bluuebunny/crossref_metadata_2025_split huggingface dataset:
Adenoviral Vector-Mediated Transfer of the Indian Hedgehog Gene Modulates Lymphomyelopoiesis In Vivo.
Authors: Kobune Masayoshi , Kawano Yutaka , Sasaki Katsunori , Takimoto Rishu , Kato Junji , Hamada Hirofumi , Niitsu Yoshiro 
Published: April 2020

Abstract
Indian hedgehog (Ihh) plays an essential role in angiogenesis, hematogenesis and epiphysis formation during embryogenesis. In the present study, we injected an adenoviral vector (Adv) carrying the mock-control (adv-control) or Ihh gene (Adv-Ihh) into SCID or BALB/c mice to evaluate the effects of lhh on the regulation of postnatal hematopoiesis in vivo. After the intravenous injection of Adv-Ihh, the expression of vector-derived Ihh mRNA was detected in the liver. Four weeks after administration of Adv-Ihh to SCID mice, we observed an increase in the number of c-Kit+ cells and clonogenic cells/105 mononuclear cells in the bone marrow as compared with adv-control administrated mice. Moreover, after administration of Adv-Ihh to BALB/c mice, the number of splenic B220+IgMlowCD23intCD21int B lymphocytes (Figure 1) and CD4+ T lymphocytes (Figure 2) was strongly increased. Furthermore, the number of thymic double negative (DN)2, DN3, CD8+ immature single positive and CD4+/CD8− cells was significantly elevated relative to the number in mice that received the control Adv vector. Our results suggest that enhanced signaling by Ihh can modulate the proliferation and differentiation of splenic B lymphocytes and thymic T lymphocytes during BM hematopoiesis in vivo. Thus, modulation of the Hh signaling pathway may provide a therapeutic strategy to stimulate lymphomyelopoiesis in vivo.
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Expression of miR-188 and 362 Induced by Erythropoietin Stimulation in a Human Erythrocytic Leukemia Cell Line.
Authors: Sugiura Keiichi , Kosaka Nobuyoshi , Yamamoto Yusuke , Yoshioka Yusuke , Miyazaki Hiroshi , Komatsu Norio , Ochiya Takahiro , Kato Takashi 
Published: April 2020

Abstract
MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression at the post-transcriptional level and participate in a lot of biological processes. Several studies indicated that miRNAs play an important role in hematopoietic system. It has not been elucidated well about miRNAs involved in cytokine-regulated molecular mechanisms in cellular proliferation and differentiation. We screened specific miRNAs expressed in human leukemia cell lines, UT-7 and its sublines UT-7/GM, UT-7/EPO and UT-7/TPO, which exhibit hematopoietic lineage properties depending on cytokine stimuli by GM-CSF, erythropoietin (EPO), or thrombopoietin (TPO), respectively. The expression profiles of 470 miRNAs in all were analyzed using LNA-based microarray between UT-7/GM, UT-7/EPO and UT-7/TPO cells. The initial microarray data were validated, and specific expressions of several miRNAs were confirmed using quantitative real-time PCR. Especially, the expression levels of miR-188 and 362 clustered on X chromosome were significantly higher in UT-7/EPO cells as compared with those in UT-7, UT-7/GM and UT-7/TPO cells. To assess the effect of EPO on the expression of miR-188 and 362, UT-7/EPO cells were cultured under various concentration of EPO (0 to 10 U/ml) after cytokine withdrawal for 24 hours. Quantification of miRNAs expression by real-time PCR showed that miR-188 and 362 were increased 2- to 20-fold by EPO stimuli, but both expressions were not in dose-dependent manner. Furthermore, the addition of EPO (1 ng/ml) to growth factor-derived UT-7 cells which were cultured for 1 month caused a 4- to 50-fold increase of miR-362 expression; while miR-188 expression was not increased. Similarly, EPO induced the expression of miR-362 in erythroleukemic cell line TF-1. The expression levels of miR-188 and miR-362 were significantly higher in UT-7/EPO with erythrocytic features than UT-7/GM and UT-7/TPO cells, in addition, expressions of both miRNAs were induced by EPO stimuli in UT-7/EPO cells, suggesting that miR-188 and miR-362 are involved in lineage specific molecular mechanisms of erythropoiesis.
Impaired Repopulating Activity of Fus-Deficient Hematopoietic Stem Cells.
Authors: Sugawara Takeaki , Iwama Atsushi 
Published: April 2020

