- Email: [email protected]
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Abstract / Cytokine 70 (2014) 28–79
absence of TCR engagement to produce IL-17A and IL-17F and express the master transcription factor, RORct and the integrin, MCAM. Furthermore, Vc4bþ T cells, together with conventional Vc4dþ T cells were found in the brains of mice with EAE. Vc4bþ T cells were also found in the brains TCRd= mice with EAE. Although, the course of EAE is somewhat reduced in TCRd= compared with wild type (WT) mice, depletion of Vc4+ T cells from TCRd= as well as WT mice significantly attenuated clinical disease and weight loss in mice with EAE. Anti-Vc4 treated mice with EAE had a significantly reduced frequency of infiltrating IL-17+, IFN-cþ and IFN-cþ IL17+ CD4+ T cells into CNS. Furthermore, in the adoptive transfer model of EAE, depletion of Vc4+ cells from lymph node and spleen cells from TCRd= mice significantly impair their ability to induce EAE, with a reduction in inflammatory T cells infiltrating the CNS. Our study has identified a novel cb T cell subset, Vc4b T cells, which together with conventional Vc4d T cells, play a critical role in the pathogenesis of EAE through innate IL-17 production, necessary to enhance Th17 and Th1 activation and migration into the CNS to mediate inflammation and autoimmunity.
IFN innate response, as well as ex vivo type I IFN production associated with strong induction of antiviral PRRs (RIG-I, MDA5), transcription factors (IRF-3, IRF-7), and IFN-dependent (PKR, Oas1, Mx) and independent ISGs (ISG49, ISG54, ISG56) by alternative activation of IRF3 and NF-jB in myeloid-derived DCs and macrophages, as compared to TLR3/ myeloid-derived cells which were more permissive to viral replication through impaired type I IFN innate response. TLR4 ablation also appeared to mount an enhanced type I IFN innate and humoral, CD4 + and CD8 + T cell responses, which were mediated by altered immune cell populations (increased number of plasmacytoid DCs and NK cells, reduced CD11b + Ly-6Chigh monocytes) and CD4 + Foxp3 + Treg number in lymphoid tissue. Thus, potent type I IFN innate and adaptive immune responses in the absence of TLR4 were closely coupled with reduced JE lethality. Collectively, these results suggest that a balanced triggering of TLR signal array by viral components during JE progression could be responsible for determining disease outcome through regulating negative and positive factors. http://dx.doi.org/10.1016/j.cyto.2014.07.050
http://dx.doi.org/10.1016/j.cyto.2014.07.048
42 Apics deficiency reveals a role for a long noncoding RNA in dendritic cell function and autoimmunity Colleen M. Elso 1, Michelle P. Ashton 1,2, Leanne Mackin 1, Edward Chu 1,2, Natalie L. Payne 3, Sharon Ford 4,5, Iris K.L. Tan 6, Anthony T. Papenfuss 7, A. Richard Kitching 4,5, Shaun Summers 4,5, Claude Bernard 3, Ken Shortman 6,7, Grant Morahan 8, Meredith O’Keeffe 7, Thomas C. Brodnicki 1, 1 St Vincent’s Institute, Fitzroy, VIC, Australia, 2 Department of Medicine, University of Melbourne, Parkville, VIC, Australia, 3 Monash Immunology and Stem Cell Laboratories, Monash University, Clayton, VIC, Australia, 4 Centre for Inflammatory Diseases, Department of Medicine, Monash University, Clayton, VIC, Australia, 5 Department of Nephrology, Monash Medical Centre, Clayton, VIC, Australia, 6 The Walter and Eliza Hall Institute, Parkville, VIC, Australia, 7 Burnet Institute, Melbourne, VIC, Australia, 8 The Western Australian Institute of Medical Research, Perth, WA, Australia Although in vitro observations indicate that long noncoding RNAs (lncRNAs) regulate innate immune responses, their effect upon immune tolerance in vivo has not been defined. We have identified a novel gene of unknown function for which sequence variation is associated with autoimmune diabetes in the nonobese diabetic (NOD) mouse strain. Bioinformatics and expression analyses indicate this gene encodes a lncRNA that is induced by Toll-like receptor (TLR) activation, localises to the nucleus and cytoplasm of dendritic cells, and binds to proteins within these cellular compartments. Moreover, sequence variation for this gene is associated with altered TLR-mediated cytokine production. Hence this gene was named Apics for Attenuator of Pattern recognition receptor-Induced Cytokine Secretion. To further investigate the function of this lncRNA, we established a C57BL/6 (B6) knockout mouse strain for Apics and discovered that Apics-deficient dendritic cells exhibit enhanced TLR-mediated cytokine production. Apics-deficient B6 mice also exhibit increased susceptibility to autoimmunity in two disease models: experimental autoimmune encephalomyelitis and experimental anti-neutrophil cytoplasmic antibodyassociated vasculitis. Our study suggests that lncRNAs, such as Apics, can serve as TLR-inducible repressors that regulate the magnitude of innate immune responses to reduce the risk for developing autoimmune disease.
