UNC-Chapel Hill Coronavirus Research
User Collection Public
A collection of articles and student papers related to coronaviruses that have been authored by researchers at the University of North Carolina at Chapel Hill.
Works (1049)
981. Systematic Assembly of a Full-Length Infectious Clone of Human Coronavirus NL63
- Title Tesim:
- Systematic Assembly of a Full-Length Infectious Clone of Human Coronavirus NL63
- Creator:
- Yount, Boyd, Pickles, Raymond J., Donaldson, Eric F., Baric, Ralph S., Burkett, Susan, and Sims, Amy C.
- Date of publication:
- 2008
- Abstract Tesim:
- Historically, coronaviruses were predominantly associated with mild upper respiratory disease in humans. More recently, three novel coronaviruses associated with severe human respiratory disease were found, including (i) the severe acute respiratory syndrome coronavirus, associated with a significant atypical pneumonia and 10% mortality; (ii) HKU-1, associated with chronic pulmonary disease; and (iii) NL63, associated with both upper and lower respiratory tract disease in children and adults worldwide. These discoveries establish coronaviruses as important human pathogens and underscore the need for continued research toward the development of platforms that will enable genetic manipulation of the viral genome, allowing for rapid and rational development and testing of candidate vaccines, vaccine vectors, and therapeutics. In this report, we describe a reverse genetics system for NL63, whereby five contiguous cDNAs that span the entire genome were used to generate a full-length cDNA. Recombinant NL63 viruses which contained the expected marker mutations replicated as efficiently as the wild-type NL63 virus. In addition, we engineered the heterologous green fluorescent protein gene in place of open reading frame 3 (ORF3) of the NL63 clone, simultaneously creating a unique marker for NL63 infection and demonstrating that the ORF3 protein product is nonessential for the replication of NL63 in cell culture. The availability of the NL63 and NL63gfp clones and recombinant viruses provides powerful tools that will help advance our understanding of this important human pathogen.
- Resource type:
- Article
- Admin Note:
- deposited as part of the 1Science batch
- Affiliation Label Tesim:
- Department of Epidemiology, Department of Microbiology and Immunology, and Cystic Fibrosis and Pulmonary Diseases Research and Treatment Center
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/5722-bx98
- Identifier:
- https://dx.doi.org/10.1128/JVI.01804-08
- ISSN:
- 0022-538X
- Journal Issue:
- 23
- Journal Title:
- Journal of Virology
- Journal Volume:
- 82
- Keyword:
- Macaca mulatta, RNA, Viral, coronavirus, Blotting, Western, Virus Assembly, Bronchi, Mutation, Animals, Virus Replication, Respiratory Tract Infections, Recombination, Genetic, Humans, and Open Reading Frames
- Language Label:
- English
- ORCID:
- Other Affiliation:
- Page End:
- 11957
- Page Start:
- 11948
- Person:
- Yount, Boyd, Pickles, Raymond J., Donaldson, Eric F., Baric, Ralph S., Burkett, Susan, and Sims, Amy C.
