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Sample records for cell-derived il-2 signals

  1. Dendritic cell derived IL-2 inhibits survival of terminally mature cells via an autocrine signaling pathway.

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    Balachander, Akhila; Nabti, Sabrina; Sobota, Radoslaw M; Foo, Shihui; Zolezzi, Francesca; Lee, Bernett T K; Poidinger, Michael; Ricciardi-Castagnoli, Paola

    2015-05-01

    DCs are crucial for sensing pathogens and triggering immune response. Upon activation by pathogen-associated molecular pattern (PAMP) ligands, GM-CSF myeloid DCs (GM-DCs) secrete several cytokines, including IL-2. DC IL-2 has been shown to be important for innate and adaptive immune responses; however, IL-2 importance in DC physiology has never been demonstrated. Here, we show that autocrine IL-2 signaling is functional in murine GM-DCs in an early time window after PAMPs stimulation. IL-2 signaling selectively activates the JAK/STAT5 pathway by assembling holo-receptor complexes at the cell surface. Using the sensitivity of targeted mass spectrometry, we show conclusively that GM-DCs express CD122, the IL-2 receptor β-chain, at steady state. In myeloid DCs, this cytokine pathway inhibits survival of PAMP-matured GM-DCs which is crucial for maintaining immune tolerance and preventing autoimmunity. Our findings suggest that immune regulation by this novel autocrine signaling pathway can potentially be used in DC immunotherapy. PMID:25652593

  2. Curcumin blocks interleukin (IL)-2 signaling in T-lymphocytes by inhibiting IL-2 synthesis, CD25 expression, and IL-2 receptor signaling

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    Forward, Nicholas A.; Conrad, David M. [Department of Microbiology and Immunology, Dalhousie University, Halifax, Nova Scotia (Canada); Power Coombs, Melanie R.; Doucette, Carolyn D. [Department of Pathology, Dalhousie University, Halifax, Nova Scotia (Canada); Furlong, Suzanne J. [Department of Microbiology and Immunology, Dalhousie University, Halifax, Nova Scotia (Canada); Lin, Tong-Jun [Department of Microbiology and Immunology, Dalhousie University, Halifax, Nova Scotia (Canada); Department of Pediatrics, Dalhousie University, Halifax, Nova Scotia (Canada); Hoskin, David W., E-mail: d.w.hoskin@dal.ca [Department of Microbiology and Immunology, Dalhousie University, Halifax, Nova Scotia (Canada); Department of Pathology, Dalhousie University, Halifax, Nova Scotia (Canada); Department of Surgery, Dalhousie University, Halifax, Nova Scotia (Canada)

    2011-04-22

    Highlights: {yields} Curcumin inhibits CD4{sup +} T-lymphocyte proliferation. {yields} Curcumin inhibits interleukin-2 (IL-2) synthesis and CD25 expression by CD4{sup +} T-lymphocytes. {yields} Curcumin interferes with IL-2 receptor signaling by inhibiting JAK3 and STAT5 phosphorylation. {yields} IL-2-dependent regulatory T-lymphocyte function and Foxp3 expression is downregulated by curcumin. -- Abstract: Curcumin (diferulomethane) is the principal curcuminoid in the spice tumeric and a potent inhibitor of activation-induced T-lymphocyte proliferation; however, the molecular basis of this immunosuppressive effect has not been well studied. Here we show that micromolar concentrations of curcumin inhibited DNA synthesis by mouse CD4{sup +} T-lymphocytes, as well as interleukin-2 (IL-2) and CD25 ({alpha} chain of the high affinity IL-2 receptor) expression in response to antibody-mediated cross-linking of CD3 and CD28. Curcumin acted downstream of protein kinase C activation and intracellular Ca{sup 2+} release to inhibit I{kappa}B phosphorylation, which is required for nuclear translocation of the transcription factor NF{kappa}B. In addition, IL-2-dependent DNA synthesis by mouse CTLL-2 cells, but not constitutive CD25 expression, was impaired in the presence of curcumin, which demonstrated an inhibitory effect on IL-2 receptor (IL-2R) signaling. IL-2-induced phosphorylation of STAT5A and JAK3, but not JAK1, was diminished in the presence of curcumin, indicating inhibition of critical proximal events in IL-2R signaling. In line with the inhibitory action of curcumin on IL-2R signaling, pretreatment of CD4{sup +}CD25{sup +} regulatory T-cells with curcumin downregulated suppressor function, as well as forkhead box p3 (Foxp3) expression. We conclude that curcumin inhibits IL-2 signaling by reducing available IL-2 and high affinity IL-2R, as well as interfering with IL-2R signaling.

  3. Curcumin blocks interleukin (IL)-2 signaling in T-lymphocytes by inhibiting IL-2 synthesis, CD25 expression, and IL-2 receptor signaling

    International Nuclear Information System (INIS)

    Highlights: → Curcumin inhibits CD4+ T-lymphocyte proliferation. → Curcumin inhibits interleukin-2 (IL-2) synthesis and CD25 expression by CD4+ T-lymphocytes. → Curcumin interferes with IL-2 receptor signaling by inhibiting JAK3 and STAT5 phosphorylation. → IL-2-dependent regulatory T-lymphocyte function and Foxp3 expression is downregulated by curcumin. -- Abstract: Curcumin (diferulomethane) is the principal curcuminoid in the spice tumeric and a potent inhibitor of activation-induced T-lymphocyte proliferation; however, the molecular basis of this immunosuppressive effect has not been well studied. Here we show that micromolar concentrations of curcumin inhibited DNA synthesis by mouse CD4+ T-lymphocytes, as well as interleukin-2 (IL-2) and CD25 (α chain of the high affinity IL-2 receptor) expression in response to antibody-mediated cross-linking of CD3 and CD28. Curcumin acted downstream of protein kinase C activation and intracellular Ca2+ release to inhibit IκB phosphorylation, which is required for nuclear translocation of the transcription factor NFκB. In addition, IL-2-dependent DNA synthesis by mouse CTLL-2 cells, but not constitutive CD25 expression, was impaired in the presence of curcumin, which demonstrated an inhibitory effect on IL-2 receptor (IL-2R) signaling. IL-2-induced phosphorylation of STAT5A and JAK3, but not JAK1, was diminished in the presence of curcumin, indicating inhibition of critical proximal events in IL-2R signaling. In line with the inhibitory action of curcumin on IL-2R signaling, pretreatment of CD4+CD25+ regulatory T-cells with curcumin downregulated suppressor function, as well as forkhead box p3 (Foxp3) expression. We conclude that curcumin inhibits IL-2 signaling by reducing available IL-2 and high affinity IL-2R, as well as interfering with IL-2R signaling.

  4. 4-1BB Signaling Enhances Primary and Secondary Population Expansion of CD8+ T Cells by Maximizing Autocrine IL-2/IL-2 Receptor Signaling.

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    Ho S Oh

    Full Text Available 4-1BB (CD137, a member of the tumor necrosis factor receptor superfamily (TNFRSF, is primarily expressed on activated T cells and is known to enhance proliferation of T cells, prevent activation-induced cell death, and promote memory formation of CD8+ T cells. In particular, it is well acknowledged that 4-1BB triggering preferentially enhances the expansion of CD8+ T cells rather than CD4+ T cells, but the underlying mechanism remains unclear. Here we found that 4-1BB triggering markedly increased IL-2Rα (CD25 and IL-2 expressions of CD8+ T cells but minimally for CD4+ T cells. Proliferation of CD8+ T cells was moderately enhanced by direct 4-1BB triggering in the absence of signaling through IL-2Rα/IL-2 interactions, but further promoted in the presence of IL-2Rα/IL-2 interactions. Among the TNFRSF members including OX40, GITR, CD30, and CD27, 4-1BB was superior in the ability to induce IL-2Rα expression on CD8+ T cells. When the primary and secondary expansions of CD8+ T cells in vivo were examined by adoptively transferring OVA-specific CD8+ T cells along with the treatment with agonistic anti-4-1BB and/or antagonistic anti-CD25 F(ab'2 mAb, 4-1BB triggering enhanced both primary and secondary expansion of CD8+ T cells in vivo, and the 4-1BB effects were moderately suppressed in primary expansion while completely abolished in secondary expansion of OVA-specific CD8+ T cells by blocking IL-2Rα. These results suggest that 4-1BB-mediated increases of IL-2Rα and IL-2 prolong the effects of transient TCR- and 4-1BB-mediated signaling in CD8+ T cells, and that 4-1BB triggering preferentially enhances the expansion of CD8+ T cells through the amplification of autocrine IL-2/IL-2R signaling loop.

  5. 4-1BB Signaling Enhances Primary and Secondary Population Expansion of CD8+ T Cells by Maximizing Autocrine IL-2/IL-2 Receptor Signaling.

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    Oh, Ho S; Choi, Beom K; Kim, Young H; Lee, Don G; Hwang, Sunhee; Lee, Myoung J; Park, Sang H; Bae, Yong-Soo; Kwon, Byoung S

    2015-01-01

    4-1BB (CD137), a member of the tumor necrosis factor receptor superfamily (TNFRSF), is primarily expressed on activated T cells and is known to enhance proliferation of T cells, prevent activation-induced cell death, and promote memory formation of CD8+ T cells. In particular, it is well acknowledged that 4-1BB triggering preferentially enhances the expansion of CD8+ T cells rather than CD4+ T cells, but the underlying mechanism remains unclear. Here we found that 4-1BB triggering markedly increased IL-2Rα (CD25) and IL-2 expressions of CD8+ T cells but minimally for CD4+ T cells. Proliferation of CD8+ T cells was moderately enhanced by direct 4-1BB triggering in the absence of signaling through IL-2Rα/IL-2 interactions, but further promoted in the presence of IL-2Rα/IL-2 interactions. Among the TNFRSF members including OX40, GITR, CD30, and CD27, 4-1BB was superior in the ability to induce IL-2Rα expression on CD8+ T cells. When the primary and secondary expansions of CD8+ T cells in vivo were examined by adoptively transferring OVA-specific CD8+ T cells along with the treatment with agonistic anti-4-1BB and/or antagonistic anti-CD25 F(ab')2 mAb, 4-1BB triggering enhanced both primary and secondary expansion of CD8+ T cells in vivo, and the 4-1BB effects were moderately suppressed in primary expansion while completely abolished in secondary expansion of OVA-specific CD8+ T cells by blocking IL-2Rα. These results suggest that 4-1BB-mediated increases of IL-2Rα and IL-2 prolong the effects of transient TCR- and 4-1BB-mediated signaling in CD8+ T cells, and that 4-1BB triggering preferentially enhances the expansion of CD8+ T cells through the amplification of autocrine IL-2/IL-2R signaling loop. PMID:25962156

  6. Manipulating Memory CD8 T Cell Numbers by Timed Enhancement of IL-2 Signals.

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    Kim, Marie T; Kurup, Samarchith P; Starbeck-Miller, Gabriel R; Harty, John T

    2016-09-01

    As a result of the growing burden of tumors and chronic infections, manipulating CD8 T cell responses for clinical use has become an important goal for immunologists. In this article, we show that dendritic cell (DC) immunization coupled with relatively early (days 1-3) or late (days 4-6) administration of enhanced IL-2 signals increase peak effector CD8 T cell numbers, but only early IL-2 signals enhance memory numbers. IL-2 signals delivered at relatively late time points drive terminal differentiation and marked Bim-mediated contraction and do not increase memory T cell numbers. In contrast, early IL-2 signals induce effector cell metabolic profiles that are more conducive to memory formation. Of note, downregulation of CD80 and CD86 was observed on DCs in vivo following early IL-2 treatment. Mechanistically, early IL-2 treatment enhanced CTLA-4 expression on regulatory T cells, and CTLA-4 blockade alongside IL-2 treatment in vivo prevented the decrease in CD80 and CD86, supporting a cell-extrinsic role for CTLA-4 in downregulating B7 ligand expression on DCs. Finally, DC immunization followed by early IL-2 treatment and anti-CTLA-4 blockade resulted in lower memory CD8 T cell numbers compared with the DC+early IL-2 treatment group. These data suggest that curtailed signaling through the B7-CD28 costimulatory axis during CD8 T cell activation limits terminal differentiation and preserves memory CD8 T cell formation; thus, it should be considered in future T cell-vaccination strategies. PMID:27439516

  7. Activated human T cells secrete exosomes that participate in IL-2 mediated immune response signaling.

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    Jessica Wahlgren

    Full Text Available It has previously been shown that nano-meter sized vesicles (30-100 nm, exosomes, secreted by antigen presenting cells can induce T cell responses thus showing the potential of exosomes to be used as immunological tools. Additionally, activated CD3⁺ T cells can secrete exosomes that have the ability to modulate different immunological responses. Here, we investigated what effects exosomes originating from activated CD3⁺ T cells have on resting CD3⁺ T cells by studying T cell proliferation, cytokine production and by performing T cell and exosome phenotype characterization. Human exosomes were generated in vitro following CD3⁺ T cell stimulation with anti-CD28, anti-CD3 and IL-2. Our results show that exosomes purified from stimulated CD3⁺ T cells together with IL-2 were able to generate proliferation in autologous resting CD3⁺ T cells. The CD3⁺ T cells stimulated with exosomes together with IL-2 had a higher proportion of CD8⁺ T cells and had a different cytokine profile compared to controls. These results indicate that activated CD3⁺ T cells communicate with resting autologous T cells via exosomes.

  8. Resolving Early Signaling Events in T-Cell Activation Leading to IL-2 and FOXP3 Transcription

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    Jeffrey P. Perley

    2014-11-01

    Full Text Available Signal intensity and feedback regulation are known to be major factors in the signaling events stemming from the T-cell receptor (TCR and its various coreceptors, but the exact nature of these relationships remains in question. We present a mathematical model of the complex signaling network involved in T-cell activation with cross-talk between the Erk, calcium, PKC and mTOR signaling pathways. The model parameters are adjusted to fit new and published data on TCR trafficking, Zap70, calcium, Erk and Isignaling. The regulation of the early signaling events by phosphatases, CD45 and SHP1, and the TCR dynamics are critical to determining the behavior of the model. Additional model corroboration is provided through quantitative and qualitative agreement with experimental data collected under different stimulating and knockout conditions. The resulting model is analyzed to investigate how signal intensity and feedback regulation affect TCR- and coreceptor-mediated signal transduction and their downstream transcriptional profiles to predict the outcome for a variety of stimulatory and knockdown experiments. Analysis of the model shows that: (1 SHP1 negative feedback is necessary for preventing hyperactivity in TCR signaling; (2 CD45 is required for TCR signaling, but also partially suppresses it at high expression levels; and (3 elevated FOXP3 and reduced IL-2 signaling, an expression profile often associated with T regulatory cells (Tregs, is observed when the system is subjected to weak TCR and CD28 costimulation or a severe reduction in CD45 activity.

  9. Malignant transformation of CD4+ T lymphocytes mediated by oncogenic kinase NPM/ALK recapitulates IL-2-induced cell signaling and gene expression reprogramming.

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    Marzec, Michal; Halasa, Krzysztof; Liu, Xiaobin; Wang, Hong Y; Cheng, Mangeng; Baldwin, Donald; Tobias, John W; Schuster, Stephen J; Woetmann, Anders; Zhang, Qian; Turner, Suzanne D; Ødum, Niels; Wasik, Mariusz A

    2013-12-15

    Anaplastic lymphoma kinase (ALK), physiologically expressed only by nervous system cells, displays a remarkable capacity to transform CD4(+) T lymphocytes and other types of nonneural cells. In this study, we report that activity of nucleophosmin (NPM)/ALK chimeric protein, the dominant form of ALK expressed in T cell lymphomas (TCLs), closely resembles cell activation induced by IL-2, the key cytokine supporting growth and survival of normal CD4(+) T lymphocytes. Direct comparison of gene expression by ALK(+) TCL cells treated with an ALK inhibitor and IL-2-dependent ALK(-) TCL cells stimulated with the cytokine revealed a very similar, albeit inverse, gene-regulation pattern. Depending on the analysis method, up to 67% of the affected genes were modulated in common by NPM/ALK and IL-2. Based on the gene expression patterns, Jak/STAT- and IL-2-signaling pathways topped the list of pathways identified as affected by both IL-2 and NPM/ALK. The expression dependence on NPM/ALK and IL-2 of the five selected genes-CD25 (IL-2Rα), Egr-1, Fosl-1, SOCS3, and Irf-4-was confirmed at the protein level. In both ALK(+) TCL and IL-2-stimulated ALK(-) TCL cells, CD25, SOCS3, and Irf-4 genes were activated predominantly by the STAT5 and STAT3 transcription factors, whereas transcription of Egr-1 and Fosl-1 was induced by the MEK-ERK pathway. Finally, we found that Egr-1, a protein not associated previously with either IL-2 or ALK, contributes to the cell proliferation. These findings indicate that NPM/ALK transforms the target CD4(+) T lymphocytes, at least in part, by using the pre-existing, IL-2-dependent signaling pathways.

  10. Malignant transformation of CD4+ T lymphocytes mediated by oncogenic kinase NPM/ALK recapitulates IL-2-induced cell signaling and gene expression reprogramming

    DEFF Research Database (Denmark)

    Marzec, Michal; Halasa, Krzysztof; Liu, Xiaobin;

    2013-01-01

    Anaplastic lymphoma kinase (ALK), physiologically expressed only by nervous system cells, displays a remarkable capacity to transform CD4(+) T lymphocytes and other types of nonneural cells. In this study, we report that activity of nucleophosmin (NPM)/ALK chimeric protein, the dominant form of ALK...... with the cytokine revealed a very similar, albeit inverse, gene-regulation pattern. Depending on the analysis method, up to 67% of the affected genes were modulated in common by NPM/ALK and IL-2. Based on the gene expression patterns, Jak/STAT- and IL-2-signaling pathways topped the list of pathways identified...... as affected by both IL-2 and NPM/ALK. The expression dependence on NPM/ALK and IL-2 of the five selected genes-CD25 (IL-2Rα), Egr-1, Fosl-1, SOCS3, and Irf-4-was confirmed at the protein level. In both ALK(+) TCL and IL-2-stimulated ALK(-) TCL cells, CD25, SOCS3, and Irf-4 genes were activated predominantly...

  11. MHC class II molecules deliver costimulatory signals in human T cells through a functional linkage with IL-2-receptors

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    Odum, Niels; Kanner, S B; Ledbetter, J A;

    1993-01-01

    a regulatory function in T cell activation. Here, we show that cross-linking HLA-DR and -DP but not -DQ molecules by immobilized mAb enhanced proliferative T cell responses to IL-2. In contrast, class II stimulation had no effect on IL-4-induced proliferation. The costimulatory effect was most......Ab induced tyrosine phosphorylation of specific substrates including PLC-gamma 1. Combined stimulation of IL-2R and class II molecules had an additive effect on tyrosine phosphorylation. Pretreatment of T cells with a protein tyrosine kinase inhibitor, herbimycin A, inhibited IL-2 and class II...

  12. Decreased SAP Expression in T Cells from Patients with Systemic Lupus Erythematosus Contributes to Early Signaling Abnormalities and Reduced IL-2 Production.

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    Karampetsou, Maria P; Comte, Denis; Kis-Toth, Katalin; Terhorst, Cox; Kyttaris, Vasileios C; Tsokos, George C

    2016-06-15

    T cells from patients with systemic lupus erythematosus (SLE) display a number of abnormalities, including increased early signaling events following engagement of the TCR. Signaling lymphocytic activation molecule family cell surface receptors and the X-chromosome-defined signaling lymphocytic activation molecule-associated protein (SAP) adaptor are important in the development of several immunocyte lineages and modulating the immune response. We present evidence that SAP protein levels are decreased in T cells and in their main subsets isolated from 32 women and three men with SLE, independent of disease activity. In SLE T cells, SAP protein is also subject to increased degradation by caspase-3. Forced expression of SAP in SLE T cells normalized IL-2 production, calcium (Ca(2+)) responses, and tyrosine phosphorylation of a number of proteins. Exposure of normal T cells to SLE serum IgG, known to contain anti-CD3/TCR Abs, resulted in SAP downregulation. We conclude that SLE T cells display reduced levels of the adaptor protein SAP, probably as a result of continuous T cell activation and degradation by caspase-3. Restoration of SAP levels in SLE T cells corrects the overexcitable lupus T cell phenotype.

  13. Tip cell-derived RTK signaling initiates cell movements in the Drosophila stomatogastric nervous system anlage.

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    González-Gaitán, M; Jäckle, H

    2000-10-01

    The stomatogastric nervous system (SNS) of Drosophila is a simply organized neural circuitry that innervates the anterior enteric system. Unlike the central and the peripheral nervous systems, the SNS derives from a compact epithelial anlage in which three invagination centers, each giving rise to an invagination fold headed by a tip cell, are generated. Tip cell selection involves lateral inhibition, a process in which Wingless (Wg) activity adjusts the range of Notch signaling. Here we show that RTK signaling mediated by the Drosophila homolog of the epidermal growth factor receptor, DER, plays a key role in two consecutive steps during early SNS development. Like Wg, DER signaling participates in adjusting the range of Notch-dependent lateral inhibition during tip cell selection. Subsequently, tip cells secrete the DER ligand Spitz and trigger local RTK signaling, which initiates morphogenetic movements resulting in the tip cell-directed invaginations within the SNS anlage.

  14. A new system for profiling drug-induced calcium signal perturbation in human embryonic stem cell-derived cardiomyocytes.

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    Lewis, Kimberley J; Silvester, Nicole C; Barberini-Jammaers, Steven; Mason, Sammy A; Marsh, Sarah A; Lipka, Magdalena; George, Christopher H

    2015-03-01

    The emergence of human stem cell-derived cardiomyocyte (hSCCM)-based assays in the cardiovascular (CV) drug discovery sphere requires the development of improved systems for interrogating the rich information that these cell models have the potential to yield. We developed a new analytical framework termed SALVO (synchronization, amplitude, length, and variability of oscillation) to profile the amplitude and temporal patterning of intra- and intercellular calcium signals in hSCCM. SALVO quantified drug-induced perturbations in the calcium signaling "fingerprint" in spontaneously contractile hSCCM. Multiparametric SALVO outputs were integrated into a single index of in vitro cytotoxicity that confirmed the rank order of perturbation as astemizole > thioridazine > cisapride > flecainide > valdecoxib > sotalol > nadolol ≈ control. This rank order of drug-induced Ca(2+) signal disruption is in close agreement with the known arrhythmogenic liabilities of these compounds in humans. Validation of the system using a second set of compounds and hierarchical cluster analysis demonstrated the utility of SALVO to discriminate drugs based on their mechanisms of action. We discuss the utility of this new mechanistically agnostic system for the evaluation of in vitro drug cytotoxicity in hSCCM syncytia and the potential placement of SALVO in the early stage drug screening framework. PMID:25367900

  15. Radio-sensitivities and angiogenic signaling pathways of irradiated normal endothelial cells derived from diverse human organs

    International Nuclear Information System (INIS)

    The purpose of the present investigation was to study the effects of ionizing radiation on endothelial cells derived from diverse normal tissues. We first compared the effects of radiation on clonogenic survival and tube formation of endothelial cells, and then investigated the molecular signaling pathways involved in endothelial cell survival and angiogenesis. Among the different endothelial cells studied, human hepatic sinusoidal endothelial cells (HHSECs) were the most radio-resistant and human dermal microvascular endothelial cells were the most radio-sensitive. The radio-resistance of HHSECs was related to adenosine monophosphate-activated protein kinase and p38 mitogen-activated protein kinase-mediated expression of MMP-2 and vascular endothelial growth factor receptor-2 (VEGFR-2), whereas the increased radio-sensitivity of HDMECs was related to extracellular signal-regulated kina0se-mediated generation of angiostatin. These observations demonstrate that there are distinct differences in the radiation responses of normal endothelial cells obtained from diverse organs, which may provide important clues for protection of normal tissue from radiation exposure. (author)

  16. Stroma cell-derived factor-1α signaling enhances calcium transients and beating frequency in rat neonatal cardiomyocytes.

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    Ielham Hadad

    Full Text Available Stroma cell-derived factor-1α (SDF-1α is a cardioprotective chemokine, acting through its G-protein coupled receptor CXCR4. In experimental acute myocardial infarction, administration of SDF-1α induces an early improvement of systolic function which is difficult to explain solely by an anti-apoptotic and angiogenic effect. We wondered whether SDF-1α signaling might have direct effects on calcium transients and beating frequency.Primary rat neonatal cardiomyocytes were culture-expanded and characterized by immunofluorescence staining. Calcium sparks were studied by fluorescence microscopy after calcium loading with the Fluo-4 acetoxymethyl ester sensor. The cardiomyocyte enriched cellular suspension expressed troponin I and CXCR4 but was vimentin negative. Addition of SDF-1α in the medium increased cytoplasmic calcium release. The calcium response was completely abolished by using a neutralizing anti-CXCR4 antibody and partially suppressed and delayed by preincubation with an inositol triphosphate receptor (IP3R blocker, but not with a ryanodine receptor (RyR antagonist. Calcium fluxes induced by caffeine, a RyR agonist, were decreased by an IP3R blocker. Treatment with forskolin or SDF-1α increased cardiomyocyte beating frequency and their effects were additive. In vivo, treatment with SDF-1α increased left ventricular dP/dtmax.These results suggest that in rat neonatal cardiomyocytes, the SDF-1α/CXCR4 signaling increases calcium transients in an IP3-gated fashion leading to a positive chronotropic and inotropic effect.

  17. TOM1L Is Involved in a Novel Signaling Pathway Important for the IL-2 Production in Jurkat T Cells Stimulated by CD3/CD28 CoLigation

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    Ahmed Elmarghani

    2009-01-01

    Full Text Available TOM1L (target of Myb-1 Like was identified as a binding partner for the full length and catalytically-active Lck in a yeast 2-hybrid screening assay. Here we show that in Jurkat T cells stimulated by CD3/CD28 coligation where the expression of TOM1L is reduced by lenti virus mediated-siRNA results in a dramatically lower IL-2 production. The production of IL-2 in siRNA treated cells stimulated with PMA/ionomycin was not affected indicating an involvement of TOM1L in a pathway proximal of TCR and CD28. The coexpression of Fyn with TOM1L increased the level of the phosphorylated form of Fyn indicating that TOM1L has the ability to activate Fyn. The ability of TOM1L to activate Fyn was further shown in a kinase assay using angiotensin II as a substrate. By confocal microscopy, we show that the expression of TOM1L in non-treated HeLa and SK-N-SH cells colocalizes with the mitochondrial membrane but not with lysosomal compartments or the trans-Golgi network. Furthermore, we show that the over-expression of TOM1L in Jurkat cells causes an increase of the STAT3 expression . Based on our results, we here propose that TOM1L is involved in a novel signaling pathway that is important for the IL-2 production in T cells.

  18. Melanoma cell-derived exosomes promote epithelial-mesenchymal transition in primary melanocytes through paracrine/autocrine signaling in the tumor microenvironment.

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    Xiao, Deyi; Barry, Samantha; Kmetz, Daniel; Egger, Michael; Pan, Jianmin; Rai, Shesh N; Qu, Jifu; McMasters, Kelly M; Hao, Hongying

    2016-07-01

    The tumor microenvironment is abundant with exosomes that are secreted by the cancer cells themselves. Exosomes are nanosized, organelle-like membranous structures that are increasingly being recognized as major contributors in the progression of malignant neoplasms. A critical element in melanoma progression is its propensity to metastasize, but little is known about how melanoma cell-derived exosomes modulate the microenvironment to optimize conditions for tumor progression and metastasis. Here, we provide evidence that melanoma cell-derived exosomes promote phenotype switching in primary melanocytes through paracrine/autocrine signaling. We found that the mitogen-activated protein kinase (MAPK) signaling pathway was activated during the exosome-mediated epithelial-to-mesenchymal transition (EMT)-resembling process, which promotes metastasis. Let-7i, an miRNA modulator of EMT, was also involved in this process. We further defined two other miRNA modulators of EMT (miR-191 and let-7a) in serum exosomes for differentiating stage I melanoma patients from non-melanoma subjects. These results provide the first strong molecular evidence that melanoma cell-derived exosomes promote the EMT-resembling process in the tumor microenvironment. Thus, novel strategies targeting EMT and modulating the tumor microenvironment may emerge as important approaches for the treatment of metastatic melanoma. PMID:27063098

  19. Interleukin-2 (IL-2) dependent expression of biologically relevant IL-2 receptors: uncoupling of anti-T3 induced receptor expression with cyclosporin

    International Nuclear Information System (INIS)

    Human peripheral blood T cell expression of IL-2 receptors (IL-2R), detected by both immunocytofluorometry and 125I-IL-2 binding, was studied using lymphocytes stimulated with monoclonal anti-T3 antibodies (Leu-4, OKT3). Lymphocytes, isolated from healthy individuals, were prescreened and classified as Leu-4 responders or non-responders according to 72 h 3H-thymidine incorporation experiments. Leu-4 non-responder lymphocytes, though capable of normal IL-2R expression and IL-2 secretion when cultured with OKT3 (IgG2a), expressed little to no IL-2R nor secreted IL-2 when stimulated with Leu-4 (IgG1). In addition, the amount of IL-2 secreted by Leu-4 stimulated, Leu-4 responder cells, was one-third- to one-fifth of that detected when OKT3 was used as the stimulant. The addition of recombinant IL-2 (rIL-2) to a Leu-4 stimulated, Leu-4 non-responder lymphocyte culture, resulted in the expression of IL-2R and cellular proliferation, indicating that IL-2 upregulated its biologically relevant receptor. As expected, cyclosporin-A (CSA) inhibited the secretion of IL-2 and subsequent proliferation of Leu-4 stimulated, Leu-4 responder cells. Unexpectedly, however, the expression of IL-2R was also blocked. Exogenous rIL-2 partially reversed the effect of CSA on IL-2R expression and proliferation. The results indicate that IL-2 may provide an additional, required signal for optimal IL-2R expression

  20. Community effect triggers terminal differentiation of myogenic cells derived from muscle satellite cells by quenching Smad signaling

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    Yanagisawa, Michiko [Department of Regenerative Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, 35 Gengo, Morioka, Oobu, Aichi 474-8522 (Japan); Aging Research, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi 466-8550 (Japan); Mukai, Atsushi; Shiomi, Kosuke [Department of Regenerative Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, 35 Gengo, Morioka, Oobu, Aichi 474-8522 (Japan); Song, Si-Yong [Institute of Neuroscience, Faculty of Pharmaceutical Sciences at Kagawa, Tokushima Bunri University, 1314-1 Shido, Sanuki-shi, Kagawa 769-2193 (Japan); Hashimoto, Naohiro, E-mail: nao@ncgg.go.jp [Department of Regenerative Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, 35 Gengo, Morioka, Oobu, Aichi 474-8522 (Japan)

    2011-01-15

    A high concentration of bone morphogenetic proteins (BMPs) stimulates myogenic progenitor cells to undergo heterotopic osteogenic differentiation. However, the physiological role of the Smad signaling pathway during terminal muscle differentiation has not been resolved. We report here that Smad1/5/8 was phosphorylated and activated in undifferentiated growing mouse myogenic progenitor Ric10 cells without exposure to any exogenous BMPs. The amount of phosphorylated Smad1/5/8 was severely reduced during precocious myogenic differentiation under the high cell density culture condition even in growth medium supplemented with a high concentration of serum. Inhibition of the Smad signaling pathway by dorsomorphin, an inhibitor of Smad activation, or noggin, a specific antagonist of BMP, induced precocious terminal differentiation of myogenic progenitor cells in a cell density-dependent fashion even in growth medium. In addition, Smad1/5/8 was transiently activated in proliferating myogenic progenitor cells during muscle regeneration in rats. The present results indicate that the Smad signaling pathway is involved in a critical switch between growth and differentiation of myogenic progenitor cells both in vitro and in vivo. Furthermore, precocious cell density-dependent myogenic differentiation suggests that a community effect triggers the terminal muscle differentiation of myogenic cells by quenching the Smad signaling.

  1. Community effect triggers terminal differentiation of myogenic cells derived from muscle satellite cells by quenching Smad signaling

    International Nuclear Information System (INIS)

    A high concentration of bone morphogenetic proteins (BMPs) stimulates myogenic progenitor cells to undergo heterotopic osteogenic differentiation. However, the physiological role of the Smad signaling pathway during terminal muscle differentiation has not been resolved. We report here that Smad1/5/8 was phosphorylated and activated in undifferentiated growing mouse myogenic progenitor Ric10 cells without exposure to any exogenous BMPs. The amount of phosphorylated Smad1/5/8 was severely reduced during precocious myogenic differentiation under the high cell density culture condition even in growth medium supplemented with a high concentration of serum. Inhibition of the Smad signaling pathway by dorsomorphin, an inhibitor of Smad activation, or noggin, a specific antagonist of BMP, induced precocious terminal differentiation of myogenic progenitor cells in a cell density-dependent fashion even in growth medium. In addition, Smad1/5/8 was transiently activated in proliferating myogenic progenitor cells during muscle regeneration in rats. The present results indicate that the Smad signaling pathway is involved in a critical switch between growth and differentiation of myogenic progenitor cells both in vitro and in vivo. Furthermore, precocious cell density-dependent myogenic differentiation suggests that a community effect triggers the terminal muscle differentiation of myogenic cells by quenching the Smad signaling.

  2. Cell proliferation potency is independent of FGF4 signaling in trophoblast stem cells derived from androgenetic embryos

    OpenAIRE

    Ogawa, Hidehiko; TAKYU, Ryuichi; MORIMOTO, Hiromu; TOEI, Shuntaro; SAKON, Hiroshi; GOTO, Shiori; MORIYA, SHOTA; Kono, Tomohiro

    2015-01-01

    We previously established trophoblast stem cells from mouse androgenetic embryos (AGTS cells). In this study, to further characterize AGTS cells, we compared cell proliferation activity between trophoblast stem (TS) cells and AGTS cells under fibroblast growth factor 4 (FGF4) signaling. TS cells continued to proliferate and maintained mitotic cell division in the presence of FGF4. After FGF4 deprivation, the cell proliferation stopped, the rate of M-phase cells decreased, and trophoblast gian...

  3. Biological significance of soluble IL-2 receptor

    Directory of Open Access Journals (Sweden)

    Calogero Caruso

    1993-01-01

    Full Text Available A NUMBER of receptors for growth factors and differentiation antigens have been found to be secreted or released by cells. Following mononuclear cell (MNC activation and interleukin-2 receptor (IL-2R expression, a soluble form of the Alpha;-chain of IL-2R (sIL-2R is released. The sIL-2R has been shown to be present in the culture supernatants of activated MNCs as well as in normal sera and, in higher amounts, in sera from subjects affected by several diseases including neoplastic, infectious and autoimmune ones, and in sera from transplanted patients suffering allograft rejection. The blood sIL-2R levels depend on the number of producing cells and the number of molecules per cell, so that sIL-2R blood values may represent an index of the number and the functional state of producing cells, both normal and neoplastic. Thus, monitoring of the immune system, mostly T-cells and haematological malignancies might be targets for the measurement of sIL-2R. Since many conditions may influence sIL-2R production, little diagnostic use may result from these measurements. However, since blood sIL-2R levels may correlate with disease progression and/or response to therapy, their measurement may be a useful index of activity and extent of disease. The precise biological role of the soluble form of the IL-2R is still a matter of debate. However, we know that increased sIL-2R levels may be observed in association with several immunological abnormalities and that sIL-2R is able to bind IL-2. It is conceivable then that in these conditions the excess sIL-2R released in vivo by activated lymphoid cells or by neoplastic cells may somehow regulate IL-2-dependent processes. On the other hand, it cannot exclude that sIL-2R is a by-product without biological significance. Finally, it is puzzling that in many conditions in which an increase of blood sIL-2R values has been observed, MNCs display a decreased in vitro capacity to produce sIL-2R. These seemingly contrasting

  4. Gene Expression Profile Reveals Abnormalities of Multiple Signaling Pathways in Mesenchymal Stem Cell Derived from Patients with Systemic Lupus Erythematosus

    Directory of Open Access Journals (Sweden)

    Yu Tang

    2012-01-01

    Full Text Available We aimed to compare bone-marrow-derived mesenchymal stem cells (BMMSCs between systemic lupus erythematosus (SLE and normal controls by means of cDNA microarray, immunohistochemistry, immunofluorescence, and immunoblotting. Our results showed there were a total of 1, 905 genes which were differentially expressed by BMMSCs derived from SLE patients, of which, 652 genes were upregulated and 1, 253 were downregulated. Gene ontology (GO analysis showed that the majority of these genes were related to cell cycle and protein binding. Pathway analysis exhibited that differentially regulated signal pathways involved actin cytoskeleton, focal adhesion, tight junction, and TGF-β pathway. The high protein level of BMP-5 and low expression of Id-1 indicated that there might be dysregulation in BMP/TGF-β signaling pathway. The expression of Id-1 in SLE BMMSCs was reversely correlated with serum TNF-α levels. The protein level of cyclin E decreased in the cell cycling regulation pathway. Moreover, the MAPK signaling pathway was activated in BMMSCs from SLE patients via phosphorylation of ERK1/2 and SAPK/JNK. The actin distribution pattern of BMMSCs from SLE patients was also found disordered. Our results suggested that there were distinguished differences of BMMSCs between SLE patients and normal controls.

  5. IL-1β-induced, matrix metalloproteinase-3-regulated proliferation of embryonic stem cell-derived odontoblastic cells is mediated by the Wnt5 signaling pathway

    International Nuclear Information System (INIS)

    We previously established a method for differentiating induced pluripotent stem cells and embryonic stem (ES) cells into α2 integrin-positive odontoblast-like cells. We also reported that interleukin (IL)-1β induces matrix metalloproteinase (MMP)-3-regulated cell proliferation and suppresses apoptosis in these cells, suggesting that MMP-3 plays a potentially unique physiological role in the regeneration of odontoblast-like cells. Here, we examined whether up-regulation of MMP-3 activity by IL-1β was mediated by Wnt signaling and led to increased proliferation of odontoblast-like cells. IL-1β increased mRNA and protein levels of Wnt5a, Wnt5b and the Wnt receptor Lrp5. Exogenous Wnt5a and Wnt5b were found to increase MMP-3 mRNA, protein and activity, and interestingly the rate of proliferation in these cells. Treatment with siRNAs against Wnt5a, Wnt5b and Lrp5 suppressed the IL-1β-induced increase in MMP-3 expression and suppressed cell proliferation, an effect rescued by application of exogenous Wnt5. These results demonstrate the sequential involvement of Wnt5, Lrp5 and MMP-3 in effecting IL-1β-induced proliferation of ES cell-derived odontoblast-like cells. - Highlights: • IL-1β induces Wnt5, Lrp5/Fzd9 and MMP-3 in ES cell-derived odontoblast-like cells. • IL-1β-induced Wnt5 expression results in increased cell proliferation. • Exogenous Wnt5 increases MMP-3 activity and cell proliferation. • Exogenous Wnt5 rescues IL-1β-driven proliferation with anti-Wnt5 siRNA suppression. • IL-1β-induced cell proliferation involves Wnt5, Lrp5, and MMP-3 sequentially

  6. IL-1β-induced, matrix metalloproteinase-3-regulated proliferation of embryonic stem cell-derived odontoblastic cells is mediated by the Wnt5 signaling pathway

    Energy Technology Data Exchange (ETDEWEB)

    Ozeki, Nobuaki; Hase, Naoko; Hiyama, Taiki; Yamaguchi, Hideyuki; Kawai, Rie [Department of Endodontics, School of Dentistry, Aichi Gakuin University, Nagoya, Aichi 464-8651 (Japan); Kondo, Ayami [Department of Medicinal Biochemistry, School of Pharmacy, Aichi Gakuin University, Nagoya 464-8650 (Japan); Nakata, Kazuhiko [Department of Endodontics, School of Dentistry, Aichi Gakuin University, Nagoya, Aichi 464-8651 (Japan); Mogi, Makio, E-mail: makio@dpc.agu.ac.jp [Department of Medicinal Biochemistry, School of Pharmacy, Aichi Gakuin University, Nagoya 464-8650 (Japan)

    2014-10-15

    We previously established a method for differentiating induced pluripotent stem cells and embryonic stem (ES) cells into α2 integrin-positive odontoblast-like cells. We also reported that interleukin (IL)-1β induces matrix metalloproteinase (MMP)-3-regulated cell proliferation and suppresses apoptosis in these cells, suggesting that MMP-3 plays a potentially unique physiological role in the regeneration of odontoblast-like cells. Here, we examined whether up-regulation of MMP-3 activity by IL-1β was mediated by Wnt signaling and led to increased proliferation of odontoblast-like cells. IL-1β increased mRNA and protein levels of Wnt5a, Wnt5b and the Wnt receptor Lrp5. Exogenous Wnt5a and Wnt5b were found to increase MMP-3 mRNA, protein and activity, and interestingly the rate of proliferation in these cells. Treatment with siRNAs against Wnt5a, Wnt5b and Lrp5 suppressed the IL-1β-induced increase in MMP-3 expression and suppressed cell proliferation, an effect rescued by application of exogenous Wnt5. These results demonstrate the sequential involvement of Wnt5, Lrp5 and MMP-3 in effecting IL-1β-induced proliferation of ES cell-derived odontoblast-like cells. - Highlights: • IL-1β induces Wnt5, Lrp5/Fzd9 and MMP-3 in ES cell-derived odontoblast-like cells. • IL-1β-induced Wnt5 expression results in increased cell proliferation. • Exogenous Wnt5 increases MMP-3 activity and cell proliferation. • Exogenous Wnt5 rescues IL-1β-driven proliferation with anti-Wnt5 siRNA suppression. • IL-1β-induced cell proliferation involves Wnt5, Lrp5, and MMP-3 sequentially.

  7. Th cells promote CTL survival and memory via acquired pMHC-I and endogenous IL-2 and CD40L signaling and by modulating apoptosis-controlling pathways.

    Directory of Open Access Journals (Sweden)

    Channakeshava Sokke Umeshappa

    Full Text Available Involvement of CD4(+ helper T (Th cells is crucial for CD8(+ cytotoxic T lymphocyte (CTL-mediated immunity. However, CD4(+ Th's signals that govern CTL survival and functional memory are still not completely understood. In this study, we assessed the role of CD4(+ Th cells with acquired antigen-presenting machineries in determining CTL fates. We utilized an adoptive co-transfer into CD4(+ T cell-sufficient or -deficient mice of OTI CTLs and OTII Th cells or Th cells with various gene deficiencies pre-stimulated in vitro by ovalbumin (OVA-pulsed dendritic cell (DCova. CTL survival was kinetically assessed in these mice using FITC-anti-CD8 and PE-H-2K(b/OVA257-264 tetramer staining by flow cytometry. We show that by acting via endogenous CD40L and IL-2, and acquired peptide-MHC-I (pMHC-I complex signaling, CD4(+ Th cells enhance survival of transferred effector CTLs and their differentiation into the functional memory CTLs capable of protecting against highly-metastasizing tumor challenge. Moreover, RT-PCR, flow cytometry and Western blot analysis demonstrate that increased survival of CD4(+ Th cell-helped CTLs is matched with enhanced Akt1/NF-κB activation, down-regulation of TRAIL, and altered expression profiles with up-regulation of prosurvival (Bcl-2 and down-regulation of proapoptotic (Bcl-10, Casp-3, Casp-4, Casp-7 molecules. Taken together, our results reveal a previously unexplored mechanistic role for CD4(+ Th cells in programming CTL survival and memory recall responses. This knowledge could also aid in the development of efficient adoptive CTL cancer therapy.

  8. Ethanol suppresses T cell proliferation without inhibiting interleukin 2 (IL2) production and IL2 receptor (IL2R) expression

    Energy Technology Data Exchange (ETDEWEB)

    Chang, M.P.; Norman, D.C. (VA Medical Center, West Los Angeles, CA (United States) Univ. of California, Los Angeles (United States))

    1991-03-11

    The effect of extended ethanol consumption of young C57BL/6J mice on T cell proliferation was studied. Splenic cells of young mice, fed with one of three different liquid diets for 6-7 weeks were cultured with Con A to assess T cell proliferation and production of IL2. Then, the proliferative response of splenic cells to PMA/ionomycin was assessed. Finally, Con A-activated T blast cells were assessed for their ability to express IL2R and to respond to IL2. The results showed that both Con A-induced mitogenesis and IL2-dependent proliferation of T cells from ethanol diet-fed mice were diminished as compared to that of maltose-substitute diet or standard liquid diet. However, the ability of T cells from ethanol diet-fed mice to produce IL2 and to express IL2R was not affected. Furthermore, the magnitude of ethanol-mediated suppression of T cell proliferation induced by PMA/ionomycin was comparable to that induced by Con A. These results taken together suggest that ethanol suppresses T cell proliferation by interfering with events following the IL2-IL2R interaction.

  9. IL-2 induces T cell adherence to extracellular matrix: inhibition of adherence and migration by IL-2 peptides generated by leukocyte elastase.

    Science.gov (United States)

    Ariel, A; Yavin, E J; Hershkoviz, R; Avron, A; Franitza, S; Hardan, I; Cahalon, L; Fridkin, M; Lider, O

    1998-09-01

    Migration of inflammatory cells requires cell adhesion and their subsequent detachment from the extracellular matrix (ECM). Leukocyte activation and migration must be terminated to stop inflammation. Here, we report that IL-2 enhances human T cell adherence to laminin, collagen type IV, and fibronectin (FN). In contrast, neutrophil elastase, an enzyme activated during inflammation, degrades IL-2 to yield IL-2 fractions that inhibit IL-2-induced T cell adhesion to FN. The amino acid composition of two of these IL-2 fractions, which appear to block T cell adherence to FN, were analyzed, and three peptides were consequently synthesized. The three peptides IVL, RMLT, and EFLNRWIT, but not the corresponding inversely synthesized peptides, inhibited T cell adhesion to FN induced by a variety of activators: IL-2, IL-7, macrophage inflammatory protein (MIP)-1beta, and PMA, as well as anti-CD3 and anti-beta1 integrin-activating mAb. Moreover, these IL-2 peptides inhibited T cell chemotaxis via FN-coated membranes induced by IL-2 and MIP-1beta. Inhibition of T cell adherence and migration apparently involves abrogation of the rearrangement of the T cell actin cytoskeleton. Thus, the migrating immune cells, the cytokines, and the ECM can create a functional relationship in which both inflammation-inducing signals and inhibitory molecules of immune responses can coexist; the enzymatic products of IL-2 may serve as natural feedback inhibitors of inflammation. PMID:9725245

  10. Con A-induced secretion of IL-2-like activity by mouse Thy-1+ epidermal cells

    International Nuclear Information System (INIS)

    Recently the authors reported that Con A stimulates in vitro proliferation of Thy-1+ dendritic epidermal cells (EC). They have not investigated the capacity of Con A-stimulated Thy-1+ EC to secrete IL-2-like activity (IL-2) 7-17EC is a 4 month-old line of Thy-1+ EC established from AKR/J EC by repeated stimulation with Con A and IL-2; these cells were 99% Thy-1+, but <1% L3T4+ and only 4-10% Lyt-2+. 7-17EC were harvested, washed, and plated in 96-well U-plates. Proliferation was assayed after 3H-thymidine pulsing; culture media was tested for IL-2 using the IL-2-dependent HT-2 cell line. With continuous Con A, marked IL-2 secretion (day 2 peak) was followed in parallel by proliferation (day 4 peak). In the absence of Con A, neither IL-2 nor proliferation was seen; however, both were recovered by addition of Con A to cells cultured first for 48 hr in media alone. 7-17EC proliferation was maximal at 0.5-2 μg Con A while maximal IL-2 secretion (100-200 U/ml) was seen at 4-8 μg/ml. That this amount of IL-2 was in excess of that required for their own optimal proliferation was supported by the failure of additional recombinant IL-2 (10 U/ml) to enhance Con A-stimulated proliferation of 7-17EC over a 4 day period. These findings further document the wide range of immunologic capabilities of Thy-1+ cells derived from epidermis

  11. Role of IL-2 in cancer immunotherapy.

    Science.gov (United States)

    Jiang, Tao; Zhou, Caicun; Ren, Shengxiang

    2016-06-01

    Interleukin-2 (IL-2) is one of the key cytokines with pleiotropic effects on immune system. It has been approved for the treatment of metastatic renal cell carcinoma and metastatic melanoma. Recent progress has been made in our understanding of IL-2 in regulating lymphocytes that has led to exciting new directions for cancer immunotherapy. While improved IL-2 formulations might be used as monotherapies, their combination with other anticancer immunotherapies, such as adoptive cell transfer regimens, antigen-specific vaccination, and blockade of immune checkpoint inhibitory molecules, for example cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) and programmed death 1 (PD-1) mono-antibodies, would held the promise of treating metastatic cancer. Despite the comprehensive studies of IL-2 on immune system have established the application of IL-2 for cancer immunotherapy, a number of poignant obstacles remain for future research. In the present review, we will focus on the key biological features of IL-2, current applications, limitations, and future directions of IL-2 in cancer immunotherapy. PMID:27471638

  12. Role of IL-2 in cancer immunotherapy.

    Science.gov (United States)

    Jiang, Tao; Zhou, Caicun; Ren, Shengxiang

    2016-06-01

    Interleukin-2 (IL-2) is one of the key cytokines with pleiotropic effects on immune system. It has been approved for the treatment of metastatic renal cell carcinoma and metastatic melanoma. Recent progress has been made in our understanding of IL-2 in regulating lymphocytes that has led to exciting new directions for cancer immunotherapy. While improved IL-2 formulations might be used as monotherapies, their combination with other anticancer immunotherapies, such as adoptive cell transfer regimens, antigen-specific vaccination, and blockade of immune checkpoint inhibitory molecules, for example cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) and programmed death 1 (PD-1) mono-antibodies, would held the promise of treating metastatic cancer. Despite the comprehensive studies of IL-2 on immune system have established the application of IL-2 for cancer immunotherapy, a number of poignant obstacles remain for future research. In the present review, we will focus on the key biological features of IL-2, current applications, limitations, and future directions of IL-2 in cancer immunotherapy.

  13. The IL-2/IL-2-Receptor Complex in the Maturation of Rat T-Cell Progenitors

    OpenAIRE

    Alberto Varas; Teresa Romo; Eva Jiménez; Luis Alonso; Angeles Vicente; Agustín G. Zapata

    1998-01-01

    On the basis of both the interleukin-2-receptor (IL-2R) α-chain expression on 16-day-old fetal rat thymocytes and the occurrence of interleukin-2 (IL-2) mRNA-containing cells early during rat thymus ontogeny, we have investigated the possible role of IL-2/IL-2R complex in rat T-cell maturation. For this purpose, we analyzed the effects of the addition of either recombinant rat IL- 2 or anti-CD25 (OX-39)-blocking monoclonal antibodies to fetal thymus organ cultures (FTOC), established from 16-...

  14. Selective targeting of IL-2 to NKG2D bearing cells for improved immunotherapy

    Science.gov (United States)

    Ghasemi, Reza; Lazear, Eric; Wang, Xiaoli; Arefanian, Saeed; Zheleznyak, Alexander; Carreno, Beatriz M.; Higashikubo, Ryuji; Gelman, Andrew E.; Kreisel, Daniel; Fremont, Daved H.; Krupnick, Alexander Sasha

    2016-01-01

    Despite over 20 years of clinical use, IL-2 has not fulfilled expectations as a safe and effective form of tumour immunotherapy. Expression of the high affinity IL-2Rα chain on regulatory T cells mitigates the anti-tumour immune response and its expression on vascular endothelium is responsible for life threatening complications such as diffuse capillary leak and pulmonary oedema. Here we describe the development of a recombinant fusion protein comprised of a cowpox virus encoded NKG2D binding protein (OMCP) and a mutated form of IL-2 with poor affinity for IL-2Rα. This fusion protein (OMCP-mutIL-2) potently and selectively activates IL-2 signalling only on NKG2D-bearing cells, such as natural killer (NK) cells, without broadly activating IL-2Rα-bearing cells. OMCP-mutIL-2 provides superior tumour control in several mouse models of malignancy and is not limited by mouse strain-specific variability of NK function. In addition, OMCP-mutIL-2 lacks the toxicity and vascular complications associated with parental wild-type IL-2. PMID:27650575

  15. Modulation of the tyrosine kinase receptor Ret/glial cell-derived neurotrophic factor (GDNF) signaling: a new player in reproduction induced anterior pituitary plasticity?

    Science.gov (United States)

    Guillou, Anne; Romanò, Nicola; Bonnefont, Xavier; Le Tissier, Paul; Mollard, Patrice; Martin, Agnès O

    2011-02-01

    During gestation, parturition, and lactation, the endocrine axis of the dam must continually adapt to ensure the continual and healthy development of offspring. The anterior pituitary gland, which serves as the endocrine interface between the brain and periphery, undergoes adaptations that contribute to regulation of the reproductive axis. Growth factors and their receptors are potential candidates for intrapituitary and paracrine factors to participate in the functional and anatomical plasticity of the gland. We examined the involvement of the growth factor glial cell-derived neurotrophic factor (GDNF) and its receptor tyrosine kinase rearranged during transfection (Ret) in the physiological functional and anatomical plasticity of the anterior pituitary gland. We found that variations in both expression and subcellular localization of Ret during gestation and lactation are temporally correlated with changes in pituitary gland function. We showed that Ret/GDNF signaling could endorse two different functional roles depending on the physiological status. At the end of lactation and after weaning, Ret was colocalized with markers of apoptosis. We found that Ret could therefore act as a physiological dependence receptor capable of inducing apoptosis in the absence of GDNF. In addition, we identified the follicullostellate cell as a probable source for intrapituitary GDNF and proposed GDNF as a potential physiological modulator of endocrine cell function. During all stages studied, we showed that acute application of GDNF to pituitary slices was able to modulate both positively and negatively intracellular calcium activity. Altogether our results implicate Ret/GDNF as a potent pleiotropic factor able to influence pituitary physiology during a period of high plasticity. PMID:21239429

  16. Nuclear Phosphoproteomic Screen Uncovers ACLY as Mediator of IL-2-induced Proliferation of CD4+ T lymphocytes.

    Science.gov (United States)

    Osinalde, Nerea; Mitxelena, Jone; Sánchez-Quiles, Virginia; Akimov, Vyacheslav; Aloria, Kerman; Arizmendi, Jesus M; Zubiaga, Ana M; Blagoev, Blagoy; Kratchmarova, Irina

    2016-06-01

    Anti-cancer immunotherapies commonly rely on the use of interleukin-2 (IL-2) to promote the expansion of T lymphocytes. IL-2- dependent proliferation is the culmination of a complex network of phosphorylation-driven signaling events that impact on gene transcription through mechanisms that are not clearly understood. To study the role of IL-2 in the regulation of nuclear protein function we have performed an unbiased mass spectrometry-based study of the nuclear phosphoproteome of resting and IL-2-treated CD4(+) T lymphocytes. We detected 8521distinct phosphosites including many that are not yet reported in curated phosphorylation databases. Although most phosphorylation sites remained unaffected upon IL-2 treatment, 391 sites corresponding to 288 gene products showed robust IL-2-dependent regulation. Importantly, we show that ATP-citrate lyase (ACLY) is a key phosphoprotein effector of IL-2-mediated T-cell responses. ACLY becomes phosphorylated on serine 455 in T lymphocytes upon IL-2-driven activation of AKT, and depletion or inactivation of ACLY compromises IL-2-promoted T-cell growth. Mechanistically, we demonstrate that ACLY is required for enhancing histone acetylation levels and inducing the expression of cell cycle regulating genes in response to IL-2. Thus, the metabolic enzyme ACLY emerges as a bridge between cytokine signaling and proliferation of T lymphocytes, and may be an attractive candidate target for the development of more efficient anti-cancer immunotherapies. PMID:27067055

  17. Nuclear Phosphoproteomic Screen Uncovers ACLY as Mediator of IL-2-induced Proliferation of CD4+ T lymphocytes.

    Science.gov (United States)

    Osinalde, Nerea; Mitxelena, Jone; Sánchez-Quiles, Virginia; Akimov, Vyacheslav; Aloria, Kerman; Arizmendi, Jesus M; Zubiaga, Ana M; Blagoev, Blagoy; Kratchmarova, Irina

    2016-06-01

    Anti-cancer immunotherapies commonly rely on the use of interleukin-2 (IL-2) to promote the expansion of T lymphocytes. IL-2- dependent proliferation is the culmination of a complex network of phosphorylation-driven signaling events that impact on gene transcription through mechanisms that are not clearly understood. To study the role of IL-2 in the regulation of nuclear protein function we have performed an unbiased mass spectrometry-based study of the nuclear phosphoproteome of resting and IL-2-treated CD4(+) T lymphocytes. We detected 8521distinct phosphosites including many that are not yet reported in curated phosphorylation databases. Although most phosphorylation sites remained unaffected upon IL-2 treatment, 391 sites corresponding to 288 gene products showed robust IL-2-dependent regulation. Importantly, we show that ATP-citrate lyase (ACLY) is a key phosphoprotein effector of IL-2-mediated T-cell responses. ACLY becomes phosphorylated on serine 455 in T lymphocytes upon IL-2-driven activation of AKT, and depletion or inactivation of ACLY compromises IL-2-promoted T-cell growth. Mechanistically, we demonstrate that ACLY is required for enhancing histone acetylation levels and inducing the expression of cell cycle regulating genes in response to IL-2. Thus, the metabolic enzyme ACLY emerges as a bridge between cytokine signaling and proliferation of T lymphocytes, and may be an attractive candidate target for the development of more efficient anti-cancer immunotherapies.

  18. Activation of the IL-2 Receptor in Podocytes: A Potential Mechanism for Podocyte Injury in Idiopathic Nephrotic Syndrome?

    Science.gov (United States)

    Zea, Arnold H.; Stewart, Tyrus; Ascani, Jeannine; Tate, David J.; Finkel-Jimenez, Beatriz; Wilk, Anna; Reiss, Krzysztof; Smoyer, William E.; Aviles, Diego H.

    2016-01-01

    The renal podocyte plays an important role in maintaining the structural integrity of the glomerular basement membrane. We have previously reported that patients with idiopathic nephrotic syndrome (INS) have increased IL-2 production. We hypothesized that podocytes express an IL-2 receptor (IL-2R) and signaling through this receptor can result in podocyte injury. To confirm the presence of the IL-2R, we tested a conditionally immortalized murine podocyte cell line by flow cytometry, qPCR, and Western blot. To test for the presence of the IL-2R in vivo, immunohistochemical staining was performed on human renal biopsies in children with FSGS and control. Podocytes were stimulated with IL-2 in vitro, to study signaling events via the JAK/STAT pathway. The results showed that stimulation with IL-2 resulted in increased mRNA and protein expression of STAT 5a, phosphorylated STAT 5, JAK 3, and phosphorylated JAK 3. We then investigated for signs of cellular injury and the data showed that pro-apoptotic markers Bax and cFLIP were significantly increased following IL-2 exposure, whereas LC3 II was decreased. Furthermore, mitochondrial depolarization and apoptosis were both significantly increased following activation of the IL-2R. We used a paracellular permeability assay to monitor the structural integrity of a podocyte monolayer following IL-2 exposure. The results showed that podocytes exposed to IL-2 have increased albumin leakage across the monolayer. We conclude that murine podocytes express the IL-2R, and that activation through the IL-2R results in podocyte injury. PMID:27389192

  19. Activation of the IL-2 Receptor in Podocytes: A Potential Mechanism for Podocyte Injury in Idiopathic Nephrotic Syndrome?

    Directory of Open Access Journals (Sweden)

    Arnold H Zea

    Full Text Available The renal podocyte plays an important role in maintaining the structural integrity of the glomerular basement membrane. We have previously reported that patients with idiopathic nephrotic syndrome (INS have increased IL-2 production. We hypothesized that podocytes express an IL-2 receptor (IL-2R and signaling through this receptor can result in podocyte injury. To confirm the presence of the IL-2R, we tested a conditionally immortalized murine podocyte cell line by flow cytometry, qPCR, and Western blot. To test for the presence of the IL-2R in vivo, immunohistochemical staining was performed on human renal biopsies in children with FSGS and control. Podocytes were stimulated with IL-2 in vitro, to study signaling events via the JAK/STAT pathway. The results showed that stimulation with IL-2 resulted in increased mRNA and protein expression of STAT 5a, phosphorylated STAT 5, JAK 3, and phosphorylated JAK 3. We then investigated for signs of cellular injury and the data showed that pro-apoptotic markers Bax and cFLIP were significantly increased following IL-2 exposure, whereas LC3 II was decreased. Furthermore, mitochondrial depolarization and apoptosis were both significantly increased following activation of the IL-2R. We used a paracellular permeability assay to monitor the structural integrity of a podocyte monolayer following IL-2 exposure. The results showed that podocytes exposed to IL-2 have increased albumin leakage across the monolayer. We conclude that murine podocytes express the IL-2R, and that activation through the IL-2R results in podocyte injury.

  20. Abrogation of CD40–CD154 Signaling Impedes the Homeostasis of Thymic Resident Regulatory T Cells by Altering the Levels of IL-2, but Does Not Affect Regulatory T Cell Development

    OpenAIRE

    Cuss, Steven M.; Green, E. Allison

    2012-01-01

    Identification of costimulatory signals required for murine regulatory T (Treg) cell development relies on measuring the frequency of total thymic Treg cells. However, the thymus contains both resident and newly developed Treg cells; whether such signals target both populations is unknown. In this study, we show that CD40–CD154 blockade specifically targeted thymic resident Treg cells, but not, as was previously believed, newly developed Treg cells. Unlike CD28–CD80/CD86 signals, CD40–CD154 s...

  1. EBI2 augments Tfh cell fate by promoting interaction with IL-2-quenching dendritic cells.

    Science.gov (United States)

    Li, Jianhua; Lu, Erick; Yi, Tangsheng; Cyster, Jason G

    2016-05-01

    T follicular helper (Tfh) cells are a subset of T cells carrying the CD4 antigen; they are important in supporting plasma cell and germinal centre responses. The initial induction of Tfh cell properties occurs within the first few days after activation by antigen recognition on dendritic cells, although how dendritic cells promote this cell-fate decision is not fully understood. Moreover, although Tfh cells are uniquely defined by expression of the follicle-homing receptor CXCR5 (refs 1, 2), the guidance receptor promoting the earlier localization of activated T cells at the interface of the B-cell follicle and T zone has been unclear. Here we show that the G-protein-coupled receptor EBI2 (GPR183) and its ligand 7α,25-dihydroxycholesterol mediate positioning of activated CD4 T cells at the interface of the follicle and T zone. In this location they interact with activated dendritic cells and are exposed to Tfh-cell-promoting inducible co-stimulator (ICOS) ligand. Interleukin-2 (IL-2) is a cytokine that has multiple influences on T-cell fate, including negative regulation of Tfh cell differentiation. We demonstrate that activated dendritic cells in the outer T zone further augment Tfh cell differentiation by producing membrane and soluble forms of CD25, the IL-2 receptor α-chain, and quenching T-cell-derived IL-2. Mice lacking EBI2 in T cells or CD25 in dendritic cells have reduced Tfh cells and mount defective T-cell-dependent plasma cell and germinal centre responses. These findings demonstrate that distinct niches within the lymphoid organ T zone support distinct cell fate decisions, and they establish a function for dendritic-cell-derived CD25 in controlling IL-2 availability and T-cell differentiation.

  2. Changes in Gab2 phosphorylation and interaction partners in response to interleukin (IL)-2 stimulation in T-lymphocytes

    DEFF Research Database (Denmark)

    Osinalde, Nerea; Sánchez-Quiles, Virginia; Blagoev, Blagoy;

    2016-01-01

    Interleukin-2 (IL-2) stimulation results in T-cell growth as a consequence of activation of highly sophisticated and fine-tuned signaling pathways. Despite lacking intrinsic enzymatic activity, scaffold proteins such as Gab2, play a pivotal role in IL-2-triggered signal transduction integrating......-lymphocytes and besides well-known Gab2 interactors we discover three novel cytokine-inducible Gab2-binding proteins. Thus, our data provide novel insights and a wealth of candidates for future studies that will shed light into the role of Gab2 in IL-2-initiated signal transduction....

  3. Hybrid Cells Derived from Human Breast Cancer Cells and Human Breast Epithelial Cells Exhibit Differential TLR4 and TLR9 Signaling

    Directory of Open Access Journals (Sweden)

    Songül Tosun

    2016-05-01

    Full Text Available TLRs are important receptors of cells of the innate immune system since they recognize various structurally conserved molecular patterns of different pathogens as well as endogenous ligands. In cancer, the role of TLRs is still controversial due to findings that both regression and progression of tumors could depend on TLR signaling. In the present study, M13SV1-EGFP-Neo human breast epithelial cells, MDA-MB-435-Hyg human breast cancer cells and two hybrids M13MDA435-1 and -3 were investigated for TLR4 and TLR9 expression and signaling. RT-PCR data revealed that LPS and CpG-ODN induced the expression of pro-inflammatory cytokines, like IFN-β, TNF-α, IL-1β and IL-6 in hybrid cells, but not parental cells. Interestingly, validation of RT-PCR data by Western blot showed detectable protein levels solely after LPS stimulation, suggesting that regulatory mechanisms are also controlled by TLR signaling. Analysis of pAKT and pERK1/2 levels upon LPS and CpG-ODN stimulation revealed a differential phosphorylation pattern in all cells. Finally, the migratory behavior of the cells was investigated showing that both LPS and CpG-ODN potently blocked the locomotory activity of the hybrid cells in a dose-dependent manner. In summary, hybrid cells exhibit differential TLR4 and TLR9 signaling.

  4. Atoh7 promotes the differentiation of retinal stem cells derived from Müller cells into retinal ganglion cells by inhibiting Notch signaling

    OpenAIRE

    Song, Wei-tao; Zhang, Xue-yong; Xia, Xiao-Bo

    2013-01-01

    Introduction Retinal Müller cells exhibit the characteristics of retinal progenitor cells, and differentiate into ganglion cells under certain conditions. However, the number of ganglion cells differentiated from retinal Müller cells falls far short of therapeutic needs. This study aimed to develop a novel protocol to promote the differentiation of retinal Müller cells into ganglion cells and explore the underlying signaling mechanisms. Methods Müller cells were isolated and purified from rat...

  5. Nuclear Phosphoproteomic Screen Uncovers ACLY as Mediator of IL-2-induced Proliferation of CD4+ T lymphocytes

    DEFF Research Database (Denmark)

    Osinalde, Nerea; Mitxelena, Jone; Sánchez-Quiles, Virginia;

    2016-01-01

    Anti-cancer immunotherapies commonly rely on the use of interleukin-2 (IL-2) to promote the expansion of T lymphocytes. IL-2- dependent proliferation is the culmination of a complex network of phosphorylation-driven signaling events that impact on gene transcription through mechanisms...... and inducing the expression of cell cycle regulating genes in response to IL-2. Thus, the metabolic enzyme ACLY emerges as a bridge between cytokine signaling and proliferation of T lymphocytes, and may be an attractive candidate target for the development of more efficient anti-cancer immunotherapies....

  6. Specification of embryonic stem cell-derived tissues into eye fields by Wnt signaling using rostral diencephalic tissue-inducing culture.

    Science.gov (United States)

    Sakakura, Eriko; Eiraku, Mototsugu; Takata, Nozomu

    2016-08-01

    The eyes are subdivided from the rostral diencephalon in early development. How the neuroectoderm regulates this subdivision, however, is largely unknown. Taking advantage of embryonic stem cell (ESC) culture using a Rax reporter line to monitor rostral diencephalon formation, we found that ESC-derived tissues at day 7 grown in Glasgow Minimum Expression Media (GMEM) containing knockout serum replacement (KSR) exhibited higher levels of expression of axin2, a Wnt target gene, than those grown in chemically defined medium (CDM). Surprisingly, Wnt agonist facilitated eye field-like tissue specification in CDM. In contrast, the addition of Wnt antagonist diminished eye field tissue formation in GMEM+KSR. Furthermore, the morphological formation of the eye tissue anlage, including the optic vesicle, was accompanied by Wnt signaling activation. Additionally, using CDM culture, we developed an efficient method for generating Rax+/Chx10+ retinal progenitors, which could become fully stratified retina. Here we provide a new avenue for exploring the mechanisms of eye field specification in vitro.

  7. Autophagy is required for IL-2-mediated fibroblast growth

    Energy Technology Data Exchange (ETDEWEB)

    Kang, Rui [Department of Surgery, University of Pittsburgh, Pittsburgh, Pennsylvania 15219 (United States); Tang, Daolin, E-mail: tangd2@upmc.edu [Department of Surgery, University of Pittsburgh, Pittsburgh, Pennsylvania 15219 (United States); Lotze, Michael T., E-mail: lotzemt@upcm.edu [Department of Surgery, University of Pittsburgh, Pittsburgh, Pennsylvania 15219 (United States); Zeh III, Herbert J., E-mail: zehh@upmc.edu [Department of Surgery, University of Pittsburgh, Pittsburgh, Pennsylvania 15219 (United States)

    2013-02-15

    Autophagy is an evolutionarily conserved pathway responsible for delivery of cytoplasmic material into the lysosomal degradation pathway to enable vesicular exocytosis. Interleukin (IL)-2 is produced by T-cells and its activity is important for immunoregulation. Fibroblasts are an immune competent cell type, playing a critical role in wound healing, chronic inflammation, and tumor development. Although autophagy plays an important role in each of these processes, whether it regulates IL-2 activity in fibroblasts is unknown. Here, we show that autophagy is required for IL-2-induced cell growth in fibroblasts. IL-2 significantly induced autophagy in mouse embryonic fibroblasts (MEFs) and primary lung fibroblasts. Autophagy inhibitors (e.g., 3-methylamphetamine and bafilomycin A1) or knockdown of ATG5 and beclin 1 blocked clinical grade IL-2-induced autophagy. Moreover, IL-2 induced HMGB1 cytoplasmic translocation in MEFs and promoted interaction between HMGB1 and beclin1, which is required for autophagy induction. Pharmacological and genetic inhibition of autophagy inhibited IL-2-induced cell proliferation and enhanced IL-2-induced apoptosis. These findings suggest that autophagy is an important pro-survival regulator for IL-2-induced cell growth in fibroblasts.

  8. Transcriptional regulation of IL-2 in health and autoimmunity

    Science.gov (United States)

    Crispín, José C.; Tsokos, George C.

    2009-01-01

    The regulation of IL-2 production is central to our understanding of the immune system. Key during T cell activation, it also plays an essential role in the regulation of the immune response. This review discusses the function of recently described factors that modulate transcription and chromatin remodeling at the IL2 promoter. Also, it addresses the role of FoxP3 as a transcriptional regulator in conventional T cells and regulatory T cells, and the mechanisms whereby CD28 stabilizes IL2 transcription and translation. Finally, the alterations that prevent T cells from SLE patients from producing normal amounts of IL-2 upon stimulation are described. PMID:18723131

  9. Locoregional IL-2 low dose applications for gastrointestinal tumors

    Institute of Scientific and Technical Information of China (English)

    Zachary Krastev; Willem Den Otter; V Koltchakov; R Tomova; S Deredjian; A Alexiev; D Popov; B Tomov; Jan-Willem Koten; John Jacobs

    2005-01-01

    AIM: To explore the feasibility of local interleukin 2 (IL-2)in patients with different forms of abdominal cancer. This required experimentation with the time interval between IL-2 applications and the methods of application.METHODS: Sixteen patients with stages Ⅲ and Ⅳ of gastrointestinal malignancies (primary or metastatic) who were admitted to our Department of Gastroenterology were treated with locoregionally applied IL-2 in low doses.RESULTS: No major problems applying locoregional IL-2 were encountered. In 6 out of 16 patients, a modest but clinically worthwhile improvement was obtained. Adverse effects were minimal, The therapeutic scheme was well tolerated, even in patients in a poor condition.CONCLUSION: This study demonstrates the feasibility of low dose locoregional IL-2 application in advanced abdominal cancer. Local IL-2 therapy gives only negligible adverse effects. The results suggest that it is important to apply intratumorally. Local IL-2 may be given adjunct to standard therapeutic regimes and does not imply complex surgical interventions. These initial results are encouraging.

  10. Changes in Gab2 phosphorylation and interaction partners in response to interleukin (IL)-2 stimulation in T-lymphocytes.

    Science.gov (United States)

    Osinalde, Nerea; Sánchez-Quiles, Virginia; Blagoev, Blagoy; Kratchmarova, Irina

    2016-01-01

    Interleukin-2 (IL-2) stimulation results in T-cell growth as a consequence of activation of highly sophisticated and fine-tuned signaling pathways. Despite lacking intrinsic enzymatic activity, scaffold proteins such as Gab2, play a pivotal role in IL-2-triggered signal transduction integrating, diversifying and amplifying the signal by serving as a platform for the assembly of effectors proteins. Traditionally, Gab2-mediated protein recruitment was believed to solely depend on cytokine-induced phosphotyrosine moieties. At present, phosphorylation on serine/threonine residues is also emerging as a key mediator of Gab2-dependent signal regulation. Despite its relevance, IL-2-triggered regulation on Gab2 phosphorylation is yet poorly understood. Combining antibody- and TiO2-based enrichment of the scaffold protein with SILAC quantitative mass spectrometry we disclose the prominent regulation IL-2 exerts on Gab2 serine/threonine phosphorylation by showing that at least 18 serines and 1 threonine, including previously non-reported ones, become phosphorylated in response to cytokine stimulation. Additionally, we decipher the interactome of the docking protein in resting and cytokine-treated T-lymphocytes and besides well-known Gab2 interactors we discover three novel cytokine-inducible Gab2-binding proteins. Thus, our data provide novel insights and a wealth of candidates for future studies that will shed light into the role of Gab2 in IL-2-initiated signal transduction. PMID:27025927

  11. Interleukin 2 (IL 2) inhibitor in rheumatoid synovial fluid: Correlation with prognosis and soluble IL 2 receptor levels

    International Nuclear Information System (INIS)

    A soluble activity inhibiting over 50% of the CTLL-2 cell line response to recombinant human interleukin 2 (IL 2) was found in 17 of 29 (59%) rheumatoid synovial fluids. To study the prognosis value of this activity, 16 rheumatoid synovial fluids were collected before a radiation synovectomy of the knee with 7 mCi of 90Y. Patients with a good clinical result after the synovectomy had a lower IL 2 inhibitory activity than those with a bad or incomplete result (P less than 0.01). Levels of inhibitory activity and of soluble IL 2 receptors were correlated with each other and with the response of the synovitis to the radiation synovectomy. These results extend the clinical usefulness of soluble IL 2 receptor measurements and indicate a correlation between the immune activation of the rheumatoid synovitis and its clinical activity

  12. Phase I clinical trial combining imatinib mesylate and IL-2

    Science.gov (United States)

    Chaput, Nathalie; Flament, Caroline; Locher, Clara; Desbois, Mélanie; Rey, Annie; Rusakiewicz, Sylvie; Poirier-Colame, Vichnou; Pautier, Patricia; Le Cesne, Axel; Soria, Jean-Charles; Paci, Angelo; Rosenzwajg, Michelle; Klatzmann, David; Eggermont, Alexander; Robert, Caroline; Zitvogel, Laurence

    2013-01-01

    We performed a Phase I clinical trial from October 2007 to October 2009, enrolling patients affected by refractory solid tumors, to determine the maximum tolerated dose (MTD) of interleukin (IL)-2 combined with low dose cyclophosphamide (CTX) and imatinib mesylate (IM). In a companion paper published in this issue of OncoImmunology, we show that the MTD of IL-2 is 6 MIU/day for 5 consecutive days, and that IL-2 increases the impregnation of both IM and of its main metabolite, CGP74588. Among the secondary objectives, we wanted to determine immunological markers that might be associated with progression-free survival (PFS) and/or overall survival (OS). The combination therapy markedly reduced the absolute counts of B, CD4+ T and CD8+ T cells in a manner that was proportional to IL-2 dose. There was a slight (less than 2-fold) increase in the proportion of regulatory T cells (Tregs) among CD4+ T cells in response to IM plus IL-2. The natural killer (NK)-cell compartment was activated, exhibiting a significant upregulation of HLA-DR, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and CD56. The abundance of HLA-DR+ NK cells after one course of combination therapy positively correlated with both PFS and OS. The IL-2-induced rise of the CD4+:CD8+ T-cell ratio calculated after the first cycle of treatment was also positively associated with OS. Overall, the combination of IM and IL-2 promoted the rapid expansion of HLA-DR+ NK cells and increased the CD4+:CD8+ T-cell ratio, both being associated with clinical benefits. This combinatorial regimen warrants further investigation in Phase II clinical trials, possibly in patients affected by gastrointestinal stromal tumors, a setting in which T and NK cells may play an important therapeutic role. PMID:23525357

  13. Study on the construction of recombinant plasmid coexpressing newcastle disease virus F protein and chicken IL-2

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    This study investigated the protection against the ND in chickens by a recombinant DNA vaccine. A plasmid vector encoding NDV F protein, which is reqired for virus cell fusion and is important for vaccine induced immunity, was used as a model to study how DNA vaccines may be modulated by the simulaneous expression of chicken IL-2. The NDV D26 strain F gene with CMV promotor and BGH polyA signal sequence was amplified by PCR from eukaryotic plasmid pcDNA-F, which contains the full-length NDV F gene, and clond into reconstructed eukaryotic plasmid pcDNA-IL2, which contains chicken IL-2 gene. Restriction endonuclease cleavage and PCR amplification showed that a bicistronic plasmid encoding NDV F gene and chicken IL-2 separately was successfully constructed. Two-week-old SPF chickens were intramuscularly innoculated the recombinant plasmid. Antibody and lymphocyte proliferative assay showed that the humoral and cellular immunity of chickens vaccinated the recombinant plasmid greatly increased compared with those innoculated only plasmid expressing NDV F protein. Challenged with the lethal dose of NDV F48E9 strain, 72% chickens vaccinated recombinant plasmid were survived, and 30% chickens vaccinated plasmid expressing F protein were survived. These results proved the adjuvant effect of chicken IL-2, and further showed that the efficacy of a DNA vaccine can be greatly improved by simultaneous expression of IL-2.

  14. Antigen-specific regulatory T cells and low dose of IL-2 in treatment of type 1 diabetes

    Directory of Open Access Journals (Sweden)

    Minh N. Pham

    2016-01-01

    Full Text Available Regulatory T cells (Tregs play an important role in preventing effector T-cell (Teff targeting of self-antigens that can lead to tissue destruction in autoimmune settings, including type 1 diabetes (T1D. Autoimmunity is caused in part by an imbalance between Teff and Tregs. Early attempts to treat with immunosuppressive agents have led to serious side effects, thus requiring a more targeted approach. Low-dose IL-2 (LD IL-2 can provide immuno-regulation with few side effects by preferentially acting on Tregs to drive tolerance. The concept of LD IL-2 as a therapeutic approach is supported by data in mouse models where autoimmunity is cured and further strengthened by success in human clinical studies in Hepatitis C Virus (HCV induced vasculitis, chronic graft vs host disease (GVHD and Alopecia areata (AA. Treatment will require identification of a safe therapeutic window, which is a difficult task given that patients are reported to have deficient or defective IL-2 production or signalling and have experienced mild activation of NK cells and eosinophils with LD IL-2 therapy. In T1D, a LD IL-2 clinical trial concluded that Tregs can be safely expanded in humans; however, the study was not designed to address efficacy. Antigen-specific therapies have also aimed at regulation of the autoimmune response, but have been filled with disappointment despite an extensive list of diverse islet antigens tested in humans. This approach could be enhanced through the addition of LD IL-2 to the antigenic treatment regimen to improve the frequency and function of antigen-specific Tregs, without global immunosuppression. Here we will discuss the use of LD IL-2 and islet antigen to enhance antigen-specific Tregs in T1D and focus on what is known about their immunological impact, their safety and potential efficacy, and need for better methods to identify therapeutic effectiveness.

  15. Suppression of Murine Allergic Airway Disease by IL-2:Anti-IL-2 Monoclonal Antibody-Induced Regulatory T Cells1

    OpenAIRE

    Wilson, Mark S; Pesce, John T; Thirumalai R Ramalingam; Thompson, Robert W.; Cheever, Allen; Wynn, Thomas A

    2008-01-01

    Regulatory T cells (Treg) play a decisive role in many diseases including asthma and allergen-induced lung inflammation. However, little progress has been made developing new therapeutic strategies for pulmonary disorders. In the current study we demonstrate that cytokine:antibody complexes of IL-2 and anti-IL-2 mAb reduce the severity of allergen-induced inflammation in the lung by expanding Tregs in vivo. Unlike rIL-2 or anti-IL-2 mAb treatment alone, IL-2:anti-IL-2 complexes dampened airwa...

  16. Local administration of cells containing an inserted IL-2 gene and producing IL-2 inhibits growth of human tumours in nu/nu mice.

    Science.gov (United States)

    Bubenik, J; Voitenok, N N; Kieler, J; Prassolov, V S; Chumakov, P M; Bubenikova, D; Simova, J; Jandlova, T

    1988-12-01

    We have prepared a retroviral expression construct, pPS-IL-2, in which human IL-2 cDNA has been inserted into the polylinker region, and have used the retroviral vector to introduce the functional IL-2 gene into a fibroblast cell line, RAT-1. Peritumoral administration of IL-2-producing RAT-1 cells into congenitally athymic (nu/nu) mice carrying subcutaneous transplants of human carcinoma cells inhibited the growth of the human tumour xenografts.

  17. Regulation of IL-2 gene expression by Siva and FOXP3 in human T cells

    Directory of Open Access Journals (Sweden)

    Hench Virginia K

    2011-09-01

    Full Text Available Abstract Background Severe autoinflammatory diseases are associated with mutations in the Foxp3 locus in both mice and humans. Foxp3 is required for the development, function, and maintenance of regulatory T cells (Tregs, a subset of CD4 cells that suppress T cell activation and inflammatory processes. Siva is a pro-apoptotic gene that is expressed across a range of tissues, including CD4 T cells. Siva interacts with three tumor necrosis factor receptor (TNFR family members that are constitutively expressed on Treg cells: CD27, GITR, and OX40. Results Here we report a biophysical interaction between FOXP3 and Siva. We mapped the interaction domains to Siva's C-terminus and to a central region of FOXP3. We showed that Siva repressed IL-2 induction by suppressing IL-2 promoter activity during T cell activation. Siva-1's repressive effect on IL-2 gene expression appears to be mediated by inhibition of NFkappaB, whereas FOXP3 repressed both NFkappaB and NFAT activity. Conclusions In summary, our data suggest that both FOXP3 and Siva function as negative regulators of IL-2 gene expression in Treg cells, via suppression of NFAT by FOXP3 and of NFkappaB by both FOXP3 and Siva. Our work contributes evidence for Siva's role as a T cell signalling mediator in addition to its known pro-apoptotic function. Though further investigations are needed, evidence for the biophysical interaction between FOXP3 and Siva invites the possibility that Siva may be important for proper Treg cell function.

