DOCSLIB.ORG

Francisella Novicida Mice Infected with Deficient

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Francisella Novicida Mice Infected with Deficient ASC Controls IFN-γ Levels in an IL-18− Dependent Manner in Caspase-1−Deficient Mice Infected with Francisella novicida This information is current as Roberto Pierini, Magali Perret, Sophia Djebali, Carole Juruj, of September 27, 2021. Marie-Cécile Michallet, Irmgard Förster, Jacqueline Marvel, Thierry Walzer and Thomas Henry J Immunol 2013; 191:3847-3857; Prepublished online 23 August 2013; doi: 10.4049/jimmunol.1203326 Downloaded from http://www.jimmunol.org/content/191/7/3847 Supplementary http://www.jimmunol.org/content/suppl/2013/08/23/jimmunol.120332 Material 6.DC1 http://www.jimmunol.org/ References This article cites 60 articles, 21 of which you can access for free at: http://www.jimmunol.org/content/191/7/3847.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on September 27, 2021 • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2013 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology ASC Controls IFN-g Levels in an IL-18–Dependent Manner in Caspase-1–Deficient Mice Infected with Francisella novicida Roberto Pierini,*,†,‡,x Magali Perret,*,†,‡,x Sophia Djebali,*,†,‡,x Carole Juruj,*,†,‡,x Marie-Ce´cile Michallet,*,†,‡,x Irmgard Fo¨rster,{,1 Jacqueline Marvel,*,†,‡,x Thierry Walzer,*,†,‡,x and Thomas Henry*,†,‡,x The inflammasome is a signaling platform that is central to the innate immune responses to bacterial infections. Francisella tularensis is a bacterium replicating within the host cytosol. During F. tularensis subspecies novicida infection, AIM2, an inflam- masome receptor sensing cytosolic DNA, activates caspase-1 in an ASC-dependent manner, leading to both pyroptosis and release of the proinflammatory cytokines IL-1b and IL-18. Activation of this canonical inflammasome pathway is key to limit F. novicida Downloaded from infection. In this study, by comparing the immune responses of AIM2 knockout (KO), ASCKO, and Casp1KO mice in response to F. novicida infection, we observed that IFN-g levels in the serum of Casp1KO mice were much higher than the levels observed in AIM2KO and ASCKO mice. This difference in IFN-g production was due to a large production of IFN-g by NK cells in Casp1KO mice that was not observed in ASCKO mice. The deficit in IFN-g production observed in ASCKO mice was not due to a reduced Dock2 expression or to an intrinsic defect of ASCKO NK cells. We demonstrate that in infected Casp1KO mice, IFN-g production is due to an ASC-dependent caspase-1–independent pathway generating IL-18. Furthermore, we present in vitro data suggesting http://www.jimmunol.org/ that the recently described AIM2/ASC/caspase-8 noncanonical pathway is responsible for the caspase-1–independent IL-18 releasing activity. To our knowledge, this study is the first in vivo evidence of an alternative pathway able to generate in a caspase-1–independent pathway bioactive IL-18 to boost the production of IFN-g, a cytokine critical for the host antibacterial response. The Journal of Immunology, 2013, 191: 3847–3857. he inflammasome (1) is an innate immune platform ac- masome receptor detecting DNA within the host cytosol (2–5). We tivated in response to danger signals and infections and and others have recently described that the AIM2 inflammasome T leading to caspase-1 activation. Caspase-1 is an inflam- is activated following infections with “cytosolic” bacteria such as by guest on September 27, 2021 matory caspase leading to the processing of pro–IL-1b and pro–IL- Listeria monocytogenes and Francisella tularensis (6–9). 