture or PCR. KIR genotyping was performed on donor DNA samples as previously described (Tissue Antigens, 2004;64:462) using 28 primers in 4 multiplex PCR reactions for detection of 14 functional KIR genes.Results: CMV reactivation was observed in 85 of 276 patients (31%). The 5-year cumulative incidence of relapse was 30.2% (95% CI: 24.1 – 37.7%) in patients without CMV reactivation, significantly higher than in patients experiencing CMV reactivation (12.1%: 95% CI 6.7 – 21.6%, p=0.002) (Figure 1). Other clinical variables showing a significant association or p<0.10 by univariate analysis were: cytogenetic risk (high risk vs others: 37.6% vs. 20.0%, p= 0.01), transplant era (before or after 2005: 18.6% vs 28.0%, p=0.065), and conditioning (flu-mel vs others: 30.0% vs. 20.3%, p=0.095). In multivariate analysis, only CMV reactivation and high-risk cytogenetics were highly significant (Table 1). The association between CMV reactivation and reduced AML relapse was independent of the donor/recipient CMV serostatus.We then examined the protective effect of CMV reactivation on relapse in subgroups of patients based on KIR genotypes (available in 251 cases). A significant protective effect on 5-yr relapse incidence (RI) was observed in the group with donor KIR B/X genotype (>1 activating KIR gene, n=164): 11.7% RI (95% CI 5.5 – 24.8) in patients with CMV reactivation vs. 29.5% (95% CI 22.0 – 39.6) in those without (p=0.016). No significant differences (p=0.3) in relapse were seen in the A/A genotype (only 1 activating KIR gene, n=87). Likewise, when we evaluated an activating KIR gene, 2DS1, which is reported to protect from AML relapse, the significant association was preserved in donors with 2DS1 (n=96): 11.5% RI (95% CI 4.0 – 33.4) in patients with CMV reactivation vs. 30.7% (95% CI 21.5 – 43.9) in those without (p=0.05). No significant relationship (p=0.12) between relapse and CMV reactivation was seen in donors without 2DS1 (n=155).Conclusions: Our data confirm the earlier observation of a significant association between CMV reactivation and protection from AML relapse, and suggest that in patients with donor-derived activating KIRs, HLA-KIR interactions may augment the protection from AML relapse after CMV reactivation. Table 1Landmark Analysis of Cumulative Incidence of Relapse Event = Relapse, Completing risk = non-relapse mortalityVariablesValue# of eventsHazard Ratio(95% CI)p-valueCMV ReactivationNo (n=191) Yes* (n=85)n = 56 n = 112.6 (1.4 - 4.9)0.003Cytogenetic RiskFav/Int/Unknown*(n=207) Unfavorable (n=69)n = 42 n = 251.8 (1.1 - 2.9)0.018* reference group, Fav - favorable, Int - intermediate [Display omitted] DisclosuresNo relevant conflicts of interest to declare.