  18. Interleukin 2 (IL 2) up-regulates its own receptor on a subset of human unprimed peripheral blood lymphocytes and triggers their proliferation

    International Nuclear Information System (INIS)

    Several reports indicate that human peripheral blood lymphoctyes (PBL) seeded in culture with purified or recombinant interleukin 2 (IL 2) immediately after separation from the blood display a substantial level of proliferation at day 5 or 6, even in the absence of any activating signal. The spontaneously IL 2 proliferating cells are large lymphocytes, and they co-purify on a Percoll gradient in the large granular lymphocytes (third (LGL) fraction) together with the natural killer (NK) activity. When LGL were separated into NKH1 (an NK-specific surface marker)-positive and NKH1-negative cells by fluorescence-activated cell sorting (FACS), proliferating cells were mainly found in the NKH1-negative fraction. On the contrary, when cells from Percoll fraction 3 were separated into OKT3-negative and positive cells, the majority of the proliferating cells was found in the OKT3-positive cells. These results indicate that spontaneously IL 2 proliferating (SIP) cells most probably belong to the T cell lineage, but are distinct from NK cells. Additional analysis of Il 2 receptor induced in culture with IL 2 was performed by [125I]anti-TAC binding and by [3H]Il 2 binding. Scatchard analysis of [3H]IL 2 binding, in the range of concentrations leading to the detection of high-affinity binding sites, showed an affinity constant similar to that of conventional phytohemagglutinin blasts. The results indicate that SIP cells are preactivated cells circulating in the blood. They are large cells and represent a very small proportion of circulating lymphocytes (0.3%). They express a subliminar amount of IL 2 receptor. Cultivated in the presence of IL 2, IL 2 receptor expression is enhanced to a detectable level, and the SIP cells begin to proliferate. These SIP cells could be activated T cells in the course of a current immune response or memory T cells present in every normal individual

  19. SILAC-based quantification of changes in protein tyrosine phosphorylation induced by Interleukin-2 (IL-2 and IL-15 in T-lymphocytes

    Directory of Open Access Journals (Sweden)

    Nerea Osinalde

    2015-12-01

    Full Text Available This data article presents the first large-scale quantitative phosphoproteomics dataset generated to decipher the signaling networks initiated by IL-2 and IL-15 in T-lymphocytes. Data was collected by combining immunoprecipitation of tyrosine phosphorylated proteins and TiO2-based phosphopeptide enrichment with SILAC-based quantitative mass spectrometry. We report all the proteins and phosphotyrosine-containing peptides identified and quantified in IL-2- and IL-15-stimulated T-lymphocytes. The gene ontology analysis of IL-2 and IL-15 effector proteins detected in the present work is also included. The data supplied in this article is related to the research work entitled “Simultaneous dissection and comparison of IL-2 and IL-15 signaling pathways by global quantitative phosphoproteomics” [1]. All mass spectrometry data have been deposited in the ProteomeXchange with the identifier PXD001129.

  20. 脐血IL-2,mIL-2R,sIL-2R和T细胞免疫功能%Expression levels of IL-2, sIL-2R and mIL-2R of cord blood and the immune function of T cells

    Institute of Scientific and Technical Information of China (English)

    许礼发; 王健; 李朝品

    2004-01-01

    1实验资料ELISA法IL-2、可溶性白细胞介素-2受体(soluble interleukin-2 receptor,sIL-2R),IL-6,IL-10和可溶性细胞间黏附分子-1(soluble intercellular adhesion molecule-1,sICAM-1)检测试剂盒均购自法国Dia Clone公司和美国Biosource公司,PHA为广州医药公司产品,生物素-链霉亲和素(biotin-streptavidin,BSA)法膜白细胞介素-2受体(membrane

  1. Stem Cell-Derived Extracellular Vesicles and Immune-Modulation.

    Science.gov (United States)

    Burrello, Jacopo; Monticone, Silvia; Gai, Chiara; Gomez, Yonathan; Kholia, Sharad; Camussi, Giovanni

    2016-01-01

    Extra-cellular vesicles (EVs) are bilayer membrane structures enriched with proteins, nucleic acids, and other active molecules and have been implicated in many physiological and pathological processes over the past decade. Recently, evidence suggests EVs to play a more dichotomic role in the regulation of the immune system, whereby an immune response may be enhanced or supressed by EVs depending on their cell of origin and its functional state. EVs derived from antigen (Ag)-presenting cells for instance, have been involved in both innate and acquired (or adaptive) immune responses, as Ag carriers or presenters, or as vehicles for delivering active signaling molecules. On the other hand, tumor and stem cell derived EVs have been identified to exert an inhibitory effect on immune responses by carrying immuno-modulatory effectors, such as transcriptional factors, non-coding RNA (Species), and cytokines. In addition, stem cell-derived EVs have also been reported to impair dendritic cell maturation and to regulate the activation, differentiation, and proliferation of B cells. They have been shown to control natural killer cell activity and to suppress the innate immune response (IIR). Studies reporting the role of EVs on T lymphocyte modulation are controversial. Discrepancy in literature may be due to stem cell culture conditions, methods of EV purification, EV molecular content, and functional state of both parental and target cells. However, mesenchymal stem cell-derived EVs were shown to play a more suppressive role by shifting T cells from an activated to a T regulatory phenotype. In this review, we will discuss how stem cell-derived EVs may contribute toward the modulation of the immune response. Collectively, stem cell-derived EVs mainly exhibit an inhibitory effect on the immune system. PMID:27597941

  2. LPS nephropathy in mice is ameliorated by IL-2 independently of regulatory T cells activity.

    Directory of Open Access Journals (Sweden)

    Roberta Bertelli

    Full Text Available Immunosuppressive regulatory T cells (Tregs have been hypothesized to exert a protective role in animal models of spontaneous (Buffalo/Mna and/or drug induced (Adriamycin nephrotic syndrome. In this study, we thought to define whether Tregs can modify the outcome of LPS nephropathy utilizing IL-2 as inducer of tissue and circulating Tregs. LPS (12 mg/Kg was given as single shot in C57BL/6, p2rx7⁻/⁻ and Foxp3EGFP; free IL-2 (18.000 U or, in alternative, IL-2 coupled with JES6-1 mAb (IL-2/anti-IL-2 were injected before LPS. Peripheral and tissue Tregs/total CD4+ cell ratio, urinary parameters and renal histology were evaluated for 15 days. IL-2 administration to wild type mice had no effect on peripheral Tregs number, whereas a significant increase was induced by the IL-2/anti-IL-2 immunocomplex after 5 days. Spleen and lymph nodes Tregs were comparably increased. In p2rx7⁻/⁻ mice, IL-2/anti-IL-2 treatment resulted in increase of peripheral Tregs but did not modify the spleen and lymph nodes quota. LPS induced comparable and transient proteinuria in both wild type and p2rx7⁻/⁻ mice. Proteinuria was inhibited by co-infusion of human IL-2, with reduction at each phase of the disease (24 -48 and 72 hours whereas IL-2/anti-IL-2 produced weaker effects. In all mice (wild type and p2rx7⁻/⁻ and irrespective of treatment (IL-2, IL-2/anti-IL-2, LPS was associated with progressive signs of renal pathologic involvement resulting in glomerulosclerosis. In conclusion, IL-2 plays a transient protective effect on proteinuria induced by LPS independent of circulating or tissue Tregs but does not modify the outcome of renal degenerative renal lesions.

  3. New Molecular and Cellular Mechanisms of Tolerance: Tolerogenic Actions of IL-2.

    Science.gov (United States)

    Pérol, Louis; Piaggio, Eliane

    2016-01-01

    Interleukin-2 (IL-2) is an old molecule with brand new functions. Indeed, IL-2 has been first described as a T-cell growth factor but recent data pointed out that its main function in vivo is the maintenance of immune tolerance. Mechanistically, IL-2 is essential for the development and function of CD4(+) Foxp3(+) regulatory T cells (Treg cells) that are essential players in the control of immune responded to self, tumors, microbes and grafts. Treg cells are exquisitely sensitive to IL-2 due to their constitutive expression of the high affinity IL-2 receptor (IL-2R) and the new paradigm suggests that low-doses of IL-2 could selectively boost Treg cells in vivo. Consequently, a growing body of clinical research is aiming at using IL-2 at low doses as a tolerogenic drug to boost endogenous Treg cells in patients suffering from autoimmune or inflammatory conditions. In this manuscript, we briefly review IL-2/IL-2R biology and the role of IL-2 in the development, maintenance, and function of Treg cells; and also its effects on other immune cell populations such as CD4(+) T helper cells and CD8(+) memory T cells. Then, focusing on type 1 diabetes, we review the preclinical studies and clinical trials supporting the use of low-doses IL-2 as a tolerogenic immunotherapy. Finally, we discuss the limitations and future directions for IL-2 based immunotherapy. PMID:26530792

  4. Inability of a Fusion Protein of IL-2 and Diphtheria Toxin (Denileukin Diftitox, DAB389IL-2, ONTAK) to Eliminate Regulatory T Lymphocytes in Patients With Melanoma

    OpenAIRE

    Attia, Peter; Maker, Ajay V; Haworth, Leah R.; Rogers-Freezer, Linda; Rosenberg, Steven A.

    2005-01-01

    Elimination of regulatory T lymphocytes may provide a way to break self-tolerance and unleash the anti-tumor properties of circulating lymphocytes. The use of fusion proteins, which link cytotoxic molecules to receptor targets, provides one approach to this problem. This study examined the ability of a fusion protein of interleukin-2 (IL-2) and diphtheria toxin (Denileukin Diftitox, DAB389IL-2, ONTAK) to eliminate regulatory T lymphocytes based on their expression of high-affinity IL-2 recept...

  5. Placenta-specific Expression of the Interleukin-2 (IL-2) Receptor β Subunit from an Endogenous Retroviral Promoter*

    Science.gov (United States)

    Cohen, Carla J.; Rebollo, Rita; Babovic, Sonja; Dai, Elizabeth L.; Robinson, Wendy P.; Mager, Dixie L.

    2011-01-01

    The long terminal repeat (LTR) sequences of endogenous retroviruses and retroelements contain promoter elements and are known to form chimeric transcripts with nearby cellular genes. Here we show that an LTR of the THE1D retroelement family has been domesticated as an alternative promoter of human IL2RB, the gene encoding the β subunit of the IL-2 receptor. The LTR promoter confers expression specifically in the placental trophoblast as opposed to its native transcription in the hematopoietic system. Rather than sequence-specific determinants, DNA methylation was found to regulate transcription initiation and splicing efficiency in a tissue-specific manner. Furthermore, we detected the cytoplasmic signaling domain of the IL-2Rβ protein in the placenta, suggesting that IL-2Rβ undergoes preferential proteolytic cleavage in this tissue. These findings implicate novel functions for this cytokine receptor subunit in the villous trophoblast and reveal an intriguing example of ancient LTR exaptation to drive tissue-specific gene expression. PMID:21865161

  6. IL-2 suppression of IL-12p70 by a recombinant HSV-1 expressing IL-2 induces T-cell auto-reactivity and CNS demyelination.

    Directory of Open Access Journals (Sweden)

    Mandana Zandian

    Full Text Available To evaluate the role of cellular infiltrates in CNS demyelination in immunocompetent mice, we have used a model of multiple sclerosis (MS in which different strains of mice are infected with a recombinant HSV-1 expressing IL-2. Histologic examination of the mice infected with HSV-IL-2 demonstrates that natural killer cells, dendritic cells, B cells, and CD25 (IL-2rα do not play any role in the HSV-IL-2-induced demyelination. T cell depletion, T cell knockout and T cell adoptive transfer experiments suggest that both CD8(+ and CD4(+ T cells contribute to HSV-IL-2-induced CNS demyelination with CD8(+ T cells being the primary inducers. In the adoptive transfer studies, all of the transferred T cells irrespective of their CD25 status at the time of transfer were positive for expression of FoxP3 and depletion of FoxP3 blocked CNS demyelination by HSV-IL-2. The expression levels of IL-12p35 relative to IL-12p40 differed in BM-derived macrophages infected with HSV-IL-2 from those infected with wild-type HSV-1. HSV-IL-2-induced demyelination was blocked by injecting HSV-IL-2-infected mice with IL-12p70 DNA. This study demonstrates that suppression of the IL-12p70 function of macrophages by IL-2 causes T cells to become auto-aggressive. Interruption of this immunoregulatory axis results in demyelination of the optic nerve, the spinal cord and the brain by autoreactive T cells in the HSV-IL-2 mouse model of MS.

  7. Antitumor effect of murine dendritic and tumor cells transduced with IL-2 gene Antitumor effect of murine dendritic and tumor cells transduced with IL-2 gene

    Directory of Open Access Journals (Sweden)

    Justyna Wojas-Turek

    2012-10-01

    Full Text Available Interleukin (IL- 2 acts on a number of types of immune cells promoting their effector functions. To replace
    systemic administration of recombinant form of this cytokine, various genetically modified cells have been used in
    different preclinical models for tumor growth inhibition. In this study, dendritic or tumor cells transduced with retroviral
    vector carrying IL-2 gene (JAWS II/IL-2, X63/IL-2, MC38/IL-2 cells alone or combined with tumor antigenstimulated
    dendritic cells (JAWS II/TAg were exploited to treat colon carcinoma MC38-bearing mice. After the
    peritumoral injection of vaccine cells, the tumor growth delay and the increase in the number of tumor infiltrating
    CD4+ and CD8+ T lymphocytes were noted. A considerable increase in CD4+ cell influx into tumor tissue was observed
    when JAWS II/IL-2 cells or JAWS II/TAg with syngeneic MC38/IL-2 cells were applied. The increase in
    intensity of CD8+ cell infiltration was associated with immune reaction triggered by the same combination of applied
    cells or JAWS II/TAg with allogeneic X63/IL-2 cells. The effect observed in vivo was accompanied by MC38/0 cell
    specific cytotoxic activity of spleen cells in vitro. Thus, the application of vaccines, including IL-2-secreting cells of
    various origins, was able to induce different antitumor responses polarized by exogenous IL-2 and the encountered
    tumor antigen.Interleukin (IL- 2 acts on a number of types of immune cells promoting their effector functions. To replace
    systemic administration of recombinant form of this cytokine, various genetically modified cells have been used in
    different preclinical models for tumor growth inhibition. In this study, dendritic or tumor cells transduced with retroviral
    vector carrying IL-2 gene (JAWS II/IL-2, X63/IL-2, MC38/IL-2 cells alone or combined with tumor antigenstimulated
    dendritic cells (JAWS II/TAg were exploited to treat colon

  8. Costimulation of IL-2 Production through CD28 Is Dependent on the Size of Its Ligand.

    Science.gov (United States)

    Lim, Hong-Sheng; Cordoba, Shaun-Paul; Dushek, Omer; Goyette, Jesse; Taylor, Alison; Rudd, Christopher E; van der Merwe, P Anton

    2015-12-01

    Optimal T cell activation typically requires engagement of both the TCR and costimulatory receptors, such as CD28. Engagement of CD28 leads to tyrosine phosphorylation of its cytoplasmic region and recruitment of cytoplasmic signaling proteins. Although the exact mechanism of CD28 signal transduction is unknown, CD28 triggering has similarities to the TCR, which was proposed to use the kinetic-segregation (KS) mechanism. The KS model postulates that, when small receptors engage their ligands within areas of close (∼15 nm) contact in the T cell/APC interface, this facilitates phosphorylation by segregating the engaged receptor/ligand complex from receptor protein tyrosine phosphatases with large ectodomains, such as CD45. To test this hypothesis, we examined the effect of elongating the extracellular region of the CD28 ligand, CD80, on its ability to costimulate IL-2 production by primary T cells. CD80 elongation reduced its costimulatory effect without abrogating CD28 binding. Confocal microscopy revealed that elongated CD80 molecules were less well segregated from CD45 at the T cell/APC interface. T cells expressing CD28 harboring a key tyrosine-170 mutation were less sensitive to CD80 elongation. In summary, the effectiveness of CD28 costimulation is inversely proportional to the dimensions of the CD28-CD80 complex. Small CD28-CD80 complex dimensions are required for optimal costimulation by segregation from large inhibitory tyrosine phosphatases. These results demonstrate the importance of ligand dimensions for optimal costimulation of IL-2 production by T cells and suggest that the KS mechanism contributes to CD28 signaling.

  9. Radiation causes increased production and decreased utilization of IL-2 in human mononuclear cells

    International Nuclear Information System (INIS)

    The effects of radiation on the kinetics of Interleukin-2 (IL-2) production and utilization by mononuclear cells (MNCs) were studied. Mononuclear cells from normal, healthy individuals were subjected to various doses of radiation ranging from 0 to 2,000 rad and cultured in the presence of PHA. Supernatants from these cultures were harvested at various periods and their IL-2 contents determined by both the standard bioassay and ELISA. A radiation dose of 800 rad and higher had a marked effect on both IL-2 production and consumption. Although the supernatants from both the irradiated and non-irradiated MNCs contained maximal concentrations of IL-2 between 8 and 24 h of culture, the former had three times as much IL-2 as the latter. An increase in IL-2-mRNA levels was also noticed in irradiated, PHA-stimulated cells. Moreover, the supernatants from irradiated MNCs collected as late as 72 h after the initiation of culture contained more than 30% of the total IL-2 produced compared to less than 8% in supernatants from non-irradiated cells. Supernatants from non-irradiated cells incubated further with irradiated cells contained relatively higher quantities of IL-2 than those incubated continuously with non-irradiated cells. Supernatants from co-cultures of irradiated and non-irradiated MNCs contained less than expected amounts of IL-2 in two of the three subjects. Despite a difference in both the production and consumption of IL-2 between the irradiated and non-irradiated cells, there was no difference in their ability to generate IL-2 receptors. The results indicate that inactivation of radiosensitive suppressor T cells is associated with superinduction of IL-2 mRNA, increased production and decreased consumption of IL-2 by MNCs, thereby resulting in increased accumulation of IL-2

  10. SILAC-based quantification of changes in protein tyrosine phosphorylation induced by Interleukin-2 (IL-2) and IL-15 in T-lymphocytes

    DEFF Research Database (Denmark)

    Osinalde, Nerea; Sánchez-Quiles, Virginia; Akimov, Vyacheslav;

    2015-01-01

    This data article presents the first large-scale quantitative phosphoproteomics dataset generated to decipher the signaling networks initiated by IL-2 and IL-15 in T-lymphocytes. Data was collected by combining immunoprecipitation of tyrosine phosphorylated proteins and TiO2-based phosphopeptide....... The data supplied in this article is related to the research work entitled "Simultaneous dissection and comparison of IL-2 and IL-15 signaling pathways by global quantitative phosphoproteomics" [1]. All mass spectrometry data have been deposited in the ProteomeXchange with the identifier PXD001129....

  11. Dual anti-OX40/IL-2 therapy augments tumor immunotherapy via IL-2R-mediated regulation of OX40 expression.

    Directory of Open Access Journals (Sweden)

    William L Redmond

    Full Text Available The provision of T cell co-stimulation via members of the TNFR super-family, including OX40 (CD134 and 4-1BB (CD137, provides critical signals that promote T cell survival and differentiation. Recent studies have demonstrated that ligation of OX40 can augment T cell-mediated anti-tumor immunity in pre-clinical models and more importantly, OX40 agonists are under clinical development for cancer immunotherapy. OX40 is of particular interest as a therapeutic target as it is not expressed on naïve T cells but rather, is transiently up-regulated following TCR stimulation. Although TCR engagement is necessary for inducing OX40 expression, the downstream signals that regulate OX40 itself remain unclear. In this study, we demonstrate that OX40 expression is regulated through a TCR and common gamma chain cytokine-dependent signaling cascade that requires JAK3-mediated activation of the downstream transcription factors STAT3 and STAT5. Furthermore, combined treatment with an agonist anti-OX40 mAb and IL-2 augmented tumor immunotherapy against multiple tumor types. Dual therapy was also able to restore the function of anergic tumor-reactive CD8 T cells in mice with long-term well-established (>5 wks tumors, leading to increased survival of the tumor-bearing hosts. Together, these data reveal the ability of TCR/common gamma chain cytokine signaling to regulate OX40 expression and demonstrate a novel means of augmenting cancer immunotherapy by providing dual anti-OX40/common gamma chain cytokine-directed therapy.

  12. Analysis of IL-2-like factor in lymphocyte culture supernatant of olive flounder, Paralichthys oliveaceus

    Institute of Scientific and Technical Information of China (English)

    WU Riqin; ZHANG Peijun; LI Jun; XU Yongli

    2005-01-01

    To study immune mechanism of fish lymphocyte we performed a proliferation assay and ELISA using monoclonal antibody against human IL-2. The result showed that an interleukin-2 (IL-2)-like factor was detected in the supernatant of plant haemoglutinin (PHA)-stimulated lymphocyte culture from peripheral blood,spleen and head kidney of olive flounder, Paralichthys olivaceus. The quantities of IL-2-1ike factor in the supematant from different lymphoid tissues were quite different. The IL-2 like factor in the supernatant from cultured head kidney lymphocytes was much higher than those of peripheral blood lymphocytes and spleen lymphocytes (P<0.01). The IL-2 activity was found in either mouse thymocyte proliferation assay or flounder head kidney lymphocyte proliferation assay and shown to have obvious enhancing effect on proliferation of the above two types of cell. The recombinant human IL-2 (rhIL-2) was able to stimulate flounder thymocyte proliferation and used to detect the IL-2 receptor (IL-2R) on the surface of flounder lymphocyte. The cross-reaction between the lymphocytes of flounder peripheral blood and CD25(IL-2R) was detected with flow cytometry and shown that the percentage of CD25-positive cell in peripheral blood was 7.74± 0.67%.

  13. EFFECT OF MODIFIDE JADE SCREEN POWDER ON WOUNDED STRESS MICE IL-2 AND IL-2R GENIC EXPRESSION%加味玉屏风散对创伤应激小鼠IL-2IL-2R基因表达影响的形态计量学研究

    Institute of Scientific and Technical Information of China (English)

    曾广仙; 刘俊英

    2003-01-01

    目的:探讨加味玉屏风散对创伤应激小鼠脾淋巴细胞IL-2IL-2R基因表达的影响.方法:应用体外细胞培养和Northern印迹法图像分析仪定量检测了加味玉屏风散对刀豆蛋白A(C onA)诱导的小鼠脾淋巴细胞IL-2IL-2R基因表达的影响.结果:发现加味玉屏风散体内应用对创伤应激小鼠IL-2mRNA及IL-2RmRNA水平均具有明显的升高作用.结论:加味玉屏风散可通过促进创伤应激小鼠脾淋巴细胞IL-2IL-2R的基因转录表达进而逆转IL-2IL-2R的抑制状态.

  14. Alteration on the serum IL-2, IL-6 and receptors levels in patients with chronic glomerulonephritis

    International Nuclear Information System (INIS)

    Objective: To investigate the alteration and clinical significance of serum IL-2, IL-6 and receptors levels in Patients with Chronic Glomerulonephritis (CGN). Methods: Serum IL-2, IL-6 levels were determined by the bioassay methods, Serum sIL-2R, sIL-6R were determined by ELISA analysis, mIL-2R were determined by immunofluorescence assay in patient with CGN, and the data were compared with that in the normal controls group. Results: It was found that the levels of serum IL-6, sIL-2R and sIL-6R were significantly higher in CGN than that in controls (P < 0.0001), respectively. While serum IL-2 markedly lower in CGN than in the controls (P < 0.0001) and mIL-2R in CGN have no apparent difference with that in the controls. There was a positive correlation between IL-6 and BUN (P < 0.005), positive correlation between sIL6R and SCR. (P < 0.005), While there were positive correlation between sIL-2R and BUN. SCr(P < 0.05; P < 0.005) respectively, negative correlation between sIL-2R and urinary protein (P < 0.05). Conclusion: IL-2, sIL-2R, IL-6, sIL-6R play an important role in the pathogenesis of CGN, could reflect severity degree and prognosis of CGN. While low levels of IL-2 attributes to cell immune states of patients with CGN was depressed

  15. IL-2 and IL-15 Exhibit Opposing Effects on Fas Mediated Apoptosis

    Institute of Scientific and Technical Information of China (English)

    GulcinDemirci; XianChangLi

    2004-01-01

    It has been shown that IL-2 and IL-15 can have opposing effects on life and death of T cells. However, the roie of IL-2 and IL-15 in regulating the fate of other cell types is less clear. In the present study, we examined the impact of IL-2 and IL-15 on life and death of pre-B cells using the BAF-B03 line. We showed that BAF-B03 cells constitutively expressed the private IL-2Rα chain and IL-15Rα chain, and the shared IL-2Rβ chain and γc hain. Stimulation of BAF-B03 cells in vitro with IL-2 and IL-15 induced vigorous cell proliferation in a dose-dependent fashion. Titration of IL-2 and IL-15 in the assay showed that the mitotic effects of IL-2 and IL-15 were remarkably similar. However, the sensitivities of BAF-B03 cells to Fas mediated apoptosis after IL-2 and IL-15 stimulation were strikingly different. Cells cultured in IL-2 readily underwent apoptotic cell death upon cross-linking of the Fas receptor whereas cells cultured in IL-15 were extremely resistant to Fas triggered cell death. The anti-apoptotic effect of IL-15 in this model was associated with increased expression of Bcl-xL. FLIP expression, however, was comparable between IL-2 and IL-15 stimulated cells. We conclude that IL-2 and IL-15 have diametrically opposite effect on the fate of BAF-B03 cells, although both cytokines share similar receptor structure and exhibit similar mitotic activities. Cellular & Molecular Immunology. 2004;1(2):123-128.

  16. IL-2 and IL-15 Exhibit Opposing Effects on Fas Mediated Apoptosis

    Institute of Scientific and Technical Information of China (English)

    Gulcin Demirci; Xian Chang Li

    2004-01-01

    It has been shown that IL-2 and IL-15 can have opposing effects on life and death of T cells. However, the role of IL-2 and IL-15 in regulating the fate of other cell types is less clear. In the present study, we examined the impact of IL-2 and IL-15 on life and death of pre-B cells using the BAF-B03 line. We showed that BAF-B03 cells constitutively expressed the private IL-2Rα chain and IL-15Rα chain, and the shared IL-2Rβ chain and γc chain. Stimulation of BAF-B03 cells in vitro with IL-2 and IL-15 induced vigorous cell proliferation in a dose-dependent fashion. Titration of IL-2 and IL-15 in the assay showed that the mitotic effects of IL-2 and IL-15 were remarkably similar, Howerer, the sensitivities of BAF-B03 cells to Fas mediated apoptosis after IL-2 and IL-15 stimulation were strikingly different. Cells cultured in IL-2 readily underwent apoptotic cell deathupon cross-linking of the Fas receptor whereas cells cultured in IL-15 were extremely resistant to Fas triggered cell death. The anti-apoptotic effect of IL-15 in this model was associated with increased expression of Bcl-xL. FLIP expression, however, was comparable between IL-2 and IL-15 stimulated cells. We conclude that IL-2 and IL-15 have diametrically opposite effect on the fate of BAF-B03 cells, although both cytokines share similar receptor structure and exhibitsimilarmitoticactivities.

  17. On the Role of sIL-2R Measurements in Rheumatoid Arthritis and Cancers

    Directory of Open Access Journals (Sweden)

    Anna Maria Witkowska

    2005-01-01

    diseases of different pathology. Moreover, the soluble receptor has been considered, at least in part, responsible for unsuccessful immunotherapy with IL-2 in cancers. Several lines of evidence indicate sIL-2R measurements to be useful in determining disease progress and prognosis. This review summarizes current knowledge on the sIL-2R behavior in RA and solid cancers of varied etiology.

  18. Distinct Upstream Role of Type I IFN Signaling in Hematopoietic Stem Cell-Derived and Epithelial Resident Cells for Concerted Recruitment of Ly-6Chi Monocytes and NK Cells via CCL2-CCL3 Cascade.

    Directory of Open Access Journals (Sweden)

    Erdenebileg Uyangaa

    Full Text Available Type I interferon (IFN-I-dependent orchestrated mobilization of innate cells in inflamed tissues is believed to play a critical role in controlling replication and CNS-invasion of herpes simplex virus (HSV. However, the crucial regulators and cell populations that are affected by IFN-I to establish the early environment of innate cells in HSV-infected mucosal tissues are largely unknown. Here, we found that IFN-I signaling promoted the differentiation of CCL2-producing Ly-6Chi monocytes and IFN-γ/granzyme B-producing NK cells, whereas deficiency of IFN-I signaling induced Ly-6Clo monocytes producing CXCL1 and CXCL2. More interestingly, recruitment of Ly-6Chi monocytes preceded that of NK cells with the levels peaked at 24 h post-infection in IFN-I-dependent manner, which was kinetically associated with the CCL2-CCL3 cascade response. Early Ly-6Chi monocyte recruitment was governed by CCL2 produced from hematopoietic stem cell (HSC-derived leukocytes, whereas NK cell recruitment predominantly depended on CC chemokines produced by resident epithelial cells. Also, IFN-I signaling in HSC-derived leukocytes appeared to suppress Ly-6Ghi neutrophil recruitment to ameliorate immunopathology. Finally, tissue resident CD11bhiF4/80hi macrophages and CD11chiEpCAM+ dendritic cells appeared to produce initial CCL2 for migration-based self-amplification of early infiltrated Ly-6Chi monocytes upon stimulation by IFN-I produced from infected epithelial cells. Ultimately, these results decipher a detailed IFN-I-dependent pathway that establishes orchestrated mobilization of Ly-6Chi monocytes and NK cells through CCL2-CCL3 cascade response of HSC-derived leukocytes and epithelium-resident cells. Therefore, this cascade response of resident-to-hematopoietic-to-resident cells that drives cytokine-to-chemokine-to-cytokine production to recruit orchestrated innate cells is critical for attenuation of HSV replication in inflamed tissues.

  19. Phase I trial with recombinant interleukin-2 (rIL-2): immune activation by rIL-2 alone or following pretreatment with recombinant interferon-gamma.

    Science.gov (United States)

    Farace, F; Mathiot, C; Brandely, M; Tursz, T; Dorval, T; Pouillart, P; Triebel, F; Hercend, T; Fridman, W H

    1990-01-01

    Alterations of immunological parameters were analysed in patients with advanced malignancies during a phase I trial with rIL-2. Five-day infusions of rIL-2 at doses from 1 x 10(6) to 24 x 10(6) biological response modifiers program (BRMP) U/m2 per day were given to 29 patients, with a minimum of three patients per dose. The dose of 24 x 10(6) U/m2 per day was the maximal tolerated dose (MTD). Immunological parameters were analyzed at days 0, 8 and 11 of the rIL-2 courses. Following a leucopenia during rIL-2 infusion, a lymphocytosis was found in all patients except one. The lymphocytosis peaked at day 8 and was detected at doses of rIL-2 as low as 1 x 10(6) U/m2 per day, reaching a plateau at a dose of 16 x 10(6) U/m2 per day. Although all lymphocyte subsets were increased in patients receiving rIL-2, some patients had predominant T cells (CD3+, NKH1(CD56)-), others had predominant natural killer (NK) cells (CD3-, NKH1 (CD56)+), and yet others showed a mixed profile. A strong induction of cells cytotoxic for K562 targets was found in all patients at days 8 and 11. Eighteen patients received, 1 month later, a second treatment in which infusion of rIL-2 was preceded by a course of 5 days infusion of 2 x 10(6) U/m2 per day recombinant interferon-gamma (rIFN-gamma). The infusion of rIFN-gamma prior to rIL-2 had no effect on the rIL-2-induced alterations of immunological parameters. Taken together, our results suggest that immune stimulation by rIL-2 occurs even at low doses and is maximal at a dose below the MTD; and that pretreatment with low-dose rIFN-gamma does not modify the immune stimulation by rIL-2. PMID:2122928

  20. Analysis of IL-2-like factor in lymphocyte culture supernatant of olive flounder, Paralichthys oliveaceus

    Science.gov (United States)

    Wu, Riqin; Zhang, Peijun; Li, Jun; Xu, Yongli

    2005-03-01

    To study immune mechanism of fish lymphocyte we performed a proliferation assay and ELISA using monoclonal antibody against human IL-2. The result showed that an interleukin-2 (IL-2)-like factor was detected in the supernatant of plant haemoglutinin (PHA)-stimulated lymphocyte culture from peripheral blood, spleen and head kidney of olive flounder, Paralichthys olivaceus. The quantities of IL-2-like factor in the supernatant from different lymphoid tissues were quite different. The IL-2 like factor in the supernatant from cultured head kidney lymphocytes was much higher than those of peripheral blood lymphocytes and spleen lymphocytes ( Precombinant human IL-2, (rhIL-2) was able to stimulate flounder thymocyte proliferation and used to detect the IL-2 receptor (IL-2R) on the surface of flounder lymphocyte. The cross-reaction between the lymphocytes of flounder peripheral blood and CD25(IL-2R) was detected with flow cytometry and shown that the percentage of CD25-positive cell in peripheral blood was 7.74±0.67%.

  1. Interaction of interleukin-2 (IL-2) mutant proteins with interleukin-2 receptors

    International Nuclear Information System (INIS)

    The authors have previously produced several human IL-2 mutant proteins by site specific mutagenesis. Deletion or substitution of alanine for cysteine at positions 58 and 105 results in the decrease of biological activities. Substitution of serine for cysteine at position 125 does not affect the activity, however, deletion of this cysteine or amino acids in its vicinity causes a dramatic loss of activity. In this study, the interaction of these mutant proteins with IL-2 receptors has been analyzed by evaluating the competition between these mutant proteins and recombinant DNA derived IL-2 (rIL-2) for the binding to murine CTLL-2, an IL-2 dependent cell line. Addition of unlabeled rIL-2 (1 x 10-11 to 10-7M) inhibited the binding of I125-labeled rIL-2 (1 x 10-10M, specific activity 39.6 uCi/mg) to CTLL-2 cells in a concentration dependent manner. Mutant proteins with substitution of alanine for cysteine at position 58 (Ala 58) or deletion of cysteine at position 125 (Des-Cys 125) required a 100-fold higher concentration than rIL-2 to reach 50% inhibition. These results indicate that the decrease of biological activity in mutant proteins is partly, if not primarily, due to the attenuation in their abilities to bind IL-2 receptors

  2. In-vitro characterization of novel and functional regulatory SNPs in the promoter region of IL2 and IL2R alpha in a Gabonese population

    OpenAIRE

    Huang Xiangsheng; Kühne Vera; Kun Jürgen F J; Soboslay Peter T; Lell Bertrand; TP, Velavan

    2012-01-01

    Abstract Background The selection pressure imposed by the parasite has a functional consequence on the immune genes, leading to altered immune function in which regulatory T cells (Tregs) induced by parasites during infectious challenges modulate or thwart T effector cell mechanism. Methods We identified and investigated regulatory polymorphisms in the immune gene IL2 and its receptor IL2R alpha (also known as CD25) in Gabonese individuals exposed to plentiful parasitic infections. Results We...

  3. Rescue of an in vitro neuron phenotype identified in Niemann-Pick disease, type C1 induced pluripotent stem cell-derived neurons by modulating the WNT pathway and calcium signaling.

    Science.gov (United States)

    Efthymiou, Anastasia G; Steiner, Joe; Pavan, William J; Wincovitch, Stephen; Larson, Denise M; Porter, Forbes D; Rao, Mahendra S; Malik, Nasir

    2015-03-01

    Niemann-Pick disease, type C1 (NPC1) is a familial disorder that has devastating consequences on postnatal development with multisystem effects, including neurodegeneration. There is no Food and Drug Administration-approved treatment option for NPC1; however, several potentially therapeutic compounds have been identified in assays using yeast, rodent models, and NPC1 human fibroblasts. Although these discoveries were made in fibroblasts from NPC1 subjects and were in some instances validated in animal models of the disease, testing these drugs on a cell type more relevant for NPC1 neurological disease would greatly facilitate both study of the disease and identification of more relevant therapeutic compounds. Toward this goal, we have generated an induced pluripotent stem cell line from a subject homozygous for the most frequent NPC1 mutation (p.I1061T) and subsequently created a stable line of neural stem cells (NSCs). These NSCs were then used to create neurons as an appropriate disease model. NPC1 neurons display a premature cell death phenotype, and gene expression analysis of these cells suggests dysfunction of important signaling pathways, including calcium and WNT. The clear readout from these cells makes them ideal candidates for high-throughput screening and will be a valuable tool to better understand the development of NPC1 in neural cells, as well as to develop better therapeutic options for NPC1.

  4. Activation of human T cells by a tumor vaccine infected with recombinant Newcastle disease virus producing IL-2

    NARCIS (Netherlands)

    Janke, M.; Peeters, B.; Zhao, H.; Leeuw, O.; Moormann, R.J.M.; Arnold, A.; Ziouta, Y.; Fournier, P.; Schirrmacher, V.

    2008-01-01

    A new recombinant (rec) Newcastle disease virus (NDV) with incorporated human interleukin 2 (IL-2) as foreign therapeutic gene [rec(IL-2)] will be described. The foreign gene in rec(IL-2) did not affect the main features of NDV replication nor its tumor selectivity. Biologically active IL-2 was prod

  5. mIL-2R, T cell subsets & hepatitis C

    Institute of Scientific and Technical Information of China (English)

    Chao-Pin Li; Ke-Xia Wang; Jian Wang; Bo-Rong Pan

    2002-01-01

    AIM: To study the levels of membrane interleukin-2 receptor(mIL-2R ) and T cell subsets in peripheral bloodmononuclear cells (PBMC) from patients with hepatitis Cand their role in the pathogenesis of hepatitis C.METHODS: The levels of mlL-2R and T cells subsets in PBMCWere detected by biotin- streptstividin (BSA) technique beforeand after stimulation with PHA in 203 patients with hepatitis Cwith HCV-RNA( + ), anti-HCV( + ), anti-HCV(-).RESULTS: The total expressive levels of mlL-2R before andafter stimulation with PHA(0.03 ± 0.01, 0.03 ± 0.02, 0.04 ± 0.02, 0.36±0.03), and Tcell subsets in PBMC (0.62±0.06,0.37 ± 0.05, 0.35 ± 0.07) were all lower in patients withhepatitis C than those in normal controls (0.66 ± 0.07, 0.41± 0.06, 0.31 ± 0.05, P < 0.01 ). Among the patients, thelevels of mlL-2R were lower in silence than those in situationof PHA inducting (P< 0.01). However, the levels of mlL-2Rwere similar in acute hepatitis C to that in chronic hepatitis C(P>0.05). The levels of CD3+, CD4+, CD4 +/CD8+ Were lov erand CD8 + was higher in patients with acute and chronichepatitis C with anti-HCV( + ) than those in normal controls (0.62±0.06, 0.37±0.05, 0.35±0.07, 1.18±0.30, 0.61±0.07, 0.37±0.05, 1.39±0.33, 0.31±0.05, P<0.05-P<0.01).CONCLUSION: The cellular immunity is obviously changed inpatients with hepatitis C. The levels of mlL-2R end activationof T cells am closely associated with chronicity of hepatitis C.

  6. Interleukin 5, a T-cell-derived B-cell differentiation factor also induces cytotoxic T lymphocytes.

    OpenAIRE

    Takatsu, K; Kikuchi, Y; Takahashi, T.; Honjo, T; Matsumoto, M.; Harada, N.; Yamaguchi, N.; Tominaga, A

    1987-01-01

    We describe an interleukin, termed interleukin 5, that is the recombinant product previously referred to as T-cell-replacing factor (TRF), B-cell growth factor II (BCGF II), or killer-helper factor (KHF). TRF has been defined as a T-cell-derived lymphokine that acts on activated B cells as a B-cell differentiation factor. We have previously demonstrated that TRF is identical to BCGF II and induces expression of receptors for interleukin 2 (IL-2) on activated B cells. We also have reported tha...

  7. Cloning and Sequence Analysis of Gallus IL-2 Gene and Prediction of Protein Structure%鸡IL-2基因的克隆、序列分析及蛋白结构预测

    Institute of Scientific and Technical Information of China (English)

    谢昆; 蒋成砚; 胡俊杰; 全舒舟; 宋银银

    2011-01-01

    According to GenBank in the IL-2 cDNA sequences, using Premier 5.0 software design a pairs of specific primer, the local Dorking inject 100 μg/ml Con A, total RNA was extracted from feeding 24 hours lymphocyte from spleen, by RT-PCR cloning colony IL-2 cDNA fragment. The DNA fragments were analyzed use DNAStar software between different species identity for comparison. Sequence analysis showed that IL-2 gene has an open reading frames for 432 nucleotide acids, encoding 143 amino acids and most of signal peptide, compared with published on Genbank, the nucleotide identity was 98.8%、 31.2%、 28.2%、 27.3%、30.6%、 26.2%、 31.7%、 30.1%、 30.8%、 26.6%、 30.6%. compared with published on Genbank, the Amino acids was 95.8%, 7.6%、 7.6%、 9.7%、 7.6%、 6.9%、 10.4% 11.8%, 7.6%, 9.0%, 7.6%, 10.4% with cattle, horse, cat,sheep, bovine, canis, people, capra, mice, cow of IL-2 sequence comparison The gallus IL-2 gene was successfully cloned, there is a species-specific. And construct a based on further study the biological effects of IL-2 gene in particular the use of IL-2 enhance the immune effect of DNA vaccine.%根据GenBank中已发表的鸡白细胞介素2(IL-2)mRNA基因序列,利用Premier 5.0软件设计一对特异性引物,采用RT-PCR技术,以ConA刺激的鸡外周血淋巴细胞为材料,从总RNA中扩增出鸡IL-2基因.经琼脂糖凝胶电泳显示扩增片段约为432 bp,分离纯化片段,克隆入pMD-18T载体,转化DH5α感受态细胞,获得阳性重组质粒,经酶切鉴定,测序结果显示,该基因全长432 bp,含有一个432bp的开放阅读框,编码143个氨基酸.生物软件分析结果表明该序列与GenBank中发表的鸡的IL-2的核苷酸序列同源性为98.8%,与黄牛、马、鸡、绵羊、牛、犬、人、山羊、鼠、水牛的核苷酸同源性分别为31.2%、28.2%、273%、30.6%、26.2%、31.7%、30.1%、30.8%、26.6%、30.6%.与GenBank中发表的鸡IL-2编码

  8. Dynamic changes of IL-2, sIL-2R, IL-6 and IL-8 in children with mycoplasma pneumonia and their clinical significance%肺炎支原体肺炎IL-2、sIL-2R、IL-6、IL-8检测

    Institute of Scientific and Technical Information of China (English)

    刘文彬; 赵文利; 汤雪琴; 袁丽

    2012-01-01

    目的 探讨肺炎支原体肺炎(MPP)急性期和恢复期免疫功能动态变化及临床意义.方法 检测IL-2、sIL-2R、IL-6、IL-8均采用ELISA双抗体夹心法.结果 与对照组比较,MPP患儿急性期sIL-2R、IL-6,IL-8值均明显升高(P<0.01),IL-2值均明显下降(P<0.01),与急性期比较,MPP患儿恢复期IL-2值均明显升高(P<0.01),而sIL-2R、IL-6、IL-8值均明显下降(P<0.01);与对照组比较,MPP患儿恢复期IL-2、sIL-2R、IL-6、IL-8均无明显改变(P>0.05).结论 MPP患儿存在细胞免疫功能低下及紊乱,IL-2、sIL-2R、IL-6及IL-8值恢复较快,检测MPP患儿免疫功能,特别是IL-2、sIL-2R、IL-6和IL-8水平对疗效及预后的判定有重要价值.%Objective To observe the dynamic changes of IL-2, sIL-2R, IL-6 and IL-8 in children with mycoplasma pneumonia( MPP) and to study their clinical significance. Methods The serum levels of IL-2, sIL-2R, IL-6 and IL-8 in 40 children with MPP during the acute stage and convalescent stage were examined with double-antibody sandwich enzyme-linked immu-nosobent assay, and the results were compared with those in 30 normal children. Results The levels of slL-2R, IL-6 and IL-8 were higher and IL-2 was lower during acute stage than those in the normal subjects (P <0. 01). Compared those during the convalescent stage, the level of IL-2 was higher and sIL-2R, IL-6 and IL-8 were lower (P <0. 01) during the acute stage. There were not any significant differences of the levels of these four substances between the normal children and MPP children during the convalescent stage. Conclusion The cellular immune function of children with MPP is impaired and the changes of IL-2, sIL-2R, IL-6 and IL-8 can be used to evaluate the curative effects and prognosis of MPP in children.

  9. Astaxanthin, a Carotenoid, Stimulates Immune Responses by Enhancing IFN-γ and IL-2 Secretion in Primary Cultured Lymphocytes in Vitro and ex Vivo

    Directory of Open Access Journals (Sweden)

    Kuan-Hung Lin

    2015-12-01

    Full Text Available Astaxanthin, a potent antioxidant carotenoid, plays a major role in modulating the immune response. In this study, we examined the immunomodulatory effects of astaxanthin on cytokine production in primary cultured lymphocytes both in vitro and ex vivo. Direct administration of astaxanthin (70–300 nM did not produce cytotoxicity in lipopolysaccharide (LPS, 100 µg/ mL- or concanavalin A (Con A, 10 µg/ mL-activated lymphocytes, whereas astaxanthin alone at 300 nM induced proliferation of splenic lymphocytes (p < 0.05 in vitro. Although astaxanthin, alone or with Con A, had no apparent effect on interferon (INF-γ and interleukin (IL-2 production in primary cultured lymphocytes, it enhanced LPS-induced INF-γ production. In an ex vivo experiment, oral administration of astaxanthin (0.28, 1.4 and 7 mg/kg/day for 14 days did not cause alterations in the body or spleen weights of mice and also was not toxic to lymphocyte cells derived from the mice. Moreover, treatment with astaxanthin significantly increased LPS-induced lymphocyte proliferation ex vivo but not Con A-stimulated lymphocyte proliferation ex vivo. Enzyme linked immunosorbent assay (ELISA analysis revealed that administration of astaxanthin significantly enhanced INF-γ production in response to both LPS and Con A stimulation, whereas IL-2 production increased only in response to Con A stimulation. Also, astaxanthin treatment alone significantly increased IL-2 production in lymphocytes derived from mice, but did not significantly change production of INF-γ. These findings suggest that astaxanthin modulates lymphocytic immune responses in vitro, and that it partly exerts its ex vivo immunomodulatory effects by increasing INF-γ and IL-2 production without inducing cytotoxicity.

  10. Expression of low-, intermediate-, and high-affinity IL-2 receptors on B cell lines derived from patients with undifferentiated lymphoma of Burkitt's and non-Burkitt's types

    Energy Technology Data Exchange (ETDEWEB)

    Benjamin, D.; Rosolen, A.; Wormsley, S.B.; DeBault, L.E.; Colamonici, O.R. (Saint Francis Research Institute, Oklahoma City, OK (USA))

    1990-08-01

    IL-2 receptors on T cells exist in at least three forms which differ in their ligand-binding affinity. The low-affinity IL-2 receptor (IL-2R) consists of the 55-kDa Tac protein (p55 alpha), the intermediate-affinity site corresponds to the 70-kDa molecule (p70 beta), and the high-affinity IL-2R consists of a noncovalent heterodimeric structure involving both p55 alpha and p70 beta. We studied 24 B cell lines (8 EBV-negative and 16 EBV-positive) for IL-2R expression in the presence or absence of the tumor promoter, teleocidin. 125I-IL-2 radioreceptor binding assays and crosslinking studies demonstrated the sole expression of p55 alpha in EBV-negative cell lines only, whereas p55 alpha present in EBV-positive cell lines was always associated with p70 beta to construct high-affinity IL-2R. p70 beta was not detected in any of the EBV-negative cell lines, but was expressed on most of the EBV-positive cell lines (13 of 16). Our data also indicate that the expression of p55 alpha and p70 beta by radiolabeling correlates with their expression in flow cytometry, and that a large excess of p55 alpha is required to construct high-affinity IL-2R. Coexpression of p55 alpha and p70 beta on human B cells contributed to constructing high-affinity IL-2R hybrid complex as shown by rapid association rate contributed by p55 alpha and slow dissociation rate by p70 beta; teleocidin's ability to induce p55 alpha on cell lines which express p70 beta only, resulting in appearance of high-affinity IL-2R; and blocking p55 alpha by anti-Tac mAb in cell lines which constitutively express high-affinity IL-2R eliminated both high- and low-affinity components. The existence of low, intermediate, and high IL-2R on human B cells bears important future implications for understanding the mechanism of IL-2 signaling and the role of IL-2 in B cell activation, proliferation, and differentiation.

  11. IL-2 regulates SEB induced toxic shock syndrome in BALB/c mice.

    Directory of Open Access Journals (Sweden)

    Aslam Ali Khan

    Full Text Available BACKGROUND: Toxic Shock Syndrome (TSS is characterized by fever, rash, hypotension, constitutional symptoms, and multi-organ involvement and is caused by Staphylococcus aureus enterotoxins such as Staphylococcal Enterotoxin B (SEB. SEB binds to the MHC-IIalpha chain and is recognized by the TCRbeta chain of the Vbeta8 TCR(+ T cells. The binding of SEB to Vbeta chain results in rapid activation of T cells and production of inflammatory cytokines, such as Interleukin-2 (IL-2, Interferon-gamma and Tumor Necrosis Factor-alpha which mediate TSS. Although IL2 was originally identified as the T cell growth factor and was proposed to contribute to T cell differentiation, its role in TSS remains unexplored. METHODOLOGY/PRINCIPAL FINDINGS: Mice were injected with D-Gal (25 mg/mouse. One hour after D-Galactosamine (D-Gal injection each mouse was injected with SEB (20 microg/mouse. Mice were then observed for 72 hrs and death was recorded at different times. We tested Interleukin-12, IFNgamma, and IL-2 deficient mice (IL-2(-/-, but only the IL-2 deficient mice were resistant to SEB induced toxic shock syndrome. More importantly reconstitution of IL-2 in IL-2 deficient mice restored the shock. Interestingly, SEB induced IL-2 production from T cells was dependent on p38MAPK activation in macrophages as inhibition of it in macrophages significantly inhibited IL-2 production from T cells. CONCLUSION: This study shows the importance of IL -2 in TSS which has not been previously explored and it also shows that regulating macrophages function can regulate T cells and TSS.

  12. Effect of recombinant chicken IL-2 against chicken coccidiosis%重组鸡IL-2的抗球虫作用

    Institute of Scientific and Technical Information of China (English)

    谢昆; 李祥瑞

    2006-01-01

    鸡白细胞介素2(Ch IL-2)是一种重要的细胞因子,对Ch IL-2的抗球虫作用做了初步研究,分别在14日龄和21日龄给实验雏鸡胸部肌肉注射不同剂量的重组Ch IL-2蛋白,28日龄口服接种1×105个活球虫卵囊,结果表明,重组Ch IL-2蛋白能降低相对卵囊产量、减少盲肠病变、提高相对增重.其中以注射250μg重组Ch IL-2蛋白的试验组抗球虫效果最好,相对卵囊产量为23.26%,盲肠病变记分分别为0.4,相对增重率为97.80%,综合抗球虫指数(ACI)达到192.8.抗球虫效果良好,明显的优于其他试验组.

  13. IL-2 inducible T cell kinase (ITK) tunes T regulatory cell development and is required for suppressive function1

    OpenAIRE

    Huang, Weishan; Jeong, Ah-Reum; Kannan, Arun K.; Huang, Lu; August, Avery

    2014-01-01

    ITK is a key signaling mediator downstream of TcR, mediating T cell positive selection, innate T cell and CD4+ Th2/Th17 differentiation. Here we show that ITK also negatively tunes IL-2-induced expansion of Foxp3+ regulatory T cells (Treg). In vivo, Treg abundance is inversely correlated with ITK expression, and iTreg development is inversely dependent on ITK kinase activity. While Treg development normally requires both hematopoietic and thymic MHC class 2 (MHC2) expression, the absence of I...

  14. Inhibition of calcineurin abrogates while inhibition of mTOR promotes regulatory T cell expansion and graft-versus-host disease protection by IL-2 in allogeneic bone marrow transplantation.

    Directory of Open Access Journals (Sweden)

    Atsushi Satake

    Full Text Available Regulatory T cells (Tregs attenuate excessive immune responses, making their expansion beneficial in immune-mediated diseases including allogeneic bone marrow transplantation (BMT-associated graft-versus-host disease (GVHD. We have recently reported that Treg expansion does not require phospholipase Cγ activation when IL-2 is provided. As such, the combination of IL-2 and a calcineurin inhibitor (Cyclosporine A; CsA expands Tregs while inhibiting Tconv proliferation and protects against a mouse model of multiple sclerosis. However, CsA inhibits Treg proliferation in the presence of a TCR stimulus, suggesting that CsA may negatively impact Treg proliferation when they receive strong allogeneic MHC-mediated TCR signals. In this study, we show that CsA inhibits Treg proliferation and inducible Treg generation in allogeneic but not in syngeneic BMT when IL-2 is provided. In contrast to CsA, the mTOR inhibitor (Rapamycin almost completely suppressed IL-2-mediated Treg proliferation. However, CsA and Rapamycin inhibited Treg proliferation to a similar extent when TCR stimulation was provided. Furthermore, Rapamycin promoted Treg expansion and inducible Treg generation in allogeneic BMT recipients treated with IL-2. Consistent with these observations, CsA abrogated while Rapamycin promoted the protective effect of IL-2 on allogeneic BMT-induced GVHD. These results suggest that while CsA permits IL-2-induced Treg proliferation in the syngeneic setting (absence of strong TCR signals, CsA in combination with IL-2 may be detrimental for Treg proliferation in an allogeneic setting. Thus, in allogeneic settings, an mTOR inhibitor such as Rapamycin is a better choice for adjunct therapy with IL-2 in expansion of Tregs and protection against allogeneic BMT-induced GVHD.

  15. Low-Dose IL-2 Induces Regulatory T Cell-Mediated Control of Experimental Food Allergy.

    Science.gov (United States)

    Bonnet, Benjamin; Vigneron, James; Levacher, Béatrice; Vazquez, Thomas; Pitoiset, Fabien; Brimaud, Faustine; Churlaud, Guillaume; Klatzmann, David; Bellier, Bertrand

    2016-07-01

    Regulatory T cells (Tregs) are pivotal for maintenance of immune self-tolerance and also regulate immune responses to exogenous Ags, including allergens. Both decreased Treg number and function have been reported in allergic patients, offering new therapeutic perspectives. We previously demonstrated that Tregs can be selectively expanded and activated by low doses of IL-2 (ld-IL-2) inducing immunoregulation without immunosuppression and established its protective effect in autoimmune diseases. In this study, we evaluated the ability of ld-IL-2 to control allergy in an experimental model of food allergy. Ld-IL-2 induced Treg expansion and activation that elicited protection against clinical manifestations of food allergy in two mouse models with OVA and peanut. This clinical effect was lost in Treg-depleted mice, demonstrating the major contribution of Tregs in ld-IL-2 efficacy. Mechanistic studies further indicated that protection from allergy could be explained by a Treg-dependent local modification of the Th1/Th2 balance and an inhibition of mast cell recruitment and activation. Preventive and therapeutic effects of ld-IL-2 were observed over a 7-mo-period, highlighting its long-term efficacy. This study demonstrated that ld-IL-2 is efficient to prevent and to treat allergic immune responses, and thus represents a promising therapeutic strategy for managing allergic diseases. PMID:27259854

  16. mIL-2R, T cell subsets and hepatitis C

    OpenAIRE

    Li, Chao-Pin; Wang, Ke-Xia; Wang, Jian; Pan, Bo-Rong

    2002-01-01

    AIM: To study the levels of membrane interleukin-2 receptor (mIL-2R) and T cell subsets in peripheral blood mononuclear cells (PBMC) from patients with hepatitis C and their role in the pathogenesis of hepatitis C.

  17. Maintenance immunosuppression with intermittent intravenous IL-2 receptor antibody therapy in renal transplant recipients.

    LENUS (Irish Health Repository)

    Gabardi, Steven

    2011-09-01

    To report what we believe to be the first 2 cases of long-term (>24 months) intermittent intravenous interleukin-2 receptor antibody (IL-2RA) therapy for maintenance immunosuppression following renal transplantation.

  18. Intermediate- and high-affinity interleukin-2 receptors expressed in an IL-4-dependent T-cell line induce different signals.

    Science.gov (United States)

    Rebollo, A; Silva, A

    1993-01-01

    In order to understand the respective roles of IL-2R alpha and IL-2R beta subunits in transmission of the interleukin-2 (IL-2)-mediated growth signals, we have established two IL-4-dependent murine T-cell clones stably expressing the human IL-2R beta chain and three clones stably expressing the human IL-2R alpha chain. Whereas parental LD8 cells (which express only the murine IL-2R beta chain) do not proliferate in response to IL-2, cell lines stably expressing human IL-2R beta or the chimeric IL-2R alpha beta complex proliferate in response to IL-2. Stably transfected cells expressing the chimeric high-affinity receptor (human IL-2R alpha and murine IL-2R beta) expressed de novo endogenous murine IL-2R alpha when cultured in the presence of IL-2 but not IL-4. Both chimeric and endogenous receptors are functional in response to IL-2, since only addition of both anti-human and anti-murine IL-2R alpha monoclonal antibodies (mAb) inhibited IL-2-induced proliferation. Taken together, our results strongly suggest that human and murine IL-2R beta molecules are different since interaction of IL-2 with human p70 IL-2R is sufficient for transduction of proliferative signals in the absence of p55 IL-2R or, alternatively, that over-expression of the IL-2R beta chain renders cells responsive to IL-2. In addition, IL-2 stimulation of T cells through different forms of IL-2R results in the induction of distinct cellular responses. PMID:8262551

  19. Assay of sIL-2R and TNF-α in experimental allergic encephalomyelitis

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Objective: To investigate the change and effect of sIL-2R and TNF-α in the immunopathogenesis of experimental allergic encephalomyelitis(EAE). Methods: The EAE model was induced in guinea pigs. And the EAE animals were killed on the 8th, 15th and 22nd day after the MBP+CFA challenge. ConA-treated guinea pig spleen cells were cultured and supernatants were collected. The level of sIL-2R in supernatant was detected by ELISA, and the level of TNF-α in EAE was examined by biologic assay. Results: The EAE animals showed higher levels of sIL-2R and TNF-α than those of the normal control group. Conclusion: sIL-2R and TNF-α play an important role in the immunopathogenesis of EAE. This experiment offered thoretical evidence for further study of multiple sclerosis(MS) on pathogenesis and gave the clues for clinical use of immunospecific agents on MS.%目的:研究sIL-2R和TNF-α在实验性变态反应性脑脊髓炎(EAE)免疫发病机制中的变化与作用.方法:应用豚鼠诱导EAE动物模型.在MBP+CFA免疫豚鼠的第8、15、22天处死动物,取脾细胞,加入ConA诱生培养,收集上清液,采用ELISA方法测定sIL-2R水平,采用生物活性测定法检测TNF-α水平.结果:EAE组的sIL-2R与TNF-α水平明显高于正常对照组.结论:sIL-2R及TNF-α在EAE免疫发病机制中具有重要作用.

  20. EXPRESSION OF IL-2 AND SIL-2R AND ALTERATION OF CELL IMMUNITY IN PATIENTS WITH HYPERTENSIVE CEREBRAL HEMORRHAGE

    Institute of Scientific and Technical Information of China (English)

    张越林; 邱曙东; 师蔚; 党小军

    2006-01-01

    Interleukin-2(IL-2),an i mportant cytokinewith the other name as T cell growth factor,func-tions to promote the mitosis of hymphocytes,cani mprove the power of killer cells and help the pro-duction of antibody on the condition that it is com-bined with the specific hyperaviditic IL-2Rintargetcells.The t wo kinds of IL-2R,membrane boundIL-2R(mIL-2R)and soluble IL-2R(sIL-2R),com-pete with each other to combine with IL-2and thenbecome i mmunosuppressant[1].The i mportance oferythrocytes and T cells in the defe...

  1. Loss of immune tolerance to IL-2 in type 1 diabetes

    Science.gov (United States)

    Pérol, Louis; Lindner, John M.; Caudana, Pamela; Nunez, Nicolas Gonzalo; Baeyens, Audrey; Valle, Andrea; Sedlik, Christine; Loirat, Delphine; Boyer, Olivier; Créange, Alain; Cohen, José Laurent; Rogner, Ute Christine; Yamanouchi, Jun; Marchant, Martine; Leber, Xavier Charles; Scharenberg, Meike; Gagnerault, Marie-Claude; Mallone, Roberto; Battaglia, Manuela; Santamaria, Pere; Hartemann, Agnès; Traggiai, Elisabetta; Piaggio, Eliane

    2016-01-01

    Type 1 diabetes (T1D) is characterized by a chronic, progressive autoimmune attack against pancreas-specific antigens, effecting the destruction of insulin-producing β-cells. Here we show interleukin-2 (IL-2) is a non-pancreatic autoimmune target in T1D. Anti-IL-2 autoantibodies, as well as T cells specific for a single orthologous epitope of IL-2, are present in the peripheral blood of non-obese diabetic (NOD) mice and patients with T1D. In NOD mice, the generation of anti-IL-2 autoantibodies is genetically determined and their titre increases with age and disease onset. In T1D patients, circulating IgG memory B cells specific for IL-2 or insulin are present at similar frequencies. Anti-IL-2 autoantibodies cloned from T1D patients demonstrate clonality, a high degree of somatic hypermutation and nanomolar affinities, indicating a germinal centre origin and underscoring the synergy between cognate autoreactive T and B cells leading to defective immune tolerance. PMID:27708334

  2. Reliability of soluble IL-2 receptor measurements obtained with enzyme-linked immunosorbent assay

    International Nuclear Information System (INIS)

    Using an enzyme-linked immunosorbent assay (ELISA), human soluble interleukin-2 receptors (IL-2R) were measured in the serum of patients with various autoimmune system diseases. To study the sensitivity and specificity of the assay, soluble IL-2Rs were measured in the culture supernatants and in the cell extracts of peripheral blood mononuclear cells activated with phytohemagglutinin (PHA), purified protein derivative of tuberculin, and allogeneic lymphocytes, as well as in the serum of patients with various collagen diseases. The results correlated well with reports from other laboratories. For example, when stimulated by PHA, the greatest amount of soluble IL-2Rs was produced at the fastest rate. In addition, soluble IL-2R levels in the serum of collagen disease patients were significantly higher than those in healthy persons, who themselves exhibited low levels of detectable soluble IL-2Rs. It is hoped that reliable ELISA measurements of soluble IL-2Rs in the serum of atomic bomb survivors will assist in the interpretation of data collected during the work described in RP 2-87, a study of autoimmunity and autoimmune diseases in the Adult Health Study. (author)

  3. Endothelial cell-derived interleukin-6 regulates tumor growth

    International Nuclear Information System (INIS)

    Endothelial cells play a complex role in the pathobiology of cancer. This role is not limited to the making of blood vessels to allow for influx of oxygen and nutrients required for the high metabolic demands of tumor cells. Indeed, it has been recently shown that tumor-associated endothelial cells secrete molecules that enhance tumor cell survival and cancer stem cell self-renewal. The hypothesis underlying this work is that specific disruption of endothelial cell-initiated signaling inhibits tumor growth. Conditioned medium from primary human dermal microvascular endothelial cells (HDMEC) stably transduced with silencing RNA for IL-6 (or controls) was used to evaluate the role of endothelial-derived IL-6 on the activation of key signaling pathways in tumor cells. In addition, these endothelial cells were co-transplanted with tumor cells into immunodefficient mice to determine the impact of endothelial cell-derived IL-6 on tumor growth and angiogenesis. We observed that tumor cells adjacent to blood vessels show strong phosphorylation of STAT3, a key mediator of tumor progression. In search for a possible mechanism for the activation of the STAT3 signaling pathway, we observed that silencing interleukin (IL)-6 in tumor-associated endothelial cells inhibited STAT3 phosphorylation in tumor cells. Notably, tumors vascularized with IL-6-silenced endothelial cells showed lower intratumoral microvessel density, lower tumor cell proliferation, and slower growth than tumors vascularized with control endothelial cells. Collectively, these results demonstrate that IL-6 secreted by endothelial cells enhance tumor growth, and suggest that cancer patients might benefit from targeted approaches that block signaling events initiated by endothelial cells

  4. Effects of intermittent IL-2 alone or with peri-cycle antiretroviral therapy in early HIV infection: the STALWART study

    DEFF Research Database (Denmark)

    Tavel, Jorge A; Babiker, Abdel; Fox, Lawrence;

    2010-01-01

    BACKGROUND: The Study of Aldesleukin with and without antiretroviral therapy (STALWART) evaluated whether intermittent interleukin-2 (IL-2) alone or with antiretroviral therapy (ART) around IL-2 cycles increased CD4(+) counts compared to no therapy. METHODOLOGY: Participants not on continuous ART...... with > or = 300 CD4(+) cells/mm(3) were randomized to: no treatment; IL-2 for 5 consecutive days every 8 weeks for 3 cycles; or the same IL-2 regimen with 10 days of ART administered around each IL-2 cycle. CD4(+) counts, HIV RNA, and HIV progression events were collected monthly. PRINCIPAL FINDINGS: A total...

  5. Effects of intermittent IL-2 alone or with peri-cycle antiretroviral therapy in early HIV infection: the STALWART study

    OpenAIRE

    Tavel, Jorge A.; Abdel Babiker; Lawrence Fox; Daniela Gey; Gustavo Lopardo; Norman Markowitz; Nicholas Paton; Deborah Wentworth; Nicole Wyman

    2010-01-01

    BACKGROUND: The Study of Aldesleukin with and without antiretroviral therapy (STALWART) evaluated whether intermittent interleukin-2 (IL-2) alone or with antiretroviral therapy (ART) around IL-2 cycles increased CD4(+) counts compared to no therapy. METHODOLOGY: Participants not on continuous ART with > or = 300 CD4(+) cells/mm(3) were randomized to: no treatment; IL-2 for 5 consecutive days every 8 weeks for 3 cycles; or the same IL-2 regimen with 10 days of ART administered around each IL-2...

  6. Mannan-binding lectin inhibits proliferation of Jurkat cells and secretion of IL-2%MBL抑制Jurkat细胞增殖和分泌IL-2

    Institute of Scientific and Technical Information of China (English)

    王明永; 张雅妮; 雷鸣; 张丽芸; 卢晓; 陈政良

    2010-01-01

    甘露聚糖结合凝集素(MBL)在天然免疫中起关键作用,但其对T淋巴细胞介导获得性免疫应答是否有影响尚不得而知.用非特异性刺激物PHA、PMA及ionomycin活化人T淋巴细胞株Jurkat细胞,ELISA和流式细胞术检测MBL与Jur-kat细胞的结合能力,WST-1和ELISA分析MBL对PHA、PMA及ionomycin活化Jurkat细胞的增殖及分泌细胞因子IL-2的影响.结果显示:MBL可结合PHA/PMA活化的Jurkat细胞,与未活化Jurkat细胞结合微弱;高浓度MBL(10~20mg/L)能以浓度依赖的方式抑制PHA/PMA活化的Jurkat细胞增殖,加入外源IL-2不能逆转这种作用;高浓度MBL(10~20 mg/L)能抑制PMA/ionomycin活化的Jurkat细胞分泌细胞因子IL-2,且呈剂量依赖关系.提示MBL可能在调节T细胞介导的获得性免疫应答中起一定作用.

  7. A role for the intermediate affinity IL-2R in the protection against glucocorticoid-induced apoptosis.

    Science.gov (United States)

    Rebollo, A; Pitton, C; García, A; Gómez, J; Silva, A

    1995-01-01

    Recent work has shown that T lymphocytes undergo apoptosis upon treatment with the glucocorticoid analogue dexamethasone. These cells can be protected from the effect of dexamethasone by interleukin-2 (IL-2) or IL-4. We were interested in analysing whether a transfected cell dependent on three different lymphokines could be protected by them from the effect of dexamethasone. In addition, we took advantage of our cellular system, in which we expressed intermediate- or high-affinity IL-2R independently, to analyse the role of these receptors in the protection from glucocorticoid-induced apoptosis. In this report we show that IL-2 rescues murine T cells expressing exogenous intermediate- (TS1 beta) or high-affinity (TS1 alpha beta) IL-2 receptor (IL-2R) from dexamethasone-induced apoptosis. This result suggests that intermediate-affinity IL-2R alone can replace high-affinity IL-2R for the protection from the effect of dexamethasone. In addition, IL-4 and IL-9 are rescue-factors, as well as IL-2, of glucocorticoid-treated TS1 beta and TS1 alpha beta cells. Our data suggest that the presence of the alpha-chain of the IL-2R is not required for rescue by IL-2 from the effect of dexamethasone. In addition, we show that proliferation is not required for preventing glucocorticoid-induced apoptosis. This result implies a new role for the intermediate-affinity IL-2R. Images Figure 4 Figure 7 PMID:7751021

  8. Competition for IL-2 between regulatory and effector T cells to chisel immune responses

    Directory of Open Access Journals (Sweden)

    Thomas eHöfer

    2012-09-01

    Full Text Available In this review we discuss how the competition for cytokines between different cells of the immune system can shape the system wide immune response. We focus on interleukin-2 (IL-2 secretion by activated effector T cells (Teff and on the competition for IL-2 consumption between Teff and regulatory T cells (Treg. We discuss the evidence for the mechanism in which the depletion of IL-2 by Treg cells would be sufficient to suppress an autoimmune response, yet not strong enough to prevent an immune response. We present quantitative estimations and summarize our modeling effort to show that the tug-of-war between Treg and Teff cells for IL-2 molecules can be won by Treg cells in the case of weak activation of Teff leading to the suppression of the immune response. Or, for strongly activated Teff cells, it can be won by Teff cells bringing about the activation of the whole adaptive immune system. Finally, we discuss some recent applications attempting to achieve clinical effects through the modulation of IL-2 consumption by Treg compartment.

  9. Activated Human T Cells Secrete Exosomes That Participate in IL-2 Mediated Immune Response Signaling

    OpenAIRE

    Wahlgren, Jessica; Tanya De L Karlson; Glader, Pernilla; Telemo, Esbjörn; Valadi, Hadi

    2012-01-01

    It has previously been shown that nano-meter sized vesicles (30–100 nm), exosomes, secreted by antigen presenting cells can induce T cell responses thus showing the potential of exosomes to be used as immunological tools. Additionally, activated CD3+ T cells can secrete exosomes that have the ability to modulate different immunological responses. Here, we investigated what effects exosomes originating from activated CD3+ T cells have on resting CD3+ T cells by studying T cell proliferation, c...

  10. CLONING AND EXPRESSION OF THE GENE CODING FOR IL-2 (60)-PE40, A MOLECULAR TARGETED PROTEIN

    Institute of Scientific and Technical Information of China (English)

    张萌; 赵雪; 李焕娄; 卢圣栋

    1995-01-01

    It has recently been shown that chimeric toxin composed of IL2 fused tp PE40,a mutant form of Pseu-domonas Exotoxin A devoid of its native cell recognition and binding domain was cytotoxic to IL-2 receptor bearing cells.We here amplified the gene IL-2 (60),which codes for the N-terminal 1-60 amino acids of human IL-2 by PCR.After that,we fused it to PE40 and the new chimteric protein IL-2 (60)-PE40 was expressed in E.coli.SDS-PAGE revealed that IL-2(60)-PE40 chimeric protein accounts for more than 18% of total cell proteins.As the region IL-2 binds with its receptor was defined in the N-terminal residuse 8-54 of IL-2,such fusion proteins will have the same activity with IL-2-PE40.Following primany purification,IL-2(60)-PE40 was shown to be very toxic to IL-2 receptor-positive cells but non measurable effect on the cells lacking IL-2 receptors.Such a structure has not been reported by now.The fusion pro-tein is useful for suppressing the immune response in cases of rejection of allografts and organ transplants and as therapeutic agents for the treatment of IL-2 receptor related diseases such as autoimmune disease,and as therapeutic agents for the treatment of IL-2 receptor related diseases such as autoimmune disease,ATL(adult T-cell leukemia),et al.

  11. Genome wide mapping reveals PDE4B as an IL-2 induced STAT5 target gene in activated human PBMCs and lymphoid cancer cells.

    Directory of Open Access Journals (Sweden)

    Zsuzsanna S Nagy

    Full Text Available IL-2 is the primary growth factor for promoting survival and proliferation of activated T cells that occurs following engagement of the Janus Kinase (JAK1-3/and Signal Transducer and Activator of Transcription (STAT 5 signaling pathway. STAT5 has two isoforms: STAT5A and STAT5B (commonly referred to as STAT5 which, in T cells, play redundant roles transcribing cell cycle and survival genes. As such, inhibition of STAT5 by a variety of mechanisms can rapidly induce apoptosis in certain lymphoid tumor cells, suggesting that it and its target genes represent therapeutic targets to control certain lymphoid diseases. To search for these molecules we aligned IL-2 regulated genes detected by Affymetrix gene expression microarrays with the STAT5 cistrome identified by chip-on-ChIP analysis in an IL-2-dependent human leukemia cell line, Kit225. Select overlapping genes were then validated using qRT(2PCR medium-throughput arrays in human PHA-activated PBMCs. Of 19 putative genes, one key regulator of T cell receptor signaling, PDE4B, was identified as a novel target, which was readily up-regulated at the protein level (3 h in IL-2 stimulated, activated human PBMCs. Surprisingly, only purified CD8+ primary T-cells expressed PDE4B, but not CD4+ cells. Moreover, PDE4B was found to be highly expressed in CD4+ lymphoid cancer cells, which suggests that it may represent a physiological role unique to the CD8+ and lymphoid cancer cells and thus might represent a target for pharmaceutical intervention for certain lymphoid diseases.

  12. Potential role for IL-2 ELISpot in differentiating recent and remote infection in tuberculosis contact tracing.

    Directory of Open Access Journals (Sweden)

    Benjamin Krummel

    Full Text Available Interferon (IFN-gamma release assays (IGRA have improved tuberculosis contact tracing, but discrimination of recent from remote Mycobacterium tuberculosis contacts is not possible by IGRA alone. We present results of a tuberculosis contact investigation with a new early-secretory-antigenic-target (ESAT-6 and culture-filtrate-protein (CFP-10 specific interleukin (IL-2 ELISpot in addition to ESAT-6 and CFP-10 specific IFN-gamma ELISpot and tuberculin skin testing (TST. Results of the TST, IFN-gamma ELISpot and IL-2 ELISpot were positive in 6/172 (3.4%, 7/167 (4.2% and 6/196 (3.1% of contacts, respectively. Close contact (> or =100 hours to the index case increased the risk of positive results in the IFN-gamma ELISpot, TST, and IL-2 ELISpot by 40.8, 19.3, and 2.5 times, respectively. Individuals with a positive IFN-gamma ELISpot/negative IL-2 ELISpot result had a median (IQR duration of index case exposure of 568 hours (133_1000 compared to individuals with a positive IFN-gamma ELISpot/positive IL-2 ELISpot result (median = 24 hours; 20_130; p-value = 0.047. Combination of a M. tuberculosis specific IFN-gamma ELISpot with a M. tuberculosis specific IL-2 ELISpot significantly improved the identification of individuals with the highest risk of recent M. tuberculosis infection and is a promising method that should be explored to target tuberculosis preventive chemotherapy.

  13. Potential Role for IL-2 ELISpot in Differentiating Recent and Remote Infection in Tuberculosis Contact Tracing

    Science.gov (United States)

    Ernst, Martin; Reiling, Norbert; Eker, Barbara; Rath, Heidrun; Hoerster, Robert; Wappler, Waltraud; Glaewe, Andrea; Schoellhorn, Volker; Sotgiu, Giovanni; Lange, Christoph

    2010-01-01

    Interferon (IFN)-γ release assays (IGRA) have improved tuberculosis contact tracing, but discrimination of recent from remote Mycobacterium tuberculosis contacts is not possible by IGRA alone. We present results of a tuberculosis contact investigation with a new early-secretory-antigenic-target (ESAT)-6 and culture-filtrate-protein (CFP)-10 specific interleukin (IL)-2 ELISpot in addition to ESAT-6 and CFP-10 specific IFN-γ ELISpot and tuberculin skin testing (TST). Results of the TST, IFN-γ ELISpot and IL-2 ELISpot were positive in 6/172 (3.4%), 7/167 (4.2%) and 6/196 (3.1%) of contacts, respectively. Close contact (≥100 hours) to the index case increased the risk of positive results in the IFN-γ ELISpot, TST, and IL-2 ELISpot by 40.8, 19.3, and 2.5 times, respectively. Individuals with a positive IFN-γ ELISpot/negative IL-2 ELISpot result had a median (IQR) duration of index case exposure of 568 hours (133_1000) compared to individuals with a positive IFN-γ ELISpot/positive IL-2 ELISpot result (median = 24 hours; 20_130; p-value = 0.047). Combination of a M. tuberculosis specific IFN-γ ELISpot with a M. tuberculosis specific IL-2 ELISpot significantly improved the identification of individuals with the highest risk of recent M. tuberculosis infection and is a promising method that should be explored to target tuberculosis preventive chemotherapy. PMID:20652022

  14. Enriched retinal ganglion cells derived from human embryonic stem cells.

    Science.gov (United States)

    Gill, Katherine P; Hung, Sandy S C; Sharov, Alexei; Lo, Camden Y; Needham, Karina; Lidgerwood, Grace E; Jackson, Stacey; Crombie, Duncan E; Nayagam, Bryony A; Cook, Anthony L; Hewitt, Alex W; Pébay, Alice; Wong, Raymond C B

    2016-01-01

    Optic neuropathies are characterised by a loss of retinal ganglion cells (RGCs) that lead to vision impairment. Development of cell therapy requires a better understanding of the signals that direct stem cells into RGCs. Human embryonic stem cells (hESCs) represent an unlimited cellular source for generation of human RGCs in vitro. In this study, we present a 45-day protocol that utilises magnetic activated cell sorting to generate enriched population of RGCs via stepwise retinal differentiation using hESCs. We performed an extensive characterization of these stem cell-derived RGCs by examining the gene and protein expressions of a panel of neural/RGC markers. Furthermore, whole transcriptome analysis demonstrated similarity of the hESC-derived RGCs to human adult RGCs. The enriched hESC-RGCs possess long axons, functional electrophysiological profiles and axonal transport of mitochondria, suggestive of maturity. In summary, this RGC differentiation protocol can generate an enriched population of functional RGCs from hESCs, allowing future studies on disease modeling of optic neuropathies and development of cell therapies. PMID:27506453

  15. Fever and the use of paracetamol during IL-2-based immunotherapy in metastatic melanoma

    DEFF Research Database (Denmark)

    Køstner, Anne Helene; Ellegaard, Mai-Britt Bjørklund; Christensen, Ib Jarle;

    2015-01-01

    effective antitumor immune response. The purpose of this retrospective study was to examine the potential role of the IL-2-induced fever in melanoma patients treated with or without paracetamol in two consecutive cohorts. One hundred and seventy-nine patients with metastatic melanoma treated with a modified...... decrescendo regimen of IL-2 and Interferon (IFN) between 2004 and 2010 were retrospectively studied. 87 patients treated before 2007 received paracetamol as part of the treatment schedule, and 92 patients treated after 2007 did not receive paracetamol routinely. Body temperature was analyzed as dichotomized...

  16. Graves'眼病患者血清IL-2、sIL-2R和TGF-β测定的临床意义%Clinical Significance of Determination of Changes of Serum IL-2,sIL-2R and TGF-β Levels in Patients with Graves' Ophthalmopathy

    Institute of Scientific and Technical Information of China (English)

    李函

    2009-01-01

    目的:探讨Craves'眼病患者血清IL-2、sIL-2R和TGF-β水平的变化、病情进展和病因学的关系.方法:对照组30名和Craves眼病30例患者的血清TGF-β含量均采用放射免疫分析;血清IL-2、sIL-2R均采用酶联免疫吸附试验.结果:30例Graves'眼病患者血清IL-2治疗前水平非常显著低于对照组(P<0.01);经免疫抑制剂治疗6个月后水平较治疗前升高非常显著,与对照组比较已无显著性差异(P>0.05);sIL-2R含量治疗前非常显著的高于治疗后组和对照组(P<0.01);经用上述方案治疗后与对照组比较下降非常显著,但仍存在显著性差异(P<0.05);血清TGF-β浓度在治疗前非常显著地高于对照组(P<0.01);经治疗后显著下降,但与对照组比较差异仍非常显著(P<0.01).结论:Graves'眼病患者血清IL-2、sIL-2R和TGF-β三项血清指标的测定对于了解和认识其发病机理及预估病情有帮助.

  17. Effects of intermittent IL-2 alone or with peri-cycle antiretroviral therapy in early HIV infection: the STALWART study.

    Directory of Open Access Journals (Sweden)

    Jorge A Tavel

    Full Text Available BACKGROUND: The Study of Aldesleukin with and without antiretroviral therapy (STALWART evaluated whether intermittent interleukin-2 (IL-2 alone or with antiretroviral therapy (ART around IL-2 cycles increased CD4(+ counts compared to no therapy. METHODOLOGY: Participants not on continuous ART with > or = 300 CD4(+ cells/mm(3 were randomized to: no treatment; IL-2 for 5 consecutive days every 8 weeks for 3 cycles; or the same IL-2 regimen with 10 days of ART administered around each IL-2 cycle. CD4(+ counts, HIV RNA, and HIV progression events were collected monthly. PRINCIPAL FINDINGS: A total of 267 participants were randomized. At week 32, the mean CD4(+ count was 134 cells greater in the IL-2 alone group (p<0.001, and 133 cells greater in the IL-2 plus ART group (p<0.001 compared to the no therapy group. Twelve participants in the IL-2 groups compared to 1 participant in the group assigned to no therapy experienced an opportunistic event or died (HR 5.84, CI: 0.59 to 43.57; p = 0.009. CONCLUSIONS: IL-2 alone or with peri-cycle HAART increases CD4(+ counts but was associated with a greater number of opportunistic events or deaths compared to no therapy. These results call into question the immunoprotective significance of IL-2-induced CD4(+ cells. TRIAL REGISTRATION: ClinicalTrials.gov NCT00110812.

  18. Towards the Maturation and Characterization of Smooth Muscle Cells Derived from Human Embryonic Stem Cells

    OpenAIRE

    Helena Vazão; Ricardo Pires das Neves; Mário Grãos; Lino Ferreira

    2011-01-01

    In this study we demonstrate that CD34(+) cells derived from human embryonic stem cells (hESCs) have higher smooth muscle cell (SMC) potential than CD34(-) cells. We report that from all inductive signals tested, retinoic acid (RA) and platelet derived growth factor (PDGF(BB)) are the most effective agents in guiding the differentiation of CD34(+) cells into smooth muscle progenitor cells (SMPCs) characterized by the expression of SMC genes and proteins, secretion of SMC-related cytokines, co...

