bluuebunny/crossref_metadata_2025_split huggingface dataset:
Histone Acetylation at the β-Globin Locus Changes with Developmental Age.
Authors: Barkess Grainne , Sun Jin , Fang Xiangdong , Cao Hua , Li Qiliang , Stamatoyannopoulos George 
Published: April 2020

Abstract
The β-globin locus contains a number of developmentally regulated genes that undergo switching of expression. In the embryo the ε-globin gene is expressed with a subsequent switch to γ-globin in the fetal liver, followed by a final switch to β-globin in definitive cells. Recently, the importance of chromatin structure in regulating gene expression has been recognized, and it has been proposed that modifications of histones (i.e. acetylation, methylation) have a direct effect on gene regulation. This hypothesis would suggest that changes in the histone code underlie the differences in globin gene expression during development. To address this we studied the pattern of histone acetylation across the β-globin locus at different stages of human development. Two different time points of human fetal erythropoiesis (approximately 55dpc and 98dpc) were studied. At these stages the level of γ mRNA is approximately 98%, there is no detectable ε mRNA, while β mRNA is about 2% of total mRNA. The adult stage of erythropoiesis was represented by erythroblasts derived from cultured adult CD34+ cells where γ mRNA was approx. 25% of total mRNA. Nuclei from the fetal and adult erythroblasts were analyzed by Chromatin Immunoprecipitation (ChiP) using antibodies to histone H3 acetylation and H4 acetylation. Quantification of enrichment was carried out by SYBR green real-time PCR, with PCR primers spanning the globin locus. The LCR region showed high levels of enrichment for acetylation in all stages of development. The ε-globin gene showed minimal levels of enrichment in all stages of development. In the 55dpc human erythroblasts γ-globin showed the highest level of enrichment of all globin genes, with the acetylation spreading across the promoter as well as the whole γ-coding region. In the 98dpc fetal erythroblasts, the acetylation at γ-globin was also enriched.
In the cultured adult erythroblasts a reduced level of γ-globin acetylation was present, which was expected since there was less γ-globin synthesis in these samples. There was a higher level of acetylation at the δ- and β-globin genes in the adult cells, as would be expected for the adult stage of development. These results demonstrate that dynamic changes in histone acetylation take place as the human β-globin gene region undergoes its developmental switches. Whether these changes represent primary events in the course of globin gene switching remains to be determined.
Single Administration of Antiapoptotic Cytokines Soon after High Dose Irradiation Prevents Monkeys from Thrombocytopenia.
Authors: Drouet Michel , Grenier Nancy , Delaunay Christophe , Mourcin Frederic , Mayol Jean-Francois , Lataillade Jean-Jacques , Herodin Francis 
Published: April 2020

Abstract
Preservation of hematopoietic stem and progenitor cell survival is required for recovery from radiation-induced myelosuppression. We recently showed the capacity of a single injection of antiapoptotic cytokines, namely stem cell factor, FLT-3 ligand, megakaryocyte growth and development factor, and interleukin-3 in combination (4F, each factor given intravenously at 50 μg/kg) administered shortly after a 5 Gy gamma total body irradiation to prevent monkeys from myelosuppression. As cytokine efficacy depends on the residual stem cell pool, the aim of the present study was to assess the efficacy of 4F in case of high dose irradiation. Adult monkeys (n=7) were globally and frontally irradiated at 7 Gy gamma using a Co60 source (dose rate 20 cGy/mn). Three animals received 4F 2 hours after irradiation and four control animals were injected with the diluant at the same time. Moreover, one of the treated monkeys was grafted with allogeneic mesenchymal stem cells (MSC) to enhance hematopoietic recovery. Human MSCs were infused directly in humerus simultaneously with 4F injection, allowing cell tracking. Treated monkeys experienced a very short period of thrombocytopenia as compared with untreated animals (platelet [PLT] count &lt; 20 x 109/L: 1 ± 1.7 day versus 27 ± 18 days). Areas under the curve (AUC) of PLTs and red blood cells (RBCs) between days 0 and days 90 were higher in treated animals than in controls. The animal treated with 4F + MSC did not differ from the two 4F monkeys. In contrast with 5 Gy irradiated animals previously studied, both treated and untreated monkeys experienced a prolonged period of neutropenia (neutrophil [ANC] count &lt; 0.5 x 109/L: 12 ± 3 days versus 10.3 ± 7.3) and AUCs of ANC did not significantly differ. This study shows that the 4F treatment, as a single dose regimen, prevents thrombocytopenia in high dose irradiation setting and could act as an emergency treatment for nuclear accident or terrorism victims.
Pretreatment Serum Soluble Interleukin-2 Receptor Levels Predict the Response after Immunosuppressive Therapy of Aplastic Anemia.
Authors: Min Chang-Ki , Lee Jong-Wook , Min Woo Sung , Kim Chun Choo , Kim Myungshin , Lim Jihyang , Kim Yonggoo , Han Kyungja 
Published: April 2020

