DOCSLIB.ORG

University of Southampton Research Repository Eprints Soton

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
University of Southampton Research Repository Eprints Soton University of Southampton Research Repository ePrints Soton Copyright © and Moral Rights for this thesis are retained by the author and/or other copyright owners. A copy can be downloaded for personal non-commercial research or study, without prior permission or charge. This thesis cannot be reproduced or quoted extensively from without first obtaining permission in writing from the copyright holder/s. The content must not be changed in any way or sold commercially in any format or medium without the formal permission of the copyright holders. When referring to this work, full bibliographic details including the author, title, awarding institution and date of the thesis must be given e.g. AUTHOR (year of submission) "Full thesis title", University of Southampton, name of the University School or Department, PhD Thesis, pagination http://eprints.soton.ac.uk UNIVERSITY OF SOUTHAMPTON FACULTY OF MEDICINE School of Medicine Department of Cancer Sciences Manipulation of immunotherapy by targeting the inhibitory Fcγ receptor. by Emily Louisa Williams, BSc. (Hons) Thesis for the degree of Doctor of Philosophy June 2011 2 UNIVERSITY OF SOUTHAMPTON ABSTRACT FACULTY OF MEDICINE, SCHOOL OF MEDICINE, DEPARTMENT OF CANCER SCIENCES Doctor of Philosophy Manipulation of immunotherapy by targeting the inhibitory Fcγ receptor. Emily Louisa Williams BSc. (Hons) The inhibitory Fc receptor for IgG, FcγRII (CD32), has been shown to modulate in vivo cytotoxicity against tumour targets. Usually expressed on monocytes, macrophages and B lymphocytes, CD32 deficiency enhances the efficacy of some therapeutic mAb. Unfortunately, suitable reagents capable of specifically and exclusively binding mouse CD32 have been lacking, leading to an ignorance of how CD32 might be manipulated for therapeutic benefit in vivo. We generated a panel of mAb which are highly specific for CD32 that do not cross-react with any other Fc receptors, allowing us to study the potential of CD32 as a therapeutic target able to alter effector cell function and concurrent immunotherapy. In vitro analysis on murine lymphoma cell lines showed that the anti-CD32 mAb fall into two distinct categories; those which cause tyrosine phosphorylation and subsequent receptor activation (agonistic) and those which block receptor phosphorylation (antagonistic). Consequentially, these anti-CD32 mAb exert differing functions in a range of cellular and biochemical assays. Agonistic mAb trigger programmed cell death whilst antagonistic anti-CD32 mAb have relatively little effect. Interestingly, the agonistic mAb are able to block the magnitude and duration of the calcium signalling triggered by BCR engagement whilst antagonistic mAb potentiate these responses. Furthermore, these mAb were shown to trigger phagocytosis of antibody coated B-cells by murine bone marrow derived macrophages (BMDMs) and to enhance the phagocytic potential of BMDMs in vitro. The ability of these mAb to elicit therapeutic responses in a variety of different mouse models was however limited. Further study revealed that in wild-type mice the anti-CD32 mAb are rapidly consumed in comparison with other, similar, mAb. Together with our observation that the anti-CD32 mAb are internalised in vitro and in vivo, these data provide the rationale for the inadequate immunotherapy observed in our model systems. i ii Table of contents Abstract i Table of contents iii List of figures ix Lists of tables xii Declaration of Authourship xiii Dedication xv Acknowledgements xvii Abbreviations xix CHAPTER 1 Introduction 1 1.1. Introduction. 