DOCSLIB.ORG

978.Full.Pdf

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
978.Full.Pdf Dendritic Cell Migration and Antigen Presentation Are Coordinated by the Opposing Functions of the Tetraspanins CD82 and CD37 This information is current as of September 26, 2021. Eleanor L. Jones, Janet L. Wee, Maria C. Demaria, Jessica Blakeley, Po Ki Ho, Javier Vega-Ramos, Jose A. Villadangos, Annemiek B. van Spriel, Michael J. Hickey, Günther J. Hämmerling and Mark D. Wright J Immunol 2016; 196:978-987; Prepublished online 4 Downloaded from January 2016; doi: 10.4049/jimmunol.1500357 http://www.jimmunol.org/content/196/3/978 http://www.jimmunol.org/ Supplementary http://www.jimmunol.org/content/suppl/2016/01/02/jimmunol.150035 Material 7.DCSupplemental References This article cites 45 articles, 23 of which you can access for free at: http://www.jimmunol.org/content/196/3/978.full#ref-list-1 by guest on September 26, 2021 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 *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. The Journal of Immunology Dendritic Cell Migration and Antigen Presentation Are Coordinated by the Opposing Functions of the Tetraspanins CD82 and CD37 Eleanor L. Jones,* Janet L. Wee,*,† Maria C. Demaria,* Jessica Blakeley,* Po Ki Ho,* Javier Vega-Ramos,‡ Jose A. Villadangos,‡,x Annemiek B. van Spriel,{ Michael J. Hickey,† Gunther€ J. Ha¨mmerling,‖ and Mark D. Wright* This study supports a new concept where the opposing functions of the tetraspanins CD37 and CD82 may coordinate changes in migration and Ag presentation during dendritic cell (DC) activation. We have previously published that CD37 is downregulated upon monocyte-derived DC activation, promotes migration of both skin and bone marrow–derived dendritic cells (BMDCs), and restrains Ag presentation in splenic and BMDCs. In this article, we show that CD82, the closest phylogenetic relative to CD37, Downloaded from appears to have opposing functions. CD82 is upregulated upon activation of BMDCs and monocyte-derived DCs, restrains migration of skin and BMDCs, supports MHC class II maturation, and promotes stable interactions between T cells and splenic DCs or BMDCs. The underlying mechanism involves the rearrangement of the cytoskeleton via a differential activation of small GTPases. Both CD372/2 and CD822/2 BMDCs lack cellular projections, but where CD372/2 BMDCs spread poorly on fibro- nectin, CD822/2 BMDCs are large and spread to a greater extent than wild-type BMDCs. At the molecular level, CD82 is a negative regulator of RhoA, whereas CD37 promotes activation of Rac-1; both tetraspanins negatively regulate Cdc42. Thus, this http://www.jimmunol.org/ study identifies a key aspect of DC biology: an unactivated BMDC is CD37hiCD82lo, resulting in a highly motile cell with a limited ability to activate naive T cells. By contrast, a late activated BMDC is CD37loCD82hi, and thus has modified its migratory, cytoskeletal, and Ag presentation machinery to become a cell superbly adapted to activating naive T cells. The Journal of Immunology, 2016, 196: 978–987. endritic cells (DC) are the most potent of the APCs through pattern recognition receptors, they reduce MHC turnover because they have the unique ability to activate naive Ag- and upregulate expression of MHC/peptide complexes, costimu- D specific T cells (1). However, DC function varies with latory molecules such as CD80 and CD86, and proinflammatory by guest on September 26, 2021 activation state; the classical example being migratory DCs. cytokines, all of which increase their capacity to stimulate T cells Unactivated migratory DCs in