DOCSLIB.ORG

Sequential Activation of Two Pathogen-Sensing Pathways Required for Type I Interferon Expression and Resistance to an Acute DNA Virus Infection

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Sequential Activation of Two Pathogen-Sensing Pathways Required for Type I Interferon Expression and Resistance to an Acute DNA Virus Infection Article Sequential Activation of Two Pathogen-Sensing Pathways Required for Type I Interferon Expression and Resistance to an Acute DNA Virus Infection Highlights Authors d TLR9, MyD88, STING, and IRF7 are required for IFN-I Ren-Huan Xu, Eric B. Wong, Daniel expression in the dLN Rubio, ..., Shinu John, Mark Shlomchik, Luis J. Sigal d Inflammatory monocytes are the major producers of IFN-I in the dLN Correspondence + [email protected] d TLR9, MyD88, and IRF7 are required in CD11c cells for iMo recruitment to dLNs In Brief d STING-IRF7 are required in iMos for IFN-a and STING-NF-kB How different pathogen-sensing for IFN-b expression in iMos mechanisms contribute to the expression of IFN-I in vivo is unclear. Sigal and colleagues report that in lymph nodes of ectromelia-virus-infected mice, CD11c+ cells use TLR9-MyD88-IRF7 to recruit inflammatory monocytes (iMos). In turn, infected iMos express IFN-a and IFN-b through STING-IRF7 and STING-NF-kB, respectively. Xu et al., 2015, Immunity 43, 1148–1159 December 15, 2015 ª2015 Elsevier Inc. http://dx.doi.org/10.1016/j.immuni.2015.11.015 Immunity Article Sequential Activation of Two Pathogen-Sensing Pathways Required for Type I Interferon Expression and Resistance to an Acute DNA Virus Infection Ren-Huan Xu,1,5 Eric B. Wong,1,6 Daniel Rubio,1,7 Felicia Roscoe,1 Xueying Ma,1 Savita Nair,1 Sanda Remakus,1 Reto Schwendener,2 Shinu John,3 Mark Shlomchik,4 and Luis J. Sigal1,6,* 1Immune Cell Development and Host Defense Program, The Research Institute at Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, PA 19111, USA 2Institute of Molecular Cancer Research, University Zurich, 8057 Zurich, Switzerland 3Department of Laboratory Medicine, Yale University, New Haven, CT 06520, USA 4Department of Immunology, University of Pittsburgh School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15261, USA 5Present address: Novavax Inc., 20 Firstfield Road, Gaithersburg, MD 20878, USA 6Present address: Department of Microbiology and Immunology, Thomas Jefferson University, BLSB 730, 233 South 10th Street, Philadelphia, PA 19107, USA 7Present address: GlaxoSmithKline, 709 Swedeland Road, King of Prussia, PA 19406, USA *Correspondence: [email protected] http://dx.doi.org/10.1016/j.immuni.2015.11.015 SUMMARY and activate innate and adaptive immune cells (Wu and Chen, 2014). Toll-like receptor 9 (TLR9), its adaptor MyD88, the PAMPs in the cell microenvironment are recognized by downstream transcription factor interferon regulato- transmembrane PRRs that reside in the plasma and endosomal ry factor 7 (IRF7), and type I interferons (IFN-I) are all membranes, such as Toll-like receptors (TLRs)—for example, required for resistance to infection with ectromelia vi- TLR9 recognizes double-stranded DNA in endosomes (Ah- rus (ECTV). However, it is not known how or in which mad-Nejad et al., 2002; Hemmi et al., 2000). PAMPs in the cells these effectors function to promote survival. cytosol, on the other hand, are sensed by cytosolic PRRs—for instance, RIG-I-like receptors (RLRs) detect cytosolic viral RNA Here, we showed that after infection with ECTV, species (Goubau et al., 2013; Vabret and Blander, 2013), the TLR9-MyD88-IRF7 pathway was necessary in + whereas DNA-dependent activator of IFN-regulatory factors CD11c cells for the expression of proinflammatory (DAI), interferon-activated gene 204 (Ifi204, IFI16 in humans), cytokines and the recruitment of inflammatory mono- and cyclic-GMP-AMP (cGAMP) synthase (cGAS, encoded by cytes (iMos) to the draining lymph node (dLN). In the M21d1) respond to cytosolic DNA (Wu and Chen, 2014). dLN, the major producers of IFN-I