Peripheral Blood Hematopoietic Stem Cell Research Articles

Clinical Analysis of Refractory Primary Central Nervouse System Lymphoma Treated with High-Dose MTX-Based Chemotherapy and Haploidentical Hematopoietic Stem Cell Infusion

To investigate the efficacy and safety of high-dose methotrexate-based chemotherapy combined with granulocyte-colony stimulating factor (G-CSF)-mobilized family related haploidentical donor peripheral blood hematopoietic stem cell (G-PBHSC) infusion for the treatment of patients with refractory primary central nervouse system lymphoma (PCNSL). Three patients with refractory PCNSL were treated in Department of Hematology of the General Hospital of the PLA's Rocket Force from March 2014 to September 2015. The sex ratio of male to female was 1:2 and the median age was 54(48-66)years old. All patients received programmed infusions of G-PBHSC after high-dose methotrexate-based chemotherapy without prophylaxis for graft-versus-host disease (GVHD). Three patients had received initial chemotherapy or radiotherapy after diagnosis, one patient achieved complete remission (CR) after 3 courses of treatment and remained in CR until the end of follow-up, 2 cases achieved partial remission (PR) and the progression-free survival (PFS) time was 10 and 7 months, respectively. The patients generally well-tolerated this therapy. The main adverse effects of patients were neutropenia, thrombocytopenia and infection related with chemotherapy after each course of treatment, the median recovery times of neutrophils and platelets were 11 and 12.5 days, respectively after of programmed infusions of G-PBHSC. No GVHD was observed in any of the patients during treatment. The combination of high-dose methotrexate-based chemotherapy with programmed haploidentical G-PBHSC infusion is a potential treatment alternative for refractory PCNSL patients.

Humanized Mouse Models for the Preclinical Assessment of Cancer Immunotherapy.

Immunotherapy is one of the most exciting recent breakthroughs in the field of cancer treatment. Many different approaches are being developed and a number have already gained regulatory approval or are under investigation in clinical trials. However, learning from the past, preclinical animal models often insufficiently reflect the physiological situation in humans, which subsequently causes treatment failures in clinical trials. Due to species-specific differences in most parts of the immune system, the transfer of knowledge from preclinical studies to clinical trials is eminently challenging. Human tumor cell line-based or patient-derived xenografts in immunocompromised mice have been successfully applied in the preclinical testing of cytotoxic or molecularly targeted agents, but naturally these systems lack the human immune system counterpart. The co-transplantation of human peripheral blood mononuclear cells or hematopoietic stem cells is employed to overcome this limitation. This review summarizes some important aspects of the different available tumor xenograft mouse models, their history, and their implementation in drug development and personalized therapy. Moreover, recent progress, opportunities and limitations of different humanized mouse models will be discussed.

Diagnosis and treatrnent of psoriasiform cutaneous chronic graft versus host disease and literatures review

Objective To explore the treatment and clinical therapeutic effect of psoriasiform cutaneous chronic graft versus host disease (cGVHD) in a patient with hybrid acute leukemia after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods On 9 April 2014, one case with hybrid acute leukemia who was treated with allo-HSCT in Institute of Hematology and Blood Disease Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, was selected as the research object. The conditioning regimen before allo-HSCT included idarubicin, cytosine arabinoside, busulfan, anti-thymocyte globulin and cyclophosphamide (CTX). The source of grafts was a combination of peripheral blood hematopoietic stem cells and bone marrow hematopoietic stem cells from human leukocyte antigen(HLA) haploidentical donor.The recipients received cyclosporin A, mycophenolate mofetil (MMF) and short-term methotrexate (MTX) for GVHD prophylaxis. Results Engraftment time was 17 d after allo-HSCT. Twenty-seven days and 7 months after allo-HSCT [34 d after donor lymphocyte infusion(DLI)], grade Ⅰ skin acute graft-versus-host disease (aGVHD) were occurred respectively, and both treated with methylprednisolone (MP) and cyclosporine A. One year after allo-HSCT, skin rash occurred on patient′s skin of the upper limbs, chest and abdomen. The patient was diagnosed with psoriasiform cutaneous cGVHD. After treatment with MP, CTX and cyclosporine A, the patient′s symptoms were relieved. Conclusions Hybrid acute leukemia patients with psoriasiform cutaneous cGVHD after allo-HSCT is rare. The usage of low-dose CTX combined with glucocorticoids and cyclosporine A can obtain better therapeutic effects. Key words: Hematopoietic stem cell transplantation; Graft vs host disease; Parapsoriasis; Leukemia, biphenotypic, acute; Skin diseases