- Rights Statement Label:
- In Copyright
- Source:
- 9019s674n
982. Structural Basis for Potent Cross-Neutralizing Human Monoclonal Antibody Protection against Lethal Human and Zoonotic Severe Acute Respiratory Syndrome Coronavirus Challenge
- Title Tesim:
- Structural Basis for Potent Cross-Neutralizing Human Monoclonal Antibody Protection against Lethal Human and Zoonotic Severe Acute Respiratory Syndrome Coronavirus Challenge
- Creator:
- Sheahan, Timothy, Donaldson, Eric, Stadler, Konrad, Baric, Ralph, Rockx, Barry, Corti, Davide, and Lanzavecchia, Antonio
- Date of publication:
- 2008
- Abstract Tesim:
- Severe acute respiratory syndrome coronavirus (SARS-CoV) emerged in 2002, and detailed phylogenetic and epidemiological analyses have suggested that it originated from animals. The spike (S) glycoprotein has been identified as a major component of protective immunity, and 23 different amino acid changes were noted during the expanding epidemic. Using a panel of SARS-CoV recombinants bearing the S glycoproteins from isolates representing the zoonotic and human early, middle, and late phases of the epidemic, we identified 23 monoclonal antibodies (MAbs) with neutralizing activity against one or multiple SARS-CoV spike variants and determined the presence of at least six distinct neutralizing profiles in the SARS-CoV S glycoprotein. Four of these MAbs showed cross-neutralizing activity against all human and zoonotic S variants in vitro, and at least three of these were mapped in distinct epitopes using escape mutants, structure analyses, and competition assays. These three MAbs (S109.8, S227.14, and S230.15) were tested for use in passive vaccination studies using lethal SARS-CoV challenge models for young and senescent mice with four different homologous and heterologous SARS-CoV S variants. Both S227.14 and S230.15 completely protected young and old mice from weight loss and virus replication in the lungs for all viruses tested, while S109.8 completely protected mice from weight loss and clinical signs in the presence of viral titers. We conclude that a single human MAb can confer broad protection against lethal challenge with multiple zoonotic and human SARS-CoV isolates, and we identify a robust cocktail formulation that targets distinct epitopes and minimizes the likely generation of escape mutants.
- Resource type:
- Article
- Admin Note:
- deposited as part of the 1Science batch
- Affiliation Label Tesim:
- Department of Microbiology and Immunology and Department of Epidemiology
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/5v69-2w75
- Identifier:
- https://dx.doi.org/10.1128/JVI.02377-07
- ISSN:
- 0022-538X
- Journal Issue:
- 7
- Journal Title:
- Journal of Virology
- Journal Volume:
- 82
- Keyword:
- neutralizing antibody, human, Cross protection, Acute, Monoclonal antibody, Virology, Virus, and coronavirus
- Language Label:
- English
- ORCID:
- Other Affiliation:
- , Novartis Vaccines, and Institute for Research in Biomedicine
- Page End:
- 3235
- Page Start:
- 3220
- Person:
- Sheahan, Timothy, Donaldson, Eric, Stadler, Konrad, Baric, Ralph, Rockx, Barry, Corti, Davide, and Lanzavecchia, Antonio
- Rights Statement Label:
- In Copyright
- Source:
- 6108vg687
983. Persistent Replication of Severe Acute Respiratory Syndrome Coronavirus in Human Tubular Kidney Cells Selects for Adaptive Mutations in the Membrane Protein
- Title Tesim:
- Persistent Replication of Severe Acute Respiratory Syndrome Coronavirus in Human Tubular Kidney Cells Selects for Adaptive Mutations in the Membrane Protein
- Creator:
- Clementi, Massimo, Sims, Amy, Sampaolo, Michela, Conaldi, Pier Giulio, Pacciarini, Filippo, Poli, Guido, Ghezzi, Silvia, Baric, Ralph, Canducci, Filippo, Ferioli, Elena, and Vincenzi, Elisa
- Date of publication:
- 2008
- Abstract Tesim:
- Severe acute respiratory syndrome (SARS) is a systemic disease characterized by both lung pathology and widespread extrapulmonary virus dissemination causing multiple organ injuries. In this regard, renal dysfunction is an ominous sign in patients with SARS. Indeed, clusters of SARS coronavirus (SARS-CoV) particles have been detected in the cytoplasm of renal tubular epithelial cells in postmortem studies, explaining the presence of infectious virus in the urine of SARS patients. In order to investigate the potential SARS-CoV kidney tropism, we have evaluated the susceptibility of human renal cells of tubular and glomerular origin to in vitro SARS-CoV infection. Immortalized cultures of differentiated proximal tubular epithelial cells (PTEC), glomerular mesangial cells (MC), and glomerular epithelial cells (podocytes) were found to express the SARS-CoV receptor angiotensin-converting enzyme 2 on their surface. Productive infection, however, occurred only in PTEC but not in glomerular cells. A transient infection with poor virus production was observed in MC, whereas podocytes were not permissive to SARS-CoV infection. In contrast to the cytopathic infection of the Vero E6 cell line, SARS-CoV did not cause overt cytopathic effects in PTEC or MC. Of interest, PTEC, but not MC, maintained stable levels of SARS-CoV production in serial subcultures, suggesting a persistent state of infection. In this regard, a SARS-CoV variant with increased replication capacity in PTEC was selected after four serial subculture passages. This SARS-CoV variant acquired a single nonconservative amino acid change from glutamic acid (E) to alanine (A) at position 11 in the viral membrane (M) protein. The E11A point mutation was sufficient for enhanced SARS-CoV replication and persistence in PTEC when introduced in a SARS-CoV recombinant infectious clone. These findings indicate that human PTEC may represent a site of SARS-CoV productive and persistent replication favoring the emergence of viral variants with increased replication capacity, at least in these kidney cells.
