- Email: [email protected]
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Abstract / Cytokine 70 (2014) 28–79
Results: Ifnar1 and Ifnar2 knockout mice differed markedly in their susceptibility to influenza virus infection; Ifnar2 / > Infar1 / > wildtype. By day 7 post-infection, Ifnar2 / mice displayed evidence of severe disease and were therefore euthanised. We observed no striking differences in viral loads between Ifnar1 / and Ifnar2 / mice. Severe disease displayed by Ifnar2 / mice was associated with increased neutrophil infiltration and elevated levels of IL-1b in the airways and blood. Furthermore, mice lacking Ifnar2 but not Ifnar1, displayed evidence of pulmonary edema and vascular leakage. Conclusions: Overall, our data demonstrates that Infar1 and Infar2 play distinct roles in host defence during influenza virus infection. In particular, Ifnar2 plays an important role in controlling the production of IL-1b and the induction of severe disease. Understanding further this role of Ifnar2 and how type I IFNs control inappropriate or damaging immune responses is of significance to reduce mortality and morbidity associated with influenza. http://dx.doi.org/10.1016/j.cyto.2014.07.188
182 The leukocyte immunoglobulin-like receptor B4 differentially regulates FccRIdependent TNF production via its three intracellular immunoreceptor tyrosinebased inhibitory signalling motifs (ITIMs) Nicodemus Tedla, Meijeong Park, University New South Wales, Sydney, NSW, Australia The leukocyte immunoglobulin-like receptor (LILR) B4 belongs to a family of activating and inhibitory cell surface receptors that have been increasingly recognised as key regulators of the threshold and amplitude of leukocyte activation. LILRB4 which is highly expressed on monocytes and macrophages transduces inhibitory signals through its three intracellular immunoreceptor tyrosine-based inhibitory motifs (ITIMs). Clinically, increased expression is associated with better clinical outcome in renal transplant recipients contrasted with impaired immune effector cell functions in HIV infection. However, mechanisms of how LILRB4 regulates monocyte activation and the exact contribution of the three ITIMs for its immune-regulatory functions are poorly characterised. In this study we show that co-ligation of LILRB4 with FccRI on THP-1 cells and primary monocytes markedly inhibited the production of TNF, a key pro-inflammatory cytokine. Cross-linking of FccRI on THP-1 cells increased phosphorylation key protein tyrosine kinase signaling molecules and an adaptor protein that targets protein tyrosine kinases for degradation (c-Cbl). Co-ligation of LILRB4 and FccRI considerably reduced FccRI-mediated phosphorylation of these signaling molecules and terminated cellular activation by recruiting Src homology region 2 domain-containing phosphatase-1 (SHP-1)-like molecule. Importantly, using site-targeted mutagenesis we found for the first time that the middle (Y412) and distal (Y442) tyrosine residues in its ITIM motifs were critical for the inhibitory functions of LILRB4 and Y412 alone was sufficient for maximal inhibition. By contrast, the proximal tyrosine residue at Y360 showed unexpectedly enhanced FccRI-mediated TNF production. Collectively, our results suggest that LILRB4 is a potent immune regulatory receptor in which combined contribution of its multiple ITIMs may fine tune innate immune responses.