  19. Cell surface-expressed moesin-like receptor regulates T cell interactions with tissue components and binds an adhesion-modulating IL-2 peptide generated by elastase.

    Science.gov (United States)

    Ariel, A; Hershkoviz, R; Altbaum-Weiss, I; Ganor, S; Lider, O

    2001-03-01

    The adhesion of leukocytes to the extracellular matrix (ECM) depends on their responses to variations in the chemotactic signals in their milieu, as well as on the functioning of cytoskeletal and context-specific receptors. Ezrin, radixin, and moesin constitute a family of proteins that link the plasma membrane to the actin cytoskeleton. The surface expression of moesin on T cells and its role in cell adhesion has not been fully elucidated. Recently, we found that IL-2 peptides generated by elastase modified the adhesion of activated T cells to ECM ligands. Here, we further examined the adhesion regulatory effects of EFLNRWIT, one of the IL-2 peptides, as well as the existence and putative function of its receptor on T cells. We found that when presented to T cells in the absence of another activator, the EFLNRWIT peptide induced cell adhesion to vessel wall and ECM components. Binding of a radiolabeled peptide to T cells, precipitation with the immobilized peptide, and amino acid sequencing of the precipitated protein revealed that EFLNRWIT exerts its function via a cell surface-expressed moesin-like moiety, whose constitutive expression on T cells was increased after activation. This notion was further supported by our findings that: 1) anti-moesin mAb inhibited the binding of T cells to the immobilized EFLNRWIT peptide, 2) immobilized recombinant moesin bound the IL-2 peptide, and 3) soluble moesin inhibited the EFLNRWIT-induced T cell adhesion to fibronectin. Interestingly, moesin appears to be generally involved in T cell responses to adhesion-regulating signals. Thus, the IL-2 peptide EFLNRWIT appears to exert its modulating capacities via an adhesion-regulating moesin-like receptor. PMID:11207255

  20. Modeling human liver biology using stem cell-derived hepatocytes

    OpenAIRE

    Sun, Pingnan; Zhou, XiaoLing; Farnworth, Sarah; Arvind H Patel; Hay, David C.

    2013-01-01

    Stem cell-derived hepatocytes represent promising models to study human liver biology and disease. This concise review discusses the recent progresses in the field, with a focus on human liver disease, drug metabolism and virus infection.

  1. Modeling Human Liver Biology Using Stem Cell-Derived Hepatocytes

    OpenAIRE

    Arvind H Patel; Hay, David C.; Farnworth, Sarah L.; Pingnan Sun; Xiaoling Zhou

    2013-01-01

    Stem cell-derived hepatocytes represent promising models to study human liver biology and disease. This concise review discusses the recent progresses in the field, with a focus on human liver disease, drug metabolism and virus infection.

  2. Treatment with IL-2 and IL-12 inhibits tumour cell division in SL2 lymphoma

    NARCIS (Netherlands)

    Masztalerz, A; Van Luyn, M; Werner, N; Molema, G; Everse, LA; Den Otter, W

    2004-01-01

    We examined which mechanism plays a dominant role in the rejection of solid SL2 lymphoma treated with locally applied IL-2 and /or IL-12. This treatment resulted in about 80% cures. There was a moderate influx of leukocytes in the tissue surrounding tumours; yet these cells failed to invade the soli

  3. Expression of IL-2 and IL-11 and its significance in patients with ankylosing spondylitis

    Institute of Scientific and Technical Information of China (English)

    Feng Liu; Fan Wang; Cong-Cong Wang; Nuo Li; Shu-Feng Li

    2013-01-01

    Objective: To explore the expression of IL-2 and IL-11 and its significance in patients with ankylosing spondylitis (AS). Methods: A total of 48 active AS patients in our hospital and 40 normal control subjects were selected in our study. Bath ankylosing spondylitis disease activity index (BASDAI), Bath ankylosing spondylitis functional index (BASFI), Bath ankylosing spondylitis metrology index (BASMI), ESR and CRP expression levels were compared before treatment, 12 h after treatment and 24 h after treatment. IL-2 and IL-11 expression were also compared between these two groups. Results: The BASDAI score, BASFI score and BASMI score of the AS patients before treatment significantly decreased compared with those 12 weeks and 24 weeks after treatment (P0.05). Pearson's linear-correlation analysis showed that serum IL-2 level had a positive correlation with BASDAI, BASFI, BASMI, ESR and CRP (r=0.661,0.547,0.474,0.362,0.416, P<0.05) and serum IL-11 level had a negative correlation with BASDAI, BASFI, BASMI, ESR and CRP (r=-0.629,-0.412, -0.422, -0.387, -0.408, -0.315, P<0.05). Conclusions: Serum levels of IL-2 in active AS patients significantly increase and will decrease after treatment. However, serum levels of IL-11 significantly decrease and will increase after treatment, which indicates that serum IL-2 has a positive correlation with the degree of AS and serum IL-11 has a negative correlation with the degree of AS, both of which are correlated closely with the onset of AS.

  4. A self-inactivating retrovector incorporating the IL-2 promoter for activation-induced transgene expression in genetically engineered T-cells

    Directory of Open Access Journals (Sweden)

    Lejeune Laurence

    2006-11-01

    Full Text Available Abstract Background T-cell activation leads to signaling pathways that ultimately result in induction of gene transcription from the interleukin-2 (IL-2 promoter. We hypothesized that the IL-2 promoter or its synthetic derivatives can lead to T-cell specific, activation-induced transgene expression. Our objective was to develop a retroviral vector for stable and activation-induced transgene expression in T-lymphocytes. Results First, we compared the transcriptional potency of the full-length IL-2 promoter with that of a synthetic promoter composed of 3 repeats of the Nuclear Factor of Activated T-Cells (NFAT element following activation of transfected Jurkat T-cells expressing the large SV40 T antigen (Jurkat TAg. Although the NFAT3 promoter resulted in a stronger induction of luciferase reporter expression post stimulation, the basal levels of the IL-2 promoter-driven reporter expression were much lower indicating that the IL-2 promoter can serve as a more stringent activation-dependent promoter in T-cells. Based on this data, we generated a self-inactivating retroviral vector with the full-length human IL-2 promoter, namely SINIL-2pr that incorporated the enhanced green fluorescent protein (EGFP fused to herpes simplex virus thymidine kinase as a reporter/suicide "bifunctional" gene. Subsequently, Vesicular Stomatitis Virus-G Protein pseudotyped retroparticles were generated for SINIL-2pr and used to transduce the Jurkat T-cell line and the ZAP-70-deficient P116 cell line. Flow cytometry analysis showed that EGFP expression was markedly enhanced post co-stimulation of the gene-modified cells with 1 μM ionomycin and 10 ng/ml phorbol 12-myristate 13-acetate (PMA. This activation-induced expression was abrogated when the cells were pretreated with 300 nM cyclosporin A. Conclusion These results demonstrate that the SINIL-2pr retrovector leads to activation-inducible transgene expression in Jurkat T-cell lines. We propose that this design can be

  5. A study on recombinant human interleukin 2 radioiodinated with 125I (125I-rIL-2) for RIA

    International Nuclear Information System (INIS)

    RIL-2 was labelled with 125I by Iodogen method. The products were purified by HPLC. Analysis of each radiolabelled preparation showed that greater than 95% of the radioiodine was associated with a single protein peak. The specific activity of radioiodinated rIL-2 was approximately 2.2 x 1016-2.8 x 1016 Bq/mol. The 125I-rIL-2 was designed for quantitating human IL-2 with sensitive radioimmunoassay (RIA). The bioactivity of 125I-rIL-2 is good

  6. Long-term effects of intermittent IL-2 in HIV infection: extended follow-up of the INSIGHT STALWART Study

    OpenAIRE

    Markowitz, Norman; Lopardo, Gustavo; Wentworth, Deborah; Gey, Daniela; Babiker, Abdel; Fox, Lawrence; Tavel, Jorge; Fisher, Martin; for the STALWART Study Group,

    2012-01-01

    BACKGROUND The Study of Aldesleukin with and without Antiretroviral Therapy (STALWART) was designed to evaluate whether intermittent IL-2 alone or with peri-cycle ART increased CD4+ cell counts (and so delayed initiation of ART) in HIV infected individuals having ≥ 300 CD4+ cells/mm(3) compared to untreated controls. When the results of two large clinical trials, ESPRIT and SILCAAT, showed no clinical benefit from IL-2 therapy, IL-2 administration was halted in STALWART. Because IL-2 reci...

  7. Long-Term Effects of Intermittent IL-2 in HIV Infection: Extended Follow-Up of the INSIGHT STALWART Study

    OpenAIRE

    Markowitz, Norman; Lopardo, Gustavo; Wentworth, Deborah; Gey, Daniela; Babiker, Abdel; Fox, Lawrence; Tavel, Jorge; ,

    2012-01-01

    Background The Study of Aldesleukin with and without Antiretroviral Therapy (STALWART) was designed to evaluate whether intermittent IL-2 alone or with peri-cycle ART increased CD4+ cell counts (and so delayed initiation of ART) in HIV infected individuals having ≥300 CD4+ cells/mm3 compared to untreated controls. When the results of two large clinical trials, ESPRIT and SILCAAT, showed no clinical benefit from IL-2 therapy, IL-2 administration was halted in STALWART. Because IL-2 recipients ...

  8. Role of the multichain IL-2 receptor complex in the control of normal and malignant T-cell proliferation

    Energy Technology Data Exchange (ETDEWEB)

    Waldmann, T.A.

    1987-11-01

    Antigen-induced activation of resting T-cells induces the synthesis of interleukin-2 (IL-2), as well as the expression of specific cell surface receptors for this lymphokine. There are at least two forms of the cellular receptors for IL-2, one with a very high affinity and the other with a lower affinity. The authors have identified two IL-2 binding peptides, a 55-kd peptide reactive with the anti-Tac monoclonal antibody, and a novel 75-kd non-Tac IL-2 binding peptide. Cell lines bearing either the p55, Tac, or the p75 peptide along manifested low-affinity IL-2 binding, whereas cell lines bearing both peptides manifested both high- and low-affinity receptors. Fusion of cell membranes from low-affinity IL-2 binding cells bearing the Tac peptide alone with membranes from a cell line bearing the p75 peptide alone generates hybrid membranes bearing high-affinity receptors. They propose a multichain model for the high-affinity IL-2 receptor in which both the Tac and the p75 IL-2 binding peptides are associated in a receptor complex. In contrast to resting T-cells, human T-cell lymphotropic virus I-associated adult T-cell leukemia cells constitutively express large numbers of IL-2 receptors. Because IL-2 receptors are present on the malignant T-cells but not on normal resting cells, clinical trials have been initiated in which patients with adult T-cell leukemia are being treated with either unmodified or toxin-conjugated forms of anti-Tac monoclonal antibody directed toward this growth factor receptor. Cross-linking studies were done using (/sup 125/I) IL-2.

  9. SDF-1α通过 p38MAPK-HIF-1α通路加强卵巢癌细胞转移的机制探讨%Stromal cell-derived factor-1 alpha(SDF-1α) enhances cells invasion by p38MAPK-HIF-1αmediated signaling in ovarian cancer

    Institute of Scientific and Technical Information of China (English)

    薛宝瑶; 赵静; 张西愿; 姜霞; 周卓; 于月成

    2015-01-01

    目的:明确缺氧诱导因子-lα( HIF-1α)在基质细胞衍生因子-1α( SDF-1α)调控的卵巢癌细胞迁移中的作用及其机制。方法采用免疫组化分析HIF-1α在卵巢癌组织中的表达水平;将HIF-1αsiRNA转染SKOV3卵巢癌细胞,分别用SDF-1α和SDF-1α+SB203580处理,用RT-PCR及western blotting分析HIF-1α、p38MAPK的表达水平。结果免疫组化分析表明,HIF-1α在卵巢癌组织中高表达,同时SDF-1α可明显诱导HIF-1α的mRNA及蛋白水平的表达;当用HIF-1αsiRNA转染后,细胞中HIF-1α的表达水平明显下降。当用通路抑制剂SB203580处理后,SDF-1α诱导的HIF-1α水平明显降低。结论 SDF-1α可通过p38MAPK-HIF-1α通路来调控卵巢癌细胞的迁移。因此,该研究将为卵巢癌的防治提供新的研究方向。%Objective To observe the role and mechanism of hypoxia inducible factor-1α( HIF-1α) in ovarian cancer cell migration regulated by stromal cell-derived factor-1 alpha ( SDF-1α) .Methods The expression of HIF-1αin cervical cancer was analyzed by immunohistochemistry.HIF-1αsiRNA was transfected into ovarian cancer cell line SKOV3, and then treated with SDF-1αand SDF-1α+SB203580, respectively.The expression levels of HIF-1αand p38MAPK were confirmed by RT-PCR and western blotting.Results Immunohistochemical analysis showed that the expression of HIF-1αin ovarian cancer tissues was overexpressed, and that at the same time SDF-1αcould induce the expression of mRNA and protein levels in HIF-1α.After transfected by HIF-1αsiRNA, the HIF-1αexpression level decreased obviously.When treated by SB203580, the expression level of HIF-1αinduced by SDF-1αdecreased significantly. Conclusion SDF-1αcan regulate cells invasion by p38MAPK-HIF-1αmediated signaling in ovarian cancer.Therefore, this study will provide a new research direction for prevention and treatment of ovarian cancer.

  10. Evaluation in mice of the capillary leak syndrome (CLS) mediated by the systemic administration of recombinant interleukin-2 (IL-2)

    International Nuclear Information System (INIS)

    Since a CLS with interstitial pulmonary edema has been the major toxicity of IL-2 administration in humans, the authors studied this CLS in mice by quantitating the IL-2 mediated, tissue extravasation (Ex) of intravenously injected 125I-bovine serum albumin (BSA). Mice received saline (HBSS) or 200,000 U of IL-2 intraperitoneally thrice daily from day 0-6 before tissues were counted. A permeability index (PI) was calculated by dividing the mean counts per minute (CPM) from tissues of treated mice by those from controls. In a representative experiment, increased BSA Ex was noted in the lungs of mice treated with IL-2 when compared with HBSS (6187 +/- 141 and 638 +/- 64 CPM +/- SEM, respectively; p2 < .001; PI = 9.7). Other tissues with increased BSA Ex included the liver, spleen, kidneys and mesenteric lymph nodes (PI = 6.7, 10.0, 6.3, 6.0, respectively). BSA Ex, which did not occur with the excipient control, was dependent upon the dose and the duration of IL-2. Serial lung weights showed dramatic increases in water weight induced by IL-2. Treatment of mice with radiation (500R), cyclophosphamide, or cortisone acetate significantly reduced IL-2 mediated BSA Ex. Thus, IL-2 induced a generalized CLS which is mediated directly or indirectly by cellular mechanisms

  11. Serum concentration of IL-6, IL-2, TNF-α, and IFNγ in Vitiligo patients

    Directory of Open Access Journals (Sweden)

    Suman Singh

    2012-01-01

    Full Text Available Background: Vitiligo is an acquired depigmenting disorder characterized by the loss of functional melanocytes from the epidermis. Although the etiology of vitiligo is unknown, over the last few years, substantial data from clinical research has greatly supported the ′Autoimmune theory′ and this is supported by the frequent association of vitiligo with disorders that have an autoimmune origin, including Hashimoto′s thyroiditis, Graves disease, type 1 insulin-dependent diabetes mellitus, and Addison′s disease. As cytokines are important mediators of immunity, there is evidence to suggest that they play a major role in the pathogenesis of autoimmune diseases. Aim: Keeping this in view we have assayed sera for cytokine IL-6, IL-2, Tumor necrosis factor (TNF-α, and IFNγ in 80 cases of vitiligo and compared it with healthy subjects, in order to find out whether they play a role in the pathogenesis of vitiligo or not. Materials and Methods: Serum IL-6, IL-2, TNF-α, and IFNγ were done by the indirect enzyme linked immunosorbent assay (ELISA. Results: The mean serum IL-6 and IL-2 levels in the patient group were significantly higher when compared with those of the normal controls. The mean serum IFNγ level in patients with vitiligo was significantly lower than that in the control group. There was no significant difference in the serum level of TNF-α between vitiligo and healthy controls. Conclusion : An increase in the production of proinflammatory cytokines such as IL-6 and IL-2 in vitiligo patients may play an important role in melanocytic cytotoxicity. Thus, we speculate that the cytokine production of epidermal microenvironment may be involved in vitiligo.

  12. Silent IL2RG Gene Editing in Human Pluripotent Stem Cells.

    Science.gov (United States)

    Li, Li B; Ma, Chao; Awong, Geneve; Kennedy, Marion; Gornalusse, German; Keller, Gordon; Kaufman, Dan S; Russell, David W

    2016-03-01

    Many applications of pluripotent stem cells (PSCs) require efficient editing of silent chromosomal genes. Here, we show that a major limitation in isolating edited clones is silencing of the selectable marker cassette after homologous recombination and that this can be overcome by using a ubiquitous chromatin opening element (UCOE) promoter-driven transgene. We use this strategy to edit the silent IL2RG locus in human PSCs with a recombinant adeno-associated virus (rAAV)-targeting vector in the absence of potentially genotoxic, site-specific nucleases and show that IL2RG is required for natural killer and T-cell differentiation of human PSCs. Insertion of an active UCOE promoter into a silent locus altered the histone modification and cytosine methylation pattern of surrounding chromatin, but these changes resolved when the UCOE promoter was removed. This same approach could be used to correct IL2RG mutations in X-linked severe combined immunodeficiency patient-derived induced PSCs (iPSCs), to prevent graft versus host disease in regenerative medicine applications, or to edit other silent genes. PMID:26444081

  13. Cystatin F regulates proteinase activity in IL-2-activated natural killer cells.

    Science.gov (United States)

    Maher, Katarina; Konjar, Spela; Watts, Colin; Turk, Boris; Kopitar-Jerala, Natasa

    2014-01-01

    Cystatin F is a unique member of the cystatin family of cysteine protease inhibitors, which is synthesized as an inactive dimer and it is activated by N-terminal cleavage in the endolysosomes. It is expressed in the cells of the immune system: myeloid cells and the cells involved in target cell killing: natural killer (NK) cells and cytotoxic T cells (CTLs). Upon activation of the NK cells with interleukin 2 (IL-2), cystatin F was found upregulated and co-localized in cytotoxic granules with cathepsin C (CatC) and CatV. However, cystatin F inhibits the CatC in cells only when its N-terminal part is processed. Although cystatin F could inhibit both CatV and CatC, the IL-2 stimulation of the YT cells resulted in an increased CatV activity, while the CatC activity was unchanged. The incubation of IL-2 activated NK cells with a cysteine proteinase inhibitor E-64d increased the cystatin F dimer formation. Our results suggest that cystatin F not only inhibits CatV, but it is processed by the CatV in order to inhibit the CatC activity in cytotoxic granules. The regulation of the CatC activity in the cytotoxic granules of the NK cells by the cystatin F could be important for the processing and activation of granule-associated serine proteases - granzymes.

  14. Regulation of IL-2 induced proliferation and cytotoxicity in human natural killer cells by monoclonal antibodies

    International Nuclear Information System (INIS)

    Natural killer (NK) activity is mediated by a subpopulation of cells termed large granular lymphocytes (LGL), which exhibit cytotoxic activity against a variety of tumor targets. LGL express OKT8, OKT9, OKT10, OKT11, 3G8 (FcγR), OKM1, NKH1. The addition of recombinant IL-2 (rIL-2), increases cytotoxicity, induces IFN-γ production and leads to LGL proliferation. Since monoclonal antibodies (MoAb) represent highly specific probes to analyze possible surface molecules, they have studied the role of various MoAbs in the regulation of cytotoxicity, proliferation, and secretory function of purified LGL. LGL were isolated from nonadherent human peripheral blood leukocytes on discontinuous Percoll density gradients, followed by 290C E-rosette depletion of contaminating T cells. These preparations were ≥ 85% LGL and contained ≥ 5% OKT3+ cells. Using a limiting dilution assay, purified LGL were incubated with rIL-2 and the MoAbs (10 μg/ml) for 7 days. These cells were tested for cytotoxicity against K562 in a 51Cr- release assay, and for proliferation as determined by 3H-thymidine incorporation. Results indicate that the OKT9 antibody inhibited both the cytotoxicity and proliferation. MoAb against LGl markers (OKT11, OKT8, OKM1, 3G8, and NKH1) had no effect on cytotoxicity or proliferation. Unlike the T cell receptor complex (with OKT3), the surface molecules examined do not regulate LGL lysis or proliferation

  15. IL-2, IL-4, IFN-γ or TNF-α enhances BAFF-stimulated cell viability and survival by activating Erk1/2 and S6K1 pathways in neoplastic B-lymphoid cells.

    Science.gov (United States)

    Gui, Lin; Zeng, Qingyu; Xu, Zhigang; Zhang, Hai; Qin, Shanshan; Liu, Chunxiao; Xu, Chong; Qian, Zhou; Zhang, Shuangquan; Huang, Shile; Chen, Long

    2016-08-01

    B-cell activating factor of the TNF family (BAFF) has been documented to act as a critical factor in the development of aggressive B lymphocytes and autoimmune diseases. However, the effect of various cytokines on BAFF-elicited neoplastic B-lymphoid cells is not known. In this study, we exhibited that administration of human soluble BAFF (hsBAFF), IL-2, IL-4, IFN-γ, or TNF-α alone increased cell viability and survival in Raji cells concentration-dependently, yet a more robust viability/survival was seen in the cells co-treatment of IL-2, IL-4, IFN-γ, or TNF-α with hsBAFF, respectively. Further research revealed that both Erk1/2 and S6K1 signaling pathways were essential for IL-2, IL-4, IFN-γ, or TNF-α enhancement of the viability/survival in the hsBAFF-stimulated cells, as inhibition of Erk1/2 with U0126 or down-regulation of Erk1/2, or blockage of S6K1 with rapamycin or silencing S6K1, or silencing S6K1/Erk1/2, respectively, reduced the cell viability/survival in the cells treated with/without hsBAFF±IL-2, IL-4, IFN-γ, or TNF-α. These findings indicate that IL-2, IL-4, IFN-γ or TNF-α enhances BAFF-stimulated cell viability/survival by activating Erk1/2 and S6K1 signaling in neoplastic B-lymphoid cells. Our data suggest that modulation of IL-2, IL-4, IFN-γ and/or TNF-α levels, or inhibitors of Erk1/2 or S6K1 may be a new approach to prevent BAFF-induced aggressive B-cell malignancies.

  16. Estabishment of A Human Liver Cancer Cell Line Transfected with IL-2 cDNA and Its Biologic Activity

    Institute of Scientific and Technical Information of China (English)

    孙跃明; 王学浩; 杜竞辉

    2001-01-01

    Objective To obtain IL-2 gene transfected human liver cancer cells and study IL-2 expression and its biologic activity in vivo. Methods Human liver cancer cells SMMC-7721 were cocultured with recombinant retroviral vector LNC-IL-2,and screening was performed in G418 medium.The exogenous IL-2 cDNA at the DNA,RNA,and protein levels were determined by using dot hybridization,PR-PCR and MTT methods respectively.The tumorigenesis and antitumorigenesis of the screened liver cancer cell with subcutaneous injection in nude mice were observed. Results and Conclusion The IL-2 cDNA was successfully integrated into SMMC-7721 cell genomic DNA and continuously expressed for more than 88 days.Subcutaneous vaccination of the nude mice with transfected cells revealed an obvious suppression of its tumorigenicity,and could induce antitumor activity in vivo.

  17. Elevation in liver enzymes is associated with increased IL-2 and predicts severe outcomes in clinically apparent dengue virus infection.

    Science.gov (United States)

    Senaratne, Thamarasi; Carr, Jillian; Noordeen, Faseeha

    2016-07-01

    The objective of the present study was to assess the circulating TNF-α and IL-2 levels in dengue virus (DENV) infected patients and to correlate these with clinical severity of DENV infections. A single analyte quantitative immunoassay was used to detect TNF-α and IL-2 in 24 dengue fever (DF) and 43 dengue haemorrhagic fever (DHF) patients, 15 healthy adults and 6 typhoid patients. The mean TNF-α and IL-2 levels of DENV- infected patients were higher than that of healthy adults and typhoid patients. No significant difference in TNF-α levels was noted between DF and DHF patients (p=0.5) but a significant increase in IL-2 levels was observed in DHF compared with DF patients (mean of DF=59.7pg/mL, mean of DHF=166.9pg/mL; p=0.02). No significant association of TNF-α or IL-2 levels was noted with packed cell volume (>45), thrombocytopenia, leucopenia or the presence of viraemia. The liver function tests measuring AST (aspartate aminotransferase) (p=0.01) and ALT (alanine aminotransferase) (p=0.02) levels were significantly elevated in DENV-infected patients. AST:ALT was significantly elevated in DHF/DSS (dengue shock syndrome) compared with DF patients. A significant positive linear correlation was noted between AST and IL-2 (r=0.31; p=0.01) and ALT and IL-2 elevations (r=0.2; p=0.02). Thus, AST and ALT levels correlate with both disease severity and circulating IL-2 levels. We suggest a role for circulating IL-2 in liver dysfunction and propose that a combined assessment of AST/ALT in conjunction with IL-2 at the early stages of symptomatic DENV infection may be useful to predict the severe forms of dengue. PMID:27155816

  18. Association of the IL2RA/CD25 Gene With Juvenile Idiopathic Arthritis

    Science.gov (United States)

    Hinks, Anne; Ke, Xiayi; Barton, Anne; Eyre, Steve; Bowes, John; Worthington, Jane; Thompson, Susan D; Langefeld, Carl D; Glass, David N; Thomson, Wendy

    2009-01-01

    Objective IL2RA/CD25, the gene for interleukin-2 receptor α, is emerging as a general susceptibility gene for autoimmune diseases because of its role in the development and function of regulatory T cells and the association of single-nucleotide polymorphisms (SNPs) within this gene with type 1 diabetes mellitus (DM), Graves' disease, rheumatoid arthritis (RA), and multiple sclerosis (MS). The aim of this study was to determine whether SNPs within the IL2RA/CD25 gene are associated with juvenile idiopathic arthritis (JIA). Methods Three SNPs within the IL2RA/CD25 gene, that previously showed evidence of an association with either RA, MS, or type 1 DM, were selected for genotyping in UK JIA cases (n = 654) and controls (n = 3,849). Data for 1 SNP (rs2104286) were also available from North American JIA cases (n = 747) and controls (n = 1,161). Association analyses were performed using Plink software. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated. Results SNP rs2104286 within the IL2RA/CD25 gene was significantly associated with UK JIA cases (OR for the allele 0.76 [95% CI 0.66–0.88], P for trend = 0.0002). A second SNP (rs41295061) also showed modest evidence for association with JIA (OR 0.80 [95% CI 0.63–1.0], P = 0.05). Association with rs2104286 was convincingly replicated in the North American JIA cohort (OR 0.84 [95% CI 0.65–0.99], P for trend = 0.05). Meta-analysis of the 2 cohorts yielded highly significant evidence of association with JIA (OR 0.76 [95% CI 0.62–0.88], P = 4.9 × 10−5). Conclusion These results provide strong evidence that the IL2RA/CD25 gene represents a JIA susceptibility locus. Further investigation of the gene using both genetic and functional approaches is now required. PMID:19116909

  19. Expression of sIL-2R before and after Chemotherapy in Patients with Breast Cancer

    Institute of Scientific and Technical Information of China (English)

    FAN Yuan-ming; SUN Zhi-jun

    2008-01-01

    Objective:To investigate the difference of peripheral blood sIL-2R before and after chemotherapy in breast cancer patients,and evaluate the clinical value of the sIL-2R in breast cancer's diagnosis and therapy.Methods:The peripheral blood sIL-2R levels of the breast cancer patients with or without chemotherapy were detected by ELISA.The healthy persons were made as the control group.Results:The slL-2R levels of the breast cancer patients were higher than that of the control group(P<0.05);the slL-2R's levels in Ⅰ~Ⅱ stage breast cancer were lower than that in Ⅲ~Ⅳ stag e breast cancer (P<0.05);the sIL-2R levels of the patients before chemotherapy were higher than that of the patients undergone chemotherapy(P<0.05);The level of the patient with chemotherapy was still higher than that of the control group(P<0.05);the sIL-2R levels of the patients whose chemotherapies were noneffective were higher than that of the patients received effective chemotherapies(P<0.05).There was no significant difference between the group with ER(+)or PR(+)and the group with ER(-)or PR(-)(P>0.05).Conclusion:The breast cancer patients have the high slL-2R levels.There is a close relationship between the cancer incidence and the patients,immune situation.The level of slL-2R could be a clinical index which Can be used for evaluating the cancer degree,because the higher levels of slL-2R can indicate that the immune ability of patient is worse.There is a significant difference between the slL-2R levels of the patients before chemotherapy and that of the patients undergone chemotherapy.

  20. Long-term effects of intermittent IL-2 in HIV infection: extended follow-up of the INSIGHT STALWART Study.

    Directory of Open Access Journals (Sweden)

    Norman Markowitz

    Full Text Available BACKGROUND: The Study of Aldesleukin with and without Antiretroviral Therapy (STALWART was designed to evaluate whether intermittent IL-2 alone or with peri-cycle ART increased CD4+ cell counts (and so delayed initiation of ART in HIV infected individuals having ≥ 300 CD4+ cells/mm(3 compared to untreated controls. When the results of two large clinical trials, ESPRIT and SILCAAT, showed no clinical benefit from IL-2 therapy, IL-2 administration was halted in STALWART. Because IL-2 recipients in STALWART experienced a greater number of opportunistic disease (OD or death and adverse events (AEs, participants were asked to consent to an extended follow-up phase in order to assess persistence of IL-2 effects. METHODOLOGY: Participants in this study were followed for clinical events and AEs every 4 months for 24 months. Unadjusted Cox proportional hazards models were used to summarize death, death or first OD event, and first grade 3 or 4 AE. PRINCIPAL FINDINGS: A total of 267 persons were enrolled in STALWART (176 randomized to the IL-2 arms and 91 to the no therapy arm; 142 individuals in the IL-2 group and 80 controls agreed to enter the extended follow-up study. Initiation of continuous ART was delayed in the IL-2 groups, but once started, resulted in similar CD4+ cell and viral load responses compared to controls. The hazard ratios (95% CI for IL-2 versus control during the extension phase for death or OD, grade 3 or 4 AE, and grade 4 AE were 1.45 (0.38, 5.45, 0.43 (0.24, 1.63 and 0.20 (0.04, 1.03, respectively. The hazard ratios for the AE outcomes were significantly lower during the extension than during the main study. CONCLUSIONS: Adverse events associated with IL-2 cycling did not persist upon discontinuation of IL-2. The use of IL-2 did not impact the subsequent response to initiation of cART.

  1. Detecting IL-2 and IL-10 to predicate treatment outcome for Graves disease%IL-2与IL-10浓度变化对Graves病治疗效果的预测

    Institute of Scientific and Technical Information of China (English)

    李春北

    2010-01-01

    目的:探讨IL-2和IL-10变化对Graves病(Graves disease,GD)治疗转归的影响.方法:以2007年1月至2008年12月入我院治疗的GD患者72例,分别于治疗前及治疗后5月检测IL-2和IL-10及甲状腺素水平,评价IL-2和IL-10水平与治疗转归的关系.结果:72例GD患者甲亢未控制者21例,甲亢控制者35例,早发性甲减者16例.IL-2在甲亢未控制者水平最高,甲亢控制者次之,而早发性甲减者最低(P0.05).结论:IL-2/IL-10比值越高甲亢越难控制,比值越低越易产生甲减;IL-2/IL-10是较好的GD治疗效果预测指标.

  2. {sup 18}F-FDG PET, genotype-corrected ACE and sIL-2R in newly diagnosed sarcoidosis

    Energy Technology Data Exchange (ETDEWEB)

    Keijsers, Ruth G.; Verzijlbergen, Fred J. [St. Antonius Hospital Nieuwegein, Department of Nuclear Medicine, P.O. Box 2500, Nieuwegein (Netherlands); Oyen, Wim J. [Radboud University Nijmegen Medical Center, Department of Nuclear Medicine, Nijmegen (Netherlands); Bosch, Jules M. van den; Grutters, Jan C. [St. Antonius Hospital Nieuwegein, Department of Pulmonology, Nieuwegein (Netherlands); Ruven, Henk J. [St. Antonius Hospital Nieuwegein, Department of Clinical Chemistry, Nieuwegein (Netherlands); Velzen-Blad, Heleen van [St. Antonius Hospital Nieuwegein, Department of Medical Microbiology and Immunology, Nieuwegein (Netherlands)

    2009-07-15

    Angiotensin-converting enzyme (ACE) and soluble interleukin-2 receptor (sIL-2R) are serological markers, widely used for determining sarcoidosis activity. {sup 18}F-FDG PET has proven to be a sensitive technique in the imaging of sarcoidosis. The aim of this study was to determine sensitivity of {sup 18}F-FDG PET, genotype-corrected ACE and sIL-2R in active sarcoidosis as well as their correlation. This retrospective study included 36 newly diagnosed, symptomatic sarcoidosis patients. ACE and sIL-2R levels were simultaneously obtained within 4 weeks of {sup 18}F-FDG PET. ACE was corrected for genotype and expressed as Z-score. {sup 18}F-FDG PET was visually evaluated and scored as positive or negative. Maximum and average standardized uptake values (SUV{sub max} and SUV{sub avg}) were compared with ACE and sIL-2R. {sup 18}F-FDG PET was found positive in 34 of 36 patients (94%). Thirteen patients (36%) showed an increased ACE with the highest sensitivity found in patients with the I/I genotype (67%). Seventeen patients (47%) showed an increased sIL-2R. No correlation was found between SUV and ACE or sIL-2R. Increased ACE and sIL-2R correlated with a positive {sup 18}F-FDG PET in 12 patients (92%) and 16 patients (94%), respectively. {sup 18}F-FDG PET is a very sensitive technique to assess active sarcoidosis, in contrast with ACE and sIL-2R, suggesting a pivotal role for {sup 18}F-FDG PET in future sarcoidosis assessment. (orig.)

  3. Qualitative Immune Modulation by Interleukin-2 (IL-2) Adjuvant Therapy in Immunological Non Responder HIV-Infected Patients

    OpenAIRE

    Francesca Sabbatini; Alessandra Bandera; Giulio Ferrario; Daria Trabattoni; Giulia Marchetti; Fabio Franzetti; Mario Clerici; Andrea Gori

    2010-01-01

    BACKGROUND: Treatment of HIV-infected patients with interleukin-2 (IL-2) produces significant increases in CD4 T cell counts; however an associated qualitative improvement in cells function has yet to be conclusively demonstrated. By measuring mycobacterial killing activity, we evaluated IL-2-mediated functional immune enhancement ex vivo in immunological non-responders (INRs). METHODS AND FINDINGS: PBMC from 12 immunological non-responders (INRs) (CD4+

  4. Anticancer immunotherapy by CTLA-4 blockade: obligatory contribution of IL-2 receptors and negative prognostic impact of soluble CD25.

    Science.gov (United States)

    Hannani, Dalil; Vétizou, Marie; Enot, David; Rusakiewicz, Sylvie; Chaput, Nathalie; Klatzmann, David; Desbois, Melanie; Jacquelot, Nicolas; Vimond, Nadège; Chouaib, Salem; Mateus, Christine; Allison, James P; Ribas, Antoni; Wolchok, Jedd D; Yuan, Jianda; Wong, Philip; Postow, Michael; Mackiewicz, Andrzej; Mackiewicz, Jacek; Schadendorff, Dirk; Jaeger, Dirk; Zörnig, Inka; Hassel, Jessica; Korman, Alan J; Bahjat, Keith; Maio, Michele; Calabro, Luana; Teng, Michele Wl; Smyth, Mark J; Eggermont, Alexander; Robert, Caroline; Kroemer, Guido; Zitvogel, Laurence

    2015-02-01

    The cytotoxic T lymphocyte antigen-4 (CTLA-4)-blocking antibody ipilimumab induces immune-mediated long-term control of metastatic melanoma in a fraction of patients. Although ipilimumab undoubtedly exerts its therapeutic effects via immunostimulation, thus far clinically useful, immunologically relevant biomarkers that predict treatment efficiency have been elusive. Here, we show that neutralization of IL-2 or blocking the α and β subunits of the IL-2 receptor (CD25 and CD122, respectively) abolished the antitumor effects and the accompanying improvement of the ratio of intratumoral T effector versus regulatory cells (Tregs), which were otherwise induced by CTLA-4 blockade in preclinical mouse models. CTLA-4 blockade led to the reduction of a suppressive CD4(+) T cell subset expressing Lag3, ICOS, IL-10 and Egr2 with a concomitant rise in IL-2-producing effector cells that lost FoxP3 expression and accumulated in regressing tumors. While recombinant IL-2 improved the therapeutic efficacy of CTLA-4 blockade, the decoy IL-2 receptor α (IL-2Rα, sCD25) inhibited the anticancer effects of CTLA-4 blockade. In 262 metastatic melanoma patients receiving ipilimumab, baseline serum concentrations of sCD25 represented an independent indicator of overall survival, with high levels predicting resistance to therapy. Altogether, these results unravel a role for IL-2 and IL-2 receptors in the anticancer activity of CTLA-4 blockade. Importantly, our study provides the first immunologically relevant biomarker, namely elevated serum sCD25, that predicts resistance to CTLA-4 blockade in patients with melanoma. PMID:25582080

  5. EXPRESSION OF IL-2 AND SIL-2R AND ALTERATION OF CELL IMMUNITY IN PATIENTS WITH HYPERTENSIVE CEREBRAL HEMORRHAGE

    Institute of Scientific and Technical Information of China (English)

    Zhang Yuelin; Qiu Shudong; Shi Wei; Dang Xiaojun

    2006-01-01

    Objective To study the expression of interleukin-2 (IL-2), soluble interleukin-2 receptor (sIL-2R),determine the alteration of erythrocytic immunity and T cell subgroup in the blood of outer circulation in patients with hypertensive cerebral hemorrhage so and to probe into the relationship between them, and to explore the clinical significance. Methods Enzyme linked immnunosorbent assay (ELISA) was used to determine the content of IL-2 and sIL-2R. The immunoadsorption was employed to examine the erythrocytic immune activity and its regulating function.Streptavidin-peroxidase(S-P) was used to determine the cell number of CD3 (cluster of differentiation3), CD4 and CD8. Results The content of IL-2 in the group with hypertensive cerebral hemorrhage was significantly lower than that in the control group (P<0.01), and the content of sIL-2R increased. Red blood cell C3b receptor (RBC. C3b R)and RBC immune adherence enhancing factor (RFEB) dropped greatly (P<0.01), while RBC immune complex rosette (RBC. ICR) and RBC immune adherence inhibiting factor (RFIR) increased greatly. The cell number of CD3 and CD4decreased (P<0.01) and there was no obvious change in CD8 (P<0.05). Conclusion The decrease of immune function was observed in patients with hypertensive cerebral hemorrhage. The determination of the content of IL-2, sIL-2R, erythrocytic immunity and the activity of T subgroup has an important clinical significance in the occurrence,development, treatment, and prognosis of hypertensive cerebral hemorrhage.

  6. Safety and immunogenicity of Salmonella typhimurium expressing C-terminal truncated human IL-2 in a murine model

    Directory of Open Access Journals (Sweden)

    Brent Sorenson

    2010-03-01

    Full Text Available Brent Sorenson, Kaysie Banton, Lance Augustin, Sean Barnett, Karen McCulloch, Joshua Dorn, Natalie Frykman, Arnold Leonard, Daniel SaltzmanDepartment of Surgery, University of Minnesota Medical School, Minneapolis, MN, USAAbstract: Salmonella enterica serovar Typhimurium preferentially colonizes tumors in vivo and has proven to be an effective biologic vector. The attenuated S. enterica Typhimurium strain χ4550 was engineered to express truncated human interleukin-2 and renamed SalpIL2. Previously, we observed that a single oral administration of SalpIL2 reduced tumor number and volume, while significantly increasing local and systemic natural killer (NK cell populations in an experimental metastasis model. Here we report that in nontumor-bearing mice, a single oral dose of SalpIL2 resulted in increased splenic cytotoxic T and NK cell populations that returned to control levels by 4 weeks post oral administration. Though SalpIL2 was detected in mouse tissues for up to 10 weeks, no prolonged alterations in peripheral blood serum chemistry or complete blood cell counts were observed. Similarly, comparative histopathological analysis of tissues revealed no significant increase in pyogranulomas in SalpIL2-treated animals with respect to saline controls. In Rag-1 knockout mice, which have severely impaired B and T cell function, SalpIL2 reduced growth of hepatic metastases. Furthermore, SalpIL2 altered expression of several proinflammatory cytokines and chemokines in the serum of mice with pulmonary osteosarcoma metastases. These data further suggest that SalpIL2 is avirulent and induces a cell-mediated antitumor response.Keywords: Salmonella Typhimurium, natural killer cells, interleukin-2

  7. Stromal cell-derived factors in Duchenne muscular dystrophy

    OpenAIRE

    Abdel-Salam, E.; Ehsan Abdel-Meguid, I.; Shatla, R.; Korraa, S. S.

    2010-01-01

    Duchenne muscular dystrophy (DMD) is characterized by increased muscle damage and an abnormal blood flow after muscle contraction leading to a state of functional ischemia. Abundant evidence suggests that endothelial circulating progenitor cells (EPCs) play an important role in mediating vascular and muscle repair mechanisms and that the stromal cell-derived factor (SDF)-1 α chemokine is responsible for both progenitor cell mobilization from the bone marrow to peripheral blood and homing to t...

  8. Natural Helper cells derive from lymphoid progenitors1

    OpenAIRE

    Yang, Qi; Saenz, Steven A.; Zlotoff, Daniel A.; Artis, David; Bhandoola, Avinash

    2011-01-01

    Natural Helper (NH) cells are recently discovered innate immune cells that confer protective type 2 immunity during helminth infection and mediate influenza induced airway hypersensitivity. Little is known about the ontogeny of NH cells. We now report NH cells derive from bone marrow lymphoid progenitors. Using RAG-1Cre/ROSA26YFP mice, we show that the majority of NH cells are marked with a history of RAG-1 expression, implying lymphoid developmental origin. The development of NH cells depend...

  9. PGC-1α-Dependent Mitochondrial Adaptation Is Necessary to Sustain IL-2-Induced Activities in Human NK Cells.

    Science.gov (United States)

    Miranda, Dante; Jara, Claudia; Ibañez, Jorge; Ahumada, Viviana; Acuña-Castillo, Claudio; Martin, Adrian; Córdova, Alexandra; Montoya, Margarita

    2016-01-01

    Human Natural Killer (NK) cells are a specialized heterogeneous subpopulation of lymphocytes involved in antitumor defense reactions. NK cell effector functions are critically dependent on cytokines and metabolic activity. Among various cytokines modulating NK cell function, interleukin-2 (IL-2) can induce a more potent cytotoxic activity defined as lymphokine activated killer activity (LAK). Our aim was to determine if IL-2 induces changes at the mitochondrial level in NK cells to support the bioenergetic demand for performing this enhanced cytotoxic activity more efficiently. Purified human NK cells were cultured with high IL-2 concentrations to develop LAK activity, which was assessed by the ability of NK cells to lyse NK-resistant Daudi cells. Here we show that, after 72 h of culture of purified human NK cells with enough IL-2 to induce LAK activity, both the mitochondrial mass and the mitochondrial membrane potential increased in a PGC-1α-dependent manner. In addition, oligomycin, an inhibitor of ATP synthase, inhibited IL-2-induced LAK activity at 48 and 72 h of culture. Moreover, the secretion of IFN-γ from NK cells with LAK activity was also partially dependent on PGC-1α expression. These results indicate that PGC-1α plays a crucial role in regulating mitochondrial function involved in the maintenance of LAK activity in human NK cells stimulated with IL-2. PMID:27413259

  10. A BCR/ABL-hIL-2 DNA Vaccine Enhances the Immune Responses in BALB/c Mice

    Directory of Open Access Journals (Sweden)

    Yanan Qin

    2013-01-01

    Full Text Available The use of a DNA vaccine encoding the BCR/ABL fusion gene is thought to be a promising approach for patients with chronic myeloid leukemia (CML to eradicate minimal residual disease after treatment with chemotherapy or targeted therapy. In this study, our strategy employs genetic technology to create a DNA vaccine encoding the BCR/ABL fusion and human interleukin-2 (hIL-2 genes. The successfully constructed plasmids BCR/ABL-pIRES-hIL-2, BCR/ABL-pIRES, and pIRES-hIL-2 were delivered intramuscularly to BALB/c mice at 14-day intervals for three cycles. The transcription and expression of the BCR/ABL and hIL-2 genes were found in the injected muscle tissues. The interferon-γ (IFN-γ serum levels were increased, and the splenic CD4+/CD8+ T cell ratio was significantly decreased in the BCR/ABL-pIRES-hIL-2-injected mice. Furthermore, specific antibodies against K562 cells could be detected by indirect immunofluorescence. These results indicate that a DNA vaccine containing BCR/ABL and hIL-2 together may elicit increased in vivo humoral and cellular immune responses in BALB/c mice.

  11. Human retinal pigment epithelial cells inhibit proliferation and IL2R expression of activated T cells

    DEFF Research Database (Denmark)

    Kaestel, Charlotte G; Jørgensen, Annette; Nielsen, Mette;

    2002-01-01

    The purpose of this study was to characterize the effects of human retinal pigment epithelial (RPE) cells on activated T cells. Activated T cells were cocultured with adult and foetal human RPE cells whereafter apoptosis and proliferation were determined by flow cytometry and (3)H......-Thymidine incorporation assay, respectively. T cells and RPE cells were cultured directly together or in a transwell system for determination of the effect of cell contact. The importance of cell surface molecules was examined by application of a panel of blocking antibodies (CD2, CD18, CD40, CD40L, CD54, CD58......) in addition to use of TCR negative T cell lines. The expression of IL2R-alpha -beta and -gamma chains of activated T cells was analysed by flow cytometry after incubation of T cells alone or with RPE cells. Human RPE cells were found to inhibit the proliferation of activated T cells by a cell contact...

  12. Pathogen sensing pathways in human embryonic stem cell derived-endothelial cells: role of NOD1 receptors.

    Directory of Open Access Journals (Sweden)

    Daniel M Reed

    Full Text Available Human embryonic stem cell-derived endothelial cells (hESC-EC, as well as other stem cell derived endothelial cells, have a range of applications in cardiovascular research and disease treatment. Endothelial cells sense Gram-negative bacteria via the pattern recognition receptors (PRR Toll-like receptor (TLR-4 and nucleotide-binding oligomerisation domain-containing protein (NOD-1. These pathways are important in terms of sensing infection, but TLR4 is also associated with vascular inflammation and atherosclerosis. Here, we have compared TLR4 and NOD1 responses in hESC-EC with those of endothelial cells derived from other stem cells and with human umbilical vein endothelial cells (HUVEC. HUVEC, endothelial cells derived from blood progenitors (blood outgrowth endothelial cells; BOEC, and from induced pluripotent stem cells all displayed both a TLR4 and NOD1 response. However, hESC-EC had no TLR4 function, but did have functional NOD1 receptors. In vivo conditioning in nude rats did not confer TLR4 expression in hESC-EC. Despite having no TLR4 function, hESC-EC sensed Gram-negative bacteria, a response that was found to be mediated by NOD1 and the associated RIP2 signalling pathways. Thus, hESC-EC are TLR4 deficient but respond to bacteria via NOD1. This data suggests that hESC-EC may be protected from unwanted TLR4-mediated vascular inflammation, thus offering a potential therapeutic advantage.

  13. Phase I clinical trial combining imatinib mesylate and IL-2: HLA-DR(+) NK cell levels correlate with disease outcome.

    Science.gov (United States)

    Chaput, Nathalie; Flament, Caroline; Locher, Clara; Desbois, Mélanie; Rey, Annie; Rusakiewicz, Sylvie; Poirier-Colame, Vichnou; Pautier, Patricia; Le Cesne, Axel; Soria, Jean-Charles; Paci, Angelo; Rosenzwajg, Michelle; Klatzmann, David; Eggermont, Alexander; Robert, Caroline; Zitvogel, Laurence

    2013-02-01

    We performed a Phase I clinical trial from October 2007 to October 2009, enrolling patients affected by refractory solid tumors, to determine the maximum tolerated dose (MTD) of interleukin (IL)-2 combined with low dose cyclophosphamide (CTX) and imatinib mesylate (IM). In a companion paper published in this issue of OncoImmunology, we show that the MTD of IL-2 is 6 MIU/day for 5 consecutive days, and that IL-2 increases the impregnation of both IM and of its main metabolite, CGP74588. Among the secondary objectives, we wanted to determine immunological markers that might be associated with progression-free survival (PFS) and/or overall survival (OS). The combination therapy markedly reduced the absolute counts of B, CD4(+) T and CD8(+) T cells in a manner that was proportional to IL-2 dose. There was a slight (less than 2-fold) increase in the proportion of regulatory T cells (Tregs) among CD4(+) T cells in response to IM plus IL-2. The natural killer (NK)-cell compartment was activated, exhibiting a significant upregulation of HLA-DR, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and CD56. The abundance of HLA-DR(+) NK cells after one course of combination therapy positively correlated with both PFS and OS. The IL-2-induced rise of the CD4(+):CD8(+) T-cell ratio calculated after the first cycle of treatment was also positively associated with OS. Overall, the combination of IM and IL-2 promoted the rapid expansion of HLA-DR(+) NK cells and increased the CD4(+):CD8(+) T-cell ratio, both being associated with clinical benefits. This combinatorial regimen warrants further investigation in Phase II clinical trials, possibly in patients affected by gastrointestinal stromal tumors, a setting in which T and NK cells may play an important therapeutic role. PMID:23525357

  14. Regulatory T cells negatively affect IL-2 production of effector T cells through CD39/adenosine pathway in HIV infection.

    Directory of Open Access Journals (Sweden)

    Mohammad-Ali Jenabian

    Full Text Available The mechanisms by which Regulatory T cells suppress IL-2 production of effector CD4+ T cells in pathological conditions are unclear. A subpopulation of human Treg expresses the ectoenzyme CD39, which in association with CD73 converts ATP/ADP/AMP to adenosine. We show here that Treg/CD39+ suppress IL-2 expression of activated CD4+ T-cells more efficiently than Treg/CD39-. This inhibition is due to the demethylation of an essential CpG site of the il-2 gene promoter, which was reversed by an anti-CD39 mAb. By recapitulating the events downstream CD39/adenosine receptor (A2AR axis, we show that A2AR agonist and soluble cAMP inhibit CpG site demethylation of the il-2 gene promoter. A high frequency of Treg/CD39+ is associated with a low clinical outcome in HIV infection. We show here that CD4+ T-cells from HIV-1 infected individuals express high levels of A2AR and intracellular cAMP. Following in vitro stimulation, these cells exhibit a lower degree of demethylation of il-2 gene promoter associated with a lower expression of IL-2, compared to healthy individuals. These results extend previous data on the role of Treg in HIV infection by filling the gap between expansion of Treg/CD39+ in HIV infection and the suppression of CD4+ T-cell function through inhibition of IL-2 production.

  15. Different interleukin 2 receptor beta-chain tyrosines couple to at least two signaling pathways and synergistically mediate interleukin 2-induced proliferation.

    OpenAIRE

    Friedmann, M C; Migone, T S; Russell, S M; Leonard, W J

    1996-01-01

    One of the earliest events induced by interleukin 2 (IL-2) is tyrosine phosphorylation of cellular proteins, including the IL-2 receptor beta chain (IL-2Rbeta). Simultaneous mutation of three tyrosines (Y338, Y392, and Y510) in the IL-2Rbeta cytoplasmic domain abrogated IL-2-induced proliferation, whereas mutation of only Y338 or of Y392 and Y510 inhibited proliferation only partially. While Y392 and Y510 were critical for IL-2-induced activation of signal transducers and activators of transc...

  16. Neural stem cell-derived exosomes mediate viral entry

    Directory of Open Access Journals (Sweden)

    Sims B

    2014-10-01

    Full Text Available Brian Sims,1,2,* Linlin Gu,3,* Alexandre Krendelchtchikov,3 Qiana L Matthews3,4 1Division of Neonatology, Department of Pediatrics, 2Department of Cell, Developmental, and Integrative Biology, 3Division of Infectious Diseases, Department of Medicine, 4Center for AIDS Research, University of Alabama at Birmingham, Birmingham, AL, USA *These authors contributed equally to this work Background: Viruses enter host cells through interactions of viral ligands with cellular receptors. Viruses can also enter cells in a receptor-independent fashion. Mechanisms regarding the receptor-independent viral entry into cells have not been fully elucidated. Exosomal trafficking between cells may offer a mechanism by which viruses can enter cells.Methods: To investigate the role of exosomes on cellular viral entry, we employed neural stem cell-derived exosomes and adenovirus type 5 (Ad5 for the proof-of-principle study. Results: Exosomes significantly enhanced Ad5 entry in Coxsackie virus and adenovirus receptor (CAR-deficient cells, in which Ad5 only had very limited entry. The exosomes were shown to contain T-cell immunoglobulin mucin protein 4 (TIM-4, which binds phosphatidylserine. Treatment with anti-TIM-4 antibody significantly blocked the exosome-mediated Ad5 entry.Conclusion: Neural stem cell-derived exosomes mediated significant cellular entry of Ad5 in a receptor-independent fashion. This mediation may be hampered by an antibody specifically targeting TIM-4 on exosomes. This set of results will benefit further elucidation of virus/exosome pathways, which would contribute to reducing natural viral infection by developing therapeutic agents or vaccines. Keywords: neural stem cell-derived exosomes, adenovirus type 5, TIM-4, viral entry, phospholipids

  17. Construction of recombinant attenuated Salmonella typhimurium DNA vaccine expressing H pylori ureB and IL-2

    Institute of Scientific and Technical Information of China (English)

    Can Xu; Zhao-Shen Li; Yi-Qi Du; Yan-Fang Gong; Hua Yang; Bo Sun; Jing Jin

    2007-01-01

    AIM: To construct a recombinant live attenuated Salmonella typhimurium DNA vaccine encoding H pylori ureB gene and mouse IL-2 gene and to detect its immunogenicity in vitro and in vivo.METHODS: H pylori ureB and mouse IL-2 gene fragments were amplified by potymerase chain reaction (PCR) and cloned into pUCmT vector. DNA sequence of the amplified ureB and IL-2 genes was assayed, then cloned into the eukaryotic expression vector pIRES through enzyme digestion and ligation reactions resulting in pIRES-ureB and pIRES-ureB-IL-2. The recombinant plasmids were used to transform competent E. Coli DH5a, and the positive clones were screened by PCR and restriction enzyme digestion. Then, the recombinant pIRES-ureB and pIRES-ureB-IL-2 were used to transform LB5000 and the recombinant plasmids extracted from LB5000 were finally introduced into the final host SL7207. After that, recombinant strains were grown in vitro repeatedly. In order to detect the immunogenicity of the vaccine in vitro, pIRES-ureB and pIRES-ureB-IL-2 were transfected to COS-7 cells using Lipofectamine TM 2000, the immunogenicity of expressed UreB and IL-2 proteins was assayed with SDS-PAGE and Western blot. C57BL/6 mice were orally immunized with 1 x 108 recombinant attenuated Salmonella typhimurium DNA vaccine. Four weeks after vaccination, mice were challenged with 1 x 107 CFU of live Hpylori SSI. Mice were sacrificed and the stomach was isolated for examination of H pylori 4 wk post-challenge.RESULTS: The 1700 base pair ureB gene fragment amplified from the genomic DNA was consistent with the sequence of H pylori ureB by sequence analysis. The amplified 510 base pair fragment was consistent with the sequence of mouse IL-2 in gene bank. It was confirmed by PCR and restriction enzyme digestion that H pylori ureB and mouse IL-2 genes were inserted into the eukaryotic expression vector pIRES. The experiments in vitro snowed that stable recombinant live attenuated Salmonella typhimurium DNA vaccine carrying

  18. The high-dose aldesleukin (IL-2) "select" trial: a trial designed to prospectively validate predictive models of response to high-dose IL-2 treatment in patients with metastatic renal cell carcinoma.

    Science.gov (United States)

    Clement, Jessica M; McDermott, David F

    2009-08-01

    For patients with metastatic renal cell carcinoma (RCC), the prognosis is poor. Despite the recent approval of drugs such as sorafenib, sunitinib, and temsirolimus, durable remissions of metastatic disease are rare. This is largely due to the fact that these drugs, while effective, do not result in the eradication of disease. In 1992, the FDA approved the use of high-dose interleukin-2 (IL-2) for the treatment of patients with metastatic RCC because of a small number of patients that achieved durable responses. However, IL-2 has not become a mainstay of treatment because of the expense and toxicity associated with this therapy. This review article discusses a phase II trial that investigates predictive biomarkers that might help clinicians identify the patient population with metastatic RCC that would benefit from IL-2 therapy and therefore limit patients who receive this toxic therapy to those most likely to benefit. PMID:19692326

  19. Stable Expression of Hantavirus H8205 Strain G1/IL-2 Gene and Immune Protection of the Fusion Gene

    Institute of Scientific and Technical Information of China (English)

    XIONG Ying; YUAN Yuan; JIA Min; YU Bing; HUANG Hanju

    2007-01-01

    To explore the feasibility of stable expression of Hantavirus H8205 strain G1 segment and human IL-2 fusion gene in Vero cells, and to examine the immune protection effects on mice vaccinated with this recombinant eukaryotic expression vector containing Hantavirus G1 gene and IL-2 gene. With the help of lipofectamine, the Vero cells were transfected with pcDNA3.1/HisB-IL-2-G1 and the positive cells were selected by G418. IFAT and SDS-PAGE electrophoresis were used to determine the stable transfection and expression of recombinant protein.Each mouse was inoculated with plasmids intramuscularly (i.m.) three times, 2 boosts were given at 2-week intervals, serum anti-hantavirus antibodies were detected by ELISA and neutralizing antibodies (NAb) were detected by Plaque Reduction Neutralization Test. The fusion protein expressed in Vero cells was 78 kD, corresponding to the estimated molecular size. The neutralizing antibody titers of mice with pcDNA3.1/HisB-IL-2-G1 were 1:20-1:80. IL-2/G1 fusion gene could be transferred in Vero cells and stably express the fusion protein. Specific humeral immune responses in mice can be induced with the recombinant eukaryotic expression vector containing the fusion gene, which lays the foundation for further development of therapeutic HTNV vaccine.

  20. The biologic activity of melittin-88ArgIL-2 chimeric protein in vitro%蜂毒素与基因变构IL-2嵌合蛋白的体外生物活性研究

    Institute of Scientific and Technical Information of China (English)

    宋旭霞; 刘明军; 王斌; 钱冬萌; 闫志勇; 丁守怡

    2009-01-01

    目的:IL-2具有多种与增强免疫功能相关的生物学活性,蜂毒素对多种肿瘤细胞有杀伤作用.文中研究蜂毒素与基因变构IL-2(melittin-88ArgIL-2)嵌合蛋白在体外的生物学活性. 方法:用甲基噻唑基四唑(MTT)法研究纯化制备的melittin-88ArgIL-2嵌合蛋白在体外对T细胞增殖和NK细胞杀伤活性的影响及对人卵巢癌细胞SKOV3生长抑制的作用. 结果:melittin-88ArgIL-2嵌合蛋白在体外可促进T细胞的增值,增强NK细胞的杀伤活性,抑制人卵巢癌细胞SKOV3的生长增殖. 结论:melittin-88ArgIL-2嵌合蛋白在体外具有一定的增强免疫功能和抗肿瘤活性.值得进一步对其体内的生物学活性进行研究,为大规模制备的中试放大研究和临床前期研究提供了资料.

  1. Celecoxib offsets the negative renal influences of cyclosporine via modulation of the TGF-β1/IL-2/COX-2/endothelin ET{sub B} receptor cascade

    Energy Technology Data Exchange (ETDEWEB)

    El-Gowelli, Hanan M. [Pharmacology and Toxicology, Faculty of Pharmacy, Alexandria University, Alexandria (Egypt); Helmy, Maged W.; Ali, Rabab M. [Pharmacology and Toxicology, Faculty of Pharmacy, Pharos University, Alexandria (Egypt); El-Mas, Mahmoud M., E-mail: mahelm@hotmail.com [Pharmacology and Toxicology, Faculty of Pharmacy, Alexandria University, Alexandria (Egypt)

    2014-03-01

    Endothelin (ET) signaling provokes nephrotoxicity induced by the immunosuppressant drug cyclosporine A (CSA). We tested the hypotheses that (i): celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, counterbalances renal derangements caused by CSA in rats and (ii) the COX-2/endothelin ET{sub B} receptor signaling mediates the CSA-celecoxib interaction. Ten-day treatment with CSA (20 mg/kg/day) significantly increased biochemical indices of renal function (serum urea, creatinine), inflammation (interleukin-2, IL-2) and fibrosis (transforming growth factor-β{sub 1}, TGF-β{sub 1}). Histologically, CSA caused renal tubular atrophy along with interstitial fibrosis. These detrimental renal effects of CSA were largely reduced in rats treated concurrently with celecoxib (10 mg/kg/day). We also report that cortical glomerular and medullary tubular protein expressions of COX-2 and ET{sub B} receptors were reduced by CSA and restored to near-control values in rats treated simultaneously with celecoxib. The importance of ET{sub B} receptors in renal control and in the CSA-celecoxib interaction was further verified by the findings (i) most of the adverse biochemical, inflammatory, and histopathological profiles of CSA were replicated in rats treated with the endothelin ET{sub B} receptor antagonist BQ788 (0.1 mg/kg/day, 10 days), and (ii) the BQ788 effects, like those of CSA, were alleviated in rats treated concurrently with celecoxib. Together, the data suggest that the facilitation of the interplay between the TGF-β1/IL-2/COX-2 pathway and the endothelin ET{sub B} receptors constitutes the cellular mechanism by which celecoxib ameliorates the nephrotoxic manifestations of CSA in rats. - Highlights: • Celecoxib abolishes nephrotoxic manifestations of CSA in rats. • Blockade of ETB receptors by BQ788 mimicked the nephrotoxic effects of CSA. • CSA or BQ788 reduces renal protein expression of COX-2 and endothelin ETB receptors. • Enhanced TGFβ1/IL-2/COX2/ETB

  2. Functional Properties of Human Stem Cell-Derived Neurons in Health and Disease

    Directory of Open Access Journals (Sweden)

    Jason P. Weick

    2016-01-01

    Full Text Available Stem cell-derived neurons from various source materials present unique model systems to examine the fundamental properties of central nervous system (CNS development as well as the molecular underpinnings of disease phenotypes. In order to more accurately assess potential therapies for neurological disorders, multiple strategies have been employed in recent years to produce neuronal populations that accurately represent in vivo regional and transmitter phenotypes. These include new technologies such as direct conversion of somatic cell types into neurons and glia which may accelerate maturation and retain genetic hallmarks of aging. In addition, novel forms of genetic manipulations have brought human stem cells nearly on par with those of rodent with respect to gene targeting. For neurons of the CNS, the ultimate phenotypic characterization lies with their ability to recapitulate functional properties such as passive and active membrane characteristics, synaptic activity, and plasticity. These features critically depend on the coordinated expression and localization of hundreds of ion channels and receptors, as well as scaffolding and signaling molecules. In this review I will highlight the current state of knowledge regarding functional properties of human stem cell-derived neurons, with a primary focus on pluripotent stem cells. While significant advances have been made, critical hurdles must be overcome in order for this technology to support progression toward clinical applications.

  3. Requirement for noncognate interaction with T cells for the activation of B cell immunoglobulin secretion by IL-2

    DEFF Research Database (Denmark)

    Owens, T

    1991-01-01

    23.1+ TH1 clone E9.D4 in F23.1 (anti-T cell receptor V-beta 8)-coated microwells. This induced polyclonal B cell activation to enter cell cycle (thymidine incorporation) at 2 days and to secrete immunoglobulin at 5 days. An anti-IL-2 mAb (S4B6) inhibited antibody production completely. Anti-IL-2 did...... not inhibit either LPS-induced B cell responses, or T cell activation (measured as IL-3 secretion). Anti-IL-2 receptor (anti-Tac) mAbs also inhibited T-dependent B cell responses, without affecting LPS responses. An anti-IFN-gamma mAb partially inhibited Ig secretion, without affecting entry into...

  4. Matrix metalloproteinase-3 in odontoblastic cells derived from ips cells: unique proliferation response as odontoblastic cells derived from ES cells.

    Directory of Open Access Journals (Sweden)

    Taiki Hiyama

    Full Text Available We previously reported that matrix metalloproteinase (MMP-3 accelerates wound healing following dental pulp injury. In addition, we reported that a proinflammatory cytokine mixture (tumor necrosis factor-α, interleukin (IL-1β and interferon-γ induced MMP-3 activity in odontoblast-like cells derived from mouse embryonic stem (ES cells, suggesting that MMP-3 plays a potential unique physiological role in wound healing and regeneration of dental pulp in odontoblast-like cells. In this study, we tested the hypothesis that upregulation of MMP-3 activity by IL-1β promotes proliferation and apoptosis of purified odontoblast-like cells derived from induced pluripotent stem (iPS and ES cells. Each odontoblast-like cell was isolated and incubated with different concentrations of IL-1β. MMP-3 mRNA and protein expression were assessed using RT-PCR and western blotting, respectively. MMP-3 activity was measured using immunoprecipitation and a fluorescence substrate. Cell proliferation and apoptosis were determined using ELISA for BrdU and DNA fragmentation, respectively. siRNA was used to reduce MMP-3 transcripts in these cells. Treatment with IL-1β increased MMP-3 mRNA and protein levels, and MMP-3 activity in odontoblast-like cells. Cell proliferation was found to markedly increase with no changes in apoptosis. Endogenous tissue inhibitor of metalloproteinase (TIMP-1 and TIMP-2 were constitutively expressed during all experiments. The exocytosis inhibitor, Exo1, potently suppressed the appearance of MMP-3 in the conditioned medium. Treatment with siRNA against MMP-3 suppressed an IL-1β-induced increase in MMP-3 expression and activity, and also suppressed cell proliferation, but unexpectedly increased apoptosis in these cells (P<0.05. Exogenous MMP-3 was found to induce cell proliferation in odontoblast-like cells derived from iPS cells and ES cells. This siRNA-mediated increase in apoptosis could be reversed with exogenous MMP-3 stimulation (P<0

  5. Effect of catecholamines on IL-2 production and NK cytotoxicity of rats in vitro

    Institute of Scientific and Technical Information of China (English)

    Yu-ping PENG; Yi-hua QIU; Jian-lan JIANG; Jian-jun WANGTM

    2004-01-01

    AIM: To explore effects of exogenous and endogenous catecholamines on function of lymphocytes and primary mechanisms mediating the effects. METHODS: Splenocytes of rats were exposed to norepinephrine (NE), α- or β-adrenoceptor antagonists plus NE, or α-methyl-p-tyrosine (α-MT), and then concanavalin A (Con A)-induced intefleukin-2 (IL-2) production and natural killer (NK) cell cytotoxicity were determined by MTT assay and LDH assay, respectively. RESULTS: Optical density (OD) values of NE-treated groups, which reflected IL-2 production,were 0.63, 0.61, and 0.60, respectively for 1×10-10, 1×10-9, and 1×10-8 mol/L NE. They were all significantly reduced in comparison with control value of 0.68 (P<0.01). The effect of NE was blocked by either phentolamine (an α-adrenoceptor antagonist) or propanolol (a β-adrenoceptor antagonist). OD values of α-MT, an inhibitor of tyrosine hydroxylase, at doses of l× 10-10, 1× 10-9, and 1× 10-8 mol/L respectively were 0.71, 0.71, and 0.69, which were all notably higher than that of control (0.65, P<0.01). NK cytotoxicity was markedly attenuated by both NE and α-MT at the three doses mentioned above (17.69 %, 17.06 %, and 16.89 % versus 25.18 % for NE; 18.85 %,18.44 %, and 17.04 % versus 23.22 % for α-MT; all P<0.01). The suppression of NK cytotoxicity by NE was prevented by propranolol but not by phentolamine. CONCLUSION: Exogenous NE exerts a suppressive action in modulating functions of T and NK cells, with the former via both α- and β-adrenoceptor mediated mechanisms and the later mainly through β-adrenoceptors. Endogenous catecholamines synthesized by lymphocytes have also an autoregulatory effect on the lymphocytes themselves.

  6. In vitro assessment of choline dihydrogen phosphate (CDHP) as a vehicle for recombinant human interleukin-2 (rhIL-2)

    OpenAIRE

    Foureau, David M.; Vrikkis, Regina M.; Jones, Chase P.; Weaver, Katherine D.; Douglas R MacFarlane; Salo, Jonathan C.; McKillop, Iain H; Elliott, Gloria D.

    2012-01-01

    Choline dihydrogen phosphate (CDHP) is an ionic liquid reported to increase thermal stability of model proteins. The current work investigated CDHP effect on structural integrity and biological activity of recombinant human interleukin-2 (rhIL-2), a therapeutic protein used for treating advanced melanoma. In vitro CDHP biocompatibility was also evaluated using primary cell cultures, or B16-F10 cell line, chronically exposed to the ionic liquid. Formulation of rhIL-2 in an aqueous 680mM CDHP p...

  7. Safety and efficacy of subcutaneous and continuous intravenous infusion rIL-2 in patients with metastatic renal cell carcinoma

    DEFF Research Database (Denmark)

    Geertsen, P F; Gore, M E; Negrier, S;

    2004-01-01

    A retrospective analysis was conducted on data from four open-label, nonrandomised, phase II trials of recombinant interleukin-2 (rIL-2) in patients with metastatic renal cell carcinoma to compare the safety and efficacy of administration by subcutaneous (s.c.) and continuous intravenous (c...... required dose reductions because of toxicity (20 vs 82%). At the doses and within the schedules tested, this comparative analysis did not detect any difference in efficacy between s.c. and c.i.v. administration of rIL-2 in terms of overall survival, duration of response and response rate in patients with...

  8. Melittin-IL-2融合蛋白的抗肿瘤活性

    Institute of Scientific and Technical Information of China (English)

    刘明军; 宋旭霞; 王斌

    2009-01-01

    蜂毒素(Melittin)是欧洲蜜蜂(Apis mellifera)蜂毒中的主要成分,为26个氨基酸组成的碱性多肽。蜂毒疗法是我国利用传统中医中药治疗肿瘤的独特方法。Melittin具有抗菌、抗肿瘤、抗关节炎、抗艾滋病病毒等诸多生物学活性,还具有免疫调节功能,并可对多种肿瘤细胞有直接杀伤作用。白细胞介素-2(interleukin-2,IL-2)具有多种与免疫功能强化有关的生物学活性。其制剂及融合蛋白已有研究报道并取得了一定的效果。

  9. [Regulatory T-cells in systemic lupus erythematosus. IL-2 is decisive for loss of tolerance].

    Science.gov (United States)

    Ohl, K; Tenbrock, K

    2016-04-01

    Systemic lupus erythematosus (SLE) results from loss of immunological tolerance. Regulatory T‑cells (Treg) are major gatekeepers of peripheral tolerance by suppression of autoreactive lymphocytes. Defects in Treg function are therefore possible pathogenetic mechanisms of SLE. Despite this fact published work about numbers and functions of Tregs in SLE are contradictory and the definitive role of Treg in SLE remains unclear. In this review we summarize the current literature about Treg subtypes and the phenotypic markers in human SLE. We also discuss data from mouse models and ex vivo experiments, which provide indications for possible mechanisms that contribute to loss of tolerance. We also discuss the role of interleukin 2 (IL-2), which is decisive for the function of Treg and has been used therapeutically in preliminary trials in human SLE. The identification of novel Treg markers and the development of novel therapeutic approaches, which restore the balance between Treg and autoreactive T‑cells are future goals for research in SLE. PMID:26975190

  10. Altered calcium handling and increased contraction force in human embryonic stem cell derived cardiomyocytes following short term dexamethasone exposure.

    Science.gov (United States)

    Kosmidis, Georgios; Bellin, Milena; Ribeiro, Marcelo C; van Meer, Berend; Ward-van Oostwaard, Dorien; Passier, Robert; Tertoolen, Leon G J; Mummery, Christine L; Casini, Simona

    2015-11-27

    One limitation in using human pluripotent stem cell derived cardiomyocytes (hPSC-CMs) for disease modeling and cardiac safety pharmacology is their immature functional phenotype compared with adult cardiomyocytes. Here, we report that treatment of human embryonic stem cell derived cardiomyocytes (hESC-CMs) with dexamethasone, a synthetic glucocorticoid, activated glucocorticoid signaling which in turn improved their calcium handling properties and contractility. L-type calcium current and action potential properties were not affected by dexamethasone but significantly faster calcium decay, increased forces of contraction and sarcomeric lengths, were observed in hESC-CMs after dexamethasone exposure. Activating the glucocorticoid pathway can thus contribute to mediating hPSC-CMs maturation.

  11. NF-κB Regulates B-Cell-Derived Nerve Growth Factor Expression

    Institute of Scientific and Technical Information of China (English)

    Klaus Heese; Noriko Inoue; Tohru Sawada

    2006-01-01

    In the mammalian brain, four neurotrophins have been identified: nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4/5 (NT-4/5). NGF exerts an important role in the development and functions of the central and peripheral nervous system. However, it has recently been documented that several types of immune cells, such as mast cells, lymphocytes, basophils and eosinophils, produce,store and release NGF. Accumulating preclinical and clinical data indicate that dysfunctions of NGF and the other neurotrophins may contribute to impaired immune responses and concentration of NGF frequently correlates with disease severity. Thus, the aim of this study was to elucidate the potential signaling mechanisms of cytokineneurotrophins interactions contributing to increased NGF levels. Our data show that the transcription factorNF-κB plays a pivotal role in regulating B-cell-derived NGF expression.

  12. Trophoblast lineage cells derived from human induced pluripotent stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Ying, E-mail: ying.chen@hc.msu.edu [Department of Obstetrics, Gynecology and Reproductive Biology, Michigan State University, 333 Bostwick NE, Grand Rapids, MI 49503 (United States); Wang, Kai; Chandramouli, Gadisetti V.R. [Department of Obstetrics, Gynecology and Reproductive Biology, Michigan State University, 333 Bostwick NE, Grand Rapids, MI 49503 (United States); Knott, Jason G. [Developmental Epigenetics Laboratory, Department of Animal Science, Michigan State University (United States); Leach, Richard, E-mail: Richard.leach@hc.msu.edu [Department of Obstetrics, Gynecology and Reproductive Biology, Michigan State University, 333 Bostwick NE, Grand Rapids, MI 49503 (United States); Department of Obstetrics, Gynecology and Women’s Health, Spectrum Health Medical Group (United States)

    2013-07-12

    Highlights: •Epithelial-like phenotype of trophoblast lineage cells derived from human iPS cells. •Trophoblast lineage cells derived from human iPS cells exhibit trophoblast function. •Trophoblasts from iPS cells provides a proof-of-concept in regenerative medicine. -- Abstract: Background: During implantation, the blastocyst trophectoderm attaches to the endometrial epithelium and continues to differentiate into all trophoblast subtypes, which are the major components of a placenta. Aberrant trophoblast proliferation and differentiation are associated with placental diseases. However, due to ethical and practical issues, there is almost no available cell or tissue source to study the molecular mechanism of human trophoblast differentiation, which further becomes a barrier to the study of the pathogenesis of trophoblast-associated diseases of pregnancy. In this study, our goal was to generate a proof-of-concept model for deriving trophoblast lineage cells from induced pluripotency stem (iPS) cells from human fibroblasts. In future studies the generation of trophoblast lineage cells from iPS cells established from patient’s placenta will be extremely useful for studying the pathogenesis of individual trophoblast-associated diseases and for drug testing. Methods and results: Combining iPS cell technology with BMP4 induction, we derived trophoblast lineage cells from human iPS cells. The gene expression profile of these trophoblast lineage cells was distinct from fibroblasts and iPS cells. These cells expressed markers of human trophoblasts. Furthermore, when these cells were differentiated they exhibited invasive capacity and placental hormone secretive capacity, suggesting extravillous trophoblasts and syncytiotrophoblasts. Conclusion: Trophoblast lineage cells can be successfully derived from human iPS cells, which provide a proof-of-concept tool to recapitulate pathogenesis of patient placental trophoblasts in vitro.

  13. Stem cell-derived systems in toxicology assessment.

    Science.gov (United States)

    Suter-Dick, Laura; Alves, Paula M; Blaauboer, Bas J; Bremm, Klaus-Dieter; Brito, Catarina; Coecke, Sandra; Flick, Burkhard; Fowler, Paul; Hescheler, Jürgen; Ingelman-Sundberg, Magnus; Jennings, Paul; Kelm, Jens M; Manou, Irene; Mistry, Pratibha; Moretto, Angelo; Roth, Adrian; Stedman, Donald; van de Water, Bob; Beilmann, Mario

    2015-06-01

    Industrial sectors perform toxicological assessments of their potential products to ensure human safety and to fulfill regulatory requirements. These assessments often involve animal testing, but ethical, cost, and time concerns, together with a ban on it in specific sectors, make appropriate in vitro systems indispensable in toxicology. In this study, we summarize the outcome of an EPAA (European Partnership of Alternatives to Animal Testing)-organized workshop on the use of stem cell-derived (SCD) systems in toxicology, with a focus on industrial applications. SCD systems, in particular, induced pluripotent stem cell-derived, provide physiological cell culture systems of easy access and amenable to a variety of assays. They also present the opportunity to apply the vast repository of existing nonclinical data for the understanding of in vitro to in vivo translation. SCD systems from several toxicologically relevant tissues exist; they generally recapitulate many aspects of physiology and respond to toxicological and pharmacological interventions. However, focused research is necessary to accelerate implementation of SCD systems in an industrial setting and subsequent use of such systems by regulatory authorities. Research is required into the phenotypic characterization of the systems, since methods and protocols for generating terminally differentiated SCD cells are still lacking. Organotypical 3D culture systems in bioreactors and microscale tissue engineering technologies should be fostered, as they promote and maintain differentiation and support coculture systems. They need further development and validation for their successful implementation in toxicity testing in industry. Analytical measures also need to be implemented to enable compound exposure and metabolism measurements for in vitro to in vivo extrapolation. The future of SCD toxicological tests will combine advanced cell culture technologies and biokinetic measurements to support regulatory and

  14. Trophoblast lineage cells derived from human induced pluripotent stem cells

    International Nuclear Information System (INIS)

    Highlights: •Epithelial-like phenotype of trophoblast lineage cells derived from human iPS cells. •Trophoblast lineage cells derived from human iPS cells exhibit trophoblast function. •Trophoblasts from iPS cells provides a proof-of-concept in regenerative medicine. -- Abstract: Background: During implantation, the blastocyst trophectoderm attaches to the endometrial epithelium and continues to differentiate into all trophoblast subtypes, which are the major components of a placenta. Aberrant trophoblast proliferation and differentiation are associated with placental diseases. However, due to ethical and practical issues, there is almost no available cell or tissue source to study the molecular mechanism of human trophoblast differentiation, which further becomes a barrier to the study of the pathogenesis of trophoblast-associated diseases of pregnancy. In this study, our goal was to generate a proof-of-concept model for deriving trophoblast lineage cells from induced pluripotency stem (iPS) cells from human fibroblasts. In future studies the generation of trophoblast lineage cells from iPS cells established from patient’s placenta will be extremely useful for studying the pathogenesis of individual trophoblast-associated diseases and for drug testing. Methods and results: Combining iPS cell technology with BMP4 induction, we derived trophoblast lineage cells from human iPS cells. The gene expression profile of these trophoblast lineage cells was distinct from fibroblasts and iPS cells. These cells expressed markers of human trophoblasts. Furthermore, when these cells were differentiated they exhibited invasive capacity and placental hormone secretive capacity, suggesting extravillous trophoblasts and syncytiotrophoblasts. Conclusion: Trophoblast lineage cells can be successfully derived from human iPS cells, which provide a proof-of-concept tool to recapitulate pathogenesis of patient placental trophoblasts in vitro

  15. Detection of T lymphocyte subsets and mIL-2R on surface of PBMC in patients with hepatitis B

    OpenAIRE

    Wang, Ke-Xia; Peng, Jiang-Long; Wang, Xue-Feng; Tian, Ye; Wang, Jian; Li, Chao-Pin

    2003-01-01

    AIM: To study the levels of T lymphocyte subsets and membrane interleukin-2 receptor (mIL-2R) on surface of peripheral blood mononuclear cells (PBMCs) of patients with hepatitis B and its role in the pathogenesis of hepatitis B.

  16. Shock Waves Increase T-cell Proliferation or IL-2 Expression by Activating p38 MAP Kinase

    Institute of Scientific and Technical Information of China (English)

    Tie-Cheng YU; Yi LIU; Yan TAN; Yanfang JIANG; Xueqing ZHENG; Xinxiang XU

    2004-01-01

    Shock waves were elicited by transient pressure disturbances, which could be used to treat musculoskeletal disorders. In present studies, we i. nvestigated whether the low-density shock waves (LDSWs), which are able to damage plasma membrane without impairing the vimentin or other organelles, might augment T-cell proliferation as well as IL-2 expression, and if mitogen activated protein kinase p38 (p38 MAPK)might be an underlying mechanism through which the LDSWs enhanced T-cell function. We found that the LDSWs increased activation of p38 MAPK in Jurkat T cells. The LDSWs alone didn't result in the T-cell proliferation and IL-2 expression. However, in combination with other stimuli, LDSWs could augment the T-cell proliferation and IL-2 expression. Inhibition of p38 MAPK using SB203580 reduced the stimulatory effects of the LDSWs, which indicated that the LDSWs enhanced IL-2 expression through a mechanism that involved p38 MAPK activation. We concluded that the p38 MAPK activation played a key role in the regulation of T cell function by the LDSWs.

