Glucocorticoids Suppress CCR9-Mediated Chemotaxis, Calcium Flux, and Adhesion to Madcam-1 in Human T Cells

Total Page:16

File Type:pdf, Size:1020Kb

Glucocorticoids Suppress CCR9-Mediated Chemotaxis, Calcium Flux, and Adhesion to Madcam-1 in Human T Cells Glucocorticoids Suppress CCR9-Mediated Chemotaxis, Calcium Flux, and Adhesion to MAdCAM-1 in Human T Cells This information is current as Emily Wendt, Gemma E. White, Helen Ferry, Michael of September 25, 2021. Huhn, David R. Greaves and Satish Keshav J Immunol published online 25 March 2016 http://www.jimmunol.org/content/early/2016/03/24/jimmun ol.1500619 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2016/03/24/jimmunol.150061 Material 9.DCSupplemental http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 25, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2016 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published March 25, 2016, doi:10.4049/jimmunol.1500619 The Journal of Immunology Glucocorticoids Suppress CCR9-Mediated Chemotaxis, Calcium Flux, and Adhesion to MAdCAM-1 in Human T Cells Emily Wendt,* Gemma E. White,† Helen Ferry,‡ Michael Huhn,* David R. Greaves,† and Satish Keshav* CCR9 expressed on T lymphocytes mediates migration to the small intestine in response to a gradient of CCL25. CCL25-stimulated activation of a4b7 integrin promotes cell adherence to mucosal addressin cell adhesion molecule-1 (MAdCAM-1) expressed by vascular endothelial cells of the intestine, further mediating gut-specific homing. Inflammatory bowel disease is a chronic inflam- matory condition that primarily affects the gastrointestinal tract and is characterized by leukocyte infiltration. Glucocorticoids (GCs) are widely used to treat inflammatory bowel disease but their effect on intestinal leukocyte homing is not well understood. Downloaded from We investigated the effect of GCs on the gut-specific chemokine receptor pair, CCR9 and CCL25. Using human peripheral blood- derived T lymphocytes enriched for CCR9 by cell sorting or culturing with all-trans retinoic acid, we measured chemotaxis, intracellular calcium flux, and a4b7-mediated cell adhesion to plate-bound MAdCAM-1. Dexamethasone (DEX), a specific GC receptor agonist, significantly reduced CCR9-mediated chemotaxis and adhesion to MAdCAM-1 without affecting CCR9 surface expression. In contrast, in the same cells, DEX increased CXCR4 surface expression and CXCL12-mediated signaling and downstream functions. The effects of DEX on human primary T cells were reversed by the GC receptor antagonist mifepristone. http://www.jimmunol.org/ These results demonstrate that GCs suppress CCR9-mediated chemotaxis, intracellular calcium flux, and a4b7-mediated cell adhesion in vitro, and these effects could contribute to the efficacy of GCs in treating intestinal inflammation in vivo. The Journal of Immunology, 2016, 196: 000–000. he chemokine CCL25 is constitutively expressed in the peripheral blood, the frequency of circulating CCR9+ T cells is small intestine and interacts with a single signaling re- elevated during small intestinal Crohn’s disease, suggesting a role T ceptor, CCR9 (1, 2). In humans, CCR9 is expressed on of CCR9+ T cells in small intestinal inflammation (11). CCR9, ∼ most (58–97%) T cells in the small intestine, 25% in the colon, CCL25, a4b7, and MAdCAM-1 are attractive therapeutic targets and a small minority (3–5%) in peripheral blood (2, 3). CCR9+ in IBD because of their tissue and cell specificity, and clinical by guest on September 25, 2021 T cells coexpress integrin a4b7, which binds to mucosal addressin studies show positive effects on blocking these proteins in subsets cell adhesion molecule-1 (MAdCAM-1) (4). MAdCAM-1 is consti- of IBD patients (12–15). tutively expressed on vascular endothelial cells in the small and Glucocorticoids (GCs) have broad-ranging anti-inflammatory large intestine, and together CCR9 and a4b7 preferentially me- actions, are highly effective in many chronic inflammatory dis- diate the adherence and migration of effector T cells into the orders, and have been the mainstay of IBD treatment since the intestine (5, 6). 