44 CD11chigh PDCA-1int/low dendritic cells regulates the differentiation of CD11b+Ly6Chigh monocytes and permeability of blood–brain barrier to galvanize the immune privileged CNS to neuroinflammation Seong Kug Eo, Jin Hyoung Kim, Jin Young Choi, Seong Bum Kim, Erdenebelig Uyangaa, Ajit Mahadev Patil, Department Of Microbiology, College Of Veterinary Medicine, Chonbuk, Republic of Korea Although there were bunch of results showing the defined role of dendritic cells (DCs) in initiating adaptive host defense, their potential contributions to T-cell independent innate host defense and to subsequent immunopathology in immune privileged central nervous system (CNS) by neurotrophic virus is not completely defined. Notably, the potential roles of DC-monocyte interaction during neuroinflammatory course have not yet been addressed in depth. Here we found that CD11chighPDCA1int/low DCs are essential to control the progression of neuroinflammation in immune privileged CNS upon infection of neurotrophic Japanese encephalitis virus (JEV), using selective ablation of CD11chighPDCA-1int/low DCs without altered CD11c[int]PDCA1high plasmacytoid DC number, as manifested by increased CD11b+Ly-6Chigh monocyte infiltration, blood–brain barrier (BBB) permeability, viral burden, and cytokine expression. More surprisingly, CD11b+Ly-6Chigh monocytes were observed to have deleterious role in viral encephalitis, and selective ablation of CD11chighPDCA-1int/ low DCs provided altered differentiation and function of infiltrated CD11b+Ly6Chighmonocytes in CNS. Subsequently, CD11b+Ly-6Chigh monocytes generated in the absence of CD11chighPDCA-1int/low DCs elicited faster recruitment into inflamed CNS depending on CCR2, and augmented the severity of neuroinflammation caused by JEV infection, as were proved by several experimental models including adoptive transfer of CD11b+Ly-6Chigh monocytes using CCR2 KO and CCR2 KOCD11c-DTR BM chimera. Our data identify for the first time an unrecognized role of CD11chighPDCA1int/low DCs in maintaining the functional homeostasis of inflammatory CD11b+Ly6Chigh monocytes and integrity of BBB, thereby affecting immunopathology in immune privileged CNS by neurotrophic virus. http://dx.doi.org/10.1016/j.cyto.2014.07.051
http://dx.doi.org/10.1016/j.cyto.2014.07.049
43 Distinct dictation of Japanese encephalitis virus-induced neuroinflammation and lethality via triggering TLR3 and TLR4 signal pathways Seong Kug Eo, Young Woo Han, Jin Young Choi, Erdenebelig Uyangaa, Seong Bum Kim, Jin Hyoung Kim, Bum Seok Kim, Department Of Microbiology, College Of Veterinary Medicine, Chonbuk, Republic of Korea Japanese encephalitis (JE) is major emerging neurologic disease caused by JE virus. To date, the impact of TLR molecules on JE progression has not been addressed. Here, we determined whether each TLR modulates JE, using several TLR-deficient mouse strains (TLR2, TLR3, TLR4, TLR7, TLR9). Surprisingly, among the tested TLRdeficient mice there were contrasting results in TLR3/ and TLR4/ mice, i.e. TLR3/ mice were highly susceptible to JE, whereas TLR4/ mice showed enhanced resistance to JE. TLR3 ablation induced severe CNS inflammation characterized by early infiltration of inflammatory CD11b + Ly-6Chigh monocytes along with profoundly increased viral burden, proinflammatory cytokine/chemokine expression as well as BBB permeability. In contrast, TLR4/ mice showed mild CNS inflammation manifested by reduced viral burden, leukocyte infiltration and proinflammatory cytokine expression. Interestingly, TLR4 ablation provided potent in vivo systemic type I