the periphery efficiently patrol and direct adaptive cellular immunity (2). Concurrently, DCs through tissues and are specialized for Ag uptake. These highly undergo morphological changes involving the extension of den- endocytic cells are poor stimulators of T cells, because of both drites thought to promote efficient interactions with T cells, a a moderate surface expression and a high turnover of MHC decrease in Ag uptake and processing, and a modification in cell molecules. However, once DCs receive danger signals transduced migration. Thus, rather than migrating randomly through tissue, they now migrate directionally, via the lymphatics to the draining *Department of Immunology and Pathology, Monash University, Melbourne, Victoria lymph nodes (2–4). The mechanisms of migration used by DCs 3004, Australia; †Department of Medicine, Centre for Inflammatory Diseases, Monash have been reported to be both dependent on adhesion molecules University, Clayton, Victoria 3168, Australia; ‡Department of Microbiology and Immu- x (5–7) and adhesion-independent ameboid migration, driven chiefly nology, University of Melbourne, Melbourne 3010, Australia; Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University by cytoskeletal protrusion and contractile forces (8). Nonetheless, of Melbourne, Melbourne 3010, Australia; {Radboud Institute for Molecular Life the activated DC that has migrated to the draining lymph node has Sciences, Radboud University Medical Center, G525 GA Nijmegen, the Netherlands; and ‖ now metamorphosed to become a cell specialized at initiating Molecular Immunology, German Cancer Research Center, 69120 Heidelberg, Germany adaptive immunity by presenting Ags to naive T cells and inducing ORCIDs: 0000-0001-6015-5651 (M.C.D.); 0000-0001-6771-8891 (J.A.V.); 0000- 0002-3590-2368 (A.B.v.S.); 0000-0003-2354-357X (M.J.H.); 0000-0002-2177-5214 their activation (2). These major functional changes require ex- (M.D.W.). quisite coordination between the Ag processing and presentation Received for publication February 12, 2015. Accepted for publication December 1, machinery, and the myriad of adhesion, signaling, and cytoskeletal 2015. proteins that regulate DC morphology and migration. This work was supported by grants from the Australian National Health and Medical One type of protein known to regulate both Ag presentation and Research Council and the Netherlands Organization for Scientific Research (NWO- Vidi Grant 864.11.006 to A.B.v.S.). cell motility is the tetraspanins, a superfamily of four-transmembrane molecules, highly conserved in evolution and expressed in all mam- Address correspondence and reprint requests to Dr. Mark D. Wright, Monash University, Alfred Medical Research and Education Precinct, Commercial Road, malian cells. The best-defined role of tetraspanins in biology is their Melbourne, VIC 4004, Australia. E-mail address: [email protected] molecular organization of cell membranes. Tetraspanins directly The online version of this article contains supplemental material. interact with their molecular partners and organize them into signal- Abbreviations used in this article: BMDC, bone marrow–derived DC; DC, dendritic transducing microdomains (9, 10). Components of tetraspanin- cell; MHC II, MHC class II; MoDC, monocyte-derived DC; TEM, tetraspanin-enriched enriched microdomains (TEM) include membrane proteins such as microdomain; WT, wild-type. integrins, proteases that regulate cell-surface molecule expression Copyright Ó 2016 by The American Association of Immunologists, Inc. 0022-1767/16/$30.00 posttranslationally, and signaling molecules