bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hyaluronidase and IFN-γ from Activated T-Cells Are Key Mediators To Induce Immunosuppressive Activity of Mesenchymal Stem Cells.
Authors: Sato Kazuya , Katsutoshi Ozaki , Oh Iekuni , Matsu Haruko , Tatara Raine , Meguro Akiko , Hatanaka Keiko , Nagai Tadashi , Muroi Kazuo , Ozawa Keiya 
Published: October 2019

Abstract
Mesenchymal stem cells (MSC) have been shown to suppress T-cell proliferation in vitro and are being clinically applied to treatment for steroid-refractory graft-versus-host disease (GVHD). However, the molecular mechanisms of immunosuppressive activity of MSC have not been fully understood. We have recently reported that nitric oxide (NO) is one of the key molecules for T-cell suppression by MSC (Blood, 2007;228–34). Inducible NO synthase (iNOS) expression in MSC and NO production were exclusively detected upon interaction with activated T-cells. An inhibitor of NOS restored the proliferation of T-cells. We have also shown that both IFN-γ and NF-κB mediate NO production by MSC (BBRC, 2007;80–90), however the factor(s) from T-cells that induce immunosuppressive activity of MSC have not been elucidated. In the present study, we focused on hyaluronic acid (HA), a major extracellular matrix component, because
low molecular weight (LMW) HA is known to play an essential role in regulation of inflammation and infection, HA induces NOS in macrophages through a NF-κB-dependent mechanism, and T-cells are reported to express hyaluronidase-2 mRNA after activation.
A series of such information suggests that activated T-cells regulate immunosuppressive activity of MSC through HA signaling. We investigated the molecular events in the interaction between MSC and activated T-cells, and demonstrated that HA may play an important role in the scenario. First, ELISA showed that MSC produce a large amount of HA. RT-PCR revealed that among three hyaluronic acid synthase (HAS), mRNA for HAS-1 but neither HAS-2 nor HAS-3 is constitutively expressed by MSC. Flow cytometric analysis also showed that MSC express CD44, one of the receptors for HA. In macrophages, either IFN-γ or LMW HA is known to induce NO production. Therefore, we investigated whether IFN-γ or LMW HA can induce NO production via NF-κB signaling in MSC. As a result, only in the presence of IFN-γ, LMW HA but not high molecular weight (HMW) HA induced expression of iNOS in MSC, followed by NO production from MSC. We found that prostaglandin E2 (PGE2), which was previously reported as another immunosuppressive factor of MSC, is also induced by the combination of IFN-γ and LMW HA. Since hyaluronidase activity in activated T-cells has already been reported, these data support a novel hypothesis that HMW HA produced by MSC is digested by hyaluronidase from activated T-cells, and that resultant LMW HA cooperates with IFN-γ from activated T-cells in inducing iNOS expression in MSC. Locally produced NO through the above mechanisms may suppress activated T-cell proliferation. These findings would be important to further potentiate and regulate the immunomodulatory activity of MSC in a variety of applications.
Human B Cell Development from Adult Bone Marrow Is Profoundly Dependent on IL-7 Due to Decreased B Lymphoid Generative Capacity.
Authors: Parrish Yasmin K. , Baez Ineavely , Milford Terry-Ann M. , Rogerio Jaqueline W. , Sahakian Eva , Kagoda Mercy , Benitez Abigail , Huang Grace , Payne Kimberly J. 
Published: October 2019

Abstract
IL-7 is critical for normal murine B cell development, however its role in human B lymphopoiesis is controversial. Here we examine the role of IL-7 in human B cell development using a novel in vitro model based on co-culturing cord blood (CB) and adult bone marrow (BM) hematopoietic stem cells (HSCs) on primary stroma from adult human BM. Addition of IL-7 to this co-culture model increased B cell production by ∼40-fold. IL-7-induced increases were dose-dependent and specific to CD19+ cells. Flow cytometry analysis of STAT5 phosphorylation, and expression of the IL-7Rα and the proliferation antigen, ki-67, indicate that IL-7 acts directly on B cell precursors and that this effect is at least partially mediated through increased proliferation. Ongoing studies are assessing the effects of IL-7 on apoptosis among B cell precursors. IL-7 effects appeared to be most profound in cultures initiated with adult human BM HSCs where few, if any, human B lineage cells are generated in the absence of either mouse or human IL-7 activity. However, when co-cultures initiated with CB and BM HSCs were compared, IL-7-induced increases were similar in magnitude, and B cell precursors responded similarly to IL-7. Therefore we compared CB and BM HSCs with respect to B lymphoid generative capacity. The ability of CB and BM HSCs to generate CD19- progeny in our cultures was not significantly different, however the in vitro B lymphoid generative capacity of adult BM HSCs was 40 times less than that of CB HSCs. Our results provide evidence that IL-7 is critical for B cell production from HSCs in adult BM due to their low B lymphoid generative capacity as compared to HSCs in CB. These studies have implications for B cell recovery following transplantation with stem cells from CB versus BM and suggest that B cell recovery could be impacted by availability of IL-7.
