bluuebunny/crossref_metadata_2025_split huggingface dataset:
The GATA-1/FOG-1/NuRD Axis Is Essential for Normal Hematopoiesis.
Authors: Hong Wei , Wang Hongxin , Wang Yuhuan , Choi John K. , Shelat Suresh G. , Poncz Mortimer , Blobel Gerd A. 
Published: April 2020

Abstract
GATA-1 controls the development of erythroid cells and megakaryocytes through its ability to activate and repress gene transcription. GATA-1 binds many nuclear proteins, but only a few of these associations have been examined in vivo. One important example is FOG-1, a critical cofactor that contributes to both gene activation and repression by GATA-1. Loss of FOG-1 generally phenocopies GATA-1 deficiency, impairing both erythroid and megakaryocytic differentiation. We reported previously that FOG-1 directly binds the NuRD protein complex, which contains histone deacetylase and chromatin remodeling activities. This provides one mechanism for GATA-1/FOG-1-mediated gene repression. Accordingly, ChIP profiling of the NuRD proteins MTA-2, RbAp46 and Mi-2β revealed the presence of these molecules at the Kit and Gata2 genes both of which are directly repressed by GATA-1 in a FOG-1-dependent manner. NuRD proteins were spread broadly across the Kit and Gata2 genes but were further enriched at sites occupied by GATA-1 and FOG-1 in vivo. Unexpectedly, we also observed NuRD components at GATA-1-activated genes including β-globin and Ahsp. Moreover, the ability of FOG-1 to augment GATA-1-induced transcription in transient transfection assays required NuRD binding. Hence, NuRD may be bi-functional, contributing to either gene activation or repression, depending on the transcriptional and cellular context. To study the role of the FOG-1/NuRD interaction in vivo we generated mice bearing missense mutations in the Fog-1(Zfpm1) gene that disrupt NuRD binding in the FOG-1 protein. Homozygous mutant mice are born at reduced Mendelian ratios. Surviving animals display ineffective erythropoiesis marked by splenomegaly and impaired erythroid maturation. In addition, homozygous mutant animals display macrothrombocytopenia with impaired platelet function. Thus, recruitment of NuRD by GATA-1 and FOG-1 is essential for both erythropoiesis and megakaryocytopoiesis. Ongoing studies include further phenotypic analysis of the mutant mice, including comparative gene expression analysis in stage-matched wild-type and mutant erythroid cells to identify critical NuRD-dependent GATA target genes, and to resolve whether NuRD is essential for both activation and repression by GATA-1 and FOG-1 in vivo. An important open question under investigation is how recruitment of the NURD complex can lead to suppression of some genes and the enhanced expression of others. The FOG-1/NuRD mutant mice provide useful tools to dissect transcription pathways initiated by GATA-1. Moreover, given the role of GATA-1 mutations in congenital anemias and megakaryoblastic leukemias, enzymatic components of the NuRD complex may provide novel targets for pharmacologic manipulation to treat these disorders.
A Four Protein Plasma Fingerprint of Acute Graft Versus Host Disease (GVHD) Predicts Long Term Survival.
Authors: Paczesny Sophie , Choi Sung , Braun Thomas , Kitko Carrie , Oleg Krijanovski , Clouthier Shawn G. , Weyand Angela , Bickley Dan , Jones Dawn , Whitfield Joel , Levine John E. , Ferrara James L. 
