DOCSLIB.ORG

UBD, a Downstream Element of FOXP3, Allows the Identification of LGALS3, a New Marker of Human Regulatory T Cells

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
UBD, a Downstream Element of FOXP3, Allows the Identification of LGALS3, a New Marker of Human Regulatory T Cells Laboratory Investigation (2006) 86, 724–737 & 2006 USCAP, Inc All rights reserved 0023-6837/06 $30.00 www.laboratoryinvestigation.org UBD, a downstream element of FOXP3, allows the identification of LGALS3, a new marker of human regulatory T cells Frank Ocklenburg1,*, Darius Moharregh-Khiabani1,*, Robert Geffers2, Viktoria Janke1, Susanne Pfoertner2, Henk Garritsen3, Lothar Groebe2, Juergen Klempnauer1, Kurt EJ Dittmar4, Siegfried Weiss5, Jan Buer2,6 and Michael Probst-Kepper1 1Junior Research Group for Xenotransplantation, Department of Visceral and Transplant Surgery, Hannover Medical School, Hannover, Germany; 2Mucosal Immunity Research Group, German Research Centre for Biotechnology (GBF), Braunschweig, Germany; 3Institute for Clinical Transfusion Medicine, Sta¨dtisches Klinikum Braunschweig gGmbH, Braunschweig, Germany; 4Department of Molecular Biotechnology, German Research Centre for Biotechnology (GBF), Braunschweig, Germany; 5Department fo Molecular Immunology, German Research Centre for Biotechnology (GBF), Braunschweig, Germany and 6Insitute of Medical Microbiology, Hannover Medical School, Hannover, Germany Here, we report the identification of the ubiquitin-like gene UBD as a downstream element of FOXP3 in human þ hi activated regulatory CD4 CD25 T cells (Treg). Retroviral transduction of UBD in human allo-reactive effector þ CD4 T helper (Th) cells upregulates CD25 and mediates downregulation of IL4 and IL5 expression similar to overexpression of FOXP3. Moreover, UBD impairs Th cell proliferation without upregulation of FOXP3 and impairs calcium mobilization. In the presence of ionomycin, overexpression of UBD in Th cells leads to the induction of IL1R2 that resemble FOXP3-transduced Th cells and naturally derived Treg cells. A comparison of the transcriptome of FOXP3- and UBD-transduced Th cells with Treg cells allowed the identification of the gene LGALS3. However, high levels of LGALS3 protein expression were observed only in human CD4 þ CD25hi derived Treg cells and FOXP3-transduced Th cells, whereas little was induced in UBD-transduced Th cells. Thus, UBD contributes to the anergic phenotype of human regulatory T cells and acts downstream in FOXP3 induced regulatory signaling pathways, including regulation of LGALS3 expression. High levels of LGALS3 expression represent a FOXP3-signature of human antigen-stimulated CD4 þ CD25hi derived regulatory T cells. Laboratory Investigation (2006) 86, 724–737. doi:10.1038/labinvest.3700432; published online 15 May 2006 Keywords: CTLA4; FOXP3; LGALS3; regulatory T cells; UBD; CD4 þ CD25hi þ þ Regulatory CD4 CD25 T(Treg) cells have emerged been illustrated in different human disease settings, as a unique population of suppressor T cells that are that is, altered numbers and impairment of suppres- important for the maintenance of peripheral toler- sor activity of Treg cells has been demonstrated in 1,2 ance. In humans, Treg cells are characterized by patients with Hodgkin’s lymphomas, autoimmune high levels of CD25 expression (CD25hi) and repre- polyglandular syndrome type II, multiple sclerosis, sent 1–3% of all CD4 þ T cells in peripheral blood.3 myasthenia gravis, diabetes, psoriasis, patients with Typically, such Treg cells are anergic upon stimula- graft-versus-host-disease, and infectious diseases tion via their T-cell receptor and exhibit suppressive such as malaria and HIV.4–12 2 activity towards effector T-cell function. The clin- Development and function of Treg cells critically þ hi ical relevance of CD4 CD25 Treg cells has recently depend on the forkhead transcription factor FOXP3 that specifies the regulatory T cell lineage.13 As a result, humans and mice deficient in FOXP3 lack Correspondence: Dr M Probst-Kepper, MD, Department of Visceral þ þ and Transplant Surgery, Hannover Medical School, Building K25, CD4 CD25 Treg cells and suffer from multiorgan 2nd Floor, Room 