FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2803043670> ?p ?o ?g. }

• W2803043670 abstract "Article19 April 2018free access Source DataTransparent process RKIP mediates autoimmune inflammation by positively regulating IL-17R signaling Wenlong Lin Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Department of Respiratory Medicine, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Ning Wang Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Kangxing Zhou Department of Rheumatology and Immunology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, China Search for more papers by this author Fasheng Su Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Yu Jiang Department of Clinical Laboratory Medicine, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Jianan Shou Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Huan Liu Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Chunmei Ma Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Youchun Qian The Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences/Shanghai Jiao Tong University School of Medicine, Shanghai, China Search for more papers by this author Kai Wang Department of Respiratory Medicine, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Xiaojian Wang Corresponding Author [email protected] orcid.org/0000-0001-8232-0558 Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Wenlong Lin Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Department of Respiratory Medicine, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Ning Wang Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Kangxing Zhou Department of Rheumatology and Immunology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, China Search for more papers by this author Fasheng Su Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Yu Jiang Department of Clinical Laboratory Medicine, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Jianan Shou Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Huan Liu Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Chunmei Ma Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Youchun Qian The Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences/Shanghai Jiao Tong University School of Medicine, Shanghai, China Search for more papers by this author Kai Wang Department of Respiratory Medicine, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Xiaojian Wang Corresponding Author [email protected] orcid.org/0000-0001-8232-0558 Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China Search for more papers by this author Author Information Wenlong Lin1,2,‡, Ning Wang1,‡, Kangxing Zhou3,‡, Fasheng Su1, Yu Jiang4, Jianan Shou1, Huan Liu1, Chunmei Ma1, Youchun Qian5, Kai Wang2 and Xiaojian Wang *,1 1Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China 2Department of Respiratory Medicine, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China 3Department of Rheumatology and Immunology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, China 4Department of Clinical Laboratory Medicine, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China 5The Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences/Shanghai Jiao Tong University School of Medicine, Shanghai, China ‡These authors contributed equally to this work *Corresponding author. Tel: +86 571 88206268; Fax: +86 571 88208285; E-mail: [email protected] EMBO Rep (2018)19:e44951https://doi.org/10.15252/embr.201744951 Correction(s) for this article RKIP mediates autoimmune inflammation by positively regulating IL-17R signaling05 February 2019 PDFDownload PDF of article text and main figures. Peer ReviewDownload a summary of the editorial decision process including editorial decision letters, reviewer comments and author responses to feedback. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract Th17 cells contribute to the development of autoimmune diseases by secreting interleukin-17 (IL-17), which activates its receptor (IL-17R) that is expressed on epithelial cells, macrophages, microglia, and resident neuroectodermal cells. However, the mechanisms through which IL-17R-mediated signaling contributes to the development of autoimmune disease have not been completely elucidated. Here, we demonstrate that Raf-1 kinase inhibitor protein (RKIP) deficiency in mice ameliorates the symptoms of experimental autoimmune encephalomyelitis (EAE). Adoptive T-cell-transfer experiments demonstrate that RKIP plays a predominant role in Th17-mediated, but not in Th1-mediated immune responses. RKIP deficiency has no effect on Th17-cell differentiation ex vivo, nor does it affect Th17-cell differentiation in EAE mice. However, RKIP significantly promotes IL-17R-induced proinflammatory cytokine and chemokine production. Mechanistically, RKIP directly interacts with IL-17RA and Act1 to promote the formation of an IL-17R-Act1 complex, resulting in enhanced MAPK- and P65-mediated NF-κB activation and downstream cytokine production. Together, these findings indicate that RKIP functions as an essential modulator of the IL-17R-Act1 axis in IL-17R signaling, which promotes IL-17-induced inflammation and autoimmune neuroinflammation. Synopsis The Raf-1 kinase inhibitor protein participates in the pathogenesis of IL-17-mediated autoimmune diseases and inflammation by promoting the formation of the IL-17RA-Act1 complex, required for downstream signaling and cytokine production. RIPK promotes EAE pathogenesis via enhanced IL-17R-mediated signaling and inflammation. RKIP positively regulates IL-17-induced inflammation in vitro and in vivo. RKIP interacts with IL-17R and Act1, thereby stabilizing the IL-17R-Act1 complex. Introduction Multiple sclerosis (MS) is a type of human demyelinating disease that causes central nervous system (CNS; i.e., the brain and spinal cord) inflammation, neuronal demyelination, and axonal damage, resulting in both physical and mental health impairment. MS is the most common autoimmune disorder affecting the central nervous system 1. Experimental autoimmune encephalomyelitis (EAE) is a well-established MS-like animal disease model induced by immunizing mice with myelin oligodendrocyte glycoprotein peptide (MOG35-55) and pertussis toxin (PTX) 2. It has been demonstrated that myelin-specific T cells, particularly interleukin-17 (IL-17)-producing Th17 cells, play a crucial pathogenic role in the mouse EAE model 3, 4. Specifically, Th17 cells participate in the development of EAE by secreting IL-17, which comprises IL-17A to IL-17F 5, 6. IL-17 levels have been shown to be increased in patients with MS and ulcerative colitis (UC) 7, 8. The binding of IL-17 to a type I cell surface receptor known as the IL-17 receptor (IL-17R) activates IL-17R signaling and ultimately triggers the production of proinflammatory cytokines (TNF-α, IL-1, IL-6, IL-8, G-CSF, and GM-CSF) and chemokines (CXCL1, CXCL2, CXCL5, CCL2, CCL7, and CCL20) 9. These cytokines subsequently contribute to the activation of resident CNS cells, including epithelial cells, macrophages, microglia, and astrocytes, resulting in the secretion of additional chemokines and the recruitment of additional leukocytes into the CNS, which leads to disseminated CNS inflammation, neuronal demyelination, and the development of EAE symptoms 3, 6, 10. Consistent with these findings, the findings of additional studies indicate that mice deficient in IL-17 or IL-17R display less severe EAE than wild-type (WT) mice because they lack the cellular machinery responsible for facilitating IL-17-mediated IL-17R signal transduction 11, 12. IL-17R signaling involves the recruitment of the key adaptor protein Act1 (also known as CIKS) and the E3 ubiquitin ligase TRAF6 to the cytoplasmic region of IL-17R, where they bind with its SEFIR domain. Specifically, the SEFIR domain of IL-17R interacts with the adaptor protein Act1 in response to IL-17 stimulation 13, 14. Act1 subsequently recruits TRAF6 to IL-17R, then triggers activation of the MAPK and NF-κB pathways, and induces the production of proinflammatory cytokines and chemokines 15, 16. In addition to activating TRAF6, Act1 also recruits TRAF2 and TRAF5 to prolong the half-life of the mRNA of some cytokines and chemokines, such as IL-6 and CXCL1 17, 18. Previous studies have demonstrated the regulatory mechanisms