were infected To signal, PRRs use adapters. The adapters for TLRs are MyD88 iMos, which used the DNA sensor-adaptor STING and TIR-domain-containing adaptor-inducing IFN-b (TRIF); for to activate IRF7 and nuclear factor kB (NF-kB) RLRs, mitochondrial antiviral-signaling protein (MAVS); and for signaling to induce the expression of IFN-a and DNA-sensing PRRs, STING (Wu and Chen, 2014). Additional IFN-b, respectively. Thus, in vivo, two pathways of signaling steps downstream of these adapters link PRRs to the DNA pathogen sensing act sequentially in two activation of several transcription factors, most frequently IRF3, IRF7, and NF-kB. These transcription factors induce the expres- distinct cell types to orchestrate resistance to a viral sion of IFN-I and pro-inflammatory cytokines (Wu and Chen, disease. 2014). Soon after breaching epithelia, many pathogenic viruses INTRODUCTION rapidly spread through afferent lymphatics to the regional drain- ing lymph nodes (dLNs), from where they disseminate through The ability of the innate immune system to sense infection is efferent lymphatics to the bloodstream, ultimately reaching their essential to mount innate and adaptive immune responses (Iwa- target organs (Flint et al., 2009; Virgin, 2007). Ectromelia virus, an saki and Medzhitov, 2010; Wu and Chen, 2014). It is well estab- Orthopoxvirus (a genus of large DNA viruses), is the causative lished that pathogen recognition receptors (PRRs) recognize agent of mousepox, the mouse homolog of human smallpox (Es- pathogen-associated molecular patterns (PAMPs) to activate teban and Buller, 2005). After infection through the skin of the innate immune signaling pathways (Iwasaki and Medzhitov, footpad, ECTV spreads lympho-hematogenously to cause sys- 2010). PAMPs are typically microbial nucleic acids and other temic disease. Indeed, ECTV was the virus used to describe macromolecules with repetitive structures common to patho- this form of dissemination (Fenner, 1948) and is used as the text- gens but not usually encountered in uninfected cells (Schenten book paradigm of lympho-hematogenous spread (Flint et al., and Medzhitov, 2011). PRR-initiated signaling cascades after 2009; Virgin, 2007). virus infection culminate in the expression of type I interferon During lympho-hematogenous dissemination, a swift anti-viral (IFN-I), pro-inflammatory cytokines, and chemokines that recruit innate response in the dLN can play a major role in restricting 1148 Immunity 43, 1148–1159, December 15, 2015 ª2015 Elsevier Inc. A B C B6 Tmem173gt Myd88-/- Mavs-/- 100 10 **** B6 **** 80 gt ** 8 Tmem173 H&E 60 Myd88-/- ** 6 -/- 40 Mavs 4 Survival (%) 20 2 Anti-ECTV 0 PFU /100 mg (Log10)0 0102030 gt /- dpi B6 - -/- Myd88 Mavs Tmem173 D E F IFNβ IFNα5 150 IFNβ 150 IFNα5 100 100 ** ** B6 ** ** 80 ** 80 -/- 100 * 100 * Tlr9 60 60 Tlr2-/- 40 -/- 40 50 50 Il1r Survival (%) Zbp1 -/- 20 to Naive Relative 20 Relative to Naive Relative 0 0 0 0 t - -/- -/- gt -/- -/- g -/- -/ 0102030 B6 B6 B6 B6 r9 dpi Tlr9 Tl Mavs Mavs Myd88 Myd88 Tmem173 Tmem173 Figure 1. TLR9, MyD88, and STING Are Critical for Resistance to Mousepox and the Efficient Induction of IFN-I in Lymph Nodes Mice were infected with 3,000 PFUs of ECTV in the footpad. (A) Survival of the indicated mice. (B) Virus loads in the livers of the indicated mice at 7 dpi as determined by plaque assay. (C) Liver sections of the indicated mice at 7 dpi stained with H&E (top) or immunostained with anti-ECTV Ab (bottom). (D) Expression of IFN-I in the dLNs of the indicated mice at 2.5 dpi as determined by RT-qPCR. (E) Survival of the indicated mice. (F) Expression of IFN-I in the dLNs of the indicated mice at 2.5 dpi as determined by RT-qPCR. Data are shown either as individual mice with mean ± SEM or as mean ± SEM. Each panel displays data from one experiment with five mice per group and is representative of three similar experiments. For all, *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. viral spread and deterring disease (Fang