DOCSLIB.ORG

Microtubule Dynamics Cytotoxicity by Regulating Actin and Vav1 Controls

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Microtubule Dynamics Cytotoxicity by Regulating Actin and Vav1 Controls Vav1 Controls DAP10-Mediated Natural Cytotoxicity by Regulating Actin and Microtubule Dynamics This information is current as Daniel B. Graham, Marina Cella, Emanuele Giurisato, Keiko of October 1, 2021. Fujikawa, Ana V. Miletic, Tracie Kloeppel, Karry Brim, Toshiyuki Takai, Andrey S. Shaw, Marco Colonna and Wojciech Swat J Immunol 2006; 177:2349-2355; ; doi: 10.4049/jimmunol.177.4.2349 http://www.jimmunol.org/content/177/4/2349 Downloaded from References This article cites 35 articles, 17 of which you can access for free at: http://www.jimmunol.org/content/177/4/2349.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 October 1, 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 © 2006 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Vav1 Controls DAP10-Mediated Natural Cytotoxicity by Regulating Actin and Microtubule Dynamics1 Daniel B. Graham,* Marina Cella,* Emanuele Giurisato,* Keiko Fujikawa,† Ana V. Miletic,* Tracie Kloeppel,* Karry Brim,* Toshiyuki Takai,‡ Andrey S. Shaw,* Marco Colonna,* and Wojciech Swat2* The NK cell-activating receptor NKG2D recognizes several MHC class I-related molecules expressed on virally infected and tumor cells. Human NKG2D transduces activation signals exclusively via an associated DAP10 adaptor containing a YxNM motif, whereas murine NKG2D can signal through either DAP10 or the DAP12 adaptor, which contains an ITAM sequence. DAP10 signaling is thought to be mediated, at least in part, by PI3K and is independent of Syk/Zap-70 kinases; however, the exact mechanism by which DAP10 induces natural cytotoxicity is incompletely understood. Herein, we identify Vav1, a Rho GTPase guanine nucleotide exchange factor, as a critical signaling mediator downstream of DAP10 in NK cells. Specifically, using mice Downloaded from deficient in Vav1 and DAP12, we demonstrate an essential role for Vav1 in DAP10-induced NK cell cytoskeletal polarization involving both actin and microtubule networks, maturation of the cytolytic synapse, and target cell lysis. Mechanistically, we show that Vav1 interacts with DAP10 YxNM motifs through the adaptor protein Grb2 and is required for activation of PI3K-dependent Akt signaling. Based on these findings, we propose a novel model of ITAM-independent signaling by Vav downstream of DAP10 in NK cells. The Journal of Immunology, 2006, 177: 2349–2355. http://www.jimmunol.org/ atural cytotoxicity mediated by NK cells is regulated by Activating NK cell receptors primarily signal through ITAM- multiple activating and inhibitory receptors, which con- containing adaptor molecules such as DAP12, CD3␨, and FcR␥, N fer innate defenses against tumor cells and virus-infected which initiate cellular activation signals by recruiting Syk/Zap-70 cells. The multistep process leading to target cell lysis involves the family kinases (6–9). Additional NK cell activating receptors, formation of a cytolytic synapse and polarized degranulation of the such as NKG2D, trigger cytotoxicity independently of ITAMs by NK cell (1–4). Initial contact between the NK cell and the target associating with DAP10, a unique adaptor containing a YxNM cell is mediated by integrins and facilitates engagement of NK- motif that recruits PI3K (10) and Grb2 (11). Initiation of NKG2D activating receptors by their cognate ligands (2, 4). Postconjuga- signals occurs upon recognition of specific ligands, stress-induced tion events are orchestrated by signals emanating from activating MHC class