1950s (16, 17). The anti-inflammatory effects of GCs are mainly Inflammatory bowel disease (IBD) is characterized by leukocyte mediated through interaction with cytosolic GC receptors (GCRs) infiltration, increased levels of proinflammatory cytokines in the to modify the transcription of proinflammatory genes (18, 19). intestine, and increased expression of MAdCAM-1 (7-10). In the Previously characterized effects of GCs on T cells include inhibiting TCR signaling, suppressing cell proliferation, inducing *Translational Gastroenterology Unit, Nuffield Department of Medicine, John Radcliffe IkB synthesis, and subsequently reducing NF-kB–mediated tran- † Hospital, Oxford OX3 9DU, United Kingdom; Sir William Dunn School of Pathology, scription of proinflammatory cytokines (19–21). However, little is University of Oxford, Oxford OX1 3RE, United Kingdom; and ‡Experimental Medicine Division, Nuffield Department of Medicine, University of Oxford, Oxford OX3 9DU, known about the effects of GCs on intestine-specific cell recruit- United Kingdom ment. We investigated the effect of GCs on CCR9 expression and Received for publication April 1, 2015. Accepted for publication February 22, 2016. function, as well as downstream a4b7 activation in primary human This work was supported by the Oxford Biomedical Research Centre, funded by the T cells, in vitro. United Kingdom National Institutes for Health Research Grant A93081 (to S.K.), as In this study we demonstrate that CCR9-mediated chemotaxis, well as by an unrestricted grant from ChemoCentryx, Inc. (Mountain View, CA). The funders had no role in the study design, data collection and analysis, decision to intracellular calcium flux, and adhesion to MAdCAM-1 are sig- publish, or preparation of the manuscript. nificantly suppressed following corticosteroid treatment, and this is Address correspondence and reprint requests to Dr. Satish Keshav, Gastroenterology mediated through activation of GCRs. As a control, we compared Unit, Nuffield Department of Medicine, Level 5, John Radcliffe Hospital, Headley + Way, Oxford OX3 9DU, U.K. E-mail address: [email protected] the effects of GCs on CXCR4, which is coexpressed on CCR9 The online version of this article contains supplemental material. T cells. CXCR4 is stimulated by a single ligand, CXCL12, and consistent with previous reports, GCs increase CXCR4 expression Abbreviations used in this article: ATRA, all-trans retinoic acid; DEX, dexamethasone; DOC, deoxycorticosterone acetate; GC, glucocorticoid; GCR, glucocorticoid receptor; and enhance CXCL12-mediated functions (22, 23). These results IBD, inflammatory bowel disease; MAdCAM-1, mucosal addressin cell adhesion mole- identify novel effects of GCs on the function of primary human cule-1; MFI, mean fluorescence intensity; MIF, mifepristone; PRED, prednisolone. T cells that are likely to be physiologically and clinically relevant Copyright Ó 2016 by The American Association of Immunologists, Inc. 0022-1767/16/$30.00 in intestinal inflammation. www.jimmunol.org/cgi/doi/10.4049/jimmunol.1500619 2 GLUCOCORTICOIDS SUPPRESS CCR9 FUNCTION Materials and Methods Calcium mobilization assay All-trans retinoic acid–cultured cells Vehicle (H2O) and DEX-treated cells were separately labeled with anti- PBMCs were depleted of monocytes by CD14+ bead selection (Miltenyi CCR9 (R&D Systems, clone 248621) followed by anti-mouse IgG con- Biotec, Surrey, U.K.). Remaining leukocytes were activated by culturing on jugated to allophycocyanin in PBS with 0.1% BSA. Vehicle-treated cells plate-bound anti-CD3 (3 mg/ml) (BioLegend, Cambridge, U.K., clone OKT3) were labeled with anti-CD3 Alexa Fluor 700 and DEX-treated cells with and anti-CD28 (3 mg/ml) (BioLegend, clone CD28.2) in complete culture anti-CD3 allophycocyanin–eFluor 780 (eBioscience, clone UCHT1). An medium (RPMI 1640 supplemented with 10% FCS, penicillin-streptomycin) equivalent number of vehicle- and DEX-treated cells