Abstract
RNA-binding protein FUS (also known as TLS) was originally identified in chromosomal translocation in human myxoid liposarcoma. The FUS gene is also translocated with the transcription factor gene ERG in human myeloid leukemia with recurrent chromosomal translocation t(16;21). Multiple data suggest that wild-type FUS is also involved in the development of leukemia as one of the downstream targets for oncoproteins including BCR-ABL. However, little is known about the role of FUS in the normal hematopoiesis. The previous report demonstrated that Fus-deficient (Fus−/−) newborn mice, which die shortly after birth because they cannot suckle, have a non-cell-autonomous defect in B lymphocyte development. No cell-autonomous defect of Fus−/− hematopoietic cells has been documented. Here we report the detailed analyses of the Fus−/− fetal liver hematopoietic stem cells (HSCs). Fus−/− fetal livers at embryonic day 14.5 were smaller in size and exhibited a significant reduction in hematopoietic cell numbers by 60% compared with the wild type (WT). Nonetheless, no significant difference was observed in the proportion of stem/progenitor cell fraction (lineage-marker-c-Kit+Sca-1+; KSL) as well as colony-forming cells between WT and Fus−/− fetal livers. Fus−/− KSL cells proliferated and differentiated almost normally in vitro. To examine in vivo repopulating activity, we transplanted fetal liver cells to lethally irradiated CD45.1 recipients with competitor bone marrow cells. Fus−/− fetal liver donor cells reconstituted recipients’ hematopoiesis for the long term and contributed to all cell lineages including B lymphocytes. In contrast to the in vitro results, however, the chimerism of donor-derived cells was significantly lower in recipients receiving Fus−/− fetal liver cells compared with WT controls (approximately 2-fold reduction). This trend was reproducible with both unfractionated and purified KSL fetal liver test cells. Our data demonstrated that the proto-oncogene Fus is involved in the maintenance of normal HSC functions. Detailed analyses on the underlying mechanisms are in progress.
Functional Interaction of c-Kit Receptor and Interleukin-3 Receptor.
Authors: Ye Zhijia , Weissman Sherman M. 
Published: April 2020

Abstract
Hematopoiesis is regulated by a number of growth factors and cytokines, among which stem cell factor (SCF) plays a critical role for normal hematopoiesis. An important feature of SCF is its strong capacity to synergize with the cytokines including interleukin-3(IL-3), granulocyte-marcrophage colony-stimulation factor (GM-CSF) and erythropoietin to regulate proliferation and differentiations of hematopoietic progenitor cells. The mouse hematopoietic progenitor cell line EML contains two populations of lineage negative cells, distinguished by the expression or absence of CD34. The two sub-populations express similar levels of the SCF receptor c-kit, but only the CD34+ population replicates in the presence of SCF alone, while the CD34 negative population replicates in the presence of IL3 but this replication is synergistically stimulated by the addition of SCF. Our previous studies indicated that synergistic proliferation of the mouse hematopoietic progenitor cell line EML induced by SCF in combination with IL-3 was probably mediated by the synergistic increases in tyrosine phosphorylation of c-Kit and β-chain of IL-3 receptor, which is shared with GM-CSF receptor and interleukin-5 receptor. Conversely, the decreased response of c-kit to SCF alone in the CD34 negative population may be a result of the increased expression of IL3 receptor in these cells. Trans-phosphorylation between c-Kit and the β-chain of IL-3 receptor was also observed in EML cells. In studies of the molecular mechanism behind the functional interactions between SCF and IL-3 we found that c-Kit and IL-3 receptor β-chain form a complex in EML cells. Antibody Ab-1 specific against the Ig-like domain 4 in the extracellular region of c-Kit blocked not only cell proliferation induced by SCF but also synergistic proliferation induced by SCF plus IL-3. Consistent with this finding, Ab-1 inhibited phosphorylation of c-Kit induced by SCF or IL-3, and synergistic phosphorylation of c-kit induced by SCF plus IL-3 respectively. SCF and IL-3 in combination synergistically activated the protein kinases Akt and Erks, the downstream mediators of c-Kit and IL-3R. Phosphorylation of Erks is independent on the phosphatidylinositol 3-kinase (PI3-kinase) pathway in EML cells. These data suggested the possibility that the synergistic growth of EML cells induced by SCF in combination with IL-3 was mediated by multiple mechanisms that could include trans- and synergistic phosphorylation of c-Kit and the β-chain of IL-3 receptor, PI3-kinase dependent synergistic phosphorylation of Akt and PI3-kinase independent synergistic phosphorylation of Erks. The trans- and synergistic phosphorylation of c-Kit and β-chain of IL-3 receptor was mediated by physical interaction of two receptors.
MicroRNA Analysis by In Situ Hibridization in Hodgkin Lymphoma.
Authors: Navarro Alfons , Martinez Antonio , Balagué Olga , Gaya Anna , Pons Aina , Urbano-Ispizua Alvaro , Montserrat Emili , Monzo Mariano 
Published: April 2020