I-like molecules such as MICA, MICB, and UL-16 by guest on October 1, 2021 receptors and involve F-actin accumulation at the NK-target con- binding protein in humans, as well as Rae-1, H-60, and MULT1 in tact site and microtubule-organizing center (MTOC)3 polarization mice (8, 12). Subsequent to ligand engagement, human NKG2D toward the target cell. In turn, MTOC polarization leads to the signals through DAP10, whereas murine NKG2D signals through establishment of a microtubule network guiding cytolytic granules distinct adaptor molecules. Full-length NKG2D-long (NKG2D-L) to the synapse where they fuse with the plasma membrane and signals through DAP10, whereas a shorter splice variant release perforin and granzymes to lyse target cells (2, 4). Cytoskel- (NKG2D-S) signals through both DAP10 and DAP12 (13–15). In etal remodeling is critical for NK cytotoxicity because pharmaco- this regard, the relative proportion of NKG2D-L and NKG2D-S in logic inhibition of F-actin or microtubule dynamics blocks granule NK cells varies upon in vitro activation with IL-2. Thus, freshly polarization and target cell lysis (1). Moreover, NK cells from isolated (ex vivo) NK cells predominantly express NKG2D-L, patients bearing mutations in WASp that disrupt actin dynamics whereas in vitro activation leads to an increase in NKG2D-S ex- fail to initiate synapse formation and lyse target cells (1, 5). pression (13). Nevertheless, experiments with DAP12-deficient murine NK cells demonstrate that DAP10 is sufficient to mediate NKG2D-dependent cytotoxicity (16). Furthermore, human *Department of Pathology and Immunology, Washington University School of Med- NKG2D promotes NK cytotoxicity despite its inability to interact icine and Siteman Cancer Center, St. Louis, MO 63110; †Department of Pathology and Immunology, Hokkaido University School of Medicine, Sapporo, Japan; and with DAP12 (17). ‡Department of Experimental Immunology, Institute of Development, Aging and Previous studies have implicated the Vav family of Rho guanine Cancer, Tohoku University, Sendai, Japan nucleotide exchange factors in the regulation of several distinct Received for publication March 22, 2006. Accepted for publication May 25, 2006. pathways controlling natural cytotoxicity (18–22). NK cells lack- The costs of publication of this article were defrayed in part by the payment of page ing all three Vav proteins show severely compromised cytotoxicity charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. triggered by both ITAM- and DAP10-associated activating recep- 1 This work is supported by National Institutes of Health Grants R21 AI063024 (to tors (18). However, while deficiency in Vav1 alone primarily im- W.S.) and T32 CA009547 (to D.B.G.). paired the NKG2D-DAP10 cytolytic pathway, lack of Vav2 and 2 Address correspondence and reprint requests to Dr. Wojciech Swat, Department of Vav3 reduced cytotoxicity triggered by receptors that signal Pathology and Immunology, Washington University School of Medicine, 660 South through ITAM-containing adaptors (18). These observations indi- Euclid Avenue, St. Louis, MO 63110. E-mail address: [email protected] cated an unexpected specialization of Vav proteins in regulating 3 Abbreviations used in this paper: MTOC, microtubule-organizing center; NKG2D-L, NKG2D long; NKG2D-S, NKG2D short; SH, Src homology; WT, wild distinct cytotoxic pathways and implicated Vav1 in control of sig- type. nals emanating from DAP10-coupled receptors. However the exact Copyright © 2006 by The American Association of Immunologists, Inc. 0022-1767/06/$02.00 2350 Vav IN CYTOSKELETAL REMODELING BY NKG2D-DAP10 mechanism of Vav1 coupling to DAP10 