- Email: [email protected]
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Abstract / Cytokine 63 (2013) 243–314
cells and the increase in the ratio Teff/Treg in the tumor environment. In conclusion, the favorable pharmacokinetic/pharmacodynamic profile of IL-15 heterodimer allows for lower dose, simple s.c. delivery, and lowers the possibility of toxicity due to cytokine spike. Preclinical cancer studies suggested that IL-15 heterodimer-based immunotherapy favors the development of anti-tumor responses by favoring cytotoxic over regulatory cells. http://dx.doi.org/10.1016/j.cyto.2013.06.023
21 Interferon-mediated potentiation of SMAC mimetic compound cytotoxicity by oncolytic virotherapy Shawn T. Beug a, Vera A. Tang a, Herman C. Cheung a, Fabrice Le Boeuf b, John C. Bell b, Eric C. LaCasse a, Robert G. Korneluk a, a Apoptosis Research Centre, Children’s Hospital of Eastern Ontario Research Institute, Ottawa, Ont., Canada, b Centre for Innovative Cancer Therapeutics, Ottawa Hospital Research Institute, Ottawa, Ont., Canada Smac mimetic compounds (SMCs) are a novel class of small molecule antagonists that target the inhibitor of apoptosis (IAP) proteins to induce cancer cell death via cytokine activation of caspase-8 death pathways. Oncolytic viruses (OVs) selectively target cancer cells for death via multiple mechanisms, including oncolysis and the induction of anti-tumor immunity. We reasoned that oncolytic viruses such as the rhabdovirus VSVD51 will provide a proinflammatory environment that would augment SMC-induced cancer cell death. We observed that SMC and VSVD51 synergy was specific to cancer cells and this combination increased the potency of cancer cell death up to 10,000-fold compared to stand-alone VSVD51 therapy. The combination resulted in delayed tumor growth and lead to durable cures in mouse models of cancer. Furthermore, several non-viral immunostimulatory agents, such as the viral RNA mimic poly (I:C) or the bacterial DNA mimic CpG, elicit similar responses when combined with SMC treatment. The combination leads to bystander cancer cell death via the production of type 1 interferon-dependent production of proinflammatory cytokines, such as TNFa and TRAIL, which sensitizes cancer cells to SMC-mediated caspase-8-dependent apoptosis. We thus demonstrate the novel combination of targeted immunotherapies for the treatment of cancer, which on their own are limited by various efficacy hurdles, yet when combined, the combined treatment results in strong synergy to cause cancer cell death. http://dx.doi.org/10.1016/j.cyto.2013.06.024
22 The role of STAT1 in macrophages/neutrophils in the response to murine cytomegalovirus infection Mario Biaggio a, Caroline Lassnig a,b, Ursula Reichart a,b, Rita Rom a, Astrid Krmpotic c, Stipan Jonjicì c, Birgit Strobl a, Mathias Müller a,b, a Institute of Animal Breeding and Genetics, University of Veterinary Medicine, Vienna, Austria, b Biomodels Austria, University of Veterinary Medicine, Vienna, Austria, c Department of Histology and Embryology, Faculty of Medicine, University of Rijeka, Croatia Signal transducers and activators of transcription (STAT) 1 is a key transcription factor involved in innate and adaptive immune responses by governing the responses to interferons (IFNs) type I, II and III. In order to elucidate the cell type specific contributions of STAT1 to the host response to viral infections our lab generated conditional Stat1 mice. Cytomegalovirus (CMV) can trigger severe disease in immune suppressed individuals, is a frequent cause of congenital birth defects and still represents a major problem in health care. Immunity to acute and persistent CMV infection is intensively studied with respect natural killer (NK) and T cell function. Systemic STAT1 is required for defence against CMV but little is known about the contribution of STAT1 and innate immune cell function. Stat1DLysz mice showed highly efficient Stat1 allele depletion efficiencies in the myeloid compartment. Challenges of Stat1D Lysz with various amounts of MCMV revealed no apparent effect of myeloid STAT1 to survival compared to wildtype mice. However, Stat1DLysz mice were deficient to restrict viral load in various organs and showed a high persistence of MCMV in salivary glands. With respect to early infection and viral spread we investigate cytokine expression and response activities, including cell type specific STAT activation. With respect to MCMV persistence we test the requirement of myeloid STAT1 for the activation of CD4+ and CD8+ T cells. This work was funded by the Austrian Science Fund (FWF) IAI DK W1212 and FWF SFB F28. http://dx.doi.org/10.1016/j.cyto.2013.06.025