et al., 2008; Junt et al., well as of MAVS- and STING-driven pathways, in resistance 2007; Kastenmu¨ ller et al., 2012). It is therefore important to un- to mousepox and IFN-I expression in vivo during acute ECTV derstand how different mechanisms of virus sensing contribute infection. We found that after infection with ECTV in the footpad, to this process. Here we dissect the contribution of PRRs to mice deficient in MyD88 (Myd88À/À) or with inactive STING the antiviral response to ECTV in the dLN. We show that the (Tmem173gt), but not those lacking MAVS (MavsÀ/À), were sus- TLR9-MyD88-IRF7 axis is necessary in CD11c+ cells for the che- ceptible to lethal mousepox (Figure 1A). Nevertheless, death mokine-driven recruitment of inflammatory monocytes (iMos) to occurred in 100% of Myd88À/À but in only 80% Tmem173gt the dLN, but not directly essential for IFN-I production. Once mice, a difference that was reproducible and highly significant iMos are infected, they use STING-IRF7 and STING-NF-kB path- (p < 0.0001) suggesting a more profound impairment in the ways to induce the expression of IFN-a and IFN-b, respectively. absence of MyD88 than in the absence of STING signaling. Collectively, our work shows that the TLR9-MyD88-IRF7 and Consistently, virus loads (Figure 1B) and pathology (Figure 1C) STING-IRF7 or STING-NF-kB pathways have non-redundant, in the livers of Myd88À/À and Tmem173gt, but not of MavsÀ/À, complementary, and sequential roles in IFN-I expression in the mice were significantly higher than in B6 mice. Notably, the virus dLNs and in resistance to a highly pathogenic viral disease. titers were significantly lower (p < 0.01) and the liver pathology was somewhat milder in Tmem173gt than in Myd88À/À mice. RESULTS We next performed reverse transcriptase quantitative polymer- ase chain reaction (RT-qPCR) on RNA obtained from the dLNs TLR9-MyD88 and STING Are Critical for Resistance to at 2.5 days post infection (dpi). We found that Myd88À/À and Mousepox and the Efficient Induction of IFN-I in Lymph Tmem173gt but not MavsÀ/À mice expressed significantly lower Nodes levels of ‘‘early’’ IFN-b and ‘‘late’’ IFN-a5 than B6 mice.
Load more

Recommended publications
  • Ectromelia Virus (Mousepox)
    technical sheet Ectromelia Virus (Mousepox) Classification the virus its name, ectromelia, or partial amputation of DNA virus, enveloped the limbs and tail. Family Diagnosis Poxviridae Mousepox should be suspected if animals with the above clinical signs are seen in the animal facility, or Affected species there is unexplained widespread mortality in susceptible strains. Serologic diagnosis through MFIA™/ELISA Laboratory mice, wild mice, and other wild rodents or IFA is possible; if animals recover, they produce protective antibodies. Lesions strongly suggestive of Frequency mousepox are noted on necropsy, including splenic Rare in laboratory mice, uncommon in wild mice. fibrosis in recovered animals, and liver, spleen, and skin lesions in ill animals. Histologically, intracytoplasmic Transmission inclusion bodies are seen in skin lesions. PCR of skin Ectromelia virus, which causes the disease mousepox, lesions can be used for confirmation. Vaccination with is transmitted by direct contact or by fomites. Although vaccinia virus as part of an experimental protocol may many routes of infection are possible experimentally, give false positive serology results. exposure to the virus via cutaneous trauma is the natural route of infection. Lesions appear 7-11 days Interference with Research after infection in susceptible strains, and virus is shed Overwhelming mortality (near 100%) in susceptible for 3 weeks. However, virus has been found in scabs strains may have a negative effect on research and feces for as long as 16 weeks post-infection. programs and animal facilities. Ectromelia virus infection Cage-to-cage transmission in mousepox infections is may modify the phagocytic response in resistant primarily through handling of infected mice.