remains elusive. In addi- Biochemistry tion, a particular issue is how DAP10 controls cytoskeletal remod- Purified splenic NK cells were cultured in IL-2 (1000 U/ml) for 7 days and eling events during the cytolytic response. then starved in serum-free medium for 6 h. NK cells (1.25 ϫ 106/sample) were Herein, we sought to elucidate the mechanism of Vav1 function resuspended in HBSS and incubated on ice with biotinylated anti-NKG2D in DAP10-mediated signaling events that control natural cytotox- (Biolegend) at 1 ␮g/1 ϫ 106 cells. After 15 min, streptavidin (Pierce) was ␮ ϫ 6 icity. Using mice deficient in Vav1 and DAP12, we demonstrate a added at 2 g/1 10 cells, and cells were incubated at 37°C for the indicated time points. Cells were then lysed in radioimmunoprecipitation assay buffer critical function for Vav1 in DAP10-induced PI3K activation, F- and analyzed by Western blotting for phospho-serine 473 Akt (Cell Signaling actin polymerization, and MTOC polarization and provide evi- Technology) or total Akt (Cell Signaling Technology). dence that Vav1 is recruited to DAP10 via Grb2. Conjugate formation Materials and Methods Target cells were stained with CFSE, and NK cells were stained with Mice and NK cell purification hydroethidine. NK cells and targets were pelleted together, gently dis- rupted, and incubated at 37°C for 15 min. The percentage of cells forming Vav1Ϫ/Ϫ and DAP12Ϫ/Ϫ mice have been described elsewhere and were conjugates was determined by FACS. bred to generate Vav1Ϫ/ϪDAP12Ϫ/Ϫ double knockout mice (F6 on C57BL/6 background) (23, 24). Splenic NK cells were purified by pos- NK
Load more

Recommended publications
  • Deregulated Gene Expression Pathways in Myelodysplastic Syndrome Hematopoietic Stem Cells
    Leukemia (2010) 24, 756–764 & 2010 Macmillan Publishers Limited All rights reserved 0887-6924/10 $32.00 www.nature.com/leu ORIGINAL ARTICLE Deregulated gene expression pathways in myelodysplastic syndrome hematopoietic stem cells A Pellagatti1, M Cazzola2, A Giagounidis3, J Perry1, L Malcovati2, MG Della Porta2,MJa¨dersten4, S Killick5, A Verma6, CJ Norbury7, E Hellstro¨m-Lindberg4, JS Wainscoat1 and J Boultwood1 1LRF Molecular Haematology Unit, NDCLS, John Radcliffe Hospital, Oxford, UK; 2Department of Hematology Oncology, University of Pavia Medical School, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy; 3Medizinische Klinik II, St Johannes Hospital, Duisburg, Germany; 4Division of Hematology, Department of Medicine, Karolinska Institutet, Stockholm, Sweden; 5Department of Haematology, Royal Bournemouth Hospital, Bournemouth, UK; 6Albert Einstein College of Medicine, Bronx, NY, USA and 7Sir William Dunn School of Pathology, University of Oxford, Oxford, UK To gain insight into the molecular pathogenesis of the the World Health Organization.6,7 Patients with refractory myelodysplastic syndromes (MDS), we performed global gene anemia (RA) with or without ringed sideroblasts, according to expression profiling and pathway analysis on the hemato- poietic stem cells (HSC) of 183 MDS patients as compared with the the French–American–British classification, were subdivided HSC of 17 healthy controls. The most significantly deregulated based on the presence or absence of multilineage dysplasia. In pathways in MDS include interferon signaling, thrombopoietin addition, patients with RA with excess blasts (RAEB) were signaling and the Wnt pathways. Among the most signifi- subdivided into two categories, RAEB1 and RAEB2, based on the cantly deregulated gene pathways in early MDS are immuno- percentage of bone marrow blasts.
    [Show full text]
  • The Multifaceted Role of CMA in Glioma: Enemy Or Ally?