1 1.2. Immunotherapy. 2 1.2.1. Active immunotherapeutic strategies. 3 1.2.1.1. Cancer vaccination. 3 1.2.1.2. Immunostimulatory mAb. 5 1.2.2. Passive immunotherapeutic strategies. 6 1.3. Direct targeting mAb immunotherapy. 7 1.3.1. Antibody structure. 7 1.3.2. Development of mAb technology. 9 1.3.3. Effector mechanism of therapeutic mAb. 11 1.3.3.1. Induction of ADCC. 12 1.3.3.2. Induction of CDC. 12 1.3.3.3. Signal perturbation (PCD and altered endogenous signalling). 13 1.3.3.4. Additional anti-tumour functions. 15 1.3.4. Improving therapeutic mAb efficacy. 17 1.3.4.1. mAb conjugation. 17 1.3.4.2. Maximising FcγR:Fc interactions. 18 1.4. The Fc receptor family. 19 1.5. The Fcγ Receptors. 19 1.5.1. FcγR binding affinity for IgG. 21 1.5.2. The activatory-to-inhibitory ratio and its importance in cell activation. 22 1.5.3. Structure of the FcγR. 23 1.5.4. The FcγR:Fc interaction (binding of IgG to FcγR). 25 1.5.5. Regulation of FcγR expression. 27 1.6. The activatory FcγR. 28 1.6.1. The function of activatory FcγR. 28 1.6.2. Activatory FcγR signalling. 29 1.7. B cell signalling. 32 1.7.1. BCR-induced cytosolic calcium flux. 33 1.7.2. BCR-induced activation of MAP kinase signalling pathway. 34 1.7.3. BCR-induced activation of AKT. 34 1.7.4. The important of lipid rafts in ITAM receptor signalling. 35 iii 1.8. The inhibitory FcγR. 37 1.8.1. Expression of CD32 38 1.8.1.1. Expression of CD32 isoforms. 38 1.8.2. Inhibition of activatory FcγR signalling. 40 1.8.3. Inhibition of BCR signalling. 41 1.8.3.1. CD32 recruitment to lipid rafts after BCR co-ligation. 42 1.8.3.2. Inhibition of BCR-induced cytosolic calcium flux. 43 1.8.3.3. Decreased survival by inhibition of AKT. 43 1.8.3.4. Reduced cell proliferation by inhibition of MAP kinase pathways. 44 1.8.4. Direct B-cell apoptosis induced by CD32 activation. 46 1.9. Expression of FcγR in cancer. 48 1.10. FcγR as mediators of anti-tumour immunotherapy. 50 1.10.1. The role of FcγR in immunotherapy. 50 1.10.2. Inhibitory FcγR in immunotherapy. 50 1.11. Project aims. 52 CHAPTER 2 Materials and Methods 53 2.1. Animals. 53 2.2. Tissue culture. 53 2.3. Cell quantification. 54 2.4. B cell purification by MACS® separation. 54 2.5. Lymphoprep of whole blood. 55 2.6. Mouse genotyping by polymerase chain reaction (PCR). 55 2.6.1. Myc genotyping. 56 2.6.2. CD32-/- genotyping. 56 2.6.3. γ-chain-/- genotyping. 56 2.6.4. Bim-/- genotyping. 57 2.7. Mouse phenotyping by immunofluroscence. 57 2.8. Eµ-myc mice; tumour generation, processing and characterisation. 58 2.9. Antibodies and antibody fragments. 58 2.9.1. Generation of antibodies. 58 2.9.2. Generation of F(ab’)2 antibody fragments. 59 2.10. Antibody dialysis. 59 2.11. Measurement of surface antigens by immunofluorescence. 59 2.12. In vitro Apoptosis Assay. 60 2.12.1. AnV/PI staining of lymphoma and B cells. 60 2.12.2. AnV/PI staining of BMDMs. 61 2.13. Calcium flux assay. 61 2.14. Western blot analysis. 62 iv 2.15. mAb immunotherapy. 63 2.16. Tumour tracking. 64 2.17. Ex vivo modulation assay. 64 2.18. In vitro Modulation assays. 64 2.19. Alexa-488 quenching Assay. 65 2.19.1. Alexa-488 labelling of mAb. 65 2.19.2. Quenching assay. 65 2.20. Adoptive transfer of CFSE labelled cells. 66 2.20.1. CFSE labelling of target and non-target splenocytes. 66 2.20.2. Adoptive transfer of target and non-target cells. 66 2.21. BMDMs phagocytosis assay. 66 2.22. ELISA to determine half life in vivo of anti-CD32 mAb. 67 2.23. Production of clodronate liposomes. 68 2.24. Polymerase chain reaction for murine CD32 isoforms. 68 2.24.1. RNA extraction. 69 2.24.2. cDNA production. 69 2.24.3. PCR for murine CD32 isoforms. 