including kinases and www.jimmunol.org/cgi/doi/10.4049/jimmunol.1500357 The Journal of Immunology 979 phosphatases. Tetraspanins regulate the spatiotemporal molecular following primers: forward: 59-AGGTGTTTGCCCTTCTCCTT-39 and 2/2 interactions of their partner proteins and thereby influence complex reverse: 59-CCACCTGTGACAACCAAGTG-39. CD82 PCR was per- cellular events such as activation, adhesion, and migration. An formed with the following primers: forward: 59-TCCTTAAGCCTCAA- GAAAACC-39 and reverse: 59-TGTGAGGGCTCCAGTCTCC-39. PCR emerging role for TEMs is in communication between the cell program was as follows: 95˚C for 15 min, cycle: 95˚C 30 s, 50˚C 30 s, surface and the cytoskeleton. Tetraspanins regulate cytoskeletal- 72˚C 45 s, 35 cycles, 72˚C for 7 min (final annealing), and hold at 4˚C. 2 2 dependent processes such as outside-in integrin signaling and CD82 / mice were shown to lack CD82 mRNA, and levels of CD82 in WT BMDCs were measured via TaqMan real-time PCR for mouse CD82 adhesion strengthening, actin polymerization, cellular polarity, 2 2 (CD82 / expression of CD82 not shown) (Catalog Mm00492061_m1; and spreading (9–11). On the molecular level the tetraspanin Life Technologies). Levels of CD37 in WT BMDCs were measured via CD81 has been shown to interact with three important regulators TaqMan real-time PCR for mouse CD37 (Catalog Mm00514240_m1; Life of cytoskeleton polymerization: the Rho GTPase Rac, Ezrin, and Technologies). Levels of 18S were measured by TaqMan real-time PCR Moesin. Ezrin and Moesin are peripheral membrane proteins that for mouse 18S (Catalog Mm_03928990_g1; Life Technologies). Real-time
Load more

Recommended publications
  • Human and Mouse CD Marker Handbook Human and Mouse CD Marker Key Markers - Human Key Markers - Mouse
    Welcome to More Choice CD Marker Handbook For more information, please visit: Human bdbiosciences.com/eu/go/humancdmarkers Mouse bdbiosciences.com/eu/go/mousecdmarkers Human and Mouse CD Marker Handbook Human and Mouse CD Marker Key Markers - Human Key Markers - Mouse CD3 CD3 CD (cluster of differentiation) molecules are cell surface markers T Cell CD4 CD4 useful for the identification and characterization of leukocytes. The CD CD8 CD8 nomenclature was developed and is maintained through the HLDA (Human Leukocyte Differentiation Antigens) workshop started in 1982. CD45R/B220 CD19 CD19 The goal is to provide standardization of monoclonal antibodies to B Cell CD20 CD22 (B cell activation marker) human antigens across laboratories. To characterize or “workshop” the antibodies, multiple laboratories carry out blind analyses of antibodies. These results independently validate antibody specificity. CD11c CD11c Dendritic Cell CD123 CD123 While the CD nomenclature has been developed for use with human antigens, it is applied to corresponding mouse antigens as well as antigens from other species. However, the mouse and other species NK Cell CD56 CD335 (NKp46) antibodies are not tested by HLDA. Human CD markers were reviewed by the HLDA. New CD markers Stem Cell/ CD34 CD34 were established at the HLDA9 meeting held in Barcelona in 2010. For Precursor hematopoetic stem cell only hematopoetic stem cell only additional information and CD markers please visit www.hcdm.org. Macrophage/ CD14 CD11b/ Mac-1 Monocyte CD33 Ly-71 (F4/80) CD66b Granulocyte CD66b Gr-1/Ly6G Ly6C CD41 CD41 CD61 (Integrin b3) CD61 Platelet CD9 CD62 CD62P (activated platelets) CD235a CD235a Erythrocyte Ter-119 CD146 MECA-32 CD106 CD146 Endothelial Cell CD31 CD62E (activated endothelial cells) Epithelial Cell CD236 CD326 (EPCAM1) For Research Use Only.