  17. High-Dose Interleukin-2 (HD IL-2 Therapy Should Be Considered for Treatment of Patients with Melanoma Brain Metastases

    Directory of Open Access Journals (Sweden)

    Melinda B. Chu

    2013-01-01

    Full Text Available A retrospective review was performed on patients with stable melanoma brain metastases treated with HD IL-2 therapy (720,000 IU/kg per dose intravenously; 14 doses, 2 cycles per course, maximum 2 courses from January 1999 to June 2011 at Saint Louis University. There were 5 men and 3 women; median age was 52.2 years (26.8–61.1 years. One patient started treatment with lung lesions only (after resection of melanoma brain disease and experienced partial response. Seven patients had brain metastases at treatment initiation. Median overall survival (mOS for entire cohort ( was 8.7 months (2.1 to 19.0 months. All patients with brain metastases at first dose ( showed progressive disease; mOS was 6.7 months (range 2.1–18.2 months for this group. Patients received radiosurgery and whole brain radiation before and after HD IL-2 therapy. One patient had symptoms suggestive of neurotoxicity. A history of alcohol abuse was revealed during admission. The patient's symptoms improved with initiation of an alcohol withdrawal protocol. In this analysis, patients with melanoma brain metastases received HD IL-2 without treatment-related mortality. We think that HD IL-2 should be considered as a treatment option in patients with melanoma brain metastases who are otherwise eligible for therapy.

  18. Induction of prolonged survival of CD4+ T lymphocytes by intermittent IL-2 therapy in HIV-infected patients

    Science.gov (United States)

    Kovacs, Joseph A.; Lempicki, Richard A.; Sidorov, Igor A.; Adelsberger, Joseph W.; Sereti, Irini; Sachau, William; Kelly, Grace; Metcalf, Julia A.; Davey, Richard T.; Falloon, Judith; Polis, Michael A.; Tavel, Jorge; Stevens, Randy; Lambert, Laurie; Hosack, Douglas A.; Bosche, Marjorie; Issaq, Haleem J.; Fox, Stephen D.; Leitman, Susan; Baseler, Michael W.; Masur, Henry; Di Mascio, Michele; Dimitrov, Dimiter S.; Lane, H. Clifford

    2005-01-01

    HIV infection leads to decreases in the number of CD4+ T lymphocytes and an increased risk for opportunistic infections and neoplasms. The administration of intermittent cycles of IL-2 to HIV-infected patients can lead to profound increases (often greater than 100%) in CD4 cell number and percentage. Using in vivo labeling with 2H-glucose and BrdU, we have been able to demonstrate that, although therapy with IL-2 leads to high levels of proliferation of CD4 as well as CD8 lymphocytes, it is a remarkable preferential increase in survival of CD4 cells (with half-lives that can exceed 3 years) that is critical to the sustained expansion of these cells. This increased survival was time-dependent: the median half-life, as determined by semiempirical modeling, of labeled CD4 cells in 6 patients increased from 1.7 weeks following an early IL-2 cycle to 28.7 weeks following a later cycle, while CD8 cells showed no change in the median half-life. Examination of lymphocyte subsets demonstrated that phenotypically naive (CD27+CD45RO–) as well as central memory (CD27+CD45RO+) CD4 cells were preferentially expanded, suggesting that IL-2 can help maintain cells important for host defense against new antigens as well as for long-term memory to opportunistic pathogens. PMID:16025158

  19. Spontaneous resolution of acute rejection and tolerance induction with IL-2 fusion protein in vascularized composite allotransplantation.

    Science.gov (United States)

    Jindal, R; Unadkat, J; Zhang, W; Zhang, D; Ng, T W; Wang, Y; Jiang, J; Lakkis, F; Rubin, P; Lee, W P A; Gorantla, V S; Zheng, X X

    2015-05-01

    Vascularized composite allotransplantation (VCA) has emerged as a treatment option for treating nonlife-threatening conditions. Therefore, in order to make VCA a safe reconstruction option, there is a need to minimize immunosuppression, develop tolerance-inducing strategies and elucidate the mechanisms of VCA rejection and tolerance. In this study we explored the effects of hIL-2/Fc (a long-lasting human IL-2 fusion protein), in combination with antilymphocyte serum (ALS) and short-term cyclosporine A (CsA), on graft survival, regulatory T cell (Treg) proliferation and tolerance induction in a rat hind-limb transplant model. We demonstrate that hIL-2/Fc therapy tips the immune balance, increasing Treg proliferation and suppressing effector T cells, and permits VCA tolerance as demonstrated by long-term allograft survival and donor-antigen acceptance. Moreover, we observe two distinct types of acute rejection (AR), progressive and reversible, within hIL-2/Fc plus ALS and CsA treated recipients. Our study shows differential gene expression profiles of FoxP3 versus GzmB, Prf1 or interferon-γ in these two types of AR, with reversible rejection demonstrating higher Treg to Teff gene expression. This correlation of gene expression profile at the first clinical sign of AR with VCA outcomes can provide the basis for further inquiry into the mechanistic aspects of VCA rejection and future drug targets. PMID:25676865

  20. Paclitaxel enhances therapeutic efficacy of the F8-IL2 immunocytokine to EDA-fibronectin-positive metastatic human melanoma xenografts.

    Science.gov (United States)

    Moschetta, Michele; Pretto, Francesca; Berndt, Alexander; Galler, Kerstin; Richter, Petra; Bassi, Andrea; Oliva, Paolo; Micotti, Edoardo; Valbusa, Giovanni; Schwager, Kathrin; Kaspar, Manuela; Trachsel, Eveline; Kosmehl, Hartwig; Bani, Maria Rosa; Neri, Dario; Giavazzi, Raffaella

    2012-04-01

    The selective delivery of bioactive agents to tumors reduces toxicity and enhances the efficacy of anticancer therapies. In this study, we show that the antibody F8, which recognizes perivascular and stromal EDA-fibronectin (EDA-Fn), when conjugated to interleukin-2 (F8-IL2) can effectively inhibit the growth of EDA-Fn-expressing melanomas in combination with paclitaxel. We obtained curative effects with paclitaxel administered before the immunocytokine. Coadministration of paclitaxel increased the uptake of F8 in xenografted melanomas, enhancing tumor perfusion and permeability. Paclitaxel also boosted the recruitment of F8-IL2-induced natural killer (NK) cells to the tumor, suggesting a host response as part of the observed therapeutic benefit. In support of this likelihood, NK cell depletion impaired the antitumor effect of paclitaxel plus F8-IL2. Importantly, this combination reduced both the tumor burden and the number of pulmonary metastatic nodules. The combination did not cause cumulative toxicity. Together, our findings offer a preclinical proof that by acting on the tumor stroma paclitaxel potentiates the antitumor activity elicited by a targeted delivery of IL2, thereby supporting the use of immunochemotherapy in the treatment of metastatic melanoma. PMID:22392081

  1. Identical genetic control of MLC reactivity to different MHC incompatibilities, independent of production of and response to IL-2

    Energy Technology Data Exchange (ETDEWEB)

    Holan, V.; Lipoldova, M.; Demant, P. [Inst. of Molecular Genetics, Prague (Czech Republic)

    1996-12-31

    The inbred strain STS/A exhibits a higher proliferative response in the mixed lymphocyte culture (MLC) to stimulator cells of all 11 tested inbred mouse strains with 10 different major histocompatibility complex (MHC) haplotypes, as well as to stimulation with IL-2 than does the strain BALB/cHeA. However, alloantigen-stimulated BALB/c cells produce more IL-2 than STS/A cells. To study the genetic basis of these differences, we used 20 recombinant congenic strains (RCS) of the CcS/Dem series. Each of these CcS/Dem RC strains contains a different subset of about 12.5% of genes from the STS/A strain and the remaining approximately 87.5% of BALB/c origin genes. As a result the multiple non-linked genes responsible for phenotypic differences between BALB/c and STS/A became separated into different CcS/Dem strains. The strain distribution pattern (SDP) of high or low MLC response of individual CcS/Dem strains to stimulator cells of four different strains was almost identical, indicating that differences in responsiveness to different alloantigens is largely controlled by the same genes. The SDP of IL-2 stimulation was different from that of MLC responsiveness. The differences in the proliferative responses observed among individual CcS/Dem strains were not due to differences in numbers of CD3{sup +}, CD4{sup +} or CD8{sup +} cells or to the observed differences in IL-2 production, and hence they likely reflect genetically determined intrinsic properties of T cells. These results show that a set of non-linked genes controls proliferative responses in MLC irrespective of the MHC haplotype of the stimulator cells, and that stimulation with IL-2 and production of IL-2 are controlled by different subsets of genes. Since the genomes of all RCS are extensively characterized by microsatellite markers, they can be used to map the genes controlling proliferative responsiveness to stimulation with alloantigens and IL-2. 33 refs., 9 figs.

  2. Lymphokine-activated killer cell phenomenon. III. Evidence that IL-2 is sufficient for direct activation of peripheral blood lymphocytes into lymphokine-activated killer cells

    OpenAIRE

    1983-01-01

    Purified interleukin 2 (IL-2) was found to be sufficient for direct activation of peripheral blood lymphocytes into lymphokine-activated killer (LAK) cells. The LAK activation factor was directly and consistently associated with IL-2 activity using classic protein purification techniques, adsorption to IL-2-dependent cell lines, and inhibition with anti-Tac antibody. As yet, no other cytokines have been found that perform the same role.

  3. Engraftment of human HSCs in nonirradiated newborn NOD-scid IL2rγnull mice is enhanced by transgenic expression of membrane-bound human SCF

    OpenAIRE

    Brehm, Michael A.; Racki, Waldemar J.; Leif, Jean; Burzenski, Lisa; Hosur, Vishnu; Wetmore, Amber; Gott, Bruce; Herlihy, Mary; Ignotz, Ronald; Dunn, Raymond; Shultz, Leonard D.; Greiner, Dale L.

    2012-01-01

    Immunodeficient mice engrafted with human HSCs support multidisciplinary translational experimentation, including the study of human hematopoiesis. Heightened levels of human HSC engraftment are observed in immunodeficient mice expressing mutations in the IL2-receptor common γ chain (IL2rg) gene, including NOD-scid IL2rγnull (NSG) mice. Engraftment of human HSC requires preconditioning of immunodeficient recipients, usually with irradiation. Such preconditioning increases the expression of st...

  4. Genetic variants in IL2RA and IL7R affect multiple sclerosis disease risk and progression

    Science.gov (United States)

    Traboulsee, Anthony L.; Bernales, Cecily Q.; Ross, Jay P.; Lee, Joshua D.; Sadovnick, A. Dessa; Vilariño-Güell, Carles

    2016-01-01

    Multiple sclerosis (MS) is a common demyelinating neurodegenerative disease with a strong genetic component. Previous studies have associated genetic variants in IL2RA and IL7R in the pathophysiology of the disease. In this study we describe the association between IL2RA (rs2104286) and IL7R (rs6897932) in the Canadian population. Genotyping 1,978 MS patients and 830 controls failed to identify any significant association between these variants and disease risk. However, stratified analysis for family history of disease, and disease course identified a trend towards association for IL2RA in patients without a family history (p = 0.05; odds ratio = 0.77), and a significant association between IL7R and patients who developed progressive MS (PrMS) (p = 0.002; odds ratio = 0.73). Although not statistically significant, the effect of IL2RA (rs2104286) in patients without a family history of MS indicates that the genetic components for familial and sporadic disease are perhaps distinct. This data suggests the onset of sporadic disease is likely determined by a large number of variants of small effect, whereas MS in patients with a family history of disease is caused by a few deleterious variants. In addition, the significant association between PrMS and rs6897932 indicates that IL7R may not be disease-causing but a determinant of disease course. Further characterization of the effect of IL2RA and IL7R genetic variants in defined MS subtypes is warranted to evaluate the effect of these genes on specific clinical outcomes and to further elucidate the mechanisms of disease onset and progression. PMID:24770783

  5. CONSTRUCTION AND EXPRESSION OF ADENOASSOCIATED VIRUS- BASED PLASMID EXPRESSING VECTORS CONTAINING hIL- 2 GENE OR mIFN-γ GENE

    Institute of Scientific and Technical Information of China (English)

    张景迎; 梁宏立; 陈诗书

    2000-01-01

    Objective To improve the plasmid vectors in gene therapy, adeno - associated virus (AA V) based plasmid expressing vectors containing hIL-2 gene or mIFN-γ gene were constructed and its expression in transfected cells was studied. Methods By means of step to step cloning, promoter CMVp was placed at the downstream of 5' inverted terminal repeat from AA V (AA V- ITR) of pAP, hIL- 2 gene or mIFN- γ gene inserted into pAC between CMVp and poly A. Then intron A was inserted into pAC- hIL - 2 or pAC- mIFN- γ between CMVp and IL - 2 gene or IFNγ gene to construct pAI- hIL - 2 or pAI- mIFN - γ. Liposome -plasmid complexes were formed by mixing Dosper with these AAV-based plasmids containing hIL-2 gene or mIFN-γgene. Results High biological activities of IL - 2 or IFN- γ could be detected in the supernatants of NIH3T3 and MM45T. Li cells after transfection. Insertion of intron A into pAC-hIL-2 or pAC-mIFN-γ improved the expression of IL- 2 or IFN- γ. Conclusion These data demonstrated that the constructed AA V- based plasmid expressing vectors could efficiently express therapeutic genes in cultured cells and could be used as a nonviral gene transfer system in human gene therapy.

  6. Defective production of interleukin 2 in patients with Chagas' disease: purified IL-2 augments in vitro response in patients with chagasic cardiomyopathy

    Directory of Open Access Journals (Sweden)

    Luis Briceno

    1996-10-01

    Full Text Available The production of interleukin 2 (IL-2 by peripheral blood mononuclear cells, from patients with different clinical forms of Chagas disease and healthy controls, was evaluated after stimulation with Trypanosoma cruzi antigen, PPD and PHA. PHA induced higher production of IL-2 in infected patients than healthy controls. No diferences were found between infected groups. With PPD the trend was similar, the only difference was that asymptomatic infected patients (INF showed higher levels of IL-2 production than patients with cardiomyopathy (CDM. With T. cruzi antigen, most patients showed little or no IL-2 production at 24 hr, a peak at 48 hr and an abrupt fall at 72 hr. A similar pattern of IL- 2 production was observed in INF and CDM. To evaluate the physiologic relevance of the deficit in IL-2 production, we studied the effect of non-mitogenic concentratios of IL-2 in the proliferative response to specific antigens. The addition of IL-2 only enhanced the proliferative response of CDM patients. These observations suggest that patients suffering Chagas' disease, particularly CDM, have a significant reduction in the capacity to produce IL-2. These findings could be of importance in the pathogenesis of Chagas' disease.

  7. Down-regulation of Runx1 Expression by TCR Signal Involves an Autoregulatory Mechanism and Contributes to IL-2 Production*

    OpenAIRE

    Wong, Won Fen; Kurokawa, Mineo; Satake, Masanobu; Kohu, Kazuyoshi

    2011-01-01

    Runx1 transcription factor plays multiple roles in T cell development, differentiation, and function. However, the regulatory mechanisms and functional significance of high Runx1 protein expression in resting peripheral CD4+ T cells is not well understood. Here, we demonstrate that T-cell receptor (TCR) activation down-regulates distal Runx1 transcription, resulting in a significant reduction of Runx1 protein. Interestingly, this down-regulation of distal Runx1 transcription appears to be med...

  8. Correlation of bone marrow stromal cell-derived exosomes with Hedgehog signaling in the progress of breast cancer%骨髓间充质干细胞源性外泌体与Hedgehog信号通路在乳腺癌发展中的关系研究

    Institute of Scientific and Technical Information of China (English)

    王丹丹; 陈建中; 亢春彦

    2015-01-01

    目的:研究Hedgehog信号通路在骨髓间充质干细胞( BMSCs)来源exosome介导小鼠4T1乳腺癌细胞发展过程中的作用。方法使用梯度离心分离法、差速贴壁培养方法及超速离心法分离C57BL/6小鼠BMSCs及其exosome,使用MTT法、细胞划痕实验及western blot技术分析exosome干预前后4T1癌细胞增殖、迁移和侵袭能力以及Hedgehog信号通路相关蛋白表达情况,之后使用GANT61(Hedgehog信号通路阻断剂)验证Hedgehog信号通路在BM-SCs来源的exosome介导乳腺癌细胞增殖、迁移和侵袭过程中的作用。结果 BMSCs来源的exosome显著上调了4T1细胞Hedgehog信号通路,exosome显著增加了4T1细胞的增殖及迁移、侵袭能力,而这一作用被GANT61所削减。结论 BMSCs来源的exosome能够通过上调Hedgehog信号通路增加4T1小鼠乳腺癌细胞的增殖、迁移及侵袭能力。%Objective To investigate the roles of Hedgehog signaling pathway in the BMSCs derived exosome-in-duced progression of 4T1 mouse breast cancer cells. Methods Prepare the BMSCs and exosome by gradient centrifugation separation, differential adhesion method and ultracentrifugation method;use the MTT test, cell scratch test and western blot to examine the effect of BMSCs-exosome on the proliferation, migration and invasion ability of 4T1 cells, then use the GANT61, an inhibitor of Hedgehog signaling pathway, to test the mechanism of exosome-induced changes in the 4T1 cells. Results BMSCs-exosome significantly up-regulated the Hedgehog signaling pathway of the 4T1 cells. The exosome signifi-cantly increased the proliferation, migaration and invasion ability of 4T1 cells in vitro, and this effect was abolished by GANT61. Conclusions BMSCs derived exosome can promote the proliferation, migration and invasion ability by upregu-late Hedgehog signaling pathway.

  9. IL-2 or IL-4 mRNA as a potential flow cytometric marker molecule for selective collection of living T helper 1 or T helper 2 lymphocytes.

    Science.gov (United States)

    Ishibashi, Kaname; Tsuji, Akihiko

    2003-06-01

    Flow cytometry has been widely used to analyze and sort out particular types of living cells that have specific marker molecules. In many cases, marker proteins are present on the cell surface and are detected by monoclonal antibodies against them. However, there are some cases in which cells do not have specific marker molecules on their surface. In this situation, it would be useful if mRNA that is expressed specifically in the particular cell could be used as a marker molecule. We previously reported that mRNA can be detected in living cells by hybridizing a pair of fluoreophore (donor or acceptor)-labeled oligonucleotides to adjacent locations on the target mRNA in the cytoplasm of cells (Tsuji, A.; Koshimoto, H.; Sato, Y.; Hirano, M.; Sei-Iida, Y.; Kondo, S.; Ishibashi, K. Biophys. J. 2000, 78, 3260-3274). On the formed hybrid of the two fluorescent oligonucleotides with the target mRNA, the distance between the two fluorophores becomes very close, which results in fluorescence resonance energy transfer (FRET). Combining this fluorescent labeling method for mRNA with flow cytometry, we have examined the isolation of living CD4+ T helper lymphocytes expressing IL-2 mRNA (Th1) or IL-4 mRNA (Th2). A pair of fluorescent oligonucleotides for hybridizing to IL-2 or IL-4 mRNA were introduced into activated CD4+ T lymphocytes by electroporation. The cells were applied to FACS and analyzed by FRET signals. Th1 or Th2 lymphocytes were exclusively sorted from their mixed populations in activated CD4+ T cells. Our results demonstrate that it is possible to use mRNA as marker molecules to analyze and isolate living cells in flow cytometry. PMID:12948141

  10. Large-scale generation of cell-derived nanovesicles

    Science.gov (United States)

    Jo, W.; Kim, J.; Yoon, J.; Jeong, D.; Cho, S.; Jeong, H.; Yoon, Y. J.; Kim, S. C.; Gho, Y. S.; Park, J.

    2014-09-01

    Exosomes are enclosed compartments that are released from cells and that can transport biological contents for the purpose of intercellular communications. Research into exosomes is hindered by their rarity. In this article, we introduce a device that uses centrifugal force and a filter with micro-sized pores to generate a large quantity of cell-derived nanovesicles. The device has a simple polycarbonate structure to hold the filter, and operates in a common centrifuge. Nanovesicles are similar in size and membrane structure to exosomes. Nanovesicles contain intracellular RNAs ranging from microRNA to mRNA, intracellular proteins, and plasma membrane proteins. The quantity of nanovesicles produced using the device is 250 times the quantity of naturally secreted exosomes. Also, the quantity of intracellular contents in nanovesicles is twice that in exosomes. Nanovesicles generated from murine embryonic stem cells can transfer RNAs to target cells. Therefore, this novel device and the nanovesicles that it generates are expected to be used in exosome-related research, and can be applied in various applications such as drug delivery and cell-based therapy.

  11. Sodium arsenite-induced inhibition of cell proliferation is related to inhibition of IL-2 mRNA expression in mouse activated T cells

    Energy Technology Data Exchange (ETDEWEB)

    Conde, Patricia; Acosta-Saavedra, Leonor C.; Calderon-Aranda, Emma S. [Centro de Investigacion y de Estudios Avanzados, CINVESTAV, Seccion Toxicologia, P.O. Box 14-740, Mexico, D.F. (Mexico); Goytia-Acevedo, Raquel C. [Universidad Juarez del Estado de Durango, Facultad de Medicina, Gomez Palacio, Durango (Mexico)

    2007-04-15

    A proposed mechanism for the As-induced inhibition of cell proliferation is the inhibition of IL-2 secretion. However, the effects of arsenite on IL-2 mRNA expression or on the ERK pathway in activated-T cells have not yet been described. We examined the effect of arsenite on IL-2 mRNA expression, cell activation and proliferation in PHA-stimulated murine lymphocytes. Arsenite (1 and 10 {mu}M) decreased IL-2 mRNA expression, IL-2 secretion and cell proliferation. Arsenite (10 {mu}M) strongly inhibited ERK-phosphorylation. However, the partial inhibition (50%) of IL-2 mRNA produced by 1 {mu}M, consistent with the effects on IL-2 secretion and cell proliferation, could not be explained by the inhibition of ERK-phosphorylation, which was not affected at this concentration. The inhibition of IL-2 mRNA expression caused by 1 {mu}M could be associated to effects on pathways located downstream or parallel to ERK. Arsenite also decreased early activation (surface CD69{sup +} expression) in both CD4{sup +} and CD8{sup +}, and decreased total CD8{sup +} count without significantly affecting CD4{sup +}, supporting that the cellular immune response mediated by cytotoxic T cells is an arsenic target. Thus, our results suggest that arsenite decreases IL-2 mRNA levels and T-cell activation and proliferation. However, further studies on the effects of arsenite on IL-2 gene transcription and IL-2 mRNA stability are needed. (orig.)

  12. Improved flow cytometric assessment reveals distinct microvesicle (cell-derived microparticle signatures in joint diseases.

    Directory of Open Access Journals (Sweden)

    Bence György

    Full Text Available INTRODUCTION: Microvesicles (MVs, earlier referred to as microparticles, represent a major type of extracellular vesicles currently considered as novel biomarkers in various clinical settings such as autoimmune disorders. However, the analysis of MVs in body fluids has not been fully standardized yet, and there are numerous pitfalls that hinder the correct assessment of these structures. METHODS: In this study, we analyzed synovial fluid (SF samples of patients with osteoarthritis (OA, rheumatoid arthritis (RA and juvenile idiopathic arthritis (JIA. To assess factors that may confound MV detection in joint diseases, we used electron microscopy (EM, Nanoparticle Tracking Analysis (NTA and mass spectrometry (MS. For flow cytometry, a method commonly used for phenotyping and enumeration of MVs, we combined recent advances in the field, and used a novel approach of differential detergent lysis for the exclusion of MV-mimicking non-vesicular signals. RESULTS: EM and NTA showed that substantial amounts of particles other than MVs were present in SF samples. Beyond known MV-associated proteins, MS analysis also revealed abundant plasma- and immune complex-related proteins in MV preparations. Applying improved flow cytometric analysis, we demonstrate for the first time that CD3(+ and CD8(+ T-cell derived SF MVs are highly elevated in patients with RA compared to OA patients (p=0.027 and p=0.009, respectively, after Bonferroni corrections. In JIA, we identified reduced numbers of B cell-derived MVs (p=0.009, after Bonferroni correction. CONCLUSIONS: Our results suggest that improved flow cytometric assessment of MVs facilitates the detection of previously unrecognized disease-associated vesicular signatures.

  13. Purification and Biological Activity of Recombinant Melittin-88 ArgIL-2%重组蜂毒素-基因变构IL-2的纯化与生物活性

    Institute of Scientific and Technical Information of China (English)

    刘明军; 王斌; 钱冬萌; 丁守怡; 闫志勇; 宋旭霞

    2006-01-01

    目的 制备纯化重组蜂毒素-基因变构IL-2(Melittin-88 ArgIL-2)嵌合蛋白,并检测其生物活性.方法 将含表达载体的大肠杆菌DH5α,经IPTG诱导表达.表达产物经离子交换层析与亲和层析进行纯化,SDS-PAGE鉴定,MTT染色法检测嵌合蛋白对Hela细胞增殖的抑制作用.结果 所表达的可溶性蛋白,纯化后纯度达95%,蛋白浓度0.5 g/L.纯化后的嵌合蛋白在体外能抑制Hela细胞的增殖.结论 已成功地制备并纯化了重组melittin-88 ArgIL-2嵌合蛋白,该蛋白具有生物活性,为中试放大和纯化工艺研究奠定了基础.

  14. Low prevalence of antibodies and other plasma factors binding to CC chemokines and IL-2 in HIV-positive patients

    DEFF Research Database (Denmark)

    Meyer, C N; Svenson, M; Schade Larsen, C;

    2000-01-01

    Neutralizing cytokine antibodies are found in healthy and diseased individuals, including patients treated with recombinant cytokines. Identification of CCR-5 as co-receptor for HIV has focused interest on CC chemokines and their potential therapeutic use. Chemokine-binding components in plasma...... of HIV-infected patients were therefore assessed by radioimmunoassay and radioreceptor assay. IgG from 4/505 HIV patients and 9/2000 healthy controls (p>0.05) bound rMIP-1alpha and rMIP-1beta, but not rRANTES. No other plasma factors bound the chemokines. The antibodies inhibited receptor binding of both...... chemokines. There was no association between presence of antibodies and disease stage or HIV progression rate. Three of 11 patients treated with rIL-2 developed IgG antibodies suppressing cellular binding and growth promotion of rIL-2. Hence, circulating factors, including antibodies MIP-1alpha/MIP-1beta...

  15. Effects of Warm Needling at Zusanli (ST 36) on NO and IL-2 Levels in the Middle-Aged and Old People

    Institute of Scientific and Technical Information of China (English)

    李苏; 陈开阳; 吴英; 焦健慧; 陶立富

    2003-01-01

    42 middle-aged and old people at the age between 55-70 years were selected and given the warm needling at Zusanli (ST 36), and their IL-2 and NO contents of peripheral blood before and after acupuncture were determined. The results showed that IL-2 and NO contents increased significantly after the warm needling (P<0.01).

  16. TK gene combined with mIL-2 and mGM-CSF genes in treatment of gastric cancer

    Institute of Scientific and Technical Information of China (English)

    Shan-Yu Guo; Qin-Long Gu; Zheng-Gang Zhu; He-Qun Hong; Yan-Zhen Lin

    2003-01-01

    AIM: Cancer gene therapy has received more and moreattentions in the recent decade. Various systems of genetherapy for cancer have been developed. One of the mostpromising choices is the suicide gene. The product ofthymidine kinase (TK) gene can convert ganciclovir (GCV)to phosphorylated GCV, which inhibits the synthesis of cellDNA, and then induces the cells to death. Cytolines play animportant role in anti-tumor immunity. This experiment wasdesigned to combine theTK gene and mIL-2/mGM-CSFgenes to treat gastric cancer, and was expected to producea marked anti-tumor effect.METHODS: TK gene was constructed into the retroviralvector pLxSN, and the mIL-2 and mGM-CSF genes wereinserted into the eukaryotic expressing vector pIRES. Thegastric cancer cells were transfected by retroviral serum thatwas harvested from the package cells. In vitro study, thetransfected gastric cancer cells were maintained in the GCV-contained medium, to assay the cell killing effect andbystander effect. In vivo experiment, retroviral serum andcytokines plasmid were transfected into tumor-bearing mice,to observe the changes of tumor volumes and survival ofthe mice.RESULTS: In vitro experiment, 20 % TK gene transducedcells could cause 70-80 % of total cells to death. In vivoresults showed that there was no treatment effect in controlgroup and TK/GCV could inhibit the tumor growth. Thestrongest anti-tumor effect was shown in TK+mIL-2+mGM-CSF group. The pathologic examination showed necrosis ofthe cancer in the treated groups.CONCLUSION: TK/GCV can kill tumor cells and inhibit thetumor growth in vivo IL-2 and GM-CSF strongly enhancethe anti-tumor effect. Through the retrovirus and liposomemethods, the suicide gene and cytokine genes are allexpressed in the tissues.

  17. Dendritic Cell-Derived Exosomes Stimulate Stronger CD8+ CTL Responses and Antitumor Immunity than Tumor Cell-Derived Exosomes

    Institute of Scientific and Technical Information of China (English)

    Siguo Hao; Ou Bai; Jinying Yuan; Mabood Qureshi; Jim Xiang

    2006-01-01

    Exosomes (EXO) derived from dendritic cells (DC) and tumor cells have been used to stimulate antitumor immune responses in animal models and in clinical trials. However, there has been no side-by-side comparison of the stimulatory efficiency of the antitumor immune responses induced by these two commonly used EXO vaccines. In this study, we selected to study the phenotype characteristics of EXO derived from a transfected EG7 tumor cells expressing ovalbumin (OVA) and OVA-pulsed DC by flow cytometry. We compared the stimulatory effect in induction of OVA-specific immune responses between these two types of EXO. We found that OVA protein-pulsed DCovA-derived EXO (EXODC) can more efficiently stimulate naive OVA-specific CD8+ T cell proliferation and differentiation into cytotoxic T lymphocytes in vivo, and induce more efficient antitumor immunity than EG7 tumor cell-derived EXO (EXOEG7). In addition, we elucidated the important role of the host DC in EXO vaccines that the stimulatory effect of EXO is delivered to T cell responses by the host DC. Therefore, DC-derived EXO may represent a more effective EXO-based vaccine in induction of antitumor immunity.

  18. Short-term intratracheal use of PEG-modified IL-2 and glucocorticoid persistently alleviates asthma in a mouse model

    Science.gov (United States)

    Wu, Kefei; Ma, Jiexian; Bai, Weiya; Cui, Xiaoxian; Han, Tao; Wang, Shiyuan; Xie, Youhua; Xie, Yanhui

    2016-01-01

    Regulatory T (Treg) cells play an important role in allergic airway diseases, and upregulation of Treg cells is a potential therapeutic strategy for asthma. In this study, we show that short-term intratracheal use of IL-2 combined with glucocorticoid alleviates antigen-induced airway inflammation and reduces airway hyperresponsiveness by expanding antigen-nonspecific Treg cells, with a decrease in T helper 2 (Th2) cells and Th2-associated cytokines. We also designed a long-acting polyethylene glycol (PEG)-modified IL-2 and demonstrated that the optimal dosage form is IL-2(PEG) plus budesonide, which can upregulate Treg cells and ameliorate asthma at a lower dose. The therapeutic effect was faster than treatment with dexamethasone and was effective at a low dose suitable for humans that could last for at least 6 weeks. This study unveils a new therapeutic regimen and suggests that such endogenous Treg therapy could be a useful tool to persistently alleviate asthma. PMID:27527926

  19. Short-term intratracheal use of PEG-modified IL-2 and glucocorticoid persistently alleviates asthma in a mouse model.

    Science.gov (United States)

    Wu, Kefei; Ma, Jiexian; Bai, Weiya; Cui, Xiaoxian; Han, Tao; Wang, Shiyuan; Xie, Youhua; Xie, Yanhui

    2016-01-01

    Regulatory T (Treg) cells play an important role in allergic airway diseases, and upregulation of Treg cells is a potential therapeutic strategy for asthma. In this study, we show that short-term intratracheal use of IL-2 combined with glucocorticoid alleviates antigen-induced airway inflammation and reduces airway hyperresponsiveness by expanding antigen-nonspecific Treg cells, with a decrease in T helper 2 (Th2) cells and Th2-associated cytokines. We also designed a long-acting polyethylene glycol (PEG)-modified IL-2 and demonstrated that the optimal dosage form is IL-2(PEG) plus budesonide, which can upregulate Treg cells and ameliorate asthma at a lower dose. The therapeutic effect was faster than treatment with dexamethasone and was effective at a low dose suitable for humans that could last for at least 6 weeks. This study unveils a new therapeutic regimen and suggests that such endogenous Treg therapy could be a useful tool to persistently alleviate asthma. PMID:27527926

  20. Recovery and Biodistribution of Ex Vivo Expanded Human Erythroblasts Injected into NOD/SCID/IL2Rγnull mice

    Directory of Open Access Journals (Sweden)

    Barbara Ghinassi

    2011-01-01

    Full Text Available Ex vivo expanded erythroblasts (EBs may serve as advanced transfusion products provided that lodgment occurs in the macrophage-niche of the marrow permitting maturation. EBs expanded from adult and cord blood expressed the receptors (CXCR4, VLA-4, and P-selectin ligand 1 necessary for interaction with macrophages. However, 4-days following transfusion to intact NOD/SCID/IL2Rγnull mice, CD235apos EBs were observed inside CD235aneg splenic cells suggesting that they underwent phagocytosis. When splenectomized and intact NOD/SCID/IL2Rγnull mice were transfused using retrovirally labeled human EBs, human cells were visualized by bioluminescence imaging only in splenectomized animals. Four days after injection, human CD235apos cells were detected in marrow and liver of splenectomized mice but only in spleen of controls. Human CD235apos erythrocytes in blood remained low in all cases. These studies establish splenectomized NOD/SCID/IL2Rγnull mice as a suitable model for tracking and quantification of human EBs in vivo.

  1. Effects of flurbiprofen axetil on postoperative serum IL-2 and IL-6 levels in patients with colorectal cancer.

    Science.gov (United States)

    Jiang, W W; Wang, Q H; Peng, P; Liao, Y J; Duan, H X; Xu, M; Li, Y; Zhang, P B

    2015-01-01

    We explored the effects of flurbiprofen axetil on interleukin (IL)-2 and IL-6 levels in postoperative patients with colorectal cancer. A total of 120 patients (American Society of Anesthesiologists I and II) scheduled to undergo colorectal cancer surgery were randomly divided into 3 groups (N = 40 in each group): flurbiprofen axetil group (group F), morphine group (group M), and tramadol group (group T). Group M received 0.1 mg/kg morphine, group T received 1.5 mg/kg tramadol, and group F received 1.5 mg/kg flurbiprofen axetil. Patients in the 3 groups were administered treatments through intravenous injection 10 min before surgery. Serum IL-2 and IL-6 levels were detected. Postoperative adverse reactions were recorded, such as nausea, vomiting, and pruritus. The serum IL-6 level of the 3 groups increased 3 h after surgery. Compared with group M, IL-6 level was higher in group T and group F at 1 day after the surgery, and the differences between group M and the other groups were significant (P < 0.05). Moreover, the incidence of adverse reactions was significantly different among 3 groups (P < 0.05). Flurbiprofen axetil promoted the secretion of IL-2 and inhibited IL-6; additionally, flurbiprofen axetil may have a lower incidence of adverse reactions compared to other treatments.

  2. Long-Lasting Complete Responses in Patients with Metastatic Melanoma after Adoptive Cell Therapy with Tumor-Infiltrating Lymphocytes and an Attenuated IL2 Regimen

    DEFF Research Database (Denmark)

    Andersen, Rikke; Donia, Marco; Ellebaek, Eva;

    2016-01-01

    administered together with TILs are severe. To further scrutinize whether similar results can be achieved with lower doses of IL2, we have carried out a phase I/II trial of TIL transfer after classical lymphodepleting chemotherapy followed by an attenuated IL2 regimen. EXPERIMENTAL DESIGN: Twenty-five patients...... decrescendo regimen (ClinicalTrials.gov Identifier: NCT00937625). RESULTS: Classical IL2-related toxicities were observed but patients were manageable in a general oncology ward without the need for intervention from the intensive care unit. RECIST 1.0 evaluation displayed three complete responses and seven...... to treatment. CONCLUSIONS: TIL-ACT with a reduced IL2 decrescendo regimen results in long-lasting complete responses in patients with treatment-refractory melanoma. Larger randomized trials are needed to elucidate whether clinical efficacy is comparable with TIL-ACT followed by HD bolus IL2. Clin Cancer Res...

  3. Alterations in T cell-derived colony-stimulating factors associated with GVH-induced immune deficiency

    International Nuclear Information System (INIS)

    Injection of parental C57BL/10 spleen cells into unirradiated immune-competent (B10 x B10.BR)F1 hosts has been demonstrated to produce a graft-vs.-host-induced immune deficiency in T cell-mediated functions, including mitogen or alloantigen stimulated proliferation or cytotoxic T cell generation. The production of T cell-derived lymphokines affecting hematopoiesis was also altered during GVH. During the first two weeks of GVH, IL-3 and particularly GM-CSF were produced spontaneously; in subsequent weeks, the spontaneous production dropped to normal or subnormal levels. CSF content in concanavalin A-stimulated splenic supernatants was reduced at weeks 1-2, and declined to less than 5% of normal levels by 3-4 weeks of GVH. This decline in CSF content was correlated with a decrease in immune function as assessed by concanavalin A-stimulated IL-2 production and by generation of cytotoxic T lymphocytes. Concurrent with the recovery of immune function during GVH weeks 8-15, mitogen-stimulated production of CSF returned to normal levels. In addition to the decrease in CSF production identified in acute suppressive GVH, CSF content in concanavalin A-stimulated splenic supernatants was also decreased in chronic stimulatory GVH, generated in the strain combination (B6 x B6bm1)F1----(B6bm1 x B6bm12)F1. This decrease in CSF production correlated with a decrease in self-restricted T helper cell function. Finally, a decrease in both immune function and CSF production capacity was observed in the acute GVH following allogeneic (minor histocompatibility loci) bone marrow transplantation into irradiated hosts

  4. Functional Differences in Engineered Myocardium from Embryonic Stem Cell-Derived versus Neonatal Cardiomyocytes

    NARCIS (Netherlands)

    Feinberg, Adam W.; Ripplinger, Crystal M.; van der Meer, Peter; Sheehy, Sean P.; Domian, Ibrahim; Chien, Kenneth R.; Parker, Kevin Kit

    2013-01-01

    Stem cell-derived cardiomyocytes represent unique tools for cell-and tissue-based regenerative therapies, drug discovery and safety, and studies of fundamental heart-failure mechanisms. However, the degree to which stem cell-derived cardiomyocytes compare to mature cardiomyocytes is often debated. W

  5. Preparation of IL-2-loaded magnetic nanoparticle and its targeting accumulation in tumor tissues combining with external magnetic field%载IL-2磁性纳米粒的制备及其联合体外磁场在肿瘤组织中的靶向富集作用

    Institute of Scientific and Technical Information of China (English)

    沈爱蓉; 郭全义; 袁玫; 王桂琴; 卢世璧

    2013-01-01

    Objective: To prepare IL-2-loaded magnetic nanoparticle (IL-2-Fe3O4-PLGA) with Fe3O4 magnetic nanoparticle (Fe3O4-MNP) and poly lactide-co-glycolide (PLGA) , and to investigate its targeted accumulation function in tumor immunotherapy. Methods: The double emulsion method was employed for preparation of IL-2-Fe3O4-PLGA, its size and morphology were observed with a laser particle size analyzer and under a scanning electron microscope, respectively. In addition, the drug encapsulation efficiency and releasing characteristics of IL-2-Fe3O4-PLGA in vitro were measured with ELISA. The biological activity of the IL-2 released from the IL-2-Fe3O4-PLGA was evaluated by MTT assay. Sarcoma 180 cell-transplanted tumor mouse model was established to study the effect of IL-2-Fe3O4-PLGA in combination with external magnetic field on the growth of Sarcoma 180 tumors. The tumor, liver and kidney tissues were examined by Prussian blue staining to determine the accumulation and distribution of the IL-2-Fe3O4-PLGA in the tissue sections. Results: IL-2-Fe3O4-PLGA was constructed successfully, and was spherical with a mean diameter of (697 ±0. 51) nm as well as a drug encapsulation efficiency of (83. 76 ± 1. 24 ) % . In the drug release test in vitro, the mass concentration of released IL-2 was 100 ng/ml during the burst release phase and rose to 180 ng/ml at the end of 15 d. Moreover, the released IL-2 remained 85% -55% of its original activity during the releasing period. A strong IL-2-Fe3O4-PLGA positive reaction was observed in IL-2-Fe3O4-PLGA-treated mice combined with an external magnetic field, but only a weak reaction in mice injected with IL-2-Fe3O4-PLGA alone. Furthermore, in both groups, Prussian blue-stained liver showed occasionally light IL-2-Fe3O4-PLGA positive staining, while IL-2-Fe3O4-PLGA positive reaction was rarely detected in the kidneys. Conclusion: IL-2 loaded magnetic nanoparticle IL-2-Fe3O4-PLGA has controlled IL-2-release function and targeting accumulation

  6. IL-2和IL-10对Treg细胞体外扩增影响的差异研究%Effects of IL-2 and IL-10 on T regulatory cell proliferation in vitro

    Institute of Scientific and Technical Information of China (English)

    曾冬竹; 王自强; 郑峻松; 吴淼; 余佩武

    2007-01-01

    目的 分析细胞因子IL-2和IL-10对Treg细胞体外扩增的促进作用,比较扩增的Treg细胞和自然分选Treg细胞对CTL功能活性的影响.方法 采用不同浓度细胞因子IL-2和IL-10作为诱导剂,分析2种细胞因子体外诱导Treg细胞增殖的有效性及最适浓度,同时以2种细胞因子诱导扩增的Treg细胞和自然分选的Treg细胞分别作用于CTL,分析采用体外扩增Treg细胞抑制CTL活性的可能性.结果 100 ng/ml IL-2能够诱导Treg细胞增殖明显增加,经48、96 h和144 h诱导后,其增殖比例分别达到10%、18%和20%,而IL-10的使用浓度需达到150 ng/ml才能获得同样的扩增比例;与自然分选的Treg细胞对比,采用细胞因子体外扩增的Treg细胞具有同等抑制CTL功能活性的效力,组间无显著性差异(P>0.05).结论 采用添加IL-2培养体系是体外扩增Treg细胞的优选方法,其增殖效能强于IL-10;体外扩增Treg细胞与自然分选Treg细胞对CTL的功能抑制没有明显差异.

  7. Gamma c-signaling cytokines induce a regulatory T cell phenotype in malignant CD4+ T lymphocytes

    DEFF Research Database (Denmark)

    Kasprzycka, Monika; Zhang, Qian; Witkiewicz, Agnieszka;

    2008-01-01

    CD25 and TGF-beta, the expression of FOXP3 and, to a lesser degree, IL-10 was restricted to two CTCL cell lines that are dependent on exogenous IL-2. IL-2, IL-15, and IL-21, all of which signals through receptors containing the common gamma chain, induced expression of IL-10 in the IL-2-dependent...... that the T regulatory cell features are induced in CTCL T cells by common gamma chain signaling cytokines such as IL-2 and do not represent a fully predetermined, constitutive phenotype independent of the local environmental stimuli to which these malignant mature CD4(+) T cells become exposed....

  8. Combined stimulation of IL-2 and 4-1BB receptors augments the antitumor activity of E7 DNA vaccines by increasing Ag-specific CTL responses.

    Science.gov (United States)

    Kim, Ha; Kwon, Byungsuk; Sin, Jeong-Im

    2013-01-01

    Human papillomavirus (HPV) infection is a major cause of cervical cancer. Here, we investigate whether concurrent therapy using HPV E7 DNA vaccines (pE7) plus IL-2 vs. IL-15 cDNA and anti-4-1BB Abs might augment antitumor activity against established tumors. IL-2 cDNA was slightly better than IL-15 cDNA as a pE7 adjuvant. Co-delivery of pE7+IL-2 cDNA increased tumor cure rates from 7% to 27%, whereas co-delivery of pE7+IL-2 cDNA with anti-4-1BB Abs increased tumor cure rates from 27% to 67% and elicited long-term memory responses. This increased activity was concomitant with increased induction of Ag-specific CTL activity and IFN-γ responses, but not with Ag-specific IgG production. Moreover, the combined stimulation of IL-2 and 4-1BB receptors with rIL-2 and anti-4-1BB Abs resulted in enhanced production of IFN-γ from Ag-specific CD8+ T cells. However, this effect was abolished by treatment with anti-IL-2 Abs and 4-1BB-Fc, suggesting that the observed effect was IL-2- and anti-4-1BB Ab-specific. A similar result was also obtained for Ag-specific CTL activity. Thus, these studies demonstrate that combined stimulation through the IL-2 and 4-1BB receptors augments the Ag-specific CD8+ CTL responses induced by pE7, increasing tumor cure rates and long-term antitumor immune memory. These findings may have implications for the design of DNA-based therapeutic vaccines against cancer. PMID:24391824

  9. Combined stimulation of IL-2 and 4-1BB receptors augments the antitumor activity of E7 DNA vaccines by increasing Ag-specific CTL responses.

    Directory of Open Access Journals (Sweden)

    Ha Kim

    Full Text Available Human papillomavirus (HPV infection is a major cause of cervical cancer. Here, we investigate whether concurrent therapy using HPV E7 DNA vaccines (pE7 plus IL-2 vs. IL-15 cDNA and anti-4-1BB Abs might augment antitumor activity against established tumors. IL-2 cDNA was slightly better than IL-15 cDNA as a pE7 adjuvant. Co-delivery of pE7+IL-2 cDNA increased tumor cure rates from 7% to 27%, whereas co-delivery of pE7+IL-2 cDNA with anti-4-1BB Abs increased tumor cure rates from 27% to 67% and elicited long-term memory responses. This increased activity was concomitant with increased induction of Ag-specific CTL activity and IFN-γ responses, but not with Ag-specific IgG production. Moreover, the combined stimulation of IL-2 and 4-1BB receptors with rIL-2 and anti-4-1BB Abs resulted in enhanced production of IFN-γ from Ag-specific CD8+ T cells. However, this effect was abolished by treatment with anti-IL-2 Abs and 4-1BB-Fc, suggesting that the observed effect was IL-2- and anti-4-1BB Ab-specific. A similar result was also obtained for Ag-specific CTL activity. Thus, these studies demonstrate that combined stimulation through the IL-2 and 4-1BB receptors augments the Ag-specific CD8+ CTL responses induced by pE7, increasing tumor cure rates and long-term antitumor immune memory. These findings may have implications for the design of DNA-based therapeutic vaccines against cancer.

  10. Effect of Chinese Herbal Medicinal Ingredients on IL-2 mRNA Levels of T Lymphocytes in Mice Measured Using Semiquantification RT-PCR

    Institute of Scientific and Technical Information of China (English)

    CHU Yue-feng; YAN Xin-min; LI Xiang-rui; HU Yuan-liang

    2006-01-01

    In this study, the IL-2 mRNA levels of T lymphocytes in normal mice stimulated by nine Chinese herbal medicinal ingredients (CHMIs) were measured using semiquantification reverse transcription polymerase chain reaction. The results showed that astragalus polysaccharide (APS), epimedium polysaccharide (EPS), Chinese angelica polysaccharide (CAPS), propolis flavone (PF), and astrogalosides (AS) promoted IL-2 mRNA levels in T lymphocytes in vitro and in vivo to differing degrees, and the level of IL-2 mRNA induced by propolis polysaccharide (PPS) in vitro was higher than that induced by the control, which differed from that of PPS in vivo.

  11. Three agonist antibodies in combination with high-dose IL-2 eradicate orthotopic kidney cancer in mice

    Directory of Open Access Journals (Sweden)

    Sharkey Janelle

    2010-04-01

    Full Text Available Abstract Background Combination immunotherapies can be effective against subcutaneous tumors in mice but the effect against orthotopic malignant disease is less well characterized. In particular, a combination of three agonist antibodies, termed Tri-mAb, consisting of anti-DR5, anti-CD40 and anti-CD137 has previously been demonstrated to eradicate a large proportion of subcutaneous renal cell carcinoma (Renca tumors (75% long-term survival, but the effect against orthotopic disease is not known. Purpose To determine the relative response of orthotopic tumors, we inoculated Renca into the kidney followed by treatment with Tri-mAb. Results We found that orthotopic tumors responded much less to treatment (~13% survival, but a significant improvement in survival was achieved through the addition of IL-2 to the treatment regimen (55% survival. All three agonist antibodies and high dose IL-2, 100,000 IU for up to six doses, were required. CD8+ T cells were also required for optimal anti-tumor responses. Coadministration of IL-2 led to enhanced T cell activity as demonstrated by an increased frequency of IFN-gamma-producing T cells in tumor-draining lymph nodes, which may have contributed to the observed improvement of therapy against kidney tumors. Implications Responses of subcutaneous tumors to immunotherapy do not necessarily reflect how orthotopic tumors respond. The use of combination immunotherapy stimulating multiple facets of immunity and including cytokine support for T cells can induce effective anti-tumor responses against orthotopic and metastatic tumors.

  12. The influence of radiotherapy on IL-2 and IL-6 secretions of mucous membrane epithelial cells of wistar small intestine.

    Science.gov (United States)

    Liu, Bin; Li, Xiaoling; Ai, Fulu; Wang, Tianlu; Chen, Yun; Zhang, Hao

    2015-01-01

    The aim of the study was to investigate the influence of radiotherapy on IL-2 and IL-6 secretions of mucous epithelial cells of small intestine and the inhibition effect of deproteinized calf blood extractive (DCBE, also known as Actovegin in trade name) on apoptosis of mucous epithelial cells of small intestine. 50 wistars were randomly divided into 5 groups with 10 in each including normal group (NG), radiation group (RG), low-dose Actovegin group (L-AG), middle-dose Actovegin group (M-AG), and high-dose Actovegin (H-AG). High-energy X-ray linear accelerator was used for abdominal irradiation of RG, L-AG, M-AG, and H-AG at the exposure dose of 9.0 Gy to establish the wistar radiation damage model. Modeling wistars were injected with medicine for successive 4 days, and their small intestinal mucosas were extracted as pathological sections; then fully automated analyzer was employed to detect their IL-2 and IL-6 levels. Immunohistochemical analysis was carried out to explore the effect of Actovegin on apoptosis of mucous membrane epithelial cells of small intestine. The IL-2 and IL-6 levels of RG are significantly higher than other groups and differences are statistically significant (P 0.05). Compared with RG, the villus height, membrane thickness, crypt depth, and whole layer thickness significantly improved (P < 0.05). However, the expression levels of apoptosis-related protein bax of M-AG and H-AG are significantly lower than RG, and their bcl-2 levels are higher than RG with significant difference between them (P < 0.05). Actovegin is capable of effectively inhibiting the expression of apoptosis-related protein bax and facilitating the expression of anti-apoptosis protein bcl-2, having preferable remediation effect on mucous membrane epithelial cells of radioactive enteritis.

  13. The Experimental Study on Treating Transgenic HBV Mice with Recombined IL-2-PreS DNA Vaccine

    Institute of Scientific and Technical Information of China (English)

    李建远; 王海燕; 沈肖方; 王学波; 靳绍华; 刘芙君; 刘运祥

    2004-01-01

    The aim of this study is to investigate the feasibility and mechanism of hIL-2-preS DNA vaccine as prevention and therapeutic approach against Hepatitis B. Eukaryon expression vector involving hIL-2 and preS gene was constructed with recombinant technique and transferred into normal BALB/c mice and HBV transgenic mice (Tg-Mice) respectively. Tnen a series of detection were performed: detection of anti-preS2, HBs antibody and HBsAg in BALB/c mice and Tg-mice with ELISA, quantification of HBV DNA copies in HBV Tg-mice serum with real-time PCR, determination of hepatitis degree with immunopathological HE staining and detection of liver function. Anti-preS1 can be detected at 4th , 6th and 10th week in inoculated BALB/c mice. Injection with gene gun gained an advantage over muscular and subcutaneous injection since it acquired just 1/10 inoculation quantity (10μg/mouse). Highest expression of IgG2a at 4th week suggested Thl-mediated immune response, which facilitated HBV cleaning. Of all inoculated HBV Tg-mice, 80% of them showed anfi-preS2, HBs antibody positive and HBV DNA decreased, and 20% showed negative for HBsAg. HE staining to hepatic tissue showed obvious infiltration of inflammatory cells, swelling and granular degeneration of hepatocytes. In our study, IL-2-preS DNA vaccine which can provoke the humoral and cellular immune response and break the immune tolerance supports the designation and construction of new vaccine against HBV and specific immune remedy for HBV continuous infection.

  14. Metabolic and Signaling Functions of Cancer Cell-Derived Extracellular Vesicles.

    Science.gov (United States)

    Fonseca, P; Vardaki, I; Occhionero, A; Panaretakis, T

    2016-01-01

    Extracellular vesicles have gained tremendous attention in the recent years as a novel mechanism of cell to cell communication. There are several types of extracellular vesicles, including exosomes, microvesicles, exosome, like vesicles, apoptotic bodies that differ mainly in the mechanism of biogenesis and secretion. The most well studied type of extracellular vesicles are the exosomes which are endosome-derived vesicles with a diameter of 50-150nm and enriched in ESCRT proteins including Alix, TSG101, Hsp70, and tetraspanins. It is now well established that exosomes promote tumor growth, alter the tumor microenvironment, facilitate the dissemination of cancer cells in an organotropic manner, modulate immune responses, and mediate resistance to therapy. Exosomes have also been recently implicated in an emerging hallmark of cancer, the cancer cell metabolism. The metabolic state of the cell defines, to a certain extent, both the rate of secretion and the molecular content of tumor-derived exosomes. Furthermore, exosomes have been shown to possess intrinsic metabolic activity since they can synthesize ATP by glycolysis. It follows that exosomes carry a number of metabolic enzymes and metabolites, including lactate, PGE, LDH isoforms, pyruvate, and monocarboxylate transporters. Last but not the least, exosomes are implicated in fatty acid synthesis and cholesterol metabolism and are thought to be crucial for the transcellular metabolism procedure. Uptake of exosomes is thought to alter the intracellular metabolic state of the cell. In summary, we describe the state of the art on the role of metabolism in the secretion, uptake, and the biological effects of exosomes in the metabolism of recipient cells. PMID:27572129

  15. Nitric oxide regulates cell behavior on an interactive cell-derived extracellular matrix scaffold.

    Science.gov (United States)

    Xing, Qi; Zhang, Lijun; Redman, Travis; Qi, Shaohai; Zhao, Feng

    2015-12-01

    During tissue injury and wound healing process, there are dynamic reciprocal interactions among cells, extracellular matrix (ECM), and mediating molecules which are crucial for functional tissue repair. Nitric oxide (NO) is one of the key mediating molecules that can positively regulate various biological activities involved in wound healing. Various ECM components serve as binding sites for cells and mediating molecules, and the interactions further stimulate cellular activities. Human mesenchymal stem cells (hMSCs) can migrate to the wound site and contribute to tissue regeneration through differentiation and paracrine signaling. The objective of this work was to investigate the regulatory effect of NO on hMSCs in an interactive ECM-rich microenvironment. In order to mimic the in vivo stromal environment in wound site, a cell-derived ECM scaffold that was able to release NO within the range of in vivo wound fluid NO level was fabricated. Results showed that the micro-molar level of NO released from the ECM scaffold had an inhibitory effect on cellular activities of hMSCs. The NO impaired cell growth, altered cell morphology, disrupted the F-actin organization, also decreased the expression of focal adhesion related molecules integrin α5 and paxillin. These results may contribute to the elucidation of how NO acts on hMSCs in wound healing process.

  16. Construction of prokaryotic expression vector of melittin fused with a hIL-2 mutant%蜂毒肽与变构hIL-2融合基因原核表达载体的构建

    Institute of Scientific and Technical Information of China (English)

    郑旭; 鲁晓晴; 赵巍; 闫志勇; 钱冬萌; 丁守怡; 宋旭霞; 徐莉莉; 李鹏; 王斌

    2008-01-01

    目的 构建蜂毒肽与人变构白细胞介素-2原核表达载体,为进一步研究该融合基因而奠定基础.方法 以本室构建质粒pGEX-4T-2/Melittin-IL-2(88Arg)为模板,设计引物,用PCR定点突变的方法将IL-2部分的125Cys突变为Ala,PCR产物与pMD18-T连接,转化大肠杆菌,小提质粒,DNA测序后再将目的片段连接于pET-15b,最后酶切鉴定.结果 PCR产物542bp,构建质粒双酶切、DNA测序鉴定如预期.结论 成功构建蜂毒肽与人变构白细胞介素-2原核表达载体.

  17. Β-amyloid 1-42 oligomers impair function of human embryonic stem cell-derived forebrain cholinergic neurons.

    Directory of Open Access Journals (Sweden)

    Linn Wicklund

    Full Text Available Cognitive impairment in Alzheimer's disease (AD patients is associated with a decline in the levels of growth factors, impairment of axonal transport and marked degeneration of basal forebrain cholinergic neurons (BFCNs. Neurogenesis persists in the adult human brain, and the stimulation of regenerative processes in the CNS is an attractive prospect for neuroreplacement therapy in neurodegenerative diseases such as AD. Currently, it is still not clear how the pathophysiological environment in the AD brain affects stem cell biology. Previous studies investigating the effects of the β-amyloid (Aβ peptide on neurogenesis have been inconclusive, since both neurogenic and neurotoxic effects on progenitor cell populations have been reported. In this study, we treated pluripotent human embryonic stem (hES cells with nerve growth factor (NGF as well as with fibrillar and oligomeric Aβ1-40 and Aβ1-42 (nM-µM concentrations and thereafter studied the differentiation in vitro during 28-35 days. The process applied real time quantitative PCR, immunocytochemistry as well as functional studies of intracellular calcium signaling. Treatment with NGF promoted the differentiation into functionally mature BFCNs. In comparison to untreated cells, oligomeric Aβ1-40 increased the number of functional neurons, whereas oligomeric Aβ1-42 suppressed the number of functional neurons. Interestingly, oligomeric Aβ exposure did not influence the number of hES cell-derived neurons compared with untreated cells, while in contrast fibrillar Aβ1-40 and Aβ1-42 induced gliogenesis. These findings indicate that Aβ1-42 oligomers may impair the function of stem cell-derived neurons. We propose that it may be possible for future AD therapies to promote the maturation of functional stem cell-derived neurons by altering the brain microenvironment with trophic support and by targeting different aggregation forms of Aβ.

  18. 狼疮性肾炎患者血清IL-6、sIL-2R水平测定

    Institute of Scientific and Technical Information of China (English)

    胡波; 罗敏琪; 粱家隐; 叶少雄; 李朝霞

    2004-01-01

    To investigate the Changes of sIL - 2R, IL - 6 level in ,serum of patients with lupus nephritis and its clinical meaning. Methods The level of serum sIL - 2R, IL - 6 in 42 cases of lupus nephritis were detected by ELISA, and the data were compared with control group. Results Serum sIL- 2R ,IL-6 levels in lupus nephritis patients were significantly higher than control group and related to convalescent of the disease. Conclusion It suggested that immune dysfunction exist in patients with lupus nephritis higher level of sIL-2 ,IL-6 would associate with genesis and development of lupus nephritis, and it can be a new indicator of lupus nephritis.

  19. Soluble intercellular adhesion molecule-1 (sICAM-1) and soluble interleukin-2 receptors (sIL-2R) in scleroderma skin

    DEFF Research Database (Denmark)

    Søndergaard, Klaus; Deleuran, Mette; Heickendorff, Lene;

    1998-01-01

    In order to investigate whether soluble intercellular adhesion molecule-1 (sICAM-1) and soluble interleukin-2 receptors (sIL-2R) were present in scleroderma skin, and to compare their levels to concentrations measured in plasma and clinical parameters, we examined suction blister fluid and plasma...... from 13 patients with systemic sclerosis and 11 healthy volunteers. Suction blisters and biopsies were from the transition zone between normal skin and scleroderma, and uninvolved abdominal skin. The levels of sICAM-1 and sIL-2R were significantly increased in both plasma and suction blister fluid from...... systemic sclerosis patients compared with healthy volunteers. ICAM-1 was localized to vessels and perivascular mononuclear infiltrates by immunohistochemical methods. IL-2R was expressed by CD3-positive cells. The elevated levels of sICAM-1 and sIL-2R in suction blister fluid point towards activation...

  20. Study on Cytokines IL-2, IL-6, IL-10 in Patients of Chronic Allergic Rhinitis Treated with Acupuncture

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Objectives: To observe the plasmatic concentration of IL-6, IL-10 and IL-2 in the patient of chronic allergic rhinitis before and after acupuncture therapy. Methods: Cytokine levels were determined before and after treatment in 30 healthy volunteers (Group A) and 90 patients of chronic allergic rhinitis (Group B) with an increased plasma IL-10 level. Group B was then divided into 3 subgroups: 30 patients treated with real acupuncture (Group B1); 30 patients treated with sham acupuncture (Group B2); 30 non-treated patients (Group B3). Results: The allergic subjects of group B1, compared with controls, showed a significant reduction of IL-10 after a specific treatment with acupuncture (P<0.05). On the other hand, in those patients treated with sham acupuncture (B2) as well as in non-treated patients (B3), the IL-10 values remained high and unchanged. There was a statistically significant change in IL-2 values at 24 hours (P<0.05) after real acupuncture (Groups A, B1), however the values remained within normal ranges. The IL-6 do not change after therapy. Conclusion: The acupuncture treatment can reduce plasmatic level of IL-10 in chronic allergic rhinitis.

  1. Typing of HLA class II and class I antigens using PHA-activated, IL-2-propagated T lymphocytes.

    Science.gov (United States)

    Leshem, B; Cohen, I; Sherman, L; Brautbar, C; Kedar, E

    1988-06-28

    We describe here a simple procedure, by which HLA class II antigens can be accurately and reliably identified in those patients where there is minimal or absent expression of HLA-DR,DQw antigens on B cells, or when the total number of leukocytes recovered from the patients do not permit reliable typing. Ficoll-Hypaque-separated peripheral blood mononuclear leukocytes, fresh or cryopreserved, were activated by PHA and then propagated in IL-2-containing medium until enough cells for typing were obtained (usually 7-14 days). At this stage, the cultured cells were shown to be primarily T cells (greater than 90% CD3+). Since the activated T cells propagate in the presence of IL-2, even a small number (10(4] of fresh or cryopreserved patients' cells suffice for this protocol. To date we have been able to successfully HLA-DR,DQw type 34/34 bone marrow transplantation candidates and 12/12 long-term dialysis patients, who were untypable using fresh cells. HLA-DR,DQw antigens on activated T cells from normal individuals were identical to those found on their uncultured B cells. In addition, class I antigens that were undetectable on the uncultured cells of one patient could be identified on activated T cells. The HLA antigens identified on the patients' activated T cells were confirmed by phenotypic analysis of cells from family members. PMID:3260612

  2. CD8+ T Cell Fate and Function Influenced by Antigen-Specific Virus-Like Nanoparticles Co-Expressing Membrane Tethered IL-2.

    Directory of Open Access Journals (Sweden)

    Daniela Wojta-Stremayr

    Full Text Available A variety of adjuvants fostering humoral immunity are known as of today. However, there is a lack of adjuvants or adjuvant strategies, which directly target T cellular effector functions and memory. We here determined whether systemically toxic cytokines such as IL-2 can be restricted to the site of antigen presentation and used as 'natural adjuvants'. Therefore, we devised antigen-presenting virus-like nanoparticles (VNP co-expressing IL-2 attached to different membrane-anchors and assessed their potency to modulate CD8+ T cell responses in vitro and in vivo. Efficient targeting of IL-2 to lipid rafts and ultimately VNP was achieved by fusing IL-2 at its C-terminus to a minimal glycosylphosphatidylinositol (GPI-anchor acceptor sequence. To identify optimal membrane-anchor dimensions we inserted one (1Ig, two (2Ig or four (4Ig immunoglobulin(Ig-like domains of CD16b between IL-2 and the minimal GPI-anchor acceptor sequence of CD16b (GPI. We found that the 2IgGPI version was superior to all other evaluated IL-2 variants (IL-2v in terms of its i degree of targeting to lipid rafts and to the VNP surface, ii biological activity, iii co-stimulation of cognate T cells in the absence of bystander activation and iv potency to induce differentiation and acquisition of CD8+ T cell effector functions in vitro and in vivo. In contrast, the GPI version rather favored memory precursor cell formation. These results exemplify novel beneficial features of membrane-bound IL-2, which in addition to its mere T cell stimulatory capacity include the induction of differential effector and memory functions in CD8+ T lymphocytes.

  3. Relations of transcription expression of IL-2 with nuclear factor of activated T cells as well as changes of C-Fos and C-Jun after trauma

    Institute of Scientific and Technical Information of China (English)

    罗艳; 梁华平; 胡承香; 徐祥; 王正国

    2002-01-01

    Objective: To observe the relations among expression of interleukin-2 (IL-2) in spleen lymphocytes, DNA binding activity of nuclear factor of activated T cells (NFAT) and expression of the partly family members C-Fos, C-Jun after trauma. Methods: A murine closed trauma model was used, animals were sacrificed 6, 12 hours and 1, 4, 7, 10, 14 days, respectively after injury. Spleen lymphocytes were isolated from injured mice and stimulated with concanavalin-A. The culture supernatants were harvested and assayed for IL-2 activity. Total RNA was extracted from spleen lymphocytes and assayed for IL-2 mRNA. Nuclear protein was extracted, and the DNA binding activity of NFAT was measured using an electrophoretic mobility shift assay (EMSA), the expressions of C-Fos, C-Jun protein determined by Western blot analysis. Results: The expressions of IL-2 activity and IL-2 mRNA in spleen lymphocytes were decreased in injured mice compared with those in control mice, and the most obvious decrease appeared on the 4th day after injury. The DNA binding activity of NFAT decreased gradually and reached the minimum that was only 41% of the control on the 4th day after injury, which was closely associated with the decline of IL-2 activity and IL-2 mRNA. An decrease in the expression of C-Fos on the 1st and 4th day after injury, trauma had no significant effect on the C-Jun expression.Conclusions: These results suggest that the inhibition of IL-2 expression is partly due to the impairment in the activation of NFAT in injured mice; and the decline in the DNA binding activity of NFAT is partly due to trauma block in the C-Fos expression.

  4. Detection of T lymphocyte subsets and mIL-2R on surface of PBMC in patients with hepatitis B

    Institute of Scientific and Technical Information of China (English)

    Ke-Xia Wang; Jiang-Long Peng; Xue-Feng Wang; Ye Tian; Jian Wang; Chao-Pin Li

    2003-01-01

    AIM: To study the levels of T lymphocyte subsets andmembrane interleukin-2 receptor (mIL-2R) on surface ofperipheral blood mononuclear cells (PBMCs) of patients withhepatitis B and its role in the pathogenesis of hepatitis B.METHODS: The levels of T lymphocyte subsets and mIL-2R in PBMC before and after being stimulated with PHAwere detected by biotin-streptavidin (BSA) technique in 196cases of hepatitis B.RESULTS: In patients with hepatitis B, the levels of CD3+,CD4+ cells, and the ratio of CD4+ cells/CD8+ cells were lower,but the level of CD8+ cells was higher than those in normalcontrols (42.20±6.01 vs65.96±6.54, 38.17±5.93 vs41.73±6.40,0.91±0.28 vs 1.44±0.31, 39.86±6.36 vs30.02±-4.54, P<0.01).The total expression level of mIL-2R in PBMC before andafter being stimulated with PHA was also lower than thosein normal controls (3.47±1.55 vs4.52±1.49, 34.03±2.94 vs37.95±3.00, P<0.01). In all the patients with hepatitis B, thelevels of T lymphocyte subsets and mIL-2R in PBMC withHBV-DNA (+) were lower than those with HBV-DNA (-),which were significantly different (39.57±7.11 vs44.36±5.43,34.36±7.16 vs 40.75±5.87, 37.82±6.54 vs 41.72±6.21,0.88±0.33 vs0.99±0.27, 2.82±1.62 vs3.85±1.47, 31.56±3.00vs35.84±2.83, P<0.01). In addition, the levels of CD3+, CD4+,CD8+ cells, the ratio of CD4+ cells/CD8+ cells and mIL-2R amongdifferent courses of hepatitis B were all significantly different(F=3 723.18, P<0.01. F=130.43, P<0.01. F=54.01, P<0.01.F=2.99, P<0.05. F=7.16, P<0.01).CONCLUSION: Both cellular and humoral immune functionsare obviously in disorder in patients with hepatitis B, whichmight be closely associated with the chronicity in patients.

  5. IL2RA/CD25 Gene Polymorphisms: Uneven Association with Multiple Sclerosis (MS) and Type 1 Diabetes (T1D)

    Science.gov (United States)

    Alcina, Antonio; Fedetz, María; Ndagire, Dorothy; Fernández, Oscar; Leyva, Laura; Guerrero, Miguel; Abad-Grau, María M.; Arnal, Carmen; Delgado, Concepción; Lucas, Miguel; Izquierdo, Guillermo; Matesanz, Fuencisla

    2009-01-01

    Background IL-2 receptor (IL2R) alpha is the specific component of the high affinity IL2R system involved in the immune response and in the control of autoimmunity. Methods and Results Here we perform a replication and fine mapping of the IL2RA gene region analyzing 3 SNPs previously associated with multiple sclerosis (MS) and 5 SNPs associated with type 1 diabetes (T1D) in a collection of 798 MS patients and 927 matched Caucasian controls from the south of Spain. We observed association with MS in 6 of 8 SNPs. The rs1570538, at the 3′- UTR extreme of the gene, previously reported to have a weak association with MS, is replicated here (P = 0.032). The most associated T1D SNP (rs41295061) was not associated with MS in the present study. However, the rs35285258, belonging to another independent group of SNPs associated with T1D, showed the maximal association in this study but different risk allele. We replicated the association of only one (rs2104286) of the two IL2RA SNPs identified in the recently performed genome-wide association study of MS. Conclusions These findings confirm and extend the association of this gene with MS and reveal a genetic heterogeneity of the associated polymorphisms and risk alleles between MS and T1D suggesting different immunopathological roles of IL2RA in these two diseases. PMID:19125193

  6. Correlation between gene polymorphisms of IL-2, IL-6 and TNF-α in workers exposed to rare earth dust and their lung ventilation function

    International Nuclear Information System (INIS)

    Objective: To study the relationship between lung ventilation function of workers exposed to rare earth dust and their IL-2, IL-6 and TNF-α gene polymorphism. Methods: TNF-α gene polymorphism were identified by RFLP-PCR, IL-2 and IL-6 gene polymorphisms were identified by PCR-CTPP analysis. Lung ventilation function was deteced by instrument of ventilation function. Results: Compared with controls, there was no statistic significance in frequencies distribution of TNF-α gene polymorphism (χ2=4.03, P>0.05), IL-2 gene polymorphism (χ2=2.21, P>0.05) and IL-6 gene polymorphism (χ2=1.05, P>0.05). Compared with IL-2 gene wild type, IL-2 homozygote type increased the risk of lung ventilation dysfunction by 4.29 folds (95% CI 1.09 ∼ 16.9). Conclusions: Compared with controls,incidence of ventilation function of workers exposed to rare earth dust is in ascending trend. IL-2 (G/G) gene type induces more serious inflammation reaction than the others. (authors)

  7. Inducing P-selectin ligand formation in CD8 T cells: IL-2 and IL-12 are active in vitro but not required in vivo.

    Science.gov (United States)

    Carlow, Douglas A; Williams, Michael J; Ziltener, Hermann J

    2005-04-01

    In vitro studies have demonstrated that IL-2 and IL-12 can support formation of P-selectin ligands (P-SelL) in activated T cells, ligands that are variably required for efficient lymphocyte recruitment to sites of inflammation. To ascertain whether these cytokines were required for P-SelL formation in vivo, TCR transgenic CD8 T cells specific for male Ag (HY) were transferred into male mice under conditions in which either IL-2 and/or IL-15 or IL-12Rp40 were absent. P-SelL formation at day 2 was unperturbed in HY-TCR IL-2(null) CD8 T cells responding in doubly deficient IL-2(null)IL-12(null) or IL-2(null)IL-15(null) male recipients. HY-specific CD8 T cell proliferative responses detected in both spleen and peritoneum occurred vigorously, but only splenic CD8 T cells up-regulated P-SelL, demonstrating that in vivo induction of P-SelL is an active, nonprogrammed event following T cell activation and that despite the efficacy of IL-2 and IL-12 in supporting P-SelL formation in vitro, these cytokines appear to be dispensable for this purpose in vivo.

  8. IL-4 function can be transferred to the IL-2 receptor by tyrosine containing sequences found in the IL-4 receptor alpha chain.

    Science.gov (United States)

    Wang, H Y; Paul, W E; Keegan, A D

    1996-02-01

    IL-4 binds to a cell surface receptor complex that consists of the IL-4 binding protein (IL-4R alpha) and the gamma chain of the IL-2 receptor complex (gamma c). The receptors for IL-4 and IL-2 have several features in common; both use the gamma c as a receptor component, and both activate the Janus kinases JAK-1 and JAK-3. In spite of these similarities, IL-4 evokes specific responses, including the tyrosine phosphorylation of 4PS/IRS-2 and the induction of CD23. To determine whether sequences within the cytoplasmic domain of the IL-4R alpha specify these IL-4-specific responses, we transplanted the insulin IL-4 receptor motif (I4R motif) of the huIL-4R alpha to the cytoplasmic domain of a truncated IL-2R beta. In addition, we transplanted a region that contains peptide sequences shown to block Stat6 binding to DNA. We analyzed the ability of cells expressing these IL-2R-IL-4R chimeric constructs to respond to IL-2. We found that IL-4 function could be transplanted to the IL-2 receptor by these regions and that proliferative and differentiative functions can be induced by different receptor sequences.

  9. Defined MicroRNAs Induce Aspects of Maturation in Mouse and Human Embryonic-Stem-Cell-Derived Cardiomyocytes

    Directory of Open Access Journals (Sweden)

    Desy S. Lee

    2015-09-01

    Full Text Available Pluripotent-cell-derived cardiomyocytes have great potential for use in research and medicine, but limitations in their maturity currently constrain their usefulness. Here, we report a method for improving features of maturation in murine and human embryonic-stem-cell-derived cardiomyocytes (m/hESC-CMs. We found that coculturing m/hESC-CMs with endothelial cells improves their maturity and upregulates several microRNAs. Delivering four of these microRNAs, miR-125b-5p, miR-199a-5p, miR-221, and miR-222 (miR-combo, to m/hESC-CMs resulted in improved sarcomere alignment and calcium handling, a more negative resting membrane potential, and increased expression of cardiomyocyte maturation markers. Although this could not fully phenocopy all adult cardiomyocyte characteristics, these effects persisted for two months following delivery of miR-combo. A luciferase assay demonstrated that all four miRNAs target ErbB4, and siRNA knockdown of ErbB4 partially recapitulated the effects of miR-combo. In summary, a combination of miRNAs induced via endothelial coculture improved ESC-CM maturity, in part through suppression of ErbB4 signaling.

  10. IL-2 immunotherapy reveals potential for innate beta cell regeneration in the non-obese diabetic mouse model of autoimmune diabetes.

    Directory of Open Access Journals (Sweden)

    Yaiza Diaz-de-Durana

    Full Text Available Type-1 diabetes (T1D is an autoimmune disease targeting insulin-producing beta cells, resulting in dependence on exogenous insulin. To date, significant efforts have been invested to develop immune-modulatory therapies for T1D treatment. Previously, IL-2 immunotherapy was demonstrated to prevent and reverse T1D at onset in the non-obese diabetic (NOD mouse model, revealing potential as a therapy in early disease stage in humans. In the NOD model, IL-2 deficiency contributes to a loss of regulatory T cell function. This deficiency can be augmented with IL-2 or antibody bound to IL-2 (Ab/IL-2 therapy, resulting in regulatory T cell expansion and potentiation. However, an understanding of the mechanism by which reconstituted regulatory T cell function allows for reversal of diabetes after onset is not clearly understood. Here, we describe that Ab/IL-2 immunotherapy treatment, given at the time of diabetes onset in NOD mice, not only correlated with reversal of diabetes and expansion of Treg cells, but also demonstrated the ability to significantly increase beta cell proliferation. Proliferation appeared specific to Ab/IL-2 immunotherapy, as anti-CD3 therapy did not have a similar effect. Furthermore, to assess the effect of Ab/IL-2 immunotherapy well after the development of diabetes, we tested the effect of delaying treatment for 4 weeks after diabetes onset, when beta cells were virtually absent. At this late stage after diabetes onset, Ab/IL-2 treatment was not sufficient to reverse hyperglycemia. However, it did promote survival in the absence of exogenous insulin. Proliferation of beta cells could not account for this improvement as few beta cells remained. Rather, abnormal insulin and glucagon dual-expressing cells were the only insulin-expressing cells observed in islets from mice with established disease. Thus, these data suggest that in diabetic NOD mice, beta cells have an innate capacity for regeneration both early and late in disease

  11. Higher susceptibility of NOD/LtSz-scid Il2rg-/- NSG mice to xenotransplanted lung cancer cell lines

    Directory of Open Access Journals (Sweden)

    Kanaji N

    2014-10-01

    Full Text Available Nobuhiro Kanaji,1 Akira Tadokoro,1 Kentaro Susaki,1 Saki Yokokura,1 Kiyomi Ohmichi,2 Reiji Haba,2 Naoki Watanabe,1 Shuji Bandoh,1 Tomoya Ishii,1 Hiroaki Dobashi,1 Takuya Matsunaga11Department of Internal Medicine, Division of Hematology, Rheumatology and Respiratory Medicine, Faculty of Medicine, Kagawa University, Kagawa, Japan; 2Department of Diagnostic Pathology, Faculty of Medicine, Kagawa University, Kagawa, JapanPurpose: No lung cancer xenograft model using non-obese diabetic (NOD-scid Il2rg-/- mice has been reported. The purpose of this study is to select a suitable mouse strain as a xenogenic host for testing tumorigenicity of lung cancer.Materials and methods: We directly compared the susceptibility of four immunodeficient mouse strains, c-nu, C.B-17 scid, NOD-scid, and NOD/LtSz-scid Il2rg-/- (NSG mice, for tumor formation from xenotransplanted lung cancer cell lines. Various numbers (101–105 cells/head of two lung cancer cell lines, A549 and EBC1, were subcutaneously inoculated and tumor sizes were measured every week up to 12 weeks.Results: When 104 EBC1 cells were inoculated, no tumor formation was observed in BALB/c-nu or C.B-17 scid mice. Tumors developed in two of the five NOD-scid mice (40% and in all the five NSG mice (100%. When 103 EBC1 cells were injected, no tumors developed in any strain other than NSG mice, while tumorigenesis was achieved in all the five NSG mice (100%, P=0.0079 within 9 weeks. NSG mice similarly showed higher susceptibility to xenotransplantation of A549 cells. Tumor formation was observed only in NSG mice after inoculation of 103 or fewer A549 cells (40% vs 0% in 15 NSG mice compared with others, respectively, P=0.0169. We confirmed that the engrafted tumors originated from inoculated human lung cancer cells by immunohistochemical staining with human cytokeratin and vimentin.Conclusion: NSG mice may be the most suitable strain for testing tumorigenicity of lung cancer, especially if only a few cells

  12. The multiple sclerosis susceptibility genes TAGAP and IL2RA are regulated by vitamin D in CD4+ T cells.

    Science.gov (United States)

    Berge, T; Leikfoss, I S; Brorson, I S; Bos, S D; Page, C M; Gustavsen, M W; Bjølgerud, A; Holmøy, T; Celius, E G; Damoiseaux, J; Smolders, J; Harbo, H F; Spurkland, A

    2016-03-01

    Multiple sclerosis (MS) is an inflammatory, demyelinating disorder of the central nervous system that develops in genetically susceptible individuals. The majority of the MS-associated gene variants are located in genetic regions with importance for T-cell differentiation. Vitamin D is a potent immunomodulator, and vitamin D deficiency has been suggested to be associated with increased MS disease susceptibility and activity. In CD4+ T cells, we have analyzed in vitro vitamin D responsiveness of genes that contain an MS-associated single-nucleotide polymorphism (SNP) and with one or more vitamin D response elements in their regulatory regions. We identify IL2RA and TAGAP as novel vitamin D target genes. The vitamin D response is observed in samples from both MS patients and controls, and is not dependent on the genotype of MS-associated SNPs in the respective genes. PMID:26765264

  13. Fine mapping and trans-ethnic genotyping establish IL2/IL21 genetic association with lupus and localize this genetic effect to IL21

    Science.gov (United States)

    Hughes, Travis; Kim-Howard, Xana; Kelly, Jennifer A.; Kaufman, Kenneth M.; Langefeld, Carl D.; Ziegler, Julie; Sanchez, Elena; Kimberly, Robert P.; Edberg, Jeffrey C.; Ramsey-Goldman, Rosalind; Petri, Michelle; Reveille, John D.; Martin, Javier; Brown, Elizabeth E.; Vilá, Luis M.; Alarcón, Graciela S.; James, Judith A.; Gilkeson, Gary S.; Moser, Kathy L.; Gaffney, Patrick M.; Merrill, Joan T.; Vyse, Timothy J.; Alarcón-Riquelme, Marta E.; Nath, Swapan K.; Harley, John B.; Sawalha, Amr H.

    2011-01-01

    Objective Genetic association of the IL2/IL21 region at 4q27 has been previously reported in lupus and a number of autoimmune and inflammatory diseases. Herein, using a very large cohort of lupus patients and controls, we localize this genetic effect to the IL21 gene. Methods We genotyped 45 tag SNPs across the IL2/IL21 locus in two large independent lupus sample sets. We studied a European-derived set consisting of 4,248 lupus patients and 3,818 healthy controls, and an African-American set of 1,569 patients and 1,893 healthy controls. Imputation in 3,004 WTCCC additional control individuals was also performed. Genetic association between the genotyped markers was determined, and pair-wise conditional analysis was performed to localize the independent genetic effect in the IL2/IL21 locus in lupus. Results We established and confirmed the genetic association between IL2/IL21 and lupus. Using conditional analysis and trans-ethnic mapping, we localized the genetic effect in this locus to two SNPs in high linkage disequilibrium; rs907715 located within IL21 (OR=1.16 (1.10–1.22), P= 2.17 ×10−8), and rs6835457 located in the 3’-UTR flanking region of IL21 (OR= 1.11 (1.05–1.17), P= 9.35×10−5). Conclusion We have established the genetic association between lupus and IL2/IL21 with a genome-wide level of significance. Further, we localized this genetic association within the IL2/IL21 linkage disequilibrium block to IL21. If other autoimmune IL2/IL21 genetic associations are similarly localized, then the IL21 risk alleles would be predicted to operate in a fundamental mechanism that influences the course of a number of autoimmune disease processes. PMID:21425124

  14. EFFICIENT ACTIVATION OF ANTITUMOR IMMUNITY BY IL-6 GENE-MODIFIED LEUKEMIA VACCINE IN COMBINATION WITH LOW DOSE CYCLOPHOSPHAMIDE AND LOW DOSE IL-2

    Institute of Scientific and Technical Information of China (English)

    Cao Xuetao; Ge Lingfu; Ju Dianwen; Tao Qun; Yu Yizhi

    1998-01-01

    Objective:To investigate the antitumor effect of the IL-6 gene-modified erythroleukemia cells combined with low dose cyclophosphamide (Cy) and low dose IL-2.Methods: Mice inoculated with FBL-3-IL-6 in combination with low dose IL-2 and low dose cyclophosphamide (Cy). Results: Mice received combined therapy of FBL-3-IL-6, IL-2 and Cy developed tumors most slowly and survived much longer when compared with mice in control groups, with 5 out of 8leukemia-bearing mice being tumor free 100 days after the combined treatment. To further explain the mechanism of the antitumor effects by the combined therapy. It was found that combined therapy with low dose Cy, low dose IL-2 and FBL-3-IL-6 achieved maximal cytotoxic effects of nature killer cells and specific cytotoxic T lymphocytes, increased production IL-2, TNF and GM-CSF from spleen lymphocytes in tumor-bearing mice. Vaccination with the FBL3-IL-6 also enhanced the cytotoxic activity of the peritoneal macrophages. The results demonstrated that administration of low dose Cy and low dose IL-2 in combination with IL-6 genemodified leukemia vaccine could elicit potent antileukemia effects, and the mechanisms involved in the antitumor process may include the induction of specific and nonspecific antitumor immunity, reversal of T suppressor cells that mediated local immuno-suppression in tumor bearing mice. Conclusion: The combined therapy with cytokine gene-modified tumor vaccine, low dose of Cy and IL-2 might be a promising approach for the treatment of leukemia.

