DOCSLIB.ORG

Interleukin-17A Induces IL-1B Secretion from RPE Cells Via the NLRP3 Inflammasome

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Interleukin-17A Induces IL-1B Secretion from RPE Cells Via the NLRP3 Inflammasome Retinal Cell Biology Interleukin-17A Induces IL-1b Secretion From RPE Cells Via the NLRP3 Inflammasome Shujie Zhang,1–3 Ning Yu,4 Rong Zhang,1–3 Shenghai Zhang,1–3 and Jihong Wu1–3 1Research Center, Eye & ENT Hospital of Fudan University, Shanghai, China 2Key Laboratory of Myopia, Ministry of Health, Fudan University, Shanghai, China 3Shanghai Key Laboratory of Visual Impairment and Restoration, Fudan University, Shanghai, China 4Department of Dermatology, Shanghai Skin Disease Hospital, Shanghai, China Correspondence: Jihong Wu, Re- PURPOSE. Inflammasome activation and IL-1b production have been proposed to have an search Center, Eye & ENT Hospital, important role in age-related macular degeneration (AMD). Growing evidence is emerging for Shanghai Medical College, Fudan involvement of interleukin-17A (IL-17A) in AMD pathogenesis. We investigated the effects of IL- University,83FenyangRoad,Shang- 17A on the activation of inflammasome and production of IL-1b in primary human RPE cells. hai 200031, China; [email protected]. METHODS. Primary human RPE cells were isolated and cultured for the following experiments. Shenghai Zhang, Research Center, Expression patterns of IL-17 receptor A (IL-17RA), IL-17 receptor C (IL-17RC), and ACT1 were Eye & ENT Hospital, Shanghai Med- analyzed by RT-PCR, flow cytometry, and immunofluorescence. IL-17A was added to the cell ical College, Fudan University, 83 cultures, and cytokine expression, signaling pathways, and inflammasome machinery were Fenyang Road, Shanghai 200031, investigated using real-time RT-PCR, ELISA, Western blot, flow cytometry, and small China; interfering RNA. [email protected]. RESULTS. Retinal pigment epithelial cells constitutively expressed IL-17RA, IL-17RC, and ACT1. SZ and NY contributed equally to the IL-17A upregulated the mRNA levels of pro-IL-1b, IL-8, CCL2, and CCL20, as well as the work presented here and should protein level of IL-1b. IL-17A induced the phosphorylation of Akt, Erk1/2, p38 MAPK, and NF- therefore be regarded as equivalent jB p65 in RPE cells. Blocking NF-jB attenuated IL-17A–induced expression of pro-IL-1b authors. mRNA. IL-17A enhanced pro-caspase-1 and NLRP3 mRNA expression. Inhibiting caspase-1 Submitted: June 27, 2015 activity and silencing NLRP3 decreased IL-1b secretion, confirming NLRP3 as the IL-17A– Accepted: December 8, 2015 responsive inflammasome on the posttranscriptional level. The mechanism of IL-17A– triggered NLRP3 activation and subsequent IL-1b secretion was found to involve the Citation: Zhang S, Yu N, Zhang R, Zhang S, Wu J. Interleukin-17A induc- generation of reactive oxygen species. es IL-1b secretion from RPE cells via CONCLUSIONS. Our results suggest that IL-17A triggers a key inflammatory mediator, IL-1b, from the NLRP3 inflammasome. Invest RPE cells, via NLRP3 inflammasome activation, holding therapeutic potential for AMD. Ophthalmol Vis Sci. 2016;57:312– 319. DOI:10.1167/iovs.15-17578 Keywords: RPE cells, IL-17A, inflammasome, AMD ge-related macular degeneration (AMD) is a