Abstract
Hodgkin lymphoma (HL) is a neoplasm characterized by the presence of relatively few tumoral cells, Hodgkin and Reed-Sternberg cells in an important non-neoplastic environment. The mature microRNAs (miRNAs) are small RNA molecules (20–25 nt), that act inhibiting the mRNA translation to protein by binding to 3′ UTR region of mRNA. In a previous work we detect a 25 miRNA signature of HL. Some of them inhibit the expression of key genes related to B lymphomagenesis. The aim of the study was to determine if the differential expression of miRNAs in tumoral tissue versus reactive lymph nodes, is due to genetic alterations in the Hodgkin/Reed Sternberg cells or alterations in the non-tumoral microenvironment. Fluorescein (FITC) 5′ labelled locked-nuclei-acid-incorporated (LNA) miRNA ribo probes for miR-21, 134, 138, 155 (miRCURY™ LNA detection, Exiqon) were used in 20 cases of HL formalin-fixed paraffin embedded tissue sections on silane coated slides (Vision BioSystem). Chromogenic in situ hybridization was done in an automated platform Bond Max (Vision Biosystems) with minor modifications. Pre-treatment of the slides was performed with Protease 1 for 10 min at 37°C. A total amount of 300 microliter of 25nM probe was hybridized in 1x sodium chloride-sodium citrate hybridization buffer (SSC) (Innogenetics) up to 50° C for 2 hours. We used a pre-diluted mouse anti-FITC antibody (Vision BioSystems) for 20–60 minutes followed by a goat anti-mouse linked to thousands of horse radish peroxidase (HRP) sites (Refine Detection System, Vision BioSystems). DAB was used as a chromogen reacting for 10 minutes and hematoxilyn was used as a counterstain. By functional and target analysis, we selected three miRNAs, miR-21 (PTEN), miR-134 (J-Chain) and miR-138 (PU.1), from the cHL signature that seemed to have a role in tumorigenesis process and analyzed them by CISH. In all cases a cytoplasmic signal was demonstrated in Hodgkin and Reed-Sternberg cells. Moreover, a nuclear signal was identified in reactive tumor infiltrating lymphocytes for miR-21 and miR-138. This nuclear signal may be due to crossreactivity with primary miRNA in these cells. We analyzed miR-155 as positive control of in situ hibridization and we demonstrated a cytoplasmic signal in Hodgkin and Reed Sternberg cells, as well as in scattered reactive lymphocytes and activated histiocytes as previously reported. In conclusion, miR-21, miR-134 and miR-138 play a role in HL lymphomagenesis since their expression is preferentially localized in Hodgkin/Reed Sternberg cells. The study of the other miRNAs of our signature can help explain the changes that appear in the atypical cells as well in the reactive cellular microenvironment in HL.
The Association between Hepatitis C Infection and Lymphoma in AIDS.
Authors: Ghanem Zuhair , Singh Amandeep , Zahran Ali , Maroules Michael 
Published: April 2020

Abstract
Background: Many studies have demonstrated a possible association between hepatitis C and a wide spectrum of lymphoproliferative malignancies. Increased hepatitis C virus (HCV) prevalence among patients with non-Hodgkin lymphoma (NHL), suggesting that HCV infection increases NHL risk through chronic immune stimulation. Most of the previous studies excluded patients with immunodeficiency. No study looked at the association between established chronic HCV infection and lymphoma in AIDS patients. However, previous studies found no relation between HCV infection and lymphoma in HIV patients. The aim of our study is to determine the correlation between chronic HCV infection and lymphoma in AIDS patients.
Methods: A hospital based retrospective case control study was performed in a tertiary care center. The medical records of 603 AIDS patients were reviewed between January 2001 and December 2005. 284 patients with both AIDS and hepatitis C in the first group and 319 control patients with AIDS only in the second group. The diagnosis of hepatitis C and AIDS was determined retrospectively from the patients medical records. Only patients with AIDS were included in the study. AIDS was defined as CD4 &lt; 200 or any AIDS defining illness.
Results: Among the 284 patients with both AIDS and HCV infection 7 (2.4%) patients were found to have lymphoma at any stage of the disease. On the other hand, 8 (2.4%) patients were found in the control group. By using the Fisher Exact Probability Test there was no statistically significant increase in the incidence of lymphoma among AIDS patients with HCV infection compared to the control group (p=0.823063, and Odds Ratio =0.9824). The type of lymphoma was not determined giving the small number of cases and that the majority of lymphoma in AIDS are NHL cases.
Conclusion: Our data showed that among AIDS patients, co infection with Hepatitis C virus doesn’t increase the incidence of lymphoma. Giving the fact that AIDS is a low immunity status, this conclusion supports the theory that chronic immune stimulation plays a role in the pathogenesis of lymphoma in HCV infection.
Interleukin-15 Upregulates the Expression of NK-Cell Activating Receptors and Concomitantly Increases the NK Cytotoxicity in Patients with Acute Myeloid Leukemia.
Authors: Szczepanski Miroslaw J. , Czystowska Malgorzata , Szajnik Marta , Welsh Ann , Foon Kenneth A. , Whiteside Theresa L. , Boyiadzis Michael 
Published: April 2020

