DOCSLIB.ORG

Development and Assessment of Novel Vaccination Strategies for Hepatitis B Virus Infection

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Development and Assessment of Novel Vaccination Strategies for Hepatitis B Virus Infection DEVELOPMENT AND ASSESSMENT OF NOVEL VACCINATION STRATEGIES FOR HEPATITIS B VIRUS INFECTION Devin TEOH (Chee Quin TEOH) B.Sc. (Biotechnology), B.Sc. (Hons) A thesis submitted to the University of Adelaide in fulfilment of the requirements for the degree of Doctor of Philosophy Discipline of Microbiology and Immunology School of Molecular and Biomedical Science The University of Adelaide South Australia Australia November 2014 To my loving family for your boundless love, care, support and always being there for me & To Jean Ang who stood by me when things looked bleak and supported me each step of the way Table of Contents Abstract-----------------------------------------------------------------------------------------------------i Declaration of originality and consent---------------------------------------------------------------v Publications and presentations resulting from this thesis---------------------------------------vi Acknowledgements--------------------------------------------------------------------------------------ix Abbreviations--------------------------------------------------------------------------------------------xi Chapter 1 Introduction 1.1 Background: Epidemiology of HBV infection--------------------------------------1 1.2 Hepadnaviruses--------------------------------------------------------------------------2 1.2.1 Hepadnavirus Family--------------------------------------------------------------------2 1.2.2 B virus structure and genome organisation-------------------------------------------2 1.2.3 Duck hepatitis B virus structure and genome organisation------------------------9 1.3 Hepadnavirus Replication-------------------------------------------------------------10 1.3.1 Viral entry, transcription and translation to generate new virus-----------------10 1.3.2 An important reservoir of hepadnavirus infection – cccDNA--------------------13 1.4 Outcomes of HBV infection----------------------------------------------------------13 1.4.1 Acute HBV infection------------------------------------------------------------------17 1.4.1.1 Resolution of acute HBV infection--------------------------------------------------17 1.4.2 Chronic HBV infection----------------------------------------------------------------20 1.5 Animal Model – Duck hepatitis B virus--------------------------------------------23 1.6 Outcomes of DHBV infection -------------------------------------------------------23 1.6.1 Acute DHBV infection ---------------------------------------------------------------25 1.6.2 Persistent DHBV infection------------------------------------------------------------25 1.6.3 CMI responses to DHBV infection -------------------------------------------------25 1.7 Treatment of HBV infection ---------------------------------------------------------27 1.7.1 Immunomodulators--------------------------------------------------------------------27 1.7.2 Antiviral agents-------------------------------------------------------------------------28 1.7.2.1 Entecavir--------------------------------------------------------------------------------29 1.7.2.2 Antiviral resistance to Nucleotide/nucleoside analogues-------------------------30 1.8 Hepadnavirus vaccines----------------------------------------------------------------31 1.8.1 The prophylactic HBV vaccines-----------------------------------------------------31 1.8.2 DNA vaccines--------------------------------------------------------------------------32 1.8.2.1 Mechanism of antigen presentation following DNA vaccination---------------34 1.8.2.2 Enhancement of immunogenicity of DNA vaccines------------------------------35 1.8.2.3 Costimulatory molecules as genetic adjuvants ------------------------------------36 1.8.2.4 The co-stimulatory molecule CD40L------------------------------------------------38 1.8.2.5 Current hepadnaviral vaccine studies------------------------------------------------42 1.9 Poxviruses ------------------------------------------------------------------------------43 1.9.1 Family Poxviridae----------------------------------------------------------------------43 1.9.2 Structure and genome of Avipoxviruses--------------------------------------------43 1.9.3 Replication of Avipoxviruses---------------------------------------------------------44 1.9.4 FPV as vaccine vectors----------------------------------------------------------------45 1.9.5 Poxvirus vaccine developed to treat hepadnavirus infection---------------------46 1.10 Research aims and study outline-----------------------------------------------------47 Chapter 2 Materials and Methods 2.1 Experimental animals------------------------------------------------------------------50 2.2 Cell culture------------------------------------------------------------------------------50 2.2.1 Preparation of primary duck embryonic fibroblasts (PDEF)---------------------50 2.2.2 Collection and purification of duck peripheral blood mononuclear cells (PBMC)----------------------------------------------------------------------------------51 2.2.3 Cell culture and PHA and ConA stimulation of spleen cells---------------------51 2.2.4 Culture of 293T, LMH and PDEF cells---------------------------------------------53 2.2.5 Cell transfection------------------------------------------------------------------------53 2.2.6 Detection of DHBV antigens using Immuno-fluorescence (IMF) assay-------54 2.2.7 Detection of DHBV antigens using western blot----------------------------------55 2.3 Production of polyclonal and mAB to recombinant DHBV core antigen (rDHBcAg)------------------------------------------------------------------------------60 