neurodegener- it still is not clear whether this component of innate immunity A ative disease with a multifactorial etiology, primarily could be activated or regulated by the local inflammatory attributable to morphologic and functional abnormalities in cytokine milieu generated during adaptive immune responses. the RPE cells.1 Mounting evidence has highlighted the essential Interleukin-17A (IL-17A) is the signature cytokine of Th17 role of immune processes in the development, progression, and cells and has a pivotal role in inducing expression of treatment of AMD. Innate and adaptive immune systems have proinflammatory cytokines and chemokines in the pathogenesis been shown to contribute to AMD pathogenesis.2,3 of autoimmune and inflammatory diseases.16 Recent studies Inflammasome activation is a key component of innate have demonstrated involvement of IL-17A in the pathogenesis of immunity.4 Recent efforts have drawn attention to the AMD. Aberrant levels of IL-17A were observed in AMD macular inflammasome machinery, particularly its product IL-1b,asan lesions and sera.17 Complement component C5a, which inciting factor in AMD.5–8 It has been well documented that accumulated in AMD patient serum and in drusen, induced IL- RPE cells can be triggered to secrete a number of inflammatory 17A production from CD4þ T cells.18 Arecentepigeneticstudy cytokines and chemokines.9 Among these, IL-1b is one of the demonstrated that hypomethylation of the IL-17 receptor C (IL- initial cytokines produced following RPE activation.10,11 Tran- 17RC) promoter was associated with AMD.19 Since previous scription of the pro-IL-1b gene and production of cytosolic pro- reports showed the presence of IL-17A receptors on RPE IL-1b are dependent on the activation of NF-jB. At the cells,20,21 possibility exists that IL-17A may have a role in AMD posttranscriptional level, generation of mature IL-1b is regulat- by direct modulating the immunologic functions of RPE cells. ed by inflammasomes, which is formed by NACHT, LRR, and Both IL-17A signaling and inflammasome machinery have PYD domains-containing (NLRP) proteins, the most studied been linked to the pathogenesis of AMD; however, to our member of which is NLRP3.12 More recently, the role of the knowledge no studies to date have adequately addressed the NLRP3 inflammasome in AMD pathogenesis has been investi- interactions between them. Interleukin-1b functions in synergy gated extensively using AMD-related stimuli, such as A2E,8 Alu with IL-23 to promote the production of IL-17A and related RNA,13 lysosomal activation,14 and oxidative stress.15 However, cytokines from Th17 cells.22,23 Therefore, we hypothesized that iovs.arvojournals.org j ISSN: 1552-5783 312 This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. Downloaded from tvst.arvojournals.org on 10/01/2021 IL-17A Induces IL-1b Secretion From RPE Cells IOVS j February 2016 j Vol. 57 j No. 2 j 313 TABLE. The Sequence of Primers Used for RT-PCR and Real-Time RT- removed, the posterior poles were incubated in 2% dispase PCR solution (Gibco, Grand Island, NY, USA) for 30 minutes in 378C, 5% CO2. After dispase treatment, the posterior poles were Gene Primers dissected into quadrants, and the retina was removed gently with IL-17RA 