Efficacy and safety evaluation of mobilization and collection of unrelated allogeneic peripheral blood stem cells

Objective To evaluate the efficacy and safety of the mobilization and collection of unrelated allogeneic peripheral blood stem cells. Methods The suitable stem cell mobilization plan was made in accordance with the hematopoietic stem cell mobilization plan of China Marrow Donor Program, the ruler of the hospital, and the donor's constitution. The unrelated allogeneic peripheral blood stem cells of 64 healthy donors were collected in the second Hospital of Shanxi Medical University from May 2012 to January 2017. The donor was infected one or several times with the mobilization agent granulocyte colony stimulating factor (G-CSF) by 5-10 μg·kg-1·d-1. After 3-4 days, peripheral blood hematopoietic stem cells were collected using COBE Spectra blood cell separator. Then, the effect and adverse reaction of donors were analyzed from different age and sex. Results It can achieve the acquisition requirements using 3 or 4 days of mobilization programs, mononuclear cells ≥ 5.0×108/kg, CD34+ cells ≥ 2.0×106/kg. The single acquisition success rate (the target acquisition of the number of mononuclear cells and CD34+) up to 65%, collection efficiency reached 52%, which could reduce the risk of donor and the cost of patients. The quality of donor stem cell of young was better than that of older persons. Sixteen donors (25%) had mild adverse reactions, and no special treatment was required. Conclusions Allogeneic stem cell mobilization is safe. Starting from save medical resources and the interests of the donor the 3 day or 4 days of mobilization scheme could improve the success rate of the single mobilization. During the collection process, the condition of donor hypocalcemia should be observed and health education should be given to relieve the tension of donor. Key words: Hematopoietic stem cell mobilization; Collection; Efficacy; Safety

Improved progression-free and event-free survival in myeloma patients undergoing PBSCH receiving a cyclophosphamide+G-CSF regimen than G-CSF alone.

Two regimens are commonly used for peripheral blood hematopoietic stem cell harvesting (PBSCH) in multiple myeloma: high-dose cyclophosphamide (HD-CY)+granulocyte-colony stimulating factor (G-CSF), and G-CSF alone. The objective of the present study was to evaluate the anti-myeloma effect of the PBSCH regimen including HD-CY. We retrospectively assessed harvesting efficiency, complications, and anti-myeloma effects in 115 patients receiving HD-CY+G-CSF (HD-CY group) and 32 patients receiving G-CSF alone (G-alone group). We collected>2×106 CD34-positive cells/kg from 93 and 75% of patients in the HD-CY and G-alone groups, respectively (P=0.0079). The mean HSC count was also higher in the HD-CY group. No severe complications were observed in the G-alone group, whereas 66% of patients in the HD-CY group were treated with intravenous antibiotics. The median progression-free and event-free survival (PFS and EFS) were longer in the HD-CY group than in the G-alone group (28 vs. 18months and 25 vs. 13months, respectively; P=0.0127 and 0.0139), with no difference in median overall survival. HD-CY showed anti-myeloma effect, as verified by prolonged EFS and PFS, when a vincristine, doxorubicin, and dexamethasone regimen was administered as induction before PBSCH.

The impact of preapheresis white blood cell count on autologous peripheral blood stem cell collection efficiency and HSC infusion side effect rate.