1140, Carl Neuberg Strasse 1, D-30625 Hannover, autoimmune disease.2 Germany. Treg cells normally develop in the thymus. How- E-mail: [email protected] ever, they also may arise in a FOXP3-dependent *These authors contributed equally to this work. þ À 14,15 Received 25 January 2006; revised 31 March 2006; accepted 10 manner from peripheral CD4 CD25 T cells. The April 2006; published online 15 May 2006 antigen-specificity of Treg cells is currently a matter UBD and LGALS3 represent FOXP3-dependent genes F Ocklenburg et al 725 of debate, although self-antigens most likely repre- b-galactoside binding protein LGALS3. High levels sent the dominating specificity.16–19 of LGALS3 represent a FOXP3-signature of human þ hi Mechanisms of immunosuppression by Treg cells antigen-stimulated CD4 CD25 derived Treg cells as comprise cell-contact dependent suppression, it is not upregulated in nonregulatory CD4 þ CD25À mainly observed in vitro, and cytokine-mediated derived effector Th cells despite the expression of suppression of effector T cells, mainly observed in CD25 and FOXP3. vivo.20 Cytokines most commonly found are IL10 and TGF-b, which also modulate Treg cell function in vivo and Treg cell development from nonregulatory Materials and methods þ À 14,17,21,22 CD4 CD25 T cells in vitro. However, IL10 Purification and Cultivation of Human CD4 þ T Cells and TGF-b are not essential for the development and 23,24 þ function of Treg cells. CD4 T cells were prepared from peripheral blood CTLA4 also functions as an important mediator of healthy donors by centrifugation over Ficoll- 20 and modulator of Treg cell activity. This member of Hypaque gradients (Biochrom AG, Berlin, Germany) the CD28-family is constitutively expressed at high and MACS isolation using the CD4 þ T cell isolation þ þ levels in CD4 CD25 Treg cells in mouse and man. kit and AutoMACS technology (Miltenyi Biotech, þ þ À In the human system, CTLA4 Treg cells display a Bergisch Gladbach, Germany). CD4 CD25 T cells À þ hi stronger suppressive activity in vitro than CTLA4 and CD4 CD25 Treg cells were then isolated by cell 25 Treg cells. However, CTLA4 is not essential for the sorting on a MoFlo (DakoCytomation, Fort Collins, þ hi development and function of Treg cells, since its loss CO, USA) to a purity of 498%. CD4 CD25 Treg could be compensated by increased secretion of cells were stimulated once with plate-bound anti- À þ þ 26 IL10 and TGF-b by CTLA4 CD4 CD25 Treg cells. CD3 (TR66, 1 mg/ml), soluble anti-CD28 (CD28.2, The conversion of murine naı¨ve nonregulatory 1 mg/ml, BD), and 50 U/ml recombinant human IL2 þ À CD4 CD25 T cells into Treg cells by retroviral (Proleukin, provided by P Wagner, Chiron Corpora- overexpression of FOXP3 underlines the master tion, Emeryville, CA, USA), and thereafter weekly control function of this transcription factor.27,28 with irradiated allogeneic EBV-transformed B cells Consequently, the use of FOXP3 as a tool for (LG2-EBV, provided by T Boon, LICR, Brussels, þ À potential clinical Treg cell therapy has been demon- Belgium). CD4 CD25 T cells were used to establish strated recently in mice. Retroviral transduction of long-term, allo-reactive effector Th cell lines against FOXP3 in polyclonal or antigen-specific naı¨ve LG2-EBV cells. Culture medium was IMDM, with þ À CD4 CD25 T cells generated Treg cells that 10% FCS, 100 U/ml penicillin/streptomycin, and suppressed the development of inflammatory nonessential amino acids (PAA Laboratories, Linz, bowl disease, autoimmune gastritis, diabetes or Austria). Human peripheral blood was obtained contact allergy, and induced transplant tolerance after informed consent according to the MHH in vivo.27,29–31 Despite these encouraging results in guidelines. mice, debate exists about the efficacy of FOXP3 to reprogram human naı¨ve CD4 þ T cells towards a regulatory phenotype.32–34 The most likely reason for Antibodies this ambiguity arises from the fact that unlike For immunostaining PE-, FITC-, APC-, and Cy- murine naı¨ve and effector CD4 þ T helper cells, Chrom-conjugated Abs against CD4 (RPA-T4), human nonregulatory CD4 þ T cells do express CD25 (M-A251), CD28 (CD28.2), TCRab (T10B9.1A- FOXP3.35 