    [Show full text]
  • B-Cell Receptor Pathway Inhibitors Affect CD20 Levels and Impair Antitumor Activity of Anti-CD20 Monoclonal Antibodies
    Letters to the Editor 1163 13 Kuruvilla J, Gutierrez M, Shah BD, Gabrail NY, de Nully Brown P, 14 Yu L, Mohamed AJ, Simonson OE, Vargas L, Blomberg KE, Bjorkstrand B et al. Stone RM et al. Preliminary evidence of anti tumor activity of selinexor Proteasome-dependent autoregulation of Bruton tyrosine kinase (Btk) promoter (KPT-330) in a phase I trial of a first-in-class oral selective inhibitor via NF-kappaB. Blood 2008; 111: 4617–4626. of nuclear export (SINE) in patients (pts) with relapsed/refractory non 15BurgerJA,BurgerM,KippsTJ.Chronic lymphocytic leukemia B cells Hodgkin’s lymphoma (NHL) and chronic lymphocytic leukemia (CLL). Blood 2013; express functional CXCR4 chemokine receptors that mediate spontaneous 122: 90. migration beneath bone marrow stromal cells. Blood 1999; 94: 3658–3667. Supplementary Information accompanies this paper on the Leukemia website (http://www.nature.com/leu) B-cell receptor pathway inhibitors affect CD20 levels and impair antitumor activity of anti-CD20 monoclonal antibodies Leukemia (2014) 28, 1163–1167; doi:10.1038/leu.2014.12 also tested a primary MCL sample and upon treatment with BCR inhibitors observed a significant downregulation of surface CD20 levels and a trend towards impaired R-CDC and O-CDC (Supplementary Figure 1b). Moreover, we determined the Signaling via the aberrantly activated B-cell receptor (BCR) has a influence of BCR inhibitors on CD20 surface levels in a critical role in the pathogenesis of B-cell tumors by promoting series of 15 tumor cell lines, including Burkitt’s lymphoma (Ramos, survival and clonal expansion of malignant B cells.1,2 Multiple Daudi and BJAB), ALL (NALM-6), diffuse large B-cell lymphoma preclinical studies indicate that blocking various components of (BCR-dependent Ly-1, Ly-7, Ly-10, DHL-6, HBL-1, U2932 and the BCR signaling pathway holds a great therapeutic potential in BCR-independent Ly-4, Ly-19, Pfeiffer) and CLL (EHEB and MEC-1).
    [Show full text]
  • Dynamics Within Tetraspanin Pairs Affect MHC Class II Expression
    328 Research Article Dynamics within tetraspanin pairs affect MHC class II expression Tineke van den Hoorn, Petra Paul, Lennert Janssen, Hans Janssen and Jacques Neefjes* Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands *Author for correspondence ([email protected]) Accepted 11 August 2011 Journal of Cell Science 125, 328–339 ß 2012. Published by The Company of Biologists Ltd doi: 10.1242/jcs.088047 Summary Late endosomal multivesicular bodies (MVBs) are complicated organelles with various subdomains located at the limiting membrane and the internal vesicles (ILVs). ILVs accumulate tetraspanins such as CD63 and CD82 that might form protein assemblies, including major histocompatibility complex class II (MHC-II) and its chaperone human leukocyte antigen (HLA)-DM. Here, we studied the effect of four late endosomal tetraspanin proteins on MHC-II expression. Silencing CD9, CD63 and CD81 enhanced MHC-II expression whereas silencing CD82 did not. No effect on peptide loading was observed. Using confocal FRET technology, we measured the dynamics of CD63 and CD82 interaction with MHC-II and its chaperone HLA-DM. CD63–CD82 interactions remained unaltered in the two MVB subdomains whereas the interactions between CD63 or CD82 homologous pairs changed. CD63 stably associated with MHC- II, and CD82 with HLA-DM, on both MVB subdomains whereas the CD82–MHC-II and CD63–HLA-DM interactions changed. These data visualize for the first time the protein dynamics of tetraspanin assemblies in MVB
    [Show full text]
  • Altered Expression of CD63 and Exosomes in Scleroderma Dermal
    Journal of Dermatological Science 84 (2016) 30–39 Contents lists available at ScienceDirect Journal of Dermatological Science journal homepage: www.jdsjournal.com Altered expression of CD63 and exosomes in scleroderma dermal fibroblasts Kayo Nakamura, Masatoshi Jinnin*, Miho Harada, Hideo Kudo, Wakana Nakayama, Kuniko Inoue, Aki Ogata, Ikko Kajihara, Satoshi Fukushima, Hironobu Ihn Department of Dermatology and Plastic Surgery, Faculty of Life Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-8556, Japan A R T I C L E I N F O A B S T R A C T Article history: Background: Exosomes are small vesicles shed from various cells. They contain proteins, lipids, and Received 6 January 2016 nucleic acids, and are regarded as a tool of cell-cell communication. Received in revised form 13 June 2016 Objectives: To reveal the putative role of exosomes in systemic sclerosis (SSc), and to elucidate the effect of Accepted 29 June 2016 exosomes on wound healing. Methods: The expression of common markers for exosomes (CD63, CD9, and CD81) and type I collagen Keywords: were examined with real-time PCR, immunohistochemical analysis, ELISA, immunoblotting, and flow Exosomes cytometry. The effect of serum-derived exosomes on wound healing was tested on full-thickness wounds CD63 in the mid-dorsal skin of BALB/c mice. Systemic sclerosis Results: The expression levels of CD63 as well as CD9 and CD81 tended to be increased in SSc dermal fibroblasts compared to normal fibroblasts. Increased exosomes in a cultured media of SSc fibroblasts stimulated the expression levels of type I collagen in normal fibroblasts. As the mechanism, collagen- related microRNA levels in SSc fibroblast-derived exosomes were dysregulated, indicating that both the amount and the content of exosomes were altered in SSc.