type I interferons response in PBMCs from people with obesity after TLR stimulation. Interestingly, SOCS3 siRNA silencing in non-obese volunteers did not increase type I interferons response. The expression of SOCS3 induced by leptin and endotoxin also inhibited type I interferons response after TLR-3 and TLR-7 stimulation in PBMCs from healthy donors and in U937 cells. Conclusions: Our results show that SOCS3 overexpression inhibits type I interferons response in PBMCs from people with obesity. We also demonstrate that high dose leptin and endotoxin induces SOCS3 expression and inhibits type I interferons in PBMCs from healthy volunteers and in monocytes U937. These results help to explain the aberrant immune response observed in people with obesity during viral infections. http://dx.doi.org/10.1016/j.cyto.2014.07.190
184 Glucocorticosteroids dampen anti-viral defences and enhance the replication of respiratory viruses Belinda J. Thomas 1,2, Rebecca A. Piganis 2, Paul J. Hertzog 2, Michelle D. Tate 2, Philip G. Bardin 1,2, 1 Monash Lung and Sleep, Monash Medical Centre, Melbourne, Vic, Australia, 2 MIMR-PHI Institute, Clayton, VIC, Australia Aims: Viral upper respiratory tract infections caused by rhinovirus (RV) and influenza A virus (IAV) are associated with 50–80% of asthma exacerbations. Inhaled glucocorticosteroids (GCS) are routinely used to relieve asthma symptoms, and while it has been widely reported that GCS reduce inflammation associated with viral infection, the effects of GCS on viral replication has not been comprehensively examined. Methods: Using primary human airway cells and a mouse model of IAV infection we examined the effect of GCS on the production of inflammatory mediators, anti-viral responses and viral replication following RV and/or IAV infection. Results: GCS pre-treatment of epithelial cells significantly reduced IL-8, IL-6 and IP10 production following RV and IAV infection, compared to untreated controls. The expression of anti-viral genes, including ISG56, ISG15, viperin and MxA, were significantly reduced in epithelial cells, fibroblasts and macrophages pre-treated with GCS prior to infection with RV or IAV. The impairment of anti-viral responses observed following GCS pre-treatment was associated with enhanced RV and IAV replication in both epithelial cells and fibroblasts. GCS pre-treatment of mice prior to IAV infection was associated with significant weight loss and severe disease progression compared with untreated mice. Viral loads were significantly elevated in both the lungs and nasal tissues of the GCS treated mice, and there were significantly elevated levels of IL-6, MCP-1 and KC in BAL fluid. Interferon and anti-viral ISG expression were significantly reduced in GCS treated mice. Treatment with recombinant interferon after infection delayed the onset of disease and improved survival. Conclusions: Our data indicate that GCS at therapeutic doses enhanced virus replication through the suppression of innate anti-viral responses. Interferon treatment improved disease severity in the mouse model, suggesting this may be a viable therapeutic option to counteract the adverse effects of GCS treatment on viral replication and innate immune suppression. http://dx.doi.org/10.1016/j.cyto.2014.07.191
http://dx.doi.org/10.1016/j.cyto.2014.07.189
183 Type I interferons response and expression of supressor of cytokine signalling in obesity Elí Terán-Cabanillas, Jesús Hernández, Nutrition, Centro de Investigación en Alimentación y Desarrollo, Hermosillo, Sonora, Mexico Obesity is a pro-inflammatory state related to impaired immune response. We have previously shown that people with obesity present a diminished type I interferon response in PBMCs after TLR stimulation and during influenza A/H1N1 infection. This was accompanied with high expression of suppressor of cytokine signaling-3 (SOCS3) but not SOCS1. SOCS3 is key regulator of type I interferons, but also of leptin and pro-inflammatory cytokines, which are elevated in obesity, and its overexpression is related to insulin and leptin resistance. Now, We hypothesize that high expression of SOCS3 inhibits type I interferons in people with obesity. Furthermore, We analyze the effect of leptin and endotoxin induced inflammation in SOCS expression and the type I interferons response after stimulation with TLR-3 and TLR-7 ligands in PMBCs and U937 cells. Methods: Cytokines were measured by qRT-PCR and/or by ELISA in PBMCs and U937 cells, SOCS1 and SOCS3 expression was measured by qRT-PCR and Western blot. To evaluate the role of SOCS3 and SOCS1 in the inhibition of type I interferons in people with obesity, genes were silenced by siRNA especific for SOCS1 and SOCS3. Results: SOCS3 but not SOCS1 gen silencing by siRNA increased
185 Dealing with ‘Cytokine Storms’: A cultural and linguistic translation of warm factor epidemics wenbing as seasonal viral influenza epidemics in Australia Rey Tiquia, School of Historical and Philosophical Studies (SHAPS), University of Melbourne, Melbourne, VIC, Australia Aim: To demonstrate that clinical space can be used as a platform in translating the ‘language’ of traditional Chinese medicine (TCM) into the ‘language’ of Western biomedical practice. This is in line with a new understanding of science as a knowledge system characterized by locatedness and situatedness. Western biomedicince and TCM are both generated in located and situated clinical practices. Hence, this new understanding of science has a bearing on the relationship between the two medical traditions. Method: A case study on the use of chronoacupuncture in treating seasonal viral influenza or chun wen (spring season ‘warm factor disorder’) in Melbourne will be presented showing how through the medium of clinical practice the ‘language’ of TCM can be expressed in the ‘language’ of biomedical practice. Through this case study, an incubating qi will be seen as an expression of the natural yin and yang order of the flow and metamorphoses of life embedded in specific time and place i.e. the life of the influenza virus embedded within the human body. Using this method of treatment, the symptoms of a ‘cytokine storm’ or a ‘flaring up’ of an incubating qi in spring can be clinically dealt with effectively.