31), CTLA4 (BNI3), LGALS3 (B2C10; all from BD, Thus, the fundamental role of FOXP3 is well Bioscience, San Jose, CA, USA), and FOXP3 established, but target genes involved in the control (PCH101, eBioscine Inc., San Diego, CA, USA) and of the FOXP3-dependent regulatory phenotype still the respective isotype controls were used. For require defining.36 However, precise knowledge of FOXP3 staining, murine hybridoma T cell trans- this, including considerations of the differences duced with human FOXP3 or GFP were included as between FOXP3 in mice and men,35 is essential for further control. Anti-CD3e (TR66, produced from the effective manipulation of regulatory T cells as hybridoma supernatants) and anti-CD28 (CD28.2, immunomodulatory therapy. Hence, we compared BD) were used for T-cell stimulation. the genetic program of human CD4 þ CD25hi derived Treg cells, which were expanded in vitro by allo- geneic stimulation and IL2, with that of human allo- Analysing T-Cell Functions reactive effector Th cells, which were reprogrammed by retroviral transduction of FOXP3. Diubiquitin T-cell proliferation and suppressor activity was (UBD) was identified as a gene that is upregulated assessed by stimulating 3 Â 104 T cells in triplicate by FOXP3 overexpression in Th cells similar to Treg with irradiated LG2-EBV B cells or 1 mg/ml plate- cells. UBD overexpression in Th cells demonstrated bound anti-CD3 (TR66) with or without IL2, or 1 mM that it contributes in part to the FOXP3-dependent ionomycin (Sigma, St Louis, MO, USA), in 96-well regulatory phenotype without upregulation of flat-bottom microtiter plates (Nunc, Wiesbaden, FOXP3, and led to the identification of the Germany). For transwell experiments, Th cells were Laboratory Investigation (2006) 86, 724–737 UBD and LGALS3 represent FOXP3-dependent genes F Ocklenburg et al 726 stimulated in 96 flat-bottom plates separated by under the control of an IRES sequence.37 The 0.2 mm-pore transwell inserts (Greiner bio-one, amphotropic PT67 packaging cell line (provided by Frickenhausen, Germany) from the T cells above M Wirth, GBF) was transfected as described else- the transwell.
Load more

Recommended publications
  • Human Lectins, Their Carbohydrate Affinities and Where to Find Them
    biomolecules Review Human Lectins, Their Carbohydrate Affinities and Where to Review HumanFind Them Lectins, Their Carbohydrate Affinities and Where to FindCláudia ThemD. Raposo 1,*, André B. Canelas 2 and M. Teresa Barros 1 1, 2 1 Cláudia D. Raposo * , Andr1 é LAQVB. Canelas‐Requimte,and Department M. Teresa of Chemistry, Barros NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829‐516 Caparica, Portugal; [email protected] 12 GlanbiaLAQV-Requimte,‐AgriChemWhey, Department Lisheen of Chemistry, Mine, Killoran, NOVA Moyne, School E41 of ScienceR622 Co. and Tipperary, Technology, Ireland; canelas‐ [email protected] NOVA de Lisboa, 2829-516 Caparica, Portugal; [email protected] 2* Correspondence:Glanbia-AgriChemWhey, [email protected]; Lisheen Mine, Tel.: Killoran, +351‐212948550 Moyne, E41 R622 Tipperary, Ireland; [email protected] * Correspondence: [email protected]; Tel.: +351-212948550 Abstract: Lectins are a class of proteins responsible for several biological roles such as cell‐cell in‐ Abstract:teractions,Lectins signaling are pathways, a class of and proteins several responsible innate immune for several responses biological against roles pathogens. such as Since cell-cell lec‐ interactions,tins are able signalingto bind to pathways, carbohydrates, and several they can innate be a immuneviable target responses for targeted against drug pathogens. delivery Since sys‐ lectinstems. In are fact, able several to bind lectins to carbohydrates, were approved they by canFood be and a viable Drug targetAdministration for targeted for drugthat purpose. delivery systems.Information In fact, about several specific lectins carbohydrate were approved recognition by Food by andlectin Drug receptors Administration was gathered for that herein, purpose. plus Informationthe specific organs about specific where those carbohydrate lectins can recognition be found by within lectin the receptors human was body.