    International Journal of Molecular Sciences Review The Multifaceted Role of CMA in Glioma: Enemy or Ally? Alessia Lo Dico 1,† , Cristina Martelli 1,† , Cecilia Diceglie 1 and Luisa Ottobrini 1,2,* 1 Department of Pathophysiology and Transplantation, University of Milan, Via F.Cervi 93, Segrate, 20090 Milan, Italy; [email protected] (A.L.D.); [email protected] (C.M.); [email protected] (C.D.) 2 Institute of Molecular Bioimaging and Physiology, National Research Council (IBFM-CNR), Via F.Cervi 93, Segrate, 20090 Milan, Italy * Correspondence: [email protected]; Tel.: +39-02(50)-330-404; Fax: +39-02(50)-330-425 † Authors contributed equally to this work. Abstract: Chaperone-mediated autophagy (CMA) is a catabolic pathway fundamental for cell home- ostasis, by which specific damaged or non-essential proteins are degraded. CMA activity has three main levels of regulation. The first regulatory level is based on the targetability of specific proteins possessing a KFERQ-like domain, which can be recognized by specific chaperones and delivered to the lysosomes. Target protein unfolding and translocation into the lysosomal lumen constitutes the second level of CMA regulation and is based on the modulation of Lamp2A multimerization. Finally, the activity of some accessory proteins represents the third regulatory level of CMA activity. CMA’s role in oncology has not been fully clarified covering both pro-survival and pro-death roles in different contexts. Taking all this into account, it is possible to comprehend the actual complexity of both CMA regulation and the cellular consequences of its activity allowing it to be elected as a modulatory and not only catabolic machinery.
    [Show full text]
  • Mutation of Vav1 Adaptor Region Reveals a New Oncogenic Activation
    www.impactjournals.com/oncotarget/ Oncotarget, Vol. 6, No.4 Mutation of Vav1 adaptor region reveals a new oncogenic activation Lyra Razanadrakoto1,2,*, Françoise Cormier3,4,5,*, Vanessa Laurienté1,2, Elisabetta Dondi1,2, Laura Gardano1,2, Shulamit Katzav6, Lionel Guittat1,2 and Nadine Varin- Blank1,2 1 INSERM, UMR 978, Bobigny, France 2 PRES SPC, Labex Inflamex, Université Paris 13, UFR SMBH, Bobigny, France 3 INSERM, UMR 1016, Institut Cochin, Paris, France 4 CNRS, UMR 8104, Paris, France 5 PRES SPC, Université Paris Descartes, Paris, France 6 The Hebrew University/ Hadassah Medical School, Jerusalem, Israel * These authors share co-first authorship Correspondence to: Nadine Varin-Blank, email: [email protected] Correspondence to: Lionel Guittat, email: [email protected] Keywords: Vav1, β-catenin, Rac GTPase, Src-homology domains, adhesion complex, tumorigenesis Received: May 14, 2014 Accepted: October 23, 2014 Published: October 24 2014 This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ABSTRACT Vav family members function as remarkable scaffold proteins that exhibit both GDP/GTP exchange activity for Rho/Rac GTPases and numerous protein-protein interactions via three adaptor Src-homology domains. The exchange activity is under the unique regulation by phosphorylation of tyrosine residues hidden by intra-molecular interactions. Deletion of the autoinhibitory N-terminal region results in an oncogenic protein, onco-Vav, leading to a potent activation of Rac GTPases whereas the proto-oncogene barely leads to transformation. Substitution of conserved residues of the SH2-SH3 adaptor region in onco-Vav reverses oncogenicity.
    [Show full text]
  • Vav1 (D45G3) Rabbit Mab A
    Revision 1 C 0 2 - t Vav1 (D45G3) Rabbit mAb a e r o t S Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) 7 5 Web: [email protected] 6 www.cellsignal.com 4 # 3 Trask Lane Danvers Massachusetts 01923 USA For Research Use Only. Not For Use In Diagnostic Procedures. Applications: Reactivity: Sensitivity: MW (kDa): Source/Isotype: UniProt ID: Entrez-Gene Id: WB, IP H Endogenous 95 Rabbit IgG P15498 7409 Product Usage Information Application Dilution Western Blotting 1:1000 Immunoprecipitation 1:50 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody. Specificity / Sensitivity Vav1 (D45G3) Rabbit mAb recognizes endogenous levels of total Vav1 protein. Species Reactivity: Human Species predicted to react based on 100% sequence homology: Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues in the carboxy terminus of human Vav1 protein. Background Vav proteins belong to the Dbl family of guanine nucleotide exchange factors (GEFs) for Rho/Rac small GTPases. The three identified mammalian Vav proteins (Vav1, Vav2 and Vav3) differ in their expression. Vav1 is expressed only in hematopoietic cells and is involved in the formation of the immune synapse. Vav2 and Vav3 are more ubiquitously expressed. Vav proteins contain the Dbl homology domain, which confers GEF activity, as well as protein interaction domains that allow them to function in pathways regulating actin cytoskeleton organization (reviewed in 1).