18 and the release of the corresponding cytokines. Furthermore, F. tularensis is a highly infectious bacterium that causes tula- caspase-1 also triggers cell death in a process termed pyroptosis. remia in humans (10). Francisella novicida (also known as F. Caspase-1 activation takes place within a multiprotein complex, the tularensis subspecies novicida) is considered nonpathogenic for inflammasome, which includes a receptor, and an adaptor, ASC. humans and is used as a model to study highly virulent F. tular- Several inflammasomes have been described depending on the ensis infections. In mice, F. novicida is found mostly in myeloid detected ligand and on the receptor involved. AIM2 is an inflam- cells such as macrophages and neutrophils (11). Its ability to cause disease is tightly linked to its ability to rapidly escape from the phagosome into the host cytosol where it can replicate to very high *Centre International de Recherche en Infectiologie, Universite´ de Lyon, Lyon numbers (12, 13). 69007, France; †INSERM, Unite´ 851, Lyon 69007, France; ‡Centre National de la Recherche Scientifique, Unite´ Mixte de Recherche 5308, Lyon 69007, France; xEcole Infection with wild-type (WT) F. novicida leads to the activation Normale Supe´rieure de Lyon, Lyon 69342, France; and {Department of Molecular of the AIM2 inflammasome (7–9). Mice deficient for AIM2, ASC, Immunology, Leibniz Research Institute for Environmental Medicine, 40225 Du¨ssel- or for both caspase-1 and caspase-11 (14) are very susceptible to F. dorf, Germany novicida infection. This high susceptibility of ASC knockout (KO) 1Current address: Immunology and Environment, Life and Medical Sciences Insti- tute, University of Bonn, Bonn, Germany. and Casp1/11 double KO mice is due to both caspase-1–mediated Received for publication December 4, 2012. Accepted for publication July 24, 2013. control of IL-1b and IL-18 and to caspase-1–mediated cell death (15–17). In contrast to other intracellular bacteria, detection of This work was supported by Marie Curie International Reintegration Grant PIRG07- GA-2010-268399 and by a Fondation Innovations en Infectiologie Young Inves- F. novicida by the inflammasome in murine cells seems to be ex- tigator grant. This work was performed within the framework of the Laboratoire clusively dependent on AIM2 (8). F. novicida infection is thus d’Excellence eco-evolutionary dynamics of infectious diseases (ECOFECT Grant ANR- 11-LABX-0042) of the Universite´ de Lyon, within the program “Investissements d’Avenir” emerging as a very good model to study both in vitro and in vivo this (Grant ANR-11-IDEX-0007) operated by the French National Research Agency. innate immune response (8, 9, 18). Caspase-11 is dispensable for Address correspondence and reprint requests to Dr. Thomas Henry, INSERM, Unite´ caspase-1 activation in response to F. novicida infection (14, 19). 851, 21 Avenue Tony Garnier, Lyon 69007, France. E-mail address: thomas.henry@ The canonical inflammasome pathway leads to caspase-1 acti- inserm.fr vation. However, we and others have shown that the inflammasome The online version of this article contains supplemental material. adaptor ASC could have caspase-1–independent activities. In vitro, Abbreviations used in this article: BMM, bone marrow macrophage; KO, knockout; ASC can recruit and activate apoptotic caspases leading to mac- PI, postinfection; WT, wild-type. rophage apoptosis (18, 20–23). In several noninfectious inflam- Copyright Ó 2013 by The American Association of Immunologists, Inc. 0022-1767/13/$16.00 matory models, ASC contributes to inflammation in a caspase-1– www.jimmunol.org/cgi/doi/10.4049/jimmunol.1203326 3848 ASC/IL-18–DEPENDENT REGULATION OF IFN-g IN Casp1KO MICE dependent and caspase-1–independent manner (24–26). Similarly, in FACS buffer and Live/Dead fixable dead cell stain kit (Invitrogen) in during chronic Mycobacterium tuberculosis infection in mice, PBS containing 2.5 mM EDTA. Peripheral blood samples were collected ASC participates in granuloma formation mechanisms indepen- in EDTA-containing tubes and incubated for 30 min at 4˚C with mAbs cocktails. dently of caspase-1 (27). Recently, Dock2 expression was shown Phenotyping and analysis of NK cell maturation and function was to be deficient in several but not all ASC-deficient mouse lines, performed as previously described (32, 33). When applicable, spleen leading to a severe immune deficit in ASCKO mouse lines ex- single-cell suspensions were incubated for 4 h with IL-12 (20 ng/ml), IL- pressing low levels of Dock2 protein (28, 29). It is still unclear 12 plus IL-2 (3000 U/ml; PeproTech), IL-12 plus IL-18 (20 mg/ml; MBL International), or on coated Abs anti-Ly49 (4E5) or anti-NKp46 (29A1.4) whether some of the phenotypes described above relate to a deficit in the presence of monensin and anti-CD107a (Lamp-1) Ab (BD Bio- in Dock2. sciences). Cells were analyzed using a FACSCanto II flow cytometer In this study, using F. novicida infection as a model to engage (Becton Dickinson, San Jose, CA). Data were processed using FlowJo the AIM2/ASC inflammasome components, we compared the im- software (Tree Star, Asland, OR). mune responses of WT mice with the ones of mice deficient for RNA and quantitative real-time RT-PCR ASC (ASCKO) or for both caspase-1 and caspase-11 (termed Casp1KO). We found that ASC controls IFN-g secretion by NK Approximately a third of the spleen was ground in 1 ml TRIzol. RNA was extracted using chloroform followed by 70% ethanol-mediated precipitation. cells in a caspase-1–dependent and caspase-1–independent man- RNA was then isolated using the RNeasy Mini kit (Qiagen). Reverse tran- ner. This was independent of Dock2 because our ASCKO mouse scription and quantitative PCR were performed using Improm (Promega) line had normal Dock2 protein levels.
Load more

Recommended publications
  • Francisella Spp. Infections in Farmed and Wild Fish. ICES CM 2008/D:07
    ICES CM 2008/D:07 Francisella spp. infections in farmed and wild fish Duncan J. Colquhoun1, Adam Zerihun2 and Jarle Mikalsen3 National Veterinary Institute, Section for Fish Health, Ullevaalsveien 68, 0454 Oslo, Norway 1 tel: +47 23 21 61 41; fax: +47 23 21 61 01; e-mail: [email protected] 2 tel: +47 23 21 61 08; fax: +47 23 21 61 01; e-mail: [email protected] 3 tel: +47 23 21 61 55; fax: +47 23 21 61 01; e-mail: [email protected] Abstract Bacteria within the genus Francisella are non-motile, Gram-negative, strictly aerobic, facultatively intracellular cocco-bacilli. While the genus includes pathogens of warm-blooded animals including humans, and potential bioterror agents, there is also increasing evidence of a number of as yet unrecognised environmental species. Due to their nutritionally fastidious nature, bacteria of the genus Francisella are generally difficult to culture, and growth is also commonly inhibited by the presence of other bacteria within sample material. For these reasons, Francisella-related fish disease may be under-diagnosed. Following the discovery in 2004/2005 that a granulomatous disease in farmed and wild Atlantic cod (Gadus morhua) is caused by a previously undescribed member of this genus (Francisella philomiragia subsp. noatunensis), similar diseases have been identified in fish in at least seven countries around the world. These infections affect both freshwater and marine fish species and involve bacteria more or less closely related to F. philomiragia subsp. philomiragia, an opportunistic human pathogen. Recent work relating to characterisation of the disease/s, classification of fish pathogenic Francisella spp.