- Resource type:
- Article
- Admin Note:
- deposited in the 1Science batch
- Affiliation Label Tesim:
- University of North Carolina at Chapel Hill
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/dajp-3m63
- Identifier:
- https://dx.doi.org/10.1128/JVI.00096-08
- ISSN:
- 0022-538X
- Journal Issue:
- 11
- Journal Title:
- Journal of Virology
- Journal Volume:
- 82
- Keyword:
- Viral disease, Respiratory disease, Severe acute respiratory syndrome, Mutation, Virology, Kidney, Membrane protein, Replication, Lung disease, Virus, Human, Infection, In vitro, and Coronavirus
- Language Label:
- English
- ORCID:
- Other Affiliation:
- San Raffaele Scientific Institute, , University of Pittsburgh Medical Center—Italy, and University of Insubria
- Page End:
- 5144
- Page Start:
- 5137
- Person:
- Clementi, Massimo, Sims, Amy, Sampaolo, Michela, Conaldi, Pier Giulio, Pacciarini, Filippo, Poli, Guido, Ghezzi, Silvia, Baric, Ralph, Canducci, Filippo, Ferioli, Elena, and Vincenzi, Elisa
- Rights Statement Label:
- In Copyright
- Source:
- 9593v104m
984. Pathways of Cross-Species Transmission of Synthetically Reconstructed Zoonotic Severe Acute Respiratory Syndrome Coronavirus
- Title Tesim:
- Pathways of Cross-Species Transmission of Synthetically Reconstructed Zoonotic Severe Acute Respiratory Syndrome Coronavirus
- Creator:
- Sheahan, Timothy, Baric, Ralph, Rockx, Barry, Corti, Davide, and Donaldson, Eric
- Date of publication:
- 2008
- Abstract Tesim:
- Zoonotic severe acute respiratory syndrome coronavirus (SARS-CoV) likely evolved to infect humans by a series of transmission events between humans and animals in markets in China. Virus sequence data suggest that the palm civet served as an amplification host in which civet and human interaction fostered the evolution of the epidemic SARS Urbani strain. The prototypic civet strain of SARS-CoV, SZ16, was isolated from a palm civet but has not been successfully cultured in vitro. To propagate a chimeric recombinant SARS-CoV bearing an SZ16 spike (S) glycoprotein (icSZ16-S), we constructed cell lines expressing the civet ortholog (DBT-cACE2) of the SARS-CoV receptor (hACE2). Zoonotic SARS-CoV was completely dependent on ACE2 for entry. Urbani grew with similar kinetics in both the DBT-cACE2 and the DBT-hACE2 cells, while icSZ16-S only grew in DBT-cACE2 cells. The SZ16-S mutant viruses adapted to human airway epithelial cells and displayed enhanced affinity for hACE2 but exhibited severe growth defects in the DBT-cACE2 cells, suggesting that the evolutionary pathway that promoted efficient hACE2 interactions simultaneously abolished efficient cACE2 interactions. Structural modeling predicted two distinct biochemical interaction networks by which zoonotic receptor binding domain architecture can productively engage hACE2, but only the Urbani mutational repertoire promoted efficient usage of both hACE2 and cACE2 binding interfaces. Since dual species tropism was preserved in Urbani, it is likely that the virus evolved a high affinity for cACE2/hACE2 receptors through adaptation via repeated passages between human and civet hosts. Furthermore, zoonotic SARS-CoV was variably neutralized by antibodies that were effective against the epidemic strain, highlighting their utility for evaluating passive immunization efficacy.
