- Email: [email protected]
ID: 21
Abstract / Cytokine 76 (2015) 66–112 IL-27Ra and gp130, IL-27 did not inhibit cell growth, but caused a dose-dependent proliferation of the acute myeloid leukemic cell line OCI-AML5, and the erythroleukemic cell lines, TF-1, UT-7 and UT7/EPO. Consistent with this, IL-27 promoted cell survival and reduced TNF-a-induced apoptosis of the leukemic cell lines. IL-27 also decreased the sensitivity of the leukemic cells to chemotherapeutic drugs cytarabine and daunorubicin. We observed that IL-27 induced the activation of STAT1/3 and ERK1/2 in the leukemic cells. Growth stimulation by IL-27 was suppressed by the specific MEK inhibitor U0126 and the PI3K inhibitor wortmannin, suggesting that IL-27-induced cell proliferation is mediated via the activation of two distinct pathways of MAPK/ERK and PI3K/AKT. The present study is the first demonstration of the proliferative property of IL-27 in human leukemic cell lines, suggesting that IL27 may play an unfavourable role in tumour growth of certain subtypes of human leukaemia.
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observed, at this cell division stage, an increased activity and phosphorylation of TBK1 that lead to its relocalization to mitochondria and to enhanced interferon production, and activation of the antiviral innate immune response before cells enter into mitosis. Overall our study shows that the antiviral immune system is controlled during the cell cycle and that Optineurin-mediated induction of this system might serve to protect cells from infections that eventually occur during cell division. http://dx.doi.org/10.1016/j.cyto.2015.08.050
ID: 21 Ganoderma lucidum-derived triterpenes inhibit PAR2-mediated IL8 production by human keratinocytes
http://dx.doi.org/10.1016/j.cyto.2015.08.048
ID: 19 Increased heparanase expression in keratinocytes promotes dermal fibrosis in scleroderma Chul-Soo Cho 1,*, In-Woon Baek 1, Ki-Jo Kim 2, Wan-Uk Kim 3, 1 Yeouido St. Mary’s Hospital, The Catholic University of Korea, Seoul, Republic of Korea, 2 St. Vincent’s Hospital, The Catholic University of Korea, Suwon, Republic of Korea, 3 Seoul St. Mary’s Hospital, The Catholic University of Korea, Seoul, Republic of Korea * Corresponding author at: Ludwig Boltzmann Institute for Cancer Research, Austria. Interactions between keratinocyte and dermal fibroblast via paracrine loop play an important role in wound repair and keloid formation. In this study, we investigated heparanase expression in activated keratinocyte, and tested its effect on the survival of dermal fibroblasts. Plasma heparanase levels were measured in scleroderma patients, and heparanase expression was evaluated in the skin of bleomycin-induced fibrosis mice and HaCaT keratinocyte (HaCaT). Dermal fibroblasts were cocultured with HaCaT separated by transwell insert under serum starvation, and apoptosis was determined using APOPercentage assay. Plasma heparanase levels were significantly higher in 26 scleroderma patients than in 10 healthy subjects, and positively correlated with plasma TGF-b levels. In bleomycin-induced fibrosis mice, increased heparanase expression was observed in keratinocyte layer, but not in dermal layer. Treatment of HaCaT with hypoxia resulted in significant increase in heparanase expression, and this increase was accompanied by concomitant increase of matrix metalloproteinase-9, both of which are known to degrade epidermal basement membrane components. Coculture of dermal fibroblasts and HaCaT in the presence of hypoxia significantly protected the apoptosis of dermal fibroblasts induced by serum starvation, but it was abolished by anti-heparanase antibody or transfection of HaCaT with heparanase siRNA. Dermal fibroblasts cocultured with HaCaT exposed to hypoxia exhibited increased Akt phosphorylation, and pretreatment of dermal fibroblasts with LY294002, an inhibitor of phosphatidylinositol 3-kinase, significantly abolished anti-apoptotic effect of heparanase on dermal fibroblasts. These data indicate that hypoxia, caused possibly by microvascular alteration, increases heparanase production in keratinocytes, which may promote fibrosis in scleroderma by inhibiting the apoptosis of dermal fibroblasts. http://dx.doi.org/10.1016/j.cyto.2015.08.049