Abstract
Natural killer (NK) cells lyse malignant cells without prior antigen-specific priming and play a critical role in the innate immune response. A balance of signals from activating and inhibiting receptors expressed on each NK cell controls its activity. The growth, differentiation and survival of NK cells have been found to be dependent on interleukin-15 (IL-15). Using multicolor flow cytometry we investigated the receptor repertoire and also measured the NK cell activity in twenty three patients with newly diagnosed acute myeloid leukemia (AML) prior to any treatment. Further, we investigated the ex-vivo effect of IL-15 on the NK cell repertoire and NK cell cytotoxicity. The percentage of circulating NK cells was lower (p&lt;0.0001) in the AML patients (6%± 0.7, range 1–17%) compared to the NK cells of healthy donors (12%± 1, range 9–17%). The expression of the activating natural cytotoxicity (NCR) receptors NKp30 and NKp46 and the C-type lectin receptors NKG2D and NKG2C was significantly decreased in the AML patients compared to the NK cells of healthy donors: NKp30 24 vs 51% p&lt;0.0001, NKp46 32 vs 73% p&lt;0.0001, NKG2D 43 vs 83% p&lt;0.0001, NKG2C 17 vs 28% p&lt;0.03. In addition, the receptor expression (mean fluorescence intensity, MFI) was also significantly lower in AML patients compared to healthy donors. No significant differences in the expression of the NCR NKp44 and the NK- cell inhibitory receptors were observed. Furthermore, the NK cytotoxicity in the AML patients at diagnosis was significantly lower (p&lt;0.0003) compared to the NK cytotoxicity of healthy donors (4 vs 75 LU). When NK cells obtained from AML patients were cultured with IL-15, significant increases in the expression of the NK- cell activating receptors (Table 1) were observed. The upregulation of the activating receptors was associated with a concomitant significant increase (p&lt;0.001) of the NK cell cytotoxicity (4 vs 70 LU). The data suggest that IL-15, a homeostatic NK cell cytokine, can upregulate the expression of activating receptors and concomitantly increase the NK lytic activity. The use of IL-15 as a platform for NK- based therapies for AML patients should be considered in the future.
Table 1. The effect of IL-15 on the receptor expression in AML patients NK cell Activating Receptors NK cells in AML pts at diagnosis NK cells in AML pts after IL-15 stimulation p value % positive, MFI % positive, MFI NKp30 24, 2.6 84, 6.0 0.001 NKp44 4, 2.4 71, 6.7 0.001 NKp46 32, 2.6 83, 7 0.002 NKG2C 17, 2.2 58, 6.9 0.005 NKG2D 43, 2.5 87, 7.8 0.01
hDectin-1 Mediates Cross-Presentation of Epitopes Via the Cytosolic Pathway.
Authors: Grünebach Frank , Weck Markus M. , Appel Silke , Werth Daniela , Sinzger Christian , Bringmann Anita , Kanz Lothar , Brossart Peter 
Published: April 2020