- Resource type:
- Article
- Admin Note:
- deposited as part of the 1Science batch
- Affiliation Label Tesim:
- Department of Microbiology and Immunology and Department of Epidemiology
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/act3-hb68
- Identifier:
- https://dx.doi.org/10.1128/JVI.00818-08
- ISSN:
- 0022-538X
- Journal Issue:
- 17
- Journal Title:
- Journal of Virology
- Journal Volume:
- 82
- Keyword:
- Severe acute respiratory syndrome, Virus, Transmission, Infection, Species, Coronavirus, Virology, Viral disease, Respiratory disease, and Lung disease
- Language Label:
- English
- ORCID:
- Other Affiliation:
- and Institute for Research in Biomedicine
- Page End:
- 8732
- Page Start:
- 8721
- Person:
- Sheahan, Timothy, Baric, Ralph, Rockx, Barry, Corti, Davide, and Donaldson, Eric
- Rights Statement Label:
- In Copyright
- Source:
- cj82kd03t
985. Mechanisms of Zoonotic Severe Acute Respiratory Syndrome Coronavirus Host Range Expansion in Human Airway Epithelium
- Title Tesim:
- Mechanisms of Zoonotic Severe Acute Respiratory Syndrome Coronavirus Host Range Expansion in Human Airway Epithelium
- Creator:
- Sheahan, Timothy, Rockx, Barry, Corti, Davide, Sims, Amy, Baric, Ralph, Pickles, Raymond, and Donaldson, Eric
- Date of publication:
- 2008
- Abstract Tesim:
- In 2003, severe acute respiratory syndrome coronavirus (SARS-CoV) emerged and caused over 8,000 human cases of infection and more than 700 deaths worldwide. Zoonotic SARS-CoV likely evolved to infect humans by a series of transmission events between humans and animals for sale in China. Using synthetic biology, we engineered the spike protein (S) from a civet strain, SZ16, into our epidemic strain infectious clone, creating the chimeric virus icSZ16-S, which was infectious but yielded progeny viruses incapable of propagating in vitro. After introducing a K479N mutation within the S receptor binding domain (RBD) of SZ16, the recombinant virus (icSZ16-S K479N) replicated in Vero cells but was severely debilitated in growth. The in vitro evolution of icSZ16-S K479N on human airway epithelial (HAE) cells produced two viruses (icSZ16-S K479N D8 and D22) with enhanced growth on HAE cells and on delayed brain tumor cells expressing the SARS-CoV receptor, human angiotensin I converting enzyme 2 (hACE2). The icSZ16-S K479N D8 and D22 virus RBDs contained mutations in ACE2 contact residues, Y442F and L472F, that remodeled S interactions with hACE2. Further, these viruses were neutralized by a human monoclonal antibody (MAb), S230.15, but the parent icSZ16-S K479N strain was eight times more resistant than the mutants. These data suggest that the human adaptation of zoonotic SARS-CoV strains may select for some variants that are highly susceptible to select MAbs that bind to RBDs. The epidemic, icSZ16-S K479N, and icSZ16-S K479N D22 viruses replicate similarly in the BALB/c mouse lung, highlighting the potential use of these zoonotic spike SARS-CoVs to assess vaccine or serotherapy efficacy in vivo.