through which multiple signaling molecules downstream of the IL-17R-Act1 complex exert their effects. For example, Xiao et al 10 reported that the serine/threonine kinase TPL2 functions as a pivotal mediator of TAK1 activation in the IL-17R signaling pathway. However, the mechanism through which the IL-17R-Act1 signalosome is modulated remains poorly understood 14. Raf-1 kinase inhibitor protein (RKIP) is a member of the phosphatidylethanolamine-binding protein (PEBP) family and is widely expressed in both prokaryotic and eukaryotic organisms. RKIP modulates and controls crucial intracellular signaling networks, including the Raf/MEK/ERK, NF-κB, glycogen synthase kinase-3β (GSK3β), and G protein-coupled receptor (GPCR) signaling cascades 19-22. Therefore, RKIP is involved in regulating a variety of physiological processes, such as cell differentiation, cell cycle progression, and apoptosis 23. RKIP has been shown to inhibit cancer cell invasion and metastasis in several cancers and functions as an important tumor suppressor 24, 25. We recently determined that RKIP also plays a crucial role in mediating human and mouse colitis by promoting intestinal epithelial cell apoptosis 26. Furthermore, we determined that RKIP promotes anti-viral innate immune responses by promoting TBK1 auto-phosphorylation 27. Therefore, RKIP functions in more processes than tumor suppression. In the present study, we first obtained genetic evidence that RKIP deficiency in non-hematopoietic cells renders affected mice refractory to EAE induction. Adoptive T-cell-transfer experiments demonstrated that RKIP promotes Th17-mediated EAE development. We further demonstrated that RKIP deficiency in astrocytes significantly inhibits IL-17-induced proinflammatory cytokine and chemokine expression. More importantly, we found that RKIP serves as a pivotal mediator of the interactions between IL-17R-Act1 in the IL-17R signaling pathway, resulting in enhanced IL-17-induced NF-κB and the MAPK activation. RKIP deficiency in mice significantly impairs IL-17-induced pulmonary inflammation and peritonitis. IL-17R signaling is involved in various inflammatory diseases, and these findings not only provided novel insights into the regulatory mechanisms through which IL-17R signaling occurs, but also showed that targeting IL-17R signaling may be useful for the treatment of IL-17-mediated autoimmune diseases. Results RKIP is a crucial mediator of the pathogenesis of EAE We previously determined that RKIP is involved in colitis and anti-viral immune responses 26, 27. We thus attempted to determine whether RKIP can regulate autoimmune diseases. The well-known mouse MS model, that is, the EAE model, was firstly established to determine the role of RKIP in the pathogenesis of EAE. We subcutaneously injected RKIP WT mice and their knockout (KO) littermates with MOG35-55 and then intravenously injected them with pertussis toxin (PTX) to induce the EAE model. As shown in Fig 1A and B, RKIP-KO mice suffered from relatively mild EAE disease symptoms compared to WT mice and had lower EAE clinical scores than their counterparts. Moreover, WT mice displayed diminished inflammatory cell infiltration (Fig 1C) and demyelination (Fig 1D), as demonstrated by hematoxylin–eosin (H&E) and luxol fast blue (LFB) staining, respectively, compared to their counterparts. Consistent with these results, flow cytometry analysis of mouse CNS tissues (brains and spinal cords) demonstrated that both the percentages and the absolute numbers of CNS-infiltrating CD11b+Gr-1+ neutrophils were significantly reduced in RKIP-KO mice compared to WT mice (Fig 1E). Within the CNS-infiltrating CD4+ T-cell population, the percentages and absolute numbers of IL-17+ Th17 cells and Foxp3+ Treg cells in RKIP-KO mice were comparable to those in WT mice (Fig 1F), while the numbers of Th1 (IFNγ+ CD4+ T) cells in RKIP-KO mice were slightly decreased compared to those in WT mice (Fig 1F). We also assessed the composition of the immune cell population in the spleen during EAE. The results of the assessment showed that RKIP deficiency inhibited CD11b+Gr-1+ neutrophil infiltration into