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PS1-38 Interleukin-6 blockade in CD4 T cells inhibits the development of experimental autoimmune uveitis
Abstracts / Cytokine 52 (2010) 17–34
25
doi:10.1016/j.cyto.2010.07.108
University Graduate School of Dentistry, Sendai, Japan, 2 Division of Orthodontics and Dentofacial Orthopedics, Department of Oral Health and Development Sciences, Tohoku University Graduate School of Dentistry, Sendai, Japan
PS1-37 Aryl hydorcarbon receptor mediates suppression of colon inflamation induced by dextran sulfate sodium Ichino Chinen, Akihiro Kimura, Taisuke Nakahama, Kazuya Masuda, Tadamitsu Kishimoto, Laboratory of Immune Regulation, Graduate school of Frontier Bioscienses, Osaka University, Suita, Osaka, Japan
Objectives: We reported that lipopolysaccharide (LPS) promoted and augmented Ni-allergy in mice, and we thereby established a reproducible murine Ni-allergy model termed ‘Ni(+LPS)-allergy’. Histamine, an inflammatory mediator, also modifies Th1/ Th2 responses. It is widely known that histamine is pooled in mast cells or basophils, but less well known that the histamine-forming enzyme, histidine decarboxylase (HDC), is an inducible enzyme and that IL-1 and microbial substances (including LPS) can induce HDC in various murine tissues, even in mast cell-deficient mice. We found that HDC was induced at the site of Ni(+LPS)-allergy, and that Ni(+LPS)-allergy was induced in mast cell-deficient mice and even in nude mice that lack T-cells, but very weak either in HDC-deficient or in IL-1-deficent mice. Here, we report the roles of IL-1 and histamine in Ni(+LPS)-allergy. Methods: A mixture of NiCl2 and LPS was injected intraperitoneally into mice. Ten days later, NiCl2 was challenged intradermally into ears, and ear thickness was measured. In cell-transfer experiments, spleen cells from sensitized or non-sensitized mice (HDC-KO, histamine receptor H1-KO, IL1(both a and b types)-KO mice, or control wild type mice) were injected intravenously into non-sensitized mice, and 24h later, NiCl2 was challenged. Results: The results obtained from cell-transfer experiments demonstrated that (i) the elicitation step involves stimulation of H1 receptors by histamine newly formed via the induction of HDC, and (ii) IL-1 is involved in both the sensitization and elicitation steps. Conclusions: In murine Ni(+LPS)-allergy, IL-1 may be involved in the sensitization step through a histamine-independent mechanism, and it is likely that histamine, which is produced by an IL-1-dependent HDC induction, may be involved in the elicitation step.
Inflammatory bowel disease (IBD) is a chronic disorder of the gastrointestinal tract. It has become clear that Th1 or Th2, cause worsening symptoms, and moreover that IBDs are characterized by a sustained production of cytokines produced by newly identified Th17 cells. Our group and others have demonstrated that Aryl hydrocarbon receptor (Ahr) participates in Th17 cell differentiation and modulates the Th1/Th2 balance. In this study, we investigated the role of Ahr in the development of dextran sulfate sodium (DSS)-induced colitis. Wild type (WT) and Ahr deficient (KO) mice were given 1% DSS dissolved in drinking water. All Ahr KO mice, but not WT littermates, died within 9 days after DSS administration. Moreover, significant body weight loss in Ahr KO mice was observed following 4 days of DSS administration. Conversely, WT mice receiving 1% DSS showed no loss in body weight. Flow cytometry was used to determine the differentiation of T cells in colonic lamina propria lymphocytes (LPLs) from 1% DSS-treated WT and Ahr KO mice for 5 days. A significant increase in the percentage of Th1 cells was observed in DSS-treated Ahr KO mice compared with DSS-treated WT mice. However, there was no difference in Treg and Th17 cell development in LPLs from DSS-treated WT and Ahr KO mice. Furthermore, we found that pro-inflammatory cytokines including IL-12 were increased, in colon-cultured supernatant of 1%DSS treated Ahr KO mice compared to that of WT mice. It has also been reported that Ahr regulates the production of LPSinduced pro-inflammatory cytokines including IL-12, which induces Th1 cell development. Ahr may inhibit the development of Th1 cells through regulating IL-12 secretion from activated macrophages, which may result in DSS-induced colonic inflammation. We intend to further investigate the mechanistic detail of Ahr function in DSS-induced colitis using macrophage- and T cell- specific Ahr deficient mice.