- Resource type:
- Article
- Admin Note:
- deposited as part of the 1Science upload
- Affiliation Label Tesim:
- Department of Microbiology and Immunology and Department of Epidemiology
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/anq3-yj27
- Identifier:
- https://dx.doi.org/10.1128/JVI.02041-07
- ISSN:
- 0022-538X
- Journal Issue:
- 5
- Journal Title:
- Journal of Virology
- Journal Volume:
- 82
- Keyword:
- Epithelium, Acute, Respiratory tract, coronavirus, Mechanism of action, Virology, Virus, Host range, and Human
- Language Label:
- English
- ORCID:
- Other Affiliation:
- and Institute for Research in Biomedicine
- Page End:
- 2285
- Page Start:
- 2274
- Person:
- Sheahan, Timothy, Rockx, Barry, Corti, Davide, Sims, Amy, Baric, Ralph, Pickles, Raymond, and Donaldson, Eric
- Rights Statement Label:
- In Copyright
- Source:
- bv73c4946
986. mRNA Display Design of Fibronectin-based Intrabodies That Detect and Inhibit Severe Acute Respiratory Syndrome Coronavirus Nucleocapsid Protein
- Title Tesim:
- mRNA Display Design of Fibronectin-based Intrabodies That Detect and Inhibit Severe Acute Respiratory Syndrome Coronavirus Nucleocapsid Protein
- Creator:
- Deng, Hongyu, Sun, Ren, Liao, Hsiang-I, Roberts, Richard W., Olson, C. Anders, Wong, Elaine, Hwang, Seungmin, and Baric, Ralph S.
- Date of publication:
- 2009
- Abstract Tesim:
- The nucleocapsid (N) protein of severe acute respiratory syndrome (SARS) coronavirus plays important roles in both viral replication and modulation of host cell processes. New ligands that target the N protein may thus provide tools to track the protein inside cells, detect interaction hot spots on the protein surface, and discover sites that could be used to develop new anti-SARS therapies. Using mRNA display selection and directed evolution, we designed novel antibody-like protein affinity reagents that target SARS N protein with high affinity and selectivity. Our libraries were based on an 88-residue variant of the 10th fibronectin type III domain from human fibronectin (10Fn3). This selection resulted in eight independent 10Fn3 intrabodies, two that require the N-terminal domain for binding and six that recognize the C terminus, one with Kd = 1.7 nm. 10Fn3 intrabodies are well expressed in mammalian cells and are relocalized by N in SARS-infected cells. Seven of the selected intrabodies tested do not perturb cellular function when expressed singly in vivo and inhibit virus replication from 11- to 5900-fold when expressed in cells prior to infection. Targeting two sites on SARS-N simultaneously using two distinct 10Fn3s results in synergistic inhibition of virus replication.
- Resource type:
- Article
- Admin Note:
- deposited as part of the 1Science batch
- Affiliation Label Tesim:
- Department of Epidemiology
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/4rpk-rb22
- Identifier:
- https//dx.doi.org/10.1074/jbc.M901547200
- ISSN:
- 0021-9258
- Journal Issue:
- 26
- Journal Title:
- The Journal of Biological Chemistry
- Journal Volume:
- 284
- Keyword:
- Nucleocapsid Proteins, Peptide Library, Humans, Fluorescent Antibody Technique, RNA, Messenger, Antibodies, Monoclonal, Gene Expression Profiling, Amino Acid Sequence, Cells, Cultured, Cercopithecus aethiops, Molecular Sequence Data, Kidney, Fibronectins, Animals, and Immunoprecipitation
- Language Label:
- English
- ORCID:
- Other Affiliation:
- University of California at Los Angeles, University of Southern California, California Institute of Technology, and
- Page End:
- 17520
- Page Start:
- 17512
- Person:
- Deng, Hongyu, Sun, Ren, Liao, Hsiang-I, Roberts, Richard W., Olson, C. Anders, Wong, Elaine, Hwang, Seungmin, and Baric, Ralph S.
- Rights Statement Label:
- In Copyright
- Source:
- hq37vt14v
987. Molecular pathology of emerging coronavirus infections
- Title Tesim:
- Molecular pathology of emerging coronavirus infections
- Creator:
- Lisa E Gralinski and Ralph S Baric
- Date of publication:
- 2014
- Abstract Tesim:
- Respiratory viruses can cause a wide spectrum of pulmonary diseases, ranging from mild, upper respiratory tract infections to severe and life-threatening lower respiratory tract infections, including the development of acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). Viral clearance and subsequent recovery from infection require activation of an effective host immune response; however, many immune effector cells may also cause injury to host tissues. Severe acute respiratory syndrome (SARS) coronavirus and Middle East respiratory syndrome (MERS) coronavirus cause severe infection of the lower respiratory tract, with 10% and 35% overall mortality rates, respectively; however, >50% mortality rates are seen in the aged and immunosuppressed populations. While these viruses are susceptible to interferon treatment in vitro, they both encode numerous genes that allow for successful evasion of the host immune system until after high virus titres have been achieved. In this review, we discuss the importance of the innate immune response and the development of lung pathology following human coronavirus infection.