Abstract / Cytokine 70 (2014) 28–79
Results: Ifnar1 and Ifnar2 knockout mice differed markedly in their susceptibility to influenza virus infection; Ifnar2 / > Infar1 / > wildtype. By day 7 post-infection, Ifnar2 / mice displayed evidence of severe disease and were therefore euthanised. We observed no striking differences in viral loads between Ifnar1 / and Ifnar2 / mice. Severe disease displayed by Ifnar2 / mice was associated with increased neutrophil infiltration and elevated levels of IL-1b in the airways and blood. Furthermore, mice lacking Ifnar2 but not Ifnar1, displayed evidence of pulmonary edema and vascular leakage. Conclusions: Overall, our data demonstrates that Infar1 and Infar2 play distinct roles in host defence during influenza virus infection. In particular, Ifnar2 plays an important role in controlling the production of IL-1b and the induction of severe disease. Understanding further this role of Ifnar2 and how type I IFNs control inappropriate or damaging immune responses is of significance to reduce mortality and morbidity associated with influenza. http://dx.doi.org/10.1016/j.cyto.2014.07.188
182 The leukocyte immunoglobulin-like receptor B4 differentially regulates FccRIdependent TNF production via its three intracellular immunoreceptor tyrosinebased inhibitory signalling motifs (ITIMs) Nicodemus Tedla, Meijeong Park, University New South Wales, Sydney, NSW, Australia The leukocyte immunoglobulin-like receptor (LILR) B4 belongs to a family of activating and inhibitory cell surface receptors that have been increasingly recognised as key regulators of the threshold and amplitude of leukocyte activation. LILRB4 which is highly expressed on monocytes and macrophages transduces inhibitory signals through its three intracellular immunoreceptor tyrosine-based inhibitory motifs (ITIMs). Clinically, increased expression is associated with better clinical outcome in renal transplant recipients contrasted with impaired immune effector cell functions in HIV infection. However, mechanisms of how LILRB4 regulates monocyte activation and the exact contribution of the three ITIMs for its immune-regulatory functions are poorly characterised. In this study we show that co-ligation of LILRB4 with FccRI on THP-1 cells and primary monocytes markedly inhibited the production of TNF, a key pro-inflammatory cytokine. Cross-linking of FccRI on THP-1 cells increased phosphorylation key protein tyrosine kinase signaling molecules and an adaptor protein that targets protein tyrosine kinases for degradation (c-Cbl). Co-ligation of LILRB4 and FccRI considerably reduced FccRI-mediated phosphorylation of these signaling molecules and terminated cellular activation by recruiting Src homology region 2 domain-containing phosphatase-1 (SHP-1)-like molecule. Importantly, using site-targeted mutagenesis we found for the first time that the middle (Y412) and distal (Y442) tyrosine residues in its ITIM motifs were critical for the inhibitory functions of LILRB4 and Y412 alone was sufficient for maximal inhibition. By contrast, the proximal tyrosine residue at Y360 showed unexpectedly enhanced FccRI-mediated TNF production. Collectively, our results suggest that LILRB4 is a potent immune regulatory receptor in which combined contribution of its multiple ITIMs may fine tune innate immune responses.