    [Show full text]
  • CLEC2B Recombinant Protein Description Product Info
    ABGENEX Pvt. Ltd., E-5, Infocity, KIIT Post Office, Tel : +91-674-2720712, +91-9437550560 Email : [email protected] Bhubaneswar, Odisha - 751024, INDIA 32-3546: CLEC2B Recombinant Protein Alternative C-type lectin domain family 2 member B,Activation-induced C-type lectin,C-type lectin superfamily member Name : 2,IFN-alpha-2b-inducing-related protein 1,CLEC2B,AICL,CLECSF2,IFNRG1,HP10085. Description Source : Escherichia Coli. CLEC2B Human Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain containing 147 amino acids (26-149 a.a) and having a molecular mass of 16.9kDa.CLEC2B is fused to a 23 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques. C-type Lectin Domain Family 2, Member B (CLEC2B) belongs to the C-type lectin/C-type lectin-like domain (CTL/CTLD) superfamily. Members of the CTL/CTLD family share a common protein fold and have various functions, such as cell adhesion, cell-cell signaling, glycoprotein turnover, and roles in inflammation and immune response. The CLEC2B gene is closely connected to other CTL/CTLD superfamily members on chromosome 12p13 in the natural killer gene complex region. CLEC2B is expressed preferentially in lymphoid tissues, and in most hematopoietic cell types. Product Info Amount : 20 µg Purification : Greater than 90.0% as determined by SDS-PAGE. CLEC2B protein solution (1mg/ml) containing 20mM Tris-HCl buffer (pH 8.0), 2M UREA and 10% Content : glycerol. Store at 4°C if entire vial will be used within 2-4 weeks. Store, frozen at -20°C for longer periods of Storage condition : time.
    [Show full text]
  • Fibroblasts from the Human Skin Dermo-Hypodermal Junction Are
    cells Article Fibroblasts from the Human Skin Dermo-Hypodermal Junction are Distinct from Dermal Papillary and Reticular Fibroblasts and from Mesenchymal Stem Cells and Exhibit a Specific Molecular Profile Related to Extracellular Matrix Organization and Modeling Valérie Haydont 1,*, Véronique Neiveyans 1, Philippe Perez 1, Élodie Busson 2, 2 1, 3,4,5,6, , Jean-Jacques Lataillade , Daniel Asselineau y and Nicolas O. Fortunel y * 1 Advanced Research, L’Oréal Research and Innovation, 93600 Aulnay-sous-Bois, France; [email protected] (V.N.); [email protected] (P.P.); [email protected] (D.A.) 2 Department of Medical and Surgical Assistance to the Armed Forces, French Forces Biomedical Research Institute (IRBA), 91223 CEDEX Brétigny sur Orge, France; [email protected] (É.B.); [email protected] (J.-J.L.) 3 Laboratoire de Génomique et Radiobiologie de la Kératinopoïèse, Institut de Biologie François Jacob, CEA/DRF/IRCM, 91000 Evry, France 4 INSERM U967, 92260 Fontenay-aux-Roses, France 5 Université Paris-Diderot, 75013 Paris 7, France 6 Université Paris-Saclay, 78140 Paris 11, France * Correspondence: [email protected] (V.H.); [email protected] (N.O.F.); Tel.: +33-1-48-68-96-00 (V.H.); +33-1-60-87-34-92 or +33-1-60-87-34-98 (N.O.F.) These authors contributed equally to the work. y Received: 15 December 2019; Accepted: 24 January 2020; Published: 5 February 2020 Abstract: Human skin dermis contains fibroblast subpopulations in which characterization is crucial due to their roles in extracellular matrix (ECM) biology.
