DOCSLIB.ORG

Integrins 2 Β and 1Β Through Differential Regulation of Promotion

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Integrins 2 Β and 1Β Through Differential Regulation of Promotion Promotion of Neutrophil Chemotaxis Through Differential Regulation of β1 and β 2 Integrins This information is current as Mary Beth Harler, Eric Wakshull, Edward J. Filardo, Jorge of October 1, 2021. E. Albina and Jonathan S. Reichner J Immunol 1999; 162:6792-6799; ; http://www.jimmunol.org/content/162/11/6792 Downloaded from References This article cites 44 articles, 29 of which you can access for free at: http://www.jimmunol.org/content/162/11/6792.full#ref-list-1 Why The JI? Submit online. http://www.jimmunol.org/ • 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 by guest on October 1, 2021 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 © 1999 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Promotion of Neutrophil Chemotaxis Through Differential b b 1 Regulation of 1 and 2 Integrins Mary Beth Harler,* Eric Wakshull,† Edward J. Filardo,* Jorge E. Albina,* and Jonathan S. Reichner2* b b Migration of neutrophils requires sequential adhesive and deadhesive interactions between 1 and 2 integrins and components b b of the extracellular matrix. Prompted by reports that describe interaction of soluble -glucan with the 2 integrin Mac-1, a role for b-glucan in regulation of integrin-mediated migration was investigated. Neutrophil migration in response to fMLP was assessed using an agarose overlay method with slides precoated with fibronectin (Fn) 6 b-glucan. On Fn, random migration in excess of directed migration was observed. In contrast, migration on Fn 1 b-glucan was directional, with marked diminution of random migration. This conversion of random to directed migration was seen neither when Fn was supplemented with alternative polysaccharides nor when b-glucan was applied to other components of the extracellular matrix. This effect of b-glucan was shown Downloaded from to be cation dependent and to be effected by Arg-Gly-Asp-containing peptides consistent with an integrin-mediated event. mAb inhibition studies demonstrate that b-glucan effects this shift toward directed migration through suppression of migration me- diated by Mac-1 and very late Ag 5 and enhancement of very late Ag 3-mediated migration. Adhesion assays suggest that the prochemotactic influence of b-glucan is due, in part but not entirely, to modulation of PMN adhesion to Fn. In summary, these b b b data support a novel role for -glucan in regulation of 1- and 2-mediated neutrophil migration on Fn. The Journal of Immunology, 1999, 162: 6792–6799. http://www.jimmunol.org/ b ocalization of circulatory polymorphonuclear leukocytes of neutrophil locomotion include members of the 2 (LFA-1 and 3 b (PMN) to extravascular sites of inflammation is a func- Mac-1) (6–8) and 1 subfamilies (VLA-4, -5, and -6) (9, 10). L tion of repeated adhesive and deadhesive events. The se- The mechanisms that regulate the recruitment of extravasated quential processes of PMN rolling, adhesion, and transendothelial PMN to foci of inflammation are not well established. Regulation migration are facilitated by cell surface adhesion receptors referred of integrin function by adhesive substrates offers a mechanism for to as selectins and integrins (1). Following extravasation, PMN local control of migrant cells. Within the assembled framework of migrate toward a source of inflammation in response to locally the ECM, binding sites for integrins have been identified on col- elaborated chemotaxins and cytokines (2). Stimulated by a che- lagen (11), laminin (12), and fibronectin (Fn) (13) among others. by guest on October 1, 2021 motactic gradient, PMN traverse the extracellular matrix (ECM) Because cell migration is the product of sequential adhesive and by way of transient interactions between integrin receptors and deadhesive events, those ligands that permit efficient forward at- components of the ECM that serve as adhesive ligands. Activation tachment and rearward detachment should theoretically serve as of specific integrins through ligand binding has been shown to optimal substrates. augment cell adhesion and precipitate reorganization of the actin Mac-1 (CD11b/CD18, CR3, Mo-1) is a multifunctional receptor cytoskeleton (3). Polarization of the migrant cell ensues, with de- most prominently expressed on myeloid and NK cells (14, 15). velopment of lamellopodia and filopodia (4). Traction at the lead- Structurally classified as an adhesion molecule, with particular rel- ing edge of the cell develops through integrin engagement, fol- evance to cell-mediated interaction with ECM, recent investiga- lowed by translocation of the cell over the adherent segment of tions identify Mac-1 