2.3.1 Preparation of rDHBcAg particles---------------------------------------------------60 2.3.1.1 Growth and transformed E.coli strains and partial purification of rDHBcAg particles----------------------------------------------------------------------------------60 2.3.1.2 Separation and isolation of rDHBcAg particles by caesium chloride isopycnic density gradient centrifugation-------------------------------------------------------61 2.3.1.3 Detection of rDHBcAg particles by ELISA----------------------------------------62 2.3.1.4 Precipitation of rDHBcAg using trichloroacetic acid (TCA)--------------------63 2.3.1.5 To concentrate rDHBcAg particles and to determine the concentration of rDHBcAg particles --------------------------------------------------------------------63 2.3.2 Production of polyclonal rabbit antibodies to rDHBcAg-------------------------64 2.3.2.1 Testing antibody responses by ELISA following immunisation with rDHBcAg--------------------------------------------------------------------------------64 2.3.2.2 Absorption of polyclonal antisera with normal duck liver (NDL) and NDS---65 2.3.3 Production of mABs to rDHBcAg---------------------------------------------------65 2.3.3.1 Hybridoma production ----------------------------------------------------------------66 2.3.3.2 Hybridoma screening for anti-DHBc antibodies ----------------------------------69 2.4 Procedure involving Duck CD40L (DuCD40L) ----------------------------------69 2.4.1 Sub cloning of DuCD40L gene into pcSNA3 expression vector----------------69 2.4.1.1 Plasmid vector and primer design----------------------------------------------------69 2.4.1.2 RNA extraction-------------------------------------------------------------------------70 2.4.1.3 The cDNA synthesis and cloning by PCR------------------------------------------70 2.4.1.4 Restriction Endonuclease (RE) digestion-------------------------------------------72 2.4.1.5 Ligation----------------------------------------------------------------------------------72 2.4.1.6 Transformation-------------------------------------------------------------------------73 2.4.1.7 Preparation of plasmid DNA---------------------------------------------------------73 2.4.2 Nitrate assay----------------------------------------------------------------------------74 2.4.3 Statistical analysis----------------------------------------------------------------------75 2.4.4 Quantitative reverse transcription PCR (qRT-PCR) to detect iNOS mRNA---75 2.5 Procedures involving ducks-----------------------------------------------------------75 2.5.1 DHBV stock and intravenous inoculation of ducks-------------------------------75 2.5.2 DHBV vaccine and vaccination protocol-------------------------------------------76 2.5.2.1 DNA vaccine----------------------------------------------------------------------------76 2.5.2.2 Vaccination protocols------------------------------------------------------------------77 2.5.3 Administration of antiviral drugs----------------------------------------------------77 2.5.4 Venipuncture----------------------------------------------------------------------------78 2.5.5 Liver Biopsy----------------------------------------------------------------------------78 2.5.6 Autopsy----------------------------------------------------------------------------------79 2.5.7 Tissue fixation, embedding and sectioning-----------------------------------------79 2.6 Serological Assays --------------------------------------------------------------------79 2.6.1 Quantitative detection of DHBsAg by ELISA-------------------------------------80 2.6.2 Quantitative detection of anti-DHBs antibodies by ELISA --------------------- 80 2.6.3 Detection of anti-DHBc antibodies by ELISA-------------------------------------81 2.7 Histology and immuno-staining of tissue sections--------------------------------82 2.7.1 Immuno-staining to detect DHBsAg------------------------------------------------82
Load more

Recommended publications
  • Upper and Lower Case Letters to Be Used
    Optimizing Foreign Gene Expression in Recombinant Fowl Adenovirus 9 Vectors by James Ackford A Thesis presented to The University of Guelph In partial fulfilment of requirements for the degree of Master of Science in Pathobiology Guelph, Ontario, Canada © James Ackford, August, 2015 ABSTRACT Optimizing Foreign Gene Expression in Recombinant Fowl Adenovirus 9 Vectors James Ackford Advisors: University of Guelph, 2015 Dr. Éva Nagy and Dr. Peter Krell Our laboratory focuses on the molecular characterization of a non-pathogenic strain of fowl adenovirus (FAdV) 9 and its development as a versatile vaccine vector platform. The objectives of this study were to optimize transgene expression by recombinant viruses. High expression promoters and a post-transcriptional regulatory element were evaluated for their ability to improve expression of enhanced green fluorescent protein (EGFP) in recombinant FAdVs. These findings were compared to our current system that employs the human cytomegalovirus (CMV) promoter to express a transgene. EGFP expression was assessed by fluorometry and Western blots. A synthetic CMV enhancer/chicken β-actin (CAG) promoter and the human elongation factor 1 alpha (EF1α) promoter significantly increased expression of EGFP compared to the CMV promoter. However, expression was significantly decreased in the presence of woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). The results provide novel insight into avian vaccine design and optimization of transgene expression by FAdV vectors. ACKNOWLEDGEMENTS First and foremost I would like to thank my co-advisors, Drs. Éva Nagy and Peter Krell, for the continued support and guidance over the years. Their advice, both scientific and personal, has helped me grow as a researcher and professional.