doi:10.1016/j.cyto.2010.07.111
PS1-40 Stimulation of LY-6G on neutrophils in lipopolysaccharide-primed mice induces platelet-activating factor-mediated anaphylaxis-like shock Yukinori Tanaka, Yasuhiro Nagai, Toshinobu Kuroishi, Yasuo Endo, Shunji Sugawara, Division of Oral Immunology, Department of Oral Biology, Tohoku University Graduate School of Dentistry, Sendai, Japan
doi:10.1016/j.cyto.2010.07.109
PS1-38 Interleukin-6 blockade in CD4 T cells inhibits the development of experimental autoimmune uveitis Hiroshi Haruta 1, Nobuyuki Ohguro 1, Satoshi Hohki 1, Kohji Nishida 1, Minoru Fujimoto 2, Tetsuji Naka 2, 1 Department of Ophthalmology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan, 2 Laboratory for Immune Signal, National Institute of Biomedical Innovation, Ibaraki, Osaka, Japan Purpose: A novel subset of interleukin (IL)-17-producing CD4+ T helper cells (Th17) is thought to play a pivotal role in experimental autoimmune uveitis (EAU) and human uveitis. Recent studies also have indicated that IL-6 promotes Th17 differentiation in vitro and in vivo. The aim of this study is to investigate the role of IL-6 signaling in CD4 T cells in the development of EAU. Methods: EAU was induced by immunization with interphotoreceptor retinoid binding protein (IRBP) in wild type (WT), IL-6 knockout (KO) mice and T cell-specific signal transducer activated signal transducers and activator of transcription 3 (STAT3) KO female mice, which are defective in IL-6 signaling. All experimental mice used were on the C57BL/6 background. The clinical scores of EAU and CD4 T cell differentiation in these mice were analyzed. All mice were treated according to the ARVO Statement on the Use of Animals in Ophthalmic and Vision Research. Results: EAU did not develop in IL-6KO and STAT3KO mice while WT mice displayed severe inflammation. Th17 cell differentiation in cervical lymph nodes on day 10 after immunization also decreased in IL-6KO and STAT3KO compared to WT mice. IRBP specific CD4+CD25+ T regulatory cells were observed to be significantly increased in IL-6KO mice. Conclusion: This study showed that IL-6 and STAT3 in T cells are essential in the development of EAU and for the differentiation of highly pro-inflammatory Th17 cells. The protective effect of defective IL-6 signaling in the development of EAU is associated with the inhibition of differentiation of highly pro-inflammatory Th17 cells and an increase in the frequency of immunosuppressive T regulatory cells. doi:10.1016/j.cyto.2010.07.110
PS1-39 Roles of IL-1 and histamine in nickel (Ni) allergy in mice Masayuki Kinbara 1,2, Yasuhiro Nagai 1, Teruko Takano-Yamamoto 2, Yasuo Endo 1, Shunji Sugawara 1, 1 Division of Oral Immunology, Department of Oral Biology, Tohoku
Two anti-Ly-6G mAbs, RB6-8C5 and 1A8, have been used to deplete neutrophils in mice and to clarify their involvement in immune responses. During the course of experiments on neutrophil depletion, we noticed that intravenous injection of RB68C5 induced anaphylaxis-like shock in mice pretreated with lipopolysaccharide (LPS). We investigated the mechanism and obtained the following results. Signs of shock, such as hypothermia, appeared within a few minutes, and the mice died of shock within 20 min of the RB6-8C5 injection. Similar results were obtained with another anti-Ly-6G mAb, 1A8. Optimal LPS priming required at least 1 h. The priming effects of LPS may be associated with neutrophil accumulation in lung and liver (which began at 1 and 1.5 h after LPS injection, respectively). These neutrophils were localized within pulmonary and hepatic blood vessels. A platelet-activating factor (PAF)-receptor antagonist, CV-3988, prevented the shock. Moreover, a PAF biosynthetic enzyme, lyso-PAF acetyltransferase, was induced in the lungs by RB6-8C5 injection into mice pretreated with LPS. These results suggest that antiLy-6G mAbs may lead to PAF production by LPS-primed neutrophils located within pulmonary and hepatic blood vessels, resulting in rapid, lethal anaphylaxis-like shock. doi:10.1016/j.cyto.2010.07.112