    [Show full text]
  • Bioinformatics Resource Centers Systems Biology (Brcs) Centers
    Fondation Merieux – J Craig Venter Institute Bioinformatics Workshop December 5 – 8, 2017 Module 3: Genomic Data & Sequence Annotations in Public Databases NIH/NIAID Genomics and Bioinformatics Program SlideSource:A.S.Fauci SlideSource:A.S.Fauci Conducts and supports basic and applied research to better understand, treat, and ultimately prevent infectious, immunologic, and allergic diseases. NIAIDGenomicsProgram Proteomics Systems Sequencing Functional Structural Biology Genomics Genomics Genomic Clinical Functional Systems Sequencing Proteomics Structural Genomic Biology Centers Centers Genomics Research Centers Centers Centers Bioinformatics BioinformaticsResource Centers GenomicResearchResources Genomic/OmicsDataSets,Databases,BioinformaticsTools,Biomarkers,3DStructures,ProteinClones,PredictiveModels Toaddresskeyquestionsin microbiologyandinfectious disease NIAID Genome Sequencing Center Influenza Genome Sequencing Project at JCVI • 2004: 80 influenza genomes in GenBank • 3OCT2017: ~20,000 influenza genomes sequenced at JCVI • 75% complete influenza genomes in GenBank by JCVI Slide source: Maria Giovanni * Genome Sequencing Centers Bioinformatics Resource Centers Systems Biology (BRCs) Centers Structure Genomics Centers Clinical Proteomics Centers Courtesy of Alison Yao, DMID *Bioinformatics Resource Centers (BRCs) Goal: Provide integrated bioinformatics resources in support of basic and applied infectious diseases research • Data and metadata management and integration solutions • Computational analysis and visualization tools • Work
    [Show full text]
  • Francisella Tularensis Subspecies Holarctica and Tularemia in Germany
    microorganisms Review Francisella tularensis Subspecies holarctica and Tularemia in Germany 1, 2, 3 1 1 Sandra Appelt y, Mirko Faber y , Kristin Köppen , Daniela Jacob , Roland Grunow and Klaus Heuner 3,* 1 Centre for Biological Threats and Special Pathogens (ZBS 2), Robert Koch Institute, 13353 Berlin, Germany; [email protected] (S.A.); [email protected] (D.J.); [email protected] (R.G.) 2 Gastrointestinal Infections, Zoonoses and Tropical Infections (Division 35), Department for Infectious Disease Epidemiology, Robert Koch Institute, 13353 Berlin, Germany; [email protected] 3 Cellular Interactions of Bacterial Pathogens, ZBS 2, Robert Koch Institute, 13353 Berlin, Germany; [email protected] * Correspondence: [email protected]; Tel.: +49-301-8754-2226 These authors contributed equally to this work. y Received: 27 August 2020; Accepted: 18 September 2020; Published: 22 September 2020 Abstract: Tularemia is a zoonotic disease caused by Francisella tularensis a small, pleomorphic, facultative intracellular bacterium. In Europe, infections in animals and humans are caused mainly by Francisella tularensis subspecies holarctica. Humans can be exposed to the pathogen directly and indirectly through contact with sick animals, carcasses, mosquitoes and ticks, environmental sources such as contaminated water or soil, and food. So far, F. tularensis subsp. holarctica is the only Francisella species known to cause tularemia in Germany. On the basis of surveillance data, outbreak investigations, and literature, we review herein the epidemiological situation—noteworthy clinical cases next to genetic diversity of F. tularensis subsp. holarctica strains isolated from patients. In the last 15 years, the yearly number of notified cases of tularemia has increased steadily in Germany, suggesting that the disease is re-emerging.
    [Show full text]
  • Isolation of Francisella Tularensis from Skin Ulcer After a Tick Bite, Austria, 2020
    microorganisms Case Report Isolation of Francisella tularensis from Skin Ulcer after a Tick Bite, Austria, 2020 Mateusz Markowicz 1,*, Anna-Margarita Schötta 1 , Freya Penatzer 2, Christoph Matscheko 2, Gerold Stanek 1, Hannes Stockinger 1 and Josef Riedler 2 1 Center for Pathophysiology, Infectiology and Immunology, Institute for Hygiene and Applied Immunology, Medical University of Vienna, Kinderspitalgasse 15, A-1090 Vienna, Austria; [email protected] (A.-M.S.); [email protected] (G.S.); [email protected] (H.S.) 2 Kardinal Schwarzenberg Klinikum, Kardinal Schwarzenbergplatz 1, A-5620 Schwarzach, Austria; [email protected] (F.P.); [email protected] (C.M.); [email protected] (J.R.) * Correspondence: [email protected]; Tel.: +43-1-40160-33023 Abstract: Ulceroglandular tularemia is caused by the transmission of Francisella tularensis by arthro- pods to a human host. We report a case of tick-borne tularemia in Austria which was followed by an abscess formation in a lymph node, making drainage necessary. F. tularensis subsp. holarctica was identified by PCR and multilocus sequence typing. Keywords: tularemia; Francisella tularensis; tick; multi locus sequence typing Depending on the transmission route of Francisella tularensis, tularemia can present Citation: Markowicz, M.; Schötta, as a local infection or a systemic disease [1]. Transmission of the pathogen takes place A.-M.; Penatzer, F.; Matscheko, C.; by contact with infected animals, by bites of arthropods or through contaminated water Stanek, G.; Stockinger, H.; Riedler, J. and soil. Hares and wild rabbits are the main reservoirs of the pathogen in Austria [2].