    [Show full text]
  • Les Résultats 2020 En Génomique Des Virus Aviaires
    Centre de Recherche en Infectiologie Porcine et Avicole Swine and Poultry Infectious Diseases research Center Les résultats 2020 de génomique des virus aviaires au Québec Carl A. Gagnon, DMV, PhD, Professeur titulaire, Laboratoire de diagnostic moléculaire (LDM), Réovirus (haut) et poxvirus (bas) aviaires. Image de C. Provost. Service de virologie, de sérologie aviaire et de de microscopie électronique, FMV. UdeM. séquençage à haut débit vétérinaire (LSHDV) Faculté de médecine vétérinaire (FMV), Poste: 8681 Courriel: [email protected] Séquençage du génome entier des pathogènes aviaires http://servicedediagnostic.com/personnel-et- laboratoires/service-de-sequencage/ Tableau 1 – Liste des pathogènes aviaires identifiés au Québec en 2020 par méthode moléculaire au Service de diagnostic Catégorie de Nom Abréviation Maladie pathogènes Bactéries Genre Espèce Chlamydophila spp a C. psittaci Chlamydiose aviairec Escherichia coli E. coli Colibacillose Mycobacterium spp b M. avium Tuberculose aviaire Mycoplasma gallisepticum M. gallisepticum (ou MG) Maladie respiratoire chronique Mycoplasma synoviae M. synoviae (ou MS) Synovite infectieuse Parasite Genre Espèce Histomonas meleagridis H meleagridis Histomonose (peut avoir différents noms selon le syndrome observé) Pathogènes Virus Genre (Espèce) Atadenovirus (Duck atadenovirus A; DAdV-1 Syndrome de la chute de pontec anciennement Duck adenovirus-1) identifiés par PCR Aviadenovirus (Fowl aviadenovirus A to FAdV Hépatite à corps d'inclusion E; anciennement Fowl adenovirus A to E) Avibirnavirus
    [Show full text]
  • 1 Chapter I Overall Issues of Virus and Host Evolution
    CHAPTER I OVERALL ISSUES OF VIRUS AND HOST EVOLUTION tree of life. Yet viruses do have the This book seeks to present the evolution of characteristics of life, can be killed, can become viruses from the perspective of the evolution extinct and adhere to the rules of evolutionary of their host. Since viruses essentially infect biology and Darwinian selection. In addition, all life forms, the book will broadly cover all viruses have enormous impact on the evolution life. Such an organization of the virus of their host. Viruses are ancient life forms, their literature will thus differ considerably from numbers are vast and their role in the fabric of the usual pattern of presenting viruses life is fundamental and unending. They according to either the virus type or the type represent the leading edge of evolution of all of host disease they are associated with. In living entities and they must no longer be left out so doing, it presents the broad patterns of the of the tree of life. evolution of life and evaluates the role of viruses in host evolution as well as the role Definitions. The concept of a virus has old of host in virus evolution. This book also origins, yet our modern understanding or seeks to broadly consider and present the definition of a virus is relatively recent and role of persistent viruses in evolution. directly associated with our unraveling the nature Although we have come to realize that viral of genes and nucleic acids in biological systems. persistence is indeed a common relationship As it will be important to avoid the perpetuation between virus and host, it is usually of some of the vague and sometimes inaccurate considered as a variation of a host infection views of viruses, below we present some pattern and not the basis from which to definitions that apply to modern virology.