PS1-41 Aryl hydrocarbon receptor deficiency significantly suppresses collagen induced arthritis in mice Nakahama T, Kimura A, Chinen I, Masuda K, Nguyen N, Kishimoto T, Laboratory of Immune Regulation, Graduate school of Frontier Bioscienses, Osaka University, Suita, Osaka, Japan Aryl hydrocarbon receptor (Ahr), as known a dioxin receptor, is a ligand-dependent transcription factor. It has been reported that Ahr plays an important role in the immune system. Our recent study demonstrated that Ahr participates in Th17 cell development and regulates inflammatory responses by lipopolysaccharide (LPS) in macrophage. In this study, we investigated the effect of Ahr on collageninduced arthritis (CIA). We found that CIA was significantly suppressed in Ahr-deficient (KO) mice, compared to that in wild-type (WT) mice. Although WT mice with CIA showed the marked cartilage destruction, that histopathologic features were milder in Ahr KO mice than in WT mice. To determine which types of immune cells are important for the inhibition of the CIA development by Ahr-deficiency, we employed the conditional knockout strategy. Ahr gene was specifically deleted in macrophages or T cells by crossing Ahr-flox/flox mice with LysM-cre mice or Lck-
25
doi:10.1016/j.cyto.2010.07.108
University Graduate School of Dentistry, Sendai, Japan, 2 Division of Orthodontics and Dentofacial Orthopedics, Department of Oral Health and Development Sciences, Tohoku University Graduate School of Dentistry, Sendai, Japan
PS1-37 Aryl hydorcarbon receptor mediates suppression of colon inflamation induced by dextran sulfate sodium Ichino Chinen, Akihiro Kimura, Taisuke Nakahama, Kazuya Masuda, Tadamitsu Kishimoto, Laboratory of Immune Regulation, Graduate school of Frontier Bioscienses, Osaka University, Suita, Osaka, Japan
Objectives: We reported that lipopolysaccharide (LPS) promoted and augmented Ni-allergy in mice, and we thereby established a reproducible murine Ni-allergy model termed ‘Ni(+LPS)-allergy’. Histamine, an inflammatory mediator, also modifies Th1/ Th2 responses. It is widely known that histamine is pooled in mast cells or basophils, but less well known that the histamine-forming enzyme, histidine decarboxylase (HDC), is an inducible enzyme and that IL-1 and microbial substances (including LPS) can induce HDC in various murine tissues, even in mast cell-deficient mice. We found that HDC was induced at the site of Ni(+LPS)-allergy, and that Ni(+LPS)-allergy was induced in mast cell-deficient mice and even in nude mice that lack T-cells, but very weak either in HDC-deficient or in IL-1-deficent mice. Here, we report the roles of IL-1 and histamine in Ni(+LPS)-allergy. Methods: A mixture of NiCl2 and LPS was injected intraperitoneally into mice. Ten days later, NiCl2 was challenged intradermally into ears, and ear thickness was measured. In cell-transfer experiments, spleen cells from sensitized or non-sensitized mice (HDC-KO, histamine receptor H1-KO, IL1(both a and b types)-KO mice, or control wild type mice) were injected intravenously into non-sensitized mice, and 24h later, NiCl2 was challenged. Results: The results obtained from cell-transfer experiments demonstrated that (i) the elicitation step involves stimulation of H1 receptors by histamine newly formed via the induction of HDC, and (ii) IL-1 is involved in both the sensitization and elicitation steps. Conclusions: In murine Ni(+LPS)-allergy, IL-1 may be involved in the sensitization step through a histamine-independent mechanism, and it is likely that histamine, which is produced by an IL-1-dependent HDC induction, may be involved in the elicitation step.