    [Show full text]
  • Francisella Tularensis
    The Genetic Composition and Diversity of Francisella tularensis Pär Larsson Akademisk avhandling som med vederbörligt tillstånd av rektorsämbetet vid Umeå Universitet för avläggande av medicine doktorsexamen i klinisk mikrobiologi med inriktning mot bakteriologi vid Medicinska fakulteten, framlägges till offentligt försvar vid Institutionen för Klinisk Mikrobiologi, sal E04 byggnad 6, torsdagen den 31 maj 2007, klockan 09.00. Avhandlingen kommer att försvaras på engelska. Fakultetsopponent: Dr. Andrew K Benson Department of Food Science & Technology University of Nebraska–Lincoln Lincoln, Nebraska USA Department of Clinical Microbiology, Clinical Bacteriology Umeå University Umeå 2007 Organization Document type UMEÅ UNIVERSITY DOCTORAL DISSERTATION Department of Clinical Microbiology Date of publication SE-901 87 Umeå, Sweden May 2007 Author Pär Larsson Title The Genetic Composition and Diversity of Francisella tularensis Abstract Francisella tularensis is the causative agent of the debilitating, sometimes fatal zoonotic disease tularemia. Despite all F. tularensis bacteria having very similar genotypes and phenotypes, the disease varies significantly in severity depending on the subspecies of the infectious strain. To date, little information has been available on the genetic makeup of this pathogen, its evolution, and the genetic differences which characterize subspecific lineages. These are the main areas addressed in this thesis. Using the F. tularensis subsp. tularensis SCHU S4 strain as a genetic reference, microarray-based comparative genomic hybridisations were used to investigate the differences in genomic composition of F. tularensis isolates. Overall, the strains analysed were very similar, matching the high degree of conservation previously observed at the sequence level. One striking finding was that subsp. mediasiatica was most similar to subsp. tularensis, despite their natural confinement to Central Asia and North America, respectively.
    [Show full text]
  • Tularemia – Epidemiology
    This first edition of theWHO guidelines on tularaemia is the WHO GUIDELINES ON TULARAEMIA result of an international collaboration, initiated at a WHO meeting WHO GUIDELINES ON in Bath, UK in 2003. The target audience includes clinicians, laboratory personnel, public health workers, veterinarians, and any other person with an interest in zoonoses. Tularaemia Tularaemia is a bacterial zoonotic disease of the northern hemisphere. The bacterium (Francisella tularensis) is highly virulent for humans and a range of animals such as rodents, hares and rabbits. Humans can infect themselves by direct contact with infected animals, by arthropod bites, by ingestion of contaminated water or food, or by inhalation of infective aerosols. There is no human-to-human transmission. In addition to its natural occurrence, F. tularensis evokes great concern as a potential bioterrorism agent. F. tularensis subspecies tularensis is one of the most infectious pathogens known in human medicine. In order to avoid laboratory-associated infection, safety measures are needed and consequently, clinical laboratories do not generally accept specimens for culture. However, since clinical management of cases depends on early recognition, there is an urgent need for diagnostic services. The book provides background information on the disease, describes the current best practices for its diagnosis and treatment in humans, suggests measures to be taken in case of epidemics and provides guidance on how to handle F. tularensis in the laboratory. ISBN 978 92 4 154737 6 WHO EPIDEMIC AND PANDEMIC ALERT AND RESPONSE WHO Guidelines on Tularaemia EPIDEMIC AND PANDEMIC ALERT AND RESPONSE WHO Library Cataloguing-in-Publication Data WHO Guidelines on Tularaemia.
