bluuebunny/crossref_metadata_2025_split huggingface dataset:
Transforming Growth Factor-β1 Inhibits the Maturation of Dendritic Cells Via Toll-Like Receptor 4 Signaling Pathway.
Authors: Mou Haibo , Lin Maofang , Huang He 
Published: April 2020

Abstract
We have demonstrated that transforming growth factor-β1(TGF-β1) inhibits the maturation of mouse bone marrow derived dendritic cells (DCs). TGF-β1 treated DCs (TGFβ-DCs) are resistant to maturation stimulus -lipopolysaccharide (LPS) and might have some correlation with the down-modulation of Toll-like receptor 4 (TLR4) expression. It was known TLR4 binds LPS from Gram-negative bacteria, triggering signaling pathways that lead to the activation of NF-κB, ERK1/2 and p38 MAPK and ensuing gene expression of proinflammatory factors. In the current study, we further estimated the activities of NF-κB, ERK1/2 and p38 proteins involved in TLR4 signaling pathway. Using EMSA method, we found the NF-κB DNA binding activity in immature DCs (imDCs) was significantly increased in response to LPS, but addition of TGF-β1 to DCs inhibited NF-κB binding. Moreover, TGF-β1 was effective in suppressing LPS-induced activation of ERK1/2 and p38 kinase, the level of phosphorylation of ERK1/2 and p38 kinase were lower than imDCs measured by Western Blot. After treatment of imDCs and TGFβ-DCs with LPS for 24 h, the production of IL-12p70 of TGFβ-DCs was significantly less than that of imDCs(115.4±15.2 pg/ml vs 517.0±29.7 pg/ml, P&lt;0.01), but the level of Th2 cytokine-IL-10 was elevated(132.1±17.5 pg/ml vs 75.1±16.6 pg/ml, P&lt;0.05), indicating that exposure to TGF-β1 impaired the capability of DCs to produce high amounts of bioactive IL-12p70. According to the semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), the expressions of chemokines MIP-1α mRNA on TGFβ-DCs after LPS stimulation were lower than imDCs at each time point. The lower expressions of MCP-1 and IP-10 on TGFβ-DCs at irregular pattern after LPS treatment, whereas the expressions RANTES were no different. Hence, the results suggested TGF-β1 maybe directly inhibit TLR4 expression on DCs, and then interfere with the activity of downstream key proteins, such as NF-κB, ERK1/2 and p38. Ultimately, TGF-β1 treated DCs were resistance to LPS, down-regulated the expression of costimulatory molecular on DCs and decreased the secretion of inflammatory cytokines.
Identification of Chitotriosidase Isoforms in Plasma of Gaucher Disease Patients by Two Dimensional Gel Electrophoresis.
Authors: Quintana Lucia , Monasterio Alberto , Escuredo Kepa , Amo Jokin del , Alfonso Pilar , Elortza Félix , Santacruz Simon , Simon Laureano , Martinez Antonio , Pocovi Miguel , Castrillo Jose L. , Giraldo Pilar 
Published: April 2020

Abstract
Introduction: Chitotriosidase protein (CT) is the most important biochemical marker described for Gaucher disease (GD). CT activity is increased several hundred-fold in plasma of GD patients and shows a strong positive correlation with the severity of the disease. However, a recessively inherited enzyme deficiency, with an incidence of about 6% in the Caucasian population, means that not all patients with GD can be monitored by measuring CT activity.
Materials and Methods: This study was performed with plasma samples from 22 type 1 GD patients and 23 healthy subjects. All patients are from Spanish GD Registry. Applying two dimensional gel electrophoresis (2-DE) we compared the plasma proteome profile of GD patients with that of healthy subjects. Differential proteins were excised and prepared for MALDI-TOF analysis. Results were verify with 2-DE western blot.