Published: April 2020

Abstract
There are no independent laboratory diagnostic tests for acute GVHD. We first identified 7 potential plasma biomarkers (IL-2R-α, CRP, IL-8, TIMP-1, TNFR1, HGF, CA-19.9) of acute GVHD using a combination of proteomic approaches and antibody microarrays. We next conducted a retrospective analysis using plasma samples from 424 patients at the University of Michigan under IRB approval. We obtained samples at the first clinical signs of acute GVHD prior to treatment and at equivalent time points in patients without GVHD (Table 1). The median duration of follow-up was 420 days with a minimum follow-up of 180 days. Patients with veno-occlusive disease, idiopathic pneumonia syndrome, or septic shock were not included. We measured plasma levels of the 7 proteins by sequential ELISA. Logistic regression models with and without leave-one-out-cross-validation (LOOCV) tested the correlation of the laboratory values with the diagnosis of acute GVHD using area under the receiver-operating-characteristic (ROC) curves (AUC). The training set consisted of 282 randomly selected patients; the validation set included the remaining 142 patients. The final optimal fingerprint of four proteins excluded CRP because of its association with non-specific inflammation and included IL-2R-α, TNFR1, IL-8 and HGF, with AUCs of 0.91 and 0.89 in the training set (without and with LOOCV, respectively) and 0.86 in the validation set. The fingerprint had a strong association with grade of GVHD (p&lt;0.001) and target organ (p=0.002) at onset; interestingly, HGF had the strongest association. Using a predicted probability of acute GVHD of at least 50%, the fingerprint had a 72% sensitivity and 89% specificity. When we categorized the predicted risk of acute GVHD into low (0.00–0.59), moderate (0.60–094) and high (0.95–1.00), the plasma fingerprint predicted long-term survival (Figure 1, p&lt;0.001). We conclude that this plasma protein fingerprint has good sensitivity, high specificity, strong association with initial grade and target organ of acute GVHD, and effectively stratifies patients into three risk groups for GVHD that correlate with long term survival.
Figure Figure Table 1: Patients characteristics GVHD- (N=242) GVHD+ (N=182) Age-yr Median (range) 45 (1–69) 49 (1–71) Donor type (%) MRD: 169 (70%) MRD: 105 (58%) URD: 73 (30%) URD: 77 (42%) Conditioning regimen Intensity (%) Full: 182 (75%) Full: 114 (63%) Reduced: 60 (25%) Reduced: 68 (37%) Day after BMT of samples : median (range) 30 (7–104) 29 (5–119) Grade at GVHD Onset (%) Grade 0 Grade 1 Grade 2 Grade 3–4 242 (57%) 48 (12%) 100 (24%) 34 (7%) Organ Target at GVHD Onset (%) n/a Skin Gut Liver Combined 119 (65%) 38 (21%) 7 (4%) 18 (10%)
Susceptibility of Endocrine, Cardiac, and Macrophage Cell Lines to Iron-Mediated Oxidative Damage and the Cytoprotective Effect of the Orally Active Chelator Deferasirox (Exjade®, ICL670).
Authors: Glickstein Hava , Nick Hanspeter , Cabantchik Zvi I. 
Published: April 2020

Abstract
Systemic iron overload (primary or secondary) affects hepatic and extrahepatic functions by damaging endocrine and cardiac tissue. In vitro studies with pancreatic Min6 and pituitary Att20 cells and with cardiac H9c2 cells (all highly active in endocytotic activity) indicated that their exposure to labile iron, acutely or chronically, lead to major intracellular iron accumulation in organelles (endosomes, mitochondria, cytosol) and increased reactive oxygen species (ROS) formation when redox challenged. Among the functions affected by metal-evoked ROS are permselectivity (calcein leakage), mitochondrial ΔΨ (JC1 test), electron transport activity (Alamar Blue) and cell viability (calcein-propidium iodide). The administration at therapeutically achievable doses of deferasirox (30–100 μM) or deferoxamine (DFO) [10 μM] largely (&gt;70%) prevented labile iron from rising in cells if present in the iron-loading medium; however, only deferasirox reduced iron-evoked cell damage and increased cell viability if incubated with cells prior to or post iron-loading (acute or chronic). Thus, deferasirox has both preventive and corrective potential against iron-evoked damage in iron-loaded endocrine and cardiac cells. A gradual reduction of serum concentration from 10% (normally used in culture conditions) to 1% or less (used for experimental testing of drugs) revealed a commensurate increased susceptibility of endocrine and cardiac cells to deferasirox in the higher concentration range of 50–100 μM (48–72 cell viability test). This indicates that the plasma-binding property of deferasirox has a cytoprotective effect.