Autologous peripheral blood hematopoietic stem cell (PBSC) collection efficiency (CE) is reportedly affected by the patient's blood properties; however, studies to identify factors correlated with CE have shown inconsistent results. Additionally, variables such as stem cell graft granulocyte content and patient age, sex, and underlying disease, may be associated with hematopietic stem cell (HSC) infusion-related adverse reactions. In this study, we evaluated the correlation of preleukapheresis PB granulocyte count and PBSC harvest variables with CD34+ collection yield and efficiency, and thawed HSC infusion side effect occurrence. We evaluated data from 361 patients who had undergone autologous PBSC transplant. Large volume leukapheresis was the method for PBSC collection. Complete Blood Count and CD34+ cell enumeration were performed in the preapheresis PB and the apheresis product sample. The PBSC grafts were submitted to non-controlled rate freezing after addition of 5% DMSO plus 6% hidroxyethylstarch as a cryoprotectant solution. The cryopreserved graft was thawed in a 37°C water bath and then infused without further manipulation. The CD34+ yield was associated with preapheresis PB CD34+ count and immature granulocyte count. The PBSC CE was negatively correlated with preapheresis white blood cell (WBC), immature granulocyte and granulocyte count. The leukapheresis product total nucleated cell (TNC) and granulocyte content was correlated with the thawed graft infusion side effect occurrence. This study has shown that preapheresis PB WBC and granulocyte counts were associated with leukapheresis CE. Additionally, the leukapheresis product TNC and granulocyte content was correlated with thawed graft infusion side effect occurrence.

Successful T Cell Replete Haploidentical Peripheral Blood Hematopoietic Stem Cell Transplantation in a Young Girl with Diamond Blackfan Anemia

Successful T Cell Replete Haploidentical Peripheral Blood Hematopoietic Stem Cell Transplantation in a Young Girl with Diamond Blackfan Anemia

Factors Associated with Successful Mobilization and Collection of Peripheral Blood Hematopoietic Stem Cells in Autologous and Allogeneic Donors

BACKGROUND: Peripheral blood hematopoietic stem cells (PBSC) have largely replaced bone marrow derived stem cells in autologous transplantations, and have become the preferred source of stem cells in the majority of allogeneic transplantations. Sufficient number of mobilized and collected hematopoietic stem cells (HSC) is needed for successful hematopoietic stem cell transplantation.MATERIAL AND METHOD: This study was performed in the Institute for Transfusion Medicine of RM and the University Clinic of Hematology from 2008 till 2016. There were 30 allogeneic and 90 autologous donors that underwent mobilization and collection of PBSC. The association between possible predictive factors such as demographic characteristics, laboratory parameters and collection parameters in both groups, and mobilization strategy and clinical characteristics in autologous donors and number of collected PBSC was analyzed.RESULTS: There were 226 apheresis, 182 in autologous donors (mean 2, range 1-3) and 44 apheresis in 30 allogeneic donors (mean 1.5, range 1-2). The mean number of collected MNC in autologous donors was 3.09 x 108/kg and 2.85 x 106/kg CD34+ cells, and 3.23 x 108/kg MNC and 3.20 x 106/kg CD34+ cells in allogeneic donors. Significantly larger number of MNC and CD34+ cells was collected with the WBC set. There was a statistically significant correlation between the total number of collected MNC in autologous donors and platelet count before mobilization, the number of cycles in one apheresis procedure, quantity of collected graft and the number of collected MNC and CD34+ cells on the first day of harvestration. There was a statistically significant correlation between the total number of collected MNC in allogeneic donors and platelet count before mobilization, the number of cycles in one apheresis procedure, quantity of collected graft and number of MNC on first day of harvestration. There was a strong correlation between the number of collected MNC and CD34+ cells on the first harvest and the total number of collected MNC and CD34+ cells in poor mobilizers, and inverse correlation with the number of apheresis procedures. Donors who donated MNC ≤ 0.7 x 108/kg and/or ≤ 0.7 x 106/kg CD34+ cells on the first harvest (84.6%) were strong predictors of poor mobilizers.CONCLUSION: Determining the proper level of baseline and preaheresis laboratory parameters for initiating mobilization and apheresis procedure which is safe for donors and greatly efficient in collection of PBSC is needed for optimization of these procedures, as well as for early intervention in poor mobilizers.