50-CGT TTG TGC GTC AGG TTT G-30 forceps. Single-cell RPE layers were peeled off in small sheets, 50-GCA GGA TCT GGT AAT GGG TAG A-30 and then RPE cells were dissociated from the sheets in a solution IL-17RC 50-AGA TGC CTG TGC CCT GGT TCT TGC T-30 containing 0.25% trypsin and 0.02% ethylenediaminetetraacetic 50-AGC AAG AAC CAG GGC ACA GGC ATC T-30 acid (Life Technologies, Grand Island, NY, USA) at 378Cfor30 ACT1 50-CCG TGA TGA TAA TCG TAG C-30 minutes. Afterward, the enzyme-treated tissues were dissociated 50-TAG CAT TTG GGA AGA GCA-30 into single RPE cells by gentle pipetting and centrifugation. Pro-caspase-1 50-TGA AGG ACA AAC CGA AGG-30 The RPE cells were cultured in DMEM:F12 growth medium 50-GAA GAG CAG AAA GCG ATA-30 containing 5.5 mM glucose (Gibco), supplemented with 10% NLRP3 50-GCC TCA ACA AAC GCT ACA-30 fetal bovine serum (Gibco). The cell cultures were maintained 50-GAC GCC CAG TCC AAC ATC-30 in a humidified incubator at 378C in an atmosphere containing Pro-IL-1b 50-AGC TAC GAA TCT CCG ACC AC-30 95% air and 5% CO2. The cells were passed every 3 days by 50-CGT TAT CCC ATG TGT CGA AGA A-30 trypsinization. For the following experiments, RPE cells were IL-8 50-ACT CCA AAC CTT TCC ACC-30 seeded into 6-well plates (Corning, Lowell, MA, USA). The 50% 50-CTT CTC CAC AAC CCT CTG-30 confluent RPE cell cultures were used in RPE65 immunoflu- CCL2 50-CTT CTG TGC CTG CTG CTC-30 orescence and the 80% to 100% confluent RPE cell cultures at 50-TGC TGC TGG TGA TTC TTC T-30 passages 3 to 5 were used in other experiments. CCL20 50-TGC TGC TAC TCC ACC TCT-30 After treatment with IL-17A (up to 200 ng/mL), Bay 11-7082 50-GCA AGT GAA ACC TCC AAC-30 (up to 10 lM), Z-YVAD-fmk (up to 10 lg/mL), small interfering GAPDH 50-AAT CCC ATC ACC ATC TTC C-30 RNA (siRNA), or NAC (5 mM), RPE cells were tested for viability 50-TTG AGG CTG TTG TCA TAC TTC T-30 using LIVE/DEAD Fixable Near-IR Dead Cell Stain Kit (Invitro- gen, Carlsbad, CA, USA) and flow cytometry. The cell viability was above 90% and did not decrease following these treatments. local IL-17A may serve as a trigger for inflammasome signaling and IL-1b production in RPE cells, forming a positive feedback Immunofluorescence of RPE Cells loop. In the present study, we first aimed to examine the expression pattern of IL-17 receptor A (IL-17RA), IL-17RC, and The 50% confluent cultured RPE cells were fixed in PBS ACT1 in primary human RPE cells. We next characterized the containing 3.7% paraformaldehyde. They then were blocked putative role of IL-17A in the production of IL-1b and the with the appropriate normal serum in PBS for 1 hour at room activation of NLRP3 inflammasome machinery in RPE cells. temperature. They then were incubated at 48C overnight with anti-RPE65 (1:200; Abcam). After being washed in PBS, the sections were incubated further with FITC-conjugated anti- MATERIALS AND METHODS mouse IgG (eBioscience) for 45 minutes at 378C. Control sections were treated in the same way, but with no primary Reagents mouse anti-RPE65 monoclonal antibody.
Load more

Recommended publications
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • 1714 Gene Comprehensive Cancer Panel Enriched for Clinically Actionable Genes with Additional Biologically Relevant Genes 400-500X Average Coverage on Tumor