    [Show full text]
  • ADAR1 Promotes Malignant Progenitor Reprogramming in Chronic Myeloid Leukemia
    ADAR1 promotes malignant progenitor reprogramming in chronic myeloid leukemia Qingfei Jianga,b,c, Leslie A. Crewsa,b,c, Christian L. Barrettd, Hye-Jung Chune, Angela C. Courta,b,c, Jane M. Isquitha,b,c,f, Maria A. Zipetoa,b,c,g, Daniel J. Goffa,b,c, Mark Mindenh, Anil Sadarangania,b,c, Jessica M. Rusertc,i, Kim-Hien T. Daoj, Sheldon R. Morrisa,b, Lawrence S. B. Goldsteinc,i,k, Marco A. Marrae, Kelly A. Frazerd, and Catriona H. M. Jamiesona,b,c,1 aStem Cell Program, Department of Medicine, bMoores Cancer Center, dDivision of Genome Information Sciences, Department of Pediatrics, iDepartment of Cellular and Molecular Medicine, and kHoward Hughes Medical Institute, University of California at San Diego, La Jolla, CA 92093; eCanada’s Michael Smith Genome Sciences Centre, BC Cancer Agency, Vancouver, BC, Canada V5Z 1L3; fDepartment of Animal Science, California Polytechnic State University, San Luis Obispo, CA 93405; gDepartment of Health Sciences, University of Milano-Bicocca, 20052 Milan, Italy; hPrincess Margaret Hospital, Toronto, ON, Canada M5T 2M9; jCenter for Hematologic Malignancies, Oregon Health and Science University Knight Cancer Institute, Portland, OR 97239; and cSanford Consortium for Regenerative Medicine, La Jolla, CA 92037 Edited* by Irving L. Weissman, Stanford University, Stanford, CA, and approved November 30, 2012 (received for review July 30, 2012) The molecular etiology of human progenitor reprogramming into ADAR2 are active in embryonic cell types (18), and ADAR3 self-renewing leukemia stem cells (LSC) has remained elusive. Al- may play a nonenzymatic regulatory role in RNA editing activity though DNA sequencing has uncovered spliceosome gene muta- (22).
    [Show full text]
  • Sloma Et Al CML and NA9 Haematologica-Supplementary
    SUPPLEMENTARY APPENDIX Epigenetic and functional changes imposed by NUP98-HOXA9 in a genetically engineered model of chronic myeloid leukemia progression Ivan Sloma, 1-3° Philip Beer, 1 Christophe Desterke, 4 Elizabeth Bulaeva, 1 Misha Bilenky, 5 Annaïck Carles, 6 Michelle Moksa, 6 Kamini Raghu - ram, 1 Cedric Brimacombe, 1 Karen Lambie, 1 Ali G. Turhan, 7 Orianne Wagner-Ballon, 2,3 Philippe Gaulard, 2,8 Keith Humphries, 1 Martin Hirst 5,6 and Connie J. Eaves 1,9 1Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, British Columbia, Canada; 2Université Paris Est Creteil, INSERM, IMRB, Creteil, France; 3Assistance Publique des Hôpitaux de Paris (AP-HP), Hopital Henri Mondor, Departement d’Hematologie et Immunologie, Creteil, France; 4Université Paris-Sud, Faculté de Médecine Kremlin Bicêtre, INSERM UMS 33 Villejuif, France; 5Canada’s Michael Smith Genome Sciences Centre, British Columbia Cancer Agency, Vancouver, British Columbia, Canada; 6Department of Microbiology and Immunology, Michael Smith Laboratories, University of British Colum - bia, Vancouver, British Columbia, Canada; 7Service d’Hématologie Biologique, Hôpital Universitaire Paris-Saclay, INSERM U935, AP-HP, Kremlin Bicêtre, France; 8INSERM UMRS955, Pathology Department, Hôpital Henri Mondor, AP-HP, Faculté de Médecine de Créteil, Université Paris-Est Créteil, Créteil, France and 9University of British Columbia, Department of Medical Genetics, Vancouver, British Columbia, Canada °Present address: Université Paris Est Creteil, INSERM, IMRB and AP-HP, Hopital Henri Mondor, Departement d’Hematologie et Immunologie, Creteil, France Correspondence: IVAN SLOMA - [email protected] doi:10.3324/haematol.2020.249243 Epigenetic and functional changes imposed by NUP98-HOXA9 in a genetically engineered model of chronic myeloid leukemia progression SUPPLEMENTARY INFORMATION SUPPLEMENTARY MATERIALS AND METHODS Primary cells Heparin-treated blood or leukapheresis cells were obtained from 3 CP CML patients with elevated WBC counts (Supplementary table 1).