the spleen (Fig EV1A–C) without affecting the absolute numbers or percentages of other immune cells, namely CD4+ T cells, CD8+ T cells, and CD11b+Gr-1− monocytes in the spleen. RKIP deficiency either did not alter T-helper-cell differentiation (namely Th1-, Th17-, and Treg-cell differentiation; Fig EV1D and E). We subsequently examined the expression levels of several proinflammatory cytokines and chemokines in the CNS tissues of WT and RKIP-KO EAE mice. As shown in Fig 1G, the gene expression levels of the proinflammatory cytokine IL-6 and several chemokines known to mediate immune cell recruitment, such as CXCL1 (KC) and CXCL2, were significantly decreased in RKIP-KO EAE mice versus WT EAE mice. These results suggest that RKIP deficiency restricts EAE development by reducing the gene expression levels of several proinflammatory cytokines and chemokines. Figure 1. RKIP is a crucial mediator of the pathogenesis of EAE A. The mean clinical scores for RKIP-KO (KO, n = 5) mice and their wild-type littermates (WT, n = 7) induced by being immunized with MOG35-55 were assessed from day 0 to day 31 after immunization. B. Linear regression curves of (A) dashed lines indicate the 95% confidence intervals of the regression lines. C, D. Histology of the spinal cord from WT and KO mice on day 31 after MOG35-55 immunization was analyzed by hematoxylin–eosin (H&E) (C) and luxol fast blue (LFB) (D) staining. Scale bars (a whole spinal cord, 100X), 100 μm; scale bars (a portion of the spinal cord, 200X), 50 μm. E. In a separated experiment, summary graph of the percentages of cells (left) and the absolute numbers of cells (right) in the CNS (brains and spinal cords). CNS-infiltrating cells isolated from mice treated as in (A) on day 31 were stained with mouse anti-CD4, anti-CD8, anti-CD11b, and anti-Gr-1 antibodies and analyzed by flow cytometry (FACS; WT n = 5, KO n = 5). F. T-helper cells isolated from the CNS of mice treated as in (A) on day 31 were fixed and permeabilized, and the CD4+ T cells were analyzed by flow cytometry to measure intracellular IFN-γ, IL-17, and Foxp3 expression. The data are presented in summary graphs of percentages (left) and absolute cell numbers (right; WT n = 5, KO n = 5). G. The expression levels of proinflammatory cytokines and chemokines in the CNS tissues of WT and KO EAE mice, as determined by real-time PCR (WT n = 5, KO n = 5). Data information: *P < 0.05, **P < 0.01, ***P < 0.001; ns, no significant difference (unpaired, two-tailed Student's t-test). Data are representative of three independent experiments with similar results. Data are means ± SEM values. Download figure Download PowerPoint Click here to expand this figure. Figure EV1. RKIP deficiency has no effect on T cell development, polarization, and immune cell composition in the spleen from EAE mice A–C. Immune cells isolated from spleen of day 31 MOG35-55-immunized WT and RKIP-KO mice were stained with mouse anti-CD4, anti-CD8, anti-CD11b, and anti-Gr-1 antibodies and analyzed by flow cytometry (FACS; WT n = 6, KO n = 5). Data are presented as the representative plot (A), summary graph of the percentages of cells (B), and the absolute numbers of cells (C). D, E. T-helper cells isolated from spleen of day 31 MOG35-55-immunized WT and RKIP-KO mice were fixed and permeabilized, and the CD4+ T cells were analyzed by flow cytometry to measure intracellular IFN-γ, IL-17, and Foxp3. Data are presented in summary graphs of percentages (D) and absolute cell numbers (E) (WT n = 6, KO n = 5). F, G. T cells from thymus, spleen, and dLNs of WT and RKIP-KO mice (n = 5/per group) were analyzed with anti-CD4 and anti-CD8 staining. H, I. The naïve CD4+ T cells were polarized under the Th1, Th2, Th17, or Treg condition, respectively, and 5 days later, the CD4+ T cells were fixed and permeabilized, followed by flow cytometry to measure intracellular IFN-γ, IL-4, IL-17, and Foxp3. Data are presented as the representative plot (H) and summary graph of the percentages of cells (I), Th1 (n = 4), Th2 (n = 2), Th17 (n = 4), and Treg (n = 2). Data information: *P < 0.05; ns, no significant difference (unpaired, two-tailed