ID: 20 Optineurin regulates the interferon response in a cell cycle-dependent manner Pierre Génin 1,*, Frédérique Cuvelier 1, Sandrine Lambin 1, Josina Côrte-Real Filipe 1, Elodie Autrusseau 1, Christine Laurent 2, Emmanuel Laplantine 1, Robert Weil 1, 1 Laboratoire de Signalisation et Pathogenèse, CNRS UMR3691, Institut Pasteur, 75724 Paris Cedex 15, France, 2 Plate-Forme Protéomique, Institut Pasteur, 75724 Paris Cedex 15, France * Corresponding author at: Ludwig Boltzmann Institute for Cancer Research, 1090 Vienna, Austria. Viral invasion into a host is initially recognized by the innate immune system, mainly through activation of the intracellular cytosolic signaling pathway and coordinated activation of interferon regulatory factor 3 (IRF3) and nuclear factor kappa B (NF-jB) transcription factors that promote type I interferon gene induction. The TANK-binding Kinase 1 (TBK1) phosphorylates and activates IRF3. We now show that Optineurin (Optn) dampens the antiviral innate immune response by targeting the deubiquitinating enzyme CYLD to TBK1 in order to inhibit its enzymatic activity. Importantly, we found that this regulatory mechanism is abolished at the G2/M phase as a consequence of the nuclear translocation of CYLD and Optn. As a result, we
Yoshio Ishibashi 1,*, Susumu Kitanaka 2, Yuji Asahara 3, 1 Meiji Pharmaceutical University, Japan, 2 Nihon University, Japan, 3 Nissan Chemical Industries, Ltd., Japan * Corresponding author at: University Health Network, Canada. Crude extracts from Ganoderma lucidum, a medical mushroom, are popularly used for treatment of allergic diseases such as atopic dermatitis (AD). Triterpenes are the major active constituents of Ganoderma lucidum. Stimulation of proteaseactivated receptor-2 (PAR2) on cutaneous keratinocytes induces secretion of various immune and inflammatory cytokines including interleukin 8 (IL8), and thereby plays a role in the development of allergic skin inflammation. Thus, PAR2-mediated signaling has been suggested to be a therapeutic target for the treatment of allergic diseases. This study aimed to evaluate the inhibitory effects of Ganoderma triterpenes on PAR2-mediated IL8 secretion from human keratinocytes. Normal human epidermal keratinocytes (NHEK) were stimulated with PAR2-activating peptide (AP) in the presence or absence of Ganoderma triterpenes in vitro. IL8 secretion from NHEK cells was measured by enzyme-linked immunosorbent assay (ELISA). Chemical structures of triterpenes were determined based on spectroscopic evidence and mass spectral data. Among of Ganoderma triterpenes, ganolucidate F exhibited most potent inhibitory activity on PAR2-mediated IL8 secretion from keratinocytes (IC50 = 12.5 lM). Ganolucidic acid ca showed strong inhibitory activity with IC50 value of 29.1 lM. In addition, ganoderic acid g and ganoderic acid h showed remarkable inhibitory effects with IC50 values of 64.9 lM and 109.5 lM, respectively. Elucidation of relationships between chemical structure of triterpenes and inhibitory activity implied that substitution of hydroxy group at C-23, instead of a carbonyl group, may be a very important structural feature for inhibitory activity. Ganoderma lucidum-derived ganoderic acids could be a candidate drug for treatment of allergic skin diseases. http://dx.doi.org/10.1016/j.cyto.2015.08.051
ID: 22 Reduction by SUMO of interferon gamma, but not alpha, transcriptional response Ghizlane Maarifi, Mohamed Ali Maroui, Jacques Dutrieux, Laurent Dianoux, Sébastien Nisole, Mounira K. Chelbi-Alix *, UMR-S1124 INSERM, University Paris Descartes, France * Corresponding author at: University of Turku, Finland. Interferons (IFNs) orchestrate immune defense through induction of hundred of genes. Small ubiquitin-like modifier (SUMO) is involved in various cellular functions, but little is known about its role on IFN responses. Prior work identified SUMOylation of STAT1 as an important mode of regulation of IFNc signaling. Here, we investigated the various roles of SUMO in IFN signaling, transcriptional response and IFN-induced biological effects. We first show that SUMO overexpression leads to STAT1 SUMOylation and to a decrease in IFN-induced STAT1 phosphorylation. This process correlates with lower levels of STAT1 binding to GAS in response to IFNc, while the binding of an ISGF3-like complex to ISRE in response to IFNa is not altered. Interestingly, IFNs exert a negative retro-control on their own signaling by enhancing STAT1 SUMOylation. Furthermore, we show that expression of each SUMO paralog selectively inhibits IFNc-induced IP-10 and TAP1 mRNA expression. Importantly, inhibition of IFNc signaling by SUMO is associated with a decrease of IFNc-induced apoptosis and cell growth inhibition as well as a dramatic reduction of IP-10-induced chemotaxis. Conversely, inhibition of SUMOylation results in a higher IFNc-induced STAT1 phosphorylation and biological responses. In conclusion, our work thus allows the inclusion of SUMO to the list of negative regulators of IFN signaling known to date and posits SUMO as a possible contributor to IFNc resistance. http://dx.doi.org/10.1016/j.cyto.2015.08.052