were combined in with 100 U IL-2 (PeproTech, Rocky Hill, NJ) and 100 nM all-trans retinoic HBSS with 0.02% Pluronic F-127, 1 mg/ml Fura Red AM (Life Tech- acid (ATRA). After 72 h, cells were removed from CD3/CD28 stimulation and nologies), and incubated for 30 min at 37˚C. Cells were resuspended in resuspended in complete culture medium with IL-2 and ATRA. Cells were HBSS with 0.1% BSA, 1 mM CaCl2, 0.5 mM MgCl2, and 10 mM HEPES used for assays between days 6 and 9 from initial isolation. with Live/Dead cell marker, SYTOX Green (Life Technologies), and in- cubated at 37˚C until flow cytometry was performed. Data were acquired on an LSR II SORP and analysis was performed in FlowJo. The gating Steroid and steroid receptor antagonist treatment 2 strategy was as follows: live cells (SYTOX Green ), excluding cell dou- Cells were resuspended in RPMI 1640 supplemented with 10% FCS, blets (determined by forward light scatter area versus forward light scatter 50 U/ml penicillin, and 50 mg/ml streptomycin. Cells were
Recommended publications
  • Disease Lymphocytes in Small Intestinal Crohn's Chemokine
    Phenotype and Effector Function of CC Chemokine Receptor 9-Expressing Lymphocytes in Small Intestinal Crohn's Disease This information is current as of September 29, 2021. Masayuki Saruta, Qi T. Yu, Armine Avanesyan, Phillip R. Fleshner, Stephan R. Targan and Konstantinos A. Papadakis J Immunol 2007; 178:3293-3300; ; doi: 10.4049/jimmunol.178.5.3293 http://www.jimmunol.org/content/178/5/3293 Downloaded from References This article cites 26 articles, 12 of which you can access for free at: http://www.jimmunol.org/content/178/5/3293.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 29, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2007 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Phenotype and Effector Function of CC Chemokine Receptor 9-Expressing Lymphocytes in Small Intestinal Crohn’s Disease1 Masayuki Saruta,2*QiT.Yu,2* Armine Avanesyan,* Phillip R. Fleshner,† Stephan R.
    [Show full text]
  • Atypical Chemokine Receptor 4 Shapes Activated B Cell Fate
    Brief Definitive Report Atypical chemokine receptor 4 shapes activated B cell fate Ervin E. Kara,1 Cameron R. Bastow,1 Duncan R. McKenzie,1 Carly E. Gregor,1 Kevin A. Fenix,1 Rachelle Babb,1 Todd S. Norton,1 Dimitra Zotos,3 Lauren B. Rodda,4 Jana R. Hermes,6 Katherine Bourne,6 Derek S. Gilchrist,7 Robert J. Nibbs,7 Mohammed Alsharifi,1 Carola G. Vinuesa,8 David M. Tarlinton,3,9 Robert Brink,6,10 Geoffrey R. Hill,11 Jason G. Cyster,4,5 Iain Comerford,1 and Shaun R. McColl1,2 1Department of Molecular and Cellular Biology, School of Biological Sciences and 2Centre for Molecular Pathology, School of Biological Sciences, University of Adelaide, Adelaide, South Australia, Australia 3Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia 4Department of Microbiology and Immunology and 5Howard Hughes Medical Institute, Department of Microbiology and Immunology, University of California, Downloaded from http://rupress.org/jem/article-pdf/215/3/801/1168927/jem_20171067.pdf by guest on 28 September 2021 San Francisco, San Francisco, CA 6Immunology Division, Garvan Institute of Medical Research, Darlinghurst, New South Wales, Australia 7Institute of Infection, Immunity and Inflammation, College of Medicine, Veterinary and Life Sciences, University of Glasgow, Glasgow, Scotland, UK 8Department of Immunology and Infectious Disease, John Curtin School of Medical Research, Australian National University, Canberra, Australian Capital Territory, Australia 9Department of Immunology and Pathology, Monash University, Melbourne, Victoria, Australia 10St Vincent’s Clinical School, University of New South Wales, Darlinghurst, New South Wales, Australia 11Immunology Department, QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia Activated B cells can initially differentiate into three functionally distinct fates—early plasmablasts (PBs), germinal center (GC) B cells, or early memory B cells—by mechanisms that remain poorly understood.