Student's t-test). Data are representative of three independent experiments with similar results. Data are means ± SEM values. Download figure Download PowerPoint RKIP deficiency in non-hematopoietic cells impairs the pathogenesis of EAE We subsequently attempted to identify the cellular compartment in which RKIP meditates the pathogenesis of EAE. We generated bone marrow chimeric mice by transferring WT or RKIP-KO bone marrow cells to lethal dose-irradiated (9.5 Glyc) WT or RKIP-KO recipient mice. After 8 weeks of reconstitution, the chimeric mice were immunized with MOG35-55 and injected with PTX to induce EAE. WT mice reconstituted with WT bone marrow cells (WT→WT group) or RKIP-KO bone marrow cells (KO→WT group) displayed comparable EAE clinical scores, while RKIP-KO mice reconstituted with WT bone marrow cells (WT→KO group) displayed lower EAE clinical scores than their counterparts (Fig 2A and B). H&E and LFB staining showed that inflammatory cell infiltration and neuronal demyelination were ameliorated in the WT→KO group compared to the KO→WT and WT→WT groups (Fig 2C and D). Consistently, the percentage of CNS-infiltrating CD11b+Gr-1+ neutrophils was significantly reduced in the WT→KO group compared to that in the KO→WT and WT→WT groups (Fig 2E and F). However, neuronal demyelination and infiltration of inflammatory cells were comparable between the spinal cords of KO→WT chimeras and that of WT→WT (Fig 2C and D). Moreover, the percentages and absolute numbers of IL-17+ Th17 cells, IFN-γ+ Th1 cells, and Foxp3+ Treg cells in the CNS-infiltrating CD4+ T-cell population were comparable among the three groups (Fig 2G). These results indicate that RKIP in non-hematopoietic cells regulates the development of EAE. Figure 2. RKIP deficiency in non-hematopoietic cells mediates the pathogenesis of EAE A. Mice with reconstituted bone marrow cells (WT→WT n = 7, WT→KO n = 7, and KO→WT n = 9) were immunized with MOG-35-55 to induce EAE. The mean clinical scores were calculated every other day. B. Linear regression curves of (A) dashed lines indicate the 95% confidence intervals of the regression lines. C, D. Histology of spinal cord sections from MOG35-55-immunized mice was analyzed by H&E (C) and LFB (D) staining. Scale bars (a whole spinal cord, 100X), 200 μm; scale bars (a portion of the spinal cord, 400X), 50 μm. E. Gr-1 immunofluorescence staining of spinal cord sections from MOG35-55-immunized mice treated as in (A). The nuclei were counterstained with DAPI (blue). Scale bars, 100 μm. F. In a separated experiment, CNS cells isolated from chimeric mice on day 18 after MOG35-55-immunization were analyzed as in Fig 1D (WT n = 5, KO n = 5). G. CNS T-helper cells isolated from chimeric mice on day 18 after MOG35-55 immunization were analyzed as in Fig 1E (WT n = 5, KO n = 5). Data information: Data are representative of two independent experiments with similar results. Data are means ± SEM values. Download figure Download PowerPoint Homozygous RKIP-KO mice were viable and did not display any abnormalities with respect to growth or survival, suggesting that RKIP is not essential for mouse growth and development. There are no underlying deficits in T-cell, B-cell, NK-cell, or CD11b+ APC populations in homozygous RKIP-KO mice 28. The compositions of the T-cell populations in various organs (including the thymus, spleen, and lymph nodes) in 2- to 3-month-old RKIP-KO mice were similar to those in their WT littermates (Fig EV1F and G), indicating that RKIP deficiency in mice does not affect the development of various types of immune cells. CD44loCD62Lhi naive CD4+ T cells were sorted by FACS, and in vitro polarization assays were performed under different T-cell polarization conditions, including Th1, Th2, Th17, and Treg polarization conditions. As shown in Fig EV1H and I, knocking out RKIP had no significant effect on Th1-, Th2-, Th17-, and Treg-cell production. Taken together, these results indicate that RKIP deficiency does not affect T-cell activation and differentiation in vivo and in vitro. RKIP positively regulates the pathogenesis of EAE via IL-17R-mediated inflammation To elucidate