    [Show full text]
  • MERTK-Mediated Signaling in the Retinal Pigment Epithelium: Insights Into the Mechanism of RPE Phagocytosis
    MERTK-mediated Signaling in the Retinal Pigment Epithelium: Insights into the Mechanism of RPE Phagocytosis by Shameka J. Shelby A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Biological Chemistry) in The University of Michigan 2012 Doctoral Committee: Professor Debra A. Thompson, Chair Professor Christin Carter-Su Professor Bret A. Hughes Professor Benjamin L. Margolis Assistant Professor Hisashi Umemori © Shameka J. Shelby 2012 To my two favorite men, Juan and Zani for your patience, love, and support. ii Acknowledgements I would like to acknowledge all of the people who helped me to accomplish my goals. My mentor, Dr. Debra Thompson, has been instrumental in my success as a scientist. Her motherly guidance has helped to mold me into the person and scientist I am today. I’m overly grateful for her tireless efforts and patience during the course of my graduate career. I wouldn’t have even considered graduate school without the guidance of my undergraduate advisors, Drs. Marion Carroll and Guangdi Wang, to whom I am also grateful. Also, my favorite high school teacher Mrs. Jenkins incited my passion for biology and encouraged me to follow my aspirations. I am also grateful for the invaluable input and criticisms of my committee members, Drs. Bret Hughes, Christin Carter-Su, Hisashi Umemori, and Benjamin Margolis. I would especially like to thank Dr. Hughes, Dr. Carter-Su, and Dr. Margolis for reagents that were instrumental in the completion of my experiments. Ben literally saved my life with the plethora of antibodies he provided. Reagents and clones provided by Drs.
    [Show full text]
  • VAV Proteins As Double Agents in Cancer: Oncogenes with Tumor Suppressor Roles
    biology Review VAV Proteins as Double Agents in Cancer: Oncogenes with Tumor Suppressor Roles Myriam Cuadrado 1,2 and Javier Robles-Valero 1,2,* 1 Centro de Investigación del Cáncer, CSIC-University of Salamanca, 37007 Salamanca, Spain; [email protected] 2 Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), CSIC-University of Salamanca, 37007 Salamanca, Spain * Correspondence: [email protected] Simple Summary: The role of the VAV family (comprised of VAV1, VAV2, and VAV3) in proactive pathways involved in cell transformation has been historically assumed. Indeed, the discovery of potential gain-of-function VAV1 mutations in specific tumor subtypes reinforced this functional archetype. Contrary to this paradigm, we demonstrated that VAV1 could unexpectedly act as a tumor suppressor in some in vivo contexts. In this review, we discuss recent findings in the field, where the emerging landscape is one in which GTPases and their regulators, such as VAV proteins, can exhibit tumor suppressor functions. Abstract: Guanosine nucleotide exchange factors (GEFs) are responsible for catalyzing the transition of small GTPases from the inactive (GDP-bound) to the active (GTP-bound) states. RHO GEFs, including VAV proteins, play essential signaling roles in a wide variety of fundamental cellular processes and in human diseases. Although the most widespread archetype in the field is that RHO Citation: Cuadrado, M.; Robles-Valero, GEFs exert proactive functions in cancer, recent studies in mice and humans are providing new J. VAV Proteins as Double Agents in insights into the in vivo function of these proteins in cancer. These results suggest a more complex Cancer: Oncogenes with Tumor scenario where the role of GEFs is not so clearly defined.