    [Show full text]
  • Combination Immunotherapy with Anti-CD20 and Anti-HLA-DR Monoclonal Antibodies Induces Synergistic Anti-Lymphoma Effects in Human Lymphoma Cell Lines
    UC Davis UC Davis Previously Published Works Title Combination immunotherapy with anti-CD20 and anti-HLA-DR monoclonal antibodies induces synergistic anti-lymphoma effects in human lymphoma cell lines Permalink https://escholarship.org/uc/item/8pk1f4nx Journal Leukemia & Lymphoma, 48(5) ISSN 1042-8194 Authors Tobin, Evan Denardo, Gerald Zhang, Nan et al. Publication Date 2007-05-01 Peer reviewed eScholarship.org Powered by the California Digital Library University of California Rituximab & ChLym-1 Combined Immunotherapy Combination Immunotherapy with Anti-CD20 and Anti-HLA-DR Monoclonal Antibodies Induces Synergistic Anti-lymphoma Effects in Human Lymphoma Cell Lines Evan Tobin1, Gerald DeNardo1, Nan Zhang2, Alan L. Epstein2, Cathy Liu1 & Sally DeNardo1 1 Department of Internal Medicine, University of California Davis, CA, USA 2 Department of Pathology, University of Southern California Keck School of Medicine, Los Angeles, CA, USA Running Title: Rituximab & ChLym-1 Combined Immunotherapy Keywords: Lymphoma; immunotherapy; rituximab; Lym-1; CD20; HLA-DR 1Address for correspondence: Gerald L. DeNardo, M.D. Division of Hematology and Oncology 1508 Alhambra Blvd., No. 3100 Sacramento, CA 95816 Telephone (916) 734-3787 Fax (916) 703-5014 E-mail: [email protected] 1 Rituximab & ChLym-1 Combined Immunotherapy ABSTRACT Rituximab is effective in about one half of patients with indolent lymphoma. Even these patients relapse and develop rituximab resistance. To increase potency and circumvent resistance, the anti-lymphoma effects of rituximab, an anti-CD20 MAb1, combined with chLym-12, an anti- HLA-DR MAb, were assessed in human lymphoma cell lines by examining growth inhibition and cell death, apoptosis induction, ADCC3 and CDC4. There were additive effects in all assays and synergism in cell lines, such as B35M, which displayed resistance to either MAb alone.