    [Show full text]
  • Detecting Methylation Signatures in Neurodegenerative Disease by Density‑Based Clustering of Applications with Reducing Noise Saurav Mallik1 & Zhongming Zhao1,2,3*
    www.nature.com/scientificreports OPEN Detecting methylation signatures in neurodegenerative disease by density‑based clustering of applications with reducing noise Saurav Mallik1 & Zhongming Zhao1,2,3* There have been numerous genetic and epigenetic datasets generated for the study of complex disease including neurodegenerative disease. However, analysis of such data often sufers from detecting the outliers of the samples, which subsequently afects the extraction of the true biological signals involved in the disease. To address this critical issue, we developed a novel framework for identifying methylation signatures using consecutive adaptation of a well‑known outlier detection algorithm, density based clustering of applications with reducing noise (DBSCAN) followed by hierarchical clustering. We applied the framework to two representative neurodegenerative diseases, Alzheimer’s disease (AD) and Down syndrome (DS), using DNA methylation datasets from public sources (Gene Expression Omnibus, GEO accession ID: GSE74486). We frst applied DBSCAN algorithm to eliminate outliers, and then used Limma statistical method to determine diferentially methylated genes. Next, hierarchical clustering technique was applied to detect gene modules. Our analysis identifed a methylation signature comprising 21 genes for AD and a methylation signature comprising 89 genes for DS, respectively. Our evaluation indicated that these two signatures could lead to high classifcation accuracy values (92% and 70%) for these two diseases. In summary, this framework will be useful to better detect outlier‑free genetic and epigenetic signatures in various complex diseases and their developmental stages. Te past 2 decades have witnessed exponential growth of genetic and epigenetic data generation, which sub- stantially helps the advancement of biological and biomedical research.
    [Show full text]
  • Analysis of Tissue-Specific & Allele- Specific DNA Methylation
    Analysis of tissue-specific & allele- specific DNA methylation Dissertation zur Erlangung des Doktorgrades der Naturwissenschaften (Dr. rer. nat.) der Naturwissenschaftlichen Fakultät IV – Chemie und Pharmazie der Universität Regensburg vorgelegt von Elmar Schilling aus Schwenningen 2009 The present work was carried out in the Department of Hematology and Oncology at the University Hospital Regensburg from June 2005 to June 2009 and was supervised by PD. Dr. Michael Rehli. Die vorliegende Arbeit entstand in der Zeit von Juni 2005 bis Juni 2009 in der Abteilung für Hämatologie und internistische Onkologie des Klinikums der Universität Regensburg unter der Anleitung von PD. Dr. Michael Rehli. Promotionsgesuch eingereicht am: 30. Juli 2009 Die Arbeit wurde angeleitet von PD. Dr. Michael Rehli. Prüfungsausschuss: Vorsitzender: Prof. Dr. Sigurd Elz 1. Gutachter: Prof. Dr. Roland Seifert 2. Gutachter: PD. Dr. Michael Rehli 3. Prüfer: Prof. Dr. Gernot Längst Lob und Tadel bringen den Weisen nicht aus dem Gleichgewicht. (Budha) TABLE OF CONTENTS 1 INTRODUCTION .............................................................................................. 1 1.1 THE CONCEPT OF EPIGENETICS ............................................................................................. 1 1.2 DNA METHYLATION .............................................................................................................. 2 1.2.1 DNA methyltransferases ................................................................................................ 3 1.2.2
    [Show full text]
  • Antibody Repertoire and Gene Expression Dynamics of Diverse Human B Cell
    bioRxiv preprint doi: https://doi.org/10.1101/2020.04.28.054775; this version posted May 26, 2020. 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 4.0 International license. 1 Antibody repertoire and gene expression dynamics of diverse human B cell 2 states during affinity maturation. 3 Hamish W King1,2 *, Nara Orban3, John C Riches4,5, Andrew J Clear4, Gary Warnes6, Sarah A 4 Teichmann2,7, Louisa K James1 * § 5 1 Centre for Immunobiology, Blizard Institute, Queen Mary University of London, London E1 2AT, UK 6 2 Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge CB10 1SA, UK 7 3 Barts Health Ear, Nose and Throat Service, The Royal London Hospital, London E1 1BB, UK 8 4 Centre for Haemato-Oncology, Barts Cancer Institute, Queen Mary University of London, London EC1M 6BQ, UK 9 5 The Francis Crick Institute, London NW1 1AT, UK 10 6 Flow Cytometry Core Facility, Blizard Institute, Queen Mary University of London, London E1 2AT, UK 11 7 Theory of Condensed Matter, Cavendish Laboratory, Department of Physics, University of Cambridge, Cambridge CB3 0EH, UK 12 § Lead contact 13 * To whom correspondence can be addressed: [email protected], [email protected] and [email protected] 14 15 Abstract 16 In response to antigen challenge, B cells clonally expand, undergo selection and differentiate to produce 17 mature B cell subsets and high affinity antibodies.