    [Show full text]
  • Communicating in a Crisis: Biological Attack
    2. Use common sense, practice good hygiene and cleanliness to avoid spreading germs. “Communication before, during People who are potentially exposed should: and after a biological attack will 1. Follow instructions of health care providers and other public health officials. NEWS &TERRORISM 2. Expect to receive medical evaluation and treatment. Be prepared for long lines. If COMMUNICATING IN A CRISIS be a critical element in effectively the disease is contagious, persons exposed may be quarantined. A fact sheet from the National Academies and the U.S. Department of Homeland Security responding to the crisis and help­ If people become aware of a suspicious substance nearby, they should: ing people to protect themselves 1. Quickly get away. and recover.” 2. Cover their mouths and noses with layers of fabric that can filter the air but still allow breathing. —A Journalist’s Guide to Covering 3. Wash with soap and water. Bioterrorism (Radio and Television News 4. Contact authorities. BIOLOGICAL ATTACK Director’s Foundation, 2004) 5. Watch TV, listen to the radio, or check the Internet for official news and informa- HUMAN PATHOGENS, BIOTOXINS, tion including the signs and symptoms of the disease, if medications or vaccinations AND AGRICULTURAL THREATS are being distributed, and where to seek medical attention if they become sick. 6. Seek emergency medical attention if they become sick. Table 1. Diseases/Agents Listed by the CDC as Potential WHAT IS IT? Bioterror Threats (as of March 2005). The U.S. Department of Medical Treatment Agriculture maintains lists of animal and plant agents of concern. Table 2 lists general medical treatments for several biothreat agents.
    [Show full text]
  • A Dissertation Entitled Characterization of a Novel
    A Dissertation entitled Characterization of a novel Francisella tularensis Virulence Factor Involved in Cell Wall Repair by Briana Collette Zellner Submitted to the Graduate Faculty as partial fulfillment of the requirements for the Doctor of Philosophy Degree in Biomedical Sciences ___________________________________________ Jason Huntley, Ph.D., Major Advisor ___________________________________________ R. Mark Wooten, Ph.D., Committee Member ___________________________________________ Jyl Matson, Ph.D., Committee Member ___________________________________________ Robert Blumenthal, Ph.D. Committee Member ___________________________________________ R. Travis Taylor, Ph.D., Committee Member ___________________________________________ Cyndee Gruden, PhD, Dean College of Graduate Studies The University of Toledo December 2019 © 2019 Briana Collette Zellner This document is copyrighted material. Under copyright law, no parts of this document may be reproduced without the expressed permission of the author. An Abstract of Characterization of a Novel Francisella tularensis Virulence Factor Involved in Cell Wall Repair by Briana Collette Zellner Submitted to the Graduate Faculty as partial fulfillment of the requirements for the Doctor of Philosophy Degree in Biomedical Sciences The University of Toledo December 2019 Francisella tularensis, the causative agent of tularemia, is one of the most dangerous bacterial pathogens known. F. tularensis has a low infectious dose, is easily aerosolized, and induces high morbidity and mortality; thus, it
    [Show full text]
  • In Vivo and in Vitro Pathogenesis of Francisella Asiatica in Tilapia
    Louisiana State University LSU Digital Commons LSU Doctoral Dissertations Graduate School 2010 In vivo and in vitro pathogenesis of Francisella asiatica in tilapia nilotica (Oreochromis niloticus) Esteban Soto Louisiana State University and Agricultural and Mechanical College, [email protected] Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_dissertations Part of the Veterinary Pathology and Pathobiology Commons Recommended Citation Soto, Esteban, "In vivo and in vitro pathogenesis of Francisella asiatica in tilapia nilotica (Oreochromis niloticus)" (2010). LSU Doctoral Dissertations. 2796. https://digitalcommons.lsu.edu/gradschool_dissertations/2796 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Doctoral Dissertations by an authorized graduate school editor of LSU Digital Commons. For more information, please [email protected]. IN VIVO AND IN VITRO PATHOGENESIS OF FRANCISELLA ASIATICA IN TILAPIA NILOTICA (OREOCHROMIS NILOTICUS) A Dissertation Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Doctor of Philosophy in The Interdepartmental Program in Veterinary Medical Sciences through the Department of Pathobiological Sciences by Esteban Soto Med.Vet., Universidad Nacional-Costa Rica, 2005 M.Sc., Mississippi State University, 2007 August, 2010 ACKNOWLEDGEMENTS The main reason why I’m being able to present this dissertation is because of all the help and advices received by many people along these years. Firstly and foremost I thank my wife Tati for always believing in me and giving me all the support I needed. To my dad and mom, thanks for being a perfect example of integrity and perseverance.