In Patients with Aplastic Anaemia, Conditioning with Reduced Intensity Followed by Infusion of T-Cell Depleted Grafts Is Associated with Limited Transplant Related Morbidity and Universal Survival.
Authors: Novitzky Nicolas , Thomas Valda , Du Toit Cecile E. , McDonald Andrew B. 
Published: October 2019

Abstract
Introduction: While the survival of patients receiving allogeneic stem cell transplantation for aplastic anaemia has improved significantly, a substantial number of patients develop extensive graft vs. host disease (GvHD), with the associated morbidity and mortality. T-cell depletion of grafts decreases the incidence and severity of GvHD but may result in greater risk of graft failure.
Patients and methods: Between 2004 and 2007, seventeen patients with marrow aplasia were conditioned with fludarabine 30 mg/m2 for 5 consecutive days (150 mg/m2) followed by cyclophosphamide 120 mg/kg. In addition 2 (one with Fanconi’s anaemia) received busulfan 8 mg/kg. All had received more than 30 units of blood products and 2 had lymphocytotoxic antibodies that required HLA matched platelet transfusions. HLA identical sibling donors were mobilised with filgrastim (5–10 ug/kg) for 5 days and underwent large volume apheresis. Donor cells were quantitated and the CD34+ cell number determined. PBPC grafts were then incubated “in the bag” with alumtuzumab (1 mg/1010 mononuclear cells) at 20°C for 30 minutes and infused without further processing. Patients commenced cyclosporin from day −1 and continued until day 90 post transplantation. End points were engraftment, OS, DFS, incidence and severity of GvHD.
Results: Median age was 22 (7–49) years and 5/17 were females. Median CD34+ cell infused was 3.93 x 106/kg, containing median of 22.2 x 14/kg CFU-GM. The median “ex vivo” alumtuzumab dose employed was 10 mg (range 5–10 mg). There was universal engraftment and leukocytes reached 0.5 x 109/L at a median of 14 days (8–17) post transplantation. Other than haematological suppression, the toxicity of the conditioning was limited to grade I–II intestinal side effects. Seven individuals did not develop pyrexia following infusion of the graft. No patient developed GvHD. Two subjects had CMV reactivation (pp65+) and responded to therapy with gancyclovir. Another two developed haemorrhagic cystitis due to BK virus. Donor-recipient sex mismatch was present in 8 instances and post transplant bone marrow karyotype analyisis showed that haematopoiesis was of donor’s origin in each case. One patient developed delayed graft failure 6 months post transplantation and had a satisfactory recovery following a second T- cell depleted transplant with similar conditioning. Another developed mixed chimerism at 8 months and received DLI. All patients survive at a median of 630 days (range 60–1433) with normal blood parameters.
Conclusion: We conclude that purine analogues in combination with alkylators are a sufficiently intense immunosuppressive conditioning to allow successful engraftment of T-cell depleted PBPC grafts. Alumtuzumab “in the bag” led to excellent GvHD prophylaxis, leading to minimal morbidity of the procedure.
Molecular Mechanisms of Spindle Checkpoint-Apoptosis Linkage and Karyotypic Stability in Stem Cells.
Authors: Mantel Charlie , Rhorabough Sara , Guo Ying , Lee Man-Ryul , Han Myung-Kwan , Kim Kye-Seong , Broxmeyer Hal E. 