Abstract
Human (h)Dectin-1 is a member of the C-type-lectin-like receptor family that was shown to be the major receptor for fungal β-glucans and to play an important role in cellular responses mediated by these carbohydrates. It is mainly expressed on human DCs and macrophages. In our study, we observed that activation of monocyte-derived dendritic cells (MDCs) with TLR3 ligand (poly I:C) but not with TLR ligand 7/8 (R848) resulted in down-regulation of hDectin-1 expression and this down-regulation correlated with a reduced uptake of apoptotic cells in phagocytosis assays. In order to analyze the possible cross-presentation of engulfed antigens we used CMV infected human fibroblasts (HFF). We found that hDectin-1 is involved in the uptake of CMV-infected HFF leading to cross-presentation of CMV-derived peptides on MHC class I molecules and activation of CMV pp65-specific CD8+ T-lymphocytes. To further delineate the pathway leading to presentation, we pretreated MDCs with compounds that inhibit processing of antigens at defined steps during presentation. Cytosolic protein degradation is performed by the proteasome, a large multicatalytic protease complex. Lactacystin specifically inhibits the 20S and 26S proteasome activity by targeting the catalytic subunit. In standard 51Cr-release assays, addition of lactacystin completely inhibited the presentation of pp65-derived peptides indicating the involvement of the proteasome in these process. The fungal product brefeldin A blocks the MHC class I processing pathway by specifically inhibiting the vesicular egress from the ER and the Golgi complex. In line with previous findings, incubation with brefeldin A almost completely abolished the lysis of MDCs incubated with CMV+ HFF. To further analyze whether the cross-presentation of CMV-derived peptides on HLA class I molecules was dependent on lysosomal proteases, MDCs that were co-incubated with HFF as above were treated with the lysosomotropic agent chloroquine that prevents acidification of the lysosomal compartment involved in the exogenous pathway of antigen presentation. The addition of chloroquine had no effect on the cross-presentation of CMV-derived epitopes on HLA class I-molecules. Summarized, the data reported here show that hDectin-1 functions not only as a pattern recognition receptor in innate immunity but is also involved in the clearing of apoptotic cells and cross-presentation of cellular antigens on MHC class I molecules to specific CTLs.
Enhancement of the Antigen-Presenting Cell Function of Mantle Cell Lymphomas (MCL) by Novel Molecularly Based Immunotherapeutic Strategies.
Authors: Wang Hongwei , Cheng Fengdong , Noyes D. , Wright K. , Mohapatra S. , Tao J. , Schell M. , Sebti S. , Smith M. , Bhalla K. , Pinilla J. , Dessureault S. , Celis D. , Sotomayor E.M. ,   (Intr. by Eduardo M. Sotomayor)
Published: April 2020

AbstractMCL is an aggressive and incurable B-cell malignancy with an intrinsic characteristic to relapse after an initial good response to treatment. Manipulation of the immune system to unleash its well-known specificity and long-lasting protective effect might provide a unique opportunity to induce more durable responses in MCL. In previous studies in an A20 B-cell lymphoma murine model we have demonstrated that augmentation of the antigen-presenting cell function of the malignant B-cell is required for elicitation of an effective anti-lymphoma immunity1. Inhibition of Stat3 signaling, a negative regulator of inflammatory responses, and modulation of histone deacetylases function were identified as two novel targets to augment the immunogenicity of the malignant B-cell. In this study we determined therefore whether manipulation of these intracellular pathways in murine and human MCL cells could result in priming of antigen-specific T-cells and/or restoration of the responsiveness of tolerant T-cells. First, in vitro treatment of FC-muMCL1 cells - cell line derived from a MCL tumor that arise following pristine injection into Em-cyclin D1 transgenic mice- with increasing concentrations of the Stat3 inhibitors, Cucurbitacin I (CuI) or CPA-7 resulted in an enhanced presentation of OVA-peptide to naive CD4+ T-cells specific for a MHC class II restricted epitope of Ovalbumin (OT-II cells). Indeed, these antigen-specific T-cells produce higher levels of IL-2 and IFN-gamma compared to anti-OVA T cells that encountered cognate antigen in FC-muMCL1 cells treated with LPS alone. Similarly, we found that culture of the human MCL cells JEKO or Z138 with allogeneic human peripheral blood mononuclear cells in the presence of increasing concentrations of the Stat3 inhibitors also resulted in increased IL-2 production by T-cells. In the next set of experiments, we determined whether treatment of FC-muMCL1 cells with the hydroxamic acid analogue pan-HDAC inhibitor LAQ824 could influence their antigen-presenting capabilities and their ability to activate T-cell responses. Unlike, MCL cells treated with LPS alone, FC-muMCL1 cells treated with LPS and LAQ824 effectively prime antigen-specific CD4+ T-cells as determined by their production of both IL-2 and IFN-gamma in response to cognate peptide. Furthermore, in vitro treatment of Z138 MCL cells with LAQ824 also led to enhanced IFN-gamma production by allogeneic human PBMCs. Taken together, our findings points to inhibition of Stat3 signaling and inhibition of histone deacetylases as appealing molecularly based immunotherapeutic strategies to augment the immunogenicity of MCL cells.