    [Show full text]
  • Investigation of Poxvirus Host-Range and Gene Expression in Mammalian Cells
    ABSTRACT Title: INVESTIGATION OF POXVIRUS HOST-RANGE AND GENE EXPRESSION IN MAMMALIAN CELLS. Jorge David Méndez Ríos, Doctor of Philosophy, 2014. Directed By: Dr. Bernard Moss, Chief, Laboratory of Viral Diseases, NIAID, NIH; Adjunct Professor Department of Cell Biology and Molecular Genetics, University of Maryland; Members of the Poxviridae family have been known as human pathogens for centuries. Their impact in society included several epidemics that decimated the population. In the last few centuries, Smallpox was of great concern that led to the development of our modern vaccines. The systematic study of Poxvirus host-range and immunogenicity provided the knowledge to translate those observations into practice. After the global vaccination campaign by the World Health Organization, Smallpox was the first infectious disease to be eradicated. Nevertheless, diseases such as Monkeypox, Molluscum contagiosum, new bioterrorist threads, and the use of poxviruses as vaccines or vectors provided the necessity to further understand the host-range from a molecular level. Here, we take advantage of the newly developed technologies such as 454 pyrosequencing and RNA-Seq to address previously unresolved questions for the field. First, we were able to identify the Erytrhomelagia-related poxvirus (ERPV) 25 years after its isolation in Hubei, China. Whole-genome sequencing and bioinformatics identified ERPV as an Ectromelia strain closely related to the Ectromelia Naval strain. Second, by using RNA-Seq, the first MOCV in vivo and in vitro transcriptome was generated. New tools have been developed to support future research and for this human pathogen. Finally, deep-sequencing and comparative genomes of several recombinant MVAs (rMVAs) in conjunction with classical virology allowed us to confirm several genes (O1, F5, C17, F11) association to plaque formation in mammalian cell lines.
    [Show full text]
  • Control of Lymphocytic Choriomeningitis Virus Infection in Granzyme B Deficient Mice
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Virology 305, 1–9 (2003) doi:10.1006/viro.2002.1754 Control of Lymphocytic Choriomeningitis Virus Infection in Granzyme B Deficient Mice Allan J. Zajac,*,† John M. Dye,* and Daniel G. Quinn*,1 *Department of Microbiology and Immunology, Loyola University Chicago, Maywood, Illinois 60153; and †Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 35294 Received July 11, 2001; returned to author for revision October 5, 2001; accepted August 30, 2002 We have investigated whether granzyme B (GzmB)is required for effective cytotoxic T lymphocyte (CTL)mediated control of lymphocytic choriomeningitis virus (LCMV)infection. Clearance of LCMV from tissues of GzmB-deficient (GzmB Ϫ)mice following intraperitoneal infection with LCMV was impaired compared with control mice; however, the virus was ultimately eliminated. The impaired clearance of LCMV in GzmBϪ mice was not due to a deficiency in the generation of LCMV-specific T cells. In addition, CTL from LCMV-infected GzmBϪ mice efficiently lysed virus-infected cells in vitro, but were deficient in their ability to induce rapid DNA fragmentation in target cells. We examined whether the development of protective immunity against intracranial (i.c.)rechallenge with LCMV was compromised in GzmB Ϫ mice. We found that clearance of LCMV from the brain following secondary i.c. infection also was slower in the absence of GzmB; however, the virus was ultimately eliminated and the mice survived. Our data indicate that clearance of LCMV is delayed in the absence of GzmB expression, but that other CTL effector molecules can compensate for the absence of this granule constituent in vivo.