Published: October 2019

Abstract
Ex-vivo expansion of human HSC prior to bone marrow transplantation is still an unrealized goal that could greatly extend the usefulness of this mainstay strategy for treating numerous human hematologic diseases. The safety of this process for potential use in humans depends in large part on the maintenance of karyotypic stability of HSC during expansion, a lack of which could contribute to serious, even fatal, complications such as cancer, and could also contribute to engraftment failure. The spindle checkpoint and its linkage to apoptosis initiation is one of the most important cellular processes that helps maintain chromosomal stability in rapidly proliferating cell populations by removing aneuploid and karyotypically abnormal cells via activation of cell death programs. Detailed understanding of the molecular regulation of this vital cell cycle checkpoint is important to maximize safety of in-vitro HSC expansion techniques. It is widely accepted that mammalian cells enter the next G1-phase with 4N DNA after slippage from prolonged drug-induced mitotic block caused by activation of the transient spindle checkpoint that it is from this state that polyploid/aneuploid cells initiate apoptosis. However, definitive biochemical evidence for G1 is scarce or unconvincing; in part because of methods of protein extraction required for immunoblot analysis that cannot take into account the cell cycle heterogeneity of cell cultures. We used single-cell-intracellular-flow-cytometric analysis to define important factors determining cell fate after mitotic slippage. Results from human and mouse embryonic stem cells that reenter polyploid cell cycles are compared to human somatic hematopoietic cells that die after MS. We now report for the first time that phosphorylation status of pRb, p53, CDK1, and cyclin B1 levels are important for cell fate/apoptosis decision in mitotic-slippage cells, which occurs in a unique, intervening, non-G1, tetraploid subphase. Hyperphosphorylated Rb was extremely abundant in mitotic-slippage cells, a cell signaling event usually associated with early G1-S phase transition. P53 was phosphorylated at sites known to be associated with apoptosis regulation. Cyclin A and B1 were undetectable in mitotic slippage cells; yet, CDK1 was phosphorylated at sites typically associated with its activation. Evidence is also presented raising the possibility of cyclin B1-independent CDK1 activity in mitotic-slippage cells. These findings challenge the current models of spindle checkpoint-apoptosis linkages. Our new model could have important implications for methods to maintain karyotypic stability during ex-vivo HSC expansion.
Expansion of BCR-ABL Positive Leukemic Stem Cells Is Dependent on Hedgehog Pathway Activation.
Authors: Dierks Christine , Beigi Ronak , Stegert Mario , Zirlik Katja , Schmitt-Graeff Annette , Veelken Hendrik , Warmuth Markus 
Published: October 2019

Abstract
Chronic Myeloid Leukemia is characterized by the expansion of a leukemic stem cell clone carrying a Philadelphia translocation, which outgrows the non-malignant haematopoietic stem cells. Here we show that BCR-ABL directly enhances self renewal of haematopoietic stem and progenitor cells by activating the hedgehog signaling pathway through upregulation of SMO. Induction of hedgehog signaling in regular bone marrow by overexpression of SMO or loss of PTCH induces expansion of short term repopulating cells, but also enhances the expansion of long term repopulating haematopoietic stem cells. Pharmacological inhibition of SMO activity in BCR-ABL positive bone marrow cultures induced apoptosis in BCR-ABL positive differentiated cells and inhibited colony forming capacity of BCR-ABL positive self renewing cells in vitro. While loss of SMO in regular haematopoiesis only impacts the reconstitution of CD8 positive T-cells and short term repopulating cells (transplantation of SMO−/− fetal liver cells), the development of BCR-ABL positive leukemias was impaired and retransplantability abolished in the absence of SMO expression. Combined treatment of leukemic mice with AMN107 (Abl inhibitor) and Cyclopamine (SMO inhibitor) lead to a reduction of Bcr-Abl positive self-renewing cells in vivo and enhanced the time to relapse more than 3-fold compared to mice treated with AMN107 alone. Thus we conclude that BCR-ABL enhances the self renewal of leukemic stem cells through intrinsic activation of hedgehog signaling by upregulation of SMO. Therefore Hh pathway inhibition alone or in combination with Abl inhibitors could serve as an effective therapeutic strategy to reduce the malignant stem cell pool in BCR-ABL positive leukemias.