    [Show full text]
  • The Role of the Src Homology-2 Domain Containing Protein B (SHB) in B Cells
    M WELSH and others Role of SHB in b cells 56:1 R21–R31 Review The role of the Src Homology-2 domain containing protein B (SHB) in b cells Correspondence † Michael Welsh, Maria Jamalpour, Guangxiang Zang and Bjo¨rn Åkerblom should be addressed to M Welsh Department of Medical Cell Biology, Uppsala University, PO Box 571, Husargatan 3, SE-75123 Uppsala, Sweden Email †G Zang is now at Department of Medical Biosciences, Umea˚ University, Umea˚ , Sweden [email protected] Abstract This review will describe the SH2-domain signaling protein Src Homology-2 domain Key Words containing protein B (SHB) and its role in various physiological processes relating in " signal transduction particular to glucose homeostasis and b cell function. SHB operates downstream of several " insulin secretion tyrosine kinase receptors and assembles signaling complexes in response to receptor " vascular activation by interacting with other signaling proteins via its other domains (proline-rich, " islet cells phosphotyrosine-binding and tyrosine-phosphorylation sites). The subsequent responses " immune system are context-dependent. Absence of Shb in mice has been found to exert effects on hematopoiesis, angiogenesis and glucose metabolism. Specifically, first-phase insulin secretion in response to glucose was impaired and this effect was related to altered characteristics of focal adhesion kinase activation modulating signaling through Akt, ERK, b catenin and cAMP. It is believed that SHB plays a role in integrating adaptive responses to various stimuli by simultaneously modulating cellular responses in different cell-types, thus Journal of Molecular Journal of Molecular Endocrinology playing a role in maintaining physiological homeostasis. Endocrinology (2016) 56, R21–R31 Introduction The limited replicative capacity of human pancreatic (bTC1 cells) after serum-stimulation, the Src Homology-2 b cells contributes significantly to the development of domain containing adapter protein B (SHB) was identified diabetes mellitus and thus comprises a major hurdle for (Welsh et al.
    [Show full text]
  • The Human Gene Connectome As a Map of Short Cuts for Morbid Allele Discovery
    The human gene connectome as a map of short cuts for morbid allele discovery Yuval Itana,1, Shen-Ying Zhanga,b, Guillaume Vogta,b, Avinash Abhyankara, Melina Hermana, Patrick Nitschkec, Dror Friedd, Lluis Quintana-Murcie, Laurent Abela,b, and Jean-Laurent Casanovaa,b,f aSt. Giles Laboratory of Human Genetics of Infectious Diseases, Rockefeller Branch, The Rockefeller University, New York, NY 10065; bLaboratory of Human Genetics of Infectious Diseases, Necker Branch, Paris Descartes University, Institut National de la Santé et de la Recherche Médicale U980, Necker Medical School, 75015 Paris, France; cPlateforme Bioinformatique, Université Paris Descartes, 75116 Paris, France; dDepartment of Computer Science, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel; eUnit of Human Evolutionary Genetics, Centre National de la Recherche Scientifique, Unité de Recherche Associée 3012, Institut Pasteur, F-75015 Paris, France; and fPediatric Immunology-Hematology Unit, Necker Hospital for Sick Children, 75015 Paris, France Edited* by Bruce Beutler, University of Texas Southwestern Medical Center, Dallas, TX, and approved February 15, 2013 (received for review October 19, 2012) High-throughput genomic data reveal thousands of gene variants to detect a single mutated gene, with the other polymorphic genes per patient, and it is often difficult to determine which of these being of less interest. This goes some way to explaining why, variants underlies disease in a given individual. However, at the despite the abundance of NGS data, the discovery of disease- population level, there may be some degree of phenotypic homo- causing alleles from such data remains somewhat limited. geneity, with alterations of specific physiological pathways under- We developed the human gene connectome (HGC) to over- come this problem.