- Resource type:
- Article
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/dnj5-r081
- ISSN:
- 1096-9896 and 0022-3417
- Journal Issue:
- 2
- Journal Title:
- Journal of Pathology
- Journal Volume:
- 235
- Keyword:
- acute lung injury, Host-Pathogen Interactions, Coronavirus Infections, Respiratory System, Risk factors, type II pneumocytes , Communicable Diseases, SARS-CoV, Virulence, Virology, Immunity, MERS-CoV, coronavirus, Antiviral Agents, acute respiratory distress syndrome, Biopsy, SARS Virus, Emerging, Humans, Severe Acute Respiratory Syndrome, Invited Reviews, Animals, ARDS, Predictive Value of Tests, and Innate
- Language Label:
- English
- ORCID:
- Other Affiliation:
- Department of Epidemiology, University of North Carolina, Chapel Hill, NC, USA and Department of Epidemiology, University of North Carolina, Chapel Hill, NC, USA ; Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, NC, USA
- Person:
- Lisa E Gralinski and Ralph S Baric
- Publisher:
- Wiley Online Library
- Rights Statement Label:
- In Copyright
- Source:
- fj236671d
988. Severe Acute Respiratory Syndrome Coronavirus Papain-Like Protease Ubiquitin-Like Domain and Catalytic Domain Regulate Antagonism of IRF3 and NF- B Signaling
- Title Tesim:
- Severe Acute Respiratory Syndrome Coronavirus Papain-Like Protease Ubiquitin-Like Domain and Catalytic Domain Regulate Antagonism of IRF3 and NF- B Signaling
- Creator:
- Frieman, Matthew, Ratia, Kiira, Johnston, Robert E., Mesecar, Andrew D., and Baric, Ralph S.
- Date of publication:
- 2009
- Abstract Tesim:
- The outcome of a viral infection is regulated in part by the complex coordination of viral and host interactions that compete for the control and optimization of virus replication. Severe acute respiratory syndrome coronavirus (SARS-CoV) intimately engages and regulates the host innate immune responses during infection. Using a novel interferon (IFN) antagonism screen, we show that the SARS-CoV proteome contains several replicase, structural, and accessory proteins that antagonize the IFN pathway. In this study, we focus on the SARS-CoV papain-like protease (PLP), which engages and antagonizes the IFN induction and NF-κB signaling pathways. PLP blocks these pathways by affecting activation of the important signaling proteins in each pathway, IRF3 and NF-κB. We also show that the ubiquitin-like domain of PLP is necessary for pathway antagonism but not sufficient by itself to block these pathways regardless of the enzymatic activity of the protease. The potential mechanism of PLP antagonism and its role in pathogenesis are discussed.
- Resource type:
- Article
- Admin Note:
- deposited as part of the 1Science batch
- Affiliation Label Tesim:
- Department of Epidemiology and Carolina Vaccine Institute
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/7gc9-pv50
- Identifier:
- https://dx.doi.org/10.1128/JVI.02220-08
- ISSN:
- 0022-538X
- Journal Issue:
- 13
- Journal Title:
- Journal of Virology
- Journal Volume:
- 83
- Keyword:
- Regulation (control), Ubiquitin, Peptidases, Respiratory disease, Hydrolases, Transcription factor NFkB, Lung disease, Enzyme, Viral disease, Virology, Cysteine endopeptidases, Severe acute respiratory syndrome, Coronavirus, Infection, Papain, and virus
- Language Label:
- English
- ORCID:
- Other Affiliation:
- and University of Illinois, Chicago
- Page End:
- 6705
- Page Start:
- 6689
- Person:
- Frieman, Matthew, Ratia, Kiira, Johnston, Robert E., Mesecar, Andrew D., and Baric, Ralph S.