the mechanisms through which RKIP contributes to EAE development, we transferred pre-activated MOG35-55-specific Th17 cells or Th1 cells into γ-irradiated (5Gylc) WT or RKIP-KO recipient mice to induce EAE. After 6–8 days of MOG35-55-specific Th17-cell transfer, both WT and RKIP-KO recipient mice (Th17→WT and Th17→KO) exhibited severe EAE. However, RKIP-KO recipient mice developed relatively mild EAE compared to WT recipient mice and exhibited lower clinical scores than their counterparts (Fig 3A and B). Moreover, CNS-infiltrating immune cell recruitment, particularly CD11b+Gr-1− monocytes, and CD11b+Gr-1+ neutrophils recruitment, was significantly decreased in RKIP-KO recipient mice compared to WT recipient mice (Fig 3C and D). Diminished inflammation and demyelination were also observed in RKIP-KO recipient mice compared to WT recipient mice (Fig 3E). The gene expression levels of chemokines, such as CXCL2 and CCL20, in the CNS tissues of Th17→KO EAE mice were significantly lower than those in the CNS tissues of Th17→WT EAE mice (Fig 3F). We also transferred WT MOG35-55-specific Th1 cells into γ-irradiated (5Gylc) WT or RKIP-KO recipient mice and then assessed the EAE clinical scores of these mice. As shown in Fig EV2A and B, EAE clinical scores were comparable between the two groups (Th1→WT and Th1→KO). Recruitment of CD4+ T cells, CD8+ T cells, CD11b+Gr-1− monocytes, and CD11b+Gr-1+ neutrophils was also comparable between RKIP-KO recipient mice and WT recipient mice (Fig EV2C and D). Moreover, inflammation, demyelination, and chemokine gene expression levels in the CNS tissues (Th1→WT and Th1→KO) were almost identical between the two groups (Fig EV2E and F). Taken together, these results indicate that RKIP plays a predominant role in the Th17-mediated immune response but not in the Th1-mediated immune response. Figure 3. RKIP deficiency inhibits Th17 cell passive transfer-induced EAE A. The mean EAE clinical scores for γ-irradiated (5Gylc) WT and RKIP-KO mice (WT n = 8, KO n = 7) reconstituted with WT MOG-35-55-pre-sensitized Th17 cells were determined from day 0 to day 18 after Th17-cell transfer. B. Linear regression curves of (A) dashed lines indicate the 95% confidence intervals of the regression lines. C, D. Summary graph of the percentages of cells (C) and absolute numbers of cells (D) in the CNS. CNS-infiltrating cells isolated from mice treated as in (A) (WT n = 8, KO n = 7) were stained with the appropriate antibodies. E. Histology of the spinal cord from mice reconstituted with Th17 cells was analyzed by H&E (left) and LFB (right) staining. Scale bars (a whole spinal cord, 100X), 200 μm; scale bars (a portion of the spinal cord, 400X), 50 μm. F. Real-time PCR analysis of CXCL2 and CCL20 mRNA expression in the CNS tissues of WT and RKIP-KO mice treated as in (A) (WT n = 8, KO n = 7). G. The mice (WT n = 5, KO n = 4) were treated with intraperitoneal (i.p.) injection of an anti-IL-17A antibody (100 μg per mouse each time) or appropriate isotype controls on days 7, 9, 11, and 13 after the second MOG-35-55 immunization. Mean clinical scores were calculated every other day according to the standards described in the Materials and Methods section. H. Histology of the spinal cord was analyzed by H&E (left) or LFB (right) staining on day 14 after the second MOG-35-55 immunization. Scale (a whole spinal cord, 100X), 100 μm; scale (a portion of the spinal cord, 200X), 50 μm. I. IL-6, CXCL2, and TNF-α mRNA in the CNS tissues (including brain and spinal cords) were measured by real-time PCR on day 14 after the second MOG-35-55 immunization, (WT n = 5, KO n = 4). Data information: Data are representative of two independent experiments with similar results. Data are means ± SEM values. Download figure Download PowerPoint Click here to expand this figure. Figure EV2. RKIP deficiency does not affect Th1 cells passive transferring-induced EAE A. The mean EAE clinical scores of γ-irradiated (5Gylc) WT and RKIP-KO mice (WT n = 3, KO n = 4) reconstituted with WT MOG-35-55-pre-sensitized Th1 cells were determined from day 0 to day 18 after Th1-cell transfer. B. Linear regression curves