    [Show full text]
  • Francisella Novicida–Causing Two Samples of Blood Cultures from Peripheral Lines Bacteremia in a Woman from Thailand Who Was Receiving Chemotherapy for Ovarian Cancer
    she was treated with lamivudine. A follow-up visit in early Emergence of September showed that her liver function biochemistry re- sults had returned to within normal limits. Chemotherapy Francisella with carboplastin and paclitaxel was then initiated. At the time of admission, 25 days after the start of novicida chemotherapy, the patient had fever (39oC), blood pressure 90/60 mm Hg, and pulse rate 75 beats/min. She also had Bacteremia, an episode of gastrointestinal hemorrhage with melena. It Thailand was believed that fever and gastrointestinal bleeding were complications from chemotherapy; thus, microbiologic Amornrut Leelaporn, Samaporn Yongyod, investigation was not promptly initiated. Abnormal labo- Sunee Limsrivanichakorn, Thitiya Yungyuen, ratory fi ndings included anemia (hemoglobin 80 g/L) and and Pattarachai Kiratisin leukocytosis with marked neutrophilia (Figure). Urine and stool cultures showed insignifi cant growth. We report isolation of Francisella novicida–causing Two samples of blood cultures from peripheral lines bacteremia in a woman from Thailand who was receiving chemotherapy for ovarian cancer. The organism was iso- were obtained using BacT/Alert FA bottles (bioMérieux, lated from blood cultures and identifi ed by 16S rDNA and Durham, NC, USA) on day 10 of hospital admission and PPIase gene analyses. Diagnosis and treatment were de- incubated in the continuous monitoring BacT/Alert 3D sys- layed due to unawareness of the disease in this region. tem (bioMérieux). Both blood culture bottles grew small pleomorphic gram-negative coccobacillus after incubation for 2 days. Samples from positive bottles were subcultured rancisella novicida, a rare human pathogen, has recent- onto 5% (vol/vol) sheep blood agar, MacConkey agar, and Fly been considered to be a subspecies of F.