Deficient Th-1 Responses from TNFα-Matured Dendritic Cells.
Authors: Decker William K. , Li Sufang , Xing Dongxia , Robinson Simon N. , Yang Hong , Steiner David , Bollard Catherine M. , Komanduri Krishna V. , Shpall Elizabeth J. 
Published: April 2020

Abstract
INTRODUCTION: The ultimate success of dendritic cell (DC) vaccination for the active immunotherapy of neoplasia involves a wide variety of variables including the method of DC generation, loading, and maturation; the DC dose and route of administration. A maturation cocktail comprising IL-1b, TNF-a, IL-6, and PGE2 (ITIP) is commonly employed clinically, as is the use of the single inflammatory cytokine TNF-a. Here we compared these two methods with respect to DC maturation, phenotype, and function.
METHODS: CD14+ monocytes were isolated from normal donor G-CSF-mobilized peripheral blood progenitor cells using the mini-MACS system (Miltenyi Biotec), and cultured for 6 days in AIM-V medium containing GM-CSF and IL-4 (Cellgenix). Immature DCs were loaded with both acute myelogenous leukemia (AML) tumor lysate and mRNA as previously described1 and then matured with the ITIP cocktail, TNF-α alone, or a combination of TNF-α and CD40 agonism (Becton-Dickinson). Mature DCs were analyzed phenotypically and then cultured with autologous T-cells.
RESULTS: Data generated from three different normal donors indicated that DCs matured with TNF-α were deficient in important costimulatory surface molecules CD80, CD83, and CD40 as well as in IL-12 production (p=0.001 in representative experiment). Analysis of T-cell functionality by IFN-γ ELISpot also demonstrated greatly decreased Th-1 functionality among lymphocytes stimulated by TNFα-matured DCs in comparison to ITIP-matured DCs (p=0.004 in representative experiment). Additionally, lymphocytes stimulated by TNFα-matured DCs were not able to proliferate significantly whereas ITIP-matured DCs mediated robust lymphocyte expansions consisting of up to 70% CD8+ cells. The addition of a CD40 agonist antibody to TNF-α during the DC maturation process could partially, but not completely overcome these observed functional deficiencies.
CONCLUSIONS: The data suggest that DCs loaded with primary tumor antigens and then matured with TNF-a might be deficient in the induction of type 1 T-helper responses. The absence of critical costimulatory markers and IL-12 secretion suggest that DCs matured with TNF-α might be suboptimal for the priming of naïve CD8 responses. These findings might help to improve the clinical efficacy of future DC vaccination studies.
Use of Interleukin 6, High Sensitivity CRP, TNFα and Procalcitonin as Early Markers of Sepsis in Febrile Neutropenic Children.
Authors: Kharya Gaurav , Yadav Satya Prakash , Srivastava Lalit M. , Sachdeva Anupam 
Published: April 2020

Abstract
Objective: Children with cancer often develop febrile illnesses after cytotoxic chemotherapy. It is of paramount importance to determine which children have serious bacterial infections in this vulnerable period. We studied the ability of interleukin 6 (IL-6), high sensitivity C-Reactive Protein (hsCRP), tumor necrosis factor alpha (TNFα) and procalcitonin (PCT) to act as a sensitive diagnostic marker for bacterial sepsis in febrile neutropenic children and their utility as a predictive marker for serious infections.
Design: Prospective cohort study.
Setting: Sir Ganga Ram Hospital.
Patients: Sixty-two episodes in children (aged 5 months to 17 yrs) with a known malignancy/aplastic anemia who presented with fever and neutropenia.
Interventions: Serial blood samples were obtained (admission, and 48 hrs), and concentrations of PCT, IL-6, TNF α and hsCRP were determined. Demographic and laboratory data from the patients were collected from the medical records.