Abstract
Severe aplastic anemia (SAA), characterized by pancytopenia and a hypocellular marrow, has heterogenous pathophysiologic mechanisms including a stem-cell defect or a T-cell disease. Immunosuppressive (IS) therapy based on antilymphocyte globulin (ALG) and cyclosporine (CsA) improved the response rate in patients with SAA not eligible for stem cell transplantation. The goal of IS regimens in SAA is elimination of T cells effecting the immune-mediated destruction of hematopoietic progenitor and stem cells. Increased levels of soluble interleukin-2 receptor (sIL2R) have been detected in patients with high immune activity such as autoimmune diseases, allograft rejection, and severe infection. Twenty-eight patients with SAA (15 males and 13 females, median age 27 years, range 16–62) were given an ALG (Lymphoglobuline, Institut Pasteur Merieux, 15mg/kg/day intravenously from day 1 to 5) and CsA (5mg/kg/day orally from day 12 to 180). We measured the serum sIL2R levels using ELISA test kit (CELLFREE IL-2R Medex, Kyowa, Japan) in 28 patients with SAA before and after therapy with ALG+CsA. The mean serum sIL2R levels at baseline, 7 and 60 days after ALG treatment were 572.8 pg/ml (range 181–1715), 2156.3 pg/ml (range 211–7315) and 488.0 pg/ml (range 168–1411), respectively. The baseline sIL2R levels were significantly higher in our patients when compared with normal control subjects (n=26, 379±164 pg/ml, P=0.01). Overall response, defined as transfusion independence, was achieved in 12/28 (42.9%) patients on 180 days after treatment. The baseline serum sIL2R levels were elevated more than the normal control mean+1SE (541 pg/ml) in 7 of 12 patients with response but in 4 of 16 without response (Pearson Chi-Square, P=0.047). According to the baseline sIL2R levels, the positive predictive value (the number of patients with response/&gt;control mean+1SE) was 63.6% (7/11) and the negative predictive value (the number of patients who failed to obtain the response/&lt;control mean+1SE) was 70.6% (12/17). The sIL2R levels on both day 7 and 60 did not show any difference between the groups. Overall, these findings demonstrate that serum sIL2R levels increase in patients with aplastic anemia. Pretreatment serum levels of sIL2R might be closely related to clinical outcome following IS therapy, reflecting the immunological abnormalities in individual patients.
CERA (Continuous Erythropoietin Receptor Activator): Dose-Response Trial of Subcutaneous (SC) Administration Once Every 3 Weeks (Q3W) to Patients with Aggressive Non-Hodgkin’s Lymphoma and Anemia Receiving Chemotherapy.
Authors: Osterborg Anders , Hellmann Andrzej , Couban Stephen 
Published: April 2020