    xO GENE PANEL 1714 gene comprehensive cancer panel enriched for clinically actionable genes with additional biologically relevant genes 400-500x average coverage on tumor Genes A-C Genes D-F Genes G-I Genes J-L AATK ATAD2B BTG1 CDH7 CREM DACH1 EPHA1 FES G6PC3 HGF IL18RAP JADE1 LMO1 ABCA1 ATF1 BTG2 CDK1 CRHR1 DACH2 EPHA2 FEV G6PD HIF1A IL1R1 JAK1 LMO2 ABCB1 ATM BTG3 CDK10 CRK DAXX EPHA3 FGF1 GAB1 HIF1AN IL1R2 JAK2 LMO7 ABCB11 ATR BTK CDK11A CRKL DBH EPHA4 FGF10 GAB2 HIST1H1E IL1RAP JAK3 LMTK2 ABCB4 ATRX BTRC CDK11B CRLF2 DCC EPHA5 FGF11 GABPA HIST1H3B IL20RA JARID2 LMTK3 ABCC1 AURKA BUB1 CDK12 CRTC1 DCUN1D1 EPHA6 FGF12 GALNT12 HIST1H4E IL20RB JAZF1 LPHN2 ABCC2 AURKB BUB1B CDK13 CRTC2 DCUN1D2 EPHA7 FGF13 GATA1 HLA-A IL21R JMJD1C LPHN3 ABCG1 AURKC BUB3 CDK14 CRTC3 DDB2 EPHA8 FGF14 GATA2 HLA-B IL22RA1 JMJD4 LPP ABCG2 AXIN1 C11orf30 CDK15 CSF1 DDIT3 EPHB1 FGF16 GATA3 HLF IL22RA2 JMJD6 LRP1B ABI1 AXIN2 CACNA1C CDK16 CSF1R DDR1 EPHB2 FGF17 GATA5 HLTF IL23R JMJD7 LRP5 ABL1 AXL CACNA1S CDK17 CSF2RA DDR2 EPHB3 FGF18 GATA6 HMGA1 IL2RA JMJD8 LRP6 ABL2 B2M CACNB2 CDK18 CSF2RB DDX3X EPHB4 FGF19 GDNF HMGA2 IL2RB JUN LRRK2 ACE BABAM1 CADM2 CDK19 CSF3R DDX5 EPHB6 FGF2 GFI1 HMGCR IL2RG JUNB LSM1 ACSL6 BACH1 CALR CDK2 CSK DDX6 EPOR FGF20 GFI1B HNF1A IL3 JUND LTK ACTA2 BACH2 CAMTA1 CDK20 CSNK1D DEK ERBB2 FGF21 GFRA4 HNF1B IL3RA JUP LYL1 ACTC1 BAG4 CAPRIN2 CDK3 CSNK1E DHFR ERBB3 FGF22 GGCX HNRNPA3 IL4R KAT2A LYN ACVR1 BAI3 CARD10 CDK4 CTCF DHH ERBB4 FGF23 GHR HOXA10 IL5RA KAT2B LZTR1 ACVR1B BAP1 CARD11 CDK5 CTCFL DIAPH1 ERCC1 FGF3 GID4 HOXA11 IL6R KAT5 ACVR2A
    [Show full text]
  • IL-1Β Induces the Rapid Secretion of the Antimicrobial Protein IL-26 From
    Published June 24, 2019, doi:10.4049/jimmunol.1900318 The Journal of Immunology IL-1b Induces the Rapid Secretion of the Antimicrobial Protein IL-26 from Th17 Cells David I. Weiss,*,† Feiyang Ma,†,‡ Alexander A. Merleev,x Emanual Maverakis,x Michel Gilliet,{ Samuel J. Balin,* Bryan D. Bryson,‖ Maria Teresa Ochoa,# Matteo Pellegrini,*,‡ Barry R. Bloom,** and Robert L. Modlin*,†† Th17 cells play a critical role in the adaptive immune response against extracellular bacteria, and the possible mechanisms by which they can protect against infection are of particular interest. In this study, we describe, to our knowledge, a novel IL-1b dependent pathway for secretion of the antimicrobial peptide IL-26 from human Th17 cells that is independent of and more rapid than classical TCR activation. We find that IL-26 is secreted 3 hours after treating PBMCs with Mycobacterium leprae as compared with 48 hours for IFN-g and IL-17A. IL-1b was required for microbial ligand induction of IL-26 and was sufficient to stimulate IL-26 release from Th17 cells. Only IL-1RI+ Th17 cells responded to IL-1b, inducing an NF-kB–regulated transcriptome. Finally, supernatants from IL-1b–treated memory T cells killed Escherichia coli in an IL-26–dependent manner. These results identify a mechanism by which human IL-1RI+ “antimicrobial Th17 cells” can be rapidly activated by IL-1b as part of the innate immune response to produce IL-26 to kill extracellular bacteria. The Journal of Immunology, 2019, 203: 000–000. cells are crucial for effective host defense against a wide and neutrophils.