- Rights Statement Label:
- In Copyright
- Source:
- 9593v103b
989. Prophylactic and postexposure efficacy of a potent human monoclonal antibody against MERS coronavirus
- Title Tesim:
- Prophylactic and postexposure efficacy of a potent human monoclonal antibody against MERS coronavirus
- Creator:
- Jincun Zhao, Antonio Lanzavecchia, Davide Corti, Maria Zambon, Mattia Pedotti, Fabrizia Vanzetta, Nicholas A Barrett, Stanley Perlman, Luca Simonelli, Craig Fett, Luca Varani, Christopher J. Langrish, Blanca Fernandez-Rodriguez, Mathilde Foglierini, Sudhakar Agnihothram, Ralph S. Baric, Gloria Agatic, Federica Sallusto, and Robin Gopal
- Date of publication:
- 2015
- Abstract Tesim:
- Middle East Respiratory Syndrome coronavirus (MERS-CoV) causes severe respiratory disease with a high mortality rate. There is no licensed vaccine or antiviral for MERS. Here we isolated for the first time, to our knowledge, a potent MERS-CoV–neutralizing antibody from memory B cells of an infected individual. This antibody binds to a novel site on the viral Spike protein, neutralizes by interfering with the binding to the cellular receptor CD26, and is highly effective both in prophylaxis and in therapy in a relevant mouse model. This antibody can be developed for prophylaxis, for postexposure prophylaxis, or for the treatment of severe MERS-CoV infections.
- Resource type:
- Article
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/m5rz-av17
- ISSN:
- 1091-6490, 0027-8424, and 0744-2831
- Journal Issue:
- 33
- Journal Title:
- Proceedings of the National Academy of Sciences of the United States of America
- Journal Volume:
- 112
- Keyword:
- Animals, Cricetinae, Dipeptidyl Peptidase 4, Female, Cricetulus, Coronavirus Infections, Humans, Antibodies, Viral, Antibodies, Monoclonal, Antibodies, Neutralizing, emerging viruses, CHO Cells, Enzyme-Linked Immunosorbent Assay, B-Lymphocytes, neutralizing antibody, serotherapy, Binding Sites, Amino Acid Sequence, and MERS-CoV
- Language Label:
- English
- ORCID:
- Other Affiliation:
- Department of Microbiology, University of Iowa, Iowa City, IA 52240 ; State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510120, China, Immune Regulation Unit, Institute for Research in Biomedicine, Universitàdella Svizzera Italiana, 6500 Bellinzona, Switzerland ; Institute of Microbiology, Eidgenössische Technische Hochschule Zurich, 8093 Zurich, Switzerland, Immune Regulation Unit, Institute for Research in Biomedicine, Universitàdella Svizzera Italiana, 6500 Bellinzona, Switzerland ; Humabs BioMed SA, 6500 Bellinzona, Switzerland, Microbiology Services Colindale, Public Health England, London NW9 5HT, United Kingdom, Immune Regulation Unit, Institute for Research in Biomedicine, Universitàdella Svizzera Italiana, 6500 Bellinzona, Switzerland, Humabs BioMed SA, 6500 Bellinzona, Switzerland, Guy’s and St. Thomas’ National Health Service Foundation Trust, London SE1 7EH, United Kingdom; King’s College London, Strand, London WC2R 2LS, United Kingdom, Department of Microbiology, University of Iowa, Iowa City, IA 52240, Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, Guy’s and St. Thomas’ National Health Service Foundation Trust, London SE1 7EH, United Kingdom, and Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599 ; Department of Epidemiology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599
- Page End:
- 10478
- Page Start:
- 10473
- Person:
- Jincun Zhao, Antonio Lanzavecchia, Davide Corti, Maria Zambon, Mattia Pedotti, Fabrizia Vanzetta, Nicholas A Barrett, Stanley Perlman, Luca Simonelli, Craig Fett, Luca Varani, Christopher J. Langrish, Blanca Fernandez-Rodriguez, Mathilde Foglierini, Sudhakar Agnihothram, Ralph S. Baric, Gloria Agatic, Federica Sallusto, and Robin Gopal
- Publisher:
- Proceedings of the National Academy of Sciences of the United States of America
- Rights Statement Label:
- In Copyright
- Source:
- xs55mh661
- Subject:
- Coronavirus infections and Coronaviruses
990. Severe Acute Respiratory Syndrome Coronavirus nsp9 Dimerization Is Essential for Efficient Viral Growth
- Title Tesim:
- Severe Acute Respiratory Syndrome Coronavirus nsp9 Dimerization Is Essential for Efficient Viral Growth
- Creator:
- Schultz, L. Wayne, Umland, Timothy C., Donaldson, Eric F., Rimmer, Ryan A., Baric, Ralph S., and Mikinis, Zachary J.