participation in a variety of cellular functions. plasma membrane. Having been shifted to the rear of the cell, the Activities facilitated by Mac-1 include homotypic aggregation (7), integrin then releases its substrate, permitting the cell mass to ad- adhesion (16), migration (8), and binding and phagocytosis of op- vance (5). Integrin receptors reported to contribute to the process sonized microbial pathogens (17, 18). The versatility of Mac-1 is related to its ability to bind an array of soluble and insoluble li- *Department of Surgery, Division of Surgical Research, Rhode Island Hospital and gands, including zymosan (19), fibrinogen (20), ICAM-1 (21), Brown University School of Medicine, Providence, RI, 02903; and †Alpha-Beta Technology, Inc., Worcester, MA iC3b (22), factor X (23), and heparin sulfate (24). Mac-1 is unique b Received for publication December 1, 1998. Accepted for publication March among the 2 integrins in that it also contains a lectin site capable 17, 1999. of interacting with N-acetyl-D-glucosamine (19), glucose, and pos- The costs of publication of this article were defrayed in part by the payment of page sibly mannose (25) residues. Moreover, recent reports describe charges. This article must therefore be hereby marked advertisement in accordance regulation of Mac-1 function through binding of soluble b-glucan with 18 U.S.C. Section 1734 solely to indicate this fact. to the lectin site (26). Specifically, b-glucan has been shown to 1 This work was supported by National Institutes of Health Grants GM-51493 (J.S.R.) and GM-42859 (J.E.A.), by the Surgical Infection Society/Pfizer Fellowship induce cytotoxicity in NK cells for iC3b-opsonized target cells that (M.B.H.), and by allocations to the Department of Surgery by Rhode Island Hospital. were otherwise resistant to killing. 2 Address correspondence and reprint requests to Dr. Jonathan Reichner, Department The b-glucans are a class of long chain polymers of glucose in of Surgery, Rhode Island Hospital, 593 Eddy Street, Providence, RI 02903. E-mail b , linkages that comprise the cell wall of yeast and fungi. Sol- address: [email protected] 1 3 uble b-glucan can be detected in the sera of patients suffering deep 3 Abbreviations used in this paper: PMN, polymorphonuclear leukocyte; ECM, ex- tracellular matrix; Fn, fibronectin; VLA, very late Ag protein; MEM, minimal essen- mycotic infections and febrile episodes due to fungemia (27). tial medium; dpm, disintegrations per minute. Originally identified as the component of zymosan responsible for Copyright © 1999 by The American Association of Immunologists 0022-1767/99/$02.00 The Journal of Immunology 6793 macrophage activation (28), b-glucans have been shown to stim- 72 h in the dark, dialyzed against water and reductively radiolabeled with 3 3 b ulate hemopoietic immune effector cells in both in vivo and in vitro 100 mCi of NaB H4 (New England Nuclear, Boston, MA). The [ H] - glucan was purified by dialysis and ultrafiltration and achieved a sp. act. of models (29, 30). However, the mechanism by which leukocytes 3 6 m 3 5 b 6.8 10 dpm/ g hexose. Approximately 2 10 dpm were added to 100 recognize and respond to soluble -glucan has not been deter- mg cold b-glucan for use as an indicator of binding efficiency to matrix mined. Based upon evidence that interaction of soluble b-glucan components coated onto chambered slides as described above. Following with Mac-1 results in altered effector cell behavior, the possibility incubation for 30 min at 37°C, slides were rinsed to remove unbound 3 that the regulatory effect of b-glucan extends to additional inte- b-glucan. Remaining b-glucan was eluted using 1% SDS and [ H]b-glucan quantitated by liquid scintillation counting. grin-mediated functions was examined. The current investigation tests the hypothesis that, in the context of extracellular matrix, Migration assays b -glucan alters the migratory behavior of neutrophils. Molten agarose (Seakem GTG, FMC Bioproducts, Rockland, ME) was prepared by a modification of the method described (33). Briefly, 1% aga- Materials and Methods rose was boiled in sterile, endotoxin-free sterile saline, followed by 1:1 Isolation of neutrophils dilution with MEM to achieve a final agarose concentration of 0.5% that was then distributed into the precoated chambered slides. Using a plastic Human neutrophils were isolated from the peripheral venous blood of template and beveled punch, three 2-mm wells were created, each sepa- healthy volunteers in heparinized Vacutainer tubes (Becton Dickinson, rated by a distance of 2 mm. The agarose plugs were removed with gentle Lincoln Park, NJ). Granulocytes were prepared by gradient centrifugation aspiration. The central well received 20 ml of prepared cells in MEM at a on Ficoll-Hypaque (Sigma, St.