type I interferons response in PBMCs from people with obesity after TLR stimulation. Interestingly, SOCS3 siRNA silencing in non-obese volunteers did not increase type I interferons response. The expression of SOCS3 induced by leptin and endotoxin also inhibited type I interferons response after TLR-3 and TLR-7 stimulation in PBMCs from healthy donors and in U937 cells. Conclusions: Our results show that SOCS3 overexpression inhibits type I interferons response in PBMCs from people with obesity. We also demonstrate that high dose leptin and endotoxin induces SOCS3 expression and inhibits type I interferons in PBMCs from healthy volunteers and in monocytes U937. These results help to explain the aberrant immune response observed in people with obesity during viral infections. http://dx.doi.org/10.1016/j.cyto.2014.07.190
184 Glucocorticosteroids dampen anti-viral defences and enhance the replication of respiratory viruses Belinda J. Thomas 1,2, Rebecca A. Piganis 2, Paul J. Hertzog 2, Michelle D. Tate 2, Philip G. Bardin 1,2, 1 Monash Lung and Sleep, Monash Medical Centre, Melbourne, Vic, Australia, 2 MIMR-PHI Institute, Clayton, VIC, Australia Aims: Viral upper respiratory tract infections caused by rhinovirus (RV) and influenza A virus (IAV) are associated with 50–80% of asthma exacerbations. Inhaled glucocorticosteroids (GCS) are routinely used to relieve asthma symptoms, and while it has been widely reported that GCS reduce inflammation associated with viral infection, the effects of GCS on viral replication has not been comprehensively examined. Methods: Using primary human airway cells and a mouse model of IAV infection we examined the effect of GCS on the production of inflammatory mediators, anti-viral responses and viral replication following RV and/or IAV infection. Results: GCS pre-treatment of epithelial cells significantly reduced IL-8, IL-6 and IP10 production following RV and IAV infection, compared to untreated controls. The expression of anti-viral genes, including ISG56, ISG15, viperin and MxA, were significantly reduced in epithelial cells, fibroblasts and macrophages pre-treated with GCS prior to infection with RV or IAV. The impairment of anti-viral responses observed following GCS pre-treatment was associated with enhanced RV and IAV replication in both epithelial cells and fibroblasts. GCS pre-treatment of mice prior to IAV infection was associated with significant weight loss and severe disease progression compared with untreated mice. Viral loads were significantly elevated in both the lungs and nasal tissues of the GCS treated mice, and there were significantly elevated levels of IL-6, MCP-1 and KC in BAL fluid. Interferon and anti-viral ISG expression were significantly reduced in GCS treated mice. Treatment with recombinant interferon after infection delayed the onset of disease and improved survival. Conclusions: Our data indicate that GCS at therapeutic doses enhanced virus replication through the suppression of innate anti-viral responses. Interferon treatment improved disease severity in the mouse model, suggesting this may be a viable therapeutic option to counteract the adverse effects of GCS treatment on viral replication and innate immune suppression. http://dx.doi.org/10.1016/j.cyto.2014.07.191
http://dx.doi.org/10.1016/j.cyto.2014.07.189
183 Type I interferons response and expression of supressor of cytokine signalling in obesity Elí Terán-Cabanillas, Jesús Hernández, Nutrition, Centro de Investigación en Alimentación y Desarrollo, Hermosillo, Sonora, Mexico Obesity is a pro-inflammatory state related to impaired immune response. We have previously shown that people with obesity present a diminished type I interferon response in PBMCs after TLR stimulation and during influenza A/H1N1 infection. This was accompanied with high expression of suppressor of cytokine signaling-3 (SOCS3) but not SOCS1. SOCS3 is key regulator of type I interferons, but also of leptin and pro-inflammatory cytokines, which are elevated in obesity, and its overexpression is related to insulin and leptin resistance. Now, We hypothesize that high expression of SOCS3 inhibits type I interferons in people with obesity. Furthermore, We analyze the effect of leptin and endotoxin induced inflammation in SOCS expression and the type I interferons response after stimulation with TLR-3 and TLR-7 ligands in PMBCs and U937 cells. Methods: Cytokines were measured by qRT-PCR and/or by ELISA in PBMCs and U937 cells, SOCS1 and SOCS3 expression was measured by qRT-PCR and Western blot. To evaluate the role of SOCS3 and SOCS1 in the inhibition of type I interferons in people with obesity, genes were silenced by siRNA especific for SOCS1 and SOCS3. Results: SOCS3 but not SOCS1 gen silencing by siRNA increased
185 Dealing with ‘Cytokine Storms’: A cultural and linguistic translation of warm factor epidemics wenbing as seasonal viral influenza epidemics in Australia Rey Tiquia, School of Historical and Philosophical Studies (SHAPS), University of Melbourne, Melbourne, VIC, Australia Aim: To demonstrate that clinical space can be used as a platform in translating the ‘language’ of traditional Chinese medicine (TCM) into the ‘language’ of Western biomedical practice. This is in line with a new understanding of science as a knowledge system characterized by locatedness and situatedness. Western biomedicince and TCM are both generated in located and situated clinical practices. Hence, this new understanding of science has a bearing on the relationship between the two medical traditions. Method: A case study on the use of chronoacupuncture in treating seasonal viral influenza or chun wen (spring season ‘warm factor disorder’) in Melbourne will be presented showing how through the medium of clinical practice the ‘language’ of TCM can be expressed in the ‘language’ of biomedical practice. Through this case study, an incubating qi will be seen as an expression of the natural yin and yang order of the flow and metamorphoses of life embedded in specific time and place i.e. the life of the influenza virus embedded within the human body. Using this method of treatment, the symptoms of a ‘cytokine storm’ or a ‘flaring up’ of an incubating qi in spring can be clinically dealt with effectively.