    [Show full text]
  • Proinflammatory T Helper Type 17 Cells Are Effective B-Cell Helpers
    Proinflammatory T helper type 17 cells are effective B-cell helpers Meike Mitsdoerffera, Youjin Leea, Anneli Jägera, Hye-Jung Kimb, Thomas Kornc, Jay K. Kollsd, Harvey Cantorb,1, Estelle Bettellie,1, and Vijay K. Kuchrooa,1 aCenter for Neurologic Diseases, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115; bDana-Farber Cancer Institute, Boston, MA 02115; cDepartment of Neurology, Technical University Munich, 81675 Munich, Germany; dLouisiana State University Health Sciences Center, New Orleans, LA 70112; and eBenaroya Research Institute, Seattle, WA 98101 Contributed by Harvey Cantor, July 1, 2010 (sent for review May 11, 2010) T helper type 17 (TH17) cells are highly proinflammatory effector collaboration. TH1 and TH2 cells are known to provide B-cell T cells that are characterized by the production of high amounts of help, and their signature cytokines IFN-γ and IL-4 induce class IL-17A, IL-17F, IL-21, and IL-22. Furthermore, TH17 cells have been switch recombination to IgG2a or IgG1 and IgE, respectively (9, associated with a number of autoimmune diseases. However, it is 10). Using IFN-γ and IL-4 reporter mice, Reinhardt et al. (11) not clear whether TH17 cells can also serve as effective helper cells. demonstrated that B cells form conjugates with antigen-activated, Here we show that TH17 cells can function as B-cell helpers in cytokine-producing effector T cells and that the profile of cyto- that they not only induce a strong proliferative response of B cells kine production defines isotype class switching in conjugated B in vitro but also trigger antibody production with class switch cells.
    [Show full text]
  • IL-17-Producing Cells in Tumor Immunity: Friends Or Foes?
    Immune Netw. 2020 Feb;20(1):e6 https://doi.org/10.4110/in.2020.20.e6 pISSN 1598-2629·eISSN 2092-6685 Review Article IL-17-Producing Cells in Tumor Immunity: Friends or Foes? Da-Sol Kuen 1,2, Byung-Seok Kim 1, Yeonseok Chung 1,2,* 1Laboratory of Immune Regulation, Institute of Pharmaceutical Sciences, Seoul National University, Seoul 08826, Korea 2BK21 Plus Program, Institute of Pharmaceutical Sciences, Seoul National University, Seoul 08826, Korea Received: Dec 29, 2019 Revised: Jan 25, 2020 ABSTRACT Accepted: Jan 26, 2020 IL-17 is produced by RAR-related orphan receptor gamma t (RORγt)-expressing cells *Correspondence to including Th17 cells, subsets of γδT cells and innate lymphoid cells (ILCs). The biological Yeonseok Chung significance of IL-17-producing cells is well-studied in contexts of inflammation, Laboratory of Immune Regulation and BK21 Plus Program, Institute of Pharmaceutical autoimmunity and host defense against infection. While most of available studies in tumor + Sciences, Seoul National University, 1 Gwanak- immunity mainly focused on the role of T-bet-expressing cells, including cytotoxic CD8 ro, Gwanak-gu, Seoul 08826, Korea. T cells and NK cells, and their exhaustion status, the role of IL-17-producing cells remains E-mail: [email protected] poorly understood. While IL-17-producing T-cells were shown to be anti-tumorigenic in Copyright © 2020. The Korean Association of adoptive T-cell therapy settings, mice deficient in type 17 genes suggest a protumorigenic Immunologists potential of IL-17-producing cells. This review discusses the features of IL-17-producing This is an Open Access article distributed cells, of both lymphocytic and myeloid origins, as well as their suggested pro- and/or anti- under the terms of the Creative Commons tumorigenic functions in an organ-dependent context.