- Date of publication:
- 2009
- Abstract Tesim:
- The severe acute respiratory syndrome coronavirus (SARS-CoV) devotes a significant portion of its genome to producing nonstructural proteins required for viral replication. SARS-CoV nonstructural protein 9 (nsp9) was identified as an essential protein with RNA/DNA-binding activity, and yet its biological function within the replication complex remains unknown. Nsp9 forms a dimer through the interaction of parallel α-helices containing the protein-protein interaction motif GXXXG. In order to study the role of the nsp9 dimer in viral reproduction, residues G100 and G104 at the helix interface were targeted for mutation. Multi-angle light scattering measurements indicated that G100E, G104E, and G104V mutants are monomeric in solution, thereby disrupting the dimer. However, electrophoretic mobility assays revealed that the mutants bound RNA with similar affinity. Further experiments using fluorescence anisotropy showed a 10-fold reduction in RNA binding in the G100E and G104E mutants, whereas the G104V mutant had only a 4-fold reduction. The structure of G104E nsp9 was determined to 2.6-Å resolution, revealing significant changes at the dimer interface. The nsp9 mutations were introduced into SARS-CoV using a reverse genetics approach, and the G100E and G104E mutations were found to be lethal to the virus. The G104V mutant produced highly debilitated virus and eventually reverted back to the wild-type protein sequence through a codon transversion. Together, these data indicate that dimerization of SARS-CoV nsp9 at the GXXXG motif is not critical for RNA binding but is necessary for viral replication.
- Resource type:
- Article
- Admin Note:
- deposited as part of 1Science batch
- Affiliation Label Tesim:
- Department of Microbiology and Immunology
- Type:
- http://purl.org/dc/dcmitype/Text
- DOI:
- https://doi.org/10.17615/map2-d042
- Identifier:
- https://dx.doi.org/10.1128/JVI.01505-08
- ISSN:
- 0022-538X
- Journal Issue:
- 7
- Journal Title:
- Journal of Virology
- Journal Volume:
- 83
- Keyword:
- Virus, Lung disease, Virology, Respiratory disease, Severe acute respiratory syndrome, Coronavirus, Viral disease, Infection, and Dimerization
- Language Label:
- English
- ORCID:
- Other Affiliation:
- State University of New York at Buffalo, , and Hauptman-Woodward Medical Research Institute
- Page End:
- 3018
- Page Start:
- 3007
- Person:
- Schultz, L. Wayne, Umland, Timothy C., Donaldson, Eric F., Rimmer, Ryan A., Baric, Ralph S., and Mikinis, Zachary J.
- Rights Statement Label:
- In Copyright
- Source:
- 4b29bb291
Collection Details
- Total items
-
1183
- Size
-
unknown
- Keyword
- SARS-CoV
severe acute respiratory syndrome
MERS-CoV
middle east respiratory syndrome
coronavirus
covid-19
SARS-CoV-2
- Date created
-
April 16, 2020
- Language
- English
- Identifier
- https://doi.org/10.17615/krdy-6769