Autograft immune effector cells and survival in autologous peripheral blood hematopoietic stem cell transplantation.

In addition to stem cells, T-cells, natural killer cells, dendritic cells, and monocytes are also collected and infused from the autograft in patients undergoing autologous peripheral blood hematopoietic stem cell transplantation. Recent reports have shown that these autograft immune effector cells can affect the clinical outcome postautologous peripheral blood hematopoietic stem cell transplantation. In this article, I will review the clinical impact on the survival of these autograft immune effector cells conferring the concept of autologous graft versus tumor effect.

Adverse Prognostic Factors for Morbidity and Mortality During Peripheral Blood Stem Cell Mobilization in Patients with Light Chain Amyloidosis

Patients with immunoglobulin light chain (AL) amyloidosis undergoing peripheral blood hematopoietic stem cell (PBSC) mobilization for autologous hematopoietic stem cell transplantation (auto-HCT) can experience significant morbidity and mortality. The purpose of this study was to characterize the adverse events and identify prognostic factors associated with the development of morbidity and mortality in patients with AL amyloidosis who had begun PBSC mobilization for auto-HCT. A retrospective study was performed in 101 consecutive patients with AL amyloidosis who underwent PBSC mobilization for auto-HCT between January 2006 and December 2013. A composite primary endpoint of morbidity and mortality during PBSC mobilization was used. Forty-one patients (41%) experienced at least 1 adverse event, including 4 deaths during PBSC mobilization. Adverse events included in this composite endpoint were cardiac events, thromboembolic events, bleeding events, unplanned hospitalization, weight gain >2% necessitating diuretic intervention, and death. Low serum albumin levels, elevated N-terminal pro-brain natriuretic peptide, and increased interventricular septal thickness were significantly associated with the composite primary endpoint (P = .024, .001, and .006, respectively). The median progression-free survival from the start of PBSC mobilization was 4.7 years, and the median overall survival was 6.5 years. In general, PBSC mobilization is associated with minimal complications, but patients with AL amyloidosis can experience more frequent and severe complications, such as volume overload and weight gain. Careful patient selection is warranted in patients with AL amyloidosis before proceeding to PBSC mobilization and auto-HCT.

DA-EPOCH Chemotherapy Combined with G-CSF Effectively Mobilizes Autologous PBHSC in NHL Patients

To study the effect of the DA-EPOCH chemotherapy combined with G-CSF and the CTX therapy with G-CSF on mobilizing and collecting the peripheral blood hematopoietic stem cells and the later hematopoietic recovery. Forty patients accepted mobilization and collection of peripheral blood stem cells(PBSC) after treated by CTX+G-CSF and DA-EPOCH+G-CSF therapy respectively, and were treated by auto-transfusion after BEAM pre-regimen. The mobilization efficacy, adverse effects and hematopoietic recovery after autologous transplantation were analyzed retrospectively. During the CTX+G-CSF mobilization, only one patient achieved the white blood cell(WBC) at 0.8×109/L, while the others were with the lowest WBC level above 2.0×109/L. The platelet counts were all normal with the exception of 3 cases at 80×109/L. The median percentage of CD34+ cells in one period of collection was 0.99(0.35-1.30)%. The median MNC was (3.80±2.05)×1010. The cumulative total of mononuclear cell was (5.84±2.48)×108/kg, and the median CD34+ cell count was 3.84(3.91-6.5)×106/kg. During the DA-EPOCH+G-CSF mobilization, the peripheral WBC count of patients were decreased to the lowest level at (0.2-1.4)×109/L. The platelet counts were all above 40×109/L except for 1 case in which the platelet count was reduced to 8×109/L. The median percentage of CD34+ cells in one period of collection was 0.85(0.34-1.2)%. The median MNC was (3.68±1.56)×1010. The cumulative total of mononuclear cells was (6.01±2.26)×108/kg, and the median CD34+ cell count was 4.44(2.7-7.10)×106/kg. There were no statistical differences between the 2 groups in the median percentage of CD34+ cells, the median MNC, the cumulative total of mononuclear cells and the median CD34+ cell counts (P>0.05). The average acquired time for granulocyte engraftment was 10.00(9.00-11.00) days, and for platelet engraftment was 12.50(11.00-17.25) days, with no statistical difference(P>0.05). No death occurred during the process of transplantation. DA-EPOCH therapy combined with G-CSF can effectively mobilize the peripheral blood hematopoietic stem cells in NHL patients with higher safety and lower price, and proves to be worth recommending in clinical use.