    [Show full text]
  • Single-Cell Analysis of Crohn's Disease Lesions Identifies
    bioRxiv preprint doi: https://doi.org/10.1101/503102; this version posted December 20, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Single-cell analysis of Crohn’s disease lesions identifies a pathogenic cellular module associated with resistance to anti-TNF therapy JC Martin1,2,3, G Boschetti1,2,3, C Chang1,2,3, R Ungaro4, M Giri5, LS Chuang5, S Nayar5, A Greenstein6, M. Dubinsky7, L Walker1,2,5,8, A Leader1,2,3, JS Fine9, CE Whitehurst9, L Mbow9, S Kugathasan10, L.A. Denson11, J.Hyams12, JR Friedman13, P Desai13, HM Ko14, I Laface1,2,8, Guray Akturk1,2,8, EE Schadt15,16, S Gnjatic1,2,8, A Rahman1,2,5,8, , M Merad1,2,3,8,17,18*, JH Cho5,17,*, E Kenigsberg1,15,16,17* 1 Precision Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. 2 Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. 3 Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. 4 The Dr. Henry D. Janowitz Division of Gastroenterology, Icahn School of Medicine at Mount Sinai, New York City, NY 10029, USA. 5 Charles Bronfman Institute for Personalized Medicine, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. 6 Department of Colorectal Surgery, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA 7 Department of Pediatrics, Susan and Leonard Feinstein IBD Clinical Center, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
    [Show full text]
  • Evolutionary Divergence and Functions of the Human Interleukin (IL) Gene Family Chad Brocker,1 David Thompson,2 Akiko Matsumoto,1 Daniel W
    UPDATE ON GENE COMPLETIONS AND ANNOTATIONS Evolutionary divergence and functions of the human interleukin (IL) gene family Chad Brocker,1 David Thompson,2 Akiko Matsumoto,1 Daniel W. Nebert3* and Vasilis Vasiliou1 1Molecular Toxicology and Environmental Health Sciences Program, Department of Pharmaceutical Sciences, University of Colorado Denver, Aurora, CO 80045, USA 2Department of Clinical Pharmacy, University of Colorado Denver, Aurora, CO 80045, USA 3Department of Environmental Health and Center for Environmental Genetics (CEG), University of Cincinnati Medical Center, Cincinnati, OH 45267–0056, USA *Correspondence to: Tel: þ1 513 821 4664; Fax: þ1 513 558 0925; E-mail: [email protected]; [email protected] Date received (in revised form): 22nd September 2010 Abstract Cytokines play a very important role in nearly all aspects of inflammation and immunity. The term ‘interleukin’ (IL) has been used to describe a group of cytokines with complex immunomodulatory functions — including cell proliferation, maturation, migration and adhesion. These cytokines also play an important role in immune cell differentiation and activation. Determining the exact function of a particular cytokine is complicated by the influence of the producing cell type, the responding cell type and the phase of the immune response. ILs can also have pro- and anti-inflammatory effects, further complicating their characterisation. These molecules are under constant pressure to evolve due to continual competition between the host’s immune system and infecting organisms; as such, ILs have undergone significant evolution. This has resulted in little amino acid conservation between orthologous proteins, which further complicates the gene family organisation. Within the literature there are a number of overlapping nomenclature and classification systems derived from biological function, receptor-binding properties and originating cell type.