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]
  • Antibody-Dependent Cellular Cytotoxicity Riiia and Mediate Γ
    Effector Memory αβ T Lymphocytes Can Express Fc γRIIIa and Mediate Antibody-Dependent Cellular Cytotoxicity This information is current as Béatrice Clémenceau, Régine Vivien, Mathilde Berthomé, of September 27, 2021. Nelly Robillard, Richard Garand, Géraldine Gallot, Solène Vollant and Henri Vié J Immunol 2008; 180:5327-5334; ; doi: 10.4049/jimmunol.180.8.5327 http://www.jimmunol.org/content/180/8/5327 Downloaded from References This article cites 43 articles, 21 of which you can access for free at: http://www.jimmunol.org/content/180/8/5327.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 27, 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 © 2008 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Effector Memory ␣␤ T Lymphocytes Can Express Fc␥RIIIa and Mediate Antibody-Dependent Cellular Cytotoxicity1 Be´atrice Cle´menceau,*† Re´gine Vivien,*† Mathilde Berthome´,*† Nelly Robillard,‡ Richard Garand,‡ Ge´raldine Gallot,*† Sole`ne Vollant,*† and Henri Vie´2*† Human memory T cells are comprised of distinct populations with different homing potential and effector functions: central memory T cells that mount recall responses to Ags in secondary lymphoid organs, and effector memory T cells that confer immediate protection in peripheral tissues.
    [Show full text]
  • Epha Receptors and Ephrin-A Ligands Are Upregulated by Monocytic
    Mukai et al. BMC Cell Biology (2017) 18:28 DOI 10.1186/s12860-017-0144-x RESEARCHARTICLE Open Access EphA receptors and ephrin-A ligands are upregulated by monocytic differentiation/ maturation and promote cell adhesion and protrusion formation in HL60 monocytes Midori Mukai, Norihiko Suruga, Noritaka Saeki and Kazushige Ogawa* Abstract Background: Eph signaling is known to induce contrasting cell behaviors such as promoting and inhibiting cell adhesion/ spreading by altering F-actin organization and influencing integrin activities. We have previously demonstrated that EphA2 stimulation by ephrin-A1 promotes cell adhesion through interaction with integrins and integrin ligands in two monocyte/ macrophage cell lines. Although mature mononuclear leukocytes express several members of the EphA/ephrin-A subclass, their expression has not been examined in monocytes undergoing during differentiation and maturation. Results: Using RT-PCR, we have shown that EphA2, ephrin-A1, and ephrin-A2 expression was upregulated in murine bone marrow mononuclear cells during monocyte maturation. Moreover, EphA2 and EphA4 expression was induced, and ephrin-A4 expression was upregulated, in a human promyelocytic leukemia cell line, HL60, along with monocyte differentiation toward the classical CD14++CD16− monocyte subset. Using RT-PCR and flow cytometry, we have also shown that expression levels of αL, αM, αX, and β2 integrin subunits were upregulated in HL60 cells along with monocyte differentiation while those of α4, α5, α6, and β1 subunits were unchanged. Using a cell attachment stripe assay, we have shown that stimulation by EphA as well as ephrin-A, likely promoted adhesion to an integrin ligand- coated surface in HL60 monocytes. Moreover, EphA and ephrin-A stimulation likely promoted the formation of protrusions in HL60 monocytes.