23 Complement component C5a permits the co-existence of pathogenic Th17 cells and type I interferon in lupus Partha S. Biswas, Sudesh Pawaria, Kelly Maers, Marc C. Levesque, Department of Medicine, Universit of Pittsburgh, Pittsburgh, PA, USA Systemic lupus erythematosus (SLE) is a type I interferon (IFN-I)-driven autoimmune disorder with exaggerated B and T-helper (Th) cell responses. Th17 cells, a recently identified T-helper cell subset, have been strongly implicated in the pathogenesis of SLE. Since IFN-I suppress the generation and expansion of Th17 cells in an IL-27-dependent manner, it is unclear how pathogenic Th17 cells are generated in SLE in the presence of an environment characterized by high IFN-I levels. Here, we show that activation of complement5a–receptor (C5aR) on macrophages blocks IFN-I-mediated IL-27 production, thus permitting the development of Th17 cells. C5aR activation on IFN-I-responsive macrophages inhibits IRF-1-mediated transactivation of IL-27 gene expression via the PI3K/Akt pathway. Consistently, C5aR-deficient mice exhibited increased IL-27 expression and fewer Th17 cells and consequently reduced lupus nephritis in comparison to wild type mice. In support of these findings in mice, we found that serum from SLE patients inhibited IFN-Iinduced IL-27 production from macrophages. Moreover, the level of serum C5a correlated with Th17 frequency in peripheral blood. Collectively, these data indicate an essential role for C5a in the generation of pathogenic Th17 responses in SLE. Thus therapeutic strategies to block C5aR activation may be beneficial for controlling pathogenic Th17-mediated inflammation in SLE. Research Supported by Dept. of Medicine, University of Pittsburgh, PA
http://dx.doi.org/10.1016/j.cyto.2013.06.026
24 STING, not type I IFN signaling, is required for the mucosal adjuvant activity of Cyclic-di-GMP in vivo Steven Blaaubober, Vincent Gabrielle, Lei Jin, Center for Immunology and Microbial Disease, Albany Medical College, Albany, NY, USA Mucosal vaccination prevents the onset of disease, blocks early colonization, and reduces the risk of horizontal transmission. Currently, there is no vaccine formulation containing a mucosal adjuvant approved for human use. Cyclic-di-GMP (CDG) is a small molecule found only in bacteria, regulating bacterial growth and virulence. CDG exhibits strong immunostimulatory properties in mammalian cells. Karaolis et al. first discovered that CDG activates human immature Dendritic cells (DC), and enhancing DC’s ability to stimulate T cells. Importantly, intranasal co-immunization of CDG and antigens protects host from pathogen infections, including H5N1 influenza, and Streptococcus pneumonia infections in mice. Thus, CDG is a promising mucosal adjuvant candidate. The mechanism by which CDG acts as a mucosal adjuvant is unknown. STING (stimulator of interferon genes) has been identified as a ligand for CDG in mammalian cells activating TBK1-IRF3-type I IFN signaling. Here, using a STING-/- mouse (Tmem173), we investigated the in vivo role of STING in the mucosal adjuvant activity of CDG. We found that STING-/- mice fail to generate antigen-specific antibody response after intranasal immunization of an antigen and CDG. Furthermore, the productions of Th1/Th2/Th17 cytokines, proinflammatory cytokines and type I IFN are all missing in CDG immunized STING-/- mice. Surprisingly, we found that type I IFN signaling is not required for the mucosal adjuvant activity of CDG because IFNAR1-/- mice have the same antigen-specific antibody response as the wild-type mice. We further found that CDG activates STING-dependent but TBK1-type I IFN signaling-independent proinflammatory cytokine production in macrophages and DCs. This is distinct from STING-mediated DNA vaccine adjuvant activity, which requires type I IFN signaling. Thus, our results establish an essential role of STING in mediating the mucosal adjuvant activity of CDG in vivo and reveal novel TBK1-type I IFN stimulation-independent function of STING. http://dx.doi.org/10.1016/j.cyto.2013.06.027
25 STAT2 and IRF9-dependent IFN-I signaling restores ISRE-mediated transcription and anti-viral activity independent of STAT1 Katarzyna Błaszczyk a, Adam Olejnik a, Stefan Chmielewski a, Kaja Kostyrko a, Joanna Wesoly b, Chien-Kuo Lee c, Hans A.R. Bluyssen a, a Department of Human Molecular Genetics and Institute of Molecular Biology and Biotechnology, Faculty of Biology, Adam Mickiewicz University, Poznan, Poland, b Laboratory of High Throughput Technologies,