Results: In this study we have described five CT isoforms in a 2-DE gel from GD plasma samples. We show and compare the 2-DE images of each CT spot in plasma protein samples from healthy controls, GD patients with wild-type CT protein, and GD patients either homozygous or heterozygous for the 24 bp duplication. We propose a connection between the quantitative image analysis of each CT spot and the enzymatic activity of CT in each patient. Lastly, our results verifies that post-translational glycosylation explains at least some of differences between CT isoforms.
Conclusion: The possibilities of proteomic technology in the investigation of biological biomarkers of GD and the nature of the CT protein are demonstrated, as well as in detecting new biochemical markers useful for the diagnosis of the disease and for monitoring patient response to therapy.
Characteristics of Epstein-Barr Virus-Associated Hemophagocytic Lymphohistiocytosis (HLH) in European Children.
Authors: Wagner Hans-Joachim S. , Beutel Katrin , Schneider Marion E. , Janka Gritta E. 
Published: April 2020

Abstract
Hemophagocytic lymphohistiocytosis (HLH) represents a severe sepsis-like disease with massive hypercytokinemia and with proliferating and organ-infiltrating phagocytes leading to cytopenia of different hematopoietic lineages. Besides primary familial cases of HLH, infections can cause secondary forms of HLH, among which Epstein-Barr virus (EBV) infections are the most frequent trigger. Between 2003 and 2005, 10 cases of EBV-associated HLH were analysed at a local subcenter of the international HLH study group. Serial blood samples from these 4 boys and 6 girls (age range: 2–14 years, median of age: 4 years) were examined for EBV load in peripheral blood mononuclear cells (PBMC) and plasma by real-time PCR analysis. EBV loads in both PBMC and plasma were significantly elevated in all HLH patients as compared with healthy controls and also with immunocompetent patients suffering from primary EBV infections. In one patient with excessively elevated EBV load in peripheral blood, analysis of EBV-DNA in separated T and B cells by real-time PCR showed an EBV infection of both cell types. During the course of disease, EBV load in HLH patients reflected the activity of disease. Regarding the clinical course and biochemical markers of disease such as LDH, ferritin or soluble IL2-receptor levels in plasma, patients with EBV-associated HLH did not differ from other patients with HLH. Conclusion: Measurement of EBV-load by real-time PCR facilitates detection of EBV-associated cases of HLH and monitoring of affected patients during the course of disease. EBV load analysis should become part of the initial routine examinations of newly diagnosed HLH patients in order to identify the frequency of EBV-associated forms of HLH. Without molecular diagnosis by means of EBV load measurement in peripheral blood, EBV-associated cases of HLH may be underdiagnosed in non-Asian populations.
Involvement of Fps/Fes and Fer Kinases in c-Kit Signalling in Mast Cells and Mast Cell Leukemias.
Authors: Samayawardhena Lionel , Craig Andrew W.B. 
Published: April 2020

Abstract
The c-Kit receptor protein-tyrosine kinase (PTK) is required for mast cell differentiation and function. Signalling by c-Kit normally depends on binding its ligand, Stem Cell Factor (SCF). However, several constitutively activating mutations in c-Kit have been described in myeloid leukemias (including mast cell leukemias (MCLs)) and in solid tumors. The objective of this study is to define the involvement of two non-receptor PTKs called Fps/Fes (hereafter referred to as Fps) and Fer in c-Kit signaling in mast cells and MCLs. Recent studies using transgenic mouse strains bearing kinase-inactivating or null mutations in Fer and Fps have revealed roles in hematopoiesis, and in limiting the innate immune response. Recently, we showed that Fer and Fps are activated downstream of the high affinity IgE receptor, FcεRI, in bone marrow-derived mast cells (BMMCs), and that Fer kinase is required for sustained p38 Mitogen-activated protein kinase (MAPK) activation and chemotaxis upon activation of either FcεRI or c-Kit.