    [Show full text]
  • A System Based-Approach to Examine Cytokine Response in Poxvirus-Infected Macrophages
    viruses Article A System Based-Approach to Examine Cytokine Response in Poxvirus-Infected Macrophages Pui-San Wong 1,†, Richard Sutejo 2,†,‡, Hui Chen 2,§ , Sock-Hoon Ng 1, Richard J. Sugrue 2 and Boon-Huan Tan 1,3,* 1 Defence Medical and Environmental Research Institute, DSO National Labs, Singapore 117510, Singapore; [email protected] (P.-S.W.); [email protected] (S.-H.N.) 2 School of Biological Sciences, Nanyang Technological University, Singapore 637551, Singapore; [email protected] (R.S.); [email protected] (H.C.); [email protected] (R.J.S.) 3 Infection and Immunity, LKC School of Medicine, Nanyang Technological University, Singapore 308232, Singapore * Correspondence: [email protected]; Tel: +65-64857238 † These authors contributed equally to this work. ‡ Current address: International Institute for life Sciences, Jakarta Timur 13210, Indonesia. Received: 9 October 2018; Accepted: 30 November 2018; Published: 5 December 2018 Abstract: The poxviruses are large, linear, double-stranded DNA viruses about 130 to 230 kbp, that have an animal origin and evolved to infect a wide host range. Variola virus (VARV), the causative agent of smallpox, is a poxvirus that infects only humans, but other poxviruses such as monkey poxvirus and cowpox virus (CPXV) have crossed over from animals to infect humans. Therefore understanding the biology of poxviruses can devise antiviral strategies to prevent these human infections. In this study we used a system-based approach to examine the host responses to three orthopoxviruses, CPXV, vaccinia virus (VACV), and ectromelia virus (ECTV) in the murine macrophage RAW 264.7 cell line.
    [Show full text]
  • Enhanced Resistance in STAT6-Deficient Mice to Infection with Ectromelia Virus
    Enhanced resistance in STAT6-deficient mice to infection with ectromelia virus Surendran Mahalingam*†, Gunasegaran Karupiah‡, Kiyoshi Takeda§, Shizuo Akira¶, Klaus I. Matthaei*, and Paul S. Foster* *Division of Biochemistry and Molecular Biology, The John Curtin School of Medical Research, Australian National University, Canberra ACT 2601, Australia; ‡Department of Pathology, Division of Faculty of Medicine, Blackburn Building, D06, University of Sydney, Sydney NSW 2006, Australia; §Department of Host Defense, Research Institute for Microbial Disease, Osaka University, Suita-shi, Osaka 565-0871, Japan; and ¶Department of Biochemistry, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya, Hyogo 663-8501, Japan Communicated by Frank J. Fenner, Australian National University, Canberra, Australia, March 27, 2001 (received for review January 25, 2001) -We inoculated BALB͞c mice deficient in STAT6 (STAT6؊/؊) and their characterized in Leishmania major infection. Studies have indi wild-type (wt) littermates (STAT6؉/؉) with the natural mouse cated that control of lesion growth induced by L. major in pathogen, ectromelia virus (EV). STAT6؊/؊ mice exhibited in- genetically resistant mice (C57BL͞6) is associated with the creased resistance to generalized infection with EV when com- expansion of CD4ϩ T helper 1 (Th1) cells and the production of ؉ ؉ pared with STAT6 / mice. In the spleens and lymph nodes of cytokines such as IL-12, IFN-␥, and IL-2 (13, 14). By contrast, ؊ ؊ STAT6 / mice, T helper 1 (Th1) cytokines were induced at earlier nonhealing responses in susceptible mice (BALB͞c) have been time points and at higher levels postinfection when compared with related to the expansion of CD4ϩ Th2 cells and the production ؉ ؉ those in STAT6 / mice.