    [Show full text]
  • Interaction of C-Type Lectin-Like Receptors Nkp65 and KACL Facilitates Dedicated Immune Recognition of Human Keratinocytes
    Interaction of C-type lectin-like receptors NKp65 and KACL facilitates dedicated immune recognition of human keratinocytes Jessica Spreua, Sabrina Kuttruffa, Veronika Stejfovaa, Kevin M. Dennehya, Birgit Schittekb, and Alexander Steinlea,c,1 aDepartment of Immunology, Institute for Cell Biology, and bDepartment of Dermatology, Eberhard-Karls-University Tübingen, 72076 Tübingen, Germany; and cInstitute for Molecular Medicine, Goethe-University Frankfurt, 60590 Frankfurt, Germany Edited* by Wayne M. Yokoyama, Washington University School of Medicine, St. Louis, MO, and approved February 3, 2010 (received for review November 15, 2009) Many well-known immune-related C-type lectin-like receptors remain unknown, the immunobiology of these NKC-based recep- (CTLRs) such as NKG2D, CD69, and the Ly49 receptors are encoded tor-ligand systems is far from being understood. Also in humans, in the natural killer gene complex (NKC). Recently, we character- corresponding NKRP1–CLEC2 receptor–ligand pairs have ized the orphan NKC gene CLEC2A encoding for KACL, a further recently been characterized: LLT1 (encoded by CLEC2D)was member of the human CLEC2 family of CTLRs. In contrast to the identified as a ligand of the inhibitory NK receptor NKR-P1A/ other CLEC2 family members AICL, CD69, and LLT1, KACL expres- CD161 (11, 12) and another member of the human CLEC2 family, sion is mostly restricted to skin. Here we show that KACL is a non– AICL (CLEC2B), as a ligand of the activating NK receptor NKp80 disulfide-linked homodimeric surface receptor and stimulates cyto- (13). LLT1 is expressed by activated DC and B cells and was shown toxicity by human NK92MI cells. We identified the corresponding to inhibit NK cytotoxicity, whereas AICL on myeloid cells stim- activating receptor on NK92MI cells that is encoded adjacently to ulates NK cytotoxicity as well as a cross-talk between NK cells and the CLEC2A locus and binds KACL with high affinity.
    [Show full text]
  • Supplementary Table 1 Double Treatment Vs Single Treatment
    Supplementary table 1 Double treatment vs single treatment Probe ID Symbol Gene name P value Fold change TC0500007292.hg.1 NIM1K NIM1 serine/threonine protein kinase 1.05E-04 5.02 HTA2-neg-47424007_st NA NA 3.44E-03 4.11 HTA2-pos-3475282_st NA NA 3.30E-03 3.24 TC0X00007013.hg.1 MPC1L mitochondrial pyruvate carrier 1-like 5.22E-03 3.21 TC0200010447.hg.1 CASP8 caspase 8, apoptosis-related cysteine peptidase 3.54E-03 2.46 TC0400008390.hg.1 LRIT3 leucine-rich repeat, immunoglobulin-like and transmembrane domains 3 1.86E-03 2.41 TC1700011905.hg.1 DNAH17 dynein, axonemal, heavy chain 17 1.81E-04 2.40 TC0600012064.hg.1 GCM1 glial cells missing homolog 1 (Drosophila) 2.81E-03 2.39 TC0100015789.hg.1 POGZ Transcript Identified by AceView, Entrez Gene ID(s) 23126 3.64E-04 2.38 TC1300010039.hg.1 NEK5 NIMA-related kinase 5 3.39E-03 2.36 TC0900008222.hg.1 STX17 syntaxin 17 1.08E-03 2.29 TC1700012355.hg.1 KRBA2 KRAB-A domain containing 2 5.98E-03 2.28 HTA2-neg-47424044_st NA NA 5.94E-03 2.24 HTA2-neg-47424360_st NA NA 2.12E-03 2.22 TC0800010802.hg.1 C8orf89 chromosome 8 open reading frame 89 6.51E-04 2.20 TC1500010745.hg.1 POLR2M polymerase (RNA) II (DNA directed) polypeptide M 5.19E-03 2.20 TC1500007409.hg.1 GCNT3 glucosaminyl (N-acetyl) transferase 3, mucin type 6.48E-03 2.17 TC2200007132.hg.1 RFPL3 ret finger protein-like 3 5.91E-05 2.17 HTA2-neg-47424024_st NA NA 2.45E-03 2.16 TC0200010474.hg.1 KIAA2012 KIAA2012 5.20E-03 2.16 TC1100007216.hg.1 PRRG4 proline rich Gla (G-carboxyglutamic acid) 4 (transmembrane) 7.43E-03 2.15 TC0400012977.hg.1 SH3D19
    [Show full text]