    [Show full text]
  • SYK Expression in Monomorphic Epitheliotropic Intestinal T-Cell
    Modern Pathology (2018) 31, 505–516 © 2018 USCAP, Inc All rights reserved 0893-3952/18 $32.00 505 SYK expression in monomorphic epitheliotropic intestinal T-cell lymphoma Grit Mutzbauer1, Katja Maurus1, Clara Buszello1, Jordan Pischimarov1, Sabine Roth1, Andreas Rosenwald1,2, Andreas Chott3 and Eva Geissinger1,2 1Institute of Pathology, University of Wuerzburg, Wuerzburg, Germany; 2Comprehensive Cancer Center Mainfranken, University and University Hospital, Wuerzburg, Germany and 3Institute of Pathology and Microbiology, Wilhelminenspital, Vienna, Austria Monomorphic epitheliotropic intestinal T-cell lymphoma (MEITL), formerly known as type II enteropathy associated T-cell lymphoma (type II EATL), is a rare, aggressive primary intestinal T-cell lymphoma with a poor prognosis and an incompletely understood pathogenesis. We collected 40 cases of MEITL and 27 cases of EATL, formerly known as type I EATL, and comparatively investigated the T-cell receptor (TCR) itself and associated signaling molecules using immunohistochemistry, amplicon deep sequencing and bisulfite pyrosequencing. The TCR showed both an αβ-T-cell origin (30%) and a γδ-T-cell derivation (55%) resulting in a predominant positive TCR phenotype in MEITL compared with the mainly silent TCR phenotype in EATL (65%). The immunohisto- chemical expression of the spleen tyrosine kinase (SYK) turned out to be a distinctive feature of MEITL (95%) compared with EATL (0%). Aberrant SYK overexpression in MEITL is likely caused by hypomethylation of the SYK promoter, while no common mutations in the SYK gene or in its promoter could be detected. Using amplicon deep sequencing, mutations in DNMT3A, IDH2, and TET2 were infrequent events in MEITL and EATL. Immunohistochemical expression of linker for activation of T-cells (LAT) subdivided MEITL into a LAT expressing subset (33%) and a LAT silent subset (67%) with a potentially earlier disease onset in LAT-positive MEITL.
    [Show full text]
  • Oral Administration of Lactobacillus Plantarum 299V
    Genes Nutr (2015) 10:10 DOI 10.1007/s12263-015-0461-7 RESEARCH PAPER Oral administration of Lactobacillus plantarum 299v modulates gene expression in the ileum of pigs: prediction of crosstalk between intestinal immune cells and sub-mucosal adipocytes 1 1,4 1,5 1 Marcel Hulst • Gabriele Gross • Yaping Liu • Arjan Hoekman • 2 1,3 1,3 Theo Niewold • Jan van der Meulen • Mari Smits Received: 19 November 2014 / Accepted: 28 March 2015 / Published online: 11 April 2015 Ó The Author(s) 2015. This article is published with open access at Springerlink.com Abstract To study host–probiotic interactions in parts of ileum. A higher expression level of several B cell-specific the intestine only accessible in humans by surgery (je- transcription factors/regulators was observed, suggesting junum, ileum and colon), pigs were used as model for that an influx of B cells from the periphery to the ileum humans. Groups of eight 6-week-old pigs were repeatedly and/or the proliferation of progenitor B cells to IgA-com- orally administered with 5 9 1012 CFU Lactobacillus mitted plasma cells in the Peyer’s patches of the ileum was plantarum 299v (L. plantarum 299v) or PBS, starting with stimulated. Genes coding for enzymes that metabolize a single dose followed by three consecutive daily dosings leukotriene B4, 1,25-dihydroxyvitamin D3 and steroids 10 days later. Gene expression was assessed with pooled were regulated in the ileum. Bioinformatics analysis pre- RNA samples isolated from jejunum, ileum and colon dicted that these metabolites may play a role in the scrapings of the eight pigs per group using Affymetrix crosstalk between intestinal immune cells and sub-mucosal porcine microarrays.