In this study, we focus on the roles of Fer and Fps in c-Kit signaling in BMMCs. SCF-induced c-Kit activation led to rapid association of both kinases with c-Kit, and increased tyrosine phosphorylation of Fer and Fps. Interestingly, kinase-dead Fps protein was also phosphorylated upon c-Kit activation, suggesting the involvement of an upstream kinase. Pretreatment of BMMCs with the Src family kinase inhibitor SU6656, caused reduced phosphorylation of Fer and Fps upon SCF treatment. This suggests an involvement of one or more Src family kinases (SFKs) in phosphorylating Fps and Fer upon c-Kit activation. We next examined the potential involvement of the Fyn SFK, since it is known to interact with a juxtamembrane site in activated c-Kit and has been linked to Fer activation in other cell types. SCF treatment of BMMCs from Fyn knock-out mice (fyn-null) revealed normal activation of c-Kit, Erk MAPK, but reduced phosphorylation of Fer kinase. Furthermore, we have observed reduced phosphorylation of Shp2 phosphatase, and the p38 and Jnk MAPKs in Fyn-null BMMCs compared to control. We are currently attempting to restore Fyn expression by retroviral transduction to attempt to rescue these signalling defects, and also evaluate potential requirements for Fyn in chemotaxis and cytokine production. Using MCL cell lines harbouring constitutively actived c-Kit (RBL-2H3 and HMC-1), we have observed that phosphorylation of Fer and Fps kinases are inhibited by treating cells with either a c-Kit inhibitor (SU11652) or SFK inhibitor (SU6656).
In conclusion, Fer and Fps kinases participate in c-Kit signaling in normal and leukemic mast cells, and Src family kinases likely contribute to their activation. Using mast cells from Fer/Fps-deficient mice, we will identify their downstream targets and determine if they are required for leukemogenesis by an activated c-Kit transgene.
Modulation of DNA Binding Properties of CCAAT/Enhancer Binding Protein Epsilon by Heterodimer Formation and Interactions with NFkappaB Pathway.
Authors: Chumakov Alexey M. , Silla Agness , Williamson Elizabeth A. , Koeffler Phillip H. 
Published: April 2020

Abstract
Specific regulation of gene expression is achieved through interaction of multiple transcriptional activators and repressors with their cognate DNA sites. This activity is subject to several levels of control, including post-translational modification by phosphorylation, acetylation, selective degradation and interaction with co-activators and co-repressors.
Recent studies directed at discovering C/EBP- responsive genes resulted in identification of a diverse and degenerate collection of putative C/EBP binding sites. Identification of binding sites for each individual member of the C/EBP family is complicated by common co-expression of several of these proteins in the same cell and their ability to form hetero-dimeric complexes with each other and structurally-related ATF/CREB proteins. Activity of C/EBP proteins is regulated by phosphorylation mediated through several major kinase pathways, that affect DNA-binding, subcellular localization in the nucleus, and interaction with cell cycle proteins.
Our studies of C/EBP-e phosphorylation identified several sites, including one for p38 MAPK within the transactivation domain of C/EBP-e. As a result of phosphorylation at Threonine-75 (T75), C/EBP-e DNA binding and specific transcriptional activity was significantly enhanced.
In this study, we identified consensus binding sequences for C/EBP-e and for heterodimers of C/EBP-e with its most common dimerization partner ATF4. Furthermore, we find that C/EBP-e- DNA interaction is highly upregulated by interaction with activated NFkappaB pathway protein p65-RelA. This interaction depends on the T75 phosphorylation status of C/EBP-e and can be enhanced by introduction of phosphomimetic Thr to Asp mutation. By using selective removal of p65-RelA and siRNA mediated p65 knock-down, we demonstrate that activated NFkappaB pathway is required for high level expression of genes with C/EBP-e responsive promoter. This novel interaction may play an important role in regulation of a C/EBP-e - dependent genes in myeloid cells, for example at sites of infection or other inflammations.
Feline Immunodeficiency Virus Depletes Naïve (CD45RA+) CD4+ Lymphocytes from the Blood and Lymph Nodes of Domestic Cats during Acute Pediatric Infection.