    [Show full text]
  • Chemokine Receptor CXCR3 Promotes Colon Cancer Metastasis to Lymph Nodes
    Oncogene (2007) 26, 4679–4688 & 2007 Nature Publishing Group All rights reserved 0950-9232/07 $30.00 www.nature.com/onc ORIGINAL ARTICLE Chemokine receptor CXCR3 promotes colon cancer metastasis to lymph nodes K Kawada1,2,5, H Hosogi1,2,5, M Sonoshita1, H Sakashita3, T Manabe3, Y Shimahara2, Y Sakai2, A Takabayashi4, M Oshima1 and MM Taketo1 1Department of Pharmacology, Graduate School of Medicine, Kyoto University, Kyoto, Japan; 2Department of Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan; 3Department of Clinical Pathology, Graduate School of Medicine, Kyoto University, Kyoto, Japan and 4Kitano Hospital Medical Institute, Osaka, Japan Chemokines and their receptors are essential for leuko- inflammatory cytokines, growth factors and/or patho- cyte trafficking, and also implicated in cancer metastasis genic stimuli. Important roles of chemokines and their to specific organs. We have recently demonstrated that receptors have been demonstrated in inflammation, CXCR3 plays a critical role in metastasis of mouse infection, tissue injury, allergy and cardiovascular melanoma cells to lymph nodes. Here, we show that some diseases as well as in malignant tumors. Chemokine human colon cancer cell lines express CXCR3 constitu- receptor CXCR3 is essential for the physiologic and tively. We constructed cells that expressed CXCR3 cDNA pathologic recruitment of plasmacytoid dendritic cell (‘DLD-1-CXCR3’), and compared with nonexpressing precursors, monocytes and natural killer cells to controls by rectal transplantation in nude mice. Although inflamed lymph nodes (LNs) (Cella et al., 1999; both cell lines disseminated to lymph nodes at similar Janatpour et al., 2001; Martin-Fontecha et al., 2004), frequencies at 2 weeks, DLD-1-CXCR3 expanded more and for retention of Th1 lymphocytes within LNs rapidly than the control in 4 weeks.
    [Show full text]
  • G Protein-Coupled Receptors As Therapeutic Targets for Multiple Sclerosis
    npg GPCRs as therapeutic targets for MS Cell Research (2012) 22:1108-1128. 1108 © 2012 IBCB, SIBS, CAS All rights reserved 1001-0602/12 $ 32.00 npg REVIEW www.nature.com/cr G protein-coupled receptors as therapeutic targets for multiple sclerosis Changsheng Du1, Xin Xie1, 2 1Laboratory of Receptor-Based BioMedicine, Shanghai Key Laboratory of Signaling and Disease Research, School of Life Sci- ences and Technology, Tongji University, Shanghai 200092, China; 2State Key Laboratory of Drug Research, the National Center for Drug Screening, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 189 Guo Shou Jing Road, Pudong New District, Shanghai 201203, China G protein-coupled receptors (GPCRs) mediate most of our physiological responses to hormones, neurotransmit- ters and environmental stimulants. They are considered as the most successful therapeutic targets for a broad spec- trum of diseases. Multiple sclerosis (MS) is an inflammatory disease that is characterized by immune-mediated de- myelination and degeneration of the central nervous system (CNS). It is the leading cause of non-traumatic disability in young adults. Great progress has been made over the past few decades in understanding the pathogenesis of MS. Numerous data from animal and clinical studies indicate that many GPCRs are critically involved in various aspects of MS pathogenesis, including antigen presentation, cytokine production, T-cell differentiation, T-cell proliferation, T-cell invasion, etc. In this review, we summarize the recent findings regarding the expression or functional changes of GPCRs in MS patients or animal models, and the influences of GPCRs on disease severity upon genetic or phar- macological manipulations.