  15. Immunological Properties of Recombinant Mycobacterium bovis Bacillus Calmette-Guérin Strain Expressing Fusion Protein IL-2-ESAT-6

    Institute of Scientific and Technical Information of China (English)

    Xiong-Lin FAN; Ting-He YU; Qian GAO; Wei YAO

    2006-01-01

    The live vaccine Mycobacterium bovis bacillus Calmette-Guérin (BCG) provides variable efficacy against adult pulmonary tuberculosis (TB). Recombinant BCG, expressing either immunodominant antigens or Thl cytokines, is a promising trategy for developing a new TB vaccine. However, not much is known about whether the introduction of cytokine and specific antigen genes concurrently into the BCG strain could improve the munogenicity of BCG. In this study, a recombinant BCG strain (rBCG) expressing the fusion protein human interleukin (IL)-2 and ESAT-6 (early secreted antigenic target-6 kDa) antigen of ycobacterium tuberculosis was constructed. Six weeks after BALB/c mice (H-2d) were immunized with 106 colony forming units (CFUs) BCG or rBCG, splenocyte proliferation was determined with MTT[3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide] assay, IL-4 and interferon (IFN)-γ produced by plenocytes were tested by enzyme linked immunosorbent assay (ELISA,) and the cytotoxicity of splenocytes from immunized mice to P815 cells (H-2d) expressing ESAT-6 protein was measured using CytoTox 96 Non-Radioactive Cytotoxicity Assay. Compared with native BCG-vaccinated mice, rBCG induced stronger Th1 responses that were confirmed by high lymphoproliferative responses and IFN-γ production to culture filtrate protein (CFP) or ESAT-6 protein. Moreover, rBCG induced significant enhanced CTL responses against P815-ESAT-6 cells. Results from rBCG-immunized mice demonstrated that introducing the il-2 and esat-6 genes into BCG could enhance Th1 type immune responses to ESAT-6. Further investigation is needed by introducing other Th1 cytokines and antigens into BCG to optimize the protective efficacy against TB.

  16. Knockout of endothelial cell-derived endothelin-1 attenuates skin fibrosis but accelerates cutaneous wound healing.

    Directory of Open Access Journals (Sweden)

    Katsunari Makino

    Full Text Available Endothelin (ET-1 is known for the most potent vasoconstrictive peptide that is released mainly from endothelial cells. Several studies have reported ET-1 signaling is involved in the process of wound healing or fibrosis as well as vasodilation. However, little is known about the role of ET-1 in these processes. To clarify its mechanism, we compared skin fibrogenesis and wound repair between vascular endothelial cell-specific ET-1 knockout mice and their wild-type littermates. Bleomycin-injected fibrotic skin of the knockout mice showed significantly decreased skin thickness and collagen content compared to that of wild-type mice, indicating that bleomycin-induced skin fibrosis is attenuated in the knockout mice. The mRNA levels of transforming growth factor (TGF-β were decreased in the bleomycin-treated skin of ET-1 knockout mice. On the other hand, skin wound healing was accelerated in ET-1 knockout mice, which was indicated by earlier granulation tissue reduction and re-epithelialization in these mice. The mRNA levels of TGF-β, tumor necrosis factor (TNF-α and connective tissue growth factor (CTGF were reduced in the wound of ET-1 knockout mice. In endothelial ET-1 knockout mouse, the expression of TNF-α, CTGF and TGF-β was down-regulated. Bosentan, an antagonist of dual ET receptors, is known to attenuate skin fibrosis and accelerate wound healing in systemic sclerosis, and such contradictory effect may be mediated by above molecules. The endothelial cell-derived ET-1 is the potent therapeutic target in fibrosis or wound healing, and investigations of the overall regulatory mechanisms of these pathological conditions by ET-1 may lead to a new therapeutic approach.

  17. Effector cells derived from naive T cells used in tumor immunotherapy of mice bearing B16 melanoma

    Institute of Scientific and Technical Information of China (English)

    Wen Ming; Xu Weili; Ren Lili; Gao Fei; Cui Naipeng; Wen Junye; Li Xinjiang

    2014-01-01

    Background Adoptive cell transfer (ACT) immunotherapy has been used clinically for years to treat malignancies.Improving the killing efficiency of effector cells,such as tumor-specific cytotoxic T lymphocytes (CTLs),is an important component for enhancing the clinical response of cancer immunotherapy.Hence,we explored a novel method for preparing cancer-specific CTLs using naive T lymphocytes.Methods C57BL/6 mice bearing B16 melanoma tumors were pretreated with cyclophosphamide (CTX) by peritoneal injection.The immunosuppressive influence of CTX on tumor regression and the tumor microenvironment was assessed.Naive T cells and T cell pools were isolated via negative selection using immunomagnetic beads.The proliferative potential and cytokine production of different T cell subpopulations were evaluated in vitro.Tumor-specific CTLs derived from naive T cells (naive CD4+ T cells:naive CD8+ T cells=2:1) and pooled T cells were generated in vitro,respectively.B16 melanoma-bearing C57BL/6 mice were pretreated with CTX,followed by ACT immunotherapy using dendritic cell-induced CTLs.The homing abilities of the effector cells and interleukin-2 (IL-2),interferon-y,granzyme B,and perforin mRNA levels in tumor tissues were evaluated,and the change in tumor volume was measured.Results Mice receiving CTX peritoneal pretreatment injections did not display tumor regression compared with control mice.However,a significant downregulation of splenic Tregs and tumor growth factor-β1 (TGF-β1) and interleukin-10 (IL-10) serum levels was observed (P <0.05).Naive T cells showed a stronger proliferative capacity and elevated cytokine production than did pooled T cells (P <0.05).In addition,effector cells generated from naive T cells displayed more potent antitumor activity in vivo than those derived from pooled T cells (P <0.05).Conclusion Effector cells derived from the naive T cells possess a stronger proliferative potential,homing capacity,and enhanced cytokine production

  18. Mesenchymal stem cells derived from human exfoliated deciduous teeth (SHEDs induce immune modulatory profile in monocyte-derived dendritic cells.

    Directory of Open Access Journals (Sweden)

    Fernando de Sá Silva

    Full Text Available BACKGROUND: Mesenchymal stem cells have prominent immune modulatory properties, which may have clinical applications; however their major source, bone marrow, is of limited availability. On the other hand, mesenchymal stem cells derived from human exfoliated deciduous teeth (SHEDs are readily accessible, but their immune regulatory properties have not been completely investigated. This study was designed, therefore, to evaluate the SHEDs influence on DCs differentiation, maturation, ability to activate T cells and to expand CD4(+Foxp3(+ T cells. METHODOLOGY/PRINCIPAL FINDINGS: The experiments were based in cellular co-culture during differentiation and maturation of monocyte derived-DCs (moDCs, with, or not, presence of SHEDs. After co-culture with SHEDs, (moDCs presented lower expression of BDCA-1 and CD11c, in comparison to DC cultivated without SHEDs. CD40, CD80, CD83 and CD86 levels were also decreased in mature DCs (mDCs after co-cultivation with SHEDs. To assess the ability of SHEDs-exposed moDCs to modulate T cell responses, the former were separated from SHEDs, and co-cultured with peripheral blood lymphocytes. After 5 days, the proliferation of CD4(+ and CD8(+ T cells was evaluated and found to be lower than that induced by moDCs cultivated without SHEDs. In addition, an increase in the proportion of CD4(+Foxp3(+IL-10(+ T cells was observed among cells stimulated by mature moDCs that were previously cultivated with SHEDs. Soluble factors released during co-cultures also showed a reduction in the pro-inflammatory cytokines (IL-2, TNF-α and IFN-γ, and an increase in the anti-inflammatory molecule IL-10. CONCLUSION/SIGNIFICANCE: This study shows that SHEDs induce an immune regulatory phenotype in moDCs cells, evidenced by changes in maturation and differentiation rates, inhibition of lymphocyte stimulation and ability to expand CD4(+Foxp3(+ T cells. Further characterization and validation of this phenomenon could support the use of SHEDs

  19. An Abbreviated Protocol for In Vitro Generation of Functional Human Embryonic Stem Cell-Derived Beta-Like Cells

    DEFF Research Database (Denmark)

    Massumi, Mohammad; Pourasgari, Farzaneh; Nalla, Amarnadh;

    2016-01-01

    developed an abbreviated five-stage protocol (25-30 days) to generate human Embryonic Stem Cell-Derived Beta-like Cells (ES-DBCs). We showed that Geltrex, as an extracellular matrix, could support the generation of ES-DBCs more efficiently than that of the previously described culture systems......The ability to yield glucose-responsive pancreatic beta-cells from human pluripotent stem cells in vitro will facilitate the development of the cell replacement therapies for the treatment of Type 1 Diabetes. Here, through the sequential in vitro targeting of selected signaling pathways, we have...... positive cells, 1% insulin and glucagon positive cells and 30% insulin and NKX6.1 co-expressing cells. Functionally, ES-DBCs were responsive to high glucose in static incubation and perifusion studies, and could secrete insulin in response to successive glucose stimulations. Mitochondrial metabolic flux...

  20. Prevention of hepatocellular carcinoma in mice by IL-2 and B7-1genes co-transfected liver cancer cell vaccines

    Institute of Scientific and Technical Information of China (English)

    Ning-Ling Ge; Sheng-Long Ye; Ning Zheng; Rui-Xia Sun; Yin-Kun Liu; Zhao-You Tang

    2003-01-01

    AIM: To study the immunoprotective effect of liver cancer vaccine with co-transfected IL-2 and B7-1 genes on hepatocarcinogenesis in mice.METHODS: The murine liver cancer cell line Hepal-6 was transfected with IL-2 and/or B7-1 gene via recombinant adenoviral vectors and the liver cancer vaccines were prepared. C57BL/6 mice were immunized with these vaccines and challenged with the parental Hepal-6 cells afterwards.The immunoprotection was investigated and the reactive T cell line was assayed.RESULTS: The immunoprotection of the tumor vaccine was demonstrated. The effect of IL-2 and B7-1 genes cotransfected Hepal-6 liver cancer vaccine (Hep6-IL2/B7vaccine) on the onset of tumor formation was the strongest.When attacked with wild Hepal-6 cells, the median survival period of the mice immunized with Hep6-IL2/B7 vaccine was the longest (68 days, χ2=7.70-11.69, P<0.05) and the implanted tumor was the smallest (z =3.20-44.10, P<0.05).The effect of single IL-2 or B7-1 gene-transfected vaccine was next to the IL2/B7 gene co-transfected group, and the mean survival periods were 59 and 54 days, respectively.The mean survival periods of wild or enhanced green fluorescence protein gene modified vaccine immunized group were 51 and 48 days, respectively. The mice in control group all died within 38 days and the implanted tumor was the largest (z=3.20-40.21, P<0.05). The cellular immunofunction test and cytotoxicity study showed that the natural killer (NK) cell, lymphokine activated killer (LAK) cell and cytotoxic T lymphocyte (CTL) activities were significantly increased in mice immunized with the Hep6-IL2/B7 vaccine, (29.5±2.5%,65.0±2.9%, 83.1±1.5% respectively, compared with other groups, P<0.05).CONCLUSION: The Hep6-IL2/B7 liver cancer vaccines can induce the mice to produce activated and specific CTL against the parental tumor cells, and demonstrate stronger effect on the hepatocarcinogenesis than single gene modified or the regular tumor vaccine. Therefore, the

  1. 胃癌患者血清中 IL-2,IL-4和 IL-6的动态分析及临床意义

    Institute of Scientific and Technical Information of China (English)

    孙景春; 李锐

    2013-01-01

    目的:探讨胃癌患者血清中 IL-2,IL-4和 IL-6的动态变化及临床意义;方法:应用 ELISA 的方法检测手术前后胃癌患者血清中 IL-2, IL-4和 IL-6的水平,并与良性肿瘤患者和健康体检者(对照组)进行比较.结果:胃癌患者在手术前,IL-4和 IL-6的水平高于良性肿瘤组和对照组(P<0.05),IL-2的水平低于良性肿瘤组和对照组(P<0.05);而在手术后 IL-4和 IL-6的水平明显降低,IL-2的水平呈现升高趋势,与对照组水平接近.结论:胃癌病人存在免疫功能的紊乱,IL-2,IL-4和 IL-6的水平变化参与了胃癌的发生、发展过程,对胃癌患者的诊断和预后判断具有重要的指导意义.

  2. Involvement of multiple myeloma cell-derived exosomes in osteoclast differentiation.

    Science.gov (United States)

    Raimondi, Lavinia; De Luca, Angela; Amodio, Nicola; Manno, Mauro; Raccosta, Samuele; Taverna, Simona; Bellavia, Daniele; Naselli, Flores; Fontana, Simona; Schillaci, Odessa; Giardino, Roberto; Fini, Milena; Tassone, Pierfrancesco; Santoro, Alessandra; De Leo, Giacomo; Giavaresi, Gianluca; Alessandro, Riccardo

    2015-05-30

    Bone disease is the most frequent complication in multiple myeloma (MM) resulting in osteolytic lesions, bone pain, hypercalcemia and renal failure. In MM bone disease the perfect balance between bone-resorbing osteoclasts (OCs) and bone-forming osteoblasts (OBs) activity is lost in favour of OCs, thus resulting in skeletal disorders. Since exosomes have been described for their functional role in cancer progression, we here investigate whether MM cell-derived exosomes may be involved in OCs differentiation. We show that MM cells produce exosomes which are actively internalized by Raw264.7 cell line, a cellular model of osteoclast formation. MM cell-derived exosomes positively modulate pre-osteoclast migration, through the increasing of CXCR4 expression and trigger a survival pathway. MM cell-derived exosomes play a significant pro-differentiative role in murine Raw264.7 cells and human primary osteoclasts, inducing the expression of osteoclast markers such as Cathepsin K (CTSK), Matrix Metalloproteinases 9 (MMP9) and Tartrate-resistant Acid Phosphatase (TRAP). Pre-osteoclast treated with MM cell-derived exosomes differentiate in multinuclear OCs able to excavate authentic resorption lacunae. Similar results were obtained with exosomes derived from MM patient's sera. Our data indicate that MM-exosomes modulate OCs function and differentiation. Further studies are needed to identify the OCs activating factors transported by MM cell-derived exosomes.

  3. 黄芪有效部位对化疗性贫血小鼠细胞因子IL-2、 IL-4、 IL-6的影响%Effect of Effective Parts of Astragalus on Cytokines IL-2 IL-4and IL-6 of Chemotherapy-induced Anemic Mice

    Institute of Scientific and Technical Information of China (English)

    陈晶晶; 范颖; 张红梅; 林庶茹

    2011-01-01

    目的:探讨黄芪有效部位对化疗性贫血模型小鼠脾组织IL-2、IL-4、IL-6的影响.方法:采用ELISA 法测定脾组织IL-2、IL-4、IL-6含量.结果:模型组脾组织IL-2、IL-4含量下降,IL-6含量升高,均有统计学意义(P<0.05).各治疗组对化疗所致骨髓抑制性贫血的脾组织IL-2、IL-4、IL-6具有不同程度的调控作用.结论:IL-2、IL-4、IL-6与环磷酰胺所致骨髓抑制性贫血的免疫失调有关;黄芪及其有效部位对环磷酰胺引起的骨髓抑制性贫血引发的免疫损伤具有保护作用,并对IL-2、IL-4、IL-6的生成具有不同的调节机制.%Objective: To investigate effect of effective parts of astragalus on cytokines IL-2,IL-4 and IL-6 of chemotherapy-induced anemic mice. Methods : Enzyme-linked immunosorbent assay(ELISA) was applied to measure the content of IL-2, EL-4 and IL-6 in spleen. Results: In spleen of the model group, the content of IL-2 and IL-4 decreased, the content of IL-6 increased, both with significances in statistics(P<0.05). In each treated group, IL-2, IL-4 and IL-6 in spleen of chemotherapy-induced myelosuppression anemic mice are regulated in varying degree. Conclusion: There are relationship between immune disorders caused by cyclophosphamide-induced myelosuppression anemia and the content of IL-2, IL-4 and IL-6. Astragalus and its effective parts can protect immune injury caused by cyclophosphamide-induced myelosuppression anemia, also regulate the production of IL-2, IL-4 and IL-6 in different metabolism.

  4. Clinical significance of changes of serum IGF-II, IL-2 and SOD levels after treatment in pediatric patients with bronchial pneumonia

    International Nuclear Information System (INIS)

    Objective: To investigate the clinical significance of changes of serum IGF-II, IL-2 and SOD levels after treatment in pediatric patients with bronchial pneumonia. Methods: Serum IGF-II, IL-2 and SOD (with RIA) levels were measured in 33 pediatric patients with bronchial pneumonia both before and after treatment as well as in 35 controls. Results: Before treatment, serum IGF-II levels in the patients were significantly higher than those in controls (P0.05). Conclusion: Changes of serum IGF-II, IL-2 and SOD levels both before and after treatment could reflect the diseases status of the patients as well as the progress of diseases, and might be of prognostic importance in pediatric patients with bronchial pneumonia. (authors)

  5. Guanine nucleotide exchange factor αPIX leads to activation of the Rac 1 GTPase/glycogen phosphorylase pathway in interleukin (IL)-2-stimulated T cells

    DEFF Research Database (Denmark)

    Llavero, Francisco; Urzelai, Bakarne; Osinalde, Nerea;

    2015-01-01

    . More specifically, αPIX, a known guanine nucleotide exchange factor for the small GTPases of the Rho family, preferentially Rac 1, mediates PYGM activation in Kit 225 T cells stimulated with IL-2. Using directed mutagenesis, phosphorylation of αPIX Rho-GEF serines 225 and 488 is required for activation...... of the Rac 1/PYGM pathway. IL-2-stimulated serine phosphorylation was corroborated in Kit 225 T cells cultures. A parallel pharmacological and genetic approach identified PKCθ as the serine/threonine kinase responsible for αPIX serine phosphorylation. The phosphorylated state of αPIX was required to...... activate first Rac 1 and subsequently PYGM. These results demonstrate that the IL-2 receptor activation, among other early events, leads to activation of PKCθ. To activate Rac 1 and consequently PYGM, PKCθ phosphorylates αPIX in T cells. The biological significance of this PKCθ/αPIX/Rac 1 GTPase...

  6. Interleukin-2 (rIL-2)-induced lymphokine-activated killer (LAK) cells and their precursors express the VGO1 antigen

    International Nuclear Information System (INIS)

    Precursor and effector cells of recombinant interleukin-2 (r-IL-2)-induced lymphokine-activated killer (LAK) activity were investigated for their expression of VGO1. Peripheral blood lymphocytes (PBL) from normal donors were purified and separated in a FACS 420 into VGO1+- and VGO1- cell fractions before and after culture for 96 hr with 100 U/ml of r-IL-2. Their lytic activity against K 562 and Daudi cells was measured in a 51Cr release assay. The majority, if not all, of the LAK effector and precursor cells was VGO1+ lymphocytes. The expression of VGO1 by LAK precursor cells remained stable under the culture conditions used in our experiments. VGO1- lymphocytes cultured with r-IL-2 demonstrated neither LAK-induced activity nor expression of VGO1 antigen

  7. Clinical significance of determination of serum IL-2, IL-6 and GM-CSF levels after treatment in pediatric patients with bronchial asthma

    International Nuclear Information System (INIS)

    Objective: To explore the clinical significance of changes of serum IL-2, IL-6 and GM-CSF levels after treatment in pediatric patients with bronchial asthma. Methods: Serum levels of IL-2, IL-6 and GM-CSF were measured with RIA in 36 pediatric patients with bronchiol asthma and 30 controls. Results: Before treatment, the serum levels of IL-6, GM-CSF were significantly higher in the patients than those in controls (P0.05), Serum IL-2 levels were negatively correlated with the IL-6 and GM-CSF levels (r=-0.5846, -0.6018, P<0.01). Conclusion: These cytokines participated in the pathogenesis of bronchial asthma in pediatric patients. Mornitoring the changes of their serum levels was helpful for the management of the diseases. (authors)

  8. 松子壳多糖体外诱生小鼠IL-2和TNF-α的研究%Research on Polysaccharide from Pine Seed Shell Inducing Rat IL-2 andTNF-α in Vitro

    Institute of Scientific and Technical Information of China (English)

    刘中禄; 吕铁钢; 张永亮

    2010-01-01

    @@ 松子壳多糖(Pine nut shell polysaccharide,PSP)是从松子壳(松塔)中提取的酸性多糖成分,其在抗肿瘤、抗病毒和免疫促进剂作用方面研究较多(Sak-agami等,1999;Harada等,1991;吕永俊等,2008),我们在前期试验证实了松子壳多糖对小鼠脾T、B淋巴细胞转化、巨噬细胞的吞噬功能、NK细胞杀伤作用均有明显的促进作用,但PSP对细胞冈子的影响未见报道.本次试验进一步考察松子壳多糖对小鼠体外诱生IL-2及TNF-α分泌的影响,旨在为松子壳多糖的深入开发利用奠定一定的实验基础.

  9. Immunosuppressive effect of the anti-IL-2-receptor monoclonal antibody, AMT-13, on organ-cultured fetal pancreas allograft survival

    Energy Technology Data Exchange (ETDEWEB)

    Burkhardt, K.; Loughnan, M.S.; Diamantstein, T.; Mandel, T.E.

    1988-11-01

    Recently, prolongation of cardiac allograft survival in mice was reported using a rat anti-IL-2R mAb (AMT-13). However, its immunosuppressive action in vivo, alone and in combination with other immunosuppressants, and its effect on other organ transplants has not been extensively studied. We grafted cultured fetal pancreas from CBA (H-2k) donors to Balb/c (H-2d) mice. Recipients were treated with 10 consecutive daily injections each of 20 micrograms AMT-13 only, or with an additional mild immunosuppression of 350 rads irradiation. Control groups received rat immunoglobulin or 350 rads irradiation. Graft survival and the phenotype of infiltrating cells were assessed histologically and immunocytochemically on days 12, 17, and 21, and soluble IL-2R levels were measured in the serum with a quantitative ELISA in all recipients. Two of five grafts in the AMT-13-treated group had islets on day 12 posttransplantation despite lymphocytic infiltration in all grafts, while at this time all grafts of rat Ig treated control mice were completely rejected with only scar tissue and a few lymphocytes remaining. Additional immunosuppression with 350 rads irradiation had a marked additive effect with AMT-13. Soluble IL-2R levels in the serum of untreated recipients were not elevated compared with normal serum levels, but recipients injected with AMT-13 had multifold increased soluble IL-2R levels. The percentage of IL-2R+ cells in the grafts of AMT-13-treated animals was either normal (less than 5%) or increased (20%) in the additionally irradiated mice, providing strong evidence that the immunosuppressive effect of AMT-13 is not due to a depletion of activated IL-2R+ lymphocytes.

  10. Maturation of Stem Cell-Derived Beta-cells Guided by the Expression of Urocortin 3

    OpenAIRE

    van der Meulen, Talitha; Huising, Mark O.

    2014-01-01

    Type 1 diabetes (T1D) is a devastating disease precipitated by an autoimmune response directed at the insulin-producing beta-cells of the pancreas for which no cure exists. Stem cell-derived beta-cells show great promise for a cure as they have the potential to supply unlimited numbers of cells that could be derived from a patient's own cells, thus eliminating the need for immunosuppression. Current in vitro protocols for the differentiation of stem cell-derived beta-cells can successfully ge...

  11. Polymorphic variant at the IL2 region is associated with type 1 diabetes and may affect serum levels of interleukin-2

    OpenAIRE

    Fichna, Marta; Żurawek, Magdalena; Fichna, Piotr; Ziółkowska-Suchanek, Iwona; Januszkiewicz, Danuta; Nowak, Jerzy

    2013-01-01

    Polymorphic variants at the interleukin-2 (IL2) locus affect the risk of several autoimmune disorders. Our aim was to evaluate the association of the four IL2 polymorphisms (rs6822844, rs6534349, rs2069762 and rs3136534) with type 1 diabetes (T1D) in the Polish population, and to correlate them with the serum interleukin-2 levels. 543 unrelated T1D patients and 706 healthy control subjects were enrolled. The minor T allele at rs6822844 was significantly less frequent in T1D compared to contro...

  12. Lysis of pig endothelium by IL-2 activated human natural killer cells is inhibited by swine and human major histocompatibility complex (MHC) class I gene products.

    Science.gov (United States)

    Itescu, S; Artrip, J H; Kwiatkowski, P A; Wang, S F; Minanov, O P; Morgenthau, A S; Michler, R E

    1997-01-01

    We have previously described a form of xenograft rejection, mediated by natural killer (NK) cells, occurring in pig-to-primate organ transplants beyond the period of antibody-mediated hyperacute rejection. In this study, two distinct NK activation pathways were identified as mechanisms of pig aortic endotheliual cell (PAEC) lysis by human NK cells. Using an antibody-dependent cellular cytotoxicity (ADCC) assay, a progressive increase in human NK lysis of PAEC was observed following incubation with human IgG at increasing serum titer. In the absence of IgG, a second mechanism of PAEC lysis by human NK cells was observed following activation with IL-2. IL-2 activation of human NK cells increased lysis of PAEC by over 3-fold compared with ADCC. These results indicate that IL-2 activation of human NK cells induces significantly higher levels of lytic activity than does conventional ADCC involving IgG and FcRIII. We next investigated the role of MHC class I molecules in the regulation of NK lysis following IL-2 activation. PAEC expression of SLA class I molecules was increased by up to 75% by treatment with human TNFa. Following treatment with TNFa at 1 u/ml, IL-2 activated human NK lysis of PAEC was inhibited at every effector:target (E:T) ratio tested. Maximal effect occurred at an E:T ratio of 10:1, with TNFa inhibiting specific lysis by 59% (p < 0.01). Incubation with an anti-SLA class I Mab, but not IgG isotype control, abrogated the protective effects of TNFa on NK lysis of PAEC, suggesting direct inhibitory effects of SLA class I molecules on human NK function. To investigate whether human MHC class I molecules might have similar effects on human NK lysis of PAEC, further experiments were performed using a soluble peptide derived from the alpha-helical region of HLA-B7. Incubation with the HLA-B7 derived peptide significantly reduced the IL-2 activated NK lytic activity against PAEC in a dose-dependent fashion. Maximal effect occurred at a concentration of 10 mg

  13. Changes in peripheral blood level of regulatory T cells in patients with malignant melanoma during treatment with dendritic cell vaccination and low-dose IL-2

    DEFF Research Database (Denmark)

    Bjoern, J; Brimnes, M K; Andersen, M H;

    2011-01-01

    In this study, changes in peripheral blood regulatory T cell (Treg) levels were evaluated in 46 progressive patients with melanoma treated with a dendritic cell-based vaccine and concomitant low-dose IFN-a and IL-2. The regulatory subset of CD4 T cells, characterized by CD25(high) , was prospecti......In this study, changes in peripheral blood regulatory T cell (Treg) levels were evaluated in 46 progressive patients with melanoma treated with a dendritic cell-based vaccine and concomitant low-dose IFN-a and IL-2. The regulatory subset of CD4 T cells, characterized by CD25(high) , was...

  14. Changes in peripheral blood level of regulatory T cells in patients with malignant melanoma during treatment with dendritic cell vaccination and low-dose IL-2

    DEFF Research Database (Denmark)

    Bjoern, J; Brimnes, M K; Andersen, M H;

    2011-01-01

    In this study, changes in peripheral blood regulatory T cell (Treg) levels were evaluated in 46 progressive patients with melanoma treated with a dendritic cell-based vaccine and concomitant low-dose IFN-α and IL-2. The regulatory subset of CD4 T cells, characterized by CD25(high) , was prospecti......In this study, changes in peripheral blood regulatory T cell (Treg) levels were evaluated in 46 progressive patients with melanoma treated with a dendritic cell-based vaccine and concomitant low-dose IFN-α and IL-2. The regulatory subset of CD4 T cells, characterized by CD25(high) , was...

  15. Kinetics of human T-cell expression of LFA-1, IL-2 receptor, and ICAM-1 following antigenic stimulation in vitro

    DEFF Research Database (Denmark)

    Hviid, L; Felsing, A; Theander, T G

    1993-01-01

    in vitro is paralleled by differential kinetics in the expression of the T-cell adhesion and activation antigens leukocyte function-associated antigen 1 (LFA-1; CD11a/CD18), interleukin-2 receptor (IL-2R; CD25), and intercellular adhesion molecule 1 (ICAM-1; CD54). Furthermore, the changes in expression...... prestimulation levels, and CD25 expression was decreasing. This indicates that T-cell expression of all the 3 surface antigens examined is reversible. While this is in agreement with previous reports of the expression kinetics of IL-2R and ICAM-1, this is the first report indicating that the regulation of T...

  16. Observation and study OH IL-1β,IL-2 and IL-10 in serum of inbred HFJ rats%近交系HFJ大鼠血清中IL-1β、IL-2和IL-10观察研究

    Institute of Scientific and Technical Information of China (English)

    段肖翠; 吕占军; 蔡月花; 王秀芳; 侯洁; 刘军须; 刘福英; 谢英

    2011-01-01

    目的 比较观察本实验室培育的近交系HFJ大鼠和Wistar大鼠血清中IL-1β、IL-2和IL-10的含量.方法 HFJ大鼠和远交群Wistar大鼠各10只,应用ELISA法分别检测其血清IL-1β、IL-2和IL-10水平.结果 HFJ大鼠血清中IL-1β、IL-2的含量与Wistar大鼠相比差异无统计学意义(P>0.05),HFJ大鼠血清中IL-10的含量高于Wistar大鼠组,差异有统计学意义(P<0.05).且HFJ大鼠的IL-1β、IL-2和IL-10总体离散度明显小于Wistar大鼠组.结论 HFJ大鼠血清IL-10高于Wistar大鼠且近交系HFJ大鼠的IL-1β、IL-2和IL-10,测定值较为均一,离散度均低于Wistar大鼠.%Objective To comparative study the contents of IL - 1β, IL -2 and IL -10 in serum of Wistar rats and high fecundity inbred HFJ rats bred in our lab. Metbod Selecting 10 cases of HFJ rats and 10 cases of Wistar rats, and using ELISA to detected IL - 1β. IL -2 and IL - 10 in serum of them. Results There was no difference between Wistar group and HFJ group at the level of IL - 1β and IL - 2 (P > 0. 05) . The level of IL - 10 in HFJ group was higher than that in Wistar group (P < 0. 05) . The overall dispersion of IL - 1β. IL -2 and IL - 10 in HFJ rats group was significantly lower than Wistar rats group. Conlcusions IL - 10 in HFJ was higher than Wistar rats, and the measured values of IL -1β IL - 2 and IL - 10 were more relatively uniform in HFJ, and the dispersion of IL - 1β, IL - 2 and IL - 10 in HFJ rats group was lower than Wistar rats group.

  17. Fibrinogen-like protein 2/fibroleukin prothrombinase contributes to tumor hypercoagulability via IL-2 and IFN-γ

    Institute of Scientific and Technical Information of China (English)

    Kai Su; Fang Chen; Wei-Ming Yan; Qi-Li Zeng; Li Xu; Dong Xi; Bin Pi; Xiao-Ping Luo; Qin Ning

    2008-01-01

    AIM: To examine the role of Fibrinogen-like protein 2 (fgl2)/fibroleukin in tumor development. Fgl2 has been reported to play a vital role in the pathogenesis in MHV-3 (mouse hepatitis virus) induced fulminant and severe hepatitis, spontaneous abortion, allo- and xeno- graft rejection by mediating "immune coagulation".METHODS: Tumor tissues from 133 patients with six types of distinct cancers and the animal tumor tissues from human hepatocellular carcinoma (HCC) model on nude mice (established from high metastasis HCC cell line MHCC97LM6) were obtained.RESULTS: Hfgl2 was detected in tumor tissues from 127 out of 133 patients as well as tumor tissues collected from human HCC nude mice. Hfgl2 was highly expressed both in cancer cells and interstitial inflammatory cells including macrophages, NK cells, and CD8+ T lymphocytes and vascular endothelial cells. Hfgl2 mRNA was localized in cells that expressed hfgl2 protein. Fibrin (nogen) co-localization with hfgl2 expression was determined by dual immunohistochemical staining. In vitro, IL-2 and IFN-γ increased hfgl2 mRNA by 10-100 folds and protein expression in both THP-1 and HUVEC cell lines. One-stage clotting assays demonstrated that THP-1 and HUVEC cells expressing hfgl2 had increased procoagulant activity following cytokines stimulation.CONCLUSION: The hfgl2 contributes to the hypercoagulability in cancer and may induce tumor angiogenesis and metastasis via cytokine induction.

  18. Cancer Cell-derived Exosomes Induce Mitogen-activated Protein Kinase-dependent Monocyte Survival by Transport of Functional Receptor Tyrosine Kinases.

    Science.gov (United States)

    Song, Xiao; Ding, Yanping; Liu, Gang; Yang, Xiao; Zhao, Ruifang; Zhang, Yinlong; Zhao, Xiao; Anderson, Gregory J; Nie, Guangjun

    2016-04-15

    Tumor-associated macrophages (TAM) play pivotal roles in cancer initiation and progression. Monocytes, the precursors of TAMs, normally undergo spontaneous apoptosis within 2 days, but can subsist in the inflammatory tumor microenvironment for continuous survival and generation of sufficient TAMs. The mechanisms underlying tumor-driving monocyte survival remain obscure. Here we report that cancer cell-derived exosomes were crucial mediators for monocyte survival in the inflammatory niche. Analysis of the survival-promoting molecules in monocytes revealed that cancer cell-derived exosomes activated Ras and extracellular signal-regulated kinases in the mitogen-activated protein kinase (MAPK) pathway, resulting in the prevention of caspase cleavage. Phosphorylated receptor tyrosine kinases (RTKs), such as phosphorylated epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 (HER-2), were abundantly expressed in cancer cell-derived exosomes. Knock-out of EGFR or/and HER-2, or alternatively, inhibitors against their phosphorylation significantly disturbed the exosome-mediated activation of the MAPK pathway, inhibition of caspase cleavage, and increase in survival rate in monocytes. Moreover, the deprived survival-stimulating activity of exosomes due to null expression of EGFR and HER-2 could be restored by activation of another RTK, insulin receptor. Overall, our study uncovered a mechanism of tumor-associated monocyte survival and demonstrated that cancer cell-derived exosomes can stimulate the MAPK pathway in monocytes through transport of functional RTKs, leading to inactivation of apoptosis-related caspases. This work provides insights into the long sought question on monocyte survival prior to formation of plentiful TAMs in the tumor microenvironment.

  19. Polyphosphate induces matrix metalloproteinase-3-mediated proliferation of odontoblast-like cells derived from induced pluripotent stem cells

    International Nuclear Information System (INIS)

    Inorganic polyphosphate [Poly(P)] may represent a physiological source of phosphate and has the ability to induce bone differentiation in osteoblasts. We previously reported that cytokine-induced matrix metalloproteinase (MMP)-3 accelerates the proliferation of purified odontoblast-like cells. In this study, MMP-3 small interfering RNA (siRNA) was transfected into odontoblast-like cells derived from induced pluripotent stem cells to investigate whether MMP-3 activity is induced by Poly(P) and/or is associated with cell proliferation and differentiation into odontoblast-like cells. Treatment with Poly(P) led to an increase in both cell proliferation and additional odontoblastic differentiation. Poly(P)-treated cells showed a small but significant increase in dentin sialophosphoprotein (DSPP) and dentin matrix protein-1 (DMP-1) mRNA expression, which are markers of mature odontoblasts. The cells also acquired additional odontoblast-specific properties including adoption of an odontoblastic phenotype typified by high alkaline phosphatase (ALP) activity and a calcification capacity. In addition, Poly(P) induced expression of MMP-3 mRNA and protein, and increased MMP-3 activity. MMP-3 siRNA-mediated disruption of the expression of these effectors potently suppressed the expression of odontoblastic biomarkers ALP, DSPP, and DMP-1, and blocked calcification. Interestingly, upon siRNA-mediated silencing of MMP-3, we noted a potent and significant decrease in cell proliferation. Using specific siRNAs, we revealed that a unique signaling cascade, Poly(P)→MMP-3→DSPP and/or DMP-1, was intimately involved in the proliferation of odontoblast-like cells. - Highlights: • Polyphosphate increases proliferation of iPS cell-derived odontoblast-like cells. • Polyphosphate-induced MMP-3 results in an increase of cell proliferation. • Induced cell proliferation involves MMP-3, DSPP, and/or DMP-1 sequentially. • Induced MMP-3 also results in an increase of odontoblastic

  20. Polyphosphate induces matrix metalloproteinase-3-mediated proliferation of odontoblast-like cells derived from induced pluripotent stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Ozeki, Nobuaki; Hase, Naoko; Yamaguchi, Hideyuki; Hiyama, Taiki; Kawai, Rie [Department of Endodontics, School of Dentistry, Aichi Gakuin University, 2-11 Suemori-dori, Chikusa-ku, Nagoya, Aichi 464-8651 (Japan); Kondo, Ayami [Department of Medicinal Biochemistry, School of Pharmacy, Aichi Gakuin University, 1-100 Kusumoto, Chikusa-ku, Nagoya 464-8650 (Japan); Nakata, Kazuhiko [Department of Endodontics, School of Dentistry, Aichi Gakuin University, 2-11 Suemori-dori, Chikusa-ku, Nagoya, Aichi 464-8651 (Japan); Mogi, Makio, E-mail: makio@dpc.agu.ac.jp [Department of Medicinal Biochemistry, School of Pharmacy, Aichi Gakuin University, 1-100 Kusumoto, Chikusa-ku, Nagoya 464-8650 (Japan)

    2015-05-01

    Inorganic polyphosphate [Poly(P)] may represent a physiological source of phosphate and has the ability to induce bone differentiation in osteoblasts. We previously reported that cytokine-induced matrix metalloproteinase (MMP)-3 accelerates the proliferation of purified odontoblast-like cells. In this study, MMP-3 small interfering RNA (siRNA) was transfected into odontoblast-like cells derived from induced pluripotent stem cells to investigate whether MMP-3 activity is induced by Poly(P) and/or is associated with cell proliferation and differentiation into odontoblast-like cells. Treatment with Poly(P) led to an increase in both cell proliferation and additional odontoblastic differentiation. Poly(P)-treated cells showed a small but significant increase in dentin sialophosphoprotein (DSPP) and dentin matrix protein-1 (DMP-1) mRNA expression, which are markers of mature odontoblasts. The cells also acquired additional odontoblast-specific properties including adoption of an odontoblastic phenotype typified by high alkaline phosphatase (ALP) activity and a calcification capacity. In addition, Poly(P) induced expression of MMP-3 mRNA and protein, and increased MMP-3 activity. MMP-3 siRNA-mediated disruption of the expression of these effectors potently suppressed the expression of odontoblastic biomarkers ALP, DSPP, and DMP-1, and blocked calcification. Interestingly, upon siRNA-mediated silencing of MMP-3, we noted a potent and significant decrease in cell proliferation. Using specific siRNAs, we revealed that a unique signaling cascade, Poly(P)→MMP-3→DSPP and/or DMP-1, was intimately involved in the proliferation of odontoblast-like cells. - Highlights: • Polyphosphate increases proliferation of iPS cell-derived odontoblast-like cells. • Polyphosphate-induced MMP-3 results in an increase of cell proliferation. • Induced cell proliferation involves MMP-3, DSPP, and/or DMP-1 sequentially. • Induced MMP-3 also results in an increase of odontoblastic

  1. Analysis of Cell-Derived Microparticles with Highly Precise Nanotechnological Methods

    DEFF Research Database (Denmark)

    Cherré, Solène; Østergaard, Ole; Heegaard, Niels H.H.;

    2014-01-01

    Cell-derived microparticles have gained a broad interest in the past years. Being released by blood cells upon activation or induction of apoptosis, they have a great potential as novel diagnostic markers and their investigation can bring new knowledge into the pathogenesis of various diseases. H...

  2. A novel mechanism of bacterial toxin transfer within host blood cell-derived microvesicles.

    Directory of Open Access Journals (Sweden)

    Anne-lie Ståhl

    2015-02-01

    Full Text Available Shiga toxin (Stx is the main virulence factor of enterohemorrhagic Escherichia coli, which are non-invasive strains that can lead to hemolytic uremic syndrome (HUS, associated with renal failure and death. Although bacteremia does not occur, bacterial virulence factors gain access to the circulation and are thereafter presumed to cause target organ damage. Stx was previously shown to circulate bound to blood cells but the mechanism by which it would potentially transfer to target organ cells has not been elucidated. Here we show that blood cell-derived microvesicles, shed during HUS, contain Stx and are found within patient renal cortical cells. The finding was reproduced in mice infected with Stx-producing Escherichia coli exhibiting Stx-containing blood cell-derived microvesicles in the circulation that reached the kidney where they were transferred into glomerular and peritubular capillary endothelial cells and further through their basement membranes followed by podocytes and tubular epithelial cells, respectively. In vitro studies demonstrated that blood cell-derived microvesicles containing Stx undergo endocytosis in glomerular endothelial cells leading to cell death secondary to inhibited protein synthesis. This study demonstrates a novel virulence mechanism whereby bacterial toxin is transferred within host blood cell-derived microvesicles in which it may evade the host immune system.

  3. Chemokine stromal cell-derived factor 1alpha activates basophils by means of CXCR4

    DEFF Research Database (Denmark)

    Jinquan, T; Jacobi, H H; Jing, C;

    2000-01-01

    The CXC chemokine receptor 4 (CXCR4) is predominantly expressed on inactivated naive T lymphocytes, B lymphocytes, dendritic cells, and endothelial cells. CXC chemokine stromal cell-derived factor 1alpha (SDF-1alpha) is the only known ligand for CXCR4. To date, the CXCR4 expression and function o...

  4. Construction of recombinant pPIC9K-IL-2 and expression in Pichia pastoris%pPIC9 K-IL-2重组质粒的构建及其在毕赤酵母中的表达

    Institute of Scientific and Technical Information of China (English)

    刘斌; 祁光宇; 陈晓宇; 王宇; 牟克斌; 黄银君; 刘学荣

    2014-01-01

    A pair of primers was designed to clone IL-2 gene according to the Ovis aries interleukin-2 ( IL-2)sequence and the recombinant expressed vector pPIC9K-IL-2 was constructed.The recombinane plas-mid was transformed into Pichia pastoris GS1 1 5 by electroporation.The transformants were selected with MD culture plate and identified by PCR.The multicopy recombinant P.pastoris strain was selected by G418 (4 g/L)resistance.The result showed that IL-2 gene was integrated with chromosome of P.pastoris by PCR identification.The expressed product had a molecular weight of 1 8 ku band by SDS-PAGE analy-sis.The recombinant P.pastoris GS1 1 5 was successfully constructed,which provided the basis for research and development of a novel cytokine adj uvant.%根据 Ovis aries interleukin (IL-2)序列设计1对引物,用 PCR法扩增的目的基因进行克隆,将测序正确的IL-2基因片段和表达载体 pPIC9K 同时双酶切后相连,构建 pPIC9K-IL-2重组表达载体。将线性化的载体pPIC9K-IL-2电转化毕赤酵母 GS115,对重组酵母转化子经 MD平板筛选和 PCR 分析鉴定后,用 G418(4 g/L)筛选到多拷贝菌株,进行不同条件下甲醇诱导表达。结果表明:经 PCR鉴定 IL-2基因已整合到酵母基因组中,表达产物经 SDS-PAGE电泳分析发现相对分子质量约为18 ku目的蛋白表达条带,本研究成功构建了高效表达重组羊IL-2毕赤酵母菌株,为新型细胞因子佐剂的研发奠定了基础。

  5. Expression patterns of IL-2 and IFN-γ of activated γδT cells from human peripheral blood%人外周血γδT细胞激活后IL-2和IFN-γ表达的特点

    Institute of Scientific and Technical Information of China (English)

    陈礼文; 朱安友; 李柏青

    2005-01-01

    目的:探讨人外周血γδT细胞在不同刺激剂激活后产生Th1型细胞因子IL-2和IFN-γ的不同特点.方法:健康成人外周血单个核细胞(PBMC)用佛波醇酯(PMA)、离子霉素(ionomycin)和抗CD28单抗(mAb)以不同组合刺激6~8 h后;用流式细胞术胞内细胞因子检测法测定γδT细胞和αβT细胞IL-2和IFN-γ的表达;或用结核杆菌抗原(Mtb-Ag)刺激γδT细胞加IL-2扩增3~8天后检测γδT细胞IL-2的表达.结果:PBMC经PMA+ionomycin刺激后,与αβT细胞中产生IL-2细胞(14.0%)相比,γδT细胞中极少数细胞(1.1%)表达IL-2;但分泌IFN-γ细胞在γδT细胞中(56.1%)明显多于αβT细胞(8.5%).如加抗CD28 mAb联合刺激后,IL-2产生细胞在αβT细胞中明显增加,但在γδT细胞中未见增加;而IFN-γ表达细胞在两类细胞中均有明显增加;PBMC用Mtb-Ag激活后扩增3天后在γδT细胞中开始检测到IL-2表达,第5~6天时达高峰,第8天时逐渐下降.结论:多克隆短时刺激对PBMC中γδT细胞表达IFN-γ能力强于αβT细胞,但产生IL-2很少;Mtb-Ag能诱导γδT细胞表达IL-2,但过程较慢.

  6. Effects of vitamin E on the proliferation and collagen synthesis of rat hepatic stellate cells treated with IL-2 or TNF-α

    Institute of Scientific and Technical Information of China (English)

    展玉涛; 王宇; 魏来; 陈红松

    2003-01-01

    Objective To study the effects of vitamin E on the proliferation and collagen synthesis of rat hepatic stellate cells treated with interleukin-2 (IL-2 ) or tumor necrosis factor-α (TNF-α).Methods Hepatic stellate cells were isolated from male Sprague-Dawley rats by using modified Friedman's method. Using the isolated cells cultured and treated with IL-2 or TNF-α, we studied the effects of vitamin E on their proliferation and collagen synthesis through an 3 H-thymidine and 3 H-proline incorporation assay, as well as through observation of these cells under a contrary phase microscope. Results Adding IL-2 increased the both proliferation and collagen synthesis of hepatic stellate cells. Their proliferation was also increased by the addition of TNF-α, although it decreased collagen synthesis. Vitamin E had marked inhibitory effects on the ability of cells treated with IL-2 or TNF-α to reproduce or synthesize collagen.Conclusion Vitamin E can inhibit the proliferation and collagen synthesis of hepatic stellate cells. It is possible that vitamin E affects liver fibrosis through these activities.

  7. Microbial infection-induced expansion of effector T cells overcomes the suppressive effects of regulatory T cells via an IL-2 deprivation mechanism.

    Science.gov (United States)

    Benson, Alicia; Murray, Sean; Divakar, Prashanthi; Burnaevskiy, Nikolay; Pifer, Reed; Forman, James; Yarovinsky, Felix

    2012-01-15

    Foxp3(+) regulatory T (Treg) cells are a critical cell population that suppresses T cell activation in response to microbial and viral pathogens. We identify a cell-intrinsic mechanism by which effector CD4(+) T cells overcome the suppressive effects of Treg cells in the context of three distinct infections: Toxoplasma gondii, Listeria monocytogenes, and vaccinia virus. The acute responses to the parasitic, bacterial, and viral pathogens resulted in a transient reduction in frequency and absolute number of Treg cells. The infection-induced partial loss of Treg cells was essential for the initiation of potent Th1 responses and host protection against the pathogens. The observed disappearance of Treg cells was a result of insufficiency in IL-2 caused by the expansion of pathogen-specific CD4(+) T cells with a limited capacity of IL-2 production. Exogenous IL-2 treatment during the parasitic, bacterial, and viral infections completely prevented the loss of Treg cells, but restoration of Treg cells resulted in a greatly enhanced susceptibility to the pathogens. These results demonstrate that the transient reduction in Treg cells induced by pathogens via IL-2 deprivation is essential for optimal T cell responses and host resistance to microbial and viral pathogens. PMID:22147768

  8. Steroid-withdrawal at 3 days after renal transplantation with anti-IL-2 receptor alpha therapy: a prospective, randomized, multicenter study.

    NARCIS (Netherlands)

    Meulen, C.G. ter; Riemsdijk, I.C. van; Hene, R.J.; Christiaans, M.H.; Borm, G.F.; Gelder, T. van; Hilbrands, L.B.; Weimar, W.; Hoitsma, A.J.

    2004-01-01

    Steroids have been included in most immunosuppressive regimens after renal transplantation, but are feared for their side-effects. We conducted a prospective multicenter study to investigate whether it is feasible to withdraw steroids early after transplantation with the use of anti-IL-2Ralpha induc

  9. Local IL2 and IL12 treatment of Bovine Ocular Squamous Cell Carcinoma (BOSCC) and Bovine Vulval Papilloma and Carcinoma Complex (BVPCC) in Cattle in Zimbabwe

    NARCIS (Netherlands)

    Stewart, R.J.E.

    2007-01-01

    Effect of local IL-2 application on bovine cancer In tropical countries there is an increased prevalence of two important cancers in cattle: Bovine Ocular Squamous Cell Carcinoma (BOSCC) and Bovine Vulval Papilloma Carcinoma Complex (BVPCC). Both cancers are associated with increased annual hours of

  10. Acute exhaustive exercise regulates IL-2, IL-4 and MyoD in skeletal muscle but not adipose tissue in rats

    Directory of Open Access Journals (Sweden)

    Seelaender Marília

    2011-06-01

    Full Text Available Abstract Background The purpose of this study was to evaluate the effect of exhaustive exercise on proteins associated with muscle damage and regeneration, including IL-2, IL-4 and MyoD, in extensor digitorum longus (EDL and soleus muscles and mesenteric (MEAT and retroperitoneal adipose tissues (RPAT. Methods Rats were killed by decapitation immediately (E0 group, n = 6, 2 (E2 group, n = 6 or 6 (E6 group, n = 6 hours after the exhaustion protocol, which consisted of running on a treadmill at approximately 70% of VO2max for fifty minutes and then at an elevated rate that increased at one m/min every minute, until exhaustion. Results The control group (C group, n = 6 was not subjected to exercise. IL-2 protein expression increased at E0 in the soleus and EDL; at E2, this cytokine returned to control levels in both tissues. In the soleus, IL-2 protein expression was lower than that in the control at E6. IL-4 protein levels increased in EDL at E6, but the opposite result was observed in the soleus. MyoD expression increased at E6 in EDL. Conclusion Exhaustive exercise was unable to modify IL-2 and IL-4 levels in MEAT and RPAT. The results show that exhaustive exercise has different effects depending on which muscle is analysed.

  11. Constitutive release of IFNγ and IL2 from peripheral blood mononuclear cells of rhesus macaques (Macaca mulatta) infected with simian T-lymphotropic virus type 1.

    Science.gov (United States)

    Yee, JoAnn L; Montiel, Nestor A; Ardeshir, Amir; Ardeshr, Amir; Lerche, Nicholas W

    2013-01-01

    Simian T-cell lymphotropic viruses (STLV), the nonhuman primate counterparts of human T-cell lymphotropic viruses (HTLV), are endemic in many populations of African and Asian monkeys and apes. Although an etiologic link between STLV1 infection and lymphoproliferative disorders such as malignant lymphomas has been suggested in some nonhuman primate species, most STLV infections are inapparent, and infected animals remain clinically healthy. The retroviral transactivator, tax, is well known to increase transcription of viral and cellular genes, resulting in altered cytokine profiles. This study compared the cytokine profiles of peripheral blood mononuclear cell (PBMC) cultures from 25 STLV1-seropositive rhesus macaques (Macaca mulatta) with those of age- and sex-matched seronegative controls. IFNγ, TNFα, IL10, and IL2 levels in unstimulated PBMC culture supernatants were measured at 24, 48, and 72 h by using enzyme immunoassays. IFNγ concentrations were found significantly higher in the supernatants of PBMC cultures of seropositive monkeys as compared with seronegative controls. In addition, although IL2 concentrations were not significantly elevated in the supernatants of PBMC cultures of all seropositive monkeys as compared with all seronegative controls, IL2 levels were increased in a subset of 5 pairs. Increased constitutive cytokine release occurred in the absence of spontaneous proliferation. The increased constitutive release of IFNγ and IL2 suggests that STLV1 alters immune functions in infected but clinically healthy rhesus macaques and further characterizes STLV1 infection of rhesus macaques as a potential model for human HTLV1 infection. PMID:24326227

  12. Constitutive Release of IFNγ and IL2 from Peripheral Blood Mononuclear Cells of Rhesus Macaques (Macaca mulatta) Infected with Simian T-Lymphotropic Virus Type 1

    Science.gov (United States)

    Yee, JoAnn L; Montiel, Nestor A; Ardeshr, Amir; Lerche, Nicholas W

    2013-01-01

    Simian T-cell lymphotropic viruses (STLV), the nonhuman primate counterparts of human T-cell lymphotropic viruses (HTLV), are endemic in many populations of African and Asian monkeys and apes. Although an etiologic link between STLV1 infection and lymphoproliferative disorders such as malignant lymphomas has been suggested in some nonhuman primate species, most STLV infections are inapparent, and infected animals remain clinically healthy. The retroviral transactivator, tax, is well known to increase transcription of viral and cellular genes, resulting in altered cytokine profiles. This study compared the cytokine profiles of peripheral blood mononuclear cell (PBMC) cultures from 25 STLV1-seropositive rhesus macaques (Macaca mulatta) with those of age- and sex-matched seronegative controls. IFNγ, TNFα, IL10, and IL2 levels in unstimulated PBMC culture supernatants were measured at 24, 48, and 72 h by using enzyme immunoassays. IFNγ concentrations were found significantly higher in the supernatants of PBMC cultures of seropositive monkeys as compared with seronegative controls. In addition, although IL2 concentrations were not significantly elevated in the supernatants of PBMC cultures of all seropositive monkeys as compared with all seronegative controls, IL2 levels were increased in a subset of 5 pairs. Increased constitutive cytokine release occurred in the absence of spontaneous proliferation. The increased constitutive release of IFNγ and IL2 suggests that STLV1 alters immune functions in infected but clinically healthy rhesus macaques and further characterizes STLV1 infection of rhesus macaques as a potential model for human HTLV1 infection. PMID:24326227

  13. 重组腺病毒人白细胞介素2的质量分析%QUALITY CONTROL FOR RECOMBINANT ADENOVIRUS-IL2

    Institute of Scientific and Technical Information of China (English)

    林建伟; 王军志; 饶春明

    2002-01-01

    目的通过对重组腺病毒人白细胞介素2(Adv-IL2)的质量研究和实验方法参数的选择,建立Adv-IL2的质量控制标准.方法选用CPE法测定病毒滴度;将病毒转染Hela细胞,表达出IL-2,分别用MTT法和ELISA法测定IL-2的活性和表达量.建立了UV和IE-HPLC系统分析纯度;琼脂糖凝胶电泳检查基因组分子量和基因组酶切图谱;PCR法鉴定病毒特征基因E2B区、IL-2表达基因盒和外源因子(复制型病毒RCV,HIV,HBV和HCV);按照的有关规定进行了其他项目检查.结果测定的重组腺病毒人白细胞介素2原液滴度达3×1010 pfu*mL-1.IL-2表达活性达700 U*mL-1,表达量约25 ng*mL-1.制品的A260 nm/A280 nm为1.23;IE-HPLC纯度检查大于98%.基因组分子量和基因组酶切图谱以及病毒特征基因E2B区和IL-2表达基因检查结果均在理论值范围;复制型病毒RCV,HIV,HBV和HCV检测均呈阴性;其他项目均符合规定.结论建立了重组腺病毒人白细胞介素2的质量标准,并可有效地控制制品的质量.

  14. 首发精神分裂症患者使用阿立哌唑后血清IL-2、IL-4水平变化的探讨%Explore the level changes of IL-2,IL-4 in the first-episode schizophrenia after the treatment of arip-iprazole

    Institute of Scientific and Technical Information of China (English)

    刘倩倩; 李亚飞; 朱祥路; 蒋天玉

    2014-01-01

    Objective To explore the differences of serum IL-2 ,IL-4 in the first-episode schizo-phrenia and healthy controls were explored ,and to compare the changes of symptoms before and after aripiprazole treatment and the changes of serum IL-2 ,IL-4 .Methods Serum of IL-2 ,IL-4 was exam-ined with Flow Cytometry in 35 healthy volunteers and 35 first episode patients .The symptoms of pa-tients were evaluated with Positive and Negative Syndrome Scale .Results There were no statistical sig-nificantly differents in the serum of IL-2 ,IL-4 in the first-episode schizophrenia than normal .controls (P>0 .05) .The serum levels of IL-4 was lower in patients with first-episode schizophrenia after aripi-prazole treatment (P<0 .01) .IL-2 and IL-4 levels were increased in positive symptoms of schizophre-nia patients before aripiprazole treatment (positive symptoms) than normal controls (P<0 .05) .IL-2 and IL-4 levels were different in positive symptoms of schizophrenia patients before and after aripi-prazole treatment (P<0 .05) .Conclusion The patients with schizophrenia have immune dysfunction ;Aripiprazole of antipsychotics have lowered the level of IL-2 ,IL-4 and positive symptoms also im-proved .Conclusion.%目的:探讨首发精神分裂症患者血清细胞因子IL-2、IL-4与正常人的差异,比较分析首发精神分裂症患者经过阿立哌唑治疗前后症状改变及细胞因子IL-2、IL-4的变化。方法选择35例首发精神分裂症患者作为研究组,35例健康志愿者作为对照组,通过流式细胞学技术测定血清标本中IL-2、IL-4的水平,用PANSS量表评定精神症状。结果(1)首发精神分裂症患者IL-2、IL-4水平与正常对照组相比,差异无统计学意义(P>0.05)。(2)首发精神分裂症患者阿立哌唑治疗后较治疗前IL-4水平降低,差异有统计学意义(P<0.01)。(3)首发精神分裂症阳性症状患者血清IL-2、IL-4水平在治疗前均高于对照组(P<0.05

  15. Transgene Reactivation in Induced Pluripotent Stem Cell Derivatives and Reversion to Pluripotency of Induced Pluripotent Stem Cell-Derived Mesenchymal Stem Cells.

    Science.gov (United States)

    Galat, Vasiliy; Galat, Yekaterina; Perepitchka, Mariana; Jennings, Lawrence J; Iannaccone, Philip M; Hendrix, Mary J C

    2016-07-15

    Induced pluripotent stem cells (iPSCs) have enormous potential in regenerative medicine and disease modeling. It is now felt that clinical trials should be performed with iPSCs derived with nonintegrative constructs. Numerous studies, however, including those describing disease models, are still being published using cells derived from iPSCs generated with integrative constructs. Our experimental work presents the first evidence of spontaneous transgene reactivation in vitro in several cellular types. Our results show that the transgenes were predominantly silent in parent iPSCs, but in mesenchymal and endothelial iPSC derivatives, the transgenes experienced random upregulation of Nanog and c-Myc. Additionally, we provide evidence of spontaneous secondary reprogramming and reversion to pluripotency in mesenchymal stem cells derived from iPSCs. These findings strongly suggest that the studies, which use cellular products derived from iPSCs generated with retro- or lentiviruses, should be evaluated with consideration of the possibility of transgene reactivation. The in vitro model described here provides insight into the earliest events of culture transformation and suggests the hypothesis that reversion to pluripotency may be responsible for the development of tumors in cell replacement experiments. The main goal of this work, however, is to communicate the possibility of transgene reactivation in retro- or lenti-iPSC derivatives and the associated loss of cellular fidelity in vitro, which may impact the outcomes of disease modeling and related experimentation. PMID:27193052

  16. 重组小鼠白介素-2基因在真核细胞中的转染表达%THE EXPRESSION OF RECOMBINANT mIL-2 GENE IN TRANSFECTED EUKARYOTIC CELLS

    Institute of Scientific and Technical Information of China (English)

    路晓辉; 杜军; 曾军; 李晓玲; 徐珍霞; 蔡绍晖

    2006-01-01

    目的:在构建重组真核表达载体pIRESneo 2/mIL-2的基础上,建立能够持续稳定表达mIL-2的哺乳类工程细胞.方法:运用分子克隆技术,将由RT-PCR获得的mIL-2 cDNA片断插入真核表达质粒pIRESneo2构建成mIL-2重组表达载体pIRESneo2/mIL-2.通过脂质体转染法将pIRESneo2/mIL-2导入C2C12细胞.转染后第30天,用Western blots检测mIL-2表达情况.结果:经DNA测序证明mIL-2 cDNA片断插入方向和碱基组成顺序均准确无误,Western blots检测转染真核重组表达载体pIRESneo 2/mIL-2的C2C12细胞系表达mIL-2.结论:利用pIRESneo2/mIL-2构建的真核表达载体在C2C12细胞系中能够持续稳定表达mIL-2.

  17. 尖锐湿疣组织IL-2、IL-4、IL-10、IL-18表达的意义

    Institute of Scientific and Technical Information of China (English)

    胡凯; 程方雄; 陈蓓; 韩林; 马鸣

    2012-01-01

    目的:探讨尖锐湿疣(CA)患者治疗前、后血清中白细胞介素(IL)-2、IL-4、IL-10和IL-18的水平变化在CA免疫发病机制中的作用.方法:用酶联免疫吸附法(ELISA)分别检测30例CA患者在治疗前和治疗后3个月未复发时血清IL-2、IL-4 、ID-10和IL-18的水平,并与20例正常人作比较.结果:结果CA患者治疗前血清中IL-2水平明显低于正常对照组(P<0.01),IL-4、IL-10和IL-18水平明显高于正常对照组(P<0.05);治疗后3个月末血清IL-2、IL-4、IL-10、IL-18的水平同正常对照组相比,差异无统计学意义(均P>0.05).结论:人乳头瘤病毒(HPV)感染可造成患者细胞免疫功能异常,IL-2、IL-4、IL-10和IL 18在CA免疫发病机制中可能起重要作用.

  18. 尖锐湿疣组织IL-2、IL-4、IL-1O、IL-18的表达

    Institute of Scientific and Technical Information of China (English)

    胡凯; 程方雄; 陈蓓; 韩林; 马鸣

    2012-01-01

    目的:探讨尖锐湿疣(CA)患者治疗前、后血清中白介素2(IL-2)、白介素4(IL-4)、白介素10(IL-10)和白介素18(IL-18)的水平变化在CA免疫发病机制中的作用.方法:用酶联免疫吸附法(ELISA)分别检测30例CA患者在治疗前和治疗后3个月未复发时血清IL-2、IL-4、IL-10和IL-18的水平,并与20例正常人作比较.结果:CA患者治疗前血清中IL2水平明显低于正常对照组(P>0.01),IL-4、IL-10和IL-18水平明显高于正常对照组(P>0.05);治疗后3个月末血清IL-2、IL-4、IL-10、IL-18的水平同正常对照组相比,差异无统计学意义(P均>0.05).结论:人乳头瘤病毒(HPV)感染可造成患者细胞免疫功能异常,IL2、IL-4、IL-10和IL-18在CA免疫发病机制中可能起重要作用.

  19. Development of an antibody to bovine IL-2 reveals multifunctional CD4 T(EM cells in cattle naturally infected with bovine tuberculosis.

    Directory of Open Access Journals (Sweden)

    Adam O Whelan

    Full Text Available Gaining a better understanding of the T cell mechanisms underlying natural immunity to bovine tuberculosis would help to identify immune correlates of disease progression and facilitate the rational design of improved vaccine and diagnostic strategies. CD4 T cells play an established central role in immunity to TB, and recent interest has focussed on the potential role of multifunctional CD4 T cells expressing IFN-γ, IL-2 and TNF-α. Until now, it has not been possible to assess the contribution of these multifunctional CD4 T cells in cattle due to the lack of reagents to detect bovine IL-2 (bIL-2. Using recombinant phage display technology, we have identified an antibody that recognises biologically active bIL-2. Using this antibody, we have developed a polychromatic flow cytometric staining panel that has allowed the investigation of multifunctional CD4 T-cells responses in cattle naturally infected with M. bovis. Assessment of the frequency of antigen specific CD4 T cell subsets reveals a dominant IFN-γ(+IL-2(+TNF-α(+ and IFN-γ(+ TNF-α(+ response in naturally infected cattle. These multifunctional CD4 T cells express a CD44(hiCD45RO(+CD62L(lo T-effector memory (T(EM phenotype and display higher cytokine median fluorescence intensities than single cytokine producers, consistent with an enhanced 'quality of response' as reported for multifunctional cells in human and murine systems. Through our development of these novel immunological bovine tools, we provide the first description of multifunctional T(EM cells in cattle. Application of these tools will improve our understanding of protective immunity in bovine TB and allow more direct comparisons of the complex T cell mediated immune responses between murine models, human clinical studies and bovine TB models in the future.

  20. Recombinant human B7-H4 expressed in Escherichia coli inhibits T lymphocyte proliferation and IL-2 secretion in vitro

    Institute of Scientific and Technical Information of China (English)

    Yi-xiang MAO; Xue-guang ZHANG; Yong-Jing CHEN; Van GE; Hong-bing MA; Jian-feng YU; Hong-ya WU; Yu-min HU; Qin WANG; Qin SHI

    2006-01-01

    Aim: To explore the biofunctions of human B7-H4 generated from prokaryotic system. Methods: The gene of human B7-H4 extracellular region (IgⅤ-like and IgC-like domains) was obtained by PCR from human cDNA FLJ22418 and then inserted into the prokaryotic expression vector pGEX-5X-3 expressing glutathione. r-transferase (GST) fusion protein. After being identified by restriction enzyme digestion and sequencing, the recombinant vector was transferred into host strain E coli BL21-RIL(DE3). A 47 kDa fusion protein (GST/hB7-H4) was induced by isopropyl-beta-D-thiogalactopyranoside (IPTG) and purified by standard methods reported in the prokaryotic system. The inhibitory effect of GST/hB7-H4 on proliferation of T cells was observed in vitro by CD3mAb activated T-cell cultur-ing system and [3H]-thymidine incorporation assay. The concentrations of interleukin-2 and iterferon-g in the supernatants of T cells were determined by ELISA. Results: We successfully constructed the method for high-level expression and purification of the hB7-H4 extracellular domain as GST fusion protein from E coli. The GST/hB7-H4 fusion protein produced in bacteria had obvious biological activity to inhibit T-lymphocyte proliferation and IL-2 secretion. Conclusion: The prokaryote expression system could be used to generate hB7-H4 protein with natural spatial conformations and biological functions, which provided an efficient and economical way for the preparation of this target protein.

  1. IL-2R common gamma-chain is epigenetically silenced by nucleophosphin-anaplastic lymphoma kinase (NPM-ALK) and acts as a tumor suppressor by targeting NPM-ALK.

    Science.gov (United States)

    Zhang, Qian; Wang, Hong Yi; Liu, Xiaobin; Bhutani, Gauri; Kantekure, Kanchan; Wasik, Mariusz

    2011-07-19

    Anaplastic lymphoma kinase (ALK), physiologically expressed only by certain neural cells, becomes highly oncogenic, when aberrantly expressed in nonneural tissues as a fusion protein with nucleophosphin (NPM) and other partners. The reason why NPM-ALK succeeds in transforming specifically CD4(+) T lymphocytes remains unknown. The IL-2R common γ-chain (IL-2Rγ) is shared by receptors for several cytokines that play key roles in the maturation and growth of normal CD4(+) T lymphocytes and other immune cells. We show that IL-2Rγ expression is inhibited in T-cell lymphoma cells expressing NPM-ALK kinase as a result of DNA methylation of the IL-2Rγ gene promoter. IL-2Rγ promoter methylation is induced in malignant T cells by NPM-ALK. NPM-ALK acts through STAT3, a transcription factor that binds to the IL-2Rγ gene promoter and enhances binding of DNA methyltransferases (DNMTs) to the promoter. In addition, STAT3 suppresses expression of miR-21, which selectively inhibits DNMT1 mRNA expression. Reconstitution of IL-2Rγ expression leads to loss of the NPM-ALK protein and, consequently, apoptotic cell death of the lymphoma cells. These results demonstrate that the oncogenic tyrosine kinase NPM-ALK induces epigenetic silencing of the IL-2Rγ gene and that IL-2Rγ acts as a tumor suppressor by reciprocally inhibiting expression of NPM-ALK.

  2. Vaccination with a plasmid DNA encoding HER-2/neu together with low doses of GM-CSF and IL-2 in patients with metastatic breast carcinoma: a pilot clinical trial

    Directory of Open Access Journals (Sweden)

    Knutson Keith L

    2010-06-01

    Full Text Available Abstract Background Adjuvant trastuzumab (Herceptin treatment of breast cancer patients significantly improves their clinical outcome. Vaccination is an attractive alternative approach to provide HER-2/neu (Her2-specific antibodies and may in addition concomitantly stimulate Her2-reactive T-cells. Here we report the first administration of a Her2-plasmid DNA (pDNA vaccine in humans. Patients and Methods The vaccine, encoding a full-length signaling-deficient version of the oncogene Her2, was administered together with low doses of GM-CSF and IL-2 to patients with metastatic Her2-expressing breast carcinoma who were also treated with trastuzumab. Six of eight enrolled patients completed all three vaccine cycles. In the remaining two patients treatment was discontinued after one vaccine cycle due to rapid tumor progression or disease-related complications. The primary objective was the evaluation of safety and tolerability of the vaccine regimen. As a secondary objective, treatment-induced Her2-specific immunity was monitored by measuring antibody production as well as T-cell proliferation and cytokine production in response to Her2-derived antigens. Results No clinical manifestations of acute toxicity, autoimmunity or cardiotoxicity were observed after administration of Her2-pDNA in combination with GM-CSF, IL-2 and trastuzumab. No specific T-cell proliferation following in vitro stimulation of freshly isolated PBMC with recombinant human Her2 protein was induced by the vaccination. Immediately after all three cycles of vaccination no or even decreased CD4+ T-cell responses towards Her2-derived peptide epitopes were observed, but a significant increase of MHC class II restricted T-cell responses to Her2 was detected at long term follow-up. Since concurrent trastuzumab therapy was permitted, λ-subclass specific ELISAs were performed to specifically measure endogenous antibody production without interference by trastuzumab. Her2-pDNA vaccination

  3. NOD/scid IL-2Rgnull mice: a preclinical model system to evaluate human dendritic cell-based vaccine strategies in vivo

    Directory of Open Access Journals (Sweden)

    Spranger Stefani

    2012-02-01

    Full Text Available Abstract Background To date very few systems have been described for preclinical investigations of human cellular therapeutics in vivo. However, the ability to carry out comparisons of new cellular vaccines in vivo would be of substantial interest for design of clinical studies. Here we describe a humanized mouse model to assess the efficacy of various human dendritic cell (DC preparations. Two reconstitution regimes of NOD/scid IL2Rgnull (NSG mice with adult human peripheral blood mononuclear cells (PBMC were evaluated for engraftment using 4-week and 9-week schedules. This led to selection of a simple and rapid protocol for engraftment and vaccine evaluation that encompassed 4 weeks. Methods NSG recipients of human PBMC were engrafted over 14 days and then vaccinated twice with autologous DC via intravenous injection. Three DC vaccine formulations were compared that varied generation time in vitro (3 days versus 7 days and signals for maturation (with or without Toll-like receptor (TLR3 and TLR7/8 agonists using MART-1 as a surrogate antigen, by electroporating mature DC with in vitro transcribed RNA encoding full length protein. After two weekly vaccinations, the splenocyte populations containing human lymphocytes were recovered 7 days later and assessed for MART-1-specific immune responses using MHC-multimer-binding assays and functional assessment of specific killing of melanoma tumor cell lines. Results Human monocyte-derived DC generated in vitro in 3 days induced better MART-1-specific immune responses in the autologous donor T cells present in the humanized NSG mice. Moreover, consistent with our in vitro observations, vaccination using mature DC activated with TLR3 and TLR7/8 agonists resulted in enhanced immune responses in vivo. These findings led to a ranking of the DC vaccine effects in vivo that reflected the hierarchy previously found for these mature DC variations in vitro. Conclusions This humanized mouse model system enables

  4. 热化疗治疗非小细胞肺癌对IL-2、IL-6、IL-8、IL-10及TNF的影响%Effect of thermochemotherapy on levels of IL-2 ,IL-6 ,IL-8 ,IL-10 and TNF in non-small cell lung cancer

    Institute of Scientific and Technical Information of China (English)

    李新民; 尉继伟; 刘治邦; 刘建国

    2014-01-01

    目的 探讨热化疗治疗非小细胞肺癌(NSCLC)对白细胞介素-2(IL-2)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、白细胞介素-10(IL-10)及肿瘤坏死因子(TNF)的影响.方法 回顾性分析大同大学附属医院2004年10月至2013年1月收治的134例NSCLC患者,进行热疗联合NP方案化疗(NVB+ DDP),并分别在治疗前、治疗3个周期后、治疗6个周期后、治疗结束后3个月对患者血清中IL-2、IL-6、IL-8、IL-10及TNF的变化进行监测.结果 热化疗3个周期后,IL-2、TNF水平逐渐增高,明显高于治疗前;IL-6、IL-8和IL-10水平明显低于化疗前.热化疗6个周期后,血IL-6、IL-8、IL-10、IL-2和TNF都有所下降,其中IL-2、TNF浓度显著低于化疗3个周期后的水平.热化疗结束后3个月,血IL-6、IL-8、IL-10水平继续下降;而IL-2、TNF浓度逐渐增高,低于化疗3个周期后的水平,但高于治疗前水平.结论 热化疗治疗NSCLC对IL-2、TNF水平有明显的增高作用,而对IL-6、IL-8、IL-10有降低作用.%Objective To investigate the effect of hot therapy on interleukin-2 (IL-2),interleukin-6 (IL-6),interlerukin-8 (IL-8),interleukin-10 (IL-10) and tumor necrosis factor (TNF) in nonsmall cell lung cancer(NSCLC).Methods One hundred and thirty-four patients with NSCLC,admitted to our hospital from October 2004 to January 2013,received hot therapy and vinorelbine plus cisplatin (NP) chemotherapy.The IL-6,IL-8,IL-10 and TNF level before therapy,after 3 cycles,after 6 cycles and 3 months after chemotherapy were observed.Results IL-2 and TNF levels increased gradually after 3 cycles of hot therapy,and were significant higher than those before therapy.Compared to before therapy,IL-6,IL-8 and IL-10 levels significantly decreased.IL-6,IL-8,IL-10,IL-2 and TNF levels all decreased at 6 months after hot therapy.IL-2 and TNF levels were significant lowered than those of 3 cycles after chemotherapy.IL-6,IL-8 and IL-10 continued to decrease 3 months after the end of

  5. Detection and its clinical significance of IL-2, IL-10 and TNF-α in the sera of patients with early symptomatic syphilis%早期显性梅毒患者血清IL-2、IL-10和TNF-α检测及其临床意义

    Institute of Scientific and Technical Information of China (English)

    胡晓燕; 聂芳

    2012-01-01

    目的 探讨白细胞介素-2(IL-2)、白细胞介素-10(IL-10)和肿瘤坏死因子-α(TNF-α)在早期显性梅毒发病机制中的作用.方法 采用双抗体夹心酶联免疫吸附试验(DAbS-ELISA)检测53例早期显性梅毒患者(其中一期梅毒25例、二期梅毒28例)和65名正常人血清IL-2、IL-10和TNF-α水平.结果 早期显性梅毒患者血清IL-2、IL-10和TNF-α水平明显高于正常对照组(P<0.01);一期梅毒患者血清IL-2和TNF-α水平明显高于二期梅毒患者(P<0.01),血清IL-10水平则低于二期梅毒患者(P<0.01);早期显性梅毒患者中IL-2与IL-10呈负相关(r=-0.760,P =0.000),与TNF-α呈正相关(r=0.633,P=0.000),而IL-10与INF-α无明显相关性(r=-0.063,P =0.575).结论 IL-2、IL-10和TNF-α均参与了梅毒的发病,IL-10表达上调可能是二期梅毒发生辅助性T(Th)1/Th2细胞免疫应答失衡的主要原因之一.%Objective To investigate the role of interleukin-2 (IL-2), interleukin-10( IL-10) and tumor necrosis factor-alpha (TNF-α) in the pathogenesis of early symptomatic syphilis. Methods Double-antibody sandwich enzyme-linked immunosorbent assay(DAbS-ELISA) was used to detect the serum levels of IL-2,IL-10 and TNF-α in 53 patients with early symptomatic syphilis (25 cases with primary syphilis and 28 cases with secondary syphilis) and 65 healthy subjects. Results Serum levels of IL-2, IL-10 and TNF-α in early symptomatic syphilis group were significantly higher than those in control group (P <0. 01). Serum levels of IL-2 and TNF-α in primary syphilis group were significantly higher than those in secondary syphilis group (P < 0. 01). Serum level of IL-10 in secondary syphilis group was significantly higher than that in primary syphilis group (P<0.01). IL-2 and IL-10 were negatively correlated in early symptomatic syphilis group (r = - 0. 760, P = 0. 000). IL-2 and TNF-α were positively correlated ( r = 0. 633, P = 0.000), and IL-10 and TNF-α were not correlated(r = -0

  6. Imaging of inflamed carotid artery atherosclerotic plaques with the use of {sup 99m}Tc-HYNIC-IL-2 scintigraphy in end-stage renal disease patients

    Energy Technology Data Exchange (ETDEWEB)

    Opalinska, Marta; Pach, Dorota; Sowa-Staszczak, Anna; Glowa, Boguslaw; Hubalewska-Dydejczyk, Alicja [Jagiellonian University Medical School, Nuclear Medicine Unit, Department of Endocrinology, Cracow (Poland); Stompor, Tomasz [University of Warmia and Mazury in Olsztyn, Department of Nephrology, Hypertensiology and Internal Medicine, Faculty of Medicine, Olsztyn (Poland); Mikolajczak, Renata; Garnuszek, Piotr; Maurin, Michal; Karczmarczyk, Urszula [National Centre for Nuclear Research Radioisotope Centre POLATOM, Otwock (Poland); Fedak, Danuta [Jagiellonian University Medical School, Clinical Biochemistry, Cracow (Poland); Krzanowski, Marcin; Sulowicz, Wladyslaw [Jagiellonian University Medical School, Department of Nephrology, Cracow (Poland); Rakowski, Tomasz [Jagiellonian University Medical School, 2nd Department of Cardiology, Institute of Cardiology, Cracow (Poland)

    2012-04-15

    Identification of vulnerable plaques remains crucial for better cardiovascular risk assessment. At least 20% of inflammatory cells within unstable (vulnerable) plaques comprise T lymphocytes, which contain receptors for interleukin-2 (IL-2); those receptors can be identified by scintigraphy with radiolabelled IL-2.The aim of this study was to identify the ''inflamed'' (vulnerable) plaques by scintigraphy using IL-2 labelled with {sup 99m}Tc in the selected, high cardiovascular risk group of end-stage renal disease (ESRD) patients. A total of 28 patients (18 men, 10 women, aged 55.2 {+-} 9.6 years, 17 on peritoneal dialysis, 11 on haemodialysis) underwent common carotid artery (CCA) scintigraphy with the use of {sup 99m}Tc-hydrazinonicotinamide (HYNIC)-IL-2. In all cases, ultrasound examination of the CCA was performed and levels of selected proinflammatory factors, atherogenic markers and calcium-phosphate balance parameters were measured. Finally, the target to non-target (T/nT) ratio of IL-2 uptake in atherosclerotic plaques with intima-media thickness (IMT), classic cardiovascular risk factors and concentrations of the measured factors were compared. Increased {sup 99m}Tc-HYNIC-IL-2 uptake in atherosclerotic plaques in 38/41 (91%) cases was detected. The median T/nT ratio of focal {sup 99m}Tc-HYNIC-IL-2 uptake in atherosclerotic plaques was 2.35 (range 1.23-3.63). The mean IMT value on the side of plaques assessed by scintigraphy was 0.79 {+-} 0.18 mm (median 0.8, range 0.5-1.275). Correlations between T/nT ratio and homocysteine (R = 0.22, p = 0.037), apolipoprotein B (apoB) (R = 0.31, p = 0.008), apoB to apoA-I ratio (R = 0.29, p = 0.012) and triglyceride concentration (R = 0.26, p = 0.021) were detected. A lower T/nT ratio in patients with better parameters of nutritional status (haemoglobin, albumin, adiponectin) in comparison with patients with worse nutritional parameters (3.20 {+-} 0.5 vs 2.16 {+-} 0.68, p = 0.025) was revealed as well

  7. Clinical Significance of Changes of Serum IL-2,IL-6 and Gastrin Levels Before and After Treatment in Patients with Chronic Eczema%慢性湿疹患者治疗前后血清IL-2、IL-6和Gas检测的临床意义

    Institute of Scientific and Technical Information of China (English)

    牟勖东; 任虹

    2009-01-01

    目的:探讨慢性湿疹患者治疗前后血清IL-2、IL-6和胃泌素(Gas)水平的变化及意义.方法:应用放射免疫分析对38例慢性湿疹患者进行治疗前后血清IL-2、IL-6和Gas检测,并与35名正常健康人作比较.结果:在治疗前血清IL-2水平非常显著地低于正常人组(P<0.01),而IL-6和Gas则显著地高于正常人组(P<0.01),经治疗1个月后,除IL-6、Gas水平与正常人组比较无显著性差异外(P>0.05),IL-2与正常人组比较仍有显著性差异(P<0.05).结论:检测慢性湿疹患者血清IL-2、IL-6和Gas水平的变化对了解病情、指导临床实践有重要的临床价值.