    [Show full text]
  • Downloaded from Transcriptome Shotgun Assembly (TSA) Database on 29 November 2020 (Ftp://Ftp.Ddbj.Nig.Ac.Jp/Ddbj Database/Tsa/, Table S3)
    viruses Article Discovery and Characterization of Actively Replicating DNA and Retro-Transcribing Viruses in Lower Vertebrate Hosts Based on RNA Sequencing Xin-Xin Chen, Wei-Chen Wu and Mang Shi * School of Medicine, Sun Yat-sen University, Shenzhen 518107, China; [email protected] (X.-X.C.); [email protected] (W.-C.W.) * Correspondence: [email protected] Abstract: In a previous study, a metatranscriptomics survey of RNA viruses in several important lower vertebrate host groups revealed huge viral diversity, transforming the understanding of the evolution of vertebrate-associated RNA virus groups. However, the diversity of the DNA and retro-transcribing viruses in these host groups was left uncharacterized. Given that RNA sequencing is capable of revealing viruses undergoing active transcription and replication, we collected previously generated datasets associated with lower vertebrate hosts, and searched them for DNA and retro-transcribing viruses. Our results revealed the complete genome, or “core gene sets”, of 18 vertebrate-associated DNA and retro-transcribing viruses in cartilaginous fishes, ray- finned fishes, and amphibians, many of which had high abundance levels, and some of which showed systemic infections in multiple organs, suggesting active transcription or acute infection within the host. Furthermore, these new findings recharacterized the evolutionary history in the families Hepadnaviridae, Papillomaviridae, and Alloherpesviridae, confirming long-term virus–host codivergence relationships for these virus groups.
    [Show full text]
  • AAAP Proceedings
    uAAAP Symposium AUGUST 6-9 2016 Emerging and Reemerging Zoonotic Diseases u AAAP Scientific Program includes 148 oral presentations and 82 poster presentations SymposiumAAAP & Scientific Program THE AMERICAN ASSOCIATION OF AVIAN PATHOLOGISTS promotes scientific knowledge to enhance the health, well-being, and productivity of poultry to provide safe and abundant food for the world. San Antonio, TX August 6-9, 2016 1 of 26 AAAP 2016 Annual Meeting San Antonio Convention Center San Antonio, Texaz, USA August 6-9, 2016 Saturday, August 6, 2016 (7:30AM – 12:30PM) Monday, August 8, 2016 (11:15AM – 11:45AM) AAAP Symposium Lasher-Eckroade History Lecture: The History of the First Emerging and Reemerging Zoonotic Diseases Generation Marek’s Disease Vaccines: The Science and San Antonio Convention Center, Room 221AB Little Known Facts Karel Schat Saturday, August 6, 2016 (2:00PM – 5:00PM) Cornell University, Ithaca, NY AAAP Committee Meetings San Antonio Convention Center, Room 221AB Marriot River Center Hotel Monday, August 8, 2016 (12:00PM – 2:00PM) Saturday, August 6, 2016 (5:00PM – 7:00PM) AAAP Awards Luncheon AAAP New Member Meet and Greet Marriot River Center Hotel, Salon E AAAP Members are invited to welcome new members Marriot River Center Hotel, Salon I Monday, August 8, 2016 (4:30PM – 5:30PM) Wine & Cheese Social in Poster Room Sunday-Tuesday, August 7-9, 2016 (8:00AM – 5:30PM) Poster Presenters will be present at their posters during AAAP Scientific Program this time. San Antonio Convention Center San Antonio Convention Center, Room 220 Sessions A: Room 221AB Sessions B: Room 221CD Tuesday, August 9, 2016 (8:15AM – 8:30AM) Poster Room: 220 Richard B.