45 Functional restoration of exhausted CD4+ and CD8+ T cells in chronic viral infection by daphnane diterpene ester derived from Daphne genkwa flos buds via negative regulatory Tim-3 molecule Seong Kug Eo, Erdenebelig Uyangaa, Jin Young Choi, Ajit Mahadev Patil, Jin Hyoung Kim, Seong Bum Kim, Department Of Microbiology, College Of Veterinary Medicine, Chonbuk, Republic of Korea CD4+ and CD8+ T-cell exhaustion developed in chronic viral infection has been important issue because exhausted antigen-specific CD4+ and CD8+ T cells showed impaired ability to eradicate persistently infected virus and produce effector cytokines such as IFN-c and TNF-a. Hence, strategies that either restore endogenous exhausted T-cell responses or provide functional CD4+ and CD8+ T cells need to be urgently developed for therapeutics of human chronic infection. In spite of promising development using antibody and cell immunotherapy, there were no attempts to restore exhausted CD4+ and CD8+ T cells using treatment of small bioactive molecules, especially derived from natural resources. Here, using mouse model of chronic infection with lymphocytic choriomeningitis virus (LCMV), we found for the first time that genkwadaphnin isolated from dried flower buds of Daphne genkwa (Thymelaeceae) significantly restored exhausted CD4+ and CD8+ T cells, as corroborated by evidences that genkwadaphnin treatment enhanced functional LCMV-specific CD4+ and CD8+ T cells in both quantitative and qualitative. Repeated administration of lower
Abstract / Cytokine 70 (2014) 28–79
absence of TCR engagement to produce IL-17A and IL-17F and express the master transcription factor, RORct and the integrin, MCAM. Furthermore, Vc4bþ T cells, together with conventional Vc4dþ T cells were found in the brains of mice with EAE. Vc4bþ T cells were also found in the brains TCRd= mice with EAE. Although, the course of EAE is somewhat reduced in TCRd= compared with wild type (WT) mice, depletion of Vc4+ T cells from TCRd= as well as WT mice significantly attenuated clinical disease and weight loss in mice with EAE. Anti-Vc4 treated mice with EAE had a significantly reduced frequency of infiltrating IL-17+, IFN-cþ and IFN-cþ IL17+ CD4+ T cells into CNS. Furthermore, in the adoptive transfer model of EAE, depletion of Vc4+ cells from lymph node and spleen cells from TCRd= mice significantly impair their ability to induce EAE, with a reduction in inflammatory T cells infiltrating the CNS. Our study has identified a novel cb T cell subset, Vc4b T cells, which together with conventional Vc4d T cells, play a critical role in the pathogenesis of EAE through innate IL-17 production, necessary to enhance Th17 and Th1 activation and migration into the CNS to mediate inflammation and autoimmunity.