Myocardial infarction does not affect circulating haematopoietic stem and progenitor cell self-renewal ability in a rat model.

What is the central question of this study? Although peripheral blood haematopoietic stem and progenitor cells are potentially important in regeneration after acute myocardial infarction, their self-renewal ability in the post-acute phase has not yet been addressed. What is the main finding and its importance? In rat peripheral blood, we show that myocardial infarction does not negatively affect circulating haematopoietic stem and progenitor cell self-renewal ability 2 weeks after acute infarction, which suggests a constant regenerative potential in the myocardial infarction post-acute phase. Given the importance of peripheral blood haematopoietic stem and progenitor cells (HPCs) in post-acute regeneration after acute myocardial infarction (MI), the aim of the present study was to investigate the number and secondary replating capacity/self-renewal ability of HPCs in peripheral blood before and 2weeks after MI. In female Lewis inbred rats (n=9), MI was induced by ligation of the left coronary artery, and another nine underwent sham surgery, without ligation, for control purposes. Myocardial infarction was confirmed by troponinI concentrations 24h after surgery. Peripheral blood was withdrawn and fractional shortening and ejection fraction of the left ventricle were assessed before (day0) and 14days after MI or sham surgery (day14). After mononuclear cell isolation, primary and secondary functional colony-forming unit granulocyte-macrophage (CFU-GM) assays were performed in order to detect the kinetics of functional HPC colony counts and cell self-renewal ability in vitro. The CFU-GM counts and cell self-renewal ability remained unchanged (P>0.05) in both groups at day14, without interaction between groups. In the intervention group, higher day0 CFU-GM counts showed a relationship to lower fractional shortening on day14 (ρ=-0.82; P<0.01). Myocardial infarction did not negatively affect circulating HPC self-renewal ability, which suggests a constant regenerative potential in the post-acute phase. A relationship of cardiac contractile function 14days after MI with circulating CFU-GM counts on day0 might imply functional colony count as a predictive factor for outcome after infarction.

Critères de non-qualification des donneurs des cellules souches hématopoïétiques : recommandations de la Société francophone de greffe de moelle et de thérapie cellulaire (SFGM-TC)

The evolution of HLA typing and transplantation techniques makes it easier to identify a donor for hematopoietic stem cell (HSC) transplantation. This activity, strongly regulated by regulatory or normative texts, implies in addition biological, medical, para-medical and sometimes psychological evaluations. The benefit/risk discussion is complicated because it must take into account the benefit/risk ratio for the recipient, and the donor risk. No Evidence-Based Medicine data is available and serious events are very rare situations. Biovigilance declarations and their analysis are of fundamental importance. Certain obvious and definite contraindications could be detected very early in the process. It is important to assess whether a risk factor or pathology contributes to increasing the risk associated with collection. In case of recipient risk, the situation should be discussed with the patient team. These recommendations focus on adult peripheral blood HSC donors. They refer to donor information, confidentiality of exchanges, the impact of moral or material pressures, declarations of biovigilance, collegiality and traceability of difficult decisions, desirable experience and training for doctors in charge, use of expert advice informed by an explicit exchange on the possible risks, parsimony of therapeutic interventions and minimization of risks for the donor. We also recommend creation, availability and use by the community of tools and documents (registries, questionnaires, synthetic recommendations, feedback, and collegial qualification meetings) useful for practice.