Measurements and Main Results: Out of 62 episodes of febrile neutropenia, fifteen (24%) of the children had proven sepsis (13 positive blood cultures, 1 invasive aspergillosis and 1 local abscess with fever). Plasma levels of IL-6, hsCRP, TNFα and PCT were increased at all time points in children with sepsis compared with those without sepsis. In culture positive patients the value of TNF-alpha was higher but statistically not significant. No major differences were found in hsCRP concentrations. IL-6 at 0 and 48 hrs were reliable markers for sepsis (area under the curve 0.73 and 0.77, respectively). Using cutoff values of IL-6 &gt; 137.5 pg/mL at 0 hrs and &gt;69 pg/mL at 48 hrs of admission produced a screening test for sepsis with 67% sensitivity, 75% specificity at 0 hrs and 75% sensitivity and 78% specificity at 48 hrs respectively. Also using the cutoff values of 1.73ng/ml for procalcitonin at 0 hrs was a good screening test with 73%sensitivity and 70% specificity. When IL-6 cutoff was brought down to 20pg/ml at 0 hr then sensitivity increased to 100%. 42/62 children with febrile neutropenia had IL6 &gt;20 which included all 15 proven sepsis cases and all 6 deaths. Rest of 20 patients all had negative cultures and no deaths. PCT value of &gt;1 ng/ml was seen in 39/62 and included 12 of 15 cases of proven sepsis. PCT was &lt; 1ng/ml in a case of invasive aspergillosis and another case of candida line infection and a case of local abscess with fever.
Conclusions: Our data shows the utility of a rapid and sensitive assay for interleukin-6 along with procalcitonin as a sensitive diagnostic marker of bacterial sepsis in febrile neutropenic children. The use of these markers as a clinical tool may allow for early discharge on oral antibiotics or no antibiotics in patients with IL6 less than 20 pg/ml and PCT less than 1 ng/ml.
Identification of Specific TCR Vβ3 and Vβ13 Genes Relate to Diffuse Large B-Cell Lymphoma-Associated Antigen.
Authors: Yin Qingsong , Li Yangqiu , Tan Huo , Chen Shaohua , Yang Lijian , Ye Jingmei , Li Bo 
Published: April 2020

Abstract
Diffuse large B-cell lymphomas (DLBL) are the most common type of non-Hodgkin’s lymphoma (NHL), account for 30% to 40% of NHL. Treatment with conventional combination chemotherapy produces complete remission rates of 50 to 70%. Target therapy such as anti-CD20 monoclonal antibody (rituximab) had shown favourable results. In recent years, specific adoptive immunotherapy with tumor-specific T-cells was considered as a new and promising treatment in malignancy. Thereby, researches are focused mainly on the generation of effective antigen specific T-cells, both autologous and allogeneic specific T cells. The antigen specific T cells could be determined by analysis of the T-cell receptor (TCR) Vα or Vβ repertoire, which display monoclonal or oligoclonal expansion. In order to develop the specific cellular immunotherapy in DLBL, in the present study, we analysis at first the distribution and clonality of TCR Vβ repertoire in peripheral blood T cells by RT-PCR and genescan technique, which was expected to define the DLBL associated TCR Vβ subfamily T cells. 6 cases with DLBL were used in the study. The skew distribution and clonal expansion of TCR Vβ subfamily T cells could be found in all patients with DLBL. Monoclonal expanded T cells were identified in 3 cases, which used the Vβ3 (1 case) and Vβ13 (2 cases) subfamilies, the PCR products from the monoclonal T cells were analyzed by direct sequencing to define CDR3 sequence. The CDR3 sequences from these 3 TCRβ genes were identified, which were not identical in the reported TCR sequences in Genebank. The whole sequences of Vβ3 and Vβ13 found in 3 patients with DLBL were amplified and cloned, which are being registered in NCBI Genebank. These specific TCR Vβ genes will be farther used for research on generation of CTL by TCR gene-modified technique, and so on. In the clinical observation showed that patients who had specific reactive T cell clones had a preferable prognosis, suggesting that these detectable clonal expanded T cells might play a crucial role in tumor eradication.