    [Show full text]
  • Ectromelia Virus Disease Characterization in the BALB/C Mouse: a Surrogate Model for Assessment of Smallpox Medical Countermeasures
    viruses Article Ectromelia Virus Disease Characterization in the BALB/c Mouse: A Surrogate Model for Assessment of Smallpox Medical Countermeasures Jennifer Garver 1,*, Lauren Weber 1, Eric M. Vela 2, Mike Anderson 1, Richard Warren 3, Michael Merchlinsky 3, Christopher Houchens 3 and James V. Rogers 1 1 Battelle Biomedical Research Center, West Jefferson, OH 43162, USA; [email protected] (L.W.); [email protected] (M.A.); [email protected] (J.V.R.) 2 Vaccine and Gene Therapy Institute, Oregon Health and Science University, Portland, OR 97239, USA; [email protected] 3 Biomedical Advanced Research and Development Authority, US Department of Health and Human Services, Washington, DC 20201, USA; [email protected] (R.W.); [email protected] (M.M.); [email protected] (C.H.) * Correspondence: [email protected]; Tel.: +1-614-424-3984; Fax: +1-614-458-3984 Academic Editor: Curt Hagedorn Received: 22 June 2016; Accepted: 19 July 2016; Published: 22 July 2016 Abstract: In 2007, the United States’ Food and Drug Administration (FDA) issued guidance concerning animal models for testing the efficacy of medical countermeasures against variola virus (VARV), the etiologic agent for smallpox. Ectromelia virus (ECTV) is naturally-occurring and responsible for severe mortality and morbidity as a result of mousepox disease in the murine model, displaying similarities to variola infection in humans. Due to the increased need of acceptable surrogate animal models for poxvirus disease, we have characterized ECTV infection in the BALB/c mouse. Mice were inoculated intranasally with a high lethal dose (125 PFU) of ECTV, resulting in complete mortality 10 days after infection.
    [Show full text]
  • New Insights Into the Evolutionary and Genomic Landscape of Molluscum Contagiosum Virus (MCV) Based on Nine MCV1 and Six MCV2 Complete Genome Sequences
    viruses Article New Insights into the Evolutionary and Genomic Landscape of Molluscum Contagiosum Virus (MCV) based on Nine MCV1 and Six MCV2 Complete Genome Sequences Tomaž M. Zorec 1 , Denis Kutnjak 2 , Lea Hošnjak 1 , Blanka Kušar 1, Katarina Trˇcko 3, Boštjan J. Kocjan 1 , Yu Li 4, Miljenko Križmari´c 5, Jovan Miljkovi´c 5, Maja Ravnikar 2 and Mario Poljak 1,* 1 Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Zaloška 4, SI-1000 Ljubljana, Slovenia; [email protected] (T.M.Z.); [email protected] (L.H.); [email protected] (B.K.); [email protected] (B.J.K.) 2 Department of Biotechnology and Systems Biology, National Institute of Biology, Veˇcna pot 111, SI-1000 Ljubljana, Slovenia; [email protected] (D.K.); [email protected] (M.R.) 3 Department of Dermatovenereology, University Medical Centre Maribor, Ljubljanska ulica 5, SI-2000 Maribor, Slovenia; [email protected] 4 Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road NE, Atlanta, GA 30333, USA; [email protected] 5 Faculty of Medicine, University of Maribor, Taborska Ulica 6b, SI-2000 Maribor, Slovenia; [email protected] (M.K.); [email protected] (J.M.) * Correspondence: [email protected]; Tel.: +386-1-543-7415 Received: 9 October 2018; Accepted: 25 October 2018; Published: 26 October 2018 Abstract: Molluscum contagiosum virus (MCV) is the sole member of the Molluscipoxvirus genus and the causative agent of molluscum contagiosum (MC), a common skin disease.