    [Show full text]
  • A Subset of CCL25-Induced Gut-Homing T Cells Affects Intestinal Immunity to Infection and Cancer
    ORIGINAL RESEARCH published: 25 February 2019 doi: 10.3389/fimmu.2019.00271 A Subset of CCL25-Induced Gut-Homing T Cells Affects Intestinal Immunity to Infection and Cancer Hongmei Fu 1, Maryam Jangani 1, Aleesha Parmar 1, Guosu Wang 1, David Coe 1, Sarah Spear 2†, Inga Sandrock 3, Melania Capasso 2†, Mark Coles 4, Georgina Cornish 1, Helena Helmby 5 and Federica M. Marelli-Berg 1* 1 William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom, 2 Bart’s Cancer Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom, 3 Institute of Immunology, Hannover Medical School, Hannover, 4 5 Edited by: Germany, Kennedy Institute of Rheumatology, University of Oxford, Oxford, United Kingdom, Department for Immunology Mariagrazia Uguccioni, and Infection, London School of Hygiene and Tropical Medicine, London, United Kingdom Institute for Research in Biomedicine (IRB), Switzerland Protective immunity relies upon differentiation of T cells into the appropriate subtype Reviewed by: required to clear infections and efficient effector T cell localization to antigen-rich tissue. Maria Rescigno, Istituto Europeo di Oncologia s.r.l., Recent studies have highlighted the role played by subpopulations of tissue-resident Italy memory (TRM) T lymphocytes in the protection from invading pathogens. The intestinal Fabio Grassi, Institute for Research in Biomedicine mucosa and associated lymphoid tissue are densely populated by a variety of resident (IRB), Switzerland lymphocyte populations, including αβ and γδ CD8+ intraepithelial T lymphocytes (IELs) *Correspondence: and CD4+ T cells. While the development of intestinal γδ CD8+ IELs has been extensively Federica M.
    [Show full text]
  • CC Chemokine Ligand 25 Enhances Resistance to Apoptosis in CD4 T
    [CANCER RESEARCH 64, 7579–7587, October 15, 2004] CC Chemokine Ligand 25 Enhances Resistance to Apoptosis in CD4؉ T Cells from Patients with T-Cell Lineage Acute and Chronic Lymphocytic Leukemia by Means of Livin Activation Zhang Qiuping,1 Xiong Jei,1,2 Jin Youxin,2 Ju Wei,1 Liu Chun,1 Wang Jin,1 Wu Qun,1 Liu Yan,1 Hu Chunsong,3 Yang Mingzhen,4 Gao Qingping,5 Zhang Kejian,5 Sun Zhimin,6 Li Qun,3 Liu Junyan,1 and Tan Jinquan1,3 1Department of Immunology, and Laboratory of Allergy and Clinical Immunology, Institute of Allergy and Immune-related Diseases and Center for Medical Research, Wuhan University School of Medicine, Wuhan; 2The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Science, Shanghai; 3Department of Immunology, College of Basic Medical Sciences, Anhui Medical University, Hefei; 4Department of Hematology, The Affiliated University Hospital, Anhui Medical University, Hefei; 5Department of Hematology, The First and Second Affiliated University Hospital, Wuhan University, Wuhan; and 6Department of Hematology, The Provincial Hospital of Anhui, Hefei, Peoples Republic of China ABSTRACT intestine (8), providing the evidence for distinctive mechanisms of -؉ lymphocyte recruitment. The importance of CCL25/TECK is to li We investigated CD4 and CD8 double-positive thymocytes, CD4 T cense effector/memory cells to access anatomic sites (9, 10). Thus, cells from typical patients with T-cell lineage acute lymphocytic leukemia CCL25/TECK is important for the homing, development, and home- (T-ALL) and T cell lineage chronic lymphocytic leukemia (T-CLL), and MOLT4 T cells in terms of CC chemokine ligand 25 (CCL25) functions of ostasis of T cells, particularly, mucosal T cells.
    [Show full text]
  • Th2 Effector Lymphocytes Regulatory and + Cells Enriched for FOXP3
    CCR8 Expression Identifies CD4 Memory T Cells Enriched for FOXP3 + Regulatory and Th2 Effector Lymphocytes This information is current as Dulce Soler, Tobias R. Chapman, Louis R. Poisson, Lin of September 27, 2021. Wang, Javier Cote-Sierra, Mark Ryan, Alice McDonald, Sunita Badola, Eric Fedyk, Anthony J. Coyle, Martin R. Hodge and Roland Kolbeck J Immunol 2006; 177:6940-6951; ; doi: 10.4049/jimmunol.177.10.6940 Downloaded from http://www.jimmunol.org/content/177/10/6940 References This article cites 68 articles, 27 of which you can access for free at: http://www.jimmunol.org/content/177/10/6940.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 27, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2006 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology CCR8 Expression Identifies CD4 Memory T Cells Enriched for FOXP3؉ Regulatory and Th2 Effector Lymphocytes Dulce Soler,1 Tobias R.