Abstract
CERA is an erythropoiesis-stimulating agent (ESA) acting differently at the receptor level with a prolonged half-life. In this ongoing, multicenter, randomized, open-label, Phase II dose-response study, CERA was administered subcutaneously in a Q3W schedule to 93 transfusion-independent patients with aggressive (intermediate or high grade) B-cell non-Hodgkin’s lymphoma (NHL) and anemia receiving combination chemotherapy. Eligible patients met the following inclusion criteria: age ≥18 years, hemoglobin (Hb) &lt;11 g/dL, combination chemotherapy scheduled to be administered throughout the 12-week treatment period, life expectancy &gt;6 months, and Eastern Cooperative Oncology Group (ECOG) performance status grade 0–2. Major exclusion criteria included transferrin saturation &lt;20% and platelet count &lt;50 x 109/L. No patient had received an ESA in the 8 weeks prior to the first dose of CERA. Patients were randomized to receive CERA 2.1 μg/kg (n=31), 4.2 μg/kg (n=30), or 6.3 μg/kg (n=32) administered once every 3 weeks for 12 weeks. The primary efficacy variable was time-adjusted average change in Hb from baseline during 12 weeks, until end of initial treatment (last observed value before dose change or transfusion). Enrollment has been completed; the treatment phase of the trial is nearing conclusion. Demographics show similar baseline characteristics in all three patient subgroups [mean age: 62.9 years (2.1 μg/kg); 59.1 years (4.2 μg/kg); 64.3 years (6.3 μg/kg)]. Diffuse large B-cell lymphoma was the most common lymphoma in all three subgroups (87%, 70%, and 84% of patients in the 2.1, 4.2, and 6.3 μg/kg subgroups, respectively). To date, the majority of patients receive an anthracycline-containing chemotherapy regimen, either standard CHOP or CHOP plus rituximab. Preliminary efficacy and safety results will be presented. Ongoing assessments have indicated a safety profile consistent with that seen in patients with aggressive NHL and anemia receiving chemotherapy. This Phase II trial will help further characterize the ability of CERA to safely correct anemia when administered at an extended dosing interval (Q3W) to patients with aggressive NHL receiving chemotherapy.
Inhibition of T Lymphocyte (TL) Proliferation by Mesenchymal Stem Cells (MSCs): Role of Cell Contact between MSCs and TL, Role of MSCs Dose Effect, and Role of Soluble Factors and Adhesion Molecules Expressed by MSCs.
Authors: Nasef Aisha , Chapel Alain , Benshidom Morad , Zang Yizhuo , Mazurier Christelle , Bouchet Sandrine , Lopez Manuel , Gorin Norbert Claude , Thierry Dominique , Fouillard Loic 
Published: April 2020

Abstract
Mesenchymal stem cells (MSCs) can inhibit T lymphocyte (TL) proliferation in mixed lymphocyte reaction (MLR) and are candidate to induce tolerance of allogeneic haematopoietic stem cell graft. Results of previous studies evaluating the role of cell contact between MSCs and TL are contradictory. Some soluble factors produced by MSCs have been shown to be involved in TL inhibition such as Transforming Growth Factor-b (TGF-b), Hepatocyte Growth Factor (HGF), and Indoleamine 2,3 dioxygenase (IDO).
In this study inhibition of TL proliferation in MLR was evaluated in cultures with MSCs-TL contact and in cultures without MSCs-TL contact (transwell), at different concentration of MSCs (ratio MSCs/TL: 0.3, 0.1, 0.03, 0.01). Expression of mRNA encoding cytokines and adhesion molecules produced by MSCs were analysed by semi-quantitative RT-PCR. Inhibitory cytokines studied were IDO, HLA-G, LIF, IL-10, TGF-b and HGF; adhesion molecules studied were VCAM and LFA3.
Results of MSCs dose effect. In MLR with MSCs-TL contact, inhibition of proliferative index was related to the dose of MSCs. The percentage of inhibition was 74%, 60%, 48%, and 28% at ratio 0.3, 0.1, 0.03 and 0.01 respectively (p&lt;0.05). In MLR without MSCs-TL contact, no dose effect was observed: the percentage of inhibition was 48%, 46% and 46% at ratio 0.3, 0.1 and 0.03 respectively; no inhibition was observed at ratio 0.01. Results of comparison between inhibition with cell contact and without cell contact. At ratio 0.3 the percentage of inhibition was 74% with cell contact and 48% without cell contact (p&lt;0.05), at ration 0.1 the percentage of inhibition was 60% and 46% respectively (p&lt;0.05). These results confirmed that TL inhibition is mediated by soluble factors but is increased when MSCs and TL are in contact. Results of mRNA RT-PCR in MSCs showed an overexpression of IDO, HLA-G and LIF in cultures with cell contact and without cell contact. In cultures with cell contact an overexpression of IL10 and TGF-b was observed, but not in cultures without cell contact. Adhesion molecules VCAM and LFA3 were overexpressed in both types of cultures with and without cell contact. In conclusion although cell contact is not mandatory to inhibit T cell proliferation, inhibition is higher when TL and MSCs are in contact, concomitantly an overexpression of additional inhibitory molecules is observed.
Stimulation of the Supportive Function of Marrow Stromal Cells by Anticancer Agents: Critical Role of Marrow Macrophages.
Authors: Ohta Kensuke , Nakamae Hirohisa , Koh Ki-Ryang , Nakaie Kiyotaka , Li Hong-Zhang , Yamane Takahisa , Takubo Takayuki , Hino Masayuki 
Published: April 2020