69 2.25. BIAcore analysis to determine affinity of anti-CD32 mAb. 69 2.26. Production of immune complexes. 70 CHAPTER 3 In vitro characterisation of the Eµ-myc mouse model 73 3.1. Chapter introduction. 73 3.2. Kaplein Meier survival. 75 3.3. Presentation of the primary tumour. 76 3.3.1. Physical tumour presentation. 76 3.3.2. Histological examination of tissues from tumour bearing mice. 80 3.3.2.1. Histological examination of lymphoid tissue from the C57BL/6 and Eµ-myc mice. 80 3.3.2.2. Histological examination of non-lymphoid tissue from C57BL/6 and Eµ-myc mice. 81 3.3.2.3. Histological examination of tissues from the Eµ-myc/CD32-/- mice. 81 3.4. Primary tumour characterisation. 86 3.4.1. Tumour phenotype. 86 3.4.2. Tumour transplantation into WT C57BL/6 mice. 89 3.5. Eμ-myc and Eµ-myc/CD32-/- cell line characterisation 90 3.5.1. Eµ-myc and Eµ-myc/CD32-/- cell line growth 90 3.5.2. Changes in IgM expression as cells adapt to culture. 91 3.5.3. Response of Eµ-myc cell-lines to etoposide or BCR stimulation 92 3.6. Chapter Discussion 97 v CHAPTER 4 In vitro analysis of anti-CD32 mAb on lymphoma cells 103 4.1. Chapter Introduction. 103 4.2. Generation of anti-CD32 mAb. 103 4.2.1. anti-CD32 mAb binding specificity and affinity. 104 4.2.2. Identification of epitope specificity. 108 4.2.3. The anti-CD32 mAb block immune complex binding. 108 4.3. Activation of CD32. 111 4.3.1. Tyrosine phosphorylation of CD32, SHIP-1 and SHP by anti-BCR antibodies. 111 4.3.2. Tyrosine phosphorylation of CD32 by the anti-CD32 mAb. 112 4.3.3. Tyrosine phosphorylation of SHIP-1 following activation of CD32 by the anti- CD32 mAb. 113 4.3.4. Tyrosine phosphorylation of CD32 by the anti-CD32 mAb whole IgG and F(ab’)2 fragments. 115 4.3.5. Co-ligation of CD32 by AT130-2 and AT130-5 is in a cis fashion.
Load more

Recommended publications
  • The Ligands for Human Igg and Their Effector Functions
    antibodies Review The Ligands for Human IgG and Their Effector Functions Steven W. de Taeye 1,2,*, Theo Rispens 1 and Gestur Vidarsson 2 1 Sanquin Research, Dept Immunopathology and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, 1066 CX Amsterdam, The Netherlands; [email protected] 2 Sanquin Research, Dept Experimental Immunohematology and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, 1066 CX Amsterdam, The Netherlands; [email protected] * Correspondence: [email protected] Received: 26 March 2019; Accepted: 18 April 2019; Published: 25 April 2019 Abstract: Activation of the humoral immune system is initiated when antibodies recognize an antigen and trigger effector functions through the interaction with Fc engaging molecules. The most abundant immunoglobulin isotype in serum is Immunoglobulin G (IgG), which is involved in many humoral immune responses, strongly interacting with effector molecules. The IgG subclass, allotype, and glycosylation pattern, among other factors, determine the interaction strength of the IgG-Fc domain with these Fc engaging molecules, and thereby the potential strength of their effector potential. The molecules responsible for the effector phase include the classical IgG-Fc receptors (FcγR), the neonatal Fc-receptor (FcRn), the Tripartite motif-containing protein 21 (TRIM21), the first component of the classical complement cascade (C1), and possibly, the Fc-receptor-like receptors (FcRL4/5). Here we provide an overview of the interactions of IgG with effector molecules and discuss how natural variation on the antibody and effector molecule side shapes the biological activities of antibodies. The increasing knowledge on the Fc-mediated effector functions of antibodies drives the development of better therapeutic antibodies for cancer immunotherapy or treatment of autoimmune diseases.