    [Show full text]
  • Pancancer Progression Human Vjune2017
    Gene Symbol Accession Alias/Prev Symbol Official Full Name AAMP NM_001087.3 - angio-associated, migratory cell protein ABI3BP NM_015429.3 NESHBP|TARSH ABI family, member 3 (NESH) binding protein ACHE NM_000665.3 ACEE|ARACHE|N-ACHE|YT acetylcholinesterase ACTG2 NM_001615.3 ACT|ACTA3|ACTE|ACTL3|ACTSG actin, gamma 2, smooth muscle, enteric ACVR1 NM_001105.2 ACTRI|ACVR1A|ACVRLK2|ALK2|FOP|SKR1|TSRI activin A receptor, type I ACVR1C NM_145259.2 ACVRLK7|ALK7 activin A receptor, type IC ACVRL1 NM_000020.1 ACVRLK1|ALK-1|ALK1|HHT|HHT2|ORW2|SKR3|TSR-I activin A receptor type II-like 1 ADAM15 NM_207195.1 MDC15 ADAM metallopeptidase domain 15 ADAM17 NM_003183.4 ADAM18|CD156B|CSVP|NISBD|TACE ADAM metallopeptidase domain 17 ADAM28 NM_014265.4 ADAM 28|ADAM23|MDC-L|MDC-Lm|MDC-Ls|MDCL|eMDC II|eMDCII ADAM metallopeptidase domain 28 ADAM8 NM_001109.4 CD156|MS2 ADAM metallopeptidase domain 8 ADAM9 NM_001005845.1 CORD9|MCMP|MDC9|Mltng ADAM metallopeptidase domain 9 ADAMTS1 NM_006988.3 C3-C5|METH1 ADAM metallopeptidase with thrombospondin type 1 motif, 1 ADAMTS12 NM_030955.2 PRO4389 ADAM metallopeptidase with thrombospondin type 1 motif, 12 ADAMTS8 NM_007037.4 ADAM-TS8|METH2 ADAM metallopeptidase with thrombospondin type 1 motif, 8 ADAP1 NM_006869.2 CENTA1|GCS1L|p42IP4 ArfGAP with dual PH domains 1 ADD1 NM_001119.4 ADDA adducin 1 (alpha) ADM2 NM_001253845.1 AM2|dJ579N16.4 adrenomedullin 2 ADRA2B NM_000682.4 ADRA2L1|ADRA2RL1|ADRARL1|ALPHA2BAR|alpha-2BAR adrenoceptor alpha 2B AEBP1 NM_001129.3 ACLP AE binding protein 1 AGGF1 NM_018046.3 GPATC7|GPATCH7|HSU84971|HUS84971|VG5Q
    [Show full text]
  • Aberrant Expression of Tetraspanin Molecules in B-Cell Chronic Lymphoproliferative Disorders and Its Correlation with Normal B-Cell Maturation
    Leukemia (2005) 19, 1376–1383 & 2005 Nature Publishing Group All rights reserved 0887-6924/05 $30.00 www.nature.com/leu Aberrant expression of tetraspanin molecules in B-cell chronic lymphoproliferative disorders and its correlation with normal B-cell maturation S Barrena1,2, J Almeida1,2, M Yunta1,ALo´pez1,2, N Ferna´ndez-Mosteirı´n3, M Giralt3, M Romero4, L Perdiguer5, M Delgado1, A Orfao1,2 and PA Lazo1 1Instituto de Biologı´a Molecular y Celular del Ca´ncer, Centro de Investigacio´n del Ca´ncer, Consejo Superior de Investigaciones Cientı´ficas-Universidad de Salamanca, Salamanca, Spain; 2Servicio de Citometrı´a, Universidad de Salamanca and Hospital Universitario de Salamanca, Salamanca, Spain; 3Servicio de Hematologı´a, Hospital Universitario Miguel Servet, Zaragoza, Spain; 4Hematologı´a-hemoterapia, Hospital Universitario Rı´o Hortega, Valladolid, Spain; and 5Servicio de Hematologı´a, Hospital de Alcan˜iz, Teruel, Spain Tetraspanin proteins form signaling complexes between them On the cell surface, tetraspanin antigens are present either as and with other membrane proteins and modulate cell adhesion free molecules or through interaction with other proteins.25,26 and migration properties. The surface expression of several tetraspanin antigens (CD9, CD37, CD53, CD63, and CD81), and These interacting proteins include other tetraspanins, integri- F 22,27–30F their interacting proteins (CD19, CD21, and HLA-DR) were ns particularly those with the b1 subunit HLA class II 31–33 34,35 analyzed during normal B-cell maturation and compared to a moleculesFeg HLA DR -, CD19, the T-cell recep- group of 67 B-cell neoplasias. Three patterns of tetraspanin tor36,37 and several other members of the immunoglobulin expression were identified in normal B cells.