of (A) dashed lines indicate the 95% confidence intervals of the regression lines. C, D. Summary graph of the percentages of cells (C) and the absolute numbers of cells (D) in the CNS. CNS-infiltrating cells isolated from mice treated as in (A) on day 18 after the Th1-cell transfer was stained with the appropriate antibody. E, F. Histology of the spinal cord from mice reconstituted with Th1 cells was analyzed by H&E (E) and LFB (F) staining of spinal cord sections from mice reconstituted with Th1 cells. Scale bars (a whole spinal cord, 100X), 200 μm; scale bars (a portion of the spinal cord, 400X), 50 μm. Typical stainings in the black box region were magnified and shown in 400X panel. G. Real-time PCR analysis of CXCL2 and CCL20 mRNA expression in the CNS tissues of WT and RKIP-KO mice treated as in (A). Data information: Data are representative of two independent experiments with similar results. Download figure Download PowerPoint Th17 cells secret IL-17A which initiates the IL-17R signaling and plays an important role in the EAE pathogenesis 6. We next investigated whether the reduced EAE pathogenesis in RKIP deficiency mice was dependent on IL-17R-mediated signaling and its inflammation, an anti-α-IL-17A-specific blocking antibody was continually i.p. injected into RKIP WT and KO mice during the induction of EAE. As shown in Fig 3G and H, the injection of anti-α-IL-17A antibody resulted in a significant inhibited EAE symptom, including clinical scores, inflammation, and demyelination, in RKIP WT and KO mice compared to the anti-isotype control antibody-injected RKIP WT group. The RKIP-deficient mice exhibited much reduced EAE severity compared to WT mice, which was obliterated after the injection of IL-17A-blocking antibody. Consistently, the induction of IL-17A-induced genes, such as IL-6, CXCL2, and TNF-α, in CNS tissues was substantially attenuated in RKIP-KO mice compared to WT mice. And the expression of these genes in WT mice decreased to comparable levels in KO mice after treatment with IL-17A-blocking antibody (Fig 3I). Collectively, these data suggest RKIP regulates EAE pathogenesis via IL-17R-mediated signaling-associated inflammation. RKIP positively regulates IL-17-induced gene expression and protein production Myelin-specific Th17 cells secrete IL-17A, which bind IL-17R-expressing epithelial cells, macrophages, microglia, and resident neuroectodermal cells (neurons, astrocytes, and oligodendrocytes) and induce proinflammatory cytokine and chemokine gene expression in these resident CNS cells, particularly in astrocytes, a well-known cell type that has been reported to be crucial for IL-17-induced EAE 10, 29. Given that RKIP regulates the pathogenesis of EAE via IL-17R signaling, we assessed the involvement of RKIP in IL-17-induced inflammation in astrocytes by stimulating primary astrocytes isolated from WT and RK" @default.
• W2803043670 created "2018-05-17" @default.
• W2803043670 creator A5006150870 @default.
• W2803043670 creator A5011564728 @default.
• W2803043670 creator A5012645572 @default.
• W2803043670 creator A5013881064 @default.
• W2803043670 creator A5019257366 @default.
• W2803043670 creator A5023713311 @default.
• W2803043670 creator A5024159517 @default.
• W2803043670 creator A5043839950 @default.
• W2803043670 creator A5057653584 @default.
• W2803043670 creator A5063203754 @default.
• W2803043670 creator A5063949651 @default.
• W2803043670 date "2018-04-19" @default.
• W2803043670 modified "2023-10-18" @default.
• W2803043670 title "<scp>RKIP</scp> mediates autoimmune inflammation by positively regulating <scp>IL</scp> ‐17R signaling" @default.
• W2803043670 cites W1848518902 @default.
• W2803043670 cites W1937254170 @default.
• W2803043670 cites W1959664751 @default.
• W2803043670 cites W1975204978 @default.
• W2803043670 cites W1979652500 @default.
• W2803043670 cites W1982205351 @default.
• W2803043670 cites W1993662841 @default.
• W2803043670 cites W1998000193 @default.
• W2803043670 cites W1999845762 @default.
• W2803043670 cites W2021674244 @default.
• W2803043670 cites W2024595613 @default.
• W2803043670 cites W2026778930 @default.