    [Show full text]
  • Mouse CCL25/TECK Antibody
    Mouse CCL25/TECK Antibody Monoclonal Rat IgG2A Clone # 89827 Catalog Number: MAB4811 DESCRIPTION Species Reactivity Mouse Specificity Detects mouse CCL25/TECK in ELISAs and Western blots. In Western blots, no cross­reactivity with recombinant human CCL1, 2, 3, 4, 5, 7, 8, 11, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, recombinant mouse CCL1, 2, 3, 4, 6, 7, 9, 11, 12, 19, 20, 21, 22, 24, and recombinant rat CCL20 is observed. Source Monoclonal Rat IgG2A Clone # 89827 Purification Protein A or G purified from hybridoma culture supernatant Immunogen E. coli­derived recombinant mouse CCL25/TECK Gln24­Asn144 Accession # O35903.1 Endotoxin Level <0.10 EU per 1 μg of the antibody by the LAL method. Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details. *Small pack size (­SP) is supplied either lyophilized or as a 0.2 μm filtered solution in PBS. APPLICATIONS Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website. Recommended Sample Concentration Western Blot 1 µg/mL Recombinant Mouse CCL25/TECK (Catalog # 481­TK) Immunohistochemistry 8­25 µg/mL Perfusion fixed frozen sections of mouse intestine and perfusion fixed frozen sections of rat intestine Mouse CCL25/TECK Sandwich Immunoassay Reagent ELISA Capture 2­8 µg/mL Mouse CCL25/TECK Antibody (Catalog # MAB4811) ELISA Detection 0.1­0.4 µg/mL Mouse CCL25/TECK Biotinylated Antibody (Catalog # BAF481) Standard Recombinant Mouse CCL25/TECK (Catalog # 481­TK) PREPARATION AND STORAGE Reconstitution Reconstitute at 0.5 mg/mL in sterile PBS.
    [Show full text]
  • Expression of the Atypical Chemokine Receptor ACKR4 Identifies a Novel
    Expression of the Atypical Chemokine Receptor ACKR4 Identifies a Novel Population of Intestinal Submucosal Fibroblasts That Preferentially Expresses This information is current as Endothelial Cell Regulators of September 26, 2021. Carolyn A. Thomson, Serge A. van de Pavert, Michelle Stakenborg, Evelien Labeeuw, Gianluca Matteoli, Allan McI Mowat and Robert J. B. Nibbs J Immunol published online 14 May 2018 Downloaded from http://www.jimmunol.org/content/early/2018/05/11/jimmun ol.1700967 http://www.jimmunol.org/ Supplementary http://www.jimmunol.org/content/suppl/2018/05/11/jimmunol.170096 Material 7.DCSupplemental Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on September 26, 2021 • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2018 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published May 14, 2018, doi:10.4049/jimmunol.1700967 The Journal of Immunology Expression of the Atypical Chemokine Receptor ACKR4 Identifies a Novel Population of Intestinal Submucosal Fibroblasts That Preferentially Expresses Endothelial Cell Regulators Carolyn A. Thomson,*,1 Serge A.
    [Show full text]
  • The Chemokine System in Innate Immunity
    Downloaded from http://cshperspectives.cshlp.org/ on September 28, 2021 - Published by Cold Spring Harbor Laboratory Press The Chemokine System in Innate Immunity Caroline L. Sokol and Andrew D. Luster Center for Immunology & Inflammatory Diseases, Division of Rheumatology, Allergy and Immunology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02114 Correspondence: [email protected] Chemokines are chemotactic cytokines that control the migration and positioning of immune cells in tissues and are critical for the function of the innate immune system. Chemokines control the release of innate immune cells from the bone marrow during homeostasis as well as in response to infection and inflammation. Theyalso recruit innate immune effectors out of the circulation and into the tissue where, in collaboration with other chemoattractants, they guide these cells to the very sites of tissue injury. Chemokine function is also critical for the positioning of innate immune sentinels in peripheral tissue and then, following innate immune activation, guiding these activated cells to the draining lymph node to initiate and imprint an adaptive immune response. In this review, we will highlight recent advances in understanding how chemokine function regulates the movement and positioning of innate immune cells at homeostasis and in response to acute inflammation, and then we will review how chemokine-mediated innate immune cell trafficking plays an essential role in linking the innate and adaptive immune responses. hemokines are chemotactic cytokines that with emphasis placed on its role in the innate Ccontrol cell migration and cell positioning immune system. throughout development, homeostasis, and in- flammation. The immune system, which is de- pendent on the coordinated migration of cells, CHEMOKINES AND CHEMOKINE RECEPTORS is particularly dependent on chemokines for its function.