Effect of Staphylococcal Enterotoxin A on the Distribution and Clonal Expansion of TCR Vβ Subfamilies and the Cytotoxicity of T Cells Stimulated by PML-RARα Peptid.
Authors: Lin Chen , Chen Shaohua , Yang Lijian , Bai Xue , Li Yangqiu 
Published: April 2020

Abstract
Bacterial superantigens (SAgs) are potent T cell activators. Superantigens stimulate some population of T cells, based on Sag binding to specific T cell receptor variable region β chains, a property that distinguishes SAgs from conventional T cell mitogens. The biologic properties of SAgs make them attractive for use in immunotherapy. However, the T cell stimulatory effects of SAgs cannot be confined to specific antigens, its important role might be to enhance the immune effect for tumor antigens. PML-RARα peptide is a specific antigen associated with acute promyelocytic leukemia, which can induce specific proliferation of T cells from healthy human donor in vitro. But the immunogenicity of PML-RARα peptide is weak based on our previous study. In order to investigate the differentiation and proliferation of T cells induced by PML-RARα peptide that combined with Staphyloccucal enterotoxins A(SEA), T cell phenotype, TCR Vβ repertoire usage, clonality and cytotoxicity of T cells were analyzed. Peripheral blood mononuclear cells from healthy donors were cultured with PML-RARα peptide and SEA for 20 days. After induction, the distribution and clonal expansion of TCR Vβ subfamilies of T cells were examined by RT-PCR and GeneScan. The cytotoxicity of T cells induced against NB4 cell lines were examined by Cell Counting Kit-8 (CCK-8). The CD4 and CD8 surface marker on the harvested CD3+ T cells were detected by flow cytometry (FCM). And cytokine levels of IFN-γ, IL-4 and IL-10 in the cultured supernatants were measured by Enzyme-Linked Immunosorbent Assay (ELISA) as well. The results showed that restricted expression of Vβ repertoire (11 Vβ subfamilies) was found, which were Vβ1, Vβ2, Vβ3, Vβ6, Vβ7, Vβ8, Vβ13, Vβ14, Vβ16, Vβ17, Vβ19 subfamilies. Clonal expanded T cells could be identified in some Vβ subfamilies, which were preferentially used in Vβ13 and Vβ14. The specific cytotoxicity of T cells induced by PML-RARα peptide with SEA was higher than that from PML-RARα peptide alone induction group against NB4 cells. The FCM assay showed that the activation of CD8+ T cells was in priority induced by PML-RARα peptide whether with SEA or not, while it came out to be an equal between CD4+ and CD8+ T cells induced only by SEA. And the IFN-γ level of Th1-type cytokine was higher than that of the Th2-type cytokines (IL-4 and IL-8) in both cultured supernatants of T cells induced by PML-RARα peptide with or without SEA. In conclusion, PML-RARα peptide can induce specific T cells with clonal expansions of TCR Vβ subfamilies. SEA could enhance the induction of the specific PML-RARα peptide. The T cells induced by combination of PML-RARα peptide and SEA have higher specific antileukemia effect.
Effect of Mesenchymal Stem Cells on the Viability, Proliferation and Differentiation of B Lymphocytes.