IFN innate response, as well as ex vivo type I IFN production associated with strong induction of antiviral PRRs (RIG-I, MDA5), transcription factors (IRF-3, IRF-7), and IFN-dependent (PKR, Oas1, Mx) and independent ISGs (ISG49, ISG54, ISG56) by alternative activation of IRF3 and NF-jB in myeloid-derived DCs and macrophages, as compared to TLR3/ myeloid-derived cells which were more permissive to viral replication through impaired type I IFN innate response. TLR4 ablation also appeared to mount an enhanced type I IFN innate and humoral, CD4 + and CD8 + T cell responses, which were mediated by altered immune cell populations (increased number of plasmacytoid DCs and NK cells, reduced CD11b + Ly-6Chigh monocytes) and CD4 + Foxp3 + Treg number in lymphoid tissue. Thus, potent type I IFN innate and adaptive immune responses in the absence of TLR4 were closely coupled with reduced JE lethality. Collectively, these results suggest that a balanced triggering of TLR signal array by viral components during JE progression could be responsible for determining disease outcome through regulating negative and positive factors. http://dx.doi.org/10.1016/j.cyto.2014.07.050
http://dx.doi.org/10.1016/j.cyto.2014.07.048
42 Apics deficiency reveals a role for a long noncoding RNA in dendritic cell function and autoimmunity Colleen M. Elso 1, Michelle P. Ashton 1,2, Leanne Mackin 1, Edward Chu 1,2, Natalie L. Payne 3, Sharon Ford 4,5, Iris K.L. Tan 6, Anthony T. Papenfuss 7, A. Richard Kitching 4,5, Shaun Summers 4,5, Claude Bernard 3, Ken Shortman 6,7, Grant Morahan 8, Meredith O’Keeffe 7, Thomas C. Brodnicki 1, 1 St Vincent’s Institute, Fitzroy, VIC, Australia, 2 Department of Medicine, University of Melbourne, Parkville, VIC, Australia, 3 Monash Immunology and Stem Cell Laboratories, Monash University, Clayton, VIC, Australia, 4 Centre for Inflammatory Diseases, Department of Medicine, Monash University, Clayton, VIC, Australia, 5 Department of Nephrology, Monash Medical Centre, Clayton, VIC, Australia, 6 The Walter and Eliza Hall Institute, Parkville, VIC, Australia, 7 Burnet Institute, Melbourne, VIC, Australia, 8 The Western Australian Institute of Medical Research, Perth, WA, Australia Although in vitro observations indicate that long noncoding RNAs (lncRNAs) regulate innate immune responses, their effect upon immune tolerance in vivo has not been defined. We have identified a novel gene of unknown function for which sequence variation is associated with autoimmune diabetes in the nonobese diabetic (NOD) mouse strain. Bioinformatics and expression analyses indicate this gene encodes a lncRNA that is induced by Toll-like receptor (TLR) activation, localises to the nucleus and cytoplasm of dendritic cells, and binds to proteins within these cellular compartments. Moreover, sequence variation for this gene is associated with altered TLR-mediated cytokine production. Hence this gene was named Apics for Attenuator of Pattern recognition receptor-Induced Cytokine Secretion. To further investigate the function of this lncRNA, we established a C57BL/6 (B6) knockout mouse strain for Apics and discovered that Apics-deficient dendritic cells exhibit enhanced TLR-mediated cytokine production. Apics-deficient B6 mice also exhibit increased susceptibility to autoimmunity in two disease models: experimental autoimmune encephalomyelitis and experimental anti-neutrophil cytoplasmic antibodyassociated vasculitis. Our study suggests that lncRNAs, such as Apics, can serve as TLR-inducible repressors that regulate the magnitude of innate immune responses to reduce the risk for developing autoimmune disease.