Authors: Johnson Calvin M. , Mergia Ayalew , Novak Janelle , Gengozian Nazareth 
Published: April 2020

Abstract
Feline immunodeficiency virus (FIV) is an immunosuppressive lentivirus of domestic cats that serves as an animal model for the pathogenesis of CD4+ lymphopenia and thymus dysfunction in HIV infected humans. During most cases of adult and pediatric HIV infection, naïve CD4+ T lymphocytes recognized by the expression of the RA isoform of the leukocyte common antigen (CD45RA) are infected at a lower level than memory CD4+ T− lymphocytes; however, children with rapidly progressive disease due to thymic insufficiency harbor high levels of HIV within the CD45RA+ subpopulation. In FIV infected cats, the fate of naïve CD4 lymphocytes is unknown due to the lack of specific markers. Recently, a mAb (755) was reported to recognize the feline homologue to CD45RA, allowing the enumeration of naïve CD4 and CD8 lymphocytes in cats. The purpose of this study was to characterize the fate of CD4+CD45RA+ blood cells eight weeks after FIV infection. One-day-old kittens (n=6) were infected with virions either from a wild type clone (JSY3) or mutant ORF-A clone at equivalent reverse transcriptase units and compared to historical control data. Eight weeks after inoculation, the percentages of CD4+ and CD8+ cells belonging to the CD45RA+ subpopulation were measured by two-color flow cytometry. Both FIV inocula were associated with a reduction in total CD4+ lymphocytes from a median of 13% in controls to 8% in infected cats (P=0.004), contributing to a reduction in the CD4:CD8 ratio from 2.45 in controls to 0.76 in infected cats (P=0.007). The decline in CD4+ lymphocytes was attributable to a disproportionate loss of CD4+CD45RA+ cells: 69% of CD4+ cells were CD45RA+ in controls, as compared to 7% in FIV infected cats (P=0.004). In contrast, naïve CD8+ lymphocytes did not change significantly with FIV infection (67% of CD8+ cells were CD45RA+ in FIV infected cats as compared to 80% in controls). The distribution of CD45RA+ cells in the lymph nodes of FIV infected cats mirrored those in the blood. Together, these data suggest that acute FIV infection results in a rapid depletion of naïve CD4 lymphocytes throughout the blood and secondary lymphoid tissues, while proportions of naïve CD8 lymphocytes remain unchanged. CD4+CD45RA+ cells may be depleted during pediatric FIV infection through lytic infection or a transition to a memory phenotype lacking CD45RA.
Compound Heterozygeous RAG-1 Mutations in 2 Patients Presenting with Chronic Disfiguring Granulomatous Skin Lesions.
Authors: Schuetz Catharina , Gudowius Sonja , Niehues Tim , Schulz Ansgar , Schwarz Klaus , Debatin Klaus-Michael , Friedrich Wilhelm 
Published: April 2020

Abstract
Introduction
Mutations of the recombinase activating genes (RAG) cause a severe combined immunodeficiency (SCID) characterized by absent or low T and B cells due to defective T/B cell receptor rearrangement and failure of lymphocyte maturation. Presenting clinical features commonly are failure to thrive and opportunistic infections in early infancy. We observed 2 patients with RAG mutations where partial T- and B-cell immunity is preserved. These patients did not suffer from severe infections but presented with unusual symptoms and signs, e.g. unexplained lymphogranulomatous skin lesions mimicking malignant lymphoma and lupus vulgaris.
Case presentation
Both patients, who are offsprings of unrelated German parents, underwent genetic screening because of unclear primary immunodeficiencies and were found to have compound heterozygeous RAG-1 mutations (Patient 1: R507W, R737H; R314W and patient 2: R778Q; R975W). Both have no evidence of thymic tissue on ultrasound and have persistant lymphopenia with low T-cell numbers, but are capable of mounting positive antibody responses to tetanus toxoid. In both patients unusual skin lesions were leading clinical abnormalities.