  8. Clinical Significance of Measurement the Changes on Serum IL-2, IL-8, IL-18 and VEGF Levels After Treatment in Patients with Acute Cerebral Infarction%ACI患者治疗前后血清IL-2、IL-8、IL-18和VEGF检测的临床意义

    Institute of Scientific and Technical Information of China (English)

    张春雷; 章红梅

    2013-01-01

    目的:探讨了急性脑梗死(ACI)患者治疗前后血清IL-2、IL-8、IL-18和VEGF水平的变化及临床意义.方法:应用放射免疫分析、酶联法对33例ACI患者进行了治疗前后血清IL-2、IL-8、IL-18和VEGF检测,并与35名正常健康人作比较.结果:ACI患者在治疗前血清IL-8、IL-18和VEGF水平非常显著地高于正常人组(P<0.01),而血清IL-2水平又非常显著地低于正常人组(P<0.01),经治疗3个月后与正常人组比较仍有显著性差异(P<0.05).且血清IL-2水平与IL-8、IL-18和VEGF水平呈显著负相关(r=-0.4218、-0.4726、-0.5014,P<0.01).结论:ACI的发生、发展与血清IL-2、IL-8、IL-18和VEGF水平密切相关.

  9. 特发性血小板减少性紫癜患儿血清IL-6,sIL-2R的变化及意义%Changes of serum IL-6 and sIL-2R levels in patients with idiopathic thrombocytopenic purpura and their clinical significance

    Institute of Scientific and Technical Information of China (English)

    张国利; 张剑白; 蒋丽鑫

    2009-01-01

    Objective To study the changes and the clinical significance of serum IL-6 and sIL-2R levels in children with idiopathic thrombocytopenic purpura( ITP). Methods The levels of serum IL-6 and sIL-2R were measured in 37 ITP children with sandwich ELSIA as well as in 20 healthy children. At the same time, PAIb' s qualitative determination was checked in 25 ITP children. And the relationships of PLT counts,IL-6 and sIL-2R's level in ITP children' s peripheral blood were analyzed. Results The levels of ITP children' s IL-6 and sIL-2R were significantly higher than those of control group(P <0. 01). The levels of IL-6 and sIL-2R were dramatically higher in the group of ITP children before treatment responsive to after treatment(P <0.01 ). The levels of severe hemorrhage group' s IL-6, sIL-2R were significantly higher than those of hyporrhea group' s( P < 0.05 ). Nine of 25 ITP children' s PAIb was completely positive(9/25) ,the levels of ITP children's IL-6, sIL-2R in completely positive group were significantly higher than non-positive group' s( P < 0. 05 ). The levels of ITP children' s IL-6, sIL-2R ( before treating) were negative correlation with PLT counts. The level between ITP children' s IL-6 and sIL- 2R was positive correlation. Conclusion PIL-6 and sIL-2R participate in ITP' s procedure. The levels change with the pathogenetic condition and the course of disease. To sum up, cell mediated immunity plays a role in ITP' s procedure as well as humoral-mediated immunity.%目的 探讨特发性血小板减少性紫癜(ITP)患儿血清中细胞因子白介素6(IL-6)、可溶性白介素2受体(sIL-2R)的变化及临床意义.方法 采用双抗体夹心酶联免疫(ELISA)定量测定方法检测37例ITP患儿和20例正常健康儿童血清中IL-6、sIL-2R水平,同时对其中25例ITP患儿进行血小板相关抗体PAIb(IgG、IgA、IgM)的定性测定,并对ITP患儿外周血中血小板计数、血清IL-6、sIL-2R水平进行相关分析.结论 ITP患儿血清IL-6、sIL

  10. Microfluidic isolation of cancer-cell-derived microvesicles from hetergeneous extracellular shed vesicle populations.

    Science.gov (United States)

    Santana, Steven M; Antonyak, Marc A; Cerione, Richard A; Kirby, Brian J

    2014-12-01

    Extracellular shed vesicles, including exosomes and microvesicles, are disseminated throughout the body and represent an important conduit of cell communication. Cancer-cell-derived microvesicles have potential as a cancer biomarker as they help shape the tumor microenvironment to promote the growth of the primary tumor and prime the metastatic niche. It is likely that, in cancer cell cultures, the two constituent extracellular shed vesicle subpopulations, observed in dynamic light scattering, represent an exosome population and a cancer-cell-specific microvesicle population and that extracellular shed vesicle size provides information about provenance and cargo. We have designed and implemented a novel microfluidic technology that separates microvesicles, as a function of diameter, from heterogeneous populations of cancer-cell-derived extracellular shed vesicles. We measured cargo carried by the microvesicle subpopulation processed through this microfluidic platform. Such analyses could enable future investigations to more accurately and reliably determine provenance, functional activity, and mechanisms of transformation in cancer. PMID:25342569

  11. Glucose responsive insulin production from human embryonic germ (EG) cell derivatives

    International Nuclear Information System (INIS)

    Type 1 diabetes mellitus subjects millions to a daily burden of disease management, life threatening hypoglycemia and long-term complications such as retinopathy, nephropathy, heart disease, and stroke. Cell transplantation therapies providing a glucose-regulated supply of insulin have been implemented clinically, but are limited by safety, efficacy and supply considerations. Stem cells promise a plentiful and flexible source of cells for transplantation therapies. Here, we show that cells derived from human embryonic germ (EG) cells express markers of definitive endoderm, pancreatic and β-cell development, glucose sensing, and production of mature insulin. These cells integrate functions necessary for glucose responsive regulation of preproinsulin mRNA and expression of insulin C-peptide in vitro. Following transplantation into mice, cells become insulin and C-peptide immunoreactive and produce plasma C-peptide in response to glucose. These findings suggest that EG cell derivatives may eventually serve as a source of insulin producing cells for the treatment of diabetes

  12. Microencapsulation technology by nature: Cell derived extracellular vesicles with therapeutic potential.

    Science.gov (United States)

    Kittel, A; Falus, A; Buzás, E

    2013-06-01

    Cell derived extracellular vesicles are submicron structures surrounded by phospholipid bilayer and released by both prokaryotic and eukaryotic cells. The sizes of these vesicles roughly fall into the size ranges of microbes, and they represent efficient delivery platforms targeting complex molecular information to professional antigen presenting cells. Critical roles of these naturally formulated units of information have been described in many physiological and pathological processes. Extracellular vesicles are not only potential biomarkers and possible pathogenic factors in numerous diseases, but they are also considered as emerging therapeutic targets and therapeutic vehicles. Strikingly, current drug delivery systems, designed to convey therapeutic proteins and peptides (such as liposomes), show many similarities to extracellular vesicles. Here we review some aspects of therapeutic implementation of natural, cell-derived extracellular vesicles in human diseases. Exploration of molecular and functional details of extracellular vesicle release and action may provide important lessons for the design of future drug delivery systems.

  13. Education, tobacco smoking, alcohol consumption, and IL-2 and IL-6 gene polymorphisms in the survival of head and neck cancer

    Directory of Open Access Journals (Sweden)

    R.V.M. López

    2011-10-01

    Full Text Available The association of education, tobacco smoking, alcohol consumption, and interleukin-2 (IL-2 +114 and -384 and -6 (IL-6 -174 DNA polymorphisms with head and neck squamous cell carcinoma (HNSCC was investigated in a cohort study of 445 subjects. IL-2 and IL-6 genotypes were determined by real-time PCR. Cox regression was used to estimate hazard ratios (HR and 95% confidence intervals (95%CI of disease-specific survival according to anatomical sites of the head and neck. Mean age was 56 years and most patients were males (87.6%. Subjects with 5 or more years of schooling had better survival in larynx cancer. Smoking had no effect on HNSCC survival, but alcohol consumption had a statistically significant effect on larynx cancer. IL-2 gene +114 G/T (HR = 0.52; 95%CI = 0.15-1.81 and T/T (HR = 0.22; 95%CI = 0.02-3.19 genotypes were associated with better survival in hypopharynx cancer. IL-2 +114 G/T was a predictor of poor survival in oral cavity/oropharynx cancer and larynx cancer (HR = 1.32; 95%CI = 0.61-2.85. IL-2 -384 G/T was associated with better survival in oral cavity/oropharynx cancer (HR = 0.80; 95%CI = 0.45-1.42 and hypopharynx cancer (HR = 0.68; 95%CI = 0.21-2.20, but an inverse relationship was observed for larynx cancer. IL-6 -174 G/C was associated with better survival in hypopharynx cancer (HR = 0.68; 95%CI = 0.26-1.78 and larynx cancer (HR = 0.93; 95%CI = 0.42-2.07, and C/C reduced mortality in larynx cancer. In general, our results are similar to previous reports on the value of education, smoking, alcohol consumption, and IL-2 and IL-6 genetic polymorphisms for the prognosis of HNSCC, but the risks due to these variables are small and estimates imprecise.

  14. Serum concentrations of sIL-2R, IL-6, TGF-beta1, neopterin, and zinc in chronic hepatitis C patients treated with interferon-alpha.

    Science.gov (United States)

    Grüngreiff, K; Reinhold, D; Ansorge, S

    1999-12-01

    T lymphocytes and immunoregulatory cytokines play an important role in the host response to hepatitis C virus (HCV) infection. Zinc is required for a wide spectrum of immune functions, including T-cell activity. To determine the clinical significance of the cytokines sIL-2R, IL-6, TGF-beta1, neopterin, and of zinc in chronic heptatitis C virus (HCV) infection, we investigated their concentrations in the serum of 16 patients with chronic HCV infection before, during and at the end of therapy with interferon (IFN) alpha (Roferon A), and after 6 months follow-up. Elevated concentrations of sIL-2R, IL-6, TGF-beta1, and neopterin were found in the serum of all patients prior to therapy, as compared to healthy controls. sIL-2R patterns differed in responders and non-responders. While the mean concentration of sIL-2R (335.75 pg/ml) before therapy was about 40% higher in complete responders (n=4) than in controls (272.20 pg/ml), the mean concentration in non-responders (n=6) was 4-fold higher than in controls (1153.33 pg/ml). During therapy, sIL-2R levels in responders decreased by about 40%. Mean IL-6 concentrations in both complete and partial responders (n=6) decreased continuously during treatment, while mean concentrations in non-responders decreased for only a short time, and increased again after cessation of therapy. Mean levels of TGF-beta1 behaved similarly to those of IL-6. Only negligible differences in mean neopterin levels were found between responders and non-responders over the entire observation time. The mean serum zinc concentrations slightly decreased in all 3 patient groups, the greatest reduction occurring in 3 of the 4 responders. The present findings underscore the importance of the immune system in the pathogenesis of chronic HCV infection. Serum sIL-2R levels may be used as a serological marker of outcome following IFN-alpha treatment. PMID:10623433

  15. Efficacy of chimeric DNA vaccines encoding Eimeria tenella 5401 and chicken IFN-γ or IL-2 against coccidiosis in chickens.

    Science.gov (United States)

    Song, Xiaokai; Huang, Xinmei; Yan, Ruofeng; Xu, Lixin; Li, Xiangrui

    2015-09-01

    Chimeric DNA vaccines encoding Eimeria tenella (E. tenella) surface antigen 5401 were constructed and their efficacies against E. tenella challenge were studied. The open reading frame (ORF) of 5401 was cloned into the prokaryotic expression vector pGEX-4T2 to express the recombinant protein and the expressed recombinant protein was identified by Western blot. The ORF of 5401 and chicken cytokine gene IFN-γ or IL-2 were cloned into the eukaryotic expression vector pVAX1 consecutively to construct DNA vaccines pVAX-5401-IFN-γ, pVAX-5401-IL-2 and pVAX-5401. The expression of aim genes in vivo was detected by reverse transcription-polymerase chain reaction and Western blot. Fourteen-day-old chickens were inoculated twice at an interval of 7 days with 100 µg of plasmids pVAX-5401, pVAX-5401-IFN-γ and pVAX-5401-IL-2 or 200 µg of recombinant 5401 protein by leg intramuscular injection, respectively. Seven days after the second inoculation, all chickens except the unchallenged control group were challenged orally with 5 × 10(4) sporulated oocysts of E. tenella. Seven days after challenge, all chickens were weighted and slaughtered to determine the effects of immunization. The results showed the recombinant protein was about 90 kDa and reacted with antiserum against soluble sporozoites. The animal experiment showed that all the DNA vaccines pVAX-5401, pVAX-5401-IFN-γ or pVAX-5401-IL-2 and the recombinant 5401 protein could obviously alleviate body weight loss and cecal lesions as compared with non-vaccinated challenged control and empty vector pVAX1control. Furthermore, pVAX-5401-IFN-γ or pVAX-5401-IL-2 induced anti-coccidial index (ACI) of 180.01 or 177.24 which were significantly higher than that of pVAX-5401. The results suggested that 5401 was an effective candidate antigen for vaccine. This finding also suggested that chicken IFN-γ or IL-2 could effectively improve the efficacies of DNA vaccines against avian coccidiosis.

  16. Increase in IFNγ(-IL-2(+ cells in recent human CD4 T cell responses to 2009 pandemic H1N1 influenza.

    Directory of Open Access Journals (Sweden)

    Jason M Weaver

    Full Text Available Human CD4 T cell recall responses to influenza virus are strongly biased towards Type 1 cytokines, producing IFNγ, IL-2 and TNFα. We have now examined the effector phenotypes of CD4 T cells in more detail, particularly focusing on differences between recent versus long-term, multiply-boosted responses. Peptides spanning the proteome of temporally distinct influenza viruses were distributed into pools enriched for cross-reactivity to different influenza strains, and used to stimulate antigen-specific CD4 T cells representing recent or long-term memory. In the general population, peptides unique to the long-circulating influenza A/New Caledonia/20/99 (H1N1 induced Th1-like responses biased toward the expression of IFNγ(+TNFα(+ CD4 T cells. In contrast, peptide pools enriched for non-cross-reactive peptides of the pandemic influenza A/California/04/09 (H1N1 induced more IFNγ(-IL-2(+TNFα(+ T cells, similar to the IFNγ(-IL-2(+ non-polarized, primed precursor T cells (Thpp that are a predominant response to protein vaccination. These results were confirmed in a second study that compared samples taken before the 2009 pandemic to samples taken one month after PCR-confirmed A/California/04/09 infection. There were striking increases in influenza-specific TNFα(+, IFNγ(+, and IL-2(+ cells in the post-infection samples. Importantly, peptides enriched for non-cross-reactive A/California/04/09 specificities induced a higher proportion of Thpp-like IFNγ(-IL-2(+TNFα(+ CD4 T cells than peptide pools cross-reactive with previous influenza strains, which induced more Th1 (IFNγ(+TNFα(+ responses. These IFNγ(-IL-2(+TNFα(+ CD4 T cells may be an important target population for vaccination regimens, as these cells are induced upon infection, may have high proliferative potential, and may play a role in providing future effector cells during subsequent infections.

  17. 二十碳五烯酸影响T细胞膜脂肪微区域中IL-2受体的分布%EICOSAPENTAENOIC ACID ALTERS INTERLEUKIN-2 RECEPTOR DISTRIBUTION IN DETERGENT-INSOLUBLE MEMBRANE DOMAIN

    Institute of Scientific and Technical Information of China (English)

    李秋荣; 马健; 汪灏; 黎介寿

    2005-01-01

    目的:研究二十碳五烯酸(EPA)对T细胞膜脂肪微区域(lipid rafts)中IL-2受体(IL-2R)分布的影响.方法:EPA(20:5)处理Jurkat E6-1 T细胞为实验组,硬脂酸(18:0)处理为阴性对照.用流式细胞仪检测对T细胞表面分子CD25(IL-2α受体)表达的抑制作用.应用蛋白免疫印迹分析,化学发光法检测IL-2α受体所在的T细胞膜脂肪微区域分离组分.结果:用硬脂酸处理T细胞,细胞表面CD25阳性表达细胞为39.53%,不同浓度的EPA(5、12.5、25,50、75μmol/L)处理,CD25阳性表达细胞分别为36.12%、31.30%、23.59%、16.67%和11.65%,EPA可抑制T细胞表面分子CD25的表达.蛋白印迹分析确定IL-2α、IL-2Rβ和IL-2Rγc都存在于微区域组分中,EPA处理使部分IL-2Rα、IL-2Rβ和IL-2Rγc从膜脂肪微区域移位到可溶膜区域.结论:膜脂肪微区域为IL-2受体信号传导的功能性亚区域,EPA通过调节IL-2Rot、IL-2Rβ和IL-2Rγc在膜脂肪微区域的分布,使部分IL-2R从功能性脂肪微区域移位到非功能性可溶膜区域,而产生免疫抑制作用.

  18. Cytokine diversity in the Th1-dominated human anti-influenza response caused by variable cytokine expression by Th1 cells, and a minor population of uncommitted IL-2+IFNγ- Thpp cells.

    Directory of Open Access Journals (Sweden)

    Nan Deng

    Full Text Available Within overall Th1-like human memory T cell responses, individual T cells may express only some of the characteristic Th1 cytokines when reactivated. In the Th1-oriented memory response to influenza, we have tested the contributions of two potential mechanisms for this diversity: variable expression of cytokines by a uniform population during activation, or different stable subsets that consistently expressed subsets of the Th1 cytokine pattern. To test for short-term variability, in vitro-stimulated influenza-specific human memory CD4+ T cells were sorted according to IL-2 and IFNγ expression, cultured briefly in vitro, and cytokine patterns measured after restimulation. Cells that were initially IFNγ+ and either IL-2+ or IL-2- converged rapidly, containing similar proportions of IL-2-IFNγ+ and IL-2+IFNγ+ cells after culture and restimulation. Both phenotypes expressed Tbet, and similar patterns of mRNA. Thus variability of IL-2 expression in IFNγ+ cells appeared to be regulated more by short-term variability than by stable differentiated subsets. In contrast, heterogeneous expression of IFNγ in IL-2+ influenza-specific T cells appeared to be due partly to stable T cell subsets. After sorting, culture and restimulation, influenza-specific IL-2+IFNγ- and IL-2+IFNγ+ cells maintained significantly biased ratios of IFNγ+ and IFNγ- cells. IL-2+IFNγ- cells included both Tbetlo and Tbethi cells, and showed more mRNA expression differences with either of the IFNγ+ populations. To test whether IL-2+IFNγ-Tbetlo cells were Thpp cells (primed but uncommitted memory cells, predominant in responses to protein vaccines, influenza-specific IL-2+IFNγ- and IL-2+IFNγ+ T cells were sorted and cultured in Th1- or Th2-generating conditions. Both cell types yielded IFNγ-secreting cells in Th1 conditions, but only IL-2+IFNγ- cells were able to differentiate into IL-4-producing cells. Thus expression of IL-2 in the anti-influenza response may be

  19. Tumor-targeted delivery of IL-2 by NKG2D leads to accumulation of antigen-specific CD8+ T cells in the tumor loci and enhanced anti-tumor effects.

    Directory of Open Access Journals (Sweden)

    Tae Heung Kang

    Full Text Available Interleukin-2 (IL-2 has been shown to promote tumor-specific T-cell proliferation and differentiation but systemic administration of IL-2 results in significant toxicity. Therefore, a strategy that can specifically deliver IL-2 to the tumor location may alleviate concerns of toxicity. Because NKG2D ligands have been shown to be highly expressed in many cancer cells but not in healthy cells, we reason that a chimeric protein consisting of NKG2D linked to IL-2 will lead to the specific targeting of IL-2 to the tumor location. Therefore, we created chimeric proteins consisting of NKG2D linked to Gaussia luciferase (GLuc; a marker protein or IL-2 to form NKG2D-Fc-GLuc and NKG2D-Fc-IL2, respectively. We demonstrated that NKG2D linked to GLuc was able to deliver GLuc to the tumor location in vivo. Furthermore, we showed that TC-1 tumor-bearing mice intramuscularly injected with DNA encoding NKG2D-Fc-IL2, followed by electroporation, exhibited an increased number of luciferase-expressing E7-specific CD8+ T cells at the tumor location. More importantly, treatment with the DNA construct encoding NKG2D-Fc-IL2 significantly enhanced the therapeutic anti-tumor effects generated by intradermal vaccination with therapeutic HPV DNA in tumor-bearing mice. Therefore, by linking NKG2D to IL2, we are able to specifically deliver IL-2 to the tumor location, enhancing antigen-specific T-cell immune response and controlling tumor growth. Our approach represents a platform technology to specifically deliver proteins of interest to tumor loci.

  20. Human Embryonic Stem Cell-Derived Dopaminergic Neurons Reverse Functional Deficit in Parkinsonian Rats

    OpenAIRE

    Yang, Dali; Zhang, Zhi-jian; Oldenburg, Michael; Ayala, Melvin; Zhang, Su-Chun

    2007-01-01

    We show that human embryonic stem cell-derived dopaminergic neurons survived transplantation to the neurotoxin 6-hydroxydopamine-lesioned rat striatum and, in combination with the cells newly differentiated from their progenitors, contributed to locomotive function recovery at 5 months. The animal behavioral improvement was correlated with the dopamine neurons present in the graft. Although the donor cells contained forebrain and midbrain dopamine neurons, the dopamine neurons present in the ...

  1. A pure population of lung alveolar epithelial type II cells derived from human embryonic stem cells

    OpenAIRE

    Wang, Dachun; Haviland, David L.; Burns, Alan R.; Zsigmond, Eva; Wetsel, Rick A.

    2007-01-01

    Alveolar epithelial type II (ATII) cells are small, cuboidal cells that constitute ≈60% of the pulmonary alveolar epithelium. These cells are crucial for repair of the injured alveolus by differentiating into alveolar epithelial type I cells. ATII cells derived from human ES (hES) cells are a promising source of cells that could be used therapeutically to treat distal lung diseases. We have developed a reliable transfection and culture procedure, which facilitates, via genetic selection, the ...

  2. Microfluidic isolation of cancer-cell-derived microvesicles from hetergeneous extracellular shed vesicle populations

    OpenAIRE

    Santana, Steven M.; Antonyak, Marc A.; Cerione, Richard A.; Kirby, Brian J.

    2014-01-01

    Extracellular shed vesicles, including exosomes and microvesicles, are disseminated throughout the body and represent an important conduit of cell communication. Cancer-cell-derived microvesicles have potential as a cancer biomarker as they help shape the tumor microenvironment to promote the growth of the primary tumor and prime the metastatic niche. It is likely that, in cancer cell cultures, the two constituent extracellular shed vesicle subpopulations, observed in dynamic light scattering...

  3. Involvement of multiple myeloma cell-derived exosomes in osteoclast differentiation

    OpenAIRE

    Raimondi, L.; De Luca, A.; Amodio, N; Manno, M.; Raccosta, S; Taverna, S; Bellavia, D; Naselli, F; Fontana, S; Schillaci, O.; Giardino, R.; Fini, M.; Tassone, P; A. Santoro; De Leo, G

    2015-01-01

    Bone disease is the most frequent complication in multiple myeloma (MM) resulting in osteolytic lesions, bone pain, hypercalcemia and renal failure. In MM bone disease the perfect balance between bone-resorbing osteoclasts (OCs) and bone-forming osteoblasts (OBs) activity is lost in favour of OCs, thus resulting in skeletal disorders. Since exosomes have been described for their functional role in cancer progression, we here investigate whether MM cell-derived exosomes may be involved in OCs ...

  4. Automated Electrophysiological and Pharmacological Evaluation of Human Pluripotent Stem Cell-Derived Cardiomyocytes

    OpenAIRE

    Rajamohan, Divya; Kalra, Spandan; Duc Hoang, Minh; George, Vinoj; Staniforth, Andrew; Russell, Hugh; Yang, Xuebin; Denning, Chris

    2016-01-01

    Automated planar patch clamp systems are widely used in drug evaluation studies because of their ability to provide accurate, reliable, and reproducible data in a high-throughput manner. Typically, CHO and HEK tumorigenic cell lines overexpressing single ion channels are used since they can be harvested as high-density, homogenous, single-cell suspensions. While human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are physiologically more relevant, these cells are fragile, have compl...

  5. Stem Cell-Derived Exosomes: A Potential Alternative Therapeutic Agent in Orthopaedics

    OpenAIRE

    John Burke; Ravindra Kolhe; Monte Hunter; Carlos Isales; Mark Hamrick; Sadanand Fulzele

    2016-01-01

    Within the field of regenerative medicine, many have sought to use stem cells as a promising way to heal human tissue; however, in the past few years, exosomes (packaged vesicles released from cells) have shown more exciting promise. Specifically, stem cell-derived exosomes have demonstrated great ability to provide therapeutical benefits. Exosomal products can include miRNA, other genetic products, proteins, and various factors. They are released from cells in a paracrine fashion in order to...

  6. Chemokine receptors in cancer metastasis and cancer cell-derived chemokines in host immune response.

    Science.gov (United States)

    Koizumi, Keiichi; Hojo, Shozo; Akashi, Takuya; Yasumoto, Kazuo; Saiki, Ikuo

    2007-11-01

    The chemotactic cytokines called chemokines are a superfamily of small secreted cytokines that were initially characterized through their ability to prompt the migration of leukocytes. Attention has been focused on the chemokine receptors expressed on cancer cells because cancer cell migration and metastasis show similarities to leukocyte trafficking. CXC chemokine receptor 4 (CXCR4) was first investigated as a chemokine receptor that is associated with lung metastasis of breast cancers. Recently, CXCR4 was reported to be a key molecule in the formation of peritoneal carcinomatosis in gastric cancer. In the present review, we highlight current knowledge about the role of CXCR4 in cancer metastases. In contrast to chemokine receptors expressed on cancer cells, little is known about the roles of cancer cell-derived chemokines. Cancer tissue consists of both cancer cells and various stromal cells, and leukocytes that infiltrate into cancer are of particular importance in cancer progression. Although colorectal cancer invasion is regulated by the chemokine CCL9-induced infiltration of immature myeloid cells into cancer, high-level expression of cancer cell-derived chemokine CXCL16 increases infiltrating CD8(+) and CD4(+) T cells into cancer tissues, and correlates with a good prognosis. We discuss the conflicting biological effects of cancer cell-derived chemokines on cancer progression, using CCL9 and CXCL16 as examples. PMID:17894551

  7. A strategy to ensure safety of stem cell-derived retinal pigment epithelium cells.

    Science.gov (United States)

    Choudhary, Parul; Whiting, Paul John

    2016-09-02

    Cell replacement and regenerative therapy using embryonic stem cell-derived material holds promise for the treatment of several pathologies. However, the safety of this approach is of prime importance given the teratogenic potential of residual stem cells, if present in the differentiated cell product. Using the example of embryonic stem cell-derived retinal pigment epithelium (RPE) for the treatment of age-related macular degeneration, we present a novel strategy for ensuring the absence of stem cells in the RPE population. Based on an unbiased screening approach, we identify and validate the expression of CD59, a cell surface marker expressed on RPE but absent on stem cells. We further demonstrate that flow sorting on the basis of CD59 expression can effectively purify RPE and deplete stem cells, resulting in a population free from stem cell impurity. This purification helps to ensure removal of stem cells and hence increases the safety of cells that may be used for clinical transplantation. This strategy can potentially be applied to other pluripotent stem cell-derived material and help mitigate concerns of using such cells for therapy.

  8. Functional neuromuscular junctions formed by embryonic stem cell-derived motor neurons.

    Directory of Open Access Journals (Sweden)

    Joy A Umbach

    Full Text Available A key objective of stem cell biology is to create physiologically relevant cells suitable for modeling disease pathologies in vitro. Much progress towards this goal has been made in the area of motor neuron (MN disease through the development of methods to direct spinal MN formation from both embryonic and induced pluripotent stem cells. Previous studies have characterized these neurons with respect to their molecular and intrinsic functional properties. However, the synaptic activity of stem cell-derived MNs remains less well defined. In this study, we report the development of low-density co-culture conditions that encourage the formation of active neuromuscular synapses between stem cell-derived MNs and muscle cells in vitro. Fluorescence microscopy reveals the expression of numerous synaptic proteins at these contacts, while dual patch clamp recording detects both spontaneous and multi-quantal evoked synaptic responses similar to those observed in vivo. Together, these findings demonstrate that stem cell-derived MNs innervate muscle cells in a functionally relevant manner. This dual recording approach further offers a sensitive and quantitative assay platform to probe disorders of synaptic dysfunction associated with MN disease.

  9. Stem Cell-Derived Exosomes: A Potential Alternative Therapeutic Agent in Orthopaedics

    Directory of Open Access Journals (Sweden)

    John Burke

    2016-01-01

    Full Text Available Within the field of regenerative medicine, many have sought to use stem cells as a promising way to heal human tissue; however, in the past few years, exosomes (packaged vesicles released from cells have shown more exciting promise. Specifically, stem cell-derived exosomes have demonstrated great ability to provide therapeutical benefits. Exosomal products can include miRNA, other genetic products, proteins, and various factors. They are released from cells in a paracrine fashion in order to combat local cellular stress. Because of this, there are vast benefits that medicine can obtain from stem cell-derived exosomes. If exosomes could be extracted from stem cells in an efficient manner and packaged with particular regenerative products, then diseases such as rheumatoid arthritis, osteoarthritis, bone fractures, and other maladies could be treated with cell-free regenerative medicine via exosomes. Many advances must be made to get to this point, and the following review highlights the current advances of stem cell-derived exosomes with particular attention to regenerative medicine in orthopaedics.

  10. iPS-cell derived dendritic cells and macrophages for cancer therapy.

    Science.gov (United States)

    Senju, Satoru

    2016-08-01

    Antibody-based anti-cancer immunotherapy was recently recognized as one of the truly effective therapies for cancer patients. Antibodies against cell surface cancer antigens, such as CD20, and also those against immune-inhibitory molecules called "immune checkpoint blockers", such as CTLA4 or PD1, have emerged. Large-scale clinical trials have confirmed that, in some cases, antibody-based drugs are superior to conventional chemotherapeutic agents. These antibody-based drugs are now being manufactured employing a mass-production system by pharmaceutical companies. Anti-cancer therapy by immune cells, i.e. cell-based immunotherapy, is expected to be more effective than antibody therapy, because immune cells can recognize, infiltrate, and act in cancer tissues more directly than antibodies. In order to achieve cell-based anti-cancer immunotherapy, it is necessary to develop manufacturing systems for mass-production of immune cells. Our group has been studying immunotherapy with myeloid cells derived from ES cells or iPS cells. These pluripotent stem cells can be readily propagated under constant culture conditions, with expansion into a large quantity. We consider these stem cells to be the most suitable cellular source for mass-production of immune cells. This review introduces our studies on anti-cancer therapy with iPS cell-derived dendritic cells and iPS cell-derived macrophages. PMID:27599426

  11. Preparation of Attenuated Salmonella Typhimurium Strain Containing pCMV-IL-2-IRES-NK4 Plasmid and Its Stability%含 pCMV-IL-2-IRES-NK4质粒减毒沙门菌的制备及其稳定性研究

    Institute of Scientific and Technical Information of China (English)

    杨志华; 哈小琴; 张尚弟; 冯强生; 薛荣利; 杨淑娟; 赵勇; 杨迎桂

    2014-01-01

    目的:制备含 pCMV-IL-2-IRES-NK4质粒减毒沙门菌 TPIN 并检测其稳定性。方法将 pCMV-IL-2-IRES-NK4质粒转进减毒沙门菌 Ty21a 感受态,制备成重组的减毒沙门菌菌株 TPIN;将 TPIN 在含有氨苄西林(+)和氨苄西林(-)的 LB 培养板传代培养至第10代、20代、30代和40代时用灭菌的牙签分别挑取含有氨苄西林(+)和氨苄西林(-)LB 培养基上的单克隆菌落,质粒提取、PCR 扩增酶切鉴定;TPIN 转染 HepG2细胞后采用 RT-PCR 检测IL-2和 NK4基因表达,ELISA 法检测细胞培养上清 IL-2和 NK4蛋白。结果构建菌株经提取质粒、酶切、PCR 扩增获得目的基因 IL-2、NK4特异条带;在氨苄西林(+)和氨苄西林(-)LB 培养板传代培养40代的 TPIN 菌株均可扩增并双酶切出目的基因 IL-2及 NK4;TPIN 体外转染 HepG2细胞后,IL-2及 NK4表达水平均显著升高。结论重组携带IL-2/ NK4双基因的减毒沙门菌菌株 TPIN 可稳定遗传,不受无选择性压力的影响而丢失质粒,且在体外 IL-2、NK4基因及蛋白可以稳定高效表达。%Objective To prepare an attenuated Salmonella typhimurium DNA vaccine containing pCMV-IL-2-IRES-NK4 plasmid (TPIN) and to detect its stability. Methods The pCMV-IL-2-IRES-NK4 plasmid was transformed into an attenuated Salmonella typhimurium Ty21a competence, and then a recombinant attenuated Salmonella typhimuri-um strain TPIN was prepared; the recombinant strain was cultivated in 40 generations on LB nutritional medium plate with or without Ampicillin, and monoclonal bacterial colony was selected at subculture 10th , 20th , 30th and 40th generation from Ampicillin ( + ) and no Ampicillin ( - ) plates respectively, and then the plasmids were extracted and identified by PCR amplification and enzyme digestion; TPIN was transfected into HepG2 cells, then expressions of the IL-2 / NK4 genes were determined by reverse transcription polymerase chain reaction (RT-PCR), and expressions of IL-2 / NK4

  12. Cardiomyocyte MEA data analysis (CardioMDA--a novel field potential data analysis software for pluripotent stem cell derived cardiomyocytes.

    Directory of Open Access Journals (Sweden)

    Paruthi Pradhapan

    Full Text Available Cardiac safety pharmacology requires in-vitro testing of all drug candidates before clinical trials in order to ensure they are screened for cardio-toxic effects which may result in severe arrhythmias. Micro-electrode arrays (MEA serve as a complement to current in-vitro methods for drug safety testing. However, MEA recordings produce huge volumes of data and manual analysis forms a bottleneck for high-throughput screening. To overcome this issue, we have developed an offline, semi-automatic data analysis software, 'Cardiomyocyte MEA Data Analysis (CardioMDA', equipped with correlation analysis and ensemble averaging techniques to improve the accuracy, reliability and throughput rate of analysing human pluripotent stem cell derived cardiomyocyte (CM field potentials. With the program, true field potential and arrhythmogenic complexes can be distinguished from one another. The averaged field potential complexes, analysed using our software to determine the field potential duration, were compared with the analogous values obtained from manual analysis. The reliability of the correlation analysis algorithm, evaluated using various arrhythmogenic and morphology changing signals, revealed a mean sensitivity and specificity of 99.27% and 94.49% respectively, in determining true field potential complexes. The field potential duration of the averaged waveforms corresponded well to the manually analysed data, thus demonstrating the reliability of the software. The software has also the capability to create overlay plots for signals recorded under different drug concentrations in order to visualize and compare the magnitude of response on different ion channels as a result of drug treatment. Our novel field potential analysis platform will facilitate the analysis of CM MEA signals in semi-automated way and provide a reliable means of efficient and swift analysis for cardiomyocyte drug or disease model studies.

  13. Niche-dependent development of functional neuronal networks from embryonic stem cell-derived neural populations

    Directory of Open Access Journals (Sweden)

    Siebler Mario

    2009-08-01

    Full Text Available Abstract Background The present work was performed to investigate the ability of two different embryonic stem (ES cell-derived neural precursor populations to generate functional neuronal networks in vitro. The first ES cell-derived neural precursor population was cultivated as free-floating neural aggregates which are known to form a developmental niche comprising different types of neural cells, including neural precursor cells (NPCs, progenitor cells and even further matured cells. This niche provides by itself a variety of different growth factors and extracellular matrix proteins that influence the proliferation and differentiation of neural precursor and progenitor cells. The second population was cultivated adherently in monolayer cultures to control most stringently the extracellular environment. This population comprises highly homogeneous NPCs which are supposed to represent an attractive way to provide well-defined neuronal progeny. However, the ability of these different ES cell-derived immature neural cell populations to generate functional neuronal networks has not been assessed so far. Results While both precursor populations were shown to differentiate into sufficient quantities of mature NeuN+ neurons that also express GABA or vesicular-glutamate-transporter-2 (vGlut2, only aggregate-derived neuronal populations exhibited a synchronously oscillating network activity 2–4 weeks after initiating the differentiation as detected by the microelectrode array technology. Neurons derived from homogeneous NPCs within monolayer cultures did merely show uncorrelated spiking activity even when differentiated for up to 12 weeks. We demonstrated that these neurons exhibited sparsely ramified neurites and an embryonic vGlut2 distribution suggesting an inhibited terminal neuronal maturation. In comparison, neurons derived from heterogeneous populations within neural aggregates appeared as fully mature with a dense neurite network and punctuated

  14. CD4、CD8和IL-2在海洛因戒断、复吸大鼠胃窦的表达%Expression of CD4, CD8 and IL-2 in the gastric antrum of rats during heroin abstinence and relapse

    Institute of Scientific and Technical Information of China (English)

    夏白娟; 梁文妹; 李一欣; 洪艳; 韩晶; 胡赟

    2013-01-01

    AIM:To detect the expression of CD4,CD8 and interleukin-2 (IL-2) in the gastric antrum of rats during heroin abstinence and relapse.METHODS:Thirty-five male adult SD rats were randomly divided into three groups:normal control group (NCG),saline control group (SCG) and experiment group (EG).The experiment group was divided into three subgroups:heroin abstinence subgroup (HAS),methadone detoxication subgroup (MDS) and heroin relapse subgroup (HRS).Immunohistochemistry was performed to detect the expression of CD4,CD8 and IL-2 in the gastric antrum of rats of different groups.RESULTS:Compared to the NCG,the staining intensity of CD4 and IL-2 (CD4:135.314 ± 3.595 vs 154.889 ± 3.238,155.455 ± 3.176; IL-2:131.082 ± 4.769 vs 150.233 ± 4.241,150.199 ± 4.007; all P <0.05) and the numbers of CD4-and IL-2-positive cells (CD4:15.944 ± 1.661 vs 8.500 ± 1.150,10.278 ± 1.074; IL-2:21.824 ± 2.556 vs 13.118 ± 3.180,8.529 ± 1.179; all P < 0.05) in the gastric antrum decreased,and the staining intensity of CD8 038.239 ± 4.047 vs 123.717 ± 4.622,125.498 ± 3.056,both P < 0.05) and the number of CD8-positive cells (18.611 ± 2.477 vs 26.167 ± 2.203,26.778 ± 1.592,both P < 0.05) significantly increased in the HAS and HRS.Compared to the NCG and SCG,there were no obvious changes in the staining intensity of CD4,CD8 and IL-2 and the numbers of CD4-,CD8-and IL-2-positive ceils in the MDS (all P > 0.05).CONCLUSION:The expression of CD4 and IL-2 decreases and that of CD8 increases in the gastric antrum of rats during heroin abstinence and relapse,which suggests that the antrum is damaged during heroin abstinence and relapse.In the MDG,the expression of CD4,CD8 and IL-2 in the MDS is close to that in the NCG and SCG,suggesting that the structure and function of the antrum could be restored after detoxication with methadone.%目的:探讨海洛因戒断、复吸对大鼠胃窦内CD4、CD8和白介素2(interleukin-2,IL-2)表达的影响.方法:正常♂SD大鼠35只,

  15. Xenogeneic Graft-versus-Host-Disease in NOD-scid IL-2Rγnull Mice Display a T-Effector Memory Phenotype

    OpenAIRE

    Niwa Ali; Barry Flutter; Robert Sanchez Rodriguez; Ehsan Sharif-Paghaleh; Barber, Linda D.; Giovanna Lombardi; Nestle, Frank O

    2012-01-01

    The occurrence of Graft-versus-Host Disease (GvHD) is a prevalent and potentially lethal complication that develops following hematopoietic stem cell transplantation. Humanized mouse models of xenogeneic-GvHD based upon immunodeficient strains injected with human peripheral blood mononuclear cells (PBMC; "Hu-PBMC mice") are important tools to study human immune function in vivo. The recent introduction of targeted deletions at the interleukin-2 common gamma chain (IL-2Rγ(null)), notably the N...

  16. Spontaneous Post-Transplant Disorders in NOD.Cg- Prkdcscid Il2rgtm1Sug/JicTac (NOG) Mice Engrafted with Patient-Derived Metastatic Melanomas

    OpenAIRE

    Enrico Radaelli; Els Hermans; Lorna Omodho; Annick Francis; Sara Vander Borght; Jean-Christophe Marine; Joost van den Oord; Frédéric Amant

    2015-01-01

    Patient-derived tumor xenograft (PDTX) approach is nowadays considered a reliable preclinical model to study in vivo cancer biology and therapeutic response. NOD scid and Il2rg-deficient mice represent the “gold standard” host for the generation of PDTXs. Compared to other immunocompromised murine lines, these mice offers several advantages including higher engraftment rate, longer lifespan and improved morphological and molecular preservation of patient-derived neoplasms. Here we describe a ...

  17. Human acute myelogenous leukemia stem cells are rare and heterogeneous when assayed in NOD/SCID/IL2Rγc-deficient mice

    OpenAIRE

    Sarry, Jean-Emmanuel; Murphy, Kathleen; Perry, Robin; Sanchez, Patricia V.; Secreto, Anthony; Keefer, Cathy; Swider, Cezary R.; Strzelecki, Anne-Claire; Cavelier, Cindy; Récher, Christian; Mansat-De Mas, Véronique; Delabesse, Eric; Danet-Desnoyers, G; Carroll, Martin

    2010-01-01

    Human leukemic stem cells, like other cancer stem cells, are hypothesized to be rare, capable of incomplete differentiation, and restricted to a phenotype associated with early hematopoietic progenitors or stem cells. However, recent work in other types of tumors has challenged the cancer stem cell model. Using a robust model of xenotransplantation based on NOD/SCID/IL2Rγc-deficient mice, we confirmed that human leukemic stem cells, functionally defined by us as SCID leukemia-initiating cells...

  18. Mucosal Delivery of Murine Interleukin-2 (IL-2) and IL-6 by Recombinant Strains of Lactococcus lactis Coexpressing Antigen and Cytokine

    OpenAIRE

    Steidler, Lothar; Robinson, K.; Chamberlain, L.; SCHOFIELD, KM; Remaut, Erik; LE PAGE, RWF; Wells, JM

    1998-01-01

    Lactococcus lactis is a nonpathogenic and noncolonizing bacterium which is being developed as a vaccine delivery vehicle for immunization by mucosal routes. To determine whether lactococci can also deliver cytokines to the immune system, we have constructed novel constitutive expression strains of L. lactis which accumulate a test antigen, tetanus toxin fragment C (TTFC), within the cytoplasmic compartment and also secrete either murine interleukin-2 (IL-2) or IL-6. When mice were immunized i...

  19. Clinical observation of serum IL-18, IL-10 and sIL-2R levels in patients with chronic hepatitis C pre- and post antiviral treatment

    Institute of Scientific and Technical Information of China (English)

    贾红宇; 杜杰; 朱思和; 马英骥; 蔡华枫

    2003-01-01

    Objective To discuss the roles of serum interleukin-18 (IL-18), interleukin-10 (IL-10) and soluble interleukin-2R (sIL-2R) in the pathogenesis of chronic hepatitis C and to observe the effects of interferon (IFN) on the above- mentioned serum cytokines. Methods The levels of above- mentioned cytokines were detected in 10 healthy individuals, 24 asymptomatic hepatitis virus C (HCV) carriers and 27 patients with chronic hepatitis C ( before and after IFN treatment) using enzyme linked immunosorbent assay (ELISA). Results The levels of the cytokines in patients with chronic hepatitis C are higher than in healthy people (P<0.05) and in asymptomatic HCV carriers(P<0.05). The values of the cytokines show a significant positive correlation to ALT (P<0.05). Levels of tested cytokines decreased observably after IFN treatment (P<0.05). The grades of the serum levels for sIL-2R and IL-10 before IFN treatment (from high to low) were categorized accordingly: non-response group> partial- response group >complete- response group (P<0.05). Conclusions The tested cytokines co-participate in the pathogenesis of chronic hepatitis C, and can be used to evaluate the effect of IFN on the immune state of organisms. Furthermore, sIL-2R and IL-10 are important for predicting the anti-viral efficacy of IFN.

  20. Protective role of TNF-α, IL-10 and IL-2 in mice infected with the Oshima strain of Tick-borne encephalitis virus

    Science.gov (United States)

    Tun, Mya Myat Ngwe; Aoki, Kotaro; Senba, Masachika; Buerano, Corazon C.; Shirai, Kenji; Suzuki, Ryuji; Morita, Kouichi; Hayasaka, Daisuke

    2014-01-01

    Tick-borne encephalitis virus (TBEV) causes acute central nervous system disease. Here, we investigated the roles of the TNF-α, IL-10 and other cytokines in appropriate KO mice following infection with Oshima and Sofjin strains of TBEV. Following infection with the Oshima strain, mortality rates were significantly increased in TNF-α KO and IL-10 KO mice compared with wild type (WT) mice. These results suggested that TNF-α and IL-10 play protective roles against fatal infection due to Oshima strain infection. However, viral loads and proinflammatory cytokine levels in the brain of TNF-α KO andIL-10 KO mice were not significantly different compared with those of WT mice. On the other hand, all WT, TNF-α KO and IL-10 KO mice died following infection with Sofjin strain. Interestingly, Sofjin-infected mice did not exhibit an up-regulated mRNA level of IL-2 in the spleen in all groups of mice, whereas Oshima-infected mice showed significantly increased level of IL-2 compared with mock-infected mice. From these results, we suggest that TNF-α, IL-10 and IL-2 are key factors for disease remission from fatal encephalitis due to infection with Oshima strain of TBEV. PMID:24938868

  1. The Rag2⁻Il2rb⁻Dmd⁻ mouse: a novel dystrophic and immunodeficient model to assess innovating therapeutic strategies for muscular dystrophies.

    Science.gov (United States)

    Vallese, Denis; Negroni, Elisa; Duguez, Stéphanie; Ferry, Arnaud; Trollet, Capucine; Aamiri, Ahmed; Vosshenrich, Christian A J; Füchtbauer, Ernst-Martin; Di Santo, James P; Vitiello, Libero; Butler-Browne, Gillian; Mouly, Vincent

    2013-10-01

    The development of innovative therapeutic strategies for muscular dystrophies, particularly cell-based approaches, is still a developing field. Although positive results have been obtained in animal models, they have rarely been confirmed in patients and resulted in very limited clinical improvements, suggesting some specificity in humans. These findings emphasized the need for an appropriate animal model (i.e., immunodeficient and dystrophic) to investigate in vivo the behavior of transplanted human myogenic stem cells. We report a new model, the Rag2(-)Il2rb(-)Dmd(-) mouse, which lacks T, B, and NK cells, and also carries a mutant Dmd allele that prevents the production of any dystrophin isoform. The dystrophic features of this new model are comparable with those of the classically used mdx mouse, but with the total absence of any revertant dystrophin positive fiber. We show that Rag2(-)Il2rb(-)Dmd(-) mice allow long-term xenografts of human myogenic cells. Altogether, our findings indicate that the Rag2(-)Il2rb(-)Dmd(-) mouse represents an ideal model to gain further insights into the behavior of human myogenic stem cells in a dystrophic context, and can be used to assess innovative therapeutic strategies for muscular dystrophies. PMID:23975040

  2. 实验性自身免疫性卵巢功能早衰中IL-2和IFN-γ的检测

    Institute of Scientific and Technical Information of China (English)

    杨桂华; 柯丹

    2007-01-01

    目的:探讨Th1型细胞因子IL-2和IFN-γ在自身免疫性卵巢功能早衰(POF)中的表达。方法:分别用猪卵透明带(PZP)抗原和PBS缓冲液注射免疫Balb/c小鼠,共免疫3次,每2W免疫1次,经过3次免疫后,观察各组卵巢组织病理形态,并测定各组IL-2和IFN-γ水平。结果:PZP免疫小鼠卵巢组织形态明显改变,且Th1性细胞因子IL-2和IFN-γ表达均明显高于对照组。结论:自身免疫性卵巢功能早衰与T细胞介导的Th1型免疫应答有关。

  3. 实验性自身免疫性卵巢功能早衰中IL-2和IFN-γ的检测

    Institute of Scientific and Technical Information of China (English)

    杨桂华; 柯丹

    2007-01-01

    目的:探讨Th1型细胞因子IL-2和IFN-γ在自身免疫性卵巢功能早衰(POF)中的表达。方法:分别用猪卵透明带(PZP)抗原和PBS缓冲液注射免疫Balb/c小鼠,共免疫3次,每2W免疫1次,经过3次免疫后,观察各组卵巢组织病理形态,并测定各组IL-2和IFN-γ水平。结果:PZP免疫小鼠卵巢组织形态明显改变,且Th14性细胞因子IL-2和IFN-γ表达均明显高于对照组。结论:自身免疫性卵巢功能早衰与T细胞介导的Th1型免疫应答有关。

  4. N-butylidenephthalide attenuates Alzheimer's disease-like cytopathy in Down syndrome induced pluripotent stem cell-derived neurons.

    Science.gov (United States)

    Chang, Chia-Yu; Chen, Sheng-Mei; Lu, Huai-En; Lai, Syu-Ming; Lai, Ping-Shan; Shen, Po-Wen; Chen, Pei-Ying; Shen, Ching-I; Harn, Horng-Jyh; Lin, Shinn-Zong; Hwang, Shiaw-Min; Su, Hong-Lin

    2015-03-04

    Down syndrome (DS) patients with early-onset dementia share similar neurodegenerative features with Alzheimer's disease (AD). To recapitulate the AD cell model, DS induced pluripotent stem cells (DS-iPSCs), reprogrammed from mesenchymal stem cells in amniotic fluid, were directed toward a neuronal lineage. Neuroepithelial precursor cells with high purity and forebrain characteristics were robustly generated on day 10 (D10) of differentiation. Accumulated amyloid deposits, Tau protein hyperphosphorylation and Tau intracellular redistribution emerged rapidly in DS neurons within 45 days but not in normal embryonic stem cell-derived neurons. N-butylidenephthalide (Bdph), a major phthalide ingredient of Angelica sinensis, was emulsified by pluronic F127 to reduce its cellular toxicity and promote canonical Wnt signaling. Interestingly, we found that F127-Bdph showed significant therapeutic effects in reducing secreted Aβ40 deposits, the total Tau level and the hyperphosphorylated status of Tau in DS neurons. Taken together, DS-iPSC derived neural cells can serve as an ideal cellular model of DS and AD and have potential for high-throughput screening of candidate drugs. We also suggest that Bdph may benefit DS or AD treatment by scavenging Aβ aggregates and neurofibrillary tangles.

  5. Effect of stromal-cell-derived factor 1 on stem-cell homing and tissue regeneration in ischaemic cardiomyopathy

    Science.gov (United States)

    Askari, Arman T.; Unzek, Samuel; Popovic, Zoran B.; Goldman, Corey K.; Forudi, Farhad; Kiedrowski, Matthew; Rovner, Aleksandr; Ellis, Stephen G.; Thomas, James D.; DiCorleto, Paul E.; Topol, Eric J.; Penn, Marc S.

    2003-01-01

    BACKGROUND: Myocardial regeneration via stem-cell mobilisation at the time of myocardial infarction is known to occur, although the mechanism for stem-cell homing to infarcted tissue subsequently and whether this approach can be used for treatment of ischaemic cardiomyopathy are unknown. We investigated these issues in a Lewis rat model (ligation of the left anterior descending artery) of ischaemic cardiomyopathy. METHODS: We studied the effects of stem-cell mobilisation by use of granulocyte colony-stimulating factor (filgrastim) with or without transplantation of syngeneic cells. Shortening fraction and myocardial strain by tissue doppler imaging were quantified by echocardiography. FINDINGS: Stem-cell mobilisation with filgrastim alone did not lead to engraftment of bone-marrow-derived cells. Stromal-cell-derived factor 1 (SDF-1), required for stem-cell homing to bone marrow, was upregulated immediately after myocardial infarction and downregulated within 7 days. 8 weeks after myocardial infarction, transplantation into the peri-infarct zone of syngeneic cardiac fibroblasts stably transfected to express SDF-1 induced homing of CD117-positive stem cells to injured myocardium after filgrastim administration (control vs SDF-1-expressing cardiac fibroblasts mean 7.2 [SD 3.4] vs 33.2 [6.0] cells/mm2, n=4 per group, pmyocardial tissue must be initiated within days of myocardial infarction unless signalling for stem-cell homing is re-established.

  6. Lessons from the heart: mirroring electrophysiological characteristics during cardiac development to in vitro differentiation of stem cell derived cardiomyocytes.

    Science.gov (United States)

    van den Heuvel, Nikki H L; van Veen, Toon A B; Lim, Bing; Jonsson, Malin K B

    2014-02-01

    The ability of human pluripotent stem cells (hPSCs) to differentiate into any cell type of the three germ layers makes them a very promising cell source for multiple purposes, including regenerative medicine, drug discovery, and as a model to study disease mechanisms and progression. One of the first specialized cell types to be generated from hPSC was cardiomyocytes (CM), and differentiation protocols have evolved over the years and now allow for robust and large-scale production of hPSC-CM. Still, scientists are struggling to achieve the same, mainly ventricular, phenotype of the hPSC-CM in vitro as their adult counterpart in vivo. In vitro generated cardiomyocytes are generally described as fetal-like rather than adult. In this review, we compare the in vivo development of cardiomyocytes to the in vitro differentiation of hPSC into CM with focus on electrophysiology, structure and contractility. Furthermore, known epigenetic changes underlying the differences between adult human CM and CM differentiated from pluripotent stem cells are described. This should provide the reader with an extensive overview of the current status of human stem cell-derived cardiomyocyte phenotype and function. Additionally, the reader will gain insight into the underlying signaling pathways and mechanisms responsible for cardiomyocyte development.

  7. Effect of stromal-cell-derived factor 1 on stem-cell homing and tissue regeneration in ischaemic cardiomyopathy

    Science.gov (United States)

    Askari, Arman T.; Unzek, Samuel; Popovic, Zoran B.; Goldman, Corey K.; Forudi, Farhad; Kiedrowski, Matthew; Rovner, Aleksandr; Ellis, Stephen G.; Thomas, James D.; DiCorleto, Paul E.; Topol, Eric J.; Penn, Marc S.

    2003-01-01

    BACKGROUND: Myocardial regeneration via stem-cell mobilisation at the time of myocardial infarction is known to occur, although the mechanism for stem-cell homing to infarcted tissue subsequently and whether this approach can be used for treatment of ischaemic cardiomyopathy are unknown. We investigated these issues in a Lewis rat model (ligation of the left anterior descending artery) of ischaemic cardiomyopathy. METHODS: We studied the effects of stem-cell mobilisation by use of granulocyte colony-stimulating factor (filgrastim) with or without transplantation of syngeneic cells. Shortening fraction and myocardial strain by tissue doppler imaging were quantified by echocardiography. FINDINGS: Stem-cell mobilisation with filgrastim alone did not lead to engraftment of bone-marrow-derived cells. Stromal-cell-derived factor 1 (SDF-1), required for stem-cell homing to bone marrow, was upregulated immediately after myocardial infarction and downregulated within 7 days. 8 weeks after myocardial infarction, transplantation into the peri-infarct zone of syngeneic cardiac fibroblasts stably transfected to express SDF-1 induced homing of CD117-positive stem cells to injured myocardium after filgrastim administration (control vs SDF-1-expressing cardiac fibroblasts mean 7.2 [SD 3.4] vs 33.2 [6.0] cells/mm2, n=4 per group, pfunction (shortening fraction 9.2 [4.9] vs 17.2 [4.2]%, n=8 per group, pmyocardial tissue must be initiated within days of myocardial infarction unless signalling for stem-cell homing is re-established.

  8. TNF-alpha associated with extracellular matrix fibronectin provides a stop signal for chemotactically migrating T cells.

    Science.gov (United States)

    Franitza, S; Hershkoviz, R; Kam, N; Lichtenstein, N; Vaday, G G; Alon, R; Lider, O

    2000-09-01

    The migration of T cells into extravascular sites of inflammation is regulated by information derived from the molecular structure of the invaded tissue and from chemokine and cytokine gradients in the context of the extracellular matrix (ECM). Although recent studies have highlighted the role of particular chemoattractants in leukocyte migration, to date little is known about how specific combinations of contextual signals control the migration of leukocytes and their localization at sites of inflammation. Here we studied the interplay between a pleiotropic cytokine, TNF-alpha, and two prototypic chemoattractants, RANTES and stromal cell-derived factor-1alpha (SDF-1alpha), on human CD45RO+ T cells migrating within an ECM-like context. For this purpose, we used a newly constructed three-dimensional gel system designed to follow, in real time, the migration of individual leukocytes along chemotactic gradients in vitro. We found that TNF-alpha, which binds the ECM protein fibronectin and lacks adhesion- and migration-promoting effects of its own, can act as a proadhesive cytokine on T cells exposed to RANTES and SDF-1alpha. Furthermore, fibronectin-complexed TNF-alpha provided anchorage signals to the T cells as they moved directionally along chemoattractive gradients. This effect of TNF-alpha required an intact TNF-alpha receptor II subtype on the migrating T cells. The anchoring effect of TNF-alpha appears to be specific; IL-2, an integrin-activating proadhesive cytokine, does not transmit stoppage signals to T cell migration induced by RANTES. Thus, TNF-alpha present in the ECM at sites of inflammation may function to anchor T cells recruited to these sites by chemotactic signals. PMID:10946305

  9. Effect of Long-term Inhalation of Glucocorticoids on the Level of Leptin, IL-13 and IL-2 in Bronchial Asthmatic Children%长期吸入糖皮质激素对支气管哮喘患者血清leptin、IL-13和IL-2水平的影响

    Institute of Scientific and Technical Information of China (English)

    潘炯伟

    2011-01-01

    目的:观察支气管哮喘患者血清leptin、IL-13和IL-2水平的变化并探讨长期吸入糖皮质激素后对支气管哮喘患者血清leptin、IL-13和IL-2的影响.方法:4采用酶联免疫吸附法(ELISA),分别检测70例支气管哮喘患者采用吸入糖皮质激素治疗前、治疗3、6、12个月后及60例对照者血清IL-13和IL-2水平;采用放射免疫分析血清leptin浓度.结果:支气管哮喘患者血清leptin、IL-13和IL-2水平显著高于正常对照组.吸入糖皮质激素治疗3个月后支气管哮喘患者血清leptin、IL-13和IL-2浓度与治疗前比较下降(P均<0.05),治疗6个月和12个月后显著低于治疗前(P均<0.01),且治疗12个月后各个炎症指标与对照组相比无显著性差异(P>0.05).结论:长期吸入糖皮质激素是治疗支气管哮喘的重要手段,其治疗作用与下调血清leptin、IL-13和IL-2水平有关.%Objective To determine the effect of long-term inhalation of glucocorticoids on the level of leptin, IL-13, and H-2 in bronchial asthmatic patient. Methods End/me-linked immunosorbent assay ( ELISA ) was used to detect the serum IL-13 and IL-2 level in 60 healthy persons ( normal control group ) and 70 bronchial asthma patients untreated and 3, 6, 12 months pest-treatment, meanwhile leptin was determined by radio immunoassay. Results The serum levels of leptin, IL-13, and IL-2 in were significantly increased in patient with bronchial asthma compared with that in the normal control group. The serum levels of leptin, IL-13, and IL 2 in children with asthma were clem'eased gradually after inhaling gineccortieoids for 3 months ( P < 0.05 ). The treatment of inhaled glucocortieoids for 6 and 12 months can attenuate the elevation of leptin, IL-13, and IL-2 compared with that before the treatment. Conclusion Long-term inhaled glucoeortieoid is an important means for asthma, and the effects are related to the decrease of level of leptin, IL-13, and IL-2.

  10. Association between negative life events and the level changes of serum cytokine (IL-2, IL-6 and TNF-α) in patients with first-episode depression%负性生活事件与首发抑郁症患者血清IL-2、IL-6及TNF-α水平的关联分析

    Institute of Scientific and Technical Information of China (English)

    陈帅; 马玉璐; 刘素芳

    2015-01-01

    目的:探讨负性生活事件对首发抑郁症患者血清细胞因子il-2、il-6及tnf-α水平的影响。方法采用酶联免疫吸附法(elisa)对有负性生活事件诱因的抑郁症患者30例、无诱因的抑郁症患者30例和30名正常人的血清白介素2(il-2)、白介素6(il-6)和肿瘤坏死因子-α(tnf-α)水平进行检测。结果 il-2、il-6及tnf-α水平在3组间存在显著性差异,血清il-2水平有诱因组明显低于无诱因组和对照组(P0.05);患者组血清il-6和tnf-α水平均明显高于对照组(P0.05)。血清il-2水平与负性生活事件刺激量总分呈负相关,与HaMd总分呈正相关。结论负性生活事件对抑郁症患者的细胞免疫激活系统有抑制作用,il-2可能是起重要的作用。%Objective: to explore the effects of negative life events on serum cytokine levels in patients with first-episode depression.Methods:the levels of serum interleukin-2(il-2), interleukin-6 (il-6)and tumor necrosis factor-α(tnf-α)were measured in 30 patients with negative life events induced depression, 30 patients with non-life events induced depression and 30 normal controls by enzyme linked immunosorbent assay (elisa). Results:the levels of serum il-2 was lower in negative life event group than non-life event group and control group. However, the difference in il-2 levels were not obvious between non-life event group and control group (P>0.05). the levels of serum il-6 and tnf-αwere highter in patients than control group, however we fail to found obvious difference between negative life event group and non-life event group. We found a negative correlation with negative life events scores and a positive correlation with HaMd scores in the levels of il-2 for negative life event group. Conclusion:negative life events have suppressing action on cellular immunity activating system and il-2 may paly a vital role.

  11. Lung epithelial cell-derived extracellular vesicles activate macrophage-mediated inflammatory responses via ROCK1 pathway.

    Science.gov (United States)

    Moon, H-G; Cao, Y; Yang, J; Lee, J H; Choi, H S; Jin, Y

    2015-01-01

    Despite decades of research, the pathogenesis of acute respiratory distress syndrome (ARDS) remains poorly understood, thus impeding the development of effective treatment. Diffuse alveolar damage (DAD) and lung epithelial cell death are prominent features of ARDS. Lung epithelial cells are the first line of defense after inhaled stimuli, such as in the case of hyperoxia. We hypothesized that lung epithelial cells release 'messenger' or signaling molecules to adjacent or distant macrophages, thereby initiating or propagating inflammatory responses after noxious insult. We found that, after hyperoxia, a large amount of extracellular vesicles (EVs) were generated and released into bronchoalveolar lavage fluid (BALF). These hyperoxia-induced EVs were mainly derived from live lung epithelial cells as the result of hyperoxia-associated endoplasmic reticulum (ER) stress. These EVs were remarkably different from epithelial 'apoptotic bodies', as reflected by the significantly smaller size and differentially expressed protein markers. These EVs fall mainly in the size range of the exosomes and smaller microvesicles (MVs) (50-120 nm). The commonly featured protein markers of apoptotic bodies were not found in these EVs. Treating alveolar macrophages with hyperoxia-induced, epithelial cell-derived EVs led to an increased secretion of pro-inflammatory cytokines and macrophage inflammatory protein 2 (MIP-2). Robustly increased macrophage and neutrophil influx was found in the lung tissue of the mice intranasally treated with hyperoxia-induced EVs. It was determined that EV-encapsulated caspase-3 was largely responsible for the alveolar macrophage activation via the ROCK1 pathway. Caspase-3-deficient EVs induced less cytokine/MIP-2 release, reduced cell counts in BALF, less neutrophil infiltration and less inflammation in lung parenchyma, both in vitro and in vivo. Furthermore, the serum circulating EVs were increased and mainly derived from lung epithelial cells after

  12. Neuronal-like cell differentiation of non-adherent bone marrow cell-derived mesenchymal stem cells

    OpenAIRE

    Wu, Yuxin; Zhang, Jinghan; Ben, Xiaoming

    2013-01-01

    Non-adherent bone marrow cell-derived mesenchymal stem cells from C57BL/6J mice were separated and cultured using the “pour-off” method. Non-adherent bone marrow cell-derived mesenchymal stem cells developed colony-forming unit-fibroblasts, and could be expanded by supplementation with epidermal growth factor. Immunocytochemistry showed that the non-adherent bone marrow cell-derived mesenchymal stem cells exposed to basic fibroblast growth factor/epidermal growth factor/nerve growth factor ex...

  13. Effect of alpha 2b interferon on inducement of mIL-2R and treatment of HCV in PBMC from patients with chronic viral hepatitis C

    Institute of Scientific and Technical Information of China (English)

    Jian Wang; Gui-Ju Xiang; Bing-Xiang Liu

    2003-01-01

    AIM: To study the level of membrane interleukin-2 receptor (mIL-2R) on surface of peripheral blood mononuclear cells (PBMC) and the therapeutic efficacy of alpha 2b interferon on the treatment of HCV-RNA in PBMC of patients with chronic hepatitis C and to compare the negative rates of HCV-RNA in PBMC, HCV-RNA and anti-HCV in serum.METHODS: Before and after treatment of alpha 2b interferon, the level of mIL-2R of patients with chronic hepatitis C was detected by biotin-streptavidin (BSA). The therapeutic group (26 cases) was treated with alpha 2b interferon (3 MU/d) and control therapeutic group (22 cases)was treated with routine drugs (VitC, aspartic acid). The total course of treatment with alpha 2b interferon and routine drug was six months and per course of the treatment was three months. The levels of HCV-RNA in PBMC, HCV-RNA and anti-HCV in serum were detected before and after a course of the treatment.RESULTS: Before and after treatment of alpha 2b interferon and routine drugs, the levels of mIL-2R in silence stage were (3.44±0.77)% and (2.95±0.72)%, the levels of mIL-2R in inducement stage were (33.62±3.95)% and (30.04±3.73)%. There was a significant difference between two groups (P<0.01-P<0.05). After treatment of alpha 2b interferon with 3 MU/d for two courses of the treatment,the total negative rates of HCV-RNA in the PBMC and HCVRNA, anti-HCV in serum were 42.31% (11/26), 57.69%(15/26), 65.38%(17/26) respectively. After the treatment of routine drug, the negative rates of HCV-RNA in PBMC and HCV-RNA, anti-HCV in serum were 13.64% (3/22),22.73% (5/22), 27.27% (6/22) respectively. There was high significant difference in the group treated with alpha 2b interferon and the group treated with routine drugs (P<0.01-P<0.05).CONCLUSION: The mIL-2R can be induced by alpha 2b interferon during the treatment. The alpha 2b interferon has a definite effect on the treatment of HCV-RNA in PBMC.The curative effect of alpha 2b interferon is better than that

  14. Interleukin-2 Enhances the Regulatory Functions of CD4(+)T Cell-Derived CD4(-)CD8(-) Double Negative T Cells.

    Science.gov (United States)

    Cong, Min; Liu, Tianhui; Tian, Dan; Guo, Hongbo; Wang, Ping; Liu, Kai; Lin, Jun; Tian, Yue; Shi, Wen; You, Hong; Jia, Jidong; Zhang, Dong

    2016-08-01

    CD4(+) T cells can be converted to CD4(-)CD8(-) double negative T cells (DN T cells) under appropriate conditions, and IL-2 enhanced the conversion. Here, we investigated the effect of IL-2 on the proliferation and function of converted DN T cells in vitro and in vivo. DN T cells were hyporesponsive when restimulated by mature dendritic cells (mDCs), IL-2 completely restored their responsiveness in vitro. In addition, IL-2 increased the resistance of DN T cells to apoptosis in vivo. DN T cells profoundly inhibited the proliferation of CD4(+)CD25(-) T effector cells triggered by mDCs in vitro, and this suppression was further enhanced by IL-2. Adoptively transferring of DN T cells, in combination with IL-2, inhibited the proliferation and enhanced apoptosis of alloreactive CD4(+) T cells, which resulted in significant prolongation of skin allograft survival time. Perforin plays a key role in the enhancement of DN T cells immune regulation by IL-2. In conclusion, we elucidated that IL-2 promoted DN T cell proliferation and suppressive function. The combination of DN T cells and exogenous IL-2 may represent a novel therapy in the clinical setting to prevent allograft rejection and induce immune tolerance. PMID:27135902

  15. Retinoic acid signaling in mammalian eye development

    OpenAIRE

    CVEKL, ALES; Wang, Wei-Lin

    2009-01-01

    Retinoic acid (RA) is a biologically active metabolite of vitamin A (retinol) that serves as a signaling molecule during a number of developmental and physiological processes. RA signaling plays multiple roles during embryonic eye development. RA signaling is initially required for reciprocal interactions between the optic vesicle and invaginating lens placode. RA signaling promotes normal development of the ventral retina and optic nerve through its activities in the neural crest cell-derive...

  16. NOD-scid IL2R gamma(null) mice engrafted with human peripheral blood mononuclear cells as a model to test therapeutics targeting human signaling pathways

    OpenAIRE

    Zadeh-Khorasani, Maryam; Nolte, Thomas; Mueller, Thomas D.; Pechlivanis, Markos; Rueff, Franziska; Wollenberg, Andreas; Fricker, Gert; Wolf, Eckhard; Siebeck, Matthias; Gropp, Roswitha

    2013-01-01

    Background: Animal models of human inflammatory diseases have limited predictive quality for human clinical trials for various reasons including species specific activation mechanisms and the immunological background of the animals which markedly differs from the genetically heterogeneous and often aged patient population. Objective: Development of an animal model allowing for testing therapeutics targeting pathways involved in the development of Atopic Dermatitis (AD) with better transla...

  17. Association of mast cell-derived VEGF and proteases in Dengue shock syndrome.

    Directory of Open Access Journals (Sweden)

    Takahisa Furuta

    Full Text Available BACKGROUND: Recent in-vitro studies have suggested that mast cells are involved in Dengue virus infection. To clarify the role of mast cells in the development of clinical Dengue fever, we compared the plasma levels of several mast cell-derived mediators (vascular endothelial cell growth factor [VEGF], soluble VEGF receptors [sVEGFRs], tryptase, and chymase and -related cytokines (IL-4, -9, and -17 between patients with differing severity of Dengue fever and healthy controls. METHODOLOGY/PRINCIPAL FINDINGS: The study was performed at Children's Hospital No. 2, Ho Chi Minh City, and Vinh Long Province Hospital, Vietnam from 2002 to 2005. Study patients included 103 with Dengue fever (DF, Dengue hemorrhagic fever (DHF, and Dengue shock syndrome (DSS, as diagnosed by the World Health Organization criteria. There were 189 healthy subjects, and 19 febrile illness patients of the same Kinh ethnicity. The levels of mast cell-derived mediators and -related cytokines in plasma were measured by ELISA. VEGF and sVEGFR-1 levels were significantly increased in DHF and DSS compared with those of DF and controls, whereas sVEGFR-2 levels were significantly decreased in DHF and DSS. Significant increases in tryptase and chymase levels, which were accompanied by high IL-9 and -17 concentrations, were detected in DHF and DSS patients. By day 4 of admission, VEGF, sVEGFRs, and proteases levels had returned to similar levels as DF and controls. In-vitro VEGF production by mast cells was examined in KU812 and HMC-1 cells, and was found to be highest when the cells were inoculated with Dengue virus and human Dengue virus-immune serum in the presence of IL-9. CONCLUSIONS: As mast cells are an important source of VEGF, tryptase, and chymase, our findings suggest that mast cell activation and mast cell-derived mediators participate in the development of DHF. The two proteases, particularly chymase, might serve as good predictive markers of Dengue disease severity.

  18. Enhanced Th1-biased immune efficacy of porcine circovirus type 2 Cap-protein-based subunit vaccine when coadministered with recombinant porcine IL-2 or GM-CSF in mice.

    Science.gov (United States)

    Wang, Yiping; Lu, Yuehua; Liu, Dan; Wei, Yanwu; Guo, Longjun; Wu, Hongli; Huang, Liping; Liu, Jianbo; Liu, Changming

    2015-02-01

    Porcine circovirus type 2 (PCV2) capsid (Cap) protein is the primary protective antigen responsible for inducing PCV2-specific protective immunity, so it is a desirable target for the development of recombinant subunit vaccines to prevent PCV2-associated diseases. Interleukin 2 (IL-2) and granulocyte-macrophage colony-stimulating factor (GM-CSF), used as immune adjuvants, have been shown to enhance the immunogenicity of certain antigens or vaccines in various experimental models. In this study, five different subunit vaccines (the PCV2-Cap, Cap-PoIL-2, PCV2-Cap + PoIL-2, Cap-PoGM-CSF, and PCV2-Cap + PoGM-CSF vaccines) were prepared based on baculovirus-expressed recombinant proteins. The immunogenicity of these vaccines was evaluated to identify the immunoenhancement by PoIL-2 and PoGM-CSF of the Cap-protein-based PCV2 subunit vaccine in mice. The PCV2-Cap + PoIL-2, Cap-PoGM-CSF, PCV2-Cap + PoGM-CSF, and PCV2-Cap vaccines induced significantly higher levels of PCV2-specific antibodies than the Cap-PoIL-2 vaccine, whereas there was no apparent difference between these four vaccines. Our results indicate that neither PoIL-2 nor PoGM-CSF had effect on the enhancement of the humoral immunity induced by the PCV2-Cap vaccine. Furthermore, the PCV2-Cap + PoIL-2, Cap-PoGM-CSF, and PCV2-Cap + PoGM-CSF vaccines elicited stronger lymphocyte proliferative responses and greater IL-2 and interferon gamma (IFN-γ) secretion. This suggests that PoIL-2 and PoGM-CSF substantially augmented the Th1-biased immune response to the PCV2-Cap vaccine. Following challenge, the viral loads in the lungs of the PCV2-Cap + PoIL-2-, Cap-PoGM-CSF-, and PCV2-Cap + PoGM-CSF-treated groups were dramatically lower than those in the Cap-PoIL-2- and PCV2-Cap-treated groups, indicating that the three vaccines induced stronger protective effects against challenge. These findings show that PoIL-2 and PoGM-CSF essentially enhanced the Th1-biased protective efficacy of the

  19. 早期梅毒患者血清IL-2、IL-10水平变化的研究%Serum Levels of Interleukin-2 and- 10 in Patients with Early Syphilis

    Institute of Scientific and Technical Information of China (English)

    张秀英; 陈懿德; 庞玉森; 张景生; 严静丽

    2003-01-01

    Objective: To investigate the roles of interleukin-2(IL-2) and interleukin-10 (IL-10) in pathogenesis of early syphilis.Methods: The serum levels of IL-2 and IL-10 in 48patients with early syphilis were detected by ABCELISA.Results: (1) The level of IL-2 in the patients with early syphilis was significantly higher than that in healthy controls, while that of IL-10 was lower (P<0.001 and P<0.001). (2) The levels of IL-2 and IL10 were almost identical in patients with primary and secondary syphilis (P>0.05), as well as between different RPR titers (P>0.05). (3) After therapy, the level of IL-2 decreased markedly (P<0.05), while that of IL10 increase (p>0.05). (4) A significant correlation was found between the serum levels of IL-2 and IL-10 (r=0.5385 P<0.05).Conclusions: Th1 up-regulation occurs in patients with early syphilis, and plays an active role in fighting against TP infection.%目的 探讨细胞因子IL-2和IL-10在早期梅毒发病中的作用.方法 采用ABC-ELISA技术检测48例早期梅毒患者血清IL-2和IL-10水平.结果 (1)早期梅毒患者血清IL-2水平明显高于正常对照组,IL-10水平明显低于正常对照组(P值均<0.001).(2)一期梅毒患者血清IL-2和IL-10水平与二期梅毒无明显差异(P值均>0.05),不同R P R滴度间IL-2和IL-10水平无明显差异(P值均>0.05).(3)治疗后IL-2水平明显下降(P<0.05),IL-10水平上升不明显(P> 0.05).(4)IL-2和IL10之间呈正相关(r=0.5 3 8 5,P<0.05).结论 早期梅毒出现Th1上调,在抵御TP感染过程中发挥积极作用.

  20. The study on pathogenesis, TNF-α and sIL-2R changes in acute Guillain-Barré syndrome%急性Guillain-Barré综合征外周血TNF-α及sIL-2R的变化及发病机制探讨

    Institute of Scientific and Technical Information of China (English)

    华耀松; 钟述猷

    2002-01-01

    目的探讨肿瘤坏死因子(tumor necrosis factor-α,TNF-α)及可溶性白介素2受体(solubleinterleukin-2 receptor,sIL-2R)在急性Guillain-Barré综合征(Guillain-Barré syndrome,GBS)免疫发病机制中的作用.方法采用双抗体夹心ELISA法,对30例急性GBS患者外周血TNF-α及sIL-2R水平进行检测,并与24例其他神经系统疾病患者及正常对照组进行比较.结果急性GBS患者外周血TNF-α及sIL-2R水平明显升高,与正常对照组及其他神经系统疾病组比较差异有显著性意义(P<0.01);且外周血TNF-α水平的明显增高与GBS的病情进展有关.结论急性GRS患者有多种细胞因子异常,TNF-α、sIL-2R可能参与了急性GBS的免疫发病机制过程.

  1. Experimental Study on HSPgp96 and rIL-2 in the treatment of H22 Hepatocarcinoma Transplanted in Mice%HSPgp96与rIL-2治疗小鼠H22肝癌移植瘤的实验研究

    Institute of Scientific and Technical Information of China (English)

    施海燕; 周欣阳; 张天一; 林琳; 吴坚; 顾君一

    2004-01-01

    目的:研究热休克蛋白gp96(Heat-Shock Protein gp96,HSPgp96)-肽复合物瘤苗与重组人白介素-2(Recombinant Human Interleukin-2 ,rIL-2)对小鼠H22肝癌移植瘤的治疗效果.方法:从H22肝癌细胞中提取和纯化HSPgp96.建立小鼠H22肝癌移植瘤模型,应用HSPgp96与rIL-2治疗并比较疗效,观察指标:瘤重,光镜、电镜下观察组织结构.结果:经鉴定获得纯化的HSPgp96 .HSPgp96 5μg与rIL-2 25ku治疗小鼠H22肝癌有效.结论:该实验方法提纯的HSPgp96与重组人IL-2都能有效抑制小鼠H22肝癌移植瘤的生长.

  2. Reduced Immunogenicity of Induced Pluripotent Stem Cells Derived from Sertoli Cells

    OpenAIRE

    Xiaoying Wang; Jie Qin; Robert Chunhua Zhao; Martin Zenke

    2014-01-01

    Sertoli cells constitute the structural framework in testis and provide an immune-privileged environment for germ cells. Induced pluripotent stem cells (iPS cells) resemble embryonic stem cells (ES cells) and are generated from somatic cells by expression of specific reprogramming transcription factors. Here, we used C57BL/6 (B6) Sertoli cells to generate iPS cells (Ser-iPS cells) and compared the immunogenicity of Ser-iPS cells with iPS cells derived from mouse embryonic fibroblast (MEF-iPS ...

  3. Serum IL-2, IL-6, IL-10 and TNF-α Levels in Patients with Vitiligo before and after Treatment%白癜风患者治疗前后血清IL-2、IL-6、IL-10和TNF-α水平

    Institute of Scientific and Technical Information of China (English)

    孙岩; 钱立

    2011-01-01

    目的:探讨白癜风患者治疗前后血清IL-2、IL-6、IL-10、和TNF-α水平及临床意义.方法:应用放射免疫分析法检测32例白癜风患者治疗前后血清IL-2、IL-6、IL-10和TNF-α水平,以35名健康体检者为对照.结果:白癜风患者在治疗前血清IL-6、IL-10、TNF-α水平非常显著地高于对照组(P<0.01),IL-2水平非常显著地低于对照组(P<0.01);经治疗3个月后与对照组比较仍有显著性差异(P<0.05),且血清IL-2水平与IL-6、IL-10、TNF-α水平呈负相关(r=-0.482 2、-0.501 8、-0.613 4.P<0.01).结论:白癜风患者存在自身免疫调节的异常,检测血清IL-2、IL-6、IL-10和TNF-α水平的变化对疾病的治疗和预后具有重要的临床价值.%Objective: To detect serum levels of IL-2, IL-6, IL-10, and TNF-a in patients with vitili-go and to investigate its clinical significance. Methods:Radioimmunoassay was adopted to detect serum levels of IL-2, IL-6, IL-10 and TNF-α of 32 patients with vitiligo before and after treatment. The results were compared with those of 35 healthy people. Results: Serum levels of IL-6, IL-10, TNF-a in vitiligo patients before treatment were significantly higher than those of healthy people ( P<0.01) , while IL-2 level was significantly lower than that of healthy people (P<0.01). After treatment for 3 months, the differences between the two groups were significant (P<0. 05 ) , and serum IL-2 level was negatively correlated with IL-6, IL-10 and TNF-a levels (r=-0.482 2, -0.5018, -0.613 4.P <0.01). Conclusion: Autoimmune regulation is abnormal in vitiligo patients. Detection of serum IL-2, IL-6, IL-10, IL-6 and TNF-a levels have important clinical value in treatment and prognosis of the disease.

  4. Chemotherapy and anti-angiogenic drugs affect composition and coagulant phenotype of cell-derived vesicles in cancer patients

    NARCIS (Netherlands)

    Kleinjan, A.; Verhoeff, J.; Berckmans, R.; Kunst, P.; Van Doormaal, F.; Di Nisio, M.; Richel, D.; Kamphuisen, P.W.; Büller, H.R.; Nieuwland, R.

    2013-01-01

    Background: The relationship between chemotherapy and circulating microparticles in patients with cancer is complex. First, release of cancer cell-derived microparticles may contribute to resistance of cancer cells to chemotherapy. Second, chemotherapy and angiogenesis inhibiting agents promote a pr

  5. Effects of lamivudine on the function of dendritic cells derived from patients with chronic hepatitis B virus infection

    OpenAIRE

    Zheng, Peng-Yuan; Zhang, Dong-Yun; Lu, Gao-Feng; Yang, Ping-Chang; Qi, Yuan-Ming; Wang, Bai-Sheng

    2007-01-01

    AIM: To investigate if the nucleoside analogue lamivudine (LAM), a potent inhibitor of HBV replication, could restore the function of dendritic cells derived from patients with chronic hepatitis B (CHB) in an Asian population.