Abstract
Anticancer agents used for the treatment of hematological malignancies are known to cause long-term toxicities to the structure and function of bone marrow microenvironment. These obbservations are based predominantly on the in vivo experiments with anticancer drug-treated mice and also on the clinical features of the patients who survived after inhtensive chemotherapies. In the present study, in order to evaluate immediate gene response of human marrow stromal cells to anticancer drugs, we tested with real-time PCR for the expression of various cytokine mRNAs from the stromal cells treated with anticancer drugs. Contrary to our expectation, we found that treatment of stromal cells with cytarabine (Ara-C; 0.1–100 μmol) for 3 to 14 days significantly and dose-dependently enhances the expression of mRNAs of stem cell factor (SCF) and leukemia inhibitory factor (LIF) while downregulates interleukin-6 (IL-6) mRNA. On the other hand, carboplatin (0.1–100 μmol) up-regulated only SCF mRNA and adriamycin (0.001–1 μmol) did not affect these gene expressions (n=6). In order to determine the responsive cell elements of stromal cells for these novel responses to Ara-C, mRNAs were independently quantified after separating CD14 positive macrophages and CD45 negative mesenchymal cells from Ara-C-treated stromal cells (n=6). In this additional experiments, we found that the above-mentioned changes in gene expression by Ara-C were provoked predominantly in the stromal macrophage fraction. Based on these observations, we treated the stromal cells established from 6 patients with AML and 11 with non-leukemic subjects with Ara-C for 2 weeks, followed by washing 3-times to eliminate Ara-C. Allogenic CD34 positive cells were then recharged and supportive functions of the stromal cells were evaluated with standard 2-stage long-term cultures. Indeed, the results showed that transient treatment with Ara-C significantly and dose-dependently upregulates supporting function of premature hematopoietic cells in non-leukemic and, although to the less extent, in leukemic stromal cells. These novel short-term stimulatory effects of Ara-C to marrow microenvironment may provide new explanations for some clinical events such as mechanism of rapid recovery of normal hematopoiesis and stem cell mobilization observed several days after completion of intensive chemotherapy to acute leukemia. In addfition, the pattern of gene response of stromal cells induced by Ara-C is of biologically great interest since it is quite different from the already known gene response of human stromal cells provoked by IL-1.
Immortalized Human Placenta Derived Mesenchymal Cells by Transfection with Bmi-1 Lost the Abilitiy To Differencitate into Cell Types of Osteocytic, Condoricytic and Neural Lineages.
Authors: Zhang Xiaohong , Soda Yasushi , Takahashi Kenji , Bai Yuansong , Igura Koichi , Mitsuru Ayako , Tojo Arinobu , Takahashi Tsuneo A. 
Published: April 2020

Abstract
Introduction: We previously reported that mesenchymal cells derived from chorionic villi of human placenta could differentiate into various cell types including osteocytic, condoricytic and neural lineages. However, these cells can slowly proliferate with limited life span. In the present study, we transduced human placenta-derived mesenchymal cells (PDMCs) with cDNA encoding Bmi-1 and telomerase reverse transferase (hTert), respectively or simultaneously by lentiviral vector, and tested transduced cells for their ability to differentiate into multi-lineages and to support expansion of CD34+ cells in cord blood as feeder cells in the presence of cytokine cocktails.
Method: The lentivairal vector expressing either Bmi-1, hTert or hrGFP was prepared as previously. The human PDMCs were isolated from chorionic villi of human placenta using the explant culture, and were cultured in DMEM (low glucose) with 10% FBS. The established cells were infected with either of the three viruses or a combination of Bmi-1 and hTert virus at the multiplicity of infection that allowed almost 100% of infection efficiency based on hrGFP fluorescence by flow cytometry. Those infected cells and uninfected cells served as control were then continuously propagated. These cells were then transferred into the specific culture media to differentiate them to osteocyte, condorocyte and neural cells as reported previously.
Results: The control and PDMC/hTert cells showed a broad and flat shape suggesting that these cells were senescence and ceased proliferation. Cells infected with Bmi-1 and Bmi-1/hTert escaped the crisis and continued proliferation. These cells have been maintained in culture for more than 1 year and continued to proliferate near to 100PD. We found that Bmi-1 over-expression could induce endogenous telomerase activity and elongate telomere length in PDMCs. The expression of p16 mRNA and protein were down-regulated in prolonged culture cells, but p14ARF was not. The differentiation potential of transduced cells was gradually lost as culture was prolonged as well as the ability to support the expansion of CB CD34+ cells. Gene expression analysis on Bmi-1- infected cells at different PD revealed that more than 100 genes were induced or repressed in several different functional categories.Extensive increase in expression of the IGFBP2 mRNA and higher concentration of IGFBP2 was observed.
Conclusion: Bmi-1 induced Telomerase activity and immorlized human placenta derived mesenchymal cells. The ability of transduced cells to differentiate into various cell types and support expansion of CB CD34+ cells were lost with propagation suggesting that subtle transformation has occurred in the Bmi-1 transduced cells.
Selective Depletion of Alloreacting CD25+ Cells from Stem Cell Allografts Can Reduce Acute Graft-Versus-Host Disease Following Matched Related Donor Transplantation.
Authors: Solomon Scott R. , Tran Thao , Carter Charles S. , Hensel Nancy , Wisch Laura , Mielke Stephan , Read Elizabeth , Ghetie Victor , Schindler John , Vitetta Ellen , Barrett John 
Published: April 2020