    [Show full text]
  • Adenoviruses in Côte D`Ivoire: Investigation of Diversity and Interspecies Transmission
    Adenoviruses in Côte d`Ivoire: investigation of diversity and interspecies transmission von Maude Suzanne Pauly Inaugural-Dissertation zur Erlangung der tiermedizinischen Doktorwürde der Tierärztlichen Fakultät der Ludwig-Maximilians-Universität München Adenoviruses in Côte d`Ivoire: investigation of diversity and interspecies transmission von Maude Suzanne Pauly aus Strasburg (Frankreich) München 2015 Aus dem Veterinärwissenschaftlichen Department der Tierärztlichen Fakultät der Ludwig-Maximilians-Universität München Lehrstuhl für Virologie Institut für Infektionsmedizin und Zoonosen Arbeit angefertigt unter der Leitung von Prof. Dr. Sutter Angefertigt am Robert Koch-Institut Mentoren: Dr. Leendertz und Dr. Ehlers Gedruckt mit Genehmigung der Tierärztlichen Fakultät der Ludwig Maximilians Universität München Dekan: Univ.-Prof. Dr. Joachim Braun Berichterstatter: Prof. Dr.Gerd Sutter Korreferent/en: Prof. Dr. Andrea Fischer Tag der Promotion: 31.Januar 2015 This thesis is dedicated to Mamama and Metto, who will always walk beside me and will stay forewer in my heart. Three first author publications based on data collected and analyses performed within the framework of the thesis, were published: “High prevalence and diversity of species D AdV (HAdV D) in human populations of four Sub-Saharan countries.” (Pauly et al. 2014), “Adenovirus in Rural Côte D`Ivoire: High Diversity and Cross-Species Detection.”(Pauly et al. 2015) and “Contact with non- human primates and risk factors for zoonotic disease emergence in the Taï region, Côte d`Ivoire.”(Mossoun et al. 2015) (shared first authorship). Several sections of the papers were slightly adopted or directly copied in this thesis. A student in biotechnology of the University of Applied Sciences (Beuth Hochschule für Technik, Berlin), Nanina Buchwald, did some of the analyses (first AdV screening of the poultry and of the rodents) for her bachelor thesis and was supervised by Maude Pauly and Dr.
    [Show full text]
  • Whole Volume File
    Turk Vet J TURKISH VETERINARY JOURNAL Journal of the Faculty of Veterinary Medicine, Sivas Cumhuriyet University Aims and Scope Turkish Veterinary Journal (Turk Vet J), aims to publish articles (original research article, short communication, letters to editor, review article, and case report) on Veterinary basic sciences, clinic or preclinical sciences, zootechnics, animal nutrition, food hygiene and technology both in Turkish or English. Turkish Veterinary Journal (Turk Vet J) is an international, double peer reviewing scientific journal that publishes by Sivas Cumhuriyet University, Faculty of Veterinary Medicine. Manuscript will be publish should not published before in elsewhere and should be based on the research (except review articles). Turk Vet J is published three in a year and an open access scientific journal. Special issues may be publishes by the decision of the journal administration. The journal accepts English or Turkish manuscripts. Turk Vet J doesn't asks fee for the processing the article. Amaç ve Kapsam Turkish Veterinary Journal (Turk Vet J), Türkçe veya İngilizce olarak, temel bilimler, klinik ve klinik öncesi bilimler, zootekni, hayvan besleme ile gıda hijyeni ve teknolojisi alanlarında makaleleri (araştırma makalesi, kısa bildiri, editöre mektup, derleme ve vaka takdimi türlerinde) yayımlamayı amaçlar. Turkish Veterinary Journal (Turk Vet J), Sivas Cumhuriyet Üniversitesi, Veteriner Fakültesi tarafından yayımlanan, uluslararası, çift hakemli bilimsel bir dergidir. Dergiye, ulusal ya da uluslararası ilgi ve uygulama içiren ve yeni bilgiler barındıran makaleler sunulabilir. Yayınlanacak makalelerin, daha önceden yayımlanmamış ve araştırma sonuçlarına dayalı olması gerekmektedir (derleme makaleleri hariç). Turk Vet J açık erişim sağlamak üzere yılda üç defa online olarak yayınlanır. Dergi yönetiminin kararları doğrultusunda özel ya da ek sayılar yayımlanabilir.