Inflammatory bowel disease (IBD) is a chronic disorder of the gastrointestinal tract. It has become clear that Th1 or Th2, cause worsening symptoms, and moreover that IBDs are characterized by a sustained production of cytokines produced by newly identified Th17 cells. Our group and others have demonstrated that Aryl hydrocarbon receptor (Ahr) participates in Th17 cell differentiation and modulates the Th1/Th2 balance. In this study, we investigated the role of Ahr in the development of dextran sulfate sodium (DSS)-induced colitis. Wild type (WT) and Ahr deficient (KO) mice were given 1% DSS dissolved in drinking water. All Ahr KO mice, but not WT littermates, died within 9 days after DSS administration. Moreover, significant body weight loss in Ahr KO mice was observed following 4 days of DSS administration. Conversely, WT mice receiving 1% DSS showed no loss in body weight. Flow cytometry was used to determine the differentiation of T cells in colonic lamina propria lymphocytes (LPLs) from 1% DSS-treated WT and Ahr KO mice for 5 days. A significant increase in the percentage of Th1 cells was observed in DSS-treated Ahr KO mice compared with DSS-treated WT mice. However, there was no difference in Treg and Th17 cell development in LPLs from DSS-treated WT and Ahr KO mice. Furthermore, we found that pro-inflammatory cytokines including IL-12 were increased, in colon-cultured supernatant of 1%DSS treated Ahr KO mice compared to that of WT mice. It has also been reported that Ahr regulates the production of LPSinduced pro-inflammatory cytokines including IL-12, which induces Th1 cell development. Ahr may inhibit the development of Th1 cells through regulating IL-12 secretion from activated macrophages, which may result in DSS-induced colonic inflammation. We intend to further investigate the mechanistic detail of Ahr function in DSS-induced colitis using macrophage- and T cell- specific Ahr deficient mice.
doi:10.1016/j.cyto.2010.07.111
PS1-40 Stimulation of LY-6G on neutrophils in lipopolysaccharide-primed mice induces platelet-activating factor-mediated anaphylaxis-like shock Yukinori Tanaka, Yasuhiro Nagai, Toshinobu Kuroishi, Yasuo Endo, Shunji Sugawara, Division of Oral Immunology, Department of Oral Biology, Tohoku University Graduate School of Dentistry, Sendai, Japan
doi:10.1016/j.cyto.2010.07.109
PS1-38 Interleukin-6 blockade in CD4 T cells inhibits the development of experimental autoimmune uveitis Hiroshi Haruta 1, Nobuyuki Ohguro 1, Satoshi Hohki 1, Kohji Nishida 1, Minoru Fujimoto 2, Tetsuji Naka 2, 1 Department of Ophthalmology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan, 2 Laboratory for Immune Signal, National Institute of Biomedical Innovation, Ibaraki, Osaka, Japan Purpose: A novel subset of interleukin (IL)-17-producing CD4+ T helper cells (Th17) is thought to play a pivotal role in experimental autoimmune uveitis (EAU) and human uveitis. Recent studies also have indicated that IL-6 promotes Th17 differentiation in vitro and in vivo. The aim of this study is to investigate the role of IL-6 signaling in CD4 T cells in the development of EAU. Methods: EAU was induced by immunization with interphotoreceptor retinoid binding protein (IRBP) in wild type (WT), IL-6 knockout (KO) mice and T cell-specific signal transducer activated signal transducers and activator of transcription 3 (STAT3) KO female mice, which are defective in IL-6 signaling. All experimental mice used were on the C57BL/6 background. The clinical scores of EAU and CD4 T cell differentiation in these mice were analyzed. All mice were treated according to the ARVO Statement on