    [Show full text]
  • Tularemia (Francisella Tularensis)
    SENTINEL LEVEL CLINICAL LABORATORY GUIDELINES FOR SUSPECTED AGENTS OF BIOTERRORISM AND EMERGING INFECTIOUS DISEASES Francisella tularensis American Society for Microbiology (ASM) Revised March 2016 For latest revision, see web site below: https://www.asm.org/Articles/Policy/Laboratory-Response-Network-LRN-Sentinel-Level-C ASM Subject Matter Experts: David Craft, Ph.D. Major Todd Kijek, Ph.D. Penn State Milton S. Hershey US Army Medical Research Institute for Medical Center Infectious Diseases Hershey, PA Ft. Detrick, MD [email protected] [email protected] ASM Sentinel Level Laboratory Protocol Working Group APHL Advisory Committee Vickie Baselski, Ph.D. Barbara Robinson-Dunn, Ph.D. Patricia Blevins, MPH University of Department of Clinical San Antonio Metro Health Tennessee at Pathology District Laboratory Memphis Beaumont Health System [email protected] Memphis, TN Royal Oak, MI [email protected] BRobinson- Erin Bowles [email protected] Wisconsin State Laboratory of David Craft, Ph.D. Hygiene Penn State Milton S. Michael A. Saubolle, Ph.D. [email protected] Hershey Medical Center Banner Health System Hershey, PA Phoenix, AZ Christopher Chadwick, MS [email protected] Mike.Saubolle@bannerhealt Association of Public Health h.com Laboratories Peter H. Gilligan, Ph.D. [email protected] University of North Susan L. Shiflett Carolina Hospitals/ Michigan Department Mary DeMartino, Clinical Microbiology and of Community Health BS, MT(ASCP)SM Immunology Labs Lansing, MI State Hygienic Laboratory at Chapel Hill, NC [email protected] the University of Iowa [email protected] [email protected] Alice Weissfeld, Ph.D. Larry Gray, Ph.D. Microbiology Specialists Inc.
    [Show full text]
  • The Francisella Pathogenicity Island Its Role in Type VI Secretion and Intracellular Infection
    The Francisella Pathogenicity Island Its role in Type VI Secretion and intracellular infection Lena Meyer Department of Clinical Microbiology Umeå 2015 Responsible publisher under swedish law: the Dean of the Medical Faculty This work is protected by the Swedish Copyright Legislation (Act 1960:729) ISBN: 978-91-7601-246-8 ISSN: 0346-6612 Front cover by Lena Meyer. Injection capillary. Elektronisk version tillgänglig på http://umu.diva-portal.org/ Tryck/Printed by: Print & Media Umeå, Sweden 2015 Somewhere, something incredible is waiting to be known. Carl Sagan Für Familie und Freunde. TABLE OF CONTENTS TABLE OF CONTENTS i ABSTRACT iii SAMMANFATTNING PÅ SVENSKA v ABBREVIATIONS vii LIST OF PAPERS ix PAPERS INCLUDED IN THE THESIS ix PAPERS NOT INCLUDED IN THE THESIS ix 1. INTRODUCTION 1 1.1. FRANCISELLA TULARENSIS – AN OVERVIEW 1 1.2. THE INTRACELLULAR LIFESTYLE OF FRANCISELLA TULARENSIS 2 PHAGOSOMAL ESCAPE AND INTRACELLULAR REPLICATION OF FRANCISELLA 3 METABOLIC ADAPTATION 5 INNATE IMMUNE RECOGNITION OF FRANCISELLA 7 1.3. THE FRANCISELLA PATHOGENICITY ISLAND (FPI) 10 1.4 TYPE VI SECRETION SYSTEM (T6SS) 13 CORE COMPONENTS, STRUCTURE AND EFFECTORS 14 A CURRENT MODEL FOR T6S AND ITS REGULATION 17 THE FRANCISELLA T6SS – OUTSIDE THE BOX? 20 2. AIMS OF THE THESIS 22 3. METHODOLOGICAL CONSIDERATIONS 23 MUTAGENESIS AND COMPLEMENTATION 23 CELL INFECTION, INFECTION MODELS AND RESPONSE 24 FRACTIONATION AND BACTERIAL CELL MEMBRANE INTEGRITY 26 PROTEIN-PROTEIN INTERACTION METHODS 26 PROTEIN SECRETION METHODS 28 MICROINJECTION 29 4. RESULTS AND DISCUSSION 31 4.1 CHARACTERIZATION OF FPI MUTANTS (PAPERS I, II AND III) 32 THE SUBCELLULAR LOCALIZATION OF FPI PROTEINS AND THEIR ENGAGEMENT IN PROTEIN-PROTEIN INTERACTIONS 32 NOVEL PHENOTYPES AND NULL MUTANTS – THE IMPORTANCE OF FPI PROTEINS FOR THE INTRACELLULAR GROWTH CYCLE OF F.
    [Show full text]