Biological characteristics of Microvesicles Derived from Bone Marrow Mesenchymal Stem Cells and Their Capacities Supporting ex vivo Expansion of Hematopoietic Stem Cells

To explore the biological characteristics of microvesicles(MV) derived from bone marrow mesenchymal stem cells (BM-MSC) and their capability supporting ex vivo expansion of hematopoietic stem cells(HSC). The MV from cultured BM-MSC supernatant were isolated by multi-step differential velocity contrifugation; the morphological characteristics of MV were observed by electron microscopy with negative staining of samples; the protein level in MV was detected by using Micro-BCA method; the surface markers on MV were analyzed by flow cytometry. The peripheral blood HSC(PB-HSC) were isolated after culture and mobilization; the experiment was diveded into 2 group: in MV group, the 10 mg/L MV was given, while in control group, the same volume of PBS was given; the change of PB-HSC count was observed by cell counting; the change of surface markers on PB-HSC was detected dynamically by flow cytometry; the cell colony culture was used to determin the function change of PB-HSC after co-culture with MV. MSC-MVs are 20-100 nm circular vesicles under electron microscope. About 10 µg protein could be extracted from every 1×106 MSC. The flow cytometry showed that CD63 and CD44 were positive with a rate of 96.0% and 50.2%, while the HLA-DR, CD34, CD29 and CD73 etc were negative. When being co-cultured with GPBMNC for 2 days, the cell number of MV groups was 1.49±0.15 times of the control group (P>0.05). When being co-cultured for 4 days, the cell number of MV groups was 2.20±0.24 times of the control group(P<0.05). The CD34+ cell number of MV groups was 1.76±0.30 times the control group after culture for 2 day and 1.95±0.20 times after culture for 4 day. The MV has been successfully extracted from MSC culture supernatant by multi-step differential velocity centrifugation. MSC-MV can promote HSC expansion in vitro.

Study on the expression of TRIP13 mRNA in chronic lymphocytic leukemia B lymphocyte and the molecular mechanism of TRIP13 mediated JVM-2 cell proliferation and apoptosis

目的探讨甲状腺激素受体相互作用分子13（TRIP13）基因表达和慢性淋巴细胞白血病（CLL）的相关性，验证TRIP13基因在CLL发生发展中发挥的生物学功能，探讨TRIP13基因调控CLL发生发展的下游分子机制。方法①应用实时荧光定量PCR检测30例CLL患者和12名造血干细胞供者（正常对照组）外周血CD19+ B淋巴细胞TRIP13 mRNA的表达水平。②使用慢病毒介导的shRNA干扰JVM-2细胞TRIP13 mRNA和蛋白表达水平，同时使用随机序列shRNA处理JVM-2细胞作为对照组，分别应用MTT法和流式细胞术检测TRIP13干扰组、对照组JVM-2细胞的增殖和凋亡情况。③Western blot法检测TRIP13干扰对JVM-2细胞凋亡相关蛋白表达的影响。结果①30例CLL患者外周血B淋巴细胞TRIP13 mRNA表达水平（2−ΔCt=0.014 89）高于正常对照组（2−ΔCt=0.000 19）（P<0.001）。②在JVM2细胞中筛选出有效干扰shRNA靶点，下调TRIP13的表达能够有效抑制JVM-2细胞增殖并促进细胞凋亡。③干扰TRIP13后JVM-2细胞Myc和Bcl-2蛋白表达下调（P<0.001），Bax、caspase3、Bad蛋白表达上调（P<0.001）。结论CLL患者外周血CD19+ B淋巴细胞TRIP13 mRNA表达上调；TRIP13基因通过影响细胞增殖和凋亡相关蛋白表达而调控JVM2细胞的增殖与凋亡。

Biological Characteristics of Microvesicles Secreted by Human Peripheral Blood Hematopoietic Stem Cells