the Use of Animals in Ophthalmic and Vision Research. Results: EAU did not develop in IL-6KO and STAT3KO mice while WT mice displayed severe inflammation. Th17 cell differentiation in cervical lymph nodes on day 10 after immunization also decreased in IL-6KO and STAT3KO compared to WT mice. IRBP specific CD4+CD25+ T regulatory cells were observed to be significantly increased in IL-6KO mice. Conclusion: This study showed that IL-6 and STAT3 in T cells are essential in the development of EAU and for the differentiation of highly pro-inflammatory Th17 cells. The protective effect of defective IL-6 signaling in the development of EAU is associated with the inhibition of differentiation of highly pro-inflammatory Th17 cells and an increase in the frequency of immunosuppressive T regulatory cells. doi:10.1016/j.cyto.2010.07.110
PS1-39 Roles of IL-1 and histamine in nickel (Ni) allergy in mice Masayuki Kinbara 1,2, Yasuhiro Nagai 1, Teruko Takano-Yamamoto 2, Yasuo Endo 1, Shunji Sugawara 1, 1 Division of Oral Immunology, Department of Oral Biology, Tohoku
Two anti-Ly-6G mAbs, RB6-8C5 and 1A8, have been used to deplete neutrophils in mice and to clarify their involvement in immune responses. During the course of experiments on neutrophil depletion, we noticed that intravenous injection of RB68C5 induced anaphylaxis-like shock in mice pretreated with lipopolysaccharide (LPS). We investigated the mechanism and obtained the following results. Signs of shock, such as hypothermia, appeared within a few minutes, and the mice died of shock within 20 min of the RB6-8C5 injection. Similar results were obtained with another anti-Ly-6G mAb, 1A8. Optimal LPS priming required at least 1 h. The priming effects of LPS may be associated with neutrophil accumulation in lung and liver (which began at 1 and 1.5 h after LPS injection, respectively). These neutrophils were localized within pulmonary and hepatic blood vessels. A platelet-activating factor (PAF)-receptor antagonist, CV-3988, prevented the shock. Moreover, a PAF biosynthetic enzyme, lyso-PAF acetyltransferase, was induced in the lungs by RB6-8C5 injection into mice pretreated with LPS. These results suggest that antiLy-6G mAbs may lead to PAF production by LPS-primed neutrophils located within pulmonary and hepatic blood vessels, resulting in rapid, lethal anaphylaxis-like shock. doi:10.1016/j.cyto.2010.07.112
PS1-41 Aryl hydrocarbon receptor deficiency significantly suppresses collagen induced arthritis in mice Nakahama T, Kimura A, Chinen I, Masuda K, Nguyen N, Kishimoto T, Laboratory of Immune Regulation, Graduate school of Frontier Bioscienses, Osaka University, Suita, Osaka, Japan Aryl hydrocarbon receptor (Ahr), as known a dioxin receptor, is a ligand-dependent transcription factor. It has been reported that Ahr plays an important role in the immune system. Our recent study demonstrated that Ahr participates in Th17 cell development and regulates inflammatory responses by lipopolysaccharide (LPS) in macrophage. In this study, we investigated the effect of Ahr on collageninduced arthritis (CIA). We found that CIA was significantly suppressed in Ahr-deficient (KO) mice, compared to that in wild-type (WT) mice. Although WT mice with CIA showed the marked cartilage destruction, that histopathologic features were milder in Ahr KO mice than in WT mice. To determine which types of immune cells are important for the inhibition of the CIA development by Ahr-deficiency, we employed the conditional knockout strategy. Ahr gene was specifically deleted in macrophages or T cells by crossing Ahr-flox/flox mice with LysM-cre mice or Lck-