    [Show full text]
  • Pharmacologic Considerations in the Disposition of Antibodies and Antibody-Drug Conjugates in Preclinical Models and in Patients
    antibodies Review Pharmacologic Considerations in the Disposition of Antibodies and Antibody-Drug Conjugates in Preclinical Models and in Patients Andrew T. Lucas 1,2,3,*, Ryan Robinson 3, Allison N. Schorzman 2, Joseph A. Piscitelli 1, Juan F. Razo 1 and William C. Zamboni 1,2,3 1 University of North Carolina (UNC), Eshelman School of Pharmacy, Chapel Hill, NC 27599, USA; [email protected] (J.A.P.); [email protected] (J.F.R.); [email protected] (W.C.Z.) 2 Division of Pharmacotherapy and Experimental Therapeutics, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; [email protected] 3 Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; [email protected] * Correspondence: [email protected]; Tel.: +1-919-966-5242; Fax: +1-919-966-5863 Received: 30 November 2018; Accepted: 22 December 2018; Published: 1 January 2019 Abstract: The rapid advancement in the development of therapeutic proteins, including monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs), has created a novel mechanism to selectively deliver highly potent cytotoxic agents in the treatment of cancer. These agents provide numerous benefits compared to traditional small molecule drugs, though their clinical use still requires optimization. The pharmacology of mAbs/ADCs is complex and because ADCs are comprised of multiple components, individual agent characteristics and patient variables can affect their disposition. To further improve the clinical use and rational development of these agents, it is imperative to comprehend the complex mechanisms employed by antibody-based agents in traversing numerous biological barriers and how agent/patient factors affect tumor delivery, toxicities, efficacy, and ultimately, biodistribution.
    [Show full text]
  • Transfer of Igg in the Female Genital Tract by MHC Class I-Related Neonatal Fc Receptor (Fcrn) Confers Protective Immunity to Vaginal Infection
    Transfer of IgG in the female genital tract by MHC class I-related neonatal Fc receptor (FcRn) confers protective immunity to vaginal infection Zili Lia,b, Senthilkumar Palaniyandia,b, Rongyu Zenga,b, Wenbin Tuoc, Derry C. Roopeniand, and Xiaoping Zhua,b,1 aLaboratory of Immunology, Virginia-Maryland Regional College of Veterinary Medicine, College Park, MD 20742; bMaryland Pathogen Research Institute, University of Maryland, College Park, MD 20742; cAnimal Parasitic Diseases Laboratory, Agricultural Research Service, United States Department of Agriculture, Beltsville, MD 20705; and dThe Jackson Laboratory, Bar Harbor, ME 04609 Edited by Roy Curtiss, Arizona State University, Tempe, AZ, and approved February 1, 2011 (received for review August 30, 2010) IgG is a major Ig subclass in mucosal secretions of the human and viral load. Therefore, HIV-specific IgG may be much more female genital tract, where it predominates over the IgA isotype. important in mucosal protection than previously thought. Unlike Despite the abundance of IgG, surprisingly little is known about S-IgA, the mechanism(s) by which the IgG antibody is transported where and how IgG enters the lumen of the genital tract and the across the genital epithelium and the role of IgG in genital mu- exact role local IgG plays in preventing sexually transmitted dis- cosal protection have not been investigated. Incomplete un- eases. We demonstrate here that the neonatal Fc receptor, FcRn, is derstanding of IgG transport in the genital tract and of its role in expressed in female genital tract epithelial cells of humans and mice combating genital infections has hampered the design and de- and binds IgG in a pH-dependent manner.