    [Show full text]
  • KAI Expression Prevents IL-8-Mediated Endothelial Gap Formation in Late-Stage Melanomas
    Oncogene (2014) 33, 2898–2908 & 2014 Macmillan Publishers Limited All rights reserved 0950-9232/14 www.nature.com/onc ORIGINAL ARTICLE CD82/KAI expression prevents IL-8-mediated endothelial gap formation in late-stage melanomas P Khanna1, C-Y Chung2, RI Neves2,3,4,5,6, GP Robertson2,3,4,7,8,9 and C Dong1,8 Melanoma cells facilitate endothelial gap formation, the first step during tumor transendothelial migration, which is mediated by both adhesion and endogenously produced chemokines (in particular, interleukin-8 (IL-8)). Tetraspanins are localized to the cell surface in cancer and participate in various functions including invasion of tissues mediated by secretion of cytokines and matrix metalloproteinases. However, little is known about the role of CD82 tetraspanins in malignant melanomas during cancer cell invasion. In this study, we investigated the functional importance of CD82 expression in melanoma-mediated gap formation by using cDNAs to induce CD82 expression in highly invasive melanoma cell lines. Results showed that CD82 expression inhibited melanoma cell-induced gap formation, melanoma cell extravasation in vitro and subsequent lung metastasis development in vivo. Mechanistic studies showed that inducible expression of CD82 in highly metastatic melanoma cells significantly increased p21 expression upon binding of Duffy antigen receptor group (DARC), inducing tumor cell senescence and interrupting IL-8-mediated vascular endothelial (VE)-cadherin disassembly. Taken together, these studies provide a rationale for using drug therapies
    [Show full text]
  • Project 1: Investigating the Interaction Between the Tetraspanin CD82 And
    Investigating the Interaction between the Tetraspanin CD82 and Gangliosides And Investigating the Binding Partners of CLEC14A and the Function of the CLEC14A Extracellular Domain By Puja Lodhia A dissertation submitted to the University of Birmingham in partial fulfilment of the Degree of MRes Biomedical Research College of Medical and Dental Sciences University Of Birmingham Submitted August 2012 University of Birmingham Research Archive e-theses repository This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder. Investigating the Interaction Between the Tetraspanin CD82 and Gangliosides By Puja Lodhia A dissertation submitted to the University of Birmingham in partial fulfilment of the Degree of MRes Biomedical Research College of Medical and Dental Sciences University Of Birmingham Submitted August 2012 Abstract CD82 is a tetraspanin involved in tumour metastasis suppression. It is downregulated in several cancers including breast, liver and prostate cancer. Although interactions with gangliosides and cholesterol have been demonstrated, no ligand has yet been identified to directly target CD82. CD82 interaction with gangliosides has been explored by immunoprecipitation from cell lysates. However, direct interaction of purified full length CD82 protein with ganglioside has not yet been shown.
    [Show full text]
  • Flow Cytometric Analysis of B-Cell Lymphoproliferative Disorders
    Flow cytometric analysis of B-cell lymphoproliferative disorders David M. Dorfman, M.D., Ph.D. Department of Pathology Brigham and Women’s Hospital and Harvard Medical School Boston, MA Objectives • Review basic principles of flow cytometric immunophenotypic analysis of B cell lymphoproliferative disorders • Discuss recent studies to overcome limitations and shortcomings – New markers – New methods Incidence of B-cell neoplasms, United States Subtype Incidence rate 2011-2012 New cases, 2016 per 100,000 Lymphoid neoplasms 34.4 136,960 Lymphoid neoplasms, B 29.0 93.3% 117,470 B-LL/L 1.4 82.2% 4,930 CLL/SLL 5.1 20,980 FL 3.4 13,960 DLBCL 6.3 27,650 MM 5.9 24,280 Lymphoid neoplasms, T/NK 2.1 8,380 T-LL/L 0.3 1,070 T-PLL <0.1 160 T-LGL 0.2 670 ATL/L <0.1 180 Teras et al. CA Cancer J Clin 2016; 66:443-459 (North American AssociationTeras of et Central al. CA Cancer Cancer Registries) J Clin 2016; 66:443-459 (North American Association of Central Cancer Registries) SS <0.1 Teras et al. CA Cancer J Clin70 2016; 66:443-459 (North American Association of Central Cancer Registries) 94% WHO revised 4th ed., 2017 Flow cytometric analysis of B-cell lymphoproliferative disorders • B-cell antigen expression (CD19, CD20, CD22) • Monoclonal surface immunoglobulin κ or λ light chain expression (or absence of surface immunoglobulin) • Expression of additional B-cell antigens or other antigens, including abnormal expression levels • Presence of cells with abnormal light scatter characteristics ( high forward scatter or side scatter) B-ALL MCL FL, HL MZL, CLL, MM LPL DLBCL DLBCL WHO revised 4th ed.