Patient 1, a 6 year-old girl, has been suffering from progressive papulonodular skin lesions of extremities and face since the age of 1 year. Biopsy showed highly monomorphous T-cell infiltrates leading initially to a diagnosis of cutaneous T-cell lymphoma treated with chemotherapy. As progression occurred chemotherapy was stopped and the lesions were reinterpreted as poorly organised sarcoid-like granulomas of unknown etiology. At 5 years of age the girl developed an EBV lymphoma of her right tonsil treated with rituximab. The patient is undergoing at present hematopoetic stem cell transplantation (HSCT).
Patient 2 is an 8 year-old girl, who at the age of 2 years developed persistant ulcerative skin lesions on her face and extremities. Granulomatous epitheloid tissue was found in skin biopsies, and later also in lung and tongue biopsies. No pathogens including mycobacteria were isolated. No improvement was seen on immunosuppression. Skin grafting was necessary in her lower extremity twice after partial destruction of the Achilles tendon by progressive ulceration. HSCT is planned in the near future.
Conclusion
RAG mutations may be associated with a combined immunodeficiency where uncontrolled granuloma formations of the skin, but not opportunistic infections, are leading clinical findings. It is unclear whether these disfiguring lesions are caused by pathogens such as mycobacteria which are difficult to culture or whether they represent reactive sarcoid-like granulomas as seen in other primary IDs, e.g. common variable immunodeficiency.
Specific Cytotoxicity and Clonal Expansion of TCR Vβ Subfamily T Cells Induced by PML-RARα Peptide.
Authors: Li Yangqiu , Tang Ji , Yang Lijian , Chen Shaohua , Zhou Yubing 
Published: April 2020

Abstract
The analysis of T cell receptor (TCR) Vβ repertoire is one of the sensitive methods to identify the clonal expansion T cells which response to tumor associated antigens. Understanding the clonality and restricted usage of TCR Vβ repertoire of expanded T-cells induced by PML-RARα peptide may be useful in helping design the new immunotherapeutic strategy specifically for acute promyelocytic leukemia (APL).The aim of the present study was to investigate the specific cytotoxicity and clonality of TCR Vβ repertoire in cord blood T cells induced by PML-RARα peptide (LSSCITQGKAIETQSSSSEE) in vitro. Cord blood mononuclear cells were amplified by IL-2, anti-CD3 and anti-CD28 antibody with different concentration (16.7μg /ml, 33.3μg /ml or 50μg /ml respectively) of synthetic PML-RARα peptide. The induced T cells were collected at different time points after culture (3, 6, 9, 10, 12 or 15 days). The expression and clonality of TCR Vβ subfamilies within induced T cells were analyzed by using RT-PCR and genescan technique. The cytotoxicity of induced T cells was detected by LDH release assay. The results showed that the best condition for T cells induction and amplication was at a concentration with 16.7μg /ml of PML-RARα peptide and at a culture duration with 10 to 15 day. TCR Vβ repertoire analysis showed that restricted expression and cloanl expansion of TCR Vβ subfamily cord blood T cells could be identified after induction by PML-RARα peptide. Clonal expanded T cells were found in Vβ13, Vβ14 and Vβ16 subfamlies respectively. The induced T cells were showed to have the specific cytotoxicity for NB4 cell line (effector cells: tagerted cells=20:1), the cytotoxicity rates were 49.65±6.7% (p&lt;0.05) at day 10th and 73.13±8.42% (p&lt;0.01) at day 15th after culture, which show statistical significance in compare to the control group (without PML-RARα induction). In conclusions, the PML-RARα peptide could induce the clonal expansion T cells from cord blood in vitro, which may have specific cytotoxicity for PML-RARα+ cells.
CD5+B Lymphocytes in the Bone Marrow of Immunic Hemocytopenia Patients and Its Clinical Significancy.