    [Show full text]
  • Peripheral and Islet Interleukin-17 Pathway Activation
    ORIGINAL ARTICLE Peripheral and Islet Interleukin-17 Pathway Activation Characterizes Human Autoimmune Diabetes and Promotes Cytokine-Mediated b-Cell Death Sefina Arif,1 Fabrice Moore,2 Katherine Marks,1 Thomas Bouckenooghe,2 Colin M. Dayan,3 Raquel Planas,4 Marta Vives-Pi,4 Jake Powrie,5,6 Timothy Tree,1 Piero Marchetti,7 Guo Cai Huang,8 Esteban N. Gurzov,7 Ricardo Pujol-Borrell,5 Decio L. Eizirik,2 and Mark Peakman1,6 OBJECTIVE—CD4 T-cells secreting interleukin (IL)-17 are implicated in several human autoimmune diseases, but their role in type 1 diabetes has not been defined. To address the relevance n organ-specific autoimmune diseases such as type 1 of such cells, we examined IL-17 secretion in response to b-cell diabetes, key pathological checkpoints are priming autoantigens, IL-17A gene expression in islets, and the potential and differentiation of T-cells specificforb-cell auto- functional consequences of IL-17 release for b-cells. antigens, migration of these autoreactive cells to the I – RESEARCH DESIGN AND METHODS—Peripheral blood CD4 islets of Langerhans, and b-cell selective death (1). The T-cell responses to b-cell autoantigens (proinsulin, insulinoma- recent identification of the importance of CD4 T-cells that associated protein, and GAD65 peptides) were measured by IL-17 secrete interleukin (IL)-17 in various rodent disease mod- enzyme-linked immunospot assay in patients with new-onset els (2,3) has focused attention on how T helper 17 (TH17) type 1 diabetes (n = 50). mRNA expression of IL-17A and IFNG effector cells might participate in critical disease pathways pathway genes was studied by qRT-PCR using islets obtained in humans.
    [Show full text]
  • Human IL17RC Protein (Fc Tag)
    Human IL17RC Protein (Fc Tag) Catalog Number: 11747-H02H General Information SDS-PAGE: Gene Name Synonym: IL17-RL; IL17RL; UNQ6118/PRO20040/PRO38901 Protein Construction: A DNA sequence encoding the extracellular domain of human IL17RC isoform 3 (NP_116121.2) (Met 1-Ala 454) was fused with the Fc region of human IgG1 at the C-terminus. Source: Human Expression Host: HEK293 Cells QC Testing Purity: > 95 % as determined by SDS-PAGE Bio Activity： Protein Description Measured by its binding ability in a functional ELISA. Immobilized IL17RC (Interleukin 17 Receptor C) is a Protein Coding gene. This gene Human IL17 (Cat:12047-HNAE) at 2 μg/ml (100 μl/well) can bind Human encodes a single-pass type I membrane protein that shares similarity with IL17RC hFc (Cat:11747-H02H), the EC50 of Human IL17RC hFc is 200-800 the interleukin-17 receptor (IL-17RA). IL17RC is widely expressed in the ng/mL. prostate, skin, and other tissues. The hypomethylation within the IL17RC gene promoter in peripheral blood is not suitable for use as a clinical Endotoxin: biomarker of AMD. This study highlights the need for considerable replication of epigenetic association studies before clinical application. < 1.0 EU per μg of the protein as determined by the LAL method methylation of IL17RC could play as a marker in choroi dal neovascularization (CNV) and degeneration of retinal pigment epithelium Leu 21 Predicted N terminal: (RPE) cells in vitro. Diseases associated with IL17RC include Candidiasis, Molecular Mass: Familial, 9, and Chronic Mucocutaneous Candidiasis. The secreted recombinant human IL17RC/Fc is a disulfide-linked homodimeric References protein.
    [Show full text]
  • Active Interleukin 17 Receptor C (IL17RC)
    APD350Hu01 100µg Active Interleukin 17 Receptor C (IL17RC) Organism Species: Homo sapiens (Human) Instruction manual FOR IN VITRO USE AND RESEARCH USE ONLY NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES 1st Edition (Apr, 2016) [ PROPERTIES ] Source: Prokaryotic expression. Host: E. coli Residues: Asn257~Asn477 Tags: N-terminal His-tag Purity: >98% Buffer Formulation: 20mM Tris, 150mM NaCl, pH8.0, containing 0.05% sarcosyl and 5% trehalose. Applications: Cell culture; Activity Assays; In vivo assays. (May be suitable for use in other assays to be determined by the end user.) Predicted isoelectric point: 5.8 Predicted Molecular Mass: 28.6kDa Accurate Molecular Mass: 26kDa as determined by SDS-PAGE reducing conditions. [ USAGE ] Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex. [ STORAGE AND STABILITY ] Storage: Avoid repeated freeze/thaw cycles. Store at 2-8oC for one month. Aliquot and store at -80oC for 12 months. Stability Test: The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37oC for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition. [ SEQUENCE ] [ ACTIVITY ] IL17RC (Interleukin-17 receptor C) is a receptor for IL17A and IL17F homodimers as part of a heterodimeric complex with IL17RA. This protein is expressed in nonhemopoietic tissues, and binds both IL-17A and IL-17F with similar affinities. A binding ELISA assay was conducted to detect the association of IL17RC with IL17RA.