    [Show full text]
  • B Lymphocyte Binding to E- and P-Selectins Is Mediated Through the De Novo Expression of Carbohydrates on in Vitro and in Vivo Activated Human B Cells
    B lymphocyte binding to E- and P-selectins is mediated through the de novo expression of carbohydrates on in vitro and in vivo activated human B cells. A A Postigo, … , F Sánchez-Madrid, M O de Landázuri J Clin Invest. 1994;94(4):1585-1596. https://doi.org/10.1172/JCI117500. Research Article Cell adhesion to endothelium regulates the trafficking and recruitment of leukocytes towards lymphoid organs and sites of inflammation. This phenomenon is mediated by the expression of a number of adhesion molecules on both the endothelium and circulating cells. Activation of endothelial cells (EC) with different stimuli induces the expression of several adhesion molecules (E- and P-selectins, ICAM-1, VCAM-1), involved in their interaction with circulating cells. In this report, we have studied the binding of nonactivated and activated B cells to purified E- and P-selectins. Activated but not resting B cells were able to interact with both selectins. This binding capacity of activated B cells paralleled the induction of different carbohydrate epitopes (Lewisx, sialyl-Lewisx, CD57 and CDw65) as well as other molecules bearing these or related epitopes in myeloid cells (L-selectin, alpha L beta 2 and alpha X beta 2 integrins, and CD35) involved in the interaction of different cell types with selectins. B cells infiltrating inflamed tissues like in Hashimoto's thyroiditis, also expressed these selectin-binding carbohydrates in parallel with the expression of E-selectin by surrounding follicular dendritic cells. Moreover, the crosslinking of these selectin-binding epitopes resulted in an increased binding of B cells to different integrin ligands.
    [Show full text]
  • CD81 Protein Is Expressed at High Levels in Normal Germinal Center B Cells and in Subtypes of Human Lymphomas☆ Robert F
    Human Pathology (2010) 41, 271–280 www.elsevier.com/locate/humpath Original contribution CD81 protein is expressed at high levels in normal germinal center B cells and in subtypes of human lymphomas☆ Robert F. Luo MD, MPH a, Shuchun Zhao MS a, Robert Tibshirani PhD b, June H. Myklebust PhD c, Mrinmoy Sanyal PhD c, Rosemary Fernandez c, Dita Gratzinger MD, PhD a, Robert J. Marinelli PhD d, Zhi Shun Lu MS a, Anna Wong MD a, Ronald Levy MD c, Shoshana Levy PhD c,1, Yasodha Natkunam MD, PhD a,⁎,1 aDepartment of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA bDepartment of Health Research and Policy and Statistics, Stanford University School of Medicine, Stanford, CA 94305, USA cDivision of Oncology, Department of Medicine, Division of Oncology, Stanford University School of Medicine, Stanford, CA 94305, USA dDepartment of Biochemistry, Stanford University School of Medicine, Stanford, CA 94305, USA Received 8 May 2009; revised 28 July 2009; accepted 30 July 2009 Keywords: Summary CD81 is a tetraspanin cell surface protein that regulates CD19 expression in B lymphocytes CD81; and enables hepatitis C virus infection of human cells. Immunohistologic analysis in normal Lymphoma; hematopoietic tissue showed strong staining for CD81 in normal germinal center B cells, a cell type in Tissue microarray which its increased expression has not been previously recognized. High-dimensional flow cytometry analysis of normal hematopoietic tissue confirmed that among B- and T-cell subsets, germinal center B cells showed the highest level of CD81 expression. In more than 800 neoplastic tissue samples, its expression was also found in most non-Hodgkin lymphomas.
    [Show full text]