    [Show full text]
  • Faecal Virome Analysis of Wild Animals from Brazil
    viruses Article Faecal Virome Analysis of Wild Animals from Brazil Matheus A. Duarte 1,2, João M. F. Silva 2 , Clara R. Brito 1, Danilo S. Teixeira 3, Fernando L. Melo 4 , Bergmann M. Ribeiro 2 , Tatsuya Nagata 2 and Fabrício S. Campos 5,* 1 Faculdade de Agronomia e Veterinária, Universidade de Brasília, Brasília-DF 70.910-900, Brazil 2 Departamento de Biologia Celular, Instituto de Biologia, Universidade de Brasília, Brasília-DF 70.910-900, Brazil 3 Núcleo de Atendimento e Pesquisa de Animais Silvestres, Universidade Estadual de Santa Cruz, Ilhéus-BA 45.662-900, Brazil 4 Departamento de Fitopatologia, Instituto de Biologia, Universidade de Brasília, Brasília-DF 70.910-900, Brazil 5 Laboratório de Bioinformática e Biotecnologia, Campus de Gurupi, Universidade Federal do Tocantins, Tocantins-TO 77.410-570, Brazil * Correspondence: [email protected] Received: 30 July 2019; Accepted: 28 August 2019; Published: 30 August 2019 Abstract: The Brazilian Cerrado fauna shows very wide diversity and can be a potential viral reservoir. Therefore, the animal’s susceptibility to some virus can serve as early warning signs of potential human virus diseases. Moreover, the wild animal virome of this biome is unknown. Based on this scenario, high-throughput sequencing contributes a robust tool for the identification of known and unknown virus species in this environment. In the present study, faeces samples from cerrado birds (Psittacara leucophthalmus, Amazona aestiva, and Sicalis flaveola) and mammals (Didelphis albiventris, Sapajus libidinosus, and Galictis cuja) were collected at the Veterinary Hospital, University of Brasília. Viral nucleic acid was extracted, submitted to random amplification, and sequenced by Illumina HiSeq platform.
    [Show full text]
  • Wildlife Virology: Emerging Wildlife Viruses of Veterinary and Zoonotic Importance
    Wildlife Virology: Emerging Wildlife Viruses of Veterinary and Zoonotic Importance Course #: VME 6195/4906 Class periods: MWF 4:05-4:55 p.m. Class location: Veterinary Academic Building (VAB) Room V3-114 and/or Zoom Academic Term: Spring 2021 Instructor: Andrew Allison, Ph.D. Assistant Professor of Veterinary Virology Department of Comparative, Diagnostic, and Population Medicine College of Veterinary Medicine E-mail: [email protected] Office phone: 352-294-4127 Office location: Veterinary Academic Building V2-151 Office hours: Contact instructor through e-mail to set up an appointment Teaching Assistants: NA Course description: The emergence of viruses that cause disease in animals and humans is a constant threat to veterinary and public health and will continue to be for years to come. The vast majority of recently emerging viruses that have led to explosive outbreaks in humans are naturally maintained in wildlife species, such as influenza A virus (ducks and shorebirds), Ebola virus (bats), Zika virus (non-human primates), and severe acute respiratory syndrome (SARS) coronaviruses (bats). Such epidemics can have severe psychosocial impacts due to widespread morbidity and mortality in humans (and/or domestic animals in the case of epizootics), long-term regional and global economic repercussions costing billions of dollars, in addition to having adverse impacts on vulnerable wildlife populations. Wildlife Virology is a 3-credit (3 hours of lecture/week) undergraduate/graduate-level course focusing on pathogenic viruses that are naturally maintained in wildlife species which are transmissible to humans, domestic animals, and other wildlife/zoological species. In this course, we will cover a comprehensive and diverse set of RNA and DNA viruses that naturally infect free-ranging mammals, birds, reptiles, amphibians, and fish.