Abstract
Graft-versus-host disease (GVHD) remains a major cause of morbidity and mortality after allogeneic stem cell transplant (SCT), especially in older patients. We previously showed that host-reactive donor T cells are selectively depleted (SD) from an allograft ex vivo, following a short co-culture of donor cells with irradiated T cell stimulators from the recipient and subsequent treatment with an anti-CD25 immunotoxin. We report a pilot study to test the hypothesis that GVHD could be decreased in a cohort of elderly patients receiving SD allografts from HLA-identical sibling donors. Sixteen patients, median age 65 years (range 51–73), with advanced hematologic malignancies were transplanted following reduced-intensity conditioning with fludarabine and either cyclophosphamide (n=5), melphalan (n=5), or busulfan (n=6). Cyclosporine was used as the only additional GVHD prophylaxis. SD allografts contained a median CD34 dose of 4.5x106/kg (range 3.5–7.3) and an SD CD3 dose of 1.0x108/kg (range 0.2–1.5). Fifteen patients achieved sustained engraftment. The helper T lymphocyte precursor (HTLp) frequency assay demonstrated depletion of host-reactive donor T cells in 9/11 cases tested from a mean of 1/182,089 to 1/822,354 (mean 5.5-fold depletion), while third party responses were conserved. Kaplan-Meier estimates of probability of grade II-IV and grade III-IV acute GVHD were lower than those seen in a historical control group of patients receiving cyclosporine alone for GVHD prophylaxis (35±13% vs. 57±10%, p=0.34) and (7±6% vs. 38±6%, p=0.05), respectively. Of note, the two patients who developed visceral (gut ± liver) GVHD showed ineffective allodepletion by HTLp (figure). Chronic GVHD occurred in five of 14 evaluable patients. At a median follow-up of 212 days (range 60 – 690), seven of sixteen patients remain alive and in remission. Relapse deaths occurred in four patients (refractory AML [2], therapy-related MDS [1], and CMML [1]). Non-relapse mortality in this high-risk cohort of patients included graft failure [1], GVHD [2], infection [1], and myocardial infarction [1]. In summary, CD25-directed allodepletion of stem cell allografts can reduce clinically relevant acute GVHD following matched related donor transplantation.
Figure Figure
Role of Integrins in Adhesion of Hematopoietic Progenitor Cells.
Authors: Eshghi Shawdee , Zhang Jing , Griffith Linda G. , Lodish Harvey F. 
Published: April 2020

Abstract
The hematopoietic stem cell niche is the set of soluble growth factors, cell-cell and cell-matrix interactions that contribute to stem cell self renewal in the bone marrow. While cytokines and cell-cell interactions have been well documented, cell-matrix interactions in the niche are less understood. Integrins are a class of highly conserved cell adhesion molecules that are important in hematopoietic development and homing. However the specific role of integrins in mediating adhesion to extracellular matrix in the hematopoietic stem cell niche is unknown. The terminal stages of erythropoiesis in the fetal liver provide a good model system with which to develop several of the assays to be used with HSCs. Using flow cytometry, murine fetal liver erythroid progenitors can be separated at four distinct stages of development based on expression of CD71 and Ter119. Further FACS and quantitative PCR analysis revealed that α4β1 integrin is significantly downregulated over the course of erythroid differentiation. Using a centrifugation assay, we determined that this change is accompanied by a loss of adhesion to fibronectin, and that adhesion to fibronectin is blocked by addition of anti-integrin antibodies. Finally, fetal liver progenitor cells adhered to comb co-polymer surfaces engineered to present peptides specifically recognized by α4β1 integrins. By determining the integrin profile expressed by hematopoietic stem cells and measuring stem cell adhesion to ECM in a similar manner, we can begin to understand how these specific interactions present developmental cues important to maintaining the stem cell phenotype in vivo, in addition to leading to design parameters for ex vivo culture systems.