44 CD11chigh PDCA-1int/low dendritic cells regulates the differentiation of CD11b+Ly6Chigh monocytes and permeability of blood–brain barrier to galvanize the immune privileged CNS to neuroinflammation Seong Kug Eo, Jin Hyoung Kim, Jin Young Choi, Seong Bum Kim, Erdenebelig Uyangaa, Ajit Mahadev Patil, Department Of Microbiology, College Of Veterinary Medicine, Chonbuk, Republic of Korea Although there were bunch of results showing the defined role of dendritic cells (DCs) in initiating adaptive host defense, their potential contributions to T-cell independent innate host defense and to subsequent immunopathology in immune privileged central nervous system (CNS) by neurotrophic virus is not completely defined. Notably, the potential roles of DC-monocyte interaction during neuroinflammatory course have not yet been addressed in depth. Here we found that CD11chighPDCA1int/low DCs are essential to control the progression of neuroinflammation in immune privileged CNS upon infection of neurotrophic Japanese encephalitis virus (JEV), using selective ablation of CD11chighPDCA-1int/low DCs without altered CD11c[int]PDCA1high plasmacytoid DC number, as manifested by increased CD11b+Ly-6Chigh monocyte infiltration, blood–brain barrier (BBB) permeability, viral burden, and cytokine expression. More surprisingly, CD11b+Ly-6Chigh monocytes were observed to have deleterious role in viral encephalitis, and selective ablation of CD11chighPDCA-1int/ low DCs provided altered differentiation and function of infiltrated CD11b+Ly6Chighmonocytes in CNS. Subsequently, CD11b+Ly-6Chigh monocytes generated in the absence of CD11chighPDCA-1int/low DCs elicited faster recruitment into inflamed CNS depending on CCR2, and augmented the severity of neuroinflammation caused by JEV infection, as were proved by several experimental models including adoptive transfer of CD11b+Ly-6Chigh monocytes using CCR2 KO and CCR2 KOCD11c-DTR BM chimera. Our data identify for the first time an unrecognized role of CD11chighPDCA1int/low DCs in maintaining the functional homeostasis of inflammatory CD11b+Ly6Chigh monocytes and integrity of BBB, thereby affecting immunopathology in immune privileged CNS by neurotrophic virus. http://dx.doi.org/10.1016/j.cyto.2014.07.051
http://dx.doi.org/10.1016/j.cyto.2014.07.049
43 Distinct dictation of Japanese encephalitis virus-induced neuroinflammation and lethality via triggering TLR3 and TLR4 signal pathways Seong Kug Eo, Young Woo Han, Jin Young Choi, Erdenebelig Uyangaa, Seong Bum Kim, Jin Hyoung Kim, Bum Seok Kim, Department Of Microbiology, College Of Veterinary Medicine, Chonbuk, Republic of Korea Japanese encephalitis (JE) is major emerging neurologic disease caused by JE virus. To date, the impact of TLR molecules on JE progression has not been addressed. Here, we determined whether each TLR modulates JE, using several TLR-deficient mouse strains (TLR2, TLR3, TLR4, TLR7, TLR9). Surprisingly, among the tested TLRdeficient mice there were contrasting results in TLR3/ and TLR4/ mice, i.e. TLR3/ mice were highly susceptible to JE, whereas TLR4/ mice showed enhanced resistance to JE. TLR3 ablation induced severe CNS inflammation characterized by early infiltration of inflammatory CD11b + Ly-6Chigh monocytes along with profoundly increased viral burden, proinflammatory cytokine/chemokine expression as well as BBB permeability. In contrast, TLR4/ mice showed mild CNS inflammation manifested by reduced viral burden, leukocyte infiltration and proinflammatory cytokine expression. Interestingly, TLR4 ablation provided potent in vivo systemic type I
45 Functional restoration of exhausted CD4+ and CD8+ T cells in chronic viral infection by daphnane diterpene ester derived from Daphne genkwa flos buds via negative regulatory Tim-3 molecule Seong Kug Eo, Erdenebelig Uyangaa, Jin Young Choi, Ajit Mahadev Patil, Jin Hyoung Kim, Seong Bum Kim, Department Of Microbiology, College Of Veterinary Medicine, Chonbuk, Republic of Korea CD4+ and CD8+ T-cell exhaustion developed in chronic viral infection has been important issue because exhausted antigen-specific CD4+ and CD8+ T cells showed impaired ability to eradicate persistently infected virus and produce effector cytokines such as IFN-c and TNF-a. Hence, strategies that either restore endogenous exhausted T-cell responses or provide functional CD4+ and CD8+ T cells need to be urgently developed for therapeutics of human chronic infection. In spite of promising development using antibody and cell immunotherapy, there were no attempts to restore exhausted CD4+ and CD8+ T cells using treatment of small bioactive molecules, especially derived from natural resources. Here, using mouse model of chronic infection with lymphocytic choriomeningitis virus (LCMV), we found for the first time that genkwadaphnin isolated from dried flower buds of Daphne genkwa (Thymelaeceae) significantly restored exhausted CD4+ and CD8+ T cells, as corroborated by evidences that genkwadaphnin treatment enhanced functional LCMV-specific CD4+ and CD8+ T cells in both quantitative and qualitative. Repeated administration of lower