To investigate the effects of microvesicles(MV) isolated from human peripheral blood hematopoietic stem cells(PB-HSC) on immune regulation and hematopoiesis. PB-HSCs were separated by density-gradient centrifugation and cultrued. The supernatants of PB-HSC at 48 h were harvested for isolation and purification of MV by using ultracentrifugation. The electron microscopy was used to observe the morphology of MV. The protein level in MV was quantified through bicinchoninic acid(BCA) protein assay. Flow cytometry was used to detect the immunophenotype of MV. Human peripheral blood mononuclear cells(PB-MNC) were isolated from healthy donor and treated with isolated MV. After being co-cultured for 12 h, confocal microscopy was used to observe the action mode of MV on PB-MNC. After being co-cultured for 48 h, the levels of IL-2, IL-6, IL-8, IL-10, IFN-γ and TNF-α were detected by ELISA. Flow cytometry was used to detect the changes of T cell subsets and the activation of T cell subsets as well as intracellular cytokine staining after co-culture for 48 h. The methylcellulose was used to assess the hematopoiesis-supportive function of MV as well as co-cultured supernatants. The eletron microscopy revealed that MV were elliptical membrane vesicles. The protein amount in MV ranges from 29 to 110 µg. Flow cytometry showed that MV expressed mix markers on the surface, especially highly expressed MV specific marker CD63(85.86%) and hematopoietic stem cell marker CD34(33.52%). After being co-cultured for 12 h, confocal microscopy showed that MV were merged with PB-MNC. After being co-cultured for 48 h, ELISA showed that the secretion of cytokines IL-6,IL-8, IL-10 as well as TNF-α was increased while the level of IL-2 and IFN-γ was not changed much. The results of flow cytometry showed that there was no significant change in T cell subsets and T cell activation. Staining of intracellular factor showed that IL-8 was increased significantly in CD11c+ cells. The colony-forming experiments revealed that MV and the co-cultured supernatants could facilitate the colony formation. MV isolated from PB-HSC have immune-regulatery function and can prornote hematopoiesis.

G-CSF inhibits LFA-1-mediated CD4+ T cell functions by inhibiting Lck and ZAP-70.

In this study, we showed that G-CSF mobilization increased the frequency of T cells, specifically CD3+CD4+ T cells. G-CSF mobilization decreased the secretion of inflammatory cytokines of CD4+ T cells through the LFA-1/ICAM-1 signaling pathway, whereas it did not alter the TH1/TH2 ratio. We found that G-CSF mobilization inhibited LFA-1-mediated CD4+ T cell polarization and motility. In vitro, G-CSF stimulation also attenuated the polarization and adhesiveness of CD4+ T cells through the LFA-1/ICAM-1 interaction. Further investigation revealed that G-CSF mobilization suppressed LFA-1 signaling by down-regulating Lck and ZAP-70 expression in CD4+ T cells, similar results was also confirmed by in-vitro studies. These findings suggested that G-CSF directly suppressed LFA-1-mediated CD4+ T cell functions through the down-regulation of Lck and ZAP-70. The immunosuppressive effect of G-CSF mobilization deepened our understanding about peripheral blood hematopoietic stem cell transplantation. LFA-1/ICMA-1 pathway may become a potential target for graft-versus-host disease prophylaxis.

Comparison of Amicus and COBE Spectra for allogenic peripheral blood stem cell harvest: Study from tertiary care centre in India