    [Show full text]
  • Slan Monocytes and Macrophages Mediate CD20-Dependent B-Cell
    Published OnlineFirst May 10, 2018; DOI: 10.1158/0008-5472.CAN-17-2344 Cancer Tumor Biology and Immunology Research slanþ Monocytes and Macrophages Mediate CD20-Dependent B-cell Lymphoma Elimination via ADCC and ADCP William Vermi1,2, Alessandra Micheletti3, Giulia Finotti3, Cristina Tecchio4, Federica Calzetti3, Sara Costa3, Mattia Bugatti1, Stefano Calza5, Claudio Agostinelli6, Stefano Pileri7, Piera Balzarini1, Alessandra Tucci8, Giuseppe Rossi8, Lara Furlani4, Giuseppe Todeschini4, Alberto Zamo9, Fabio Facchetti1, Luisa Lorenzi1, Silvia Lonardi1, and Marco A. Cassatella3 Abstract þ Terminal tissue differentiation and function of slan monocytes in cancer þ + + is largely unexplored. Our recent studies demonstrated that slan mono- slan monocyte slan macrophage cytes differentiate into a distinct subset of dendritic cells (DC) in human CD16A þ tonsils and that slan cells colonize metastatic carcinoma-draining lymph CD32 nodes. Herein, we report by retrospective analysis of multi-institutional þ CD16A CD64 cohorts that slan cells infiltrate various types of non-Hodgkin lymphomas = RTX (NHL), particularly the diffuse large B-cell lymphoma (DLBCL) group, CD20 þ CD20 including the most aggressive, nodal and extranodal, forms. Nodal slan cells displayed features of either immature DC or macrophages, in the latter case ingesting tumor cells and apoptotic bodies. We also found in patients þ þ with DLBCL that peripheral blood slan monocytes, but not CD14 monocytes, increased in number and displayed highly efficient rituxi- Lymphoma cell Lymphoma cell mab-mediated antibody-dependent cellular cytotoxicity, almost equivalent + RTX þ to that exerted by NK cells. Notably, slan monocytes cultured in condi- tioned medium from nodal DLBCL (DCM) acquired a macrophage-like + RTX phenotype, retained CD16 expression, and became very efficient in ritux- imab-mediated antibody-dependent cellular phagocytosis (ADCP).
    [Show full text]
  • Supplementary Table 1: Adhesion Genes Data Set
    Supplementary Table 1: Adhesion genes data set PROBE Entrez Gene ID Celera Gene ID Gene_Symbol Gene_Name 160832 1 hCG201364.3 A1BG alpha-1-B glycoprotein 223658 1 hCG201364.3 A1BG alpha-1-B glycoprotein 212988 102 hCG40040.3 ADAM10 ADAM metallopeptidase domain 10 133411 4185 hCG28232.2 ADAM11 ADAM metallopeptidase domain 11 110695 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 195222 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 165344 8751 hCG20021.3 ADAM15 ADAM metallopeptidase domain 15 (metargidin) 189065 6868 null ADAM17 ADAM metallopeptidase domain 17 (tumor necrosis factor, alpha, converting enzyme) 108119 8728 hCG15398.4 ADAM19 ADAM metallopeptidase domain 19 (meltrin beta) 117763 8748 hCG20675.3 ADAM20 ADAM metallopeptidase domain 20 126448 8747 hCG1785634.2 ADAM21 ADAM metallopeptidase domain 21 208981 8747 hCG1785634.2|hCG2042897 ADAM21 ADAM metallopeptidase domain 21 180903 53616 hCG17212.4 ADAM22 ADAM metallopeptidase domain 22 177272 8745 hCG1811623.1 ADAM23 ADAM metallopeptidase domain 23 102384 10863 hCG1818505.1 ADAM28 ADAM metallopeptidase domain 28 119968 11086 hCG1786734.2 ADAM29 ADAM metallopeptidase domain 29 205542 11085 hCG1997196.1 ADAM30 ADAM metallopeptidase domain 30 148417 80332 hCG39255.4 ADAM33 ADAM metallopeptidase domain 33 140492 8756 hCG1789002.2 ADAM7 ADAM metallopeptidase domain 7 122603 101 hCG1816947.1 ADAM8 ADAM metallopeptidase domain 8 183965 8754 hCG1996391 ADAM9 ADAM metallopeptidase domain 9 (meltrin gamma) 129974 27299 hCG15447.3 ADAMDEC1 ADAM-like,
    [Show full text]
  • Cd1b Tetramers Bind T Cell Receptors to Identify a Mycobacterial
    Published August 1, 2011 Brief Definitive Report CD1b tetramers bind T cell receptors to identify a mycobacterial glycolipid- reactive T cell repertoire in humans Anne G. Kasmar,1 Ildiko van Rhijn,1,2 Tan-Yun Cheng,1 Marie Turner,3 Chetan Seshadri,1 Andre Schiefner,4 Ravi C. Kalathur,4 John W. Annand,1 Annemieke de Jong,1 John Shires,5 Luis Leon,1 Michael Brenner,1 Ian A. Wilson,4 John D. Altman,5 and D. Branch Moody1 1Division of Rheumatology, Immunology and Allergy, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115 2Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, 3508 TD Utrecht, Netherlands 3Tuberculosis Treatment Unit, Lemuel Shattuck Hospital, Jamaica Plain, MA 02130 4Department of Molecular Biology and Skaggs Institute for Chemical Biology, the Scripps Research Institute, La Jolla, CA 92037 Downloaded from 5Emory Vaccine Center, Atlanta, GA 30329 Microbial lipids activate T cells by binding directly to CD1 and T cell receptors (TCRs) or by indirect effects on antigen-presenting cells involving induction of lipid autoantigens, CD1 transcription, or cytokine release. To distinguish among direct and indirect mechanisms, we developed fluorescent human CD1b tetramers and measured T cell staining. CD1b tetramer staining of T cells requires glucose monomycolate (GMM) antigens, is specific for TCR jem.rupress.org structure, and is blocked by a recombinant clonotypic TCR comprised of TRAV17 and TRBV4-1, proving that CD1b–glycolipid complexes bind the TCR. GMM-loaded tetramers brightly stain a small subpopulation of blood-derived cells from humans infected with Mycobacterium tuberculosis, providing direct detection of a CD1b-reactive T cell reper- toire.