Authors: Shao Zonghong , Xing Limin , Fu Rong , Liu Hong , Shi Jun , Bai Jie , Tu Meifeng , Wang Huaquan , Cui Zhenzhu , Jia Hairong , Sun Juan , Yang Chongli 
Published: April 2020

Abstract
Background A subpopulation of B lymphocytes expressing CD5 antigen is involved in several autoimmune diseases through the release of autoantibodies. We investigate the quantitative of CD5+ B lymphocytes in the bone marrow of the patients with autoimmunic hemocytopenia and study the relationship between quantity of CD5+B cells and clinical or laboratorial parameters of these patients.Methods Quantities of CD5+B lymphocytes in the bone marrow(BM)of 14 patients with autoimmune hemolytic anemia(AIHA) or Evans syndrome, 22 immunorelated pancytopenia (IRP) patients and 10 normal controls were assayed by FACS. The correlation between their clinical or laboratory parameters and CD5+B lymphocytes was analyzed.
Results The qutantity of CD5+B lymphocytes of AIHA/Evans syndrome ((34.64±19.81)% or IRP((35.81±16.83)%) patients was significantly higher than that of normal controls((12.0±1.97)%)(p&lt;0.05). There was no significant difference between those of AIHA/Evans syndrome and IRP patients(p&gt;0.05). The quantity of CD5+B lymphocytes in the bone marrow of all cytopenic patients showed negative correlation with complement C3(r=−0.416, p&lt;0.05). In AIHA/Evans syndrome patients, the quantity of BM CD5+ B lymphocytes showed positive correlation with indirect bilirubin(IBIL) (r=1.00, p&lt;0.05); In Evans syndrome patients, the quantity of CD5+B lymphocytes in their bone marrow showed positive correlation with PAIgG (r=0.761, p&lt;0.05) and PAIgM(r=0.925, p&lt;0.05). The qutantity of CD5+B lymphocytes in the bone marrow of all cytopenic patients showed negative correlation with treatment response (correlation coefficient tau-b=−0.289, p&lt;0.05), but no relation with the yields of CFU-E, BFU-E,CFU-F and CFU-GM BM in vitro.
Conclusion BM CD5+B lymphocytes of the patients with autoimmunic hemocytopenias significantly increased and was related to the disease severity and clinical response. It was suggested that CD5+B lymphocytes might play an important role in the pathogenesis of autoimmunic hemocytopenia.
Gene Expression Profiling of Natural Killer Cell Subsets Isolated Using Novel MACS Methods.
Authors: Schmitz Jurgen 
Published: April 2020

Abstract
Natural Killer cells are now regarded as an important (Hokland, Mol Immunol 2005) but heterogenous (Cooper, Blood 2001) lymphocyte subset. Different Natural Killer cell subsets have different functions (Cooper, Blood 2001; Mavilio, PNAS 2005; Trotta, Blood 2005) and further research will be required to identify the role of those NK cell subsets in immune responses. We have developed novel methods, based on magnetic cell sorting, for purification of several NK cell subsets and have evaluated the separation performance of these kits using peripheral blood mononuclear cells from 10 donors. More than 1x105 CD56brightCD16− cells from 1x108 PBMC can be isolated with &gt;70% purity (&gt;85% CD56+CD16−), and more than 1x106 CD56dimCD16+ cells with &gt;94% purity. Additional subsets have been identified based on CD8 expression: &gt;2x106 CD8+ NK cells can be purified with &gt;90% purity.
Only limited data has previously been published on gene expression profiling of NK cell subsets using DNA microarrays (Koopman, J Exp Med 2003). We thus have used these MACS sorted NK cell subsets from 3 donors for an immune system specific microarray analysis (“PIQOR Immunology”). Usually gene expression profiling experiments are combined with flow cytometry characterization and functional tests. We used a novel amplification method to perform microarray analysis from mRNA isolated from as little as 1000 cells. Genes for surface molecules known to be differently expressed on NK cell subsets and immune effector molecules produced preferentially by single subsets served as control for the combination of amplification and microarray analysis (CD56, CD16, CD8, CD62L, CD122, Perforin, Granzyme B). Results of the analysis will be shown.