  6. Clinical significance of the Changes of Serum Levels of IL-2,IL-6,IL-10,IL-18 and T cell subset in patients with chronic nephritis%慢性肾炎治疗前后血清IL-2、IL-6、IL-10、IL-18和T淋巴细胞亚群检测意义

    Institute of Scientific and Technical Information of China (English)

    刘薇娜; 鲍培玉; 蒋全

    2010-01-01

    目的 探讨慢性肾炎患者治疗前后血清IL-2、IL-6、IL-10、IL-18和T淋巴细胞亚群的变化.方法 分别应用放免法、ELISA法和单克隆抗体法对30例慢性肾炎患者治疗前后进行了血清IL-2、IL-6、IL-10、IL-18和T淋巴细胞亚群水平的检测,并与35名正常健康人作比较.结果 慢性肾炎患者在治疗前血清IL-2和CD4/CD8比值明显低于正常人组(P<0.05),而IL-6、IL-10和IL-18水平高于正常人组(P<0.01);经半年治疗后血清IL-2、IL-6、IL-10、IL-18和CD4/CD8与治疗前组比较差异有统计学意义(P<0.05).结论 检测慢性肾炎患者血清IL-2、IL-6、IL-10、IL-18和T淋巴细胞亚群水平对判断病情及其预后均具有一定的临床实用价值.

  7. Activity-dependent long-term plasticity of afferent synapses on grafted stem/progenitor cell-derived neurons.

    OpenAIRE

    Toft Sörensen, Andreas; Rogelius, Nina; Lundberg, Cecilia; Kokaia, Merab

    2011-01-01

    Stem cell-based cell replacement therapies aiming at restoring injured or diseased brain function ultimately rely on the capability of transplanted cells to promote functional recovery. The mechanisms by which stem cell-based therapies for neurological conditions can lead to functional recovery are uncertain, but structural and functional repair appears to depend on integration of transplanted cell-derived neurons into neuronal circuitries. The nature by which stem/progenitor cell-derived neu...

  8. IL-2基因佐剂强化梅毒螺旋体外膜蛋白Tp92 DNA疫苗的免疫保护性研究%Evaluation of the protective effect of Treponema pallidum Tp92 DNA vaccine induced by genetic adjuvant IL-2

    Institute of Scientific and Technical Information of China (English)

    赵飞骏; 张晓红; 吴华; 曾铁兵; 刘双全; 曹训宇; 陈曦; 余坚

    2013-01-01

    目的 观察IL-2基因佐剂与梅毒螺旋体(Treponema pallidum,Tp)膜蛋白Tp92抗原编码基因DNA疫苗联合免疫新西兰兔后对Tp皮肤感染的免疫保护效果. 方法 将体重约3kg的新西兰兔随机分为pcDNA3.1(+)/IL-2+ pcDNA3.1(+)/Tp92联合免疫组,pcDNA3.1(+)/Tp92免疫组,pcDNA3.1(+)空质粒组及PBS空白组共4组,每组18只,每隔2周免疫一次,共免疫3次,初次免疫后第10周进行Tp皮肤感染,观察并记录感染早期各组家兔皮损情况;ELISA检测感染后不同时间点免疫兔血清特异性抗体水平和脾细胞IL-2及IFN-γ水平;MTT法检测感染后不同时间点兔脾淋巴细胞增殖水平. 结果 IL-2+ Tp92 DNA疫苗联合免疫组在Tp感染后0~224 d期间的不同时间点均检测到高滴度的特异性IgG,与Tp92 DNA疫苗组及各对照组比较差异均有统计学意义(P<0.05);感染后第14 d联合免疫组兔脾细胞培养上清中IFN-γ和IL-2水平均显著高于Tp92单独免疫组(P<0.05)及空质粒对照组和空白对照组(P<0.01),感染后不同时间点兔脾细胞均有明显增殖反应,SI值显著高于同期Tp92单独免疫组(P<0.05)及空质粒对照组和空白对照组(P<0.01).Tp感染早期联合免疫组家兔皮损Tp阳性率为20%,溃疡病灶形成率为10%,皮损愈合时间为48 d. 结论 IL-2基因佐剂和Tp92 DNA联合免疫能有效地诱导新西兰兔在Tp感染期间产生稳定持久保护性免疫应答,并能阻止Tp皮肤感染部位病损的发展.%Objective To estimate the protection efficacy of Tp92 DNA vaccine-induced immune responses enhanced by an interleukin (IL)-2 expression plasmid as a genetic adjuvant in a rabbit Treponema pallidum (Tp) skin challenge model. Methods New Zealand White rabbits were divided into four groups (pIL-2 + pTp92, pTp92, pcD, and PBS groups) that were immunized with three intramuscular inoculations of Tp92 DNA vaccine alone or in combination with a plasmid expressing IL-2 at 2-week intervals. Ten

  9. Detection of Levels of IL-2/IL-10 and Promoting Th2 Molecular IL-4 in Peripheral Blood of Gestational Hypertension%妊娠高血压外周血中促Th2的细胞因子水平及IL-2/IL-10平衡的临床意义

    Institute of Scientific and Technical Information of China (English)

    肖文辉; 钟荣钟; 林洁; 彭耀金

    2011-01-01

    目的:检测妊娠高血压患者外周血中促Th2的分子IL-4、IL-2与IL-10的水平,探讨IL-2/IL-10在妊高症中的临床意义.方法:选择40例未妊娠妇女为对照组,30例正常妊娠妇女为妊娠组,28例妊娠高血压患者为妊娠高血压组,ELISA检测血清中IL-4、IL-2和IL-10的水平.结果:与对照组外周血中IL-4水平(0.53±0.04)pg/ml相比:正常妊娠组IL-4水平升高至(0.91±0.03)pg/ml(P<0.05),妊娠高血压组II-4水平(0.67±0.35)pg/ml升高但明显低于正常妊娠组(P<0.01).与对照组外周血中IL-2水平(0.41±0.05)pg/ml相比:正常妊娠组IL-2水平升高至(0.82±0.11)pg/ml(P<0.01);妊娠高血压组IL-2水平高达1.57±0.22(pg/ml)明显高于其它两组(P<0.01).妊娠高血压组外周血中IL-10水平明显低于正常妊娠组IL-10水平(P<0.01);妊娠高血压组外周血中IL-2/IL-10比值明显高于于对照组及正常妊娠组的比值.结论:妊娠高血压患者外周血中细胞因子IL-2和IL-10分泌异常且诱导Th2细胞产生的IL-4降低,打破Th1/Th2平衡,致使Th1型免疫反应增强,使早孕期滋养细胞受到免疫损伤以致侵入能力下降,导致妊娠期高血压疾病的发生.%Objective: To detect the levels of IL-4 and IL-2 and IL-10 in the patients with pregnancy-induced hypertension, and discuss the detection of IL-2/IL-10. Methods: Taking 40 cases of non-pregnant women as control group, 30 normal pregnant women as pregnancy group, 28 cases of pregnancy induced hypertension as pregnancy-induced hypertension group patients, we used ELISA to detect the levels of IL-4, IL-2 and IL-10 in their serum. Results: The levels of IL-4 in peripheral blood of pregnancy group was (0.91± 0.03) pg / ml, significantly higher than the level of IL-4 (0.53 ± 0.04) pg / ml in control group (P<0.05). The levels of IL-4 in peripheral blood of pregnancy-induced hypertension group was (0.67± 0.35) pg / ml, significantly lower than the IL-4 level in normal pregnancy group (P

  10. Glial cell derived neurotrophic factor induces spermatogonial stem cell marker genes in chicken mesenchymal stem cells.

    Science.gov (United States)

    Boozarpour, Sohrab; Matin, Maryam M; Momeni-Moghaddam, Madjid; Dehghani, Hesam; Mahdavi-Shahri, Naser; Sisakhtnezhad, Sajjad; Heirani-Tabasi, Asieh; Irfan-Maqsood, Muhammad; Bahrami, Ahmad Reza

    2016-06-01

    Mesenchymal stem cells (MSCs) are known with the potential of multi-lineage differentiation. Advances in differentiation technology have also resulted in the conversion of MSCs to other kinds of stem cells. MSCs are considered as a suitable source of cells for biotechnology purposes because they are abundant, easily accessible and well characterized cells. Nowadays small molecules are introduced as novel and efficient factors to differentiate stem cells. In this work, we examined the potential of glial cell derived neurotrophic factor (GDNF) for differentiating chicken MSCs toward spermatogonial stem cells. MSCs were isolated and characterized from chicken and cultured under treatment with all-trans retinoic acid (RA) or glial cell derived neurotrophic factor. Expression analysis of specific genes after 7days of RA treatment, as examined by RT-PCR, proved positive for some germ cell markers such as CVH, STRA8, PLZF and some genes involved in spermatogonial stem cell maintenance like BCL6b and c-KIT. On the other hand, GDNF could additionally induce expression of POU5F1, and NANOG as well as other genes which were induced after RA treatment. These data illustrated that GDNF is relatively more effective in diverting chicken MSCs towards Spermatogonial stem cell -like cells in chickens and suggests GDNF as a new agent to obtain transgenic poultry, nevertheless, exploitability of these cells should be verified by more experiments.

  11. A pure population of lung alveolar epithelial type II cells derived from human embryonic stem cells.

    Science.gov (United States)

    Wang, Dachun; Haviland, David L; Burns, Alan R; Zsigmond, Eva; Wetsel, Rick A

    2007-03-13

    Alveolar epithelial type II (ATII) cells are small, cuboidal cells that constitute approximately 60% of the pulmonary alveolar epithelium. These cells are crucial for repair of the injured alveolus by differentiating into alveolar epithelial type I cells. ATII cells derived from human ES (hES) cells are a promising source of cells that could be used therapeutically to treat distal lung diseases. We have developed a reliable transfection and culture procedure, which facilitates, via genetic selection, the differentiation of hES cells into an essentially pure (>99%) population of ATII cells (hES-ATII). Purity, as well as biological features and morphological characteristics of normal ATII cells, was demonstrated for the hES-ATII cells, including lamellar body formation, expression of surfactant proteins A, B, and C, alpha-1-antitrypsin, and the cystic fibrosis transmembrane conductance receptor, as well as the synthesis and secretion of complement proteins C3 and C5. Collectively, these data document the successful generation of a pure population of ATII cells derived from hES cells, providing a practical source of ATII cells to explore in disease models their potential in the regeneration and repair of the injured alveolus and in the therapeutic treatment of genetic diseases affecting the lung. PMID:17360544

  12. Induced pluripotent stem cell-derived neural stem cell therapies for spinal cord injury

    Institute of Scientific and Technical Information of China (English)

    Corinne A Lee-Kubli; Paul Lu

    2015-01-01

    The greatest challenge to successful treatment of spinal cord injury is the limited regenerative capacity of the central nervous system and its inability to replace lost neurons and severed axons following injury. Neural stem cell grafts derived from fetal central nervous system tissue or embryonic stem cells have shown therapeutic promise by differentiation into neurons and glia that have the potential to form functional neuronal relays across injured spinal cord segments. However, implementation of fetal-derived or embryonic stem cell-derived neural stem cell ther-apies for patients with spinal cord injury raises ethical concerns. Induced pluripotent stem cells can be generated from adult somatic cells and differentiated into neural stem cells suitable for therapeutic use, thereby providing an ethical source of implantable cells that can be made in an autologous fashion to avoid problems of immune rejection. This review discusses the therapeutic potential of human induced pluripotent stem cell-derived neural stem cell transplantation for treatment of spinal cord injury, as well as addressing potential mechanisms, future perspectives and challenges.

  13. Functions of Müller cell-derived vascular endothelial growthfactor in diabetic retinopathy

    Institute of Scientific and Technical Information of China (English)

    2015-01-01

    Müller cells are macroglia and play many essentialroles as supporting cells in the retina. To respond topathological changes in diabetic retinopathy (DR), amajor complication in the eye of diabetic patients,retinal Müller glia produce a high level of vascularendothelial growth factor (VEGF or VEGF-A). As VEGFis expressed by multiple retinal cell-types and Müllerglia comprise only a small portion of cells in the retina,it has been a great challenge to reveal the function ofVEGF or other globally expressed proteins produced byMüller cells. With the development of conditional genetargeting tools, it is now possible to dissect the functionof Müller cell-derived VEGF in vivo . By using conditionalgene targeting approach, we demonstrate that Müllerglia are a major source of retinal VEGF in diabetic miceand Müller cell-derived VEGF plays a significant role inthe alteration of protein expression and peroxynitration,which leads to retinal inflammation, neovascularization,vascular leakage, and vascular lesion, key pathologicalchanges in DR. Therefore, Müller glia are a potentialcellular target for the treatment of DR, a leading causeof blindness.

  14. Gene expression profiling in multipotent DFAT cells derived from mature adipocytes

    International Nuclear Information System (INIS)

    Highlights: → Adipocyte dedifferentiation is evident in a significant decrease in typical genes. → Cell proliferation is strongly related to adipocyte dedifferentiation. → Dedifferentiated adipocytes express several lineage-specific genes. → Comparative analyses using publicly available datasets boost the interpretation. -- Abstract: Cellular dedifferentiation signifies the withdrawal of cells from a specific differentiated state to a stem cell-like undifferentiated state. However, the mechanism of dedifferentiation remains obscure. Here we performed comparative transcriptome analyses during dedifferentiation in mature adipocytes (MAs) to identify the transcriptional signatures of multipotent dedifferentiated fat (DFAT) cells derived from MAs. Using microarray systems, we explored similarly expressed as well as significantly differentially expressed genes in MAs during dedifferentiation. This analysis revealed significant changes in gene expression during this process, including a significant reduction in expression of genes for lipid metabolism concomitantly with a significant increase in expression of genes for cell movement, cell migration, tissue developmental processes, cell growth, cell proliferation, cell morphogenesis, altered cell shape, and cell differentiation. Our observations indicate that the transcriptional signatures of DFAT cells derived from MAs are summarized in terms of a significant decrease in functional phenotype-related genes and a parallel increase in cell proliferation, altered cell morphology, and regulation of the differentiation of related genes. A better understanding of the mechanisms involved in dedifferentiation may enable scientists to control and possibly alter the plasticity of the differentiated state, which may lead to benefits not only in stem cell research but also in regenerative medicine.

  15. Atomic force microscopy combined with human pluripotent stem cell derived cardiomyocytes for biomechanical sensing.

    Science.gov (United States)

    Pesl, Martin; Pribyl, Jan; Acimovic, Ivana; Vilotic, Aleksandra; Jelinkova, Sarka; Salykin, Anton; Lacampagne, Alain; Dvorak, Petr; Meli, Albano C; Skladal, Petr; Rotrekl, Vladimir

    2016-11-15

    Cardiomyocyte contraction and relaxation are important parameters of cardiac function altered in many heart pathologies. Biosensing of these parameters represents an important tool in drug development and disease modeling. Human embryonic stem cells and especially patient specific induced pluripotent stem cell-derived cardiomyocytes are well established as cardiac disease model.. Here, a live stem cell derived embryoid body (EB) based cardiac cell syncytium served as a biorecognition element coupled to the microcantilever probe from atomic force microscope thus providing reliable micromechanical cellular biosensor suitable for whole-day testing. The biosensor was optimized regarding the type of cantilever, temperature and exchange of media; in combination with standardized protocol, it allowed testing of compounds and conditions affecting the biomechanical properties of EB. The studied effectors included calcium , drugs modulating the catecholaminergic fight-or-flight stress response such as the beta-adrenergic blocker metoprolol and the beta-adrenergic agonist isoproterenol. Arrhythmogenic effects were studied using caffeine. Furthermore, with EBs originating from patient's stem cells, this biosensor can help to characterize heart diseases such as dystrophies. PMID:27266660

  16. Data on importance of hematopoietic cell derived Lipocalin 2 against gut inflammation.

    Science.gov (United States)

    Saha, Piu; Singh, Vishal; Xiao, Xia; Yeoh, Beng San; Vijay-Kumar, Matam

    2016-09-01

    The data herein is related to the research article entitled "Microbiota-inducible innate immune siderophore binding protein Lipocalin 2 is critical for intestinal homeostasis" (Singh et al., 2016) [1]. In the present article, we monitored dextran sodium sulfate (DSS)-induced colitis development upon Lipocalin 2 (Lcn2) neutralization, and examined the survival of Lcn2 deficient (Lcn2KO) mice and their WT littermates upon DSS challenge. To dissect the relative contribution of immune and non-immune cells-derived Lcn2 in mediating protection against gut inflammation, we generated respective bone marrow chimera and evaluated their susceptibility to IL-10 receptor neutralization-induced chronic colitis. Neutralization of Lcn2 in WT mice resulted in exacerbated DSS-induced colitis. Notably, mice lacking Lcn2 exhibited 100% mortality whereas only 20% mortality was observed in WT mice upon DSS challenge. Further, data from bone marrow chimera showed that immune cell-derived Lcn2 is the major contributor in conferring protection against colitis. PMID:27500193

  17. Gene expression profiling in multipotent DFAT cells derived from mature adipocytes

    Energy Technology Data Exchange (ETDEWEB)

    Ono, Hiromasa [Laboratory of Cell and Tissue Biology, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-8510 (Japan); Database Center for Life Science (DBCLS), Research Organization of Information and Systems (ROIS), Faculty of Engineering Bldg.12 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan); Oki, Yoshinao [Laboratory of Cell and Tissue Biology, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-8510 (Japan); Bono, Hidemasa [Database Center for Life Science (DBCLS), Research Organization of Information and Systems (ROIS), Faculty of Engineering Bldg.12 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan); Kano, Koichiro, E-mail: kkano@brs.nihon-u.ac.jp [Laboratory of Cell and Tissue Biology, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-8510 (Japan)

    2011-04-15

    Highlights: {yields} Adipocyte dedifferentiation is evident in a significant decrease in typical genes. {yields} Cell proliferation is strongly related to adipocyte dedifferentiation. {yields} Dedifferentiated adipocytes express several lineage-specific genes. {yields} Comparative analyses using publicly available datasets boost the interpretation. -- Abstract: Cellular dedifferentiation signifies the withdrawal of cells from a specific differentiated state to a stem cell-like undifferentiated state. However, the mechanism of dedifferentiation remains obscure. Here we performed comparative transcriptome analyses during dedifferentiation in mature adipocytes (MAs) to identify the transcriptional signatures of multipotent dedifferentiated fat (DFAT) cells derived from MAs. Using microarray systems, we explored similarly expressed as well as significantly differentially expressed genes in MAs during dedifferentiation. This analysis revealed significant changes in gene expression during this process, including a significant reduction in expression of genes for lipid metabolism concomitantly with a significant increase in expression of genes for cell movement, cell migration, tissue developmental processes, cell growth, cell proliferation, cell morphogenesis, altered cell shape, and cell differentiation. Our observations indicate that the transcriptional signatures of DFAT cells derived from MAs are summarized in terms of a significant decrease in functional phenotype-related genes and a parallel increase in cell proliferation, altered cell morphology, and regulation of the differentiation of related genes. A better understanding of the mechanisms involved in dedifferentiation may enable scientists to control and possibly alter the plasticity of the differentiated state, which may lead to benefits not only in stem cell research but also in regenerative medicine.

  18. 膠原病肺の病態に関する研究 第2編 膠原病肺における血清中および気管支肺胞洗浄液中 soluble IL-2 receptor の検討

    OpenAIRE

    横田, 聡

    1992-01-01

    Abnormality of cellular immunity has been reported in the pathogenesis of collagen vascular diseases (CVD). IL-2 and IL-2 receptors play important roles as immunological regulation in CVD. In this study, the soluble forms of IL-2 receptor (sIL-2R) and IL-2 receptor positive lymphocytes (CD25) were analyzed to evaluate the role of IL-2R in the pathogenesis of interstitial pneumonia (IP) of CVD such as RA, SLE, polymyositis/dermatomyositis (PM/DM), and PSS and idiopathic interstitial pneumonia ...

  19. Xenogeneic graft-versus-host-disease in NOD-scid IL-2Rγnull mice display a T-effector memory phenotype.

    Directory of Open Access Journals (Sweden)

    Niwa Ali

    Full Text Available The occurrence of Graft-versus-Host Disease (GvHD is a prevalent and potentially lethal complication that develops following hematopoietic stem cell transplantation. Humanized mouse models of xenogeneic-GvHD based upon immunodeficient strains injected with human peripheral blood mononuclear cells (PBMC; "Hu-PBMC mice" are important tools to study human immune function in vivo. The recent introduction of targeted deletions at the interleukin-2 common gamma chain (IL-2Rγ(null, notably the NOD-scid IL-2Rγ(null (NSG and BALB/c-Rag2(null IL-2Rγ(null (BRG mice, has led to improved human cell engraftment. Despite their widespread use, a comprehensive characterisation of engraftment and GvHD development in the Hu-PBMC NSG and BRG models has never been performed in parallel. We compared engrafted human lymphocyte populations in the peripheral blood, spleens, lymph nodes and bone marrow of these mice. Kinetics of engraftment differed between the two strains, in particular a significantly faster expansion of the human CD45(+ compartment and higher engraftment levels of CD3(+ T-cells were observed in NSG mice, which may explain the faster rate of GvHD development in this model. The pathogenesis of human GvHD involves anti-host effector cell reactivity and cutaneous tissue infiltration. Despite this, the presence of T-cell subsets and tissue homing markers has only recently been characterised in the peripheral blood of patients and has never been properly defined in Hu-PBMC models of GvHD. Engrafted human cells in NSG mice shows a prevalence of tissue homing cells with a T-effector memory (T(EM phenotype and high levels of cutaneous lymphocyte antigen (CLA expression. Characterization of Hu-PBMC mice provides a strong preclinical platform for the application of novel immunotherapies targeting T(EM-cell driven GvHD.

  20. CLINICAL SIGNIFICANCE OF SERUM CYTOKINES IL-1β,sIL-2R,IL-6,TNF-α,AND IFN-v IN ACUTE CORONARY SYNDROME

    Institute of Scientific and Technical Information of China (English)

    Yan-ni Wang; Shao-min Che; Ai-qun Ma

    2004-01-01

    Objectives To explore serum cytokines levels (including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v) and their significance in patients with acute coronary syndrome (ACS) and the subsequent follow-ups, with attempt to estimate the role of various serum inflammatory markers in the diagnosis and assessment of ACS.Methods The study population include 40 patients with acute myocardial infarction (AMI), 40 patients with unstable angina pectoris (UAP), and 40 controls. Among the 80 patients, 60 patients attended a follow up 4 months later. Serum inflammatory markers including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v were measured by enzyme linked immunosorbent assay.Results Serum IL- 1 β, sIL-2R, IL-6, TNF-α were significantly higher in AMI group or UAP group compared to the control group and became significantly lower 4 months later in the follow-up patients. Serum levels of IFN-v shows no significant difference between AMI group or UAP group and controls, also showing no significant change when measured in follow up patients. There was no correlation between serum creatine kinase-MB isoenzyme levels and serum inflammatory markers either in UAP or AMI group. Furthermore, when divided into two subgroups using Wagner's QRS scoring system in the AMI group, there is no difference of each serum inflammatory marker between ≤ 6 scores group and > 6 scores group.Conclusion Serum levels of certain inflammatory markers may have some diagnostic value for ACS, and can be a useful marker reflecting disease stability.

  1. Fusion Expression and Immunity Evaluation of Chicken IL-2 with Antigen Epitopes from F Gene of Newcastle Disease Virus%鸡IL-2/NDV-F蛋白抗原表位融合基因的表达及免疫性初探

    Institute of Scientific and Technical Information of China (English)

    许发芝; 吴胜国; 余为一

    2011-01-01

    To explore the immune adjuvant effects of chicken IL-2 ( ChIL-2), a recombinant plasmid with ChlL-2 and multi-antigen epitopes from F gene of Newcastle disease virus (NDV-F) was built and expressed in prokaryotic cells to prevent and control the chicken ND disease. The recombinant chimeric gene of ChIL-2-linker-NDV-F constructed by chicken ChIL-2 gene linked multi-antigen epitopes gene of NDV F protein via a serine-rich linker by overlap-PCR method was cloned into prokaryotic expression vector PET-32a. After identification by sequencing, the recombinant ChIL-2-inker-NDV-F protein was expressed in E. coli BL21 through IPTG and purified with the Ni2+ affinity column. The expressed ChIL-2-linker-NDV-F protein was analyzed by SDS-PAGE, Western-blot and indirect ELISA, respectively. The chimeric gene of ChIL-2-linker-NDV-F was successfully constructed and cloned into PET-32a vector respectively. The expressed ChIL-2-linker-NDV-F protein was shown by a major band with a expected molecular weight about 48 kD on SDS-PAGE and Western-blot and accounted for almost 45 % of the total bacteria proteins, Serological assay by indirect ELISA showed that it reacted strongly and specifically with chicken serum of NDV infection. These results indicated that the chimeric gene encoding ChIL-2-linker-NDV-F could effectively express in prokaryotic cells and the expressed protein had high specificity and good immunogenicity.%研究鸡白细胞介素-2(chicken interleukin-2,ChlL-2)的免疫佐剂功能,构建ChlL-2与新城疫病毒F蛋白多抗原表位(NDV-F)的嵌合基因,以对鸡新城疫疫病进行防治.采用重叠延伸PCR方法通过基因柔性接头将ChlL-2基因和NDV-F多抗原表位基因构建成ChL-2-linker-NDV-F嵌合基因并克隆入PET-32a载体,经测序鉴定后,转化BL21大肠杆菌,IPTG诱导表达6×His融合蛋白,Ni2+亲和柱纯化,表达产物经SDS-PAGE、Western blot 和间接ELISA检测和鉴定.结果表明,实验成功构建并克隆了ChlL-2-linker

  2. Dynamics of IL-2 and IL-7 levels during Highly active antiretroviral therapy and their significance%中国HIV感染者IL-2和IL-7参与高效抗反转录病毒治疗的抗病毒免疫反应及其在病毒控制中的作用

    Institute of Scientific and Technical Information of China (English)

    Diallo MA; 陈霞; 郑煜煌; 何艳; 贺波; 周华英; 谌资; 罗艳; 曾飔

    2012-01-01

    Objective The objective of this study was to investigate the course of certain common gamma cytokines ( IL-2 and IL-7 ) and their role on the control of the viral infection in a short term antiviral therapy.Methods A total of 35 adults with chronic HIV infection,responding to combined antiretroviral therapy (cART) guideline criteria were enrolled in this one year follow-up study.After signing an informed consent,20 ml blood were collected from each patient at base line,week 0,week 24 and week 48.1 ml serum collected from each patient was kept at -80 * C until use.Serum concentration of IL-2 and IL-7 was determined using ELISA kit from "ebioscience Beijing".CD4 and CD8 cells were counted and quantified using flux cytometry,and serum HIV RNA was quantified using real time PCR.Results All patients had a mean baseline IL-2 level [ (9.67 ± 2.6 ) pg/ml ]lower than the controls [ ( 27.36 ± 5.05 ) pg/ml ].After treatment for 48 weeks,IL-2 increased[ ( 19.8 ± 3.3 ) pg/ml ].However,the mean baseline 1L-7 [ ( 81.74± 20.47 ) pg/ml ]in patients was higher than controls [ ( 2.06 ± 1.52 ) pg/ml ].After treatment for 48 weeks,IL-7 decreased [ (8.36 ± 2.16)pg/ml ].IL-2 showed a significant increase and positive correlation with CD4 cells after HAART initiation (0week:R =0.21,P =0.063,24week:R =0.24,P =0.033,48week:R =0.19,P =0.103; IL-7 showed a significant decrease after HAART initiation but it did not show correlation with CD4 cells.We noted there was a negative correlation between IL-2 and CD4 count in HAART baseline (R =0.28,P =0.012 ),but no correlation between IL-7 and CD4 count from 6 month after HAART.IL-2 showed negative correlation with HIV RNA ( R =- 0.17,P =0.032),but IL-7 showed a relationship with the HIV RNA Conclusions The increase of IL-2 coupled with the decrease of IL-7 revealed a partial restoration of immune response during HAART.However,the absence of relationship with HIV RNA suggested that these cytokines might not be directly involved in the reduction

  3. 中药"凤香洗液"对伴HPV感染的CIN患者宫颈局部IL-2和IL-4的影响%Effects of traditional Chinese medicine"Fengxiang Lotion"on local cervical IL-2 and IL-4 in patients of cervical intraepithelial neoplasia with HPV infection

    Institute of Scientific and Technical Information of China (English)

    李小宁; 贺丰杰; 吉喆

    2015-01-01

    Objective To observe the effects of traditional Chinese medical compound"Fengxiang Lotion"on the local vaginal immune state for patients of cervical intraepithelial neoplasia (CIN) with human papillomavirus (HPV) infection.Methods Sixty patients with high risk HPV infection diagnosed with cervical biopsy using colpos-copy in the Affiliated Hospital of Shaanxi University of Chinese Medicine and Xi'an XD Group Hospital from July 2013 to February 2015 were selected in the study,which were divided into CINⅠand CINⅡgroup (n=30 each).Thirty healthy women coming for medical examination during the same period served as the control group.CINⅠgroup received Fengxiang Lotion for treatment,while the other two groups not.The levels of interleukin (IL)-2 and IL-4 in cervical-vaginal lavage solution,which were synthesized by type 1 and type 2 helper T cells respective-ly,were tested by ELISA.Results (1) IL-2 levels of CINⅠgroup and CINⅡgroup were significantly lower than that of the control group (P<0.01),while IL-4 levels of CINⅠgroup and CINⅡ group were significantly higher (P<0.05).The ratios of IL-2 to IL-4 in CINⅠgroup and CINⅡgroup were significantly lower than that of the con-trol group (P<0.05).What's more,all these changes was more significant in CINⅡgroup than CINⅠgroup (P<0.05).(2) In CINⅠgroup,the IL-2 level increased significantly after medication (P<0.05),while IL-4 decreased significantly (P<0.05),with the ratio of IL-2 to IL-4 increased significantly (P<0.01).Conclusion The IL-2 lev-els and IL-2/IL-4 ratio in CIN patients with HPV decreased,while IL-4 increased.These changes were consistent with the severity of CIN."Fengxiang Lotion"may block the progression of CIN through regulating the balance of Th1 and Th2.%目的 观察中药"凤香洗液"对伴有HPV感染的CINⅠ、CINⅡ患者阴道局部免疫状态的影响及治疗效果.方法 选取2013年7月至2015年2月于陕西中医学院附属医院、西安西电集团医院门诊就

  4. Plasmodium berghei ANKA infection increases Foxp3, IL-10 and IL-2 in CXCL-10 deficient C57BL/6 mice

    Directory of Open Access Journals (Sweden)

    Stiles Jonathan K

    2011-03-01

    Full Text Available Abstract Background Cerebral malaria (CM is a major cause of malaria mortality. Sequestration of infected red blood cells and leukocytes in brain vessels coupled with the production of pro-inflammatory factors contribute to CM. CXCL-10 a chemokine that is chemotactic to T cells has been linked to fatal CM. Mice deficient for CXCL-10 gene are resistant to murine CM, while antibody ablation of CXCL-10 enhanced the production of regulatory T cells (CD4+Cd25+Foxp3+ and IL-10 which regulate the immune system. Interleukin-2 (IL-2, a pro-inflammatory cytokine implicated in malaria pathogenesis has also been shown to be a key regulator of Foxp3. However the role of Foxp3 in resistant murine CM is not well understood. Methods The hypothesis that resistance of CXCL-10-/- mice to murine CM may be due to enhanced expression of Foxp3 in concert with IL-10 and IL-2 was tested. CXCL-10-/- and WT C57BL/6 mice were infected with Plasmodium berghei ANKA and evaluated for CM symptoms. Brain, peripheral blood mononuclear cells (PBMCs and plasma were harvested from infected and uninfected mice at days 2, 4 and 8. Regulatory T cells (CD4+CD25+ and non-T regs (CD4+CD25- were isolated from PBMCs and cultured with P. berghei antigens in vitro with dendritic cells as antigen presenting cells. Regulatory T cell transcription and specific factor Foxp3, was evaluated in mouse brain and PBMCs by realtime-PCR and Western blots while IL-10, and IL-2 were evaluated in plasma and cultured supernatants by ELISA. Results Wild type mice exhibited severe murine CM symptoms compared with CXCL-10-/- mice. Foxp3 mRNA and protein in brain and PBMC's of CXCL-10-/- mice was significantly up-regulated (p P. berghei antigens produced more IL-10 than WT CD4+CD25+ T cells. Conclusion The results indicate that in the absence of CXCL-10, the resulting up-regulation of Foxp3, IL-10 and IL-2 may be involved in attenuating fatal murine CM.

  5. Acute exhaustive exercise regulates IL-2, IL-4 and MyoD in skeletal muscle but not adipose tissue in rats

    OpenAIRE

    Seelaender Marília; Santos Ronaldo VT; Pimentel Gustavo D; Oyama Lila M; Zanchi Nelo E; Lira Fábio S; Rosa Neto José C; Oller do Nascimento Cláudia M

    2011-01-01

    Abstract Background The purpose of this study was to evaluate the effect of exhaustive exercise on proteins associated with muscle damage and regeneration, including IL-2, IL-4 and MyoD, in extensor digitorum longus (EDL) and soleus muscles and mesenteric (MEAT) and retroperitoneal adipose tissues (RPAT). Methods Rats were killed by decapitation immediately (E0 group, n = 6), 2 (E2 group, n = 6) or 6 (E6 group, n = 6) hours after the exhaustion protocol, which consisted of running on a treadm...

  6. El factor de transferencia como inductor de la expresión de RNAm de IFN-γ e IL-2 en pollos vacunados contra influenza aviar

    Directory of Open Access Journals (Sweden)

    A Bravo-Blas

    2010-01-01

    Full Text Available La influenza aviar es una enfermedad de gran importancia económica para la industria avícola. En México sólo se ha reportado la cepa H5N2 de baja patogenicidad y ésta se controla mediante la vacunación con virus inactivado. Esta vacuna en emulsión reduce la presencia de signos, pero no la eliminación viral. Desde hace más de 50 años se ha informado acerca de la eficacia del Factor de Transferencia (FT como inmunomodulador en casos clínicos humanos y en menor cantidad en modelos animales. El objetivo de este trabajo fue el de establecer la dosis que produce un mayor porcentaje de expresión del RNAm de dos citocinas: IL-2 y de IFN-γ. Se diseñó un experimento para evidenciar la expresión del RNAm de estas dos citocinas en pollos previamente inoculados con FT específico para influenza aviar. En la primera fase se aplicaron 0,1, 1, y 10 unidades de FT a diferentes grupos de pollos, posteriormente se realizó la PCR a partir de tejido esplénico. En la segunda fase se aplicó el FT junto con la vacuna a tres nuevos grupos de pollos. Del experimento 1 solamente IL-2 tuvo un porcentaje mayor de positivos (58,33% con 1 unidad (P < 0,05. En cambio, en el experimento 2, con 1 unidad se obtuvo 75% de positivos para IL-2 (P < 0,05 y 100% para IFN-γ (P < 0,01. De estos resultados su puede concluir que al aplicar una unidad de FT (equivalente a 7,3 μg de proteína al inicio del experimento y 10 días después otra unidad de FT junto con la vacuna inactivada de IA se indujo la expresión del RNAm de IFN-γ e IL-2.

  7. Effect of Drug Sera from per os Da Cheng Qi(大承气) Granule on Production/Secretion of IL-2 by Intestinal Intraepithelial Lymphocytes of Mice%大承气颗粒药物血清对小鼠肠上皮内淋巴细胞产生/分泌IL-2的作用

    Institute of Scientific and Technical Information of China (English)

    方步武; 吴咸中; 来丽娜; 陈菲; 刘俊红; 李继坤; 王民宪; 崔志清; 林秀珍

    2005-01-01

    目的:探讨大承气颗粒药物血清对小鼠肠上皮内淋巴细胞产生/分泌白细胞介素-2(interleukin-2,IL-2)的作用.方法:以不同浓度的大承气颗粒剂灌胃大鼠后不同时间点于无菌条件下采取腹腔静脉血,制备药物血清;分离培养肠上皮内淋巴细胞(intestinal intraepithelial lymphocytes,IELs),以药物血清作用于IELs,设培养液对照、IEL对照、正常血清对照,采用放射免疫分析法测定IL-2.结果:不同浓度大承气颗粒的药物血清在多数时间点的作用明显强于培养液对照、IEL对照及正常大鼠血清对照.正常大鼠血清对照组与IEL对照组的IL-2水平很接近.同时间点的药物血清比较,在30 min和12 h时,4%大承气颗粒的药物血清作用强;在3 h时,40%大承气颗粒的作用最强.4%、20%大承气颗粒的药物血清使小鼠分泌IL-2的作用在1.5 h达峰值,在24 h出现第二个峰值;40%大承气颗粒药物血清使IEL分泌IL-2的峰值在3 h.结论:不同浓度的大承气颗粒制备的药物血清具有使IEL产生/分泌IL-2的作用;该作用涉及到药物本身和药物在体内的活性代谢物和/或其诱导的内源性活性物质的作用.

  8. Could a B-1 cell derived phagocyte "be one" of the peritoneal macrophages during LPS-driven inflammation?

    Directory of Open Access Journals (Sweden)

    Ana Flavia Popi

    Full Text Available The inflammatory response is driven by signals that recruit and elicit immune cells to areas of tissue damage or infection. The concept of a mononuclear phagocyte system postulates that monocytes circulating in the bloodstream are recruited to inflamed tissues where they give rise to macrophages. A recent publication demonstrated that the large increase in the macrophages observed during infection was the result of the multiplication of these cells rather than the recruitment of blood monocytes. We demonstrated previously that B-1 cells undergo differentiation to acquire a mononuclear phagocyte phenotype in vitro (B-1CDP, and we propose that B-1 cells could be an alternative origin for peritoneal macrophages. A number of recent studies that describe the phagocytic and microbicidal activity of B-1 cells in vitro and in vivo support this hypothesis. Based on these findings, we further investigated the differentiation of B-1 cells into phagocytes in vivo in response to LPS-induced inflammation. Therefore, we investigated the role of B-1 cells in the composition of the peritoneal macrophage population after LPS stimulation using osteopetrotic mice, BALB/Xid mice and the depletion of monocytes/macrophages by clodronate treatment. We show that peritoneal macrophages appear in op/op((-/- mice after LPS stimulation and exhibit the same Ig gene rearrangement (VH11 that is often found in B-1 cells. These results strongly suggest that op/op((-/- peritoneal "macrophages" are B-1CDP. Similarly, the LPS-induced increase in the macrophage population was observed even following monocyte/macrophage depletion by clodronate. After monocyte/macrophage depletion by clodronate, LPS-elicited macrophages were observed in BALB/Xid mice only following the transfer of B-1 cells. Based on these data, we confirmed that B-1 cell differentiation into phagocytes also occurs in vivo. In conclusion, the results strongly suggest that B-1 cell derived phagocytes are a component of

  9. Cervical cancer cell-derived interleukin-6 impairs CCR7-dependent migration of MMP-9-expressing dendritic cells.

    Science.gov (United States)

    Pahne-Zeppenfeld, Jennifer; Schröer, Nadine; Walch-Rückheim, Barbara; Oldak, Monika; Gorter, Arko; Hegde, Subramanya; Smola, Sigrun

    2014-05-01

    Cervical carcinogenesis is a consequence of persistent infection with high-risk human papillomaviruses (HPVs). Recent studies indicate that HPV-transformed cells actively instruct their microenvironment to promote carcinogenesis. Here, we demonstrate that cervical cancer cells activate monocytes to produce their own CCL2 for further monocyte recruitment and reprogram their function during differentiation and maturation to dendritic cells (DCs). Our data show that cervical cancer cells suppress the induction of the chemokine receptor CCR7 in phenotypically mature DCs and impair their migration toward a lymph node homing chemokine, required to initiate adaptive immune responses. We confirmed the presence of CD83(+)CCR7(low) DCs in cancer biopsies. The second factor essential for DC migration, matrix-metalloproteinase MMP-9, which also has vasculogenic and protumorigenic properties, is not suppressed but upregulated in immature as well as mature DCs. We identified interleukin-6 (IL-6) as a crucial cervical cancer cell-derived mediator and nuclear factor kappaB (NF-jB) as the central signaling pathway targeted in DCs. Anti-IL-6 antibodies reverted not only NF-jB inhibition and restored CCR7-dependent migration but also blocked MMP-9 induction. This is the first report demonstrating the dissociation of CCR7 and MMP-9 expression in phenotypically mature CD83(+) DCs by cancer cells. Our results show that cervical cancer cells actively shape the local microenvironment. They induce the accumulation of myeloid cells and skew their function from immune activation to local production of protumorigenic MMP-9. Neutralizing anti-IL-6 antibodies can counteract this functional dysbalance and should therefore be considered for adjuvant cervical cancer therapy.

  10. Effect of stromal-cell-derived factor 1 on stem-cell homing and tissue regeneration in ischaemic cardiomyopathy

    Science.gov (United States)

    Askari, Arman T.; Unzek, Samuel; Popovic, Zoran B.; Goldman, Corey K.; Forudi, Farhad; Kiedrowski, Matthew; Rovner, Aleksandr; Ellis, Stephen G.; Thomas, James D.; DiCorleto, Paul E.; Topol, Eric J.; Penn, Marc S.

    2003-01-01

    BACKGROUND: Myocardial regeneration via stem-cell mobilisation at the time of myocardial infarction is known to occur, although the mechanism for stem-cell homing to infarcted tissue subsequently and whether this approach can be used for treatment of ischaemic cardiomyopathy are unknown. We investigated these issues in a Lewis rat model (ligation of the left anterior descending artery) of ischaemic cardiomyopathy. METHODS: We studied the effects of stem-cell mobilisation by use of granulocyte colony-stimulating factor (filgrastim) with or without transplantation of syngeneic cells. Shortening fraction and myocardial strain by tissue doppler imaging were quantified by echocardiography. FINDINGS: Stem-cell mobilisation with filgrastim alone did not lead to engraftment of bone-marrow-derived cells. Stromal-cell-derived factor 1 (SDF-1), required for stem-cell homing to bone marrow, was upregulated immediately after myocardial infarction and downregulated within 7 days. 8 weeks after myocardial infarction, transplantation into the peri-infarct zone of syngeneic cardiac fibroblasts stably transfected to express SDF-1 induced homing of CD117-positive stem cells to injured myocardium after filgrastim administration (control vs SDF-1-expressing cardiac fibroblasts mean 7.2 [SD 3.4] vs 33.2 [6.0] cells/mm2, n=4 per group, pstem-cell homing to injured myocardium and suggest a strategy for directed stem-cell engraftment into injured tissues. Our findings also indicate that therapeutic strategies focused on stem-cell mobilisation for regeneration of myocardial tissue must be initiated within days of myocardial infarction unless signalling for stem-cell homing is re-established.

  11. Effect of a four-week exercise program on the secretion of IFN-γ, TNF-α, IL-2 and IL-6 cytokines in elite Taekwondo athletes

    Science.gov (United States)

    Kaya, Oktay

    2016-01-01

    The aim of the present study was to examine how a 4-week exercise program affects the serum levels of certain cytokines in Taekwondo athletes. The study involved 10 elite male Taekwondo athletes (mean age, 20.67±0.24 years; mean weight, 65.45±1.69 kg) who were studying at the Physical Education and Sports High School of Selçuk University (Konya, Turkey) in June 2014. The subjects were involved in a Taekwondo exercise program on every weekday for 4 weeks. The subjects were also engaged in an exercise to exhaustion session twice; once before starting the 4-week exercise program and once upon completion of the program. Blood samples were collected from the subjects in four rounds: During rest, upon fatigue, and before and after the 4-week exercise program. These samples were analyzed to establish the serum levels of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin (IL)-2 and IL-6 using enzyme-linked immunosorbent assay test kits. Pre- and post-exercise program, the IFN-γ and TNF-α levels did not show any significant difference. When compared with the pre-exercise levels, serum IL-2 levels of the subjects were found to be elevated after the 4-week exercise program. The highest serum IL-6 values were established after the subjects were exercised to fatigue before the exercise program was initiated (Pperformance.

  12. CD25 shedding by human natural occurring CD4+CD25+ regulatory T cells does not inhibit the action of IL-2

    DEFF Research Database (Denmark)

    Pedersen, Anders Elm; Lauritsen, Jens Peter Holst

    2009-01-01

    Regulatory T (Treg) cells are important for the maintenance of peripheral tolerance and inhibition of pathogenic T-cell responses. Therefore, they are important for the limitation of chronic inflammation but can also be deleterious by e.g. limiting antitumour immune responses. Natural occurring...... Tregs are known to inhibit CD4+ T cell in a contact-dependent manner, but at the same time, various suppressive factors are secreted. We, here, demonstrate that human naturally occurring CD4+CD25+ Tregs are able to shed large amounts of soluble CD25 upon activation. Secretion of sCD25 could add to the...... inhibitory effect of Tregs as such secretion in other settings has been proposed to act as a sink for local IL-2. However, we here demonstrate that supernatant from human Tregs containing high concentration of sCD25 does not inhibit proliferation of CD4+CD25(-) T cells or inhibit the action of IL-2 in an in...

  13. A New IL-2RG Gene Mutation in an X-linked SCID Identified through TREC/KREC Screening: a Case Report

    Directory of Open Access Journals (Sweden)

    Maryam Nourizadeh

    2015-10-01

    Full Text Available Severe combined immunodeficiency (SCID represents a rare group of primary immunodeficiency disorders (PIDs, with known or unknown genetic alterations. Here, we report a new interleukin 2 receptor, gamma chain (IL-2RG mutation in an Iranian SCID newborn.The patient was a 6-day old boy with a family history of PID. The child was screened using a molecular-based analysis for the assessment of T cell receptor excision circles (TRECs and kappa-deleting recombination excision circles (KRECs. Moreover, a complete immunological evaluation and gene sequencing was performed.Results showed undetectable TREC but a high level of KREC copy numbers. Flowcytometric data indicated low numbers of T and NK cells, but elevated number of B cells. A novel substitution in IL2RG: c.675 C>A, leading to p.225 Ser>Arg was found. Based on the functional analysis, the mutation is predicted to be damaging. The patient was diagnosed as a T B+ NK X-linked SCID.

  14. Efficient genetic manipulation of the NOD-Rag1-/-IL2RgammaC-null mouse by combining in vitro fertilization and CRISPR/Cas9 technology.

    Science.gov (United States)

    Li, Feng; Cowley, Dale O; Banner, Debra; Holle, Eric; Zhang, Liguo; Su, Lishan

    2014-06-17

    Humanized mouse models have become increasingly important and widely used in modeling human diseases in biomedical research. Immunodeficient mice such as NOD-Rag1-/-IL2RgammaC-null (NRG) or NOD-SCID-IL2RgammaC-null (NSG) mice are critical for efficient engraftment of human cells or tissues. However, their genetic modification remains challenging due to a lack of embryonic stem cells and difficulty in the collection of timed embryos after superovulation. Here, we report the generation of gene knockout NRG mice by combining in vitro fertilization (IVF) and CRISPR/Cas9 technology. Sufficient numbers of fertilized embryos were produced through IVF, and a high rate of Fah gene targeting was achieved with microinjection of Cas9 mRNA, gRNA and single strand oligonucleotide DNA (ssDNA) into the embryos. The technology paves the way to construct NRG or NSG mutant mice to facilitate new humanized mouse models. The technology can also be readily adapted to introduce mutations in other species such as swine and non-human primates.

  15. Thermal ablation versus conventional regional hyperthermia has greater anti-tumor activity against melanoma in mice by upregulating CD4~+ cells and enhancing IL-2 secretion

    Institute of Scientific and Technical Information of China (English)

    Yingying Zhang; Wei Zhang; Cuanying Geng; Tongjun Lin; Xiaowen Wang; Lingyun Zhao; Jintian Tang

    2009-01-01

    To determine whether conventional hyperthermia (42-45℃) or ablation therapy (>50℃) achieves better synergistic effects on direct cytotoxicity and anti-tumor immunity,we compared the therapeutic effects of two hyperthermia temperatures,43 and 55℃,in terms of cytotoxicity and upregulation of immune functions in a mouse malignant melanoma model.Melanoma-bearing mice were treated by directly applying regional hyperthermia to the tumor nodule with a heating light at a temperature of 43℃ for 30 min or 55℃ for 10 min.The tumor growth curve and mice survival rate were observed.To investigate the hyperthermia-induced immunological response,peripheral blood CD4~+ and CD8~+ T cells and the serum IL-2 level were determined.Our results indicated that application of regional hyperthermia at the ablation temperature (such as 55℃) achieved better synergistic anti-tumor effects than did conventional hyperthermia (43℃).Significant increases in the number of peripheral blood CD4~+ T cells and the serum IL-2 level likely contributed to the underlying mechanism.

  16. Blood Serum Levels of IL-2, IL-6, IL-8, TNF-α and IL-1β in Patients on Maintenance Hemodialysis

    Institute of Scientific and Technical Information of China (English)

    Jacek Rysz; Maciej Banach; Aleksandra Cialkowska-Rysz; Robert Stolarek; Marcin Barylski; Jaroslaw Drozdz; Piotr Okonski

    2006-01-01

    Cytokines are essential mediators of immune response and inflammatory reactions. Patients with chronic renal failure (CRF) commonly present with abnormalities of immune function related with impaired kidney function and the accumulation of uremic toxins in addition to bioincompatibility of dialyzer membranes. During a hemodialysis (HD) session, cytokines are released mainly by monocytes activated by endotoxin-type compounds in dialyzer fluid,complement factors and direct contact with dialyzer membrane. The study included 15 CRF patients, aged 36.4 ± 2.9 years, on regular HD maintenance therapy for mean 68 ± 10 months and 15 healthy controls. It was designed to assess serum levels of a panel of inflammatory cytokines: IL-1β, IL-2, IL-6, IL-8 and TNF-α in CRF patients on regular maintenance HD before, 20, 60 and 240 minutes of a single HD session in parallel with C-reactive protein (CRP) as an additional parameter. CRP concentration was increased in HD patients when compared with healthy controls. The concentrations of IL-1, IL-6, IL-8 and TNF-α were increased, whereas the serum level of IL-2 was not altered during a single HD session.

  17. Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Afford New Opportunities in Inherited Cardiovascular Disease Modeling

    Directory of Open Access Journals (Sweden)

    Daniel R. Bayzigitov

    2016-01-01

    Full Text Available Fundamental studies of molecular and cellular mechanisms of cardiovascular disease pathogenesis are required to create more effective and safer methods of their therapy. The studies can be carried out only when model systems that fully recapitulate pathological phenotype seen in patients are used. Application of laboratory animals for cardiovascular disease modeling is limited because of physiological differences with humans. Since discovery of induced pluripotency generating induced pluripotent stem cells has become a breakthrough technology in human disease modeling. In this review, we discuss a progress that has been made in modeling inherited arrhythmias and cardiomyopathies, studying molecular mechanisms of the diseases, and searching for and testing drug compounds using patient-specific induced pluripotent stem cell-derived cardiomyocytes.

  18. Effect of purine alkaloids on the proliferation of lettuce cells derived from protoplasts.

    Science.gov (United States)

    Sasamoto, Hamako; Fujii, Yoshiharu; Ashihara, Hiroshi

    2015-05-01

    To investigate the ecological role of caffeine, theobromine, theophylline and paraxanthine, which are released from purine alkaloid forming plants, the effects of these purine alkaloids on the division and colony formation of lettuce cells were assessed at concentrations up to 1 mM. Five days after treatment with 500 μM caffeine, theophylline and paraxanthine, division of isolated protoplasts was significantly inhibited. Thirteen days treatment with > 250 μM caffeine had a marked inhibitory effect on the colony formation of cells derived from the protoplasts. Other purine alkaloids also acted as inhibitors. The order of the inhibition was caffeine > theophylline > paraxanthine > theobromine. These observations suggest that a relatively low concentration of caffeine is toxic for proliferation of plant cells. In contrast, theobromine is a weak inhibitor of proliferation. Possible allelopathic roles of purine alkaloids in natural ecosystems are discussed.

  19. Induced pluripotent stem cell-derived cardiomyocytes: boutique science or valuable arrhythmia model?

    Science.gov (United States)

    Knollmann, Björn C

    2013-03-15

    This article reviews the strengths and limitations of induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) as models of cardiac arrhythmias. Specifically, the article attempts to answer the following questions: Which clinical arrhythmias can be modeled by iPSC-CM? How well can iPSC-CM model adult ventricular myocytes? What are the strengths and limitations of published iPSC-CM arrhythmia models? What new mechanistic insight has been gained? What is the evidence that would support using iPSC-CM to personalize antiarrhythmic drug therapy? The review also discusses the pros and cons of using the iPSC-CM technology for modeling specific genetic arrhythmia disorders, such as long QT syndrome, Brugada Syndrome, or Catecholaminergic Polymorphic Ventricular Tachycardia. PMID:23569106

  20. Induced pluripotent stem cell-derived cardiomyocytes: boutique science or valuable arrhythmia model?

    Science.gov (United States)

    Knollmann, Björn C

    2013-03-15

    This article reviews the strengths and limitations of induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) as models of cardiac arrhythmias. Specifically, the article attempts to answer the following questions: Which clinical arrhythmias can be modeled by iPSC-CM? How well can iPSC-CM model adult ventricular myocytes? What are the strengths and limitations of published iPSC-CM arrhythmia models? What new mechanistic insight has been gained? What is the evidence that would support using iPSC-CM to personalize antiarrhythmic drug therapy? The review also discusses the pros and cons of using the iPSC-CM technology for modeling specific genetic arrhythmia disorders, such as long QT syndrome, Brugada Syndrome, or Catecholaminergic Polymorphic Ventricular Tachycardia.

  1. Genetic strategies to investigate neuronal circuit properties using stem cell-derived neurons

    Directory of Open Access Journals (Sweden)

    Isabella eGarcia

    2012-12-01

    Full Text Available The mammalian brain is anatomically and functionally complex, and prone to diverse forms of injury and neuropathology. Scientists have long strived to develop cell replacement therapies to repair damaged and diseased nervous tissue. However, this goal has remained unrealized for various reasons, including nascent knowledge of neuronal development, the inability to track and manipulate transplanted cells within complex neuronal networks, and host graft rejection. Recent advances in embryonic stem cell (ESC and induced pluripotent stem cell (iPSC technology, alongside novel genetic strategies to mark and manipulate stem cell-derived neurons now provide unprecedented opportunities to investigate complex neuronal circuits in both healthy and diseased brains. Here, we review current technologies aimed at generating and manipulating neurons derived from ESCs and iPSCs towards investigation and manipulation of complex neuronal circuits, ultimately leading to the design and development of novel cell-based therapeutic approaches.

  2. Failure of low-dose recombinant human IL-2 to support the survival of virus-specific CTL clones infused into severe combined immunodeficient foals: lack of correlation between in vitro activity and in vivo efficacy.

    Science.gov (United States)

    Mealey, Robert H; Littke, Matt H; Leib, Steven R; Davis, William C; McGuire, Travis C

    2008-01-15

    Although CTL are important for control of lentiviruses, including equine infectious anemia virus (EIAV), it is not known if CTL can limit lentiviral replication in the absence of CD4 help and neutralizing antibody. Adoptive transfer of EIAV-specific CTL clones into severe combined immunodeficient (SCID) foals could resolve this issue, but it is not known whether exogenous IL-2 administration is sufficient to support the engraftment and proliferation of CTL clones infused into immunodeficient horses. To address this question we adoptively transferred EIAV Rev-specific CTL clones into four EIAV-challenged SCID foals, concurrent with low-dose aldesleukin (180,000U/m2), a modified recombinant human IL-2 (rhuIL-2) product. The dose was calculated based on the specific activity on equine PBMC in vitro, and resulted in plasma concentrations considered sufficient to saturate high affinity IL-2 receptors in humans. Despite specific activity on equine PBMC that was equivalent to recombinant equine IL-2 and another form of rhuIL-2, aldesleukin did not support the engraftment and expansion of infused CTL clones, and control of viral load and clinical disease did not occur. It was concluded that survival of Rev-specific CTL clones infused into EIAV-challenged SCID foals was not enhanced by aldesleukin at the doses used in this study, and that in vitro specific activity did not correlate with in vivo efficacy. Successful adoptive immunotherapy with CTL clones in immunodeficient horses will likely require higher doses of rhuIL-2, co-infusion of CD4+ T lymphocytes, or administration of equine IL-2. PMID:17727961

  3. Rett syndrome induced pluripotent stem cell-derived neurons reveal novel neurophysiological alterations.

    Science.gov (United States)

    Farra, N; Zhang, W-B; Pasceri, P; Eubanks, J H; Salter, M W; Ellis, J

    2012-12-01

    Rett syndrome (RTT) is a neurodevelopmental autism spectrum disorder caused by mutations in the methyl-CpG-binding protein 2 (MECP2) gene. Here, we describe the first characterization and neuronal differentiation of induced pluripotent stem (iPS) cells derived from Mecp2-deficient mice. Fully reprogrammed wild-type (WT) and heterozygous female iPS cells express endogenous pluripotency markers, reactivate the X-chromosome and differentiate into the three germ layers. We directed iPS cells to produce glutamatergic neurons, which generated action potentials and formed functional excitatory synapses. iPS cell-derived neurons from heterozygous Mecp2(308) mice showed defects in the generation of evoked action potentials and glutamatergic synaptic transmission, as previously reported in brain slices. Further, we examined electrophysiology features not yet studied with the RTT iPS cell system and discovered that MeCP2-deficient neurons fired fewer action potentials, and displayed decreased action potential amplitude, diminished peak inward currents and higher input resistance relative to WT iPS-derived neurons. Deficiencies in action potential firing and inward currents suggest that disturbed Na(+) channel function may contribute to the dysfunctional RTT neuronal network. These phenotypes were additionally confirmed in neurons derived from independent WT and hemizygous mutant iPS cell lines, indicating that these reproducible deficits are attributable to MeCP2 deficiency. Taken together, these results demonstrate that neuronally differentiated MeCP2-deficient iPS cells recapitulate deficits observed previously in primary neurons, and these identified phenotypes further illustrate the requirement of MeCP2 in neuronal development and/or in the maintenance of normal function. By validating the use of iPS cells to delineate mechanisms underlying RTT pathogenesis, we identify deficiencies that can be targeted for in vitro translational screens.

  4. MicroRNA and protein profiling of brain metastasis competent cell-derived exosomes.

    Directory of Open Access Journals (Sweden)

    Laura Camacho

    Full Text Available Exosomes are small membrane vesicles released by most cell types including tumor cells. The intercellular exchange of proteins and genetic material via exosomes is a potentially effective approach for cell-to-cell communication and it may perform multiple functions aiding to tumor survival and metastasis. We investigated microRNA and protein profiles of brain metastatic (BM versus non-brain metastatic (non-BM cell-derived exosomes. We studied the cargo of exosomes isolated from brain-tropic 70W, MDA-MB-231BR, and circulating tumor cell brain metastasis-selected markers (CTC1BMSM variants, and compared them with parental non-BM MeWo, MDA-MB-231P and CTC1P cells, respectively. By performing microRNA PCR array we identified one up-regulated (miR-210 and two down-regulated miRNAs (miR-19a and miR-29c in BM versus non-BM exosomes. Second, we analyzed the proteomic content of cells and exosomes isolated from these six cell lines, and detected high expression of proteins implicated in cell communication, cell cycle, and in key cancer invasion and metastasis pathways. Third, we show that BM cell-derived exosomes can be internalized by non-BM cells and that they effectively transport their cargo into cells, resulting in increased cell adhesive and invasive potencies. These results provide a strong rationale for additional investigations of exosomal proteins and miRNAs towards more profound understandings of exosome roles in brain metastasis biogenesis, and for the discovery and application of non-invasive biomarkers for new therapies combating brain metastasis.

  5. Scaling and automation of a high-throughput single-cell-derived tumor sphere assay chip.

    Science.gov (United States)

    Cheng, Yu-Heng; Chen, Yu-Chih; Brien, Riley; Yoon, Euisik

    2016-10-01

    Recent research suggests that cancer stem-like cells (CSCs) are the key subpopulation for tumor relapse and metastasis. Due to cancer plasticity in surface antigen and enzymatic activity markers, functional tumorsphere assays are promising alternatives for CSC identification. To reliably quantify rare CSCs (1-5%), thousands of single-cell suspension cultures are required. While microfluidics is a powerful tool in handling single cells, previous works provide limited throughput and lack automatic data analysis capability required for high-throughput studies. In this study, we present the scaling and automation of high-throughput single-cell-derived tumor sphere assay chips, facilitating the tracking of up to ∼10 000 cells on a chip with ∼76.5% capture rate. The presented cell capture scheme guarantees sampling a representative population from the bulk cells. To analyze thousands of single-cells with a variety of fluorescent intensities, a highly adaptable analysis program was developed for cell/sphere counting and size measurement. Using a Pluronic® F108 (poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol)) coating on polydimethylsiloxane (PDMS), a suspension culture environment was created to test a controversial hypothesis: whether larger or smaller cells are more stem-like defined by the capability to form single-cell-derived spheres. Different cell lines showed different correlations between sphere formation rate and initial cell size, suggesting heterogeneity in pathway regulation among breast cancer cell lines. More interestingly, by monitoring hundreds of spheres, we identified heterogeneity in sphere growth dynamics, indicating the cellular heterogeneity even within CSCs. These preliminary results highlight the power of unprecedented high-throughput and automation in CSC studies.

  6. Chronic inhibition of tumor cell-derived VEGF enhances the malignant phenotype of colorectal cancer cells

    International Nuclear Information System (INIS)

    Vascular endothelial growth factor-a (VEGF)-targeted therapies have become an important treatment for a number of human malignancies. The VEGF inhibitors are actually effective in several types of cancers, however, the benefits are transiently, and the vast majority of patients who initially respond to the therapies will develop resistance. One of possible mechanisms for the acquired resistance may be the direct effect(s) of VEGF inhibitors on tumor cells expressing VEGF receptors (VEGFR). Thus, we investigated here the direct effect of chronic VEGF inhibition on phenotype changes in human colorectal cancer (CRC) cells. To chronically inhibit cancer cell-derived VEGF, human CRC cell lines (HCT116 and RKO) were chronically exposed (2 months) to an anti-VEGF monoclonal antibody (mAb) or were disrupted the Vegf gene (VEGF-KO). Effects of VEGF family members were blocked by treatment with a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI). Hypoxia-induced apoptosis under VEGF inhibited conditions was measured by TUNEL assay. Spheroid formation ability was assessed using a 3-D spheroid cell culture system. Chronic inhibition of secreted/extracellular VEGF by an anti-VEGF mAb redundantly increased VEGF family member (PlGF, VEGFR1 and VEGFR2), induced a resistance to hypoxia-induced apoptosis, and increased spheroid formation ability. This apoptotic resistance was partially abrogated by a VEGFR-TKI, which blocked the compensate pathway consisted of VEGF family members, or by knockdown of Vegf mRNA, which inhibited intracellular function(s) of all Vegf gene products. Interestingly, chronic and complete depletion of all Vegf gene products by Vegf gene knockout further augmented these phenotypes in the compensate pathway-independent manner. These accelerated phenotypes were significantly suppressed by knockdown of hypoxia-inducible factor-1α that was up-regulated in the VEGF-KO cell lines. Our findings suggest that chronic inhibition of tumor cell-derived VEGF

  7. Suppression of Th1-mediated autoimmunity by embryonic stem cell-derived dendritic cells.

    Directory of Open Access Journals (Sweden)

    Tokunori Ikeda

    Full Text Available We herein demonstrate the immune-regulatory effect of embryonic stem cell-derived dendritic cells (ES-DCs using two models of autoimmune disease, namely non-obese diabetic (NOD mice and experimental autoimmune encephalomyelitis (EAE. Treatment of pre-diabetic NOD mice with ES-DCs exerted almost complete suppression of diabetes development during the observation period for more than 40 weeks. The prevention of diabetes by ES-DCs was accompanied with significant reduction of insulitis and decreased number of Th1 and Th17 cells in the spleen. Development of EAE was also inhibited by the treatment with ES-DCs, and the therapeutic effect was obtained even if ES-DCs were administrated after the onset of clinical symptoms. Treatment of EAE-induced mice with ES-DCs reduced the infiltration of inflammatory cells into the spinal cord and suppressed the T cell response to the myelin antigen. Importantly, the ES-DC treatment did not affect T cell response to an exogenous antigen. As the mechanisms underlying the reduction of the number of infiltrating Th1 cells, we observed the inhibition of differentiation and proliferation of Th1 cells by ES-DCs. Furthermore, the expression of VLA-4α on Th1 cells was significantly inhibited by ES-DCs. Considering the recent advances in human induced pluripotent stem cell-related technologies, these results suggest a clinical application for pluripotent stem cell-derived dendritic cells as a therapy for T cell-mediated autoimmune diseases.

  8. Effects of tacrolimus on morphology, proliferation and differentiation of mesenchymal stem cells derived from gingiva tissue

    Science.gov (United States)

    HA, DONG-HO; YONG, CHUL SOON; KIM, JONG OH; JEONG, JEE-HEON; PARK, JUN-BEOM

    2016-01-01

    Tacrolimus is a 23-membered macrolide lactone with potent immunosuppressive activity that is effective in the prophylaxis of organ rejection following kidney, heart and liver transplantation. Tacrolimus also exerts a variety of actions on bone metabolism. The aim of the present study was to evaluate the effects of different concentrations of tacrolimus on the morphology and viability of human stem cells derived from the gingiva. Gingival-derived stem cells were grown in the presence of tacrolimus at final concentrations ranging from 0.001 to 100 µg/ml. The morphology of the cells was viewed under an inverted microscope and the cell viability was analyzed using Cell Counting kit-8 (CCK-8) on days 1, 3, 5 and 7. Alizarin Red S staining was used to assess mineralization of treated cells. The control group showed spindle-shaped, fibroblast-like morphology and the shapes of the cells in 0.001, 0.01, 0.1, 1 and 10 µg/ml tacrolimus were similar to those of the control group. All groups except the 100 µg/ml group showed increased cell proliferation over time. Cultures grown in the presence of tacrolimus at 0.001, 0.01, 0.1, 1 and 10 µg/ml were not identified to be significantly different compared with the control at days 1, 3 and 5 using the CCK-8 assays. Increased mineralized deposits were noted with increased incubation time. Treatment with tacrolimus from 0.001 to 1 µg/ml led to an increase in mineralization compared with the control group. Within the limits of this study, tacrolimus at the tested concentrations (ranging from 0.001 to 10 µg/ml) did not result in differences in the viability of stem cells derived from gingiva; however it did enhance osteogenic differentiation of the stem cells. PMID:27177273

  9. DETECTION OF PBMC T CELL SUBSETS AND mIL-2R OF THE PERSONS INFECTED BY CRYPTOSPORIDIUM PARVUM BY USING THE SYSTEM OF BSA%应用BSA系统检测隐孢子虫感染者外周血T细胞亚群及mIL-2R

    Institute of Scientific and Technical Information of China (English)

    许礼发; 李朝品; 张荣波; 王晓秋

    2004-01-01

    Objective To explore the changes of cellular immune function in the persons infected by Cryptosporidium parvum .MethodsThe system of biotin streptavidin (BSA) was used to detect T cell subsets and membrane interleukin 2 receptor (mIL 2R) in peripheral blood mononuclear cell (PBMC) of the persons infected byC. parvum . ResultsThe positive rates of CD3+,CD4+,CD8+ and CD4+/CD8+ in the persons in whose stoolsC. parvum oocysts were found were (54.77±7.33)%,(38.17±7.52)%,(30.64±5.87)% and 1.28±0.72 respectively,and the level of mIL 2R in induction period was (32.15±2.42)%. It could be concluded that the positive rates of CD3+,CD4+,CD8+ and CD4+/CD8+ in the persons in whose stoolsC. parvum oocysts were found were significantly different from those with noC. parvum oocysts were found (P <0.05~0.01). The level of mIL 2R in induction period was significantly different from which in resting period (P <0.05~0.01).ConclusionIn the persons infected byC. parvum,cellular immunity participates in resisting the parasite,and T cell subsets and mIL 2R play an important role.%目的探讨隐孢子虫感染者机体细胞免疫功能的变化. 方法采用生物素-链霉亲和素(Biotin-Streptavidin , BSA)法对卵囊阳性患者分别进行外周血T细胞亚群、膜白介素-2受体(mIL-2R)检测. 结果隐孢子虫卵囊阳性者的CD3+、CD4+、CD8+百分率和CD4+/CD8+比值分别为(54.77±7.33)%、(38.17±7.52)%、(30.64±5.87)% 和1.28±0.72,诱导期mIL-2R表达水平为(32.15±2.42)%,两者分别与卵囊阴性者及静息期mIL-2R相比,差异有显著性(P<0.05~0.01). 结论隐孢子虫感染者引起细胞免疫功能的变化,T细胞亚群、mIL-2R在抗隐孢子虫感染中起重要作用.

  10. 姜黄素对戊四氮致癫大鼠海马区 IL-2和IL-6表达水平的影响%Effect of the curcumin on expression of IL-2 and IL-6 of hippocampus in pentyle-netetrazol-induced epilepsy in rats

    Institute of Scientific and Technical Information of China (English)

    董伟; 严建维; 谈巧玲; 叶森

    2016-01-01

    Objective To investigate the mechanism of anti-epileptic effect of the curcumin .Methods The SD rats were injected intraperitoneally with pentylenetetrazol kindling 25 .0 mg/kg to induce a rat epilepsy model .All of the treatments were performed once a day continuously for 28 days .The rats in blank group and model group received 5 ml of normal saline .The rats in the high and low curcumin group were given 200 mg/kg and 100 mg/kg of curcumin once a day ,respectively .The rats in the sodium valproate (VPA) group were given 400 mg/kg of VPA once a day by gavage .After treatment ,the seizures level was recorded by using the Racine′s six point grading scale ,and the expression of IL-2 and IL-6 of hippocampus were detected by the enzyme linked immunoassay (ELISA) .Results The seizures level was reduced by curcumin in epileptic rats .The ex-pressions of IL-2 and IL-6 of the model group were significantly higher than those of the blank group (P0 .05) . Conclusion The curcumin can reduce the seizure level in rats ,it shows some anti-epileptic effets and dose-dependently ,which may be through down-regulating the expression of IL-2 and IL-6 in hippocampus .%目的:探讨姜黄素抗癫的作用机制。方法取健康成年雄性SD大鼠,连续腹腔注射戊四氮,诱发大鼠点燃致癫模型。空白组和模型组灌予生理盐水5 ml ,1次/d ,连续28 d。低剂量和高剂量姜黄素组分别灌予姜黄素100 mg/kg和200 mg/kg ,1次/d ,连续28 d;丙戊酸钠组灌予丙戊酸钠400 mg/kg ,1次/d ,连续28 d。治疗结束后,按照Racine的6级评分标准,观察癫大鼠发作等级变化,用酶联免疫法(ELISA)检测海马区IL-2和IL-6表达水平的变化。结果姜黄素组的癫大鼠惊厥发作等级降低。模型组IL-2和IL-6高表达,比空白组呈显著升高(P<0.05)。与模型组比较,姜黄素组大鼠海马区IL-2和IL-6的表达明显降低( P<0.05);与低剂量姜黄素组

  11. IL-2, IFN-γ, IL-4、 IL-6 Expression in Peri-implant Gingival Crevicular Fluid and Clinical Significance%种植体周围炎龈沟液中IL-2、 IFN-γ、IL-4、IL-6的表达及临床意义

    Institute of Scientific and Technical Information of China (English)

    郦兴

    2015-01-01

    目的 探讨探讨白细胞介素2、4、6(IL-2、IL-4、IL-6)、干扰素γ(IFN-γ)在种植体周围炎龈沟液(GCF)中的表达及意义.方法 选择2011年6月~2014年6月50例种植体周围炎患者为研究对象.另选择健康种植体患者50例和健康人群50例为对照组.统计3组探诊深度(PD)、龈沟出血指数(SBI)、IL-2、IFN-γ、IL-4、IL-6水平.结果 种植体周围炎组PD、SBI、GCF均高于健康种植体组和对照组,差异有统计学意义(P<0.01);但健康种植体组和对照组PD、SBI、GCF差异无统计学意义(P>0.05);种植体周围炎组IL-2、IFN-γ水平低于健康种植体组和对照组(P<0.01),健康种植体组IL-2、IFN-γ水平低于对照组(P<0.05),种植体周围炎组IL-4、IL-6水平高于健康种植体组和对照组(P<0.01),健康种植体组IL-4、IL-6水平高于对照组(P<0.05);≥60岁组IL-2、IFN-γ水平低于<60岁组(P<0.01),≥60岁组IL-4、IL-6水平高于<60岁组(P<0.01);种植体周围炎患者GCF中IL-2与IFN-γ呈正相关(r=4.267,P=0.016),与IL-4、IL-6分别呈负相关(r=4.352、4.615,P=0.005、0.002),IFN-γ与IL-4、IL-6分别呈负相关(r=4.322、4.603,P=0.009、0.005),IL-4,IL-6呈正相关(r=4.065,P=0.019).结论 种植体周围炎患者IL-4、IL-6高表达,IL-2、IFN-γ低表达,IL-2、IFN-γ、IL-4、IL-6的表达与种植体周围炎患者性别无关,与种植体周围炎患者的年龄有关.

  12. 过敏性哮喘患者外周血IL-2、IL-4、IL-6和IL-8水平的变化%Change of the Level of IL-2, IL-4, IL-6 and IL-8 of Irritability Asthma Patients

    Institute of Scientific and Technical Information of China (English)

    王爱华; 李全新

    2000-01-01

    为了观察白细胞介素在哮喘发病中的作用,该文分别对哮喘发作期20例 ,缓解期12例患者及正常对照组10例进行了血清IL-2、IL-4、IL-6和IL-8的测定,结果发现 ,发作期患者血清IL-2水平下降,而IL-4、IL-6和IL-8水平均增高,缓解期均恢复至正常水平.提示这些细胞因子在哮喘发病过程中起着重要作用.

  13. Treatment of refractory non-Hodgkin's lymphoma of the spleen with combined autologous cytokine induced killer cells and IL-2: A case report%自体CIK细胞联合IL-2治疗难治性脾非霍奇金淋巴瘤1例

    Institute of Scientific and Technical Information of China (English)

    杨洋; 陈云燕; 张文英; 刘洋; 王瑶; 代汉仁; 韩为东; 张峰; 姚善谦; 杨波; 脱帅; 卢学春; 朱宏丽; 脱朝伟; 蔡力力; 迟小华; 于睿莉

    2012-01-01

    目的 观察自体细胞因子诱导的杀伤细胞输注联合IL-2治疗老年淋巴瘤的有效性和安全性.方法 1例老年脾脏恶性淋巴瘤患者在经过手术切除脾脏原发肿瘤、8个疗程的R-CHOP方案免疫化疗后出现淋巴瘤肝脏转移,此时采集患者外周血单核细胞,在体外经干扰素-γ (IFN-γ)、白介素-2(IL-2)和抗CD3单克隆抗体诱导成CIK细胞,每次回输细胞数为2-3×109个,回输后应用IL-2 100mU/d,皮下注射,连续10d,28d为1个周期.观察治疗前后患者肝功能、肿瘤相关生物学指标及影像学变化.结果 患者共完成8个周期的CIK细胞输注,每次回输后未出现不良反应,肝功能指标、LDH水平均降至正常,PET/CT检查示治疗后肝脏淋巴瘤转移灶消失,达到完全缓解.结论 自体CIK细胞输注联合IL-2疗法对于清除治疗后淋巴瘤的微小残留灶和(或)转移灶安全有效.%Objective To observe the efficiency and safety of combined autologous cytokine induced killer(CIK) cells and IL-2 in treatment of splenic lymphoma in old patients. Methods Hepatic metastasis of splenic primary malignant lymphoma occurred in an old patient after splenectomy and 8 cycles of R-CHOP chemotherapy. Peripheral blood mononuclear cells (PBMC) were collected. CIK cells were induced with in vitro interferon gamma (IFN-γ), IL-2 and anti-CD3 monoclonal antibody (mAb). Liver function, tumor-related biological indexes and image changes were observed after 2-3×109 CIK cells were re-transfused into the patient each time and IL-2 100mU/d was subcutaneously injected for 10 days, 28 days a cycle. Results No adverse reaction occurred in the patient after 8 cycles of CIK cells transfusion. The liver function and serum LDH level became normal(P<0.05). PET-CT showed that hepatic metastasis of lymphoma disappeared and completely relieved. Conclusion Combined autologous CIK cells and IL-2 is safe and effective for small residual or metastatic foci of lymphoma in old

  14. 性病患者58例血清IL-2、IL-6和IL-8水平的检测及其临床意义

    Institute of Scientific and Technical Information of China (English)

    高英举

    2002-01-01

    目的探讨血清白细胞介素-2(IL-2)、白细胞介素-6(IL-6),白细胞介素-8(IL-8)在性病患者血清中水平及临床意义.方法应用双抗体夹心EliSA法检测了58例性病患者血清中IL-2、IL-6和IL-8水平.结果 58例性病患者血清中,IL-2、IL-6、IL-8水平分别为(6.4±3.2) ng/ml、(0.15±0.06) ng/ml、(0.34±0.13) ng/ml、而IL-8则显著高于正常人组(P<0.01),其中IL-2、IL-6则明显低于正常人组(P<0.01).结论性病患者的发生与发展与IL-2、IL-6降低和IL-8升高密切相关,检测IL-2、IL-6和IL-8血清水平有助于性病的判断和治疗的选择.

  15. Enhancement of anti-CD4 mAb on anti-tumor activity of tumor-specific T cells activated by rIL-2 and anti-CD3 mAb%抗 C D 4单抗增强抗 C D 3单抗 /IL-2活化的肿瘤特异性 T细胞的抗瘤活性

    Institute of Scientific and Technical Information of China (English)

    黄辉; 俞红; 熊思东; 林云璐; 秦慧莲

    2001-01-01

    Aim To explore enhancing effects of anti-CD4 mAb on theproliferation and anti-tumor activity of tumor-specific T cells stimulated by anti-CD3 mAb. Methods Four different procedures were applied to culture splenocytes from tumor cell-immunized mice:(1)cultured in 2× 104U/L rIL-2 alone(IL-2 group);(2) cultured in anti-CD3 mAb alone (anti-CD3 group); (3) activated by anti-CD3 mAb for 48 h and then cultured in anti-CD3 mAb plus 2× 104U/L rIL-2 (anti-CD3+ IL-2 group); (4) activated by anti-CD3 and anti-CD4 mAbs for 48 h and then cultured in anti-CD3 and anti-CD4 mAbs plus 2× 104U/L rIL-2(anti-CD3+ IL-2+ anti-CD4 group). Effects of anti-CD4 mAb on proliferation, anti-tumor activity and phenotype of tumor-specific T cells were studied by 3H-TdR incorporation, 3H-TdR realease assay and FACS respectvely. Results 3H-TdR incorporation(cpm) in the cultured splenocytes from anti-CD3+ IL-2+ anti-CD4 group were 46 193, 31 047 and 7 443 on day 6, 12 and 20 respectively, while 3H-TdR incorporation(cpm) in the cultured splenocytes from anti-CD3+ IL-2 group were 22 045, 13 986 and 1 931 on day 6,12 and 20 respectively. The maximum anti-tumor activities of both group cells were 83.6% and 91.7% respectively on day 12. The cell phenotype analysis by FACS on day 12 indicated that phenotype of more than 99% effective cells in anti-CD3+ IL-2+ anti-CD4 group were Thy1.2+ . The percentage of CD4+ and CD25+ T cells in anti-CD3+ IL-2+ anti-CD4 group was also higher than that in anti-CD3+ IL-2 group. Conclusion Anti-CD4 mAb may enhance the proliferation and anti-tumor activity of tumor-specific T cells activated by rIL-2 and anti-CD3 mAb.%目的探讨抗CD4mAb增强抗CD3mAb刺激的肿瘤特异性T细胞增殖和杀瘤活性的作用。方法将肿瘤细胞免疫的小鼠脾细胞,采用4种不同的方案培养:(1)单独加2×104U/LrIL-2(IL-2组);(2)单独加抗CD3mAb(抗CD3组);(3)加抗CD3mAb48h后,再加入抗CD3mAb和2×104U/LrIL-2(抗CD3+IL-2

  16. 人肝再生增强因子对体外单个核细胞PKCα-NF-κB及IL-2的影响%Effects of human augmenter of liver regeneration on PKCα-NF-κB expressions and IL-2 secretion in peripheral mononuclear cells

    Institute of Scientific and Technical Information of China (English)

    王新国; 刘杞; 孙航

    2009-01-01

    目的 观察人肝再生增强因子(human augmenter of liver regeneration,hALR)对人外周血单个核细胞(periph-eral blood mononuclear cell,PBMC)PKCα-NF-κB及IL-2的影响,探讨hALR免疫抑制机制.方法 用梯度离心法分离正常人PBMC,分成正常对照组、刀豆蛋白A(ConA)刺激组(ConA组)和hALR干预组(ConA+hALR组).ConA组用5μg/mlConA刺激细胞增殖和分泌细胞因子IL-2;ConA+hALR组用30μg/ml hALR对ConA刺激进行干预.于细胞培养0、8、16、32、40 h时,用MTT法测定细胞增殖程度,Weston blot检测细胞内信号通路PKCα-NF-κB表达含量,Fluo-3AM装载检测胞内钙离子浓度,用双抗夹心酶联法检测上清细胞因子IL-2含量.结果 ConA组细胞增殖逐渐加强,16 h时与正常对照组比较具有统计学意义(P<0.05);PKCα、NF-κB表达上调及上清IL-2含量增加趋势一致,在16 h达到高峰(P<0.05).ConA+hALR组细胞没有增殖,PKCα-NF-κB表达及上清IL-2含量最高峰后移至32 h.每个时间点各组细胞内Ca2+浓度没有差别.结论 hALR可能通过抑制PKCα-NF-κB通路阻碍IL-2的产生速度进而抑制免疫.%Objective To observe the effects of human Augmenter of liver regeneration (hALR) on the expressions of PKCα, NF-ΚB and IL-2 in human peripheral blood mononuclear cells ( PBMC) and to evaluate the immunosuppressive mechanism of hALR. Methods PBMCs were isolated from healthy volunteers by gradi-ent centrifugation and were divided into normal control group (N), ConA stimulated group ( ConA group) and hALR interfering group ( ConA + hALR group). Cells in the ConA group were stimulated by ConA (5 μg/ml) to proliferate and to secret IL-2. ConA + hALR group was interfered by hALR at the dose of 30 μg/ml. After cells were cultured for 0, 8, 16, 32 and 40 h, the cell proliferation was detected by MTT assay. The expre-ssions of PKCa and NF-ΚB were detected by Western blot analysis. The concentration of intracellular calcium was determined according to

  17. The expression of IL-2 and IFN-γ of premature ovarian failure in murine autoimmune oophoritis immunized by procine zona pellucida%实验性自身免疫性卵巢功能早衰中IL-2和IFN-γ的检测

    Institute of Scientific and Technical Information of China (English)

    杨桂华; 柯丹

    2007-01-01

    目的:探讨Th1型细胞因子IL-2和IFN-γ在自身免疫性卵巢功能早衰(POF)中的表达.方法:分别用猪卵透明带(PZP)抗原和PBS缓冲液注射免疫Balb/c小鼠,共免疫3次,每2W免疫1次,经过3次免疫后,观察各组卵巢组织病理形态,并测定各组IL-2和IFN-γ水平.结果:PZP免疫小鼠卵巢组织形态明显改变,且Th1性细胞因子IL-2和IFN-γ表达均明显高于对照组.结论:自身免疫性卵巢功能早衰与T细胞介导的Th1型免疫应答有关.