    [Show full text]
  • Alpha-Dystroglycan Plays Functional Roles in Platelet Aggregation and Thrombus Growth
    Alpha-dystroglycan plays functional roles in platelet aggregation and thrombus growth by Reid Gallant A thesis submitted in conformity with the requirements For the degree of Master of Science Graduate Department of Laboratory Medicine and Pathobiology University of Toronto © Copyright by Reid Gallant 2017 i Alpha-dystroglycan Plays Functional Roles in Platelet Aggregation and Thrombus Growth Reid Gallant Master of Science Department of Laboratory Medicine and Pathobiology University of Toronto 2017 ABSTRACT Fibrinogen (Fg) and von Willebrand factor (VWF) have been considered essential for platelet adhesion and aggregation. However, platelet aggregation still occurs in mice lacking Fg and/or VWF but not β3 integrin, suggesting other, unidentified αIIbβ3 integrin ligand(s) mediate platelet aggregation. Through screening published platelet proteomics data, we identified a candidate, alpha-dystroglycan (α-DG). Using Western blot and flow cytometry, I found α-DG is expressed on platelets. Using aggregometry, I observed that antibodies against α-DG or its N- terminal Laminin-binding site, decreased platelet aggregation induced by various platelet agonists in both platelet-rich plasma and gel-filtered platelets. These antibodies also decreased platelet adhesion/aggregation in perfusion chambers independent of α-DG-Laminin interaction. Using laser injury intravital microscopy and carotid artery thrombosis models, we further found that these anti-α-DG antibodies decreased thrombus growth in vivo. Our results showed that α- DG may form an α-DG-fibronectin complex that binds to αIIbβ3 integrin, contributing to platelet adhesion/aggregation, and thrombosis growth. ii Acknowledgements ―It helps a man immensely to be a bit of a hero-worshipper, and the stories of the lives of the masters of medicine do much to stimulate our ambition and rouse our sympathies‖ – Sir William Osler I will always be grateful to my MSc supervisor, Dr.
    [Show full text]
  • Ncounter® Mouse Autoimmune Profiling Panel - Gene and Probe Details
    nCounter® Mouse AutoImmune Profiling Panel - Gene and Probe Details Official Symbol Accession Alias / Previous Symbol Official Full Name Other targets or Isoform Information AW208573,CD143,expressed sequence AW208573,MGD-MRK- Ace NM_009598.1 1032,MGI:2144508 angiotensin I converting enzyme (peptidyl-dipeptidase A) 1 2610036I19Rik,2610510L13Rik,Acinus,apoptotic chromatin condensation inducer in the nucleus,C79325,expressed sequence C79325,MGI:1913562,MGI:1919776,MGI:2145862,mKIAA0670,RIKEN cDNA Acin1 NM_001085472.2 2610036I19 gene,RIKEN cDNA 2610510L13 gene apoptotic chromatin condensation inducer 1 Acp5 NM_001102405.1 MGD-MRK-1052,TRACP,TRAP acid phosphatase 5, tartrate resistant 2310066K23Rik,AA960180,AI851923,Arp1b,expressed sequence AA960180,expressed sequence AI851923,MGI:2138136,MGI:2138359,RIKEN Actr1b NM_146107.2 cDNA 2310066K23 gene ARP1 actin-related protein 1B, centractin beta Adam17 NM_001277266.1 CD156b,Tace,tumor necrosis factor-alpha converting enzyme a disintegrin and metallopeptidase domain 17 ADAR1,Adar1p110,Adar1p150,AV242451,expressed sequence Adar NM_001038587.3 AV242451,MGI:2139942,mZaADAR adenosine deaminase, RNA-specific Adora2a NM_009630.2 A2AAR,A2aR,A2a, Rs,AA2AR,MGD-MRK-16163 adenosine A2a receptor Ager NM_007425.2 RAGE advanced glycosylation end product-specific receptor AI265500,angiotensin precursor,Aogen,expressed sequence AI265500,MGD- Agt NM_007428.3 MRK-1192,MGI:2142488,Serpina8 angiotensinogen (serpin peptidase inhibitor, clade A, member 8) Ah,Ahh,Ahre,aromatic hydrocarbon responsiveness,aryl hydrocarbon
    [Show full text]
  • I-Related Neonatal Fc Receptor for Igg This Information Is Current As of September 26, 2021
    Activation of the JAK/STAT-1 Signaling Pathway by IFN- γ Can Down-Regulate Functional Expression of the MHC Class I-Related Neonatal Fc Receptor for IgG This information is current as of September 26, 2021. Xindong Liu, Lilin Ye, Yu Bai, Habi Mojidi, Neil E. Simister and Xiaoping Zhu J Immunol 2008; 181:449-463; ; doi: 10.4049/jimmunol.181.1.449 http://www.jimmunol.org/content/181/1/449 Downloaded from References This article cites 74 articles, 35 of which you can access for free at: http://www.jimmunol.org/content/181/1/449.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 26, 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 Activation of the JAK/STAT-1 Signaling Pathway by IFN-␥ Can Down-Regulate Functional Expression of the MHC Class I-Related Neonatal Fc Receptor for IgG1 Xindong Liu,* Lilin Ye,* Yu Bai,* Habi Mojidi,*† Neil E.