    [Show full text]
  • Flow Reagents Single Color Antibodies CD Chart
    CD CHART CD N° Alternative Name CD N° Alternative Name CD N° Alternative Name Beckman Coulter Clone Beckman Coulter Clone Beckman Coulter Clone T Cells B Cells Granulocytes NK Cells Macrophages/Monocytes Platelets Erythrocytes Stem Cells Dendritic Cells Endothelial Cells Epithelial Cells T Cells B Cells Granulocytes NK Cells Macrophages/Monocytes Platelets Erythrocytes Stem Cells Dendritic Cells Endothelial Cells Epithelial Cells T Cells B Cells Granulocytes NK Cells Macrophages/Monocytes Platelets Erythrocytes Stem Cells Dendritic Cells Endothelial Cells Epithelial Cells CD1a T6, R4, HTA1 Act p n n p n n S l CD99 MIC2 gene product, E2 p p p CD223 LAG-3 (Lymphocyte activation gene 3) Act n Act p n CD1b R1 Act p n n p n n S CD99R restricted CD99 p p CD224 GGT (γ-glutamyl transferase) p p p p p p CD1c R7, M241 Act S n n p n n S l CD100 SEMA4D (semaphorin 4D) p Low p p p n n CD225 Leu13, interferon induced transmembrane protein 1 (IFITM1). p p p p p CD1d R3 Act S n n Low n n S Intest CD101 V7, P126 Act n p n p n n p CD226 DNAM-1, PTA-1 Act n Act Act Act n p n CD1e R2 n n n n S CD102 ICAM-2 (intercellular adhesion molecule-2) p p n p Folli p CD227 MUC1, mucin 1, episialin, PUM, PEM, EMA, DF3, H23 Act p CD2 T11; Tp50; sheep red blood cell (SRBC) receptor; LFA-2 p S n p n n l CD103 HML-1 (human mucosal lymphocytes antigen 1), integrin aE chain S n n n n n n n l CD228 Melanotransferrin (MT), p97 p p CD3 T3, CD3 complex p n n n n n n n n n l CD104 integrin b4 chain; TSP-1180 n n n n n n n p p CD229 Ly9, T-lymphocyte surface antigen p p n p n
    [Show full text]
  • Human Peripheral Blood Gamma Delta T Cells: Report on a Series of Healthy Caucasian Portuguese Adults and Comprehensive Review of the Literature
    cells Article Human Peripheral Blood Gamma Delta T Cells: Report on a Series of Healthy Caucasian Portuguese Adults and Comprehensive Review of the Literature 1, 2, 1, 1, Sónia Fonseca y, Vanessa Pereira y, Catarina Lau z, Maria dos Anjos Teixeira z, Marika Bini-Antunes 3 and Margarida Lima 1,* 1 Laboratory of Cytometry, Unit for Hematology Diagnosis, Department of Hematology, Hospital de Santo António (HSA), Centro Hospitalar Universitário do Porto (CHUP), Unidade Multidisciplinar de Investigação Biomédica, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto (UMIB/ICBAS/UP), 4099-001 Porto Porto, Portugal; [email protected] (S.F.); [email protected] (C.L.); [email protected] (M.d.A.T.) 2 Department of Clinical Pathology, Centro Hospitalar de Vila Nova de Gaia/Espinho (CHVNG/E), 4434-502 Vila Nova de Gaia, Portugal; [email protected] 3 Laboratory of Immunohematology and Blood Donors Unit, Department of Hematology, Hospital de Santo António (HSA), Centro Hospitalar Universitário do Porto (CHUP), Unidade Multidisciplinar de Investigação Biomédica, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto (UMIB/ICBAS/UP), 4099-001Porto, Portugal; [email protected] * Correspondence: [email protected]; Tel.: + 351-22-20-77-500 These authors contributed equally to this work. y These authors contributed equally to this work. z Received: 10 February 2020; Accepted: 13 March 2020; Published: 16 March 2020 Abstract: Gamma delta T cells (Tc) are divided according to the type of Vδ and Vγ chains they express, with two major γδ Tc subsets being recognized in humans: Vδ2Vγ9 and Vδ1.