• W2803043670 cites W2028215082 @default.
• W2803043670 cites W2034619319 @default.
• W2803043670 cites W2040332267 @default.
• W2803043670 cites W2058487327 @default.
• W2803043670 cites W2060500374 @default.
• W2803043670 cites W2060527543 @default.
• W2803043670 cites W2065136791 @default.
• W2803043670 cites W2081351081 @default.
• W2803043670 cites W2083361647 @default.
• W2803043670 cites W2093114816 @default.
• W2803043670 cites W2096618637 @default.
• W2803043670 cites W2096629377 @default.
• W2803043670 cites W2103961090 @default.
• W2803043670 cites W2105663540 @default.
• W2803043670 cites W2117895944 @default.
• W2803043670 cites W2119763611 @default.
• W2803043670 cites W2136935687 @default.
• W2803043670 cites W2142242441 @default.
• W2803043670 cites W2145468317 @default.
• W2803043670 cites W2148289263 @default.
• W2803043670 cites W2159598329 @default.
• W2803043670 cites W2160247072 @default.
• W2803043670 cites W2164363164 @default.
• W2803043670 cites W2169477525 @default.
• W2803043670 cites W2193056702 @default.
• W2803043670 cites W2310017168 @default.
• W2803043670 cites W2532725874 @default.
• W2803043670 cites W2589236405 @default.
• W2803043670 cites W4211254511 @default.
• W2803043670 doi "https://doi.org/10.15252/embr.201744951" @default.
• W2803043670 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6362353" @default.
• W2803043670 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/30723107" @default.
• W2803043670 hasPublicationYear "2018" @default.
• W2803043670 type Work @default.
• W2803043670 sameAs 2803043670 @default.
• W2803043670 citedByCount "14" @default.
• W2803043670 countsByYear W28030436702018 @default.
• W2803043670 countsByYear W28030436702019 @default.
• W2803043670 countsByYear W28030436702020 @default.
• W2803043670 countsByYear W28030436702021 @default.
• W2803043670 countsByYear W28030436702022 @default.
• W2803043670 countsByYear W28030436702023 @default.
• W2803043670 crossrefType "journal-article" @default.
• W2803043670 hasAuthorship W2803043670A5006150870 @default.
• W2803043670 hasAuthorship W2803043670A5011564728 @default.
• W2803043670 hasAuthorship W2803043670A5012645572 @default.
• W2803043670 hasAuthorship W2803043670A5013881064 @default.
• W2803043670 hasAuthorship W2803043670A5019257366 @default.
• W2803043670 hasAuthorship W2803043670A5023713311 @default.
• W2803043670 hasAuthorship W2803043670A5024159517 @default.
• W2803043670 hasAuthorship W2803043670A5043839950 @default.
• W2803043670 hasAuthorship W2803043670A5057653584 @default.
• W2803043670 hasAuthorship W2803043670A5063203754 @default.
• W2803043670 hasAuthorship W2803043670A5063949651 @default.
• W2803043670 hasBestOaLocation W28030436701 @default.
• W2803043670 hasConcept C185592680 @default.
• W2803043670 hasConcept C203014093 @default.
• W2803043670 hasConcept C2776914184 @default.
• W2803043670 hasConcept C62478195 @default.
• W2803043670 hasConcept C86803240 @default.
• W2803043670 hasConcept C95444343 @default.
• W2803043670 hasConceptScore W2803043670C185592680 @default.
• W2803043670 hasConceptScore W2803043670C203014093 @default.
• W2803043670 hasConceptScore W2803043670C2776914184 @default.
• W2803043670 hasConceptScore W2803043670C62478195 @default.
• W2803043670 hasConceptScore W2803043670C86803240 @default.
• W2803043670 hasConceptScore W2803043670C95444343 @default.
• W2803043670 hasFunder F4320321001 @default.
• W2803043670 hasFunder F4320321540 @default.
• W2803043670 hasFunder F4320323539 @default.
• W2803043670 hasFunder F4320338464 @default.
• W2803043670 hasIssue "6" @default.

• next