    [Show full text]
  • Evaluation of Taterapox Virus in Small Animals
    viruses Article Evaluation of Taterapox Virus in Small Animals Scott Parker * ID , Ryan Crump, Hollyce Hartzler and R. Mark Buller † Department of Molecular Microbiology and Immunology, Saint Louis University School of Medicine, 1100 South Grand Boulevard, St. Louis, MO 63104, USA; [email protected] (R.C.); [email protected] (H.H.); [email protected] (R.M.B.) * Correspondence: [email protected]; Tel.: +1-314-783-6107 † Deceased. Academic Editors: Hermann Meyer, Jônatas Abrahão and Erna Geessien Kroon Received: 8 July 2017; Accepted: 24 July 2017; Published: 1 August 2017 Abstract: Taterapox virus (TATV), which was isolated from an African gerbil (Tatera kempi) in 1975, is the most closely related virus to variola; however, only the original report has examined its virology. We have evaluated the tropism of TATV in vivo in small animals. We found that TATV does not infect Graphiurus kelleni, a species of African dormouse, but does induce seroconversion in the Mongolian gerbil (Meriones unguiculatus) and in mice; however, in wild-type mice and gerbils, the virus produces an unapparent infection. Following intranasal and footpad inoculations with 1 × 106 plaque forming units (PFU) of TATV, immunocompromised stat1−/− mice showed signs of disease but did not die; however, SCID mice were susceptible to intranasal and footpad infections with 100% mortality observed by Day 35 and Day 54, respectively. We show that death is unlikely to be a result of the virus mutating to have increased virulence and that SCID mice are capable of transmitting TATV to C57BL/6 and C57BL/6 stat1−/− animals; however, transmission did not occur from TATV inoculated wild-type or stat1−/− mice.
    [Show full text]
  • Mousepox Resulting from Use of Ectromelia Virus-Contaminated, Imported Mouse Serum
    Comparative Medicine Vol 50, No 4 Copyright 2000 August 2000 by the American Association for Laboratory Animal Science Mousepox Resulting from Use of Ectromelia Virus-Contaminated, Imported Mouse Serum Neil S. Lipman,1* Scott Perkins,1 Hai Nguyen,1 Martin Pfeffer,2 and Hermann Meyer3 Abstract Mousepox was identified in a single mouse-holding room in early 1999 after a group of 20 CAF1/Hsd mice were inoculated SC with a killed murine spindle cell tumor line, S1509A. The cell line had been used without complications multiple times and was determined to be free of viral contamination on the basis of results of mouse antibody production testing. Of the 20 mice inoculated, 12 mice died by postinoculation day 8. Severe lymphoid and hepatic necrosis was observed in select mice subjected to histologic examination. Ballooning degeneration of epi- thelial cells with intracytoplasmic eosinophilic inclusion bodies was observed in the skin overlying the inoculation site of the single mouse from which this tissue site was evaluated. Presence of ectromelia virus was confirmed by use of immunohistochemical and polymerase chain reaction analyses, and the virus was isolated after serum, pooled from 5 of the index cases, was inoculated into an immune-naive mouse. Investigation into the source of virus con- tamination included inoculating mice with aliquots of various S1509A freeze dates; chemically defined media and supplements, including fetal bovine serum; and two lots of pooled commercial mouse sera, after heat inactivation at 56ЊC for 30 minutes used as a medium supplement. One lot of pooled commercial mouse serum was identified as the source of ectromelia virus.
    [Show full text]
  • Granzyme a Is Critical for Recovery of Mice from Infection With
    Proc. Natl. Acad. Sci. USA Vol. 93, pp. 5783-5787, June 1996 Immunology Granzyme A is critical for recovery of mice from infection with the natural cytopathic viral pathogen, ectromelia (poxvirus/cytolytic leukocytes/proteases) ARNO MULLBACHER*, KLAUS EBNETtS, ROBERT V. BLANDEN*, RON THA HLA*, THOMAS STEHLEt, CRISAN MUSETEANUt, AND MARKUS M. SIMONt§ *Division of Immunology and Cell Biology, John Curtin School of Medical Research, The Australian National University, Canberra City, Australian Capital Territory 2601, Australia; and tMax-Planck-Institut for Immunbiology, Stiibeweg 51, D-79108 Freiburg, Germany Communicated by Frank Fenner, The John Curtin School of Medical Research, The Australian National University, Canberra, Australia, February 7, 1996 (received for review December 6, 1995) ABSTRACT Cytolytic lymphocytes are of cardinal impor- normal littermates between 6 and 12 weeks of age were used tance in the recovery from primary viral infections. Both throughout the experiments. natural killer cells and cytolytic T cells mediate at least part Viruses. The virulent Moscow strain ectromelia virus was of their effector function by target cell lysis and DNA frag- grown in mouse spleen, the intracellular naked ectromelia mentation. Two proteins, perforin and granzyme B, contained virus (INV), and the extracellular enveloped ectromelia virus within the cytoplasmic granules ofthese cytolytic effector cells (EEV) in tissue culture, as has been described previously (12, have been shown to be directly involved in these processes. A 13). Viruses were purified and titrated on BS-C-1 monkey cell third protein contained within these granules, granzyme A, monolayers as has been described (14). The predominance of has so far not been attributed with any biological relevance.