    [Show full text]
  • Targeting EZH2 Increases Therapeutic Efficacy of PD-1 Check-Point Blockade in Models of Prostate Cancer Supplement Figures and T
    Targeting EZH2 Increases Therapeutic Efficacy of PD-1 Check-Point Blockade in Models of Prostate Cancer Supplement Figures and Tables 1 Fig. S1. (A) Schema and genotyping PCR example for the creation of EM and EMC genetically engineered mice. (B) Three-dimensional PCa organoids generated from EM mice (without PSACreERT2) alleles. When treated with tamoxifen, demonstrates no loss of H3K27me3 or EDU staining, indicating specificity of tamoxifen-PSACreERT2 mediated deletion of the Ezh2 set domain. (C) Principle component analysis (PCA) following chemical and genetic inhibition of Ezh2 catalytic function results in significant changes in gene expression. 2 Fig. S2. (A) A 29-gene signature derived from Fig. 1C demonstrates complete independence from a previously published polycomb repression signature. (B) Our 29 gene signature demonstrates significant correlation with a previously published polycomb repression signature in 2 independent human PCa gene expression datasets. (C) EZH2 activity is not determined by EZH2 mRNA expression. 3 Fig. S3. A 29-gene signature derived from Fig 1C was used to generate signature scores for each patient within four independent human prostate cancer RNA-seq datasets. Patients were ranked highest score to lowest score and subject to quartile separation. First (blue) and fourth (red) quartiles were analyzed by supervised clustering to demonstrate expression differences within patients with most lowest EZH2 activity and most highest EZH2 activity. 4 Fig. S4. Genes representing IFN signaling (STAT1, IRF9), Th1 chemokines (CXCL10, CXCL11), and MHC Class I molecules (B2M, HLA-A) were shown to be enriched in PCa patients with low EZH2 activity. 5 Fig. S5. Treatment of 22Rv1 human 2D cell lines with the demonstrated conditions for 96 hours show that EZH2 inhibition increases expression of dsRNA (green = dsRNA, blue = nuclei).
    [Show full text]
  • Supplementary Table 1: Differentially Methylated Genes and Functions of the Genes Before/After Treatment with A) Doxorubicin and B) FUMI and in C) Responders Vs
    Supplementary Table 1: Differentially methylated genes and functions of the genes before/after treatment with a) doxorubicin and b) FUMI and in c) responders vs. non- responders for doxorubicin and d) FUMI Differentially methylated genes before/after treatment a. Doxo GENE FUNCTION CCL5, CCL8, CCL15, CCL21, CCR1, CD33, IL5, immunoregulatory and inflammatory processes IL8, IL24, IL26, TNFSF11 CCNA1, CCND2, CDKN2A cell cycle regulators ESR1, FGF2, FGF14, FGF18 growth factors WT1, RASSF5, RASSF6 tumor suppressor b. FUMI GENE FUNCTION CCL7, CCL15, CD28, CD33, CD40, CD69, TNFSF18 immunoregulatory and inflammatory processes CCND2, CDKN2A cell cycle regulators IGF2BP1, IGFBP3 growth factors HOXB4, HOXB6, HOXC8 regulation of cell transcription WT1, RASSF6 tumor suppressor Differentially methylated genes in responders vs. non-responders c. Doxo GENE FUNCTION CBR1, CCL4, CCL8, CCR1, CCR7, CD1A, CD1B, immunoregulatory and inflammatory processes CD1D, CD1E, CD33, CD40, IL5, IL8, IL20, IL22, TLR4 CCNA1, CCND2, CDKN2A cell cycle regulators ESR2, ERBB3, FGF11, FGF12, FGF14, FGF17 growth factors WNT4, WNT16, WNT10A implicated in oncogenesis TNFSF12, TNFSF15 apoptosis FOXL1, FOXL2, FOSL1,HOXA2, HOXA7, HOXA11, HOXA13, HOXB4, HOXB6, HOXB8, HOXB9, HOXC8, regulation of cell transcription HOXD8, HOXD9, HOXD11 GSTP1, MGMT DNA repair APC, WT1 tumor suppressor d. FUMI GENE FUNCTION CCL1, CCL3, CCL5,CCL14, CD1B, CD33, CD40, CD69, immunoregulatory and inflammatory IL20, IL32 processes CCNA1, CCND2, CDKN2A cell cycle regulators IGF2BP1, IGFBP3, IGFBP7, EGFR, ESR2,RARB2