Interstitial Lung Disease in Patients with Diffuse Large B-Cell Lymphoma Receiving CHOP-Based Chemotherapy Is Associated with the Use of Rituximab.
Authors: Go Ronald S. , Riggle Kevin M. , Beier-Hanratty Sue A. , Gundrum Jacob D. , Schroeder Jonean E. , Wright Linda J. , Manske Brian A. 
Published: October 2019

Abstract
Background: Several cases of chemotherapy-induced interstitial lung disease (ILD) or pneumonitis have been reported in recent years in patients with lymphoma. The potential roles of rituximab (R) and granulocyte colony stimulating factor (GCSF), agents more commonly used in recent years, are suggested.
Objective: We wanted to determine the prevalence of ILD and identify risk factors in patients with diffuse large B-cell lymphoma (DLCL) who received cyclophosphamide/doxorubicin/vincristine/prednisone (CHOP)-based chemotherapy.
Methods: Selection criteria included newly diagnosed DLCL patients treated at our institution from 2000–2006 who received CHOP with or without R and had at least 3 serial CT or PET/CT scans during chemotherapy. ILD was defined as new bilateral interstitial pulmonary infiltrates not likely representing fluid overload or fibrosis. One radiologist blinded to clinical data reviewed all imaging studies.
Results: Out of a total of 174 new cases of DLCL, 73 met our study criteria. Reasons for exclusion included no chemotherapy (45), <3 imaging studies (46), and non-CHOP-based chemotherapy (10). Among the 73 study patients, 52 (71%) received R in addition to CHOP. Eleven (15.1%) patients developed ILD, all in the subgroup that received RCHOP (P = 0.027). Most occurred between cycles 2 and 4 of RCHOP (81.8%) and persisted until after completion of chemotherapy (63.6%). Nine (81.8%) patients with ILD were asymptomatic and never required treatment or delay of RCHOP. The remaining 2 patients became symptomatic (1 hospitalized), were empirically treated for atypical pneumonia with clinical recovery, and had delay of RCHOP. All patients received the intended number of courses of RCHOP. Univariate analysis of potential ILD risk factors among those who received RCHOP showed a trend with the subgroup that either had GCSF or cardiopulmonary disease (P = 0.09). Multivariate analysis using a two-variable model suggests that the use of GCSF or presence of cardiopulmonary disease (P = 0.065) and a high (3–5) international prognostic index score (P = 0.13) need further investigation as risk factors.
Conclusions: In our cohort of DLCL patients receiving CHOP-based chemotherapy, ILD was common and significantly associated with the use of R. While most cases were asymptomatic, self-limited, and did not require delay of chemotherapy, more serious presentation could occur. The mechanism of ILD is unknown and requires further investigation.
Expression Profile Analysis of Genes Related to Resistance/Sensibility to Prednisolone, Daunorubicin, L-Asparaginase and Vincristine in Childhood Acute Lymphoblastic Leukemia.
Authors: Silveira Vanessa S. , Scrideli Carlos A. , Moreno Daniel A. , Yunes Jose A. , Brandalise Silvia R. , Toledo Silvia , Lee Maria L.M. , Petrili Antonio S. , Odoni-Filho Vicente , Bendit Israel , Tone Luiz G. 