    [Show full text]
  • Identification and Characterization of Fcγrs in Göttingen Minipigs – Implications for Preclinical Assessment of Therapeutic Antibodies
    Identification and characterization of FcγRs in Göttingen minipigs – implications for preclinical assessment of therapeutic antibodies Inauguraldissertation zur Erlangung der Würde eines Doktors der Philosophie vorgelegt der Philosophisch-Naturwissenschaftlichen Fakultät der Universität Basel von Jérôme Egli aus Neckertal (SG), Schweiz Basel, 2019 Originaldokument gespeichert auf dem Dokumentenserver der Universität Basel edoc.unibas.ch Genehmigt von der Philosophisch-Naturwissenschaftlichen Fakultät auf Antrag von Prof. Dr. Alex Odermatt (Fakultätsverantwortlicher), PD Dr. Antonio Iglesias (Dissertationsleiter), und Prof. Dr. Daniela Finke (Korreferentin) Basel, den 11.12.2018 ______________________ Dekan Prof. Dr. Martin Spiess Table of contents 1 Abstract ................................................................................................................................ 1 2 Abbreviations ....................................................................................................................... 3 3 Preface ................................................................................................................................. 5 4 Introduction.......................................................................................................................... 6 4.1 Therapeutic antibodies and effector functions ....................................................................... 6 4.2 Preclinical studies with therapeutic antibodies ...................................................................
    [Show full text]
  • Characterization and Localization of the Neonatal Fc Receptor in Adult Human Kidney
    J Am Soc Nephrol 11: 632–639, 2000 Characterization and Localization of the Neonatal Fc Receptor in Adult Human Kidney JEAN-PHILIPPE HAYMANN,* JEAN-PIERRE LEVRAUD,† SANDRINE BOUET,* VINCENT KAPPES,* JACQUELINE HAGEGE,*` GENEVIEVE` NGUYEN,* YICHUN XU,* ERIC RONDEAU,* and JEAN-DANIEL SRAER* *Service de Ne´phrologie A, Assistance Publique-Hoˆpitaux de Paris, Institut National de la Sante´etdela Recherche Me´dicale U489 et Association Claude Bernard, Hoˆpital Tenon, and † Institut National de la Sante´ et de la Recherche Me´dicale U277, Institut Pasteur, Paris, France. Abstract. The binding of Fc fragments of Ig on glomerular by flow cytometry, reverse transcription-PCR, Western blot- epithelial cells (GEC) was observed previously, but the recep- ting, and by the pH dependence of the binding of heat-aggre- tor could not be identified. In immunofluorescence and immu- gated IgG. Because it is well established that the FcRn is nohistochemical studies using normal adult human kidney sec- involved in IgG transcytosis, it is hypothesized that the FcRn in tions, the presence of the so-called neonatal Fc receptor (FcRn) the kidney may play a role in the reabsorption of IgG. Ongoing was demonstrated on GEC as well as in the brush border of studies should clarify the role of the FcRn as a potential target proximal tubular cells. FcRn transcripts were also detected on for immune complexes on GEC and should assess its relevance isolated glomeruli by reverse transcription-PCR. Using an im- in physiology and pathology. mortalized GEC line, the presence of the FcRn was confirmed Immune complex deposition is encountered in many glomer- transcribed in many adult tissues (7,8,19), has been identified ulonephritides.