  18. Primordial germ cell migration in the yellowtail kingfish (Seriola lalandi) and identification of stromal cell-derived factor 1.

    Science.gov (United States)

    Fernández, J A; Bubner, E J; Takeuchi, Y; Yoshizaki, G; Wang, T; Cummins, S F; Elizur, A

    2015-03-01

    Primordial germ cells (PGCs) are progenitors of the germ cell lineage, giving rise to either spermatogonia or oogonia after the completion of gonadal differentiation. Currently, there is little information on the mechanism of PGCs migration leading to the formation of the primordial gonad in perciform fish. Yellowtail kingfish (Seriola lalandi) (YTK) (order Perciforms) inhabit tropical and temperate waters in the southern hemisphere. Fundamental details into the molecular basis of larval development in this species can be easily studied in Australia, as they are commercially cultured and readily available. In this study, histological analysis of YTK larvae revealed critical time points for the migration of PGCs to the genital ridge, resulting in the subsequent development of the primordial gonad. In YTK larvae at 3, 5, 7 and 10 days post hatch (DPH), PGCs were not yet enclosed by somatic cells, indicating the primordial gonad had not yet started to form. While at 15, 18 and 20 DPH PGCs had already settled at the genital ridge and started to become enclosed by somatic cells indicating the primordial gonad had started to develop. A higher number of PGCs were observed in the larvae at 15 and 18 DPH indicating PGCs proliferation, which corresponds with them becoming enclosed by the somatic cells. Directional migration of PGCs toward the genital ridge is a critical event in the subsequent development of a gonad. In zebrafish, mouse and chicken, stromal-cell derived factor (SDF1) signalling is one of the key molecules for PGC migration. We subsequently isolated from YTK the SDF1 (Slal-SDF1) gene, which encodes for a 98-residue precursor protein with a signal peptide at the N-terminus. There is spatial conservation between fish species of four cysteine residues at positions C9, C11, C34 and C49, expected to form disulphide bonds and stabilize the SDF structure. In YTK, Slal-SDF1 gene expression analyses shows that this gene is expressed in larvae from 1 to 22 DPH and

  19. Target-cell-derived tRNA-like primers for reverse transcription support retroviral infection at low efficiency

    DEFF Research Database (Denmark)

    Schmitz, Alexander; Lund, Anders H; Hansen, Anette C;

    2002-01-01

    Reverse transcription of a retroviral genome takes place in the cytoplasm of an infected cell by a process primed by a producer-cell-derived tRNA annealed to an 18-nucleotide primer-binding site (PBS). By an assay involving primer complementation of PBS-mutated vectors we analyzed whether t......RNA primers derived from the target cell can sustain reverse transcription during murine leukemia virus (MLV) infection. Transduction efficiencies were 4-5 orders of magnitude below those of comparable producer-cell complementations. However, successful usage of a target-cell-derived tRNA primer was proven by...... cases of correction of single mismatches between Akv-MLV vectors and complementary tRNA primers toward the primer sequence in the integrated vector. Thus, target-cell-derived tRNA-like primers are able to initiate first-strand cDNA synthesis and plus-strand transfer leading to a complete provirus...

  20. Prokaryotic Expression of Melittin/hIL-2 Mutant Fusion Protein and Its Effect on Growth of Host E. Coli%蜂毒肽/变构hIL-2融合蛋白的原核表达及其对宿主菌生长的影响

    Institute of Scientific and Technical Information of China (English)

    郑旭; 李鹏; 王斌; 鲁晓晴; 宋卫青; 赵巍; 闫志勇; 钱冬萌; 丁守怡; 宋旭霞; 徐莉莉

    2009-01-01

    目的 构建蜂毒肽(Melittin)与变构hIL-2融合基因原核表达质粒,并检测表达的融合蛋白对宿主菌E. coli生长的影响.方法 以质粒pGEX-4T-2/Melittin-IL-2(88Arg)为模板,通过PCR定点诱变为Melittin-IL-2(88Arg,125Ala).将PCR产物和pET-15b载体分别经双酶切后连接,构建重组表达质粒pET-15b/M-IL-2(88Arg,125SAla),转化E. coli BL21(DE3),IPTG诱导表达.SDS-PAGE及ELISA分析表达产物;检测诱导不同时间重组菌的A600值,绘制生长曲线,并进行活菌计数.结果 PCR扩增的目的 片段长542 bp,重组表达质粒测序分析证明目的 基因如预期突变;ELISA可检测到目的 蛋白表达,但表达量较低;SDS-PAGE分析未见目的条带;诱导表达4 h,重组菌A600值由0.8 下降至0.6;诱导2 h,活菌计数由108个/ml降至104个/ml.结论 已成功构建了原核表达质粒pET-15b/M-IL-2(88Arg,125Ala),表达的融合蛋白可能对宿主菌具有毒性,从而杀伤宿主菌.

  1. Changes in Levels of Serum IL-2 and IL-10 and Their Significance in Neonates with Unconjugated Hyperbilirubinemia Treated with Different Phototherapy Methods%不同光疗方式时新生儿高间接胆红素血症血清IL-2、IL-10水平的变化及其意义

    Institute of Scientific and Technical Information of China (English)

    黄桂兰; 陈晓

    2011-01-01

    目的 探讨不同光疗方式时新生儿高间接胆红素血症血清IL-2、IL-10水平的变化及其意义.方法 选择48例足月高间接胆红素血症新生儿为研究对象,按序贯试验分为持续光疗组24例(光疗24 h停24 h)、间断光疗组24例(光疗12h停12h×2 d),均于光疗前、光疗后24 h和48 h分别检测血清总胆红素、间接胆红素浓度,采用酶联免疫分析(ELISA)法检测血清IL-2、IL-10水平.结果 1)2组光疗后血清总胆红素、间接胆红素浓度均明显降低;光疗后24 h持续光疗组血清总胆红素、间接胆红素浓度明显低于间断光疗组(P<0.01),光疗后48 h血清总胆红素、间接胆红素浓度2组间比较差异无统计学意义(P>0.05).2)2组光疗后血清IL-2水平均明显上升;光疗后24 h持续光疗组血清IL-2水平明显高于间断光疗组(P<0.01),光疗后48 h血清IL-2水平2组间比较差异无统计学意义(P>0.05).3)持续光疗组光疗后24、48 h血清IL-10水平均较光疗前上升(P<0.001),光疗后48 h与24 h比较差异无统计学意义(P>0.05);间断光疗组光疗前后血清IL-10水平无明显变化(P>0.05),光疗后24、48 h持续光疗组血清IL-10水平均显著高于间断光疗组(P<0.001).4)血清IL-2水平与血清总胆红素、间接胆红素浓度均呈负相关(r=-0.966,r=-0.959;均P<0.01).结论 持续光疗与间断光疗降低血清胆红素浓度的疗效相同;2种不同光疗方式时高间接胆红素血症新生儿血清IL-2水平均升高,但持续光疗时血清IL-10水平升高,而间断光疗时血清IL-10水平无明显变化,提示光疗时会影响高间接胆红素血症新生儿的免疫功能;对于光疗时间较长的新生儿,同时给予适量免疫调节剂,有利于增强机体免疫力,促进疾病恢复.%Objective To explore the changes in levels of serum IL-2 and IL-10 and their significance in neonates with unconjugated hyperbilirubinemia treated with different phototherapy

  2. Use of human stem cell derived cardiomyocytes to examine sunitinib mediated cardiotoxicity and electrophysiological alterations

    Energy Technology Data Exchange (ETDEWEB)

    Cohen, J.D., E-mail: jennifer.cohen@roche.com [Early and Investigative Safety, Nonclinical Safety, Hoffmann-La Roche, 340 Kingsland Street, Nutley, NJ 07110 (United States); Babiarz, J.E., E-mail: joshua.babiarz@roche.com [Early and Investigative Safety, Nonclinical Safety, Hoffmann-La Roche, 340 Kingsland Street, Nutley, NJ 07110 (United States); Abrams, R.M., E-mail: rory.abrams@roche.com [Early and Investigative Safety, Nonclinical Safety, Hoffmann-La Roche, 340 Kingsland Street, Nutley, NJ 07110 (United States); Guo, L., E-mail: liang.guo@roche.com [Early and Investigative Safety, Nonclinical Safety, Hoffmann-La Roche, 340 Kingsland Street, Nutley, NJ 07110 (United States); Kameoka, S., E-mail: sei.kameoka@roche.com [Early and Investigative Safety, Nonclinical Safety, Hoffmann-La Roche, 340 Kingsland Street, Nutley, NJ 07110 (United States); Chiao, E., E-mail: eric.chiao@roche.com [Early and Investigative Safety, Nonclinical Safety, Hoffmann-La Roche, 340 Kingsland Street, Nutley, NJ 07110 (United States); Taunton, J., E-mail: taunton@cmp.ucsf.edu [Howard Hughes Medical Institute, Cellular and Molecular Pharmacology, University California San Francisco, San Francisco, CA 94158 (United States); Kolaja, K.L., E-mail: kyle.kolaja@roche.com [Early and Investigative Safety, Nonclinical Safety, Hoffmann-La Roche, 340 Kingsland Street, Nutley, NJ 07110 (United States)

    2011-11-15

    Sunitinib, an oral tyrosine kinase inhibitor approved to treat advanced renal cell carcinoma and gastrointestinal stroma tumor, is associated with clinical cardiac toxicity. Although the precise mechanism of sunitinib cardiotoxicity is not known, both the key metabolic energy regulator, AMP-activated protein kinase (AMPK), and ribosomal S 6 kinase (RSK) have been hypothesized as causative, albeit based on rodent models. To study the mechanism of sunitinib-mediated cardiotoxicity in a human model, induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) having electrophysiological and contractile properties of native cardiac tissue were investigated. Sunitinib was cardiotoxic in a dose-dependent manner with an IC{sub 50} in the low micromolar range, observed by a loss of cellular ATP, an increase in oxidized glutathione, and induction of apoptosis in iPSC-CMs. Pretreatment of iPSC-CMs with AMPK activators AICAR or metformin, increased the phosphorylation of pAMPK-T172 and pACC-S79, but only marginally attenuated sunitinib mediated cell death. Furthermore, additional inhibitors of AMPK were not directly cytotoxic to iPSC-CMs up to 250 {mu}M concentrations. Inhibition of RSK with a highly specific, irreversible, small molecule inhibitor (RSK-FMK-MEA) did not induce cytotoxicity in iPSC-CMs below 250 {mu}M. Extensive electrophysiological analysis of sunitinib and RSK-FMK-MEA mediated conduction effects were performed. Taken together, these findings suggest that inhibition of AMPK and RSK are not a major component of sunitinib-induced cardiotoxicity. Although the exact mechanism of cardiotoxicity of sunitinib is not known, it is likely due to inhibition of multiple kinases simultaneously. These data highlight the utility of human iPSC-CMs in investigating the potential molecular mechanisms underlying drug-induced cardiotoxicity. -- Highlights: Black-Right-Pointing-Pointer Cytoxic effect of sunitinib on human stem cell derived cardiomyocytes Black

  3. Neuroantigen-Specific CD4 Cells Expressing Interferon-γ (IFN-γ), Interleukin (IL)-2 and IL-3 in a Mutually Exclusive Manner Prevail in Experimental Allergic Encephalomyelitis (EAE).

    Science.gov (United States)

    Karulin, Alexey Y; Quast, Stefan; Hesse, Maike D; Lehmann, Paul V

    2012-08-24

    Experimental allergic encephalomyelitis (EAE) is mediated by neuroantigen-specific pro-inflammatory T cells of the Th1 and Th17 effector class. Th-17 cells can be clearly defined by expression of IL-17, but not IFN-γ, IL-2 or IL-3. Th1 cells do not express IL-17, but it is unclear presently to what extent they co-express the cytokines canonically assigned to Th1 immunity (i.e., IFN-γ, IL-2 and IL-3) and whether CD4 cells producing these cytokines indeed belong to a single Th1 lineage. It is also unclear to what extent the Th1 response in EAE entails polyfunctional T cells that co-express IFN-γ and IL-2. Therefore, we dissected the Th1 cytokine signature of neuroantigen-specific CD4 cells studying at single cell resolution co-expression of IFN-γ, IL-2 and IL-3 using dual color cytokine ELISPOT analysis. Shortly after immunization, in the draining lymph nodes (dLN), the overall cytokine signature of the neuroantigen-specific CD4 cells was highly type 1-polarized, but IFN-γ, IL-2, and IL-3 were each secreted by different CD4 cells in a mutually exclusive manner. This single cell - single cytokine profile was stable through the course of chronic EAE-polyfunctional CD4 cells co-expressing IL-2 and IFN-γ presented less than 5% of the neuroantigen-specific T cells, even in the inflamed CNS itself. The neuroantigen-specific CD4 cells that expressed IFN-γ, IL-2 and IL-3 in a mutually exclusive manner exhibited similar functional avidities and kinetics of cytokine production, but showed different tissue distributions. These data suggest that Th1 cells do not belong to a single lineage, but different Th1 subpopulations jointly mediate Th1 immunity.

  4. Higher susceptibility of NOD/LtSz-scid Il2rg−/− NSG mice to xenotransplanted lung cancer cell lines

    International Nuclear Information System (INIS)

    No lung cancer xenograft model using non-obese diabetic (NOD)-scid Il2rg−/− mice has been reported. The purpose of this study is to select a suitable mouse strain as a xenogenic host for testing tumorigenicity of lung cancer. We directly compared the susceptibility of four immunodeficient mouse strains, c-nu, C.B-17 scid, NOD-scid, and NOD/LtSz-scid Il2rg−/− (NSG) mice, for tumor formation from xenotransplanted lung cancer cell lines. Various numbers (101–105 cells/head) of two lung cancer cell lines, A549 and EBC1, were subcutaneously inoculated and tumor sizes were measured every week up to 12 weeks. When 104 EBC1 cells were inoculated, no tumor formation was observed in BALB/c-nu or C.B-17 scid mice. Tumors developed in two of the five NOD-scid mice (40%) and in all the five NSG mice (100%). When 103 EBC1 cells were injected, no tumors developed in any strain other than NSG mice, while tumorigenesis was achieved in all the five NSG mice (100%, P=0.0079) within 9 weeks. NSG mice similarly showed higher susceptibility to xenotransplantation of A549 cells. Tumor formation was observed only in NSG mice after inoculation of 103 or fewer A549 cells (40% vs 0% in 15 NSG mice compared with others, respectively, P=0.0169). We confirmed that the engrafted tumors originated from inoculated human lung cancer cells by immunohistochemical staining with human cytokeratin and vimentin. NSG mice may be the most suitable strain for testing tumorigenicity of lung cancer, especially if only a few cells are available

  5. Three ulcerative colitis susceptibility loci are associated with primary sclerosing cholangitis and indicate a role for IL2, REL and CARD9

    Science.gov (United States)

    Janse, Marcel; Lamberts, Laetitia E; Franke, Lude; Raychaudhuri, Soumya; Ellinghaus, Eva; MuriBoberg, Kirsten; Melum, Espen; Folseraas, Trine; Schrumpf, Erik; Bergquist, Annika; Bjornsson, Einar; Fu, Jingyuan; Westra, Harm Jan; Groen, Harry JM; Fehrmann, Rudolf SN; Smolonska, Joanna; van den Berg, Leonard H; Ophoff, Roel A; Porte, Robert J; Weismuller, Tobias J; Wedemeyer, Jochen; Schramm, Christoph; Sterneck, Martina; Gunther, Rainer; Braun, Felix; Vermeire, Severine; Henckaerts, Liesbet; Wijmenga, Cisca; Ponsioen, Cyriel Y.; Schreiber, Stefan; HKarlsen, Tom; Franke, Andre; Weersma, Rinse K

    2011-01-01

    Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease characterized by inflammation and fibrosis of the bile ducts. Both environmental and genetic factors contribute to its pathogenesis. To further clarify its genetic background, we investigated susceptibility loci recently identified for ulcerative colitis (UC) in a large cohort of 1186PSC patients and 1748 controls. Single nucleotide polymorphisms (SNPs) tagging 13 UC susceptibility loci were initially genotyped in 854 PSC patients and 1491 controls from the Benelux (331 cases, 735 controls), Germany (265 cases, 368 controls) and Scandinavia (258 cases, 388 controls). Subsequently, a joint analysis was performed with an independent second Scandinavian cohort (332 cases, 257 controls). SNPs at chromosomes2p16 (p value 4.12×10−4), 4q27 (p value 4.10×10−5) and 9q34 (p value 8.41×10−4) were associated with PSC in the joint analysis after correcting for multiple testing. In PSC patients without inflammatory bowel disease(IBD), SNPs at 4q27and9q34 were nominally associated (p<0.05). We applied additional in silico analyses to identify likely candidate genes at PSC susceptibility loci. To identify non-random, evidence-based links we used GRAIL analysis showing interconnectivity between genes in six out of in total nine PSC-associated regions. Expression quantitative trait analysis from 1469 Dutch and UK individuals demonstrated that five out of nine SNPs had an effect on cis-gene expression. These analyses prioritized IL2, CARD9 and REL as novel candidates. Conclusion We have identified three UC susceptibility loci to be associated with PSC, harboring the putative candidate genes REL, IL2 and CARD9. These results add to the scarce knowledge on the genetic background of PSC and imply an important role for both innate and adaptive immunological factors. PMID:21425313

  6. Potential Therapies by Stem Cell-Derived Exosomes in CNS Diseases: Focusing on the Neurogenic Niche

    Directory of Open Access Journals (Sweden)

    Alejandro Luarte

    2016-01-01

    Full Text Available Neurodegenerative disorders are one of the leading causes of death and disability and one of the biggest burdens on health care systems. Novel approaches using various types of stem cells have been proposed to treat common neurodegenerative disorders such as Alzheimer’s Disease, Parkinson’s Disease, or stroke. Moreover, as the secretome of these cells appears to be of greater benefit compared to the cells themselves, the extracellular components responsible for its therapeutic benefit have been explored. Stem cells, as well as most cells, release extracellular vesicles such as exosomes, which are nanovesicles able to target specific cell types and thus to modify their function by delivering proteins, lipids, and nucleic acids. Exosomes have recently been tested in vivo and in vitro as therapeutic conveyors for the treatment of diseases. As such, they could be engineered to target specific populations of cells within the CNS. Considering the fact that many degenerative brain diseases have an impact on adult neurogenesis, we discuss how the modulation of the adult neurogenic niches may be a therapeutic target of stem cell-derived exosomes. These novel approaches should be examined in cellular and animal models to provide better, more effective, and specific therapeutic tools in the future.

  7. Potential Therapies by Stem Cell-Derived Exosomes in CNS Diseases: Focusing on the Neurogenic Niche

    Science.gov (United States)

    Luarte, Alejandro; Bátiz, Luis Federico; Wyneken, Ursula; Lafourcade, Carlos

    2016-01-01

    Neurodegenerative disorders are one of the leading causes of death and disability and one of the biggest burdens on health care systems. Novel approaches using various types of stem cells have been proposed to treat common neurodegenerative disorders such as Alzheimer's Disease, Parkinson's Disease, or stroke. Moreover, as the secretome of these cells appears to be of greater benefit compared to the cells themselves, the extracellular components responsible for its therapeutic benefit have been explored. Stem cells, as well as most cells, release extracellular vesicles such as exosomes, which are nanovesicles able to target specific cell types and thus to modify their function by delivering proteins, lipids, and nucleic acids. Exosomes have recently been tested in vivo and in vitro as therapeutic conveyors for the treatment of diseases. As such, they could be engineered to target specific populations of cells within the CNS. Considering the fact that many degenerative brain diseases have an impact on adult neurogenesis, we discuss how the modulation of the adult neurogenic niches may be a therapeutic target of stem cell-derived exosomes. These novel approaches should be examined in cellular and animal models to provide better, more effective, and specific therapeutic tools in the future. PMID:27195011

  8. Importance of being Nernst: Synaptic activity andfunctional relevance in stem cell-derived neurons

    Institute of Scientific and Technical Information of China (English)

    2015-01-01

    Functional synaptogenesis and network emergence aresignature endpoints of neurogenesis. These behaviorsprovide higher-order confirmation that biochemicaland cellular processes necessary for neurotransmitterrelease, post-synaptic detection and network propagation of neuronal activity have been properly expressed andcoordinated among cells. The development of synapticneurotransmission can therefore be considered a definingproperty of neurons. Although dissociated primaryneuron cultures readily form functioning synapsesand network behaviors in vitro , continuously culturedneurogenic cell lines have historically failed to meet thesecriteria. Therefore, in vitro -derived neuron models thatdevelop synaptic transmission are critically needed for awide array of studies, including molecular neuroscience,developmental neurogenesis, disease research andneurotoxicology. Over the last decade, neurons derivedfrom various stem cell lines have shown varying ability todevelop into functionally mature neurons. In this review,we will discuss the neurogenic potential of various stemcells populations, addressing strengths and weaknessesof each, with particular attention to the emergenceof functional behaviors. We will propose methods tofunctionally characterize new stem cell-derived neuron(SCN) platforms to improve their reliability as physiologicalrelevant models. Finally, we will review howsynaptically active SCNs can be applied to accelerateresearch in a variety of areas. Ultimately, emphasizingthe critical importance of synaptic activity and networkresponses as a marker of neuronal maturation is anticipatedto result in in vitro findings that better translateto efficacious clinical treatments.

  9. Appearance of differentiated cells derived from polar body nuclei in the silkworm, Bombyx mori

    Directory of Open Access Journals (Sweden)

    Hiroki eSakai

    2013-09-01

    Full Text Available AbstractIn Bombyx mori, polar body nuclei are observed until 9h after egg lying, however, the fate of polar body nuclei remains unclear. To examine the fate of polar body nuclei, we employed a mutation of serosal cell pigmentation, pink-eyed white egg (pe. The heterozygous pe/+pe females produced black serosal cells in white eggs, while pe/pe females did not produce black serosal cells in white eggs. These results suggest that the appearance of black serosal cells in white eggs depends on the genotype (pe/ +pe of the mother. Because the polar body nuclei had +pe genes in the white eggs laid by a pe/ +pe female, polar body nuclei participate in development and differentiate into functional cell (serosal cells. Analyses of serosal cells pigmentation indicated that approximately 30% of the eggs contained polar-body-nucleus-derived cells. These results demonstrate that polar-body-nucleus-derived cells appeared at a high frequency under natural conditions. Approximately 80% of polar-body-nucleus-derived cells appeared near the anterior pole and the dorsal side, which is opposite to where embryogenesis occurs. The number of cells derived from the polar body nuclei was very low. Approximately 26 % of these eggs contained only one black serosal cell. PCR-based analysis revealed that the polar-body-nucleus-derived cells disappeared in late embryonic stages (stage 25. Overall, polar-body-nuclei-derived cells were unlikely to contribute to embryos.

  10. Human pluripotent stem cell-derived neural constructs for predicting neural toxicity.

    Science.gov (United States)

    Schwartz, Michael P; Hou, Zhonggang; Propson, Nicholas E; Zhang, Jue; Engstrom, Collin J; Santos Costa, Vitor; Jiang, Peng; Nguyen, Bao Kim; Bolin, Jennifer M; Daly, William; Wang, Yu; Stewart, Ron; Page, C David; Murphy, William L; Thomson, James A

    2015-10-01

    Human pluripotent stem cell-based in vitro models that reflect human physiology have the potential to reduce the number of drug failures in clinical trials and offer a cost-effective approach for assessing chemical safety. Here, human embryonic stem (ES) cell-derived neural progenitor cells, endothelial cells, mesenchymal stem cells, and microglia/macrophage precursors were combined on chemically defined polyethylene glycol hydrogels and cultured in serum-free medium to model cellular interactions within the developing brain. The precursors self-assembled into 3D neural constructs with diverse neuronal and glial populations, interconnected vascular networks, and ramified microglia. Replicate constructs were reproducible by RNA sequencing (RNA-Seq) and expressed neurogenesis, vasculature development, and microglia genes. Linear support vector machines were used to construct a predictive model from RNA-Seq data for 240 neural constructs treated with 34 toxic and 26 nontoxic chemicals. The predictive model was evaluated using two standard hold-out testing methods: a nearly unbiased leave-one-out cross-validation for the 60 training compounds and an unbiased blinded trial using a single hold-out set of 10 additional chemicals. The linear support vector produced an estimate for future data of 0.91 in the cross-validation experiment and correctly classified 9 of 10 chemicals in the blinded trial.

  11. Reversal of diabetes with insulin-producing cells derived in vitro from human pluripotent stem cells.

    Science.gov (United States)

    Rezania, Alireza; Bruin, Jennifer E; Arora, Payal; Rubin, Allison; Batushansky, Irina; Asadi, Ali; O'Dwyer, Shannon; Quiskamp, Nina; Mojibian, Majid; Albrecht, Tobias; Yang, Yu Hsuan Carol; Johnson, James D; Kieffer, Timothy J

    2014-11-01

    Transplantation of pancreatic progenitors or insulin-secreting cells derived from human embryonic stem cells (hESCs) has been proposed as a therapy for diabetes. We describe a seven-stage protocol that efficiently converts hESCs into insulin-producing cells. Stage (S) 7 cells expressed key markers of mature pancreatic beta cells, including MAFA, and displayed glucose-stimulated insulin secretion similar to that of human islets during static incubations in vitro. Additional characterization using single-cell imaging and dynamic glucose stimulation assays revealed similarities but also notable differences between S7 insulin-secreting cells and primary human beta cells. Nevertheless, S7 cells rapidly reversed diabetes in mice within 40 days, roughly four times faster than pancreatic progenitors. Therefore, although S7 cells are not fully equivalent to mature beta cells, their capacity for glucose-responsive insulin secretion and rapid reversal of diabetes in vivo makes them a promising alternative to pancreatic progenitor cells or cadaveric islets for the treatment of diabetes.

  12. Muse Cells, a New Type of Pluripotent Stem Cell Derived from Human Fibroblasts.

    Science.gov (United States)

    Liu, Qi; Zhang, Ru-Zhi; Li, Di; Cheng, Sai; Yang, Yu-Hua; Tian, Ting; Pan, Xiao-Ru

    2016-04-01

    A new type of mesenchymal stem cells (MSCs) that expresses stage-specific embryonic antigen 3 (SSEA-3) and the mesenchymal cell marker CD105 are known as multilineage-differentiating stress-enduring (Muse) cells. Studies have shown that stem cells in suspension cultures are more likely to generate embryoid body-like stem cell spheres and maintain an undifferentiated phenotype and pluripotency. We separated Muse cells derived from human dermal fibroblasts by long-term trypsin incubation (LTT) through suspension cultures in methylcellulose. The Muse cells obtained expressed several pluripotency markers, including Nanog, Oct4, Sox2, and SSEA-3, and could differentiate in vitro into cells of the three germ layers, such as hepatocytes (endodermal), neural cells (ectodermal) and adipocytes, and osteocytes (mesodermal cells). These cells showed a low level of DNA methylation and a high nucleo-cytoplasmic ratio. Our study provides an innovative and exciting platform for exploring the potential cell-based therapy of various human diseases using Muse cells as well as their great possibility for regenerative medicine. PMID:27055628

  13. Stem cell-derived exosomes as a therapeutic tool for cardiovascular disease

    Science.gov (United States)

    Suzuki, Etsu; Fujita, Daishi; Takahashi, Masao; Oba, Shigeyoshi; Nishimatsu, Hiroaki

    2016-01-01

    Mesenchymal stem cells (MSCs) have been used to treat patients suffering from acute myocardial infarction (AMI) and subsequent heart failure. Although it was originally assumed that MSCs differentiated into heart cells such as cardiomyocytes, recent evidence suggests that the differentiation capacity of MSCs is minimal and that injected MSCs restore cardiac function via the secretion of paracrine factors. MSCs secrete paracrine factors in not only naked forms but also membrane vesicles including exosomes containing bioactive substances such as proteins, messenger RNAs, and microRNAs. Although the details remain unclear, these bioactive molecules are selectively sorted in exosomes that are then released from donor cells in a regulated manner. Furthermore, exosomes are specifically internalized by recipient cells via ligand-receptor interactions. Thus, exosomes are promising natural vehicles that stably and specifically transport bioactive molecules to recipient cells. Indeed, stem cell-derived exosomes have been successfully used to treat cardiovascular disease (CVD), such as AMI, stroke, and pulmonary hypertension, in animal models, and their efficacy has been demonstrated. Therefore, exosome administration may be a promising strategy for the treatment of CVD. Furthermore, modifications of exosomal contents may enhance their therapeutic effects. Future clinical studies are required to confirm the efficacy of exosome treatment for CVD. PMID:27679686

  14. Stretch Injury of Human Induced Pluripotent Stem Cell Derived Neurons in a 96 Well Format

    Science.gov (United States)

    Sherman, Sydney A.; Phillips, Jack K.; Costa, J. Tighe; Cho, Frances S.; Oungoulian, Sevan R.; Finan, John D.

    2016-01-01

    Traumatic brain injury (TBI) is a major cause of mortality and morbidity with limited therapeutic options. Traumatic axonal injury (TAI) is an important component of TBI pathology. It is difficult to reproduce TAI in animal models of closed head injury, but in vitro stretch injury models reproduce clinical TAI pathology. Existing in vitro models employ primary rodent neurons or human cancer cell line cells in low throughput formats. This in vitro neuronal stretch injury model employs human induced pluripotent stem cell-derived neurons (hiPSCNs) in a 96 well format. Silicone membranes were attached to 96 well plate tops to create stretchable, culture substrates. A custom-built device was designed and validated to apply repeatable, biofidelic strains and strain rates to these plates. A high content approach was used to measure injury in a hypothesis-free manner. These measurements are shown to provide a sensitive, dose-dependent, multi-modal description of the response to mechanical insult. hiPSCNs transition from healthy to injured phenotype at approximately 35% Lagrangian strain. Continued development of this model may create novel opportunities for drug discovery and exploration of the role of human genotype in TAI pathology. PMID:27671211

  15. Puerarin Facilitates T-Tubule Development of Murine Embryonic Stem Cell-Derived Cardiomyocytes

    Directory of Open Access Journals (Sweden)

    Lu Wang

    2014-07-01

    Full Text Available Aims: The embryonic stem cell-derived cardiomyocytes (ES-CM is one of the promising cell sources for repopulation of damaged myocardium. However, ES-CMs present immature structure, which impairs their integration with host tissue and functional regeneration. This study used murine ES-CMs as an in vitro model of cardiomyogenesis to elucidate the effect of puerarin, the main compound found in the traditional Chinese medicine the herb Radix puerariae, on t-tubule development of murine ES-CMs. Methods: Electron microscope was employed to examine the ultrastructure. The investigation of transverse-tubules (t-tubules was performed by Di-8-ANEPPS staining. Quantitative real-time PCR was utilized to study the transcript level of genes related to t-tubule development. Results: We found that long-term application of puerarin throughout cardiac differentiation improved myofibril array and sarcomeres formation, and significantly facilitated t-tubules development of ES-CMs. The transcript levels of caveolin-3, amphiphysin-2 and junctophinlin-2, which are crucial for the formation and development of t-tubules, were significantly upregulated by puerarin treatment. Furthermore, puerarin repressed the expression of miR-22, which targets to caveolin-3. Conclusion: Our data showed that puerarin facilitates t-tubule development of murine ES-CMs. This might be related to the repression of miR-22 by puerarin and upregulation of Cav3, Bin1 and JP2 transcripts.

  16. "Kill" the messenger: Targeting of cell-derived microparticles in lupus nephritis.

    Science.gov (United States)

    Nielsen, Christoffer T; Rasmussen, Niclas S; Heegaard, Niels H H; Jacobsen, Søren

    2016-07-01

    Immune complex (IC) deposition in the glomerular basement membrane (GBM) is a key early pathogenic event in lupus nephritis (LN). The clarification of the mechanisms behind IC deposition will enable targeted therapy in the future. Circulating cell-derived microparticles (MPs) have been proposed as major sources of extracellular autoantigens and ICs and triggers of autoimmunity in LN. The overabundance of galectin-3-binding protein (G3BP) along with immunoglobulins and a few other proteins specifically distinguish circulating MPs in patients with systemic lupus erythematosus (SLE), and this is most pronounced in patients with active LN. G3BP co-localizes with deposited ICs in renal biopsies from LN patients supporting a significant presence of MPs in the IC deposits. G3BP binds strongly to glomerular basement membrane proteins and integrins. Accordingly, MP surface proteins, especially G3BP, may be essential for the deposition of ICs in kidneys and thus for the ensuing formation of MP-derived electron dense structures in the GBM, and immune activation in LN. This review focuses on the notion of targeting surface molecules on MPs as an entirely novel treatment strategy in LN. By targeting MPs, a double hit may be achieved by attenuating both the autoantigenic fueling of immune complexes and the triggering of the adaptive immune system. Thereby, early pathogenic events may be blocked in contrast to current treatment strategies that primarily target and modulate later events in the cellular and humoral immune response.

  17. Engineered Microenvironments for the Maturation and Observation of Human Embryonic Stem Cell Derived Cardiomyocytes

    Science.gov (United States)

    Salick, Max R.

    The human heart is a dynamic system that undergoes substantial changes as it develops and adapts to the body's growing needs. To better understand the physiology of the heart, researchers have begun to produce immature heart muscle cells, or cardiomyocytes, from pluripotent stem cell sources with remarkable efficiency. These stem cell-derived cardiomyocytes hold great potential in the understanding and treatment of heart disease; however, even after prolonged culture, these cells continue to exhibit an immature phenotype, as indicated by poor sarcomere organization and calcium handling, among other features. The lack of maturation that is observed in these cardiomyocytes greatly limits their applicability towards drug screening, disease modeling, and cell therapy applications. The mechanical environment surrounding a cell has been repeatedly shown to have a large impact on that cell's behavior. For this reason, we have implemented micropatterning methods to mimic the level of alignment that occurs in the heart in vivo in order to study how this alignment may help the cells to produce a more mature sarcomere phenotype. It was discovered that the level of sarcomere organization of a cardiomyocyte can be strongly influenced by the micropattern lane geometry on which it adheres. Steps were taken to optimize this micropattern platform, and studies of protein organization, gene expression, and myofibrillogenesis were conducted. Additionally, a set of programs was developed to provide quantitative analysis of the level of sarcomere organization, as well as to assist with several other tissue engineering applications.

  18. The similarity between human embryonic stem cell-derived epithelial cells and ameloblast-lineage cells

    Institute of Scientific and Technical Information of China (English)

    Li-Wei Zheng; Logan Linthicum; Pamela K DenBesten; Yan Zhang

    2013-01-01

    This study aimed to compare epithelial cells derived from human embryonic stem cells (hESCs) to human ameloblast-lineage cells (ALCs), as a way to determine their potential use as a cell source for ameloblast regeneration. Induced by various concentrations of bone morphogenetic protein 4 (BMP4), retinoic acid (RA) and lithium chloride (LiCI) for 7 days, hESCs adopted cobble-stone epithelial phenotype (hESC-derived epithelial cells (ES-ECs)) and expressed cytokeratin 14. Compared with ALCs and oral epithelial cells (OE), ES-ECs expressed amelogenesis-associated genes similar to ALCs. ES-ECs were compared with human fetal skin epithelium, human fetal oral buccal mucosal epithelial cells and human ALCs for their expression pattern of cytokeratins as well. ALCs had relatively high expression levels of cytokeratin 76, which ,vas also found to be upregulated in ES-ECs. Based on the present study, with the similarity of gene expression with ALCs, ES-ECs are a promising potential cell source for regeneration, which are not available in erupted human teeth for regeneration of enamel.

  19. Functional differentiation of stem cell-derived neurons from different murine backgrounds

    Directory of Open Access Journals (Sweden)

    Lydia eBarth

    2014-02-01

    Full Text Available Murine stem cell derived-neurons have been used to study a wide variety of neuropsychiatric diseases with a hereditary component, ranging from autism to Alzheimer’s. While a significant amount of data on their molecular biology has been generated, there is little data on the physiology of these cultures. Different mouse strains show clear differences in behavioural and other neurobiologically relevant readouts. We have studied the physiology of early differentiation and network formation in neuronal cultures derived from three different mouse embryonic stem cell lines. We have found largely overlapping patterns with some significant differences in the timing of the functional milestones. Neurons from R1 showed the fastest development of intrinsic excitability, while E14Tg2a and J1 were slower. This was also reflected in an earlier appearance of synaptic activity in R1 cultures, while E14Tg2a and J1 were delayed by up to two days. In conclusion, stem cells from all backgrounds could be successfully differentiated into functioning neural networks with similar developmental patterns. Differences in the timing of specific milestones, suggest that control cell lines and time-points should be carefully chosen when investigating genetic alterations that lead to subtle deficits in neuronal function.

  20. Low antigenicity of hematopoietic progenitor cells derived from human ES cells

    Directory of Open Access Journals (Sweden)

    Eun-Mi Kim

    2010-02-01

    Full Text Available Eun-Mi Kim1, Nicholas Zavazava1,21Department of Internal Medicine, University of Iowa and Veterans Affairs Medical Center, Iowa City, Iowa, USA; 2Immunology Graduate Program, University of Iowa, Iowa City, Iowa, USAAbstract: Human embryonic stem (hES cells are essential for improved understanding of diseases and our ability to probe new therapies for use in humans. Currently, bone marrow cells and cord blood cells are used for transplantation into patients with hematopoietic malignancies, immunodeficiencies and in some cases for the treatment of autoimmune diseases. However, due to the high immunogenicity of these hematopoietic cells, toxic regimens of drugs are required for preconditioning and prevention of rejection. Here, we investigated the efficiency of deriving hematopoietic progenitor cells (HPCs from the hES cell line H13, after co-culturing with the murine stromal cell line OP9. We show that HPCs derived from the H13 ES cells poorly express major histocompatibility complex (MHC class I and no detectable class II antigens (HLA-DR. These characteristics make hES cell-derived hematopoietic cells (HPCs ideal candidates for transplantation across MHC barriers under minimal immunosuppression.Keywords: human embryonic stem cells, H13, hematopoiesis, OP9 stromal cells, immunogenicity

  1. Process Extension from Embryonic Stem Cell-Derived Motor Neurons through Synthetic Extracellular Matrix Mimics

    Science.gov (United States)

    McKinnon, Daniel Devaud

    This thesis focuses on studying the extension of motor axons through synthetic poly(ethylene glycol) PEG hydrogels that have been modified with biochemical functionalities to render them more biologically relevant. Specifically, the research strategy is to encapsulate embryonic stem cell-derived motor neurons (ESMNs) in synthetic PEG hydrogels crosslinked through three different chemistries providing three mechanisms for dynamically tuning material properties. First, a covalently crosslinked, enzymatically degradable hydrogel is developed and exploited to study the biophysical dynamics of axon extension and matrix remodeling. It is demonstrated that dispersed motor neurons require a battery of adhesive peptides and growth factors to maintain viability and extend axons while those in contact with supportive neuroglial cells do not. Additionally, cell-degradable crosslinker peptides and a soft modulus mimicking that of the spinal cord are requirements for axon extension. However, because local degradation of the hydrogel results in a cellular environment significantly different than that of the bulk, enzymatically degradable peptide crosslinkers were replaced with reversible covalent hydrazone bonds to study the effect of hydrogel modulus on axon extension. This material is characterized in detail and used to measure forces involved in axon extension. Finally, a hydrogel with photocleavable linkers incorporated into the network structure is exploited to explore motor axon response to physical channels. This system is used to direct the growth of motor axons towards co-cultured myotubes, resulting in the formation of an in vitro neural circuit.

  2. Automated Electrophysiological and Pharmacological Evaluation of Human Pluripotent Stem Cell-Derived Cardiomyocytes.

    Science.gov (United States)

    Rajamohan, Divya; Kalra, Spandan; Duc Hoang, Minh; George, Vinoj; Staniforth, Andrew; Russell, Hugh; Yang, Xuebin; Denning, Chris

    2016-03-15

    Automated planar patch clamp systems are widely used in drug evaluation studies because of their ability to provide accurate, reliable, and reproducible data in a high-throughput manner. Typically, CHO and HEK tumorigenic cell lines overexpressing single ion channels are used since they can be harvested as high-density, homogenous, single-cell suspensions. While human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are physiologically more relevant, these cells are fragile, have complex culture requirements, are inherently heterogeneous, and are expensive to produce, which has restricted their use on automated patch clamp (APC) devices. Here, we used high efficiency differentiation protocols to produce cardiomyocytes from six different hPSC lines for analysis on the Patchliner (Nanion Technologies GmbH) APC platform. We developed a two-step cell preparation protocol that yielded cell catch rates and whole-cell breakthroughs of ∼80%, with ∼40% of these cells allowing electrical activity to be recorded. The protocol permitted formation of long-lasting (>15 min), high quality seals (>2 GΩ) in both voltage- and current-clamp modes. This enabled density of sodium, calcium, and potassium currents to be evaluated, along with dose-response curves to their respective channel inhibitors, tetrodotoxin, nifedipine, and E-4031. Thus, we show the feasibility of using the Patchliner platform for automated evaluation of the electrophysiology and pharmacology of hPSC-CMs, which will enable considerable increase in throughput for reliable and efficient drug evaluation. PMID:26906236

  3. Generating and characterizing the mechanical properties of cell-derived matrices using atomic force microscopy.

    Science.gov (United States)

    Tello, Marta; Spenlé, Caroline; Hemmerlé, Joseph; Mercier, Luc; Fabre, Roxane; Allio, Guillaume; Simon-Assmann, Patricia; Goetz, Jacky G

    2016-02-01

    Mechanical interaction between cells and their surrounding extracellular matrix (ECM) controls key processes such as proliferation, differentiation and motility. For many years, two-dimensional (2D) models were used to better understand the interactions between cells and their surrounding ECM. More recently, variation of the mechanical properties of tissues has been reported to play a major role in physiological and pathological scenarios such as cancer progression. The 3D architecture of the ECM finely tunes cellular behavior to perform physiologically relevant tasks. Technical limitations prevented scientists from obtaining accurate assessment of the mechanical properties of physiologically realistic matrices. There is therefore a need for combining the production of high-quality cell-derived 3D matrices (CDMs) and the characterization of their topographical and mechanical properties. Here, we describe methods that allow to accurately measure the young modulus of matrices produced by various cellular types. In the first part, we will describe and review several protocols for generating CDMs matrices from endothelial, epithelial, fibroblastic, muscle and mesenchymal stem cells. We will discuss tools allowing the characterization of the topographical details as well as of the protein content of such CDMs. In a second part, we will report the methodologies that can be used, based on atomic force microscopy, to accurately evaluate the stiffness properties of the CDMs through the quantification of their young modulus. Altogether, such methodologies allow characterizing the stiffness and topography of matrices deposited by the cells, which is key for the understanding of cellular behavior in physiological conditions.

  4. Mast cell-derived neurotrophin 4 mediates allergen-induced airway hyperinnervation in early life

    Science.gov (United States)

    Patel, Kruti R.; Aven, Linh; Shao, Fengzhi; Krishnamoorthy, Nandini; Duvall, Melody G.; Levy, Bruce D.; Ai, Xingbin

    2016-01-01

    Asthma often progresses from early episodes of insults. How early life events connect to long-term airway dysfunction remains poorly understood. We demonstrated previously that increased neurotrophin 4 (NT4) levels following early life allergen exposure cause persistent changes in airway smooth muscle (ASM) innervation and airway hyper-reactivity (AHR) in mice. Herein, we identify pulmonary mast cells as a key source of aberrant NT4 expression following early insults. NT4 is selectively expressed by ASM and mast cells in mice, nonhuman primates and humans. We show in mice that mast cell-derived NT4 is dispensable for ASM innervation during development. However, upon insults, mast cells expand in number and degranulate to release NT4 and thus become the major source of NT4 under pathological condition. Adoptive transfer of wild type mast cells, but not NT4−/− mast cells restores ASM hyperinnervation and AHR in KitW-sh/W-sh mice following early life insults. Notably, an infant nonhuman primate model of asthma also exhibits ASM hyperinnervation associated with the expansion and degranulation of mast cells. Together, these findings identify an essential role of mast cells in mediating ASM hyperinnervation following early life insults by producing NT4. This role may be evolutionarily conserved in linking early insults to long-term airway dysfunction. PMID:26860818

  5. Stem cell-derived exosomes as a therapeutic tool for cardiovascular disease

    Science.gov (United States)

    Suzuki, Etsu; Fujita, Daishi; Takahashi, Masao; Oba, Shigeyoshi; Nishimatsu, Hiroaki

    2016-01-01

    Mesenchymal stem cells (MSCs) have been used to treat patients suffering from acute myocardial infarction (AMI) and subsequent heart failure. Although it was originally assumed that MSCs differentiated into heart cells such as cardiomyocytes, recent evidence suggests that the differentiation capacity of MSCs is minimal and that injected MSCs restore cardiac function via the secretion of paracrine factors. MSCs secrete paracrine factors in not only naked forms but also membrane vesicles including exosomes containing bioactive substances such as proteins, messenger RNAs, and microRNAs. Although the details remain unclear, these bioactive molecules are selectively sorted in exosomes that are then released from donor cells in a regulated manner. Furthermore, exosomes are specifically internalized by recipient cells via ligand-receptor interactions. Thus, exosomes are promising natural vehicles that stably and specifically transport bioactive molecules to recipient cells. Indeed, stem cell-derived exosomes have been successfully used to treat cardiovascular disease (CVD), such as AMI, stroke, and pulmonary hypertension, in animal models, and their efficacy has been demonstrated. Therefore, exosome administration may be a promising strategy for the treatment of CVD. Furthermore, modifications of exosomal contents may enhance their therapeutic effects. Future clinical studies are required to confirm the efficacy of exosome treatment for CVD.

  6. Human embryonic stem cell-derived neuronal cells form spontaneously active neuronal networks in vitro.

    Science.gov (United States)

    Heikkilä, Teemu J; Ylä-Outinen, Laura; Tanskanen, Jarno M A; Lappalainen, Riikka S; Skottman, Heli; Suuronen, Riitta; Mikkonen, Jarno E; Hyttinen, Jari A K; Narkilahti, Susanna

    2009-07-01

    The production of functional human embryonic stem cell (hESC)-derived neuronal cells is critical for the application of hESCs in treating neurodegenerative disorders. To study the potential functionality of hESC-derived neurons, we cultured and monitored the development of hESC-derived neuronal networks on microelectrode arrays. Immunocytochemical studies revealed that these networks were positive for the neuronal marker proteins beta-tubulin(III) and microtubule-associated protein 2 (MAP-2). The hESC-derived neuronal networks were spontaneously active and exhibited a multitude of electrical impulse firing patterns. Synchronous bursts of electrical activity similar to those reported for hippocampal neurons and rodent embryonic stem cell-derived neuronal networks were recorded from the differentiated cultures until up to 4 months. The dependence of the observed neuronal network activity on sodium ion channels was examined using tetrodotoxin (TTX). Antagonists for the glutamate receptors NMDA [D(-)-2-amino-5-phosphonopentanoic acid] and AMPA/kainate [6-cyano-7-nitroquinoxaline-2,3-dione], and for GABAA receptors [(-)-bicuculline methiodide] modulated the spontaneous electrical activity, indicating that pharmacologically susceptible neuronal networks with functional synapses had been generated. The findings indicate that hESC-derived neuronal cells can generate spontaneously active networks with synchronous communication in vitro, and are therefore suitable for use in developmental and drug screening studies, as well as for regenerative medicine.

  7. Stromal cell-derived factor-1α promotes angiogenesis in the peri-infarct region in adults with cerebral infarction

    Institute of Scientific and Technical Information of China (English)

    凌莉

    2014-01-01

    Objective To investigate the possible effects of exogenous stromal cell-derived factor-1α(SDF-1α)on cell proliferation and angiogenesis in the ipsilateral thalamic ventroposterior nucleus(VPN)in adult rats with focal cortical infarction.Methods Thirty-six hypertensive rats with focal cortical infarction were divided randomly into the SDF-1αgroup,vehicle

  8. Induced pluripotent stem cell-derived neuron as a human model for testing environmentally induced developmental neurotoxicity

    Science.gov (United States)

    Induced pluripotent stem cell-derived neurons as a human model for testing environmentally induced developmental neurotoxicity Ingrid L. Druwe1, Timothy J. Shafer2, Kathleen Wallace2, Pablo Valdivia3 ,and William R. Mundy2. 1University of North Carolina, Curriculum in Toxicology...

  9. Integrated Analysis of Contractile Kinetics, Force Generation, and Electrical Activity in Single Human Stem Cell-Derived Cardiomyocytes

    NARCIS (Netherlands)

    Kijlstra, Jan David; Hu, Dongjian; Mittal, Nikhil; Kausel, Eduardo; van der Meer, Peter; Garakani, Arman; Domian, Ibrahim J.

    2015-01-01

    The quantitative analysis of cardiomyocyte function is essential for stem cell-based approaches for the in vitro study of human cardiac physiology and pathophysiology. We present a method to comprehensively assess the function of single human pluripotent stem cell-derived cardiomyocyte (hPSC-CMs) th

  10. IL-2基因转导CD3AK细胞免疫学功能的研究%Research on Immunologic Functions of Interleukin-2 Gene Transducted CD3AK Cells by Retroviral Vector PLIL-2SN

    Institute of Scientific and Technical Information of China (English)

    王立新; 夏圣; 许靖霞; 蔡仙德

    2000-01-01

    目的:观察白细胞介素-2(IL-2)基因转导后CD3AK细胞免疫学功能的变化。方法:应用逆转录病毒载体将IL2基因转导入CD3AK细胞。检测转导细胞中特异性NeoR基因、培养上清IL2的表达水平及转导CD3AK细胞的体外增殖活性、细胞毒活性和细胞表型。结果:从转导细胞mRNA中扩增出长度为347bp的特异性NeoR基因片段,转导细胞培养上清的IL2表达水平显著增高,体外增殖活性和细胞毒活性均强于未转导组细胞,CD4+/CD2+值升高。结论:PLIL2SN逆转录病毒转导CD3AK细胞后,IL-2基因得到表达并增强CD3AK细胞的免疫学功能。%Objective This experiment was designed to observe the immunologic functions of CD3AK cells into which interleukin-2(IL-2) gene had been transducted. Methods The post-transfer CD3AK cells' special NeoR gene and cell immunologic functions including IL-2 expression, proliferation, cytotoxicity and cell phonetype were detected. R~ults The specific 347 bp NeoR gene was amplfiied in post-transfer cells. The post-transfer cells expressed higher IL-2, proliferation and cytotoxicity ability. It was also found that the ratio of CD4 + T cell to CDa + T cell increased in post-transfer group. Conclusion Transducting IL-2 gene into CD3AK cells could enhance their immunologic functions.

  11. Study on Gene Cloning and Prokaryotic Expression of Human Interleukin-2%人源白细胞介素(h-IL2)基因克隆及原核表达的研究

    Institute of Scientific and Technical Information of China (English)

    刘钦松; 张丛丛; 刘孟刚; 许彤

    2011-01-01

    To construct a recombinant espression vector of human Interleukin-2, and eapress in prokaryotic cells, which is helpful for the further purification of recombinant proteins and antibodies production. Methods; Am plify the h-JL2 gene by PCR,the DNA fragments were cloned into pMD-18T simple vectors,transformed into compe tent DH5a,recombinant plasmid PCR,restriction enzyme digest and sequencing confirmed that the fragment was h IL2 and the sequence was identical to that published in GenBank. (NM_000586. 3). Then the h-IL2 gene were cloned into pGEX-4T-l constructing expression vectors, transformed into competent BL21(DE3) , induced by IPTG, SDS-PAGE verification successful expression of the fusion protein GST-IL2.%构建人源白介素(h-IL2)基因的表达载体,并尝试在原核细胞中诱导表达,以用于进一步重组蛋白纯化和抗体生产.以h-IL2基因为模板,采用PCR技术扩增h-IL2基因,连接到pMD-18T载体中构建克隆载体,转化到感受态DH5α,提取质粒PCR、双酶切验证连接成功,经DNA测序鉴定基因序列与GenBank数据库收录h-IL2基因(NM_000586.3)序列一致;再将h-IL2基因插入到pGEX-4T-1构建表达载体,转化至感受态BL21( DE3),筛选阳性克隆IPTG诱导.SDS-PAGE验证成功表达融合蛋白GST-IL2.

  12. Effect of stromal-cell-derived factor 1 on stem-cell homing and tissue regeneration in ischaemic cardiomyopathy

    Science.gov (United States)

    Askari, Arman T.; Unzek, Samuel; Popovic, Zoran B.; Goldman, Corey K.; Forudi, Farhad; Kiedrowski, Matthew; Rovner, Aleksandr; Ellis, Stephen G.; Thomas, James D.; DiCorleto, Paul E.; Topol, Eric J.; Penn, Marc S.

    2003-01-01

    BACKGROUND: Myocardial regeneration via stem-cell mobilisation at the time of myocardial infarction is known to occur, although the mechanism for stem-cell homing to infarcted tissue subsequently and whether this approach can be used for treatment of ischaemic cardiomyopathy are unknown. We investigated these issues in a Lewis rat model (ligation of the left anterior descending artery) of ischaemic cardiomyopathy. METHODS: We studied the effects of stem-cell mobilisation by use of granulocyte colony-stimulating factor (filgrastim) with or without transplantation of syngeneic cells. Shortening fraction and myocardial strain by tissue doppler imaging were quantified by echocardiography. FINDINGS: Stem-cell mobilisation with filgrastim alone did not lead to engraftment of bone-marrow-derived cells. Stromal-cell-derived factor 1 (SDF-1), required for stem-cell homing to bone marrow, was upregulated immediately after myocardial infarction and downregulated within 7 days. 8 weeks after myocardial infarction, transplantation into the peri-infarct zone of syngeneic cardiac fibroblasts stably transfected to express SDF-1 induced homing of CD117-positive stem cells to injured myocardium after filgrastim administration (control vs SDF-1-expressing cardiac fibroblasts mean 7.2 [SD 3.4] vs 33.2 [6.0] cells/mm2, n=4 per group, p<0.02) resulting in greater left-ventricular mass (1.24 [0.29] vs 1.57 [0.27] g) and better cardiac function (shortening fraction 9.2 [4.9] vs 17.2 [4.2]%, n=8 per group, p<0.05). INTERPRETATION: These findings show that SDF-1 is sufficient to induce therapeutic stem-cell homing to injured myocardium and suggest a strategy for directed stem-cell engraftment into injured tissues. Our findings also indicate that therapeutic strategies focused on stem-cell mobilisation for regeneration of myocardial tissue must be initiated within days of myocardial infarction unless signalling for stem-cell homing is re-established.

  13. IL-2 and IFN-gamma, but not IL-4 secretion by peripheral blood mononuclear cells (PBMC) are related to CD4+ T cells and clinical status in Brazilian HIV-1-infected subjects.

    Science.gov (United States)

    Hong, M A; Wakim, V L; Salomão, S J; Camargo, L S; Casseb, J; Duarte, A J

    1998-01-01

    It has been reported that production of IL-2 and IFN-gamma, known as T-helper type 1 cytokines, by peripheral mononuclear cells (PBMC) decreases with progression of HIV infection. In contrast, IL-4 and IL-10 production, Th2 cytokine profile, increases with HIV disease progression. PBMC were evaluated from 55 HIV-infected subjects from Divisão de Imunologia, Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, to "in vitro" cytokines production after 24 hours of stimulation with PHA. Low levels of IL-4 production in both HIV-infected patients and normal subjects, were detected. The patients with CD4+ T cell counts 500 cells/mm3) also showed decreased production of IL-2 that was not statistically significant. There was a correlation between IL-2 and IFN-gamma release with CD4+ T cells counts. HIV-1-infected individuals with CD4+ T cells > 500 cells/mm3 showed increased levels of IL-2 and IFN-gamma, than individuals with CD4+ T cells < 500 cells/mm3. In conclusion, we observed a decline of IL-2 and IFN-gamma production at advanced HIV disease. IL-4 production was not affected during HIV infection. Taken together, these findings suggest that the cytokine profile might be influenced by the HIV infection rather than the cause of disease progression.

  14. CD54/intercellular adhesion molecule 1 and major histocompatibility complex II signaling induces B cells to express interleukin 2 receptors and complements help provided through CD40 ligation

    DEFF Research Database (Denmark)

    Poudrier, J; Owens, T

    1994-01-01

    We have examined signaling roles for CD54 intercellular adhesion molecule 1 and major histocompatibility complex (MHC) II as contact ligands during T help for B cell activation. We used a T helper 1 (Th1)-dependent helper system that was previously shown to be contact as well as interleukin 2 (IL-2......) dependent to demonstrate the relative roles of CD54, MHC II, and CD40 signaling in the events leading to the induction of B cell proliferation and responsiveness to IL-2. Paraformaldehyde-fixed activated Th1-induced expression of IL-2R alpha, IL-2R beta, and B7, and upregulated MHC II and CD54 on B cells...

  15. Effect of Moxibustion of Guan Yuan (RN-4)and Zu San Li (ST-36) on the Cytokine IL-2 in Rats with Epilepsy%艾灸关元和足三里穴对癫痫大鼠细胞因子IL-2影响的研究

    Institute of Scientific and Technical Information of China (English)

    张余山; 张艳晓; 黄静波; 程为平

    2013-01-01

    Objective:To study the effect of moxibustion of Guan Yuan (RN -4) and Zu San Li (ST-36) on changes in serum IL - 2 levels through performing moxibustion on RN - 4 and ST - 36 on rats with pentylenetet-razol induced epilepsy. Methods: Healthy adult female SD rats were injected intraperitoneally with pentylenetet-razol kindling 35.0mg/kg to induce a rat epilepsy model. Behaviour was then analyzed and recorded by using the Racine six point grading scale. After successful construction of the epilepsy model, the rats were divided into four groups, a blank group (three rats) , model group (eight rats) , medication group ( eight rats) , and moxibustion group ( eight rats). The rats in the moxibustion group had moxibustion performed on RN - 4 and bilateral ST-36 for 15 minutes, and the rats in the medication group were given 15.75mg/kg of sodium valproate by gavage once a day continuously for 20 days. The model group and blank group did not receive any form of treatment. After treatment, enzyme linked immunoassay was used to measure serum IL -2 levels. Results:There was a significant difference between the blank group and model group, and between the model group and moxibustion group(P 0. 05). Conclusion: Moxibustion on RN -4 and ST -36 is effective for treating epilepsy in rats. The mechanism may have a correlation with a rise in IL - 2 levels.%目的:通过艾灸戊四氮癫痫大鼠关元、足三里穴,研究艾灸关元、足三里穴对于癫痫大鼠血清IL-2含量变化的影响.方法:取健康成年雌性SD大鼠腹腔注射戊四氮35.0 mg/kg诱发大鼠点燃致痫模型,参照Racine的6级评分标准,记录行为学表现.造模成功后分为空白组(3只)、模型组(8只)、药物组(8只)和艾灸组(8只)4组,对艾灸组大鼠艾灸关元及双侧足三里穴15 min,药物组给予15.75mg/kg丙戊酸钠灌胃治疗.每日1次,治疗20天.模型组、空白组不给予任何治疗.治疗结束后,用酶联免疫法测量血清中IL-2含量.结果:空

  16. Effect of Pingmin Granule (PMG) on Chronic Eczema and Its Impact on Serum Levels of TNF-2α, sIL-2R and ECP%平敏冲剂治疗慢性湿疹及其对患者血清中TNF-2α,sIL-2R和ECP水平的影响

    Institute of Scientific and Technical Information of China (English)

    曾金贵; 喻国华; 刘建国

    2012-01-01

    目的:观察平敏冲剂对慢性湿疹患者的疗效及其对血清肿瘤坏死因子2α(TNF-2α)、可溶性白细胞介素2受体(sIL-2R)、嗜酸细胞阳离子蛋白(ECP)水平的影响.方法:将71例慢性湿疹患者随机分为平敏冲剂组与西替利嗪组,采用ELISA法检测两组治疗前后血清TNF-2α,sIL-2R及ECP水平,并观察疗效.结果:平敏冲剂组有效率86.1%,西替利嗪组有效率54.3%,两组比较差异有统计学意义(P<0.05);且平敏冲剂组复发率8.0%低于西替利嗪组41.4%,两组比较差异有统计学意义(P<0.05).慢性湿疹患者血清TNF-2α,sIL-2R及ECP水平较健康组明显升高(P<0.01),平敏冲剂组与西替利嗪组比较能显著降低患者血清中的TNF-2α[(36.93±9.51),(42.83±11.72)ng·L-1,(P<0.05)],sIL-2R[(46.42±10.61),(53.34±15.66) μg·L-1,(P<0.05)],ECP[(25.81±9.64),(30.16 ±8.68)μg·L-1,(P <0.05)].结论:平敏冲剂对慢性湿疹具有良好的疗效,调节TNF-2α,sIL-2R与ECP的血清水平可能是其作用机制.%Objective: To study effect of Pingmin Granule (PMG) on chronic eczema and its impact on tumor necrosis factor 2α (TNF-2α) , soluble inter leukin 2 thereceptor (slL-2R) and eosinophil cationic protein ( ECP) levels. Method: The effects of PMG and cetirizine on chronic eczema patients in two groups were observed respectively, and the serum levels of TNF-2α, slL-2R and ECP were measured by ELISA before and after treatment. Result: In PMG group effective rate was 86. 1% vs 54. 3% (cetirizine) and recurrence rate was 8. 0% vs 44. 4% ( cetirizine) ( all P < 0. 05 ) . Serum levels of TNF-2α, slL-2R and ECP in patients were higher than those in normal subjects ( P < 0. 01 ) , and PMG group could be significantly lower than cetirizine group in the serum of.patients with TNF-2α [ (36.93 ±9.51), (42.83 ± 11.72) ng ·L-1, (P < 0. 05) ], sIL-2R [ (46.42 ±10.61), (53.34±15.66) μg·L-1, (P<0.05)], ECP [ (25.81±9.64) , (30.16±8.68) μg · L-1, (P <0. 05

  17. El factor de transferencia como inductor de la expresión de RNAm de IFN-γ e IL-2 en pollos vacunados contra influenza aviar Transfer factor acting as IFN-γ and IL-2 mRNA expression inductor in chicken vaccinated against avian influenza

    Directory of Open Access Journals (Sweden)

    A Bravo-Blas

    2010-01-01

    Full Text Available La influenza aviar es una enfermedad de gran importancia económica para la industria avícola. En México sólo se ha reportado la cepa H5N2 de baja patogenicidad y ésta se controla mediante la vacunación con virus inactivado. Esta vacuna en emulsión reduce la presencia de signos, pero no la eliminación viral. Desde hace más de 50 años se ha informado acerca de la eficacia del Factor de Transferencia (FT como inmunomodulador en casos clínicos humanos y en menor cantidad en modelos animales. El objetivo de este trabajo fue el de establecer la dosis que produce un mayor porcentaje de expresión del RNAm de dos citocinas: IL-2 y de IFN-γ. Se diseñó un experimento para evidenciar la expresión del RNAm de estas dos citocinas en pollos previamente inoculados con FT específico para influenza aviar. En la primera fase se aplicaron 0,1, 1, y 10 unidades de FT a diferentes grupos de pollos, posteriormente se realizó la PCR a partir de tejido esplénico. En la segunda fase se aplicó el FT junto con la vacuna a tres nuevos grupos de pollos. Del experimento 1 solamente IL-2 tuvo un porcentaje mayor de positivos (58,33% con 1 unidad (P Avian influenza is a disease of paramount economical importance for the poultry industry. In Mexico, only low pathogenicity H5N2 strain has been reported and it is controlled through inactivated-virus inoculation. This emulsified vaccine reduces clinical signs indeed, but not viral shedding. Over the last 50 years Transfer Factor (TF has shown to be an efficient immunomodulator and has been used successfully in human clinical cases, and less commonly in animal models. The aim of this work was to establish an avian influenza-specific TF dose able to produce the highest percentage of mRNA expression of the following cytokines: IL-2 and IFN-γ. An experiment to show the mRNA expression of these cytokines in chicken previously inoculated with avian influenza-specific TF was set up. In the first experiment 0.1, 1 and

  18. Otimização de metodologia PCR-SSP para identificação de polimorfismos genéticos de TNF e IL2 Optimization of the PCR-SSP methodology in the identification of TNF and IL2 genetic polymorphisms

    Directory of Open Access Journals (Sweden)

    Danilo A. S. Franceschi

    2009-08-01

    Full Text Available A análise de polimorfismos únicos de nucleotídeos (SNPs de citocinas pode ser útil em estudos de frequências alélicas e genotípicas em populações saudáveis de diversas regiões, em estudos de associação com doenças infecciosas ou autoimunes, em estudos antropológicos e na evolução pós-transplante. Estes SNPs podem ser avaliados por diferentes métodos moleculares. O objetivo deste estudo foi aperfeiçoar uma metodologia PCR-SSP simples e rápida para a genotipagem de três SNPs de citocinas usando um único teste laboratorial. Para a identificação de IL2-330T/G e IL2+166G/T foram utilizados dois procedimentos na mesma genotipagem, cada um baseado no uso de quatro iniciadores. Para a detecção de TNF-238G/A foram utilizados dois iniciadores que amplificam a guanina e adenina na posição -238. Este estudo permitiu aperfeiçoar um método simples e rápido para identificar três SNPs de citocinas num único teste, podendo ser utilizado em qualquer laboratório de biologia molecular, como alternativa ao uso de kits de alto custo.The analysis of cytokine single nucleotide polymorphisms (SNPs can be useful in studies of allelic and genotypic frequencies in healthy populations from different regions of Brazil, in association studies of infectious or auto-immune diseases, in anthropological studies and in studies on post-transplant evolution. These SNPs can be assessed by different molecular methods. The objective of this study was to improve a simple and fast methodology, PCR-SSP, for the genotyping of three cytokine SNPs using a single laboratorial test. To identify IL2-330T/G and IL2+166G/T, two procedures were used in the same genotyping assay, each one based on the use of 4 primers. To detect TNF-238G/A, two primers were used that amplify guanine and adenine at position -238. This study enabled the improvement of a simple and fast method for identifying three cytokine SNPs in a single test, which can be adopted in any Molecular

  19. 高乌甲素对脑外伤大鼠IL-1、IL-2、TNF-α水平的影响%Changes of IL-1, IL-2 and TNF-α in response to lappaconitine in rats with traumatic brain injury

    Institute of Scientific and Technical Information of China (English)

    欧珊; 林露; 李军; 周乐顺; 匡永勤; 顾建文; 王红梅; 安虹

    2012-01-01

    目的 观察高乌甲素( lappaconitine,LA)对创伤性脑损伤(traumatic brain injury,TBI)大鼠脑含水量以及血清IL-1、IL -2和TNF -a水平的影响及其神经保护功能. 方法 24只雄性SD大鼠按随机数字表法分为对照组、TBI组和TBI+ LA组,每组8只.TBI组和TBI+ LA 组制作大鼠颅脑液压打击伤(FPI)模型,TBI+ LA组大鼠接受腹腔注射LA(4 mg/kg,连续10 d)处理,在T0(FPI前)、TI(FPI后1d)、T2(FPI后5d)和T3(FPI后10d)时相点对各组大鼠进行神经功能评分、脑含水量测定和血清IL-1、IL -2、TNF -α浓度检测. 结果 FPI后各时相点,TBI组和TBI+ LA组大鼠均出现明显的神经功能障碍,但从TI到T3,TBI+ LA组神经功能障碍逐渐减轻,神经功能评分明显低于TBI组(P<0.05或0.01).FPI后各时相点,TBI组和TBI+ LA组大鼠脑含水量明显高于对照组,同时TBI +LA组含水量明显低于TBI组(P<0.01);TBI组和TBI+ LA 组血清IL-1、IL -2、TNF -α浓度明显高于对照组,同时TBI+ LA组各血清细胞因子水平均明显低于TBI组(P<0.01). 结论 LA可减轻TBI大鼠的神经功能障碍和脑水肿程度,降低血清IL -1、IL -2、TNF -α浓度,从而发挥对TBI大鼠的脑保护作用.%Objective To observe the effect of lappaconitine (LA) on brain water content and serum IL-I,IL-2 and TNF-α levels in rats with traumatic brain injury (TBI) as well as its cerebral protective function. Methods A total of 24 male SD rats were involved in the study and randomly divided into control group,TBI group and TBI +LA group,with eight rats per group.The rats in the TBI group and TBI + LA group were inflicted with fluid percussion injury ( FPI ).The rats in the TBI + LA group were treated with LA (4 mg/Kg/d,ip,for 10 consecutive days).The neurological score,brain water content and serum IL-I,IL-2 and TNF -α concentrations were detected at time points including TO ( before FPI ),T1 (one day after FPI),T2 (five days after FPI) andT3 (10 days after FPI). Results At

  20. 佛波醇酯加离子霉素诱导脐血和成人外周血CD4+CD25+T细胞分泌IL-2的相关机制研究%Mechanisms underlying the induction of IL- 2 secretion by PDB plus ionomycin in CD4 + CD25 + T cells from cord blood and adult peripheral blood

    Institute of Scientific and Technical Information of China (English)

    肇静娴; 曾耀英; 李海仙; 曾祥凤; 季煜华; 何贤辉

    2006-01-01

    目的:以佛波醇酯加离子霉素作为刺激剂,验证CD4+CD25+调节性T细胞本身并不存在分泌IL-2障碍;同时通过对脐血和成人外周血的比较性研究,了解脐血CD4+CD25+T细胞的成熟度.方法:以autoMACS从足月婴儿脐血(CB)和成人外周血(PB)分选CD4+CD25+和CD4+CD25-T细胞,以PDB+ionomycin作为刺激剂,培养45h后流式细胞术检测各组细胞表达CD69和CD25水平,并以Luminex多重细胞因子检测技术检测培养上清中7种细胞因子的浓度.结果:经PDB+ionomycin刺激后,CB、PB的CD4+CD25+和CD4+CD25-T细胞均发生增殖,但在培养45h后CD4+CD25+T细胞均出现细胞状态变差或死亡倾向.CB、PB的CD4+CD25+T细胞活化后CD25分子表达进一步上调,高于CD25-细胞活化后的CD25分子密度.经PDB+ionomycin刺激后,PB CD4+CD25+和CD4+CD25-T细胞均分泌高水平的IFN-γ、IL-2和TNF-α,但CD25+细胞分泌IL-5、IL-4和IL-10水平远远高于CD25-细胞;CBCD4+CD25+和CD4+CD25-T细胞亦分泌高水平的IL-2和TNF-α,但IFN-γ水平远远低于PB,基本不分泌IL-5、IL-4和IL-10.结论:CD4+CD25+T细胞本身并不存在合成和分泌IL-2障碍,其可能具有与传统T细胞不同的T细胞受体信息转导模式;脐血CD4+CD25+T细胞功能尚未完全成熟.

  1. Mononucleosis and Antigen-Driven T Cell Responses Have Different Requirements for Interleukin-2 Signaling in Murine Gammaherpesvirus Infection ▿ †

    OpenAIRE

    Molloy, Michael; Zhang, Weijun; Usherwood, Edward

    2010-01-01

    Interleukin-2 (IL-2) has been implicated as being necessary for the optimal formation of primary CD8+ T cell responses against various pathogens. Here we have examined the role that IL-2 signaling plays in several aspects of a CD8+ T cell response against murine gammaherpesvirus 68 (MHV-68). Exposure to MHV-68 causes a persistent infection, along with infectious mononucleosis, providing a model for studying these processes in mice. Our study indicates that CD25 is necessary for optimal expans...

  2. Exposure to phthalates affects calcium handling and intercellular connectivity of human stem cell-derived cardiomyocytes.

    Directory of Open Access Journals (Sweden)

    Nikki Gillum Posnack

    Full Text Available The pervasive nature of plastics has raised concerns about the impact of continuous exposure to plastic additives on human health. Of particular concern is the use of phthalates in the production of flexible polyvinyl chloride (PVC products. Di-2-ethylhexyl-phthalate (DEHP is a commonly used phthalate ester plasticizer that imparts flexibility and elasticity to PVC products. Recent epidemiological studies have reported correlations between urinary phthalate concentrations and cardiovascular disease, including an increased risk of high blood pressure and coronary risk. Yet, there is little direct evidence linking phthalate exposure to adverse effects in human cells, including cardiomyocytes.The effect of DEHP on calcium handling was examined using monolayers of gCAMP3 human embryonic stem cell-derived cardiomyocytes, which contain an endogenous calcium sensor. Cardiomyocytes were exposed to DEHP (5 - 50 μg/mL, and calcium transients were recorded using a Zeiss confocal imaging system. DEHP exposure (24 - 72 hr had a negative chronotropic and inotropic effect on cardiomyocytes, increased the minimum threshold voltage required for external pacing, and modified connexin-43 expression. Application of Wy-14,643 (100 μM, an agonist for the peroxisome proliferator-activated receptor alpha, did not replicate DEHP's effects on calcium transient morphology or spontaneous beating rate.Phthalates can affect the normal physiology of human cardiomyocytes, including DEHP elicited perturbations in cardiac calcium handling and intercellular connectivity. Our findings call for additional studies to clarify the extent by which phthalate exposure can alter cardiac function, particularly in vulnerable patient populations who are at risk for high phthalate exposure.

  3. Pharmacoelectrophysiology of viral-free induced pluripotent stem cell-derived human cardiomyocytes.

    Science.gov (United States)

    Mehta, Ashish; Chung, YingYing; Sequiera, Glen Lester; Wong, Philip; Liew, Reginald; Shim, Winston

    2013-02-01

    Development of pharmaceutical agents for cardiac indication demands elaborate safety screening in which assessing repolarization of cardiac cells remains a critical path in risk evaluations. An efficient platform for evaluating cardiac repolarization in vitro significantly facilitates drug developmental programs. In a proof of principle study, we examined the effect of antiarrhythmogenic drugs (Vaughan Williams class I-IV) and noncardiac active drugs (terfenadine and cisapride) on the repolarization profile of viral-free human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Extracellular field potential (FP) recording using microelectrode arrays demonstrated significant delayed repolarization as prolonged corrected FP durations (cFPDs) by class I (quinidine and flecainide), class III (sotalol and amiodarone), and class IV (verapamil), whereas class II drugs (propranolol and nadolol) had no effects. Consistent with their sodium channel-blocking ability, class I drugs also significantly reduced FPmin and conduction velocity. Although lidocaine (class IB) had no effects on cFPDs, verapamil shortened cFPD and FPmin by 25 and 50%, respectively. Furthermore, verapamil reduced beating frequencies drastically. Importantly, the examined drugs exhibited dose-response curve on prolongation of cFPDs at an effective range that correlated significantly with therapeutic plasma concentrations achieved clinically. Consistent with clinical outcomes, drug-induced arrhythmia of tachycardia and bigeminy-like waveforms by quinidine, flecainide, and sotalol was demonstrated at supraphysiological concentrations. Furthermore, off-target effects of terfenadine and cisapride on cFPD and Na( + ) channel blockage were similarly revealed. These results suggest that hiPSC-CMs may be useful for safety evaluation of cardioactive and noncardiac acting drugs for personalized medicine.

  4. Radiation response of mesenchymal stem cells derived from bone marrow and human pluripotent stem cells

    International Nuclear Information System (INIS)

    Mesenchymal stem cells (MSCs) isolated from human pluripotent stem cells are comparable with bone marrow-derived MSCs in their function and immunophenotype. The purpose of this exploratory study was comparative evaluation of the radiation responses of mesenchymal stem cells derived from bone marrow- (BMMSCs) and from human embryonic stem cells (hESMSCs). BMMSCs and hESMSCs were irradiated at 0 Gy (control) to 16 Gy using a linear accelerator commonly used for cancer treatment. Cells were harvested immediately after irradiation, and at 1 and 5 days after irradiation. Cell cycle analysis, colony forming ability (CFU-F), differentiation ability, and expression of osteogenic-specific runt-related transcription factor 2 (RUNX2), adipogenic peroxisome proliferator-activated receptor gamma (PPARγ), oxidative stress-specific dismutase-1 (SOD1) and Glutathione peroxidase (GPX1) were analyzed. Irradiation arrested cell cycle progression in BMMSCs and hESMSCs. Colony formation ability of irradiated MSCs decreased in a dose-dependent manner. Irradiated hESMSCs showed higher adipogenic differentiation compared with BMMSCs, together with an increase in the adipogenic PPARγ expression. PPARγ expression was upregulated as early as 4 h after irradiation, along with the expression of SOD1. More than 70% downregulation was found in Wnt3A, Wnt4, Wnt7A, Wnt10A and Wnt11 in BMMSCs, but not in hESMSCs. hESMSCs are highly proliferative but radiosensitive compared with BMMSCs. Increased PPARγ expression relative to RUNX2 and downregulation of Wnt ligands in irradiated MSCs suggest Wnt mediated the fate determination of irradiated MSCs. (author)

  5. Modeling chemotherapeutic neurotoxicity with human induced pluripotent stem cell-derived neuronal cells.

    Directory of Open Access Journals (Sweden)

    Heather E Wheeler

    Full Text Available There are no effective agents to prevent or treat chemotherapy-induced peripheral neuropathy (CIPN, the most common non-hematologic toxicity of chemotherapy. Therefore, we sought to evaluate the utility of human neuron-like cells derived from induced pluripotent stem cells (iPSCs as a means to study CIPN. We used high content imaging measurements of neurite outgrowth phenotypes to compare the changes that occur to iPSC-derived neuronal cells among drugs and among individuals in response to several classes of chemotherapeutics. Upon treatment of these neuronal cells with the neurotoxic drug paclitaxel, vincristine or cisplatin, we identified significant differences in five morphological phenotypes among drugs, including total outgrowth, mean/median/maximum process length, and mean outgrowth intensity (P < 0.05. The differences in damage among drugs reflect differences in their mechanisms of action and clinical CIPN manifestations. We show the potential of the model for gene perturbation studies by demonstrating decreased expression of TUBB2A results in significantly increased sensitivity of neurons to paclitaxel (0.23 ± 0.06 decrease in total neurite outgrowth, P = 0.011. The variance in several neurite outgrowth and apoptotic phenotypes upon treatment with one of the neurotoxic drugs is significantly greater between than within neurons derived from four different individuals (P < 0.05, demonstrating the potential of iPSC-derived neurons as a genetically diverse model for CIPN. The human neuron model will allow both for mechanistic studies of specific genes and genetic variants discovered in clinical studies and for screening of new drugs to prevent or treat CIPN.

  6. Targeting eradication of malignant cells derived from human bone marrow mesenchymal stromal cells

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Yingbin [Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Chongqing 400044 (China); School of Life Science, Southwest University, Chongqing 400715 (China); Cai, Shaoxi, E-mail: sxcai@cqu.edu.cn [Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Chongqing 400044 (China); Yang, Li [Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Chongqing 400044 (China); College of Pharmacy, Jinan University, Guangzhou 510632 (China); Yu, Shuhui [Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Chongqing 400044 (China); Library of Southwest University, Chongqing 400715 (China); Jiang, Jiahuan; Yan, Xiaoqing [Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Chongqing 400044 (China); Zhang, Haoxing [School of Life Science, Southwest University, Chongqing 400715 (China); Liu, Lan [Department of Laboratory of Medicine, Children' s Hospital of Chongqin Medical University, Chongqing 400014 (China); Liu, Qun [College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041 (China); Du, Jun [Center of Microbiology, Biochemistry, and Pharmacology, School of Pharmaceutical Science, Sun Yat-Sen University, Guangzhou 510080 (China); Cai, Shaohui [College of Pharmacy, Jinan University, Guangzhou 510632 (China); Sung, K.L. Paul [Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Chongqing 400044 (China); Departments of Orthopaedic Surgery and Bioengineering, University of California, SD 0412 (United States)

    2010-12-10

    Human bone marrow mesenchymal stromal cells (hBMSC) have been shown to participate in malignant transformation. However, hampered by the low frequency of malignant transformation of hBMSC, we do not yet know how to prevent malignant transformation of implanted hBMSC. In this study, in order to establish a model for the eradication of hBMSC-derived malignant cells, a gene fusion consisting of a human telomerase (hTERT) promoter modified with both c-Myc and myeloid zinc finger protein2 (MZF-2) binding elements and followed by the E. coli cytosine deaminase (CD) and luciferase genes was stably transferred into hBMSC via lentiviral transduction; n-phosphonacelyl-L-aspartic acid (PALA) selection was used to generate malignant cell colonies derived from transduced hBMSC after treatment with the carcinogenic reagent BPDE. Cells that were amplified after PALA selection were used for transplantation and 5-FC pro-drug cytotoxicity tests. The results showed that PALA-resistant malignant cells could be generated from hBMSC co-induced with lentiviral transduction and treatment with Benzo(a)pyrene Diol Epoxide (BPDE); the modification of c-Myc and MZF-2 binding elements could remarkably enhance the transcriptional activities of the hTERT promoter in malignant cells, whereas transcriptional activity was depressed in normal hBMSC; malignant cells stably expressing CD under the control of the modified hTERT promoter could be eliminated by 5-FC administration. This study has provided a method for targeted eradication of malignant cells derived from hBMSC.

  7. Targeting eradication of malignant cells derived from human bone marrow mesenchymal stromal cells

    International Nuclear Information System (INIS)

    Human bone marrow mesenchymal stromal cells (hBMSC) have been shown to participate in malignant transformation. However, hampered by the low frequency of malignant transformation of hBMSC, we do not yet know how to prevent malignant transformation of implanted hBMSC. In this study, in order to establish a model for the eradication of hBMSC-derived malignant cells, a gene fusion consisting of a human telomerase (hTERT) promoter modified with both c-Myc and myeloid zinc finger protein2 (MZF-2) binding elements and followed by the E. coli cytosine deaminase (CD) and luciferase genes was stably transferred into hBMSC via lentiviral transduction; n-phosphonacelyl-L-aspartic acid (PALA) selection was used to generate malignant cell colonies derived from transduced hBMSC after treatment with the carcinogenic reagent BPDE. Cells that were amplified after PALA selection were used for transplantation and 5-FC pro-drug cytotoxicity tests. The results showed that PALA-resistant malignant cells could be generated from hBMSC co-induced with lent