Abstract / Cytokine 63 (2013) 243–314
cells and the increase in the ratio Teff/Treg in the tumor environment. In conclusion, the favorable pharmacokinetic/pharmacodynamic profile of IL-15 heterodimer allows for lower dose, simple s.c. delivery, and lowers the possibility of toxicity due to cytokine spike. Preclinical cancer studies suggested that IL-15 heterodimer-based immunotherapy favors the development of anti-tumor responses by favoring cytotoxic over regulatory cells. http://dx.doi.org/10.1016/j.cyto.2013.06.023
21 Interferon-mediated potentiation of SMAC mimetic compound cytotoxicity by oncolytic virotherapy Shawn T. Beug a, Vera A. Tang a, Herman C. Cheung a, Fabrice Le Boeuf b, John C. Bell b, Eric C. LaCasse a, Robert G. Korneluk a, a Apoptosis Research Centre, Children’s Hospital of Eastern Ontario Research Institute, Ottawa, Ont., Canada, b Centre for Innovative Cancer Therapeutics, Ottawa Hospital Research Institute, Ottawa, Ont., Canada Smac mimetic compounds (SMCs) are a novel class of small molecule antagonists that target the inhibitor of apoptosis (IAP) proteins to induce cancer cell death via cytokine activation of caspase-8 death pathways. Oncolytic viruses (OVs) selectively target cancer cells for death via multiple mechanisms, including oncolysis and the induction of anti-tumor immunity. We reasoned that oncolytic viruses such as the rhabdovirus VSVD51 will provide a proinflammatory environment that would augment SMC-induced cancer cell death. We observed that SMC and VSVD51 synergy was specific to cancer cells and this combination increased the potency of cancer cell death up to 10,000-fold compared to stand-alone VSVD51 therapy. The combination resulted in delayed tumor growth and lead to durable cures in mouse models of cancer. Furthermore, several non-viral immunostimulatory agents, such as the viral RNA mimic poly (I:C) or the bacterial DNA mimic CpG, elicit similar responses when combined with SMC treatment. The combination leads to bystander cancer cell death via the production of type 1 interferon-dependent production of proinflammatory cytokines, such as TNFa and TRAIL, which sensitizes cancer cells to SMC-mediated caspase-8-dependent apoptosis. We thus demonstrate the novel combination of targeted immunotherapies for the treatment of cancer, which on their own are limited by various efficacy hurdles, yet when combined, the combined treatment results in strong synergy to cause cancer cell death. http://dx.doi.org/10.1016/j.cyto.2013.06.024
22 The role of STAT1 in macrophages/neutrophils in the response to murine cytomegalovirus infection Mario Biaggio a, Caroline Lassnig a,b, Ursula Reichart a,b, Rita Rom a, Astrid Krmpotic c, Stipan Jonjicì c, Birgit Strobl a, Mathias Müller a,b, a Institute of Animal Breeding and Genetics, University of Veterinary Medicine, Vienna, Austria, b Biomodels Austria, University of Veterinary Medicine, Vienna, Austria, c Department of Histology and Embryology, Faculty of Medicine, University of Rijeka, Croatia Signal transducers and activators of transcription (STAT) 1 is a key transcription factor involved in innate and adaptive immune responses by governing the responses to interferons (IFNs) type I, II and III. In order to elucidate the cell type specific contributions of STAT1 to the host response to viral infections our lab generated conditional Stat1 mice. Cytomegalovirus (CMV) can trigger severe disease in immune suppressed individuals, is a frequent cause of congenital birth defects and still represents a major problem in health care. Immunity to acute and persistent CMV infection is intensively studied with respect natural killer (NK) and T cell function. Systemic STAT1 is required for defence against CMV but little is known about the contribution of STAT1 and innate immune cell function. Stat1DLysz mice showed highly efficient Stat1 allele depletion efficiencies in the myeloid compartment. Challenges of Stat1D Lysz with various amounts of MCMV revealed no apparent effect of myeloid STAT1 to survival compared to wildtype mice. However, Stat1DLysz mice were deficient to restrict viral load in various organs and showed a high persistence of MCMV in salivary glands. With respect to early infection and viral spread we investigate cytokine expression and response activities, including cell type specific STAT activation. With respect to MCMV persistence we test the requirement of myeloid STAT1 for the activation of CD4+ and CD8+ T cells. This work was funded by the Austrian Science Fund (FWF) IAI DK W1212 and FWF SFB F28. http://dx.doi.org/10.1016/j.cyto.2013.06.025