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observed, at this cell division stage, an increased activity and phosphorylation of TBK1 that lead to its relocalization to mitochondria and to enhanced interferon production, and activation of the antiviral innate immune response before cells enter into mitosis. Overall our study shows that the antiviral immune system is controlled during the cell cycle and that Optineurin-mediated induction of this system might serve to protect cells from infections that eventually occur during cell division. http://dx.doi.org/10.1016/j.cyto.2015.08.050
ID: 21 Ganoderma lucidum-derived triterpenes inhibit PAR2-mediated IL8 production by human keratinocytes
http://dx.doi.org/10.1016/j.cyto.2015.08.048
ID: 19 Increased heparanase expression in keratinocytes promotes dermal fibrosis in scleroderma Chul-Soo Cho 1,*, In-Woon Baek 1, Ki-Jo Kim 2, Wan-Uk Kim 3, 1 Yeouido St. Mary’s Hospital, The Catholic University of Korea, Seoul, Republic of Korea, 2 St. Vincent’s Hospital, The Catholic University of Korea, Suwon, Republic of Korea, 3 Seoul St. Mary’s Hospital, The Catholic University of Korea, Seoul, Republic of Korea * Corresponding author at: Ludwig Boltzmann Institute for Cancer Research, Austria. Interactions between keratinocyte and dermal fibroblast via paracrine loop play an important role in wound repair and keloid formation. In this study, we investigated heparanase expression in activated keratinocyte, and tested its effect on the survival of dermal fibroblasts. Plasma heparanase levels were measured in scleroderma patients, and heparanase expression was evaluated in the skin of bleomycin-induced fibrosis mice and HaCaT keratinocyte (HaCaT). Dermal fibroblasts were cocultured with HaCaT separated by transwell insert under serum starvation, and apoptosis was determined using APOPercentage assay. Plasma heparanase levels were significantly higher in 26 scleroderma patients than in 10 healthy subjects, and positively correlated with plasma TGF-b levels. In bleomycin-induced fibrosis mice, increased heparanase expression was observed in keratinocyte layer, but not in dermal layer. Treatment of HaCaT with hypoxia resulted in significant increase in heparanase expression, and this increase was accompanied by concomitant increase of matrix metalloproteinase-9, both of which are known to degrade epidermal basement membrane components. Coculture of dermal fibroblasts and HaCaT in the presence of hypoxia significantly protected the apoptosis of dermal fibroblasts induced by serum starvation, but it was abolished by anti-heparanase antibody or transfection of HaCaT with heparanase siRNA. Dermal fibroblasts cocultured with HaCaT exposed to hypoxia exhibited increased Akt phosphorylation, and pretreatment of dermal fibroblasts with LY294002, an inhibitor of phosphatidylinositol 3-kinase, significantly abolished anti-apoptotic effect of heparanase on dermal fibroblasts. These data indicate that hypoxia, caused possibly by microvascular alteration, increases heparanase production in keratinocytes, which may promote fibrosis in scleroderma by inhibiting the apoptosis of dermal fibroblasts. http://dx.doi.org/10.1016/j.cyto.2015.08.049
ID: 20 Optineurin regulates the interferon response in a cell cycle-dependent manner Pierre Génin 1,*, Frédérique Cuvelier 1, Sandrine Lambin 1, Josina Côrte-Real Filipe 1, Elodie Autrusseau 1, Christine Laurent 2, Emmanuel Laplantine 1, Robert Weil 1, 1 Laboratoire de Signalisation et Pathogenèse, CNRS UMR3691, Institut Pasteur, 75724 Paris Cedex 15, France, 2 Plate-Forme Protéomique, Institut Pasteur, 75724 Paris Cedex 15, France * Corresponding author at: Ludwig Boltzmann Institute for Cancer Research, 1090 Vienna, Austria. Viral invasion into a host is initially recognized by the innate immune system, mainly through activation of the intracellular cytosolic signaling pathway and coordinated activation of interferon regulatory factor 3 (IRF3) and nuclear factor kappa B (NF-jB) transcription factors that promote type I interferon gene induction. The TANK-binding Kinase 1 (TBK1) phosphorylates and activates IRF3. We now show that Optineurin (Optn) dampens the antiviral innate immune response by targeting the deubiquitinating enzyme CYLD to TBK1 in order to inhibit its enzymatic activity. Importantly, we found that this regulatory mechanism is abolished at the G2/M phase as a consequence of the nuclear translocation of CYLD and Optn. As a result, we