    [Show full text]
  • IL17 (IL17A) Mouse Monoclonal Antibody [Clone ID: 4H1524.1] Product Data
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for TA319543 IL17 (IL17A) Mouse Monoclonal Antibody [Clone ID: 4H1524.1] Product data: Product Type: Primary Antibodies Clone Name: 4H1524.1 Applications: WB Recommend Dilution: WB: 1ug/mL, FC: User Optimized Reactivity: Human Host: Mouse Clonality: Monoclonal Immunogen: Anti-IL-17A (MOUSE) Monoclonal Antibody was produced in mouse by repeated immunizations with mature full length recombinant human IL-17A produced in E.coli followed by hybridoma development. Formulation: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 Concentration: 1 mg/ml Gene Name: interleukin 17A Database Link: NP_002181 Entrez Gene 3605 Human Synonyms: CTLA8; IL-17; IL-17A; IL17 Note: IL17-A (also known as Interleukin-17) is a proinflammatory cytokine produced by activated T cells. This cytokine regulates the activities of NF-kappaB and mitogen-activated protein kinases. This cytokine can stimulate the expression of IL6 and cyclooxygenase-2 (PTGS2/COX- 2), as well as enhance the production of nitric oxide (NO). High levels of this cytokine are associated with several chronic inflammatory diseases including rheumatoid arthritis, psoriasis and multiple sclerosis. IL17-A is the founding member of a group of cytokines called the IL17 family. IL17-A was originally identified as a transcript from a rodent T-cell hybridoma. To elicit its functions, IL17 binds to a type I cell surface receptor called IL17R of which there are at least three variants IL17RA, IL17RB, and IL17RC.
    [Show full text]
  • IL-17RA-Signaling Modulates CD8+ T Cell Survival and Exhaustion During 2 Trypanosoma Cruzi Infection
    bioRxiv preprint doi: https://doi.org/10.1101/314336; this version posted May 11, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-ND 4.0 International license. 1 IL-17RA-signaling modulates CD8+ T cell survival and exhaustion during 2 Trypanosoma cruzi infection 3 Jimena Tosello Boari1,2, Cintia L. Araujo Furlan1,2, Facundo Fiocca Vernengo1,2, Constanza 4 Rodriguez1,2, María C. Ramello1,2, María C. Amezcua Vesely1,2, Melisa Gorosito Serrán1,2, 5 Nicolás G. Nuñez3,4, Wilfrid Richer3,4, Eliane Piaggio3,4, Carolina L. Montes1,2, Adriana 6 Gruppi1,2, Eva V. Acosta Rodríguez1,2*. 7 1 Departamento de Bioquímica Clínica. Facultad de Ciencias Químicas, Universidad 8 Nacional de Córdoba, Córdoba, X5000HUA. Argentina. 9 2 Centro de Investigaciones en Bioquímica Clínica e Inmunología. CONICET. Córdoba, 10 X5000HUA. Argentina. 11 3 SiRIC TransImm «Translational Immunotherapy Team», Translational Research 12 Department, Research Center, PSL Research University, INSERM U932, Institut Curie, 13 Paris, 75005. France. 14 4 Centre d’Investigation Clinique Biothérapie CICBT 1428, Institut Curie, Paris, 75005. 15 France. 16 Running title: IL-17RA signaling regulates CD8+ T cell responses to T. cruzi 17 *Corresponding autor: 18 Eva V. Acosta Rodríguez 19 [email protected] 20 1 bioRxiv preprint doi: https://doi.org/10.1101/314336; this version posted May 11, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.
    [Show full text]