IntroductionMost common source of stem cell graft for both autologous and allogenic haematopoietic transplants are peripheral blood haematopoietic progenitor stem cells. Adequate collection of the CD34+ cells and safety of the allogenic donor during the leukapheresis are of prime importance to an apheresis physician. Our retrospective analysis is a comparison between of two platforms namely, COBE Spectra and Amicus, for CD34+ mononuclear cell collection. Material and methodThe study included the data of GSCF (Granulocyte-Colony-Stimulating Factor) mobilized allogenic PBSC collections at our centre from January 2015 to June 2016. The apheresis platforms used were COBE Spectra and Amicus. Blood cell counts were done using LH750 Beckman Coulter (Florida, Miami, USA). CD45+ & CD34+ cell counts were done using BD FACS Canto-II Flow-Cytometer by ISHAGE guidelines. ResultsA total of 170 PBSC (100 COBE Spectra & 70 Amicus) harvests were done on 143 donors, of which 116 completed the collection in a single session and 27 required a second session. Demographic details and pre harvest peripheral blood counts for both the groups did not show any statistical differences. Amicus processed higher blood volume with higher ACD exposure and procedure time compared to COBE Spectra. Higher platelets loss was with COBE Spectra harvests with higher product volumes collection. Collection efficiency (CE2), collection ratio, CD34+ cells dose was similar on both the platforms. RBC contamination, absolute lymphocyte and monocytes counts were significantly higher with Amicus harvest product compared with COBE Spectra. A total of 14 (8.2%; citrate toxicity) adverse reactions were reported out of 170 allogenic PBSC collections. Discussion/conclusionOur study suggests that both Amicus and COBE Spectra platforms offer comparable results for allogenic PBSC collections. Amicus offers a concentrated PBSC product with lesser volume and platelets loss but higher RBC contamination.

Salvage Trerapy for Patients with Relapsed and Refractory Lymphoma by Allogeneic Hematopoietic Stem Cell Transplantation

To assess the safety and efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in treating patients with relapsed and refractory lymphoma. Thirty-one consecutive patients with relapsed or refractory lymphoma received allo-HSCT. Used conditioning regimens included conditioning based on BEAM regimen(12 cases), conditioning based on modified Bu/Cy regimen(11 cases), conditioning based on Cy/TBI regemen(6 cases) and conditioning of Bu/Cy regimen(1 case). For provention of GVHD, the MMF was used on the basis of classcal protocol consisting of CsA combined with MTX. The infused HSC included the HLA-matched related HSC(11 cases), HLA nonidentical related HSC(13 cases) and HLA-matched unrelated HSC(6 cases). The bone marrow plus peripheral blood HSC were infused in 21 cases, while only peripheral blood HSC were infused in 9 cases. Among the 31 cases of relapse/refractory lymphoma, 18 patients were male and 13 were female, 4 cases were Hodgkin's lymphoma and 27 cases were non-Hodgkin's lymphoma. ALL of the 31 patients were qualified, as they were not in complete remission (CR) or in advanced stage at the time of transplantation. Twenty-seven evaluable patients showed the engraftment of both neutrophil and platelet at a median of 12 days(range 10-20) and 13 days(range 9-34) respectively, 9 cases developed II-IV aGVHD, and cGVHD was observed in 3 patients, 5 patients can not achieve CR at 3 months after transplantation, and 6 patients relapsed after CR, the median follow-up of all the 31 patients after transplantation was 11.5 months (ranged, 0-141 months), and the 2-year OS was 46.1%±9.5% with median survival of 40 (9-141) months in the 15 survivors. The age (P<0.05), disease status before transplantation (P=0.020) and remission after transplantation(P=0.000) were significantly related with survival. Cox's proportional hazards regression model analysis showed that the age (P=0.041) and disease statue (P=0.020) before allo-HSCT were independent predictive factors for survival. Allo-HSCT is an optimal treatment strategy for the patients with relapsed and refractory lymphoma who failed to most, if not all, available options.

65 - Infusion of Autograft Natural Killer Cells/CD14+hla-DRDIM Myeloid-Derived Suppressor Cells Ratio Predicts Survival in Non-Hodgkin Lymphoma Undergoing Autologous Peripheral Blood Hematopoietic Stem Cell Transplantation (APHSCT)

65 - Infusion of Autograft Natural Killer Cells/CD14+hla-DRDIM Myeloid-Derived Suppressor Cells Ratio Predicts Survival in Non-Hodgkin Lymphoma Undergoing Autologous Peripheral Blood Hematopoietic Stem Cell Transplantation (APHSCT)