    [Show full text]
  • Defining the Gut-Mammary Gland-Secretory Iga Axis in Porcine Epidemic Diarrhea Virus Infected Gilts and Its Impact on Lactogenic
    Defining the Gut-Mammary Gland-Secretory IgA Axis in Porcine Epidemic Diarrhea Virus Infected Gilts and its Impact on Lactogenic Immune Protection of Neonatal Suckling Piglets Dissertation Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Stephanie Langel Graduate Program in Comparative and Veterinary Medicine The Ohio State University 2018 Dissertation Committee Dr. Linda J. Saif, Advisor and Committee Chair Dr. Anastasia N. Vlasova, Committee Member Dr. Qiuhong Wang, Committee Member Dr. Prosper N. Boyaka, Committee Member Copyrighted by Stephanie Mary Neal Langel 2018 2 Abstract Neonates are immune competent but immunologically immature at birth and are highly susceptible to enteric pathogens. For example, porcine epidemic diarrhea virus (PEDV) is a highly virulent re-emerging enteric coronavirus that causes acute diarrhea, dehydration and 80-100% mortality in neonatal piglets. Since its emergence in the United States (US) in 2013, PEDV caused over 8.5 million piglet deaths and $900 million to $1.8 billion in losses to US swine producers in 2014. Lactogenic immunity remains the most promising and effective way to protect neonatal suckling piglets from enteric diseases like PEDV. This is particularly true for domesticated animals like swine and cattle whose epitheliochorial placenta inhibits immunoglobulin (Ig) transfer in utero. Therefore, colostrum [first secretions from the mammary gland (MG)] and milk-derived antibodies (Abs) and other immune factors are the sole source for immune protection after birth. Passive lactogenic immunity is achieved through high titers of IgG Abs in colostrum and a continuous supply of secretory IgA (sIgA) Abs in milk.
    [Show full text]
  • The Neonatal Fc Receptor and Complement Fixation Facilitate
    The Neonatal Fc Receptor and Complement Fixation Facilitate Prophylactic Vaccine-Mediated Humoral Protection against Viral Infection in the Ocular Mucosa This information is current as of October 2, 2021. Derek J. Royer, Meghan M. Carr, Hem R. Gurung, William P. Halford and Daniel J. J. Carr J Immunol published online 31 July 2017 http://www.jimmunol.org/content/early/2017/07/29/jimmun ol.1700316 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2017/07/29/jimmunol.170031 Material 6.DCSupplemental http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on October 2, 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 © 2017 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published July 31, 2017, doi:10.4049/jimmunol.1700316 The Journal of Immunology The Neonatal Fc Receptor and Complement Fixation Facilitate Prophylactic Vaccine-Mediated Humoral Protection against Viral Infection in the Ocular Mucosa Derek J. Royer,* Meghan M. Carr,* Hem R. Gurung,† William P.
    [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]
  • Activation of the JNK/MAPK Signaling Pathway by TGF-1 Enhances Neonatal Fc Receptor Expression and Igg Transcytosis
    microorganisms Communication Activation of the JNK/MAPK Signaling Pathway by TGF-β1 Enhances Neonatal Fc Receptor Expression and IgG Transcytosis Shaoju Qian 1, Chenxi Li 1, Xi Liu 1, Xiangchao Jia 1, Yuncai Xiao 1,2 and Zili Li 1,2,3,* 1 State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; [email protected] (S.Q.); [email protected] (C.L.); [email protected] (X.L.); [email protected] (X.J.); [email protected] (Y.X.) 2 Key Laboratory of Preventive Veterinary Medicine in Hubei Province, The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China 3 Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture of the People’s Republic of China, Wuhan 430070, China * Correspondence: [email protected]; Tel.: +86-027-87288917 Abstract: The neonatal Fc receptor (FcRn) transports maternal immunoglobulin G (IgG) to the foetus or newborn and protects the IgG from degradation. FcRn is expressed in several porcine tissues and cell types and its expression levels are regulated by immune and inflammatory events. IPEC-J2 cells are porcine intestinal columnar epithelial cells that were isolated from neonatal piglet mid-jejunum. We hypothesized that transforming growth factor β1 (TGF-β1) upregulated pFcRn expression in IPEC-J2 cells. To test this hypothesis, we treated IPEC-J2 cells with TGF-β1 and demonstrated that porcine FcRn (pFcRn) expression was significantly increased. SP600125, a specific mitogen-activated protein kinase (MAPK) inhibitor, reduced TGF-β1-induced pFcRn expression in IPEC-J2 cells.
    [Show full text]