Authors: Tabera Soraya , Perez-Simon Jose A. , Diez-Campelo Maria , Sanchez-Abarca Luis I. , Blanco Belen , Benito Ana , Ocio Enrique , Sanchez-Guijo Fermin M. , Canizo Consuelo , Miguel J.F. San 
Published: April 2020

Abstract
Mesenchymal stem cells (MSC) are multilineage non hematopoietic progenitor cells that play a key role in supporting the lymphohematopoietic system. Their distribution in bone marrow and secondary lymphoid organs allows an intimate interaction with T and B-lymphocytes, which contribute to the normal lymph node development, but this interaction, can not be considered as a simple bi-directional cross-talk and other cell subsets, such as dendritic cells (DC), must be considered. We have analysed the effect of MSC on B-lymphocytes and the pathways involved in these effects. For these propose, we cultured B-cell with or without MSC and analysed different markers involved in the differentiation of B-cells. We found that MSC inhibited proliferation, arresting B lymphocytes in G0G1 phase of cell cycle (figure 1). However, the presence of MSC increased the viability ob B-lymphocytes (double number of viable B-cells: from 13012 to 22835 Annexin-V PE/7 ADD negative events within B-lymphocytes in absence versus presence of MSC). The exposure of B-cells to plasmocytoid DC (pDC) induced B-lymphocytes differentiation increasing both the percentage of CD38++/CD138++ cells as well as the mean fluorescence intensity of both markers (figure 2). Accordingly, different B-cell subpopulations could be identified which represented a continuum in B-cell maturation. While the levels of cytoplasmic immunoglobulin (cIg) were higher among CD38++ cells, the opposite occurred for the expression of surface Ig as well as CD19 and CCR7. Interestingly, the presence of MSC blocked B-cell differentiation. Regarding the pathways involved in these effects, the presence of MSC influenced on ERK 1/2 and p38 pathways, but these effects depended on the culture conditions. Thus, MSC induced phosphorilation of ERK 1/2 MAPK and inhibited phosphorilation of p38 in B-cells cultured with Ig plus CpG (low proliferative conditions) while the contrary occurred in B-cells cultured with TPA (highly proliferative conditions). Therefore we demonstrated that MSC increased viability and blocked cell cycle of B-lymphocytes. Furthermore the plasmocitoid dendritic cells favoured B-lymphocytes differentiation and this process in inhibited in presence with MSC. These effects are at least in part mediated through the ERK 1/2 and p38 pahtways.
Figure Figure
Influence of mTOR Signalling Pathway on the MHC I Immunopeptidome.
Authors: Fortier Marie-Hélène , Caron Etienne , De Verteuil Danielle , Perreault Claude , Thibault Pierre 
Published: April 2020

Abstract
Cell surface major histocompatibility complex (MHC) I molecules are associated with self peptides that are collectively referred to as the self MHC I immunopeptidome (sMII). Despite the tremendous importance of the sMII, very little is known on its genesis and molecular composition. On the other hand, it is well established that the signalling pathway involving mammalian target of rapamycin (mTOR) plays an essential role in the regulation of processes such as ribosome biogenesis and protein translation which are critical for cell growth, proliferation and differentiation. In this work, we studied the influence of mTOR on the sMII for two major reasons:
the tremendous importance of this pathway in oncogenic processes its role in the control of protein synthesis, which is at the origin of the generation of the sMII. To achieve this goal, we developed a novel high-throughput mass spectrometry approach that yields an accurate definition of the nature and relative abundance of unlabeled peptides presented by MHC I molecules.
Starting from EL4 thymoma peptide extracts, more than 200 MHC I-associated peptides were sequenced with high confidence level across more than 5500 peptide clusters reproductibly identified through replicate injections (n = 3). Comparison of the sMII of EL4 thymomas before and after rapamycin stimulation revealed that 13% of the MHC I-associated peptides were significant overexpressed (fold change ≥ 2, p-value ≤ 0.05) after mTOR inhibition. Out of the 27 MHC I peptide candidates showing differential expression, 60% of peptide source proteins were linked to cell development and/or proliferation including ITFIFKSL and NAIKNHWNSTM assigned to FK506 binding protein 12 (mTOR) and myeloblastosis oncogene (c-myb) proteins, respectively. These results indicate that cell signalling events have a major impact on the composition of the sMII that can be monitored by analyses of high-throughput MS-based quantitative profiles.
Figure 1: Relative Quantification of Differentially Expressed MHC I peptides. Volcano Plot representation illustrates MHC I peptides reproductibly detected across replicate injections (n=3). Peptides over-expressed on EL4 cells after rapamycin stimulation are highlighted in blue. Figure 1:. Relative Quantification of Differentially Expressed MHC I peptides. Volcano Plot representation illustrates MHC I peptides reproductibly detected across replicate injections (n=3). Peptides over-expressed on EL4 cells after rapamycin stimulation are highlighted in blue.