Published: October 2019

Abstract
Resistance to anti-leukemic agents in childhood ALL has been strongly associated with prognosis. The aim of this study was to analyze the expression profile of genes related to resistance/sensibility of 4 anti-leukemic agents and their correlations with age, diagnosis WBC count, risk group classification, molecular cytogenetics, induction treatment response and event-free survival (EFS) in ALL children. Were analyzed the expression profile of 19 genes related to resistance/sensibility to vincristine (RPRP2, CD44, TCF5, KCNN1, TIF1), prednisolo ne (F8A, CDK2AP1, BLVRB, CD69), daunorubicin (MAP3K12, SHOCK2, PDCH9, EGF1, KCNN4) and L-asparaginase (GPR56, MAN1, SCGF, IGFBP7, GATA3) by real-time quantitative PCR, using SYBR Green approach and melting curve analysis, with GUSB as housekeeping gene. Relative quantification was done using the 2-DDCT method (efficiencies from 1.9 to 2.1 to all the analyzed genes). These genes were chosen according to those described on microarray analysis by Holleman et al. (NEJM, 351:533, 2004). From 154 consecutive children admitted to ALL treatment in the 3 participating institutions, 139 had the gene expression profile analyzed. Sixteen were excluded due to lack of good quality RNA/cDNA. The follow up ranges from 36–60 months. All patients were classified and treated according the Brazilian ALL treatment protocol (GBTLI-99). Association among the variables analyzed and the gene expression levels was evaluated by Mann-Whitney and chi-square tests. EFS analysis was performed using Kaplan-Meier and log-rank test and multivariate analysis by the Cox proportional model to define independent prognostic factors. Correlation between the genes was analyzed by Spearman test. The 4-years EFS was 75,5% to all patients analyzed (81,4% to standard risk and 68,7% to high risk). Statistical correlation was observed among the genes of resistance of daunorrubicin, L-asp and vincristine (P<0,001). There is a significant association (p<0,05) between the levels of expression of the genes TIF1A, BLVRB, CD44, SCFBP1, TCFL5 and risk group; CD44, MAP3K12, SHOCK2 and CD10 antigen and GPR56, SHOCK2 with favorable event. The overexpression of SHOCK2 and GPR56 genes were also associated to better 4y-EFS in univariate (86,3% x 68,7% (p:0,04) and 86,2% x 72,2% (p:0,03) respectively) and multivariate analysis (p:0,03 and 0,09 respectively). No significant differences were observed in the other variables analyzed. The present data suggest that the expression of some analyzed genes have influence in the resistence/sensibility to the treatment and outcome in childhood ALL; especially over-expression of the SHOCK2 and GPR15 genes were associated with higher sensibility to daunorrubicin and L-asp respectively.
Evaluating the Safety of Erythropoiesis-Stimulating Agents (ESAs) in Patients with Breast Cancer, Lung Cancer, and Hematologic Malignancies: A Meta-Analysis of Randomized, Controlled Trials.
Authors: Glaspy J. , Henry D. , Vansteenkiste J. , Ludwig H. , Lillie T. , Tomita D. , Crawford J. 
Published: October 2019

Abstract
Background: Recent clinical trial data have generated safety concerns regarding ESA use, particularly as to whether the results observed in certain patients with breast cancer, lung cancer, and hematologic malignancies are generalizable to all patients with those tumor types. It is critical that the results of these studies are appropriately integrated into the total body of evidence in order to enable balanced conclusions to be drawn. We conducted a systematic literature review to assess the safety and efficacy of ESA therapy in patients with breast cancer, lung cancer, and hematologic malignancies.
Methods: The search was aligned with that conducted by the Cochrane Collaborative group (Bohlius 2006) and was therefore limited to randomized, controlled clinical trials in cancer patients that compared the use of ESAs with either observation (RBC transfusion as required) or placebo and that collected survival data (either long-term or during the study period). Results from trials were meta-analyzed using Comprehensive Meta-Analysis v 2.2 software. Heterogeneity between studies was evaluated using a chi-square test (Q). Random and fixed-effects models were used to estimate an overall odds ratio and 95% confidence interval; the inconsistency statistic, I2, is also provided.
Results: For breast cancer, seven studies were identified. Meta-analysis of data from these studies indicated an overall neutral risk of death for patients receiving ESAs, despite the large contribution (weighted at about 40% of the overall result) of the BEST study results (Leyland-Jones, 2005) (Table). Five ESA-controlled lung cancer studies were identified in CIA, one in radiotherapy, and one in anemia of cancer (AOC). In each of the studies in CIA, the ESA groups had neutral survival risks relative to the control group. Meta-analysis of the CIA, radiotherapy, and AOC lung cancer data combined indicated an overall neutral risk of death for patients receiving ESAs (Table). For hematologic malignancies, eight studies were identified (one of which had no deaths). Meta-analysis indicated an overall neutral risk of death for patients receiving ESAs (Table).
Conclusions: The results of some studies have suggested a detrimental effect on survival associated with ESA use in certain patients with breast, lung, and hematological malignancies. However, results do not appear to be generalizable to similarly designed, randomized controlled trials conducted in the same tumor types. Additionally, when these studies are integrated into the total body of evidence, ESAs have a neutral effect on survival in these patients.