    [Show full text]
  • JAK Tyrosine Kinases Promote Hierarchical Activation of Rho and Rap Modules of Integrin Activation
    Published Online: 23 December, 2013 | Supp Info: http://doi.org/10.1083/jcb.201303067 JCB: Article Downloaded from jcb.rupress.org on May 12, 2019 JAK tyrosine kinases promote hierarchical activation of Rho and Rap modules of integrin activation Alessio Montresor,1,2 Matteo Bolomini-Vittori,1 Lara Toffali,1 Barbara Rossi,1 Gabriela Constantin,1 and Carlo Laudanna1,2 1Department of Pathology and Diagnostics, Division of General Pathology, School of Medicine, and 2The Center for Biomedical Computing, University of Verona, Verona 37134, Italy ymphocyte recruitment is regulated by signaling is dependent on JAK activity, with VAV1 mediating Rho modules based on the activity of Rho and Rap small activation by JAKs in a Gi-independent manner. Fur- L guanosine triphosphatases that control integrin acti- thermore, activation of Rap1A by chemokines is also vation by chemokines. We show that Janus kinase (JAK) dependent on JAK2 and JAK3 activity. Importantly, ac- protein tyrosine kinases control chemokine-induced LFA-1– tivation of Rap1A by JAKs is mediated by RhoA and and VLA-4–mediated adhesion as well as human T lym- PLD1, thus establishing Rap1A as a downstream effector phocyte homing to secondary lymphoid organs. JAK2 of the Rho module. Thus, JAK tyrosine kinases control and JAK3 isoforms, but not JAK1, mediate CXCL12- integrin activation and dependent lymphocyte trafficking induced LFA-1 triggering to a high affinity state. Signal by bridging chemokine receptors to the concurrent and transduction analysis showed that chemokine-induced hierarchical activation of the Rho and Rap modules of activation of the Rho module of LFA-1 affinity triggering integrin activation.
    [Show full text]
  • Two Closely Spaced Tyrosines Regulate NFAT Signaling in B Cells Via Syk Association with Vavᰔ Chih-Hong Chen, Victoria A
    MOLECULAR AND CELLULAR BIOLOGY, July 2011, p. 2984–2996 Vol. 31, No. 14 0270-7306/11/$12.00 doi:10.1128/MCB.05043-11 Copyright © 2011, American Society for Microbiology. All Rights Reserved. Two Closely Spaced Tyrosines Regulate NFAT Signaling in B Cells via Syk Association with Vavᰔ Chih-Hong Chen, Victoria A. Martin, Nina M. Gorenstein, Robert L. Geahlen, and Carol Beth Post* Department of Medicinal Chemistry and Molecular Pharmacology, Markey Center for Structural Biology and Purdue Center for Cancer Research, Purdue University, West Lafayette, Indiana 47907 Received 12 January 2011/Returned for modification 16 February 2011/Accepted 12 May 2011 Activated Syk, an essential tyrosine kinase in B cell signaling, interacts with Vav guanine nucleotide exchange factors and regulates Vav activity through tyrosine phosphorylation. The Vav SH2 domain binds Syk linker B by an unusual recognition of two closely spaced Syk tyrosines: Y342 and Y346. The binding affinity is highest when both Y342 and Y346 are phosphorylated. An investigation in B cells of the dependence of Vav phosphorylation and NFAT activation on phosphorylation of Y342 and Y346 finds that cellular response levels match the relative binding affinities of the Vav1 SH2 domain for singly and doubly phosphorylated linker B peptides. This key result suggests that the uncommon recognition determinant of these two closely spaced tyrosines is a limiting factor in signaling. Interestingly, differences in affinities for binding singly and doubly phosphorylated peptides are reflected in the on rate, not the off rate. Such a control mechanism would be highly effective for regulating binding among competing Syk binding partners.
    [Show full text]