    [Show full text]
  • Characterization of Mononuclear Phagocytic Cells in Medaka Fish
    Characterization of mononuclear phagocytic cells in medaka fish transgenic for a cxcr3a:gfp reporter Narges Aghaallaeia,1, Baubak Bajoghlia,1, Heinz Schwarzb, Michael Schorppa, and Thomas Boehma,2 aDepartment of Developmental Immunology, Max-Planck Institute of Immunobiology, D-79108 Freiburg, Germany; and bMax-Planck Institute for Developmental Biology, D-72076 Tübingen, Germany Edited* by Laurie H. Glimcher, Harvard University, Boston, MA, and approved September 10, 2010 (received for review January 13, 2010) Chemokines and chemokine receptors are key evolutionary innova- vertebrates possess many cell types that are characteristic of the tions of vertebrates. They are involved in morphogenetic processes adaptive and innate immune systems of mammals. The sites of or- and play an important role in the immune system. Based on an igin and developmental pathways of lymphocytes (1, 16, 17), as well analysis of the chemokine receptor gene family in teleost genomes, as those of the myeloid lineages (macrophages, neutrophils) (18– and the expression patterns of chemokine receptor genes during 22), have been well characterized. This was achieved by use of cell embryogenesis and the wounding response in young larvae of Ory- type-specific antibodies recognizing characteristic and evolution- zias latipes,weidentified the chemokine receptor cxcr3a as a marker arily conserved cell surface molecules or, for some species—most of innate immune cells. Cells expressing cxcr3a were characterized notably zebrafish—by use of transgenic fish in which the expression in fish transgenic for a cxcr3a:gfp reporter. In embryos and larvae, of fluorescent reporter proteins is directed by the regulatory regions cxcr3a-expressing cells are motile in healthy and damaged tissues, of specific genes to allow the in vivo visualization and preparative and phagocytic; the majority of these cells has the morphology of recovery of individual cell types.
    [Show full text]
  • Polyclonal Anti-CCR1 Antibody
    FabGennix International, Inc. 9191 Kyser Way Bldg. 4 Suite 402 Frisco, TX 75033 Tel: (214)-387-8105, 1-800-786-1236 Fax: (214)-387-8105 Email: [email protected] Web: www.FabGennix.com Polyclonal Anti-CCR1 antibody Catalog Number: CCR1-112AP General Information Product CCR1 Antibody Affinity Purified Description Chemokine (C-C motif) receptor 1 Antibody Affinity Purified Accession # Uniprot: P32246 GenBank: AAH64991 Verified Applications ELISA, WB Species Cross Reactivity Human Host Rabbit Immunogen Synthetic peptide taken within amino acid region 1-50 on human CCR1 protein. Alternative Nomenclature C C chemokine receptor type 1 antibody, C C CKR 1 antibody, CCR1 antibody, CD191 antibody, CMKBR 1 antibody, CMKR1 antibody, HM145 antibody, LD78 receptor antibody, Macrophage inflammatory protein 1 alpha /Rantes receptor antibody, MIP-1alpha-R antibody, MIP1aR antibody, RANTES receptor antibody, SCYAR1 antibody Physical Properties Quantity 100 µg Volume 200 µl Form Affinity Purified Immunoglobulins Immunoglobulin & Concentration 0.65-0.75 mg/ml IgG in antibody stabilization buffer Storage Store at -20⁰C for long term storage. Recommended Dilutions DOT Blot 1:10,000 ELISA 1:10,000 Western Blot 1:500 Related Products Catalog # FITC-Conjugated CCR1.112-FITC Antigenic Blocking Peptide P-CCR1.112 Western Blot Positive Control PC-CCR1.112 Tel: (214)-387-8105, 1-800-786-1236 Fax: (214)-387-8105 Email: [email protected] Web: www.FabGennix.com Overview: Chemokine receptors represent a subfamily of ~20 GPCRs that were originally identified by their roles in immune cell trafficking. Macrophage inflammatory protein-1 alpha (MIP-1 alpha) and RANTES, members of the beta chemokine family of leukocyte chemo- attractants, bind to a common seven-transmembrane-domain human receptor.
    [Show full text]