    [Show full text]
  • Discovery of Deoxyceramides and Diacylglycerols As Cd1b Scaffold Lipids Among Diverse Groove-Blocking Lipids of the Human CD1 System
    Discovery of deoxyceramides and diacylglycerols as CD1b scaffold lipids among diverse groove-blocking lipids of the human CD1 system Shouxiong Huanga, Tan-Yun Chenga, David C. Younga, Emilie Layrea, Cressida A. Madigana, John Shiresb, Vincenzo Cerundoloc, John D. Altmanb, and D. Branch Moodya,1 aDepartment of Medicine, Division of Rheumatology, Immunology and Allergy, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115; bEmory Vaccine Center, Emory School of Medicine, Atlanta, GA 30322; and cMedical Research Council Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, United Kingdom Edited* by Peter Cresswell, Yale University School of Medicine, New Haven, CT, and approved October 17, 2011 (received for review August 10, 2011) Unlike the dominant role of one class II invariant chain peptide (CLIP) environment (7, 8). The differing pH requirements for antigen in blocking MHC class II, comparative lipidomics analysis shows that loading, combined with enrichment of endogenous lipids in the human cluster of differentiation (CD) proteins CD1a, CD1b, CD1c, secretory pathway and exogenous lipids endosomes, are coalescing and CD1d bind lipids corresponding to hundreds of diverse accurate into a two-step model of lipid antigen presentation. First, newly mass retention time values. Although most ions were observed in translated CD1 proteins, aided by the microsomal triglyceride association with several CD1 proteins, ligands binding selectively to transport protein (MTP) (9),
    [Show full text]
  • Mouse and Human Fcr Effector Functions
    Pierre Bruhns Mouse and human FcR effector € Friederike Jonsson functions Authors’ addresses Summary: Mouse and human FcRs have been a major focus of Pierre Bruhns1,2, Friederike J€onsson1,2 attention not only of the scientific community, through the cloning 1Unite des Anticorps en Therapie et Pathologie, and characterization of novel receptors, and of the medical commu- Departement d’Immunologie, Institut Pasteur, Paris, nity, through the identification of polymorphisms and linkage to France. disease but also of the pharmaceutical community, through the iden- 2INSERM, U760, Paris, France. tification of FcRs as targets for therapy or engineering of Fc domains for the generation of enhanced therapeutic antibodies. The Correspondence to: availability of knockout mouse lines for every single mouse FcR, of Pierre Bruhns multiple or cell-specific—‘a la carte’—FcR knockouts and the Unite des Anticorps en Therapie et Pathologie increasing generation of hFcR transgenics enable powerful in vivo Departement d’Immunologie approaches for the study of mouse and human FcR biology. Institut Pasteur This review will present the landscape of the current FcR family, 25 rue du Docteur Roux their effector functions and the in vivo models at hand to study 75015 Paris, France them. These in vivo models were recently instrumental in re-defining Tel.: +33145688629 the properties and effector functions of FcRs that had been over- e-mail: [email protected] looked or discarded from previous analyses. A particular focus will be made on the (mis)concepts on the role of high-affinity Acknowledgements IgG receptors in vivo and on results from antibody engineering We thank our colleagues for advice: Ulrich Blank & Renato to enhance or abrogate antibody effector functions mediated by Monteiro (FacultedeMedecine Site X.
    [Show full text]