    [Show full text]
  • Structure-Function Studies of the Vaccinia Virus Complement Control Protein
    University of Louisville ThinkIR: The University of Louisville's Institutional Repository Electronic Theses and Dissertations 8-2002 Structure-function studies of the vaccinia virus complement control protein. Scott Alan Smith University of Louisville Follow this and additional works at: https://ir.library.louisville.edu/etd Recommended Citation Smith, Scott Alan, "Structure-function studies of the vaccinia virus complement control protein." (2002). Electronic Theses and Dissertations. Paper 1347. https://doi.org/10.18297/etd/1347 This Doctoral Dissertation is brought to you for free and open access by ThinkIR: The University of Louisville's Institutional Repository. It has been accepted for inclusion in Electronic Theses and Dissertations by an authorized administrator of ThinkIR: The University of Louisville's Institutional Repository. This title appears here courtesy of the author, who has retained all other copyrights. For more information, please contact [email protected]. STRUCTURE-FUNCTION STUDIES OF THE VACCINIA VIRUS COMPLEMENT CONTROL PROTEIN By Scott Alan Smith A.S. Community College of Allegheny County, 1995 B.S. California University of Pennsylvania, 1997 A Dissertation Submitted to the Faculty of the Graduate School of the University of Louisville in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy Department of Microbiology and Immunology University of Louisville Louisville, Kentucky August 2002 STUCTURE-FUNCTION STUDIES OF THE VACCINIA VIRUS COMPLEMENT CONTROL PROTEIN By Scott Alan Smith
    [Show full text]
  • Loss of Actin-Based Motility Impairs Ectromelia Virus Release in Vitro but Is Not Critical to Spread in Vivo
    viruses Article Loss of Actin-Based Motility Impairs Ectromelia Virus Release In Vitro but Is Not Critical to Spread In Vivo Melanie Laura Duncan 1, Jacquelyn Horsington 1, Preethi Eldi 2,3, Zahrah Al Rumaih 2,3, Gunasegaran Karupiah 2,3 and Timothy P. Newsome 1,* ID 1 School of Life and Environmental Sciences, The University of Sydney, Sydney, NSW 2006, Australia; [email protected] (M.L.D.); [email protected] (J.H.) 2 School of Medicine, College of Health and Medicine, The University of Tasmania, Hobart, TAS 7005, Australia; [email protected] (P.E.); [email protected] (Z.A.R.); [email protected] (G.K.) 3 The John Curtin School of Medical Research, Australian National University, Canberra, ACT 2601, Australia * Correspondence: [email protected]; Tel.: +61-2-9351-2901 Received: 15 February 2018; Accepted: 1 March 2018; Published: 5 March 2018 Abstract: Ectromelia virus (ECTV) is an orthopoxvirus and the causative agent of mousepox. Like other poxviruses such as variola virus (agent of smallpox), monkeypox virus and vaccinia virus (the live vaccine for smallpox), ECTV promotes actin-nucleation at the surface of infected cells during virus release. Homologs of the viral protein A36 mediate this function through phosphorylation of one or two tyrosine residues that ultimately recruit the cellular Arp2/3 actin-nucleating complex. A36 also functions in the intracellular trafficking of virus mediated by kinesin-1. Here, we describe the generation of a recombinant ECTV that is specifically disrupted in actin-based motility allowing us to examine the role of this transport step in vivo for the first time.
    [Show full text]