    [Show full text]
  • Supplemental Material.Pdf
    SUPPLEMENTAL MATERIAL A cloud-compatible bioinformatics pipeline for ultra-rapid pathogen identification from next-generation sequencing of clinical samples Samia N. Naccache1,2, Scot Federman1,2, Narayanan Veerarghavan1,2, Matei Zaharia3, Deanna Lee1,2, Erik Samayoa1,2, Jerome Bouquet1,2, Alexander L. Greninger4, Ka-Cheung Luk5, Barryett Enge6, Debra A. Wadford6, Sharon L. Messenger6, Gillian L. Genrich1, Kristen Pellegrino7, Gilda Grard8, Eric Leroy8, Bradley S. Schneider9, Joseph N. Fair9, Miguel A.Martinez10, Pavel Isa10, John A. Crump11,12,13, Joseph L. DeRisi4, Taylor Sittler1, John Hackett, Jr.5, Steve Miller1,2 and Charles Y. Chiu1,2,14* Affiliations: 1Department of Laboratory Medicine, UCSF, San Francisco, CA, USA 2UCSF-Abbott Viral Diagnostics and Discovery Center, San Francisco, CA, USA 3Department of Computer Science, University of California, Berkeley, CA, USA 4Department of Biochemistry, UCSF, San Francisco, CA, USA 5Abbott Diagnostics, Abbott Park, IL, USA 6Viral and Rickettsial Disease Laboratory, California Department of Public Health, Richmond, CA, USA 7Department of Family and Community Medicine, UCSF, San Francisco, CA, USA 8Viral Emergent Diseases Unit, Centre International de Recherches Médicales de Franceville, Franceville, Gabon 9Metabiota, Inc., San Francisco, CA, USA 10Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México 11Division of Infectious Diseases and International Health and the Duke Global Health
    [Show full text]
  • The Production of Avian Vaccines to Increase Supply Security
    1 Continuous cell lines from the muscovy duck as replacement for primary cells in 2 the production of avian vaccines to increase supply security 3 4 Abstract 5 6 Veterinary vaccines contribute to food security, interrupt zoonotic transmissions, and help to maintain 7 overall health in livestock. Although vaccines are usually cost-effective, their adoption depends on a 8 multitude of factors. Because poultry vaccines are usually given to birds with a short life span, very low 9 production cost per dose is one important challenge. Other hurdles are to ensure a consistent and 10 reliable supply of very large number of doses, and to have flexible production processes to 11 accommodate a range of different pathogens and dosage requirements. Most poultry vaccines are 12 currently being produced on primary avian cells derived from chicken or waterfowl embryos. This 13 production system is associated with high costs, logistic complexities, rigid intervals between harvest and 14 production, and supply limitations. We investigated whether the continuous cell lines Cairina retina and 15 CR.pIX may provide a substrate independent of primary cell cultures or embryonated eggs. Viruses 16 examined for replication in these cell lines are strains associated with, or contained in vaccines against 17 egg drop syndrome, Marek's disease, Newcastle disease, avian influenza, infectious bursal disease and 18 Derzsy's disease. Each of the tested viruses required the development of unique conditions for 19 replication that are described here and can be used to generate material for in vivo efficacy studies and 20 to accelerate transfer of the processes to larger production volumes.
    [Show full text]
  • Genetic Assays for Detecting Viral Recombination Rate
    (19) *EP003024948B1* (11) EP 3 024 948 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: C12Q 1/6827 (2018.01) C12Q 1/70 (2006.01) 15.01.2020 Bulletin 2020/03 (86) International application number: (21) Application number: 14829649.4 PCT/US2014/048301 (22) Date of filing: 25.07.2014 (87) International publication number: WO 2015/013681 (29.01.2015 Gazette 2015/04) (54) GENETIC ASSAYS FOR DETECTING VIRAL RECOMBINATION RATE GENETISCHE TESTS ZUR BESTIMMUNG EINER VIRALEN REKOMBINATIONSFREQUENZ DOSAGES GÉNÉTIQUES POUR DÉTERMINER UNE FREQUENCE DE LA RECOMBINATION VIRALE (84) Designated Contracting States: • A. D. TADMOR ET AL: "Probing Individual AL AT BE BG CH CY CZ DE DK EE ES FI FR GB Environmental Bacteria for Viruses by Using GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO Microfluidic Digital PCR", SCIENCE, vol. 333, no. PL PT RO RS SE SI SK SM TR 6038, 1 July 2011 (2011-07-01), pages 58-62, XP055344735, ISSN: 0036-8075, DOI: (30) Priority: 25.07.2013 US 201361858311 P 10.1126/science.1200758 -& A. D. TADMOR ET 01.11.2013 US 201361899027 P AL: "Probing Individual Environmental Bacteria for Viruses by Using Microfluidic Digital PCR - (43) Date of publication of application: Supporting Online Material", SCIENCE, vol. 333, 01.06.2016 Bulletin 2016/22 no. 6038, 30 June 2011 (2011-06-30), pages 1-48, XP055344921, ISSN: 0036-8075, DOI: (73) Proprietor: Bio-rad Laboratories, Inc. 10.1126/science.1200758 Hercules, CA 94547 (US) • K.
    [Show full text]