  • Risk Variants Disrupting Enhancers of TH1 and TREG Cells in Type 1 Diabetes
    Risk variants disrupting enhancers of TH1 and TREG cells in type 1 diabetes Peng Gaoa,b,c,1, Yasin Uzuna,b,c,1, Bing Hea,b,c, Sarah E. Salamatid, Julie K. M. Coffeyd, Eva Tsalikiand, and Kai Tana,b,c,e,f,g,2 aDivision of Oncology, Children’s Hospital of Philadelphia, Philadelphia, PA 19104; bCenter for Childhood Cancer Research, Children’s Hospital of Philadelphia, Philadelphia, PA 19104; cDepartment of Biomedical and Health Informatics, Children’s Hospital of Philadelphia, Philadelphia, PA 19104; dStead Family Department of Pediatrics, University of Iowa Carver College of Medicine, Iowa City, IA 52242; eDepartment of Pediatrics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104; fDepartment of Genetics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104; and gDepartment of Cell and Developmental Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104 Edited by Wing Hung Wong, Stanford University, Stanford, CA, and approved March 1, 2019 (received for review September 5, 2018) Genome-wide association studies (GWASs) have revealed 59 geno- patients, despite of the pivotal roles of TH1 and TREG cells in the mic loci associated with type 1 diabetes (T1D). Functional inter- pathogenesis of T1D. In this study, we conducted epigenomic pretation of the SNPs located in the noncoding region of these loci and transcriptomic profiling of TH1 and TREG cells isolated from remains challenging. We perform epigenomic profiling of two a cohort of five healthy donors and six newly diagnosed T1D enhancer marks, H3K4me1 and H3K27ac, using primary TH1 and patients. Our data (8) reveal significant alteration in the en- TREG cells isolated from healthy and T1D subjects.
    [Show full text]
  • Detection of H3k4me3 Identifies Neurohiv Signatures, Genomic
    viruses Article Detection of H3K4me3 Identifies NeuroHIV Signatures, Genomic Effects of Methamphetamine and Addiction Pathways in Postmortem HIV+ Brain Specimens that Are Not Amenable to Transcriptome Analysis Liana Basova 1, Alexander Lindsey 1, Anne Marie McGovern 1, Ronald J. Ellis 2 and Maria Cecilia Garibaldi Marcondes 1,* 1 San Diego Biomedical Research Institute, San Diego, CA 92121, USA; [email protected] (L.B.); [email protected] (A.L.); [email protected] (A.M.M.) 2 Departments of Neurosciences and Psychiatry, University of California San Diego, San Diego, CA 92103, USA; [email protected] * Correspondence: [email protected] Abstract: Human postmortem specimens are extremely valuable resources for investigating trans- lational hypotheses. Tissue repositories collect clinically assessed specimens from people with and without HIV, including age, viral load, treatments, substance use patterns and cognitive functions. One challenge is the limited number of specimens suitable for transcriptional studies, mainly due to poor RNA quality resulting from long postmortem intervals. We hypothesized that epigenomic Citation: Basova, L.; Lindsey, A.; signatures would be more stable than RNA for assessing global changes associated with outcomes McGovern, A.M.; Ellis, R.J.; of interest. We found that H3K27Ac or RNA Polymerase (Pol) were not consistently detected by Marcondes, M.C.G. Detection of H3K4me3 Identifies NeuroHIV Chromatin Immunoprecipitation (ChIP), while the enhancer H3K4me3 histone modification was Signatures, Genomic Effects of abundant and stable up to the 72 h postmortem. We tested our ability to use H3K4me3 in human Methamphetamine and Addiction prefrontal cortex from HIV+ individuals meeting criteria for methamphetamine use disorder or not Pathways in Postmortem HIV+ Brain (Meth +/−) which exhibited poor RNA quality and were not suitable for transcriptional profiling.
    [Show full text]