23 Complement component C5a permits the co-existence of pathogenic Th17 cells and type I interferon in lupus Partha S. Biswas, Sudesh Pawaria, Kelly Maers, Marc C. Levesque, Department of Medicine, Universit of Pittsburgh, Pittsburgh, PA, USA Systemic lupus erythematosus (SLE) is a type I interferon (IFN-I)-driven autoimmune disorder with exaggerated B and T-helper (Th) cell responses. Th17 cells, a recently identified T-helper cell subset, have been strongly implicated in the pathogenesis of SLE. Since IFN-I suppress the generation and expansion of Th17 cells in an IL-27-dependent manner, it is unclear how pathogenic Th17 cells are generated in SLE in the presence of an environment characterized by high IFN-I levels. Here, we show that activation of complement5a–receptor (C5aR) on macrophages blocks IFN-I-mediated IL-27 production, thus permitting the development of Th17 cells. C5aR activation on IFN-I-responsive macrophages inhibits IRF-1-mediated transactivation of IL-27 gene expression via the PI3K/Akt pathway. Consistently, C5aR-deficient mice exhibited increased IL-27 expression and fewer Th17 cells and consequently reduced lupus nephritis in comparison to wild type mice. In support of these findings in mice, we found that serum from SLE patients inhibited IFN-Iinduced IL-27 production from macrophages. Moreover, the level of serum C5a correlated with Th17 frequency in peripheral blood. Collectively, these data indicate an essential role for C5a in the generation of pathogenic Th17 responses in SLE. Thus therapeutic strategies to block C5aR activation may be beneficial for controlling pathogenic Th17-mediated inflammation in SLE. Research Supported by Dept. of Medicine, University of Pittsburgh, PA
http://dx.doi.org/10.1016/j.cyto.2013.06.026
24 STING, not type I IFN signaling, is required for the mucosal adjuvant activity of Cyclic-di-GMP in vivo Steven Blaaubober, Vincent Gabrielle, Lei Jin, Center for Immunology and Microbial Disease, Albany Medical College, Albany, NY, USA Mucosal vaccination prevents the onset of disease, blocks early colonization, and reduces the risk of horizontal transmission. Currently, there is no vaccine formulation containing a mucosal adjuvant approved for human use. Cyclic-di-GMP (CDG) is a small molecule found only in bacteria, regulating bacterial growth and virulence. CDG exhibits strong immunostimulatory properties in mammalian cells. Karaolis et al. first discovered that CDG activates human immature Dendritic cells (DC), and enhancing DC’s ability to stimulate T cells. Importantly, intranasal co-immunization of CDG and antigens protects host from pathogen infections, including H5N1 influenza, and Streptococcus pneumonia infections in mice. Thus, CDG is a promising mucosal adjuvant candidate. The mechanism by which CDG acts as a mucosal adjuvant is unknown. STING (stimulator of interferon genes) has been identified as a ligand for CDG in mammalian cells activating TBK1-IRF3-type I IFN signaling. Here, using a STING-/- mouse (Tmem173), we investigated the in vivo role of STING in the mucosal adjuvant activity of CDG. We found that STING-/- mice fail to generate antigen-specific antibody response after intranasal immunization of an antigen and CDG. Furthermore, the productions of Th1/Th2/Th17 cytokines, proinflammatory cytokines and type I IFN are all missing in CDG immunized STING-/- mice. Surprisingly, we found that type I IFN signaling is not required for the mucosal adjuvant activity of CDG because IFNAR1-/- mice have the same antigen-specific antibody response as the wild-type mice. We further found that CDG activates STING-dependent but TBK1-type I IFN signaling-independent proinflammatory cytokine production in macrophages and DCs. This is distinct from STING-mediated DNA vaccine adjuvant activity, which requires type I IFN signaling. Thus, our results establish an essential role of STING in mediating the mucosal adjuvant activity of CDG in vivo and reveal novel TBK1-type I IFN stimulation-independent function of STING. http://dx.doi.org/10.1016/j.cyto.2013.06.027
25 STAT2 and IRF9-dependent IFN-I signaling restores ISRE-mediated transcription and anti-viral activity independent of STAT1 Katarzyna Błaszczyk a, Adam Olejnik a, Stefan Chmielewski a, Kaja Kostyrko a, Joanna Wesoly b, Chien-Kuo Lee c, Hans A.R. Bluyssen a, a Department of Human Molecular Genetics and Institute of Molecular Biology and Biotechnology, Faculty of Biology, Adam Mickiewicz University, Poznan, Poland, b Laboratory of High Throughput Technologies,