Yoshio Ishibashi 1,*, Susumu Kitanaka 2, Yuji Asahara 3, 1 Meiji Pharmaceutical University, Japan, 2 Nihon University, Japan, 3 Nissan Chemical Industries, Ltd., Japan * Corresponding author at: University Health Network, Canada. Crude extracts from Ganoderma lucidum, a medical mushroom, are popularly used for treatment of allergic diseases such as atopic dermatitis (AD). Triterpenes are the major active constituents of Ganoderma lucidum. Stimulation of proteaseactivated receptor-2 (PAR2) on cutaneous keratinocytes induces secretion of various immune and inflammatory cytokines including interleukin 8 (IL8), and thereby plays a role in the development of allergic skin inflammation. Thus, PAR2-mediated signaling has been suggested to be a therapeutic target for the treatment of allergic diseases. This study aimed to evaluate the inhibitory effects of Ganoderma triterpenes on PAR2-mediated IL8 secretion from human keratinocytes. Normal human epidermal keratinocytes (NHEK) were stimulated with PAR2-activating peptide (AP) in the presence or absence of Ganoderma triterpenes in vitro. IL8 secretion from NHEK cells was measured by enzyme-linked immunosorbent assay (ELISA). Chemical structures of triterpenes were determined based on spectroscopic evidence and mass spectral data. Among of Ganoderma triterpenes, ganolucidate F exhibited most potent inhibitory activity on PAR2-mediated IL8 secretion from keratinocytes (IC50 = 12.5 lM). Ganolucidic acid ca showed strong inhibitory activity with IC50 value of 29.1 lM. In addition, ganoderic acid g and ganoderic acid h showed remarkable inhibitory effects with IC50 values of 64.9 lM and 109.5 lM, respectively. Elucidation of relationships between chemical structure of triterpenes and inhibitory activity implied that substitution of hydroxy group at C-23, instead of a carbonyl group, may be a very important structural feature for inhibitory activity. Ganoderma lucidum-derived ganoderic acids could be a candidate drug for treatment of allergic skin diseases. http://dx.doi.org/10.1016/j.cyto.2015.08.051
ID: 22 Reduction by SUMO of interferon gamma, but not alpha, transcriptional response Ghizlane Maarifi, Mohamed Ali Maroui, Jacques Dutrieux, Laurent Dianoux, Sébastien Nisole, Mounira K. Chelbi-Alix *, UMR-S1124 INSERM, University Paris Descartes, France * Corresponding author at: University of Turku, Finland. Interferons (IFNs) orchestrate immune defense through induction of hundred of genes. Small ubiquitin-like modifier (SUMO) is involved in various cellular functions, but little is known about its role on IFN responses. Prior work identified SUMOylation of STAT1 as an important mode of regulation of IFNc signaling. Here, we investigated the various roles of SUMO in IFN signaling, transcriptional response and IFN-induced biological effects. We first show that SUMO overexpression leads to STAT1 SUMOylation and to a decrease in IFN-induced STAT1 phosphorylation. This process correlates with lower levels of STAT1 binding to GAS in response to IFNc, while the binding of an ISGF3-like complex to ISRE in response to IFNa is not altered. Interestingly, IFNs exert a negative retro-control on their own signaling by enhancing STAT1 SUMOylation. Furthermore, we show that expression of each SUMO paralog selectively inhibits IFNc-induced IP-10 and TAP1 mRNA expression. Importantly, inhibition of IFNc signaling by SUMO is associated with a decrease of IFNc-induced apoptosis and cell growth inhibition as well as a dramatic reduction of IP-10-induced chemotaxis. Conversely, inhibition of SUMOylation results in a higher IFNc-induced STAT1 phosphorylation and biological responses. In conclusion, our work thus allows the inclusion of SUMO to the list of negative regulators of IFN signaling known to date and posits SUMO as a possible contributor to IFNc resistance. http://dx.doi.org/10.1016/j.cyto.2015.08.052