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      Persistence of serum and saliva antibody responses to SARS-CoV-2 spike antigens in COVID-19 patients

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      1 , 2 , 3 , 1 , 4 , 5 , 2 , 2 , 3 , 1 , 1 , 1 , 1 , 6 , 7 , 2 , 2 , 3 , 6 , 8 , 5 , 5 , 1 , 9 , 2 , 2 , 3 , 2 , 2 , 3 , 10 , 3 , 6 , 11 , 9 , 2 , 1 , 12 , 13 , 3 , 14 , 7 , 2 , 4 , 5 , 1 , * , 2 , 3 , *
      Science Immunology
      American Association for the Advancement of Science

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          Abstract

          Saliva is an alternative biofluid to serum for detecting and monitoring IgG to SARS-CoV-2 spike and RBD antigens in COVID-19.

          Abstract

          While the antibody response to SARS-CoV-2 has been extensively studied in blood, relatively little is known about the antibody response in saliva and its relationship to systemic antibody levels. Here, we profiled by enzyme-linked immunosorbent assays (ELISAs) IgG, IgA and IgM responses to the SARS-CoV-2 spike protein (full length trimer) and its receptor-binding domain (RBD) in serum and saliva of acute and convalescent patients with laboratory-diagnosed COVID-19 ranging from 3–115 days post-symptom onset (PSO), compared to negative controls. Anti-SARS-CoV-2 antibody responses were readily detected in serum and saliva, with peak IgG levels attained by 16–30 days PSO. Longitudinal analysis revealed that anti-SARS-CoV-2 IgA and IgM antibodies rapidly decayed, while IgG antibodies remained relatively stable up to 105 days PSO in both biofluids. Lastly, IgG, IgM and to a lesser extent IgA responses to spike and RBD in the serum positively correlated with matched saliva samples. This study confirms that serum and saliva IgG antibodies to SARS-CoV-2 are maintained in the majority of COVID-19 patients for at least 3 months PSO. IgG responses in saliva may serve as a surrogate measure of systemic immunity to SARS-CoV-2 based on their correlation with serum IgG responses.

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          Most cited references39

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          Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation

          Structure of the nCoV trimeric spike The World Health Organization has declared the outbreak of a novel coronavirus (2019-nCoV) to be a public health emergency of international concern. The virus binds to host cells through its trimeric spike glycoprotein, making this protein a key target for potential therapies and diagnostics. Wrapp et al. determined a 3.5-angstrom-resolution structure of the 2019-nCoV trimeric spike protein by cryo–electron microscopy. Using biophysical assays, the authors show that this protein binds at least 10 times more tightly than the corresponding spike protein of severe acute respiratory syndrome (SARS)–CoV to their common host cell receptor. They also tested three antibodies known to bind to the SARS-CoV spike protein but did not detect binding to the 2019-nCoV spike protein. These studies provide valuable information to guide the development of medical counter-measures for 2019-nCoV. Science, this issue p. 1260
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            Virological assessment of hospitalized patients with COVID-2019

            Coronavirus disease 2019 (COVID-19) is an acute infection of the respiratory tract that emerged in late 20191,2. Initial outbreaks in China involved 13.8% of cases with severe courses, and 6.1% of cases with critical courses3. This severe presentation may result from the virus using a virus receptor that is expressed predominantly in the lung2,4; the same receptor tropism is thought to have determined the pathogenicity-but also aided in the control-of severe acute respiratory syndrome (SARS) in 20035. However, there are reports of cases of COVID-19 in which the patient shows mild upper respiratory tract symptoms, which suggests the potential for pre- or oligosymptomatic transmission6-8. There is an urgent need for information on virus replication, immunity and infectivity in specific sites of the body. Here we report a detailed virological analysis of nine cases of COVID-19 that provides proof of active virus replication in tissues of the upper respiratory tract. Pharyngeal virus shedding was very high during the first week of symptoms, with a peak at 7.11 × 108 RNA copies per throat swab on day 4. Infectious virus was readily isolated from samples derived from the throat or lung, but not from stool samples-in spite of high concentrations of virus RNA. Blood and urine samples never yielded virus. Active replication in the throat was confirmed by the presence of viral replicative RNA intermediates in the throat samples. We consistently detected sequence-distinct virus populations in throat and lung samples from one patient, proving independent replication. The shedding of viral RNA from sputum outlasted the end of symptoms. Seroconversion occurred after 7 days in 50% of patients (and by day 14 in all patients), but was not followed by a rapid decline in viral load. COVID-19 can present as a mild illness of the upper respiratory tract. The confirmation of active virus replication in the upper respiratory tract has implications for the containment of COVID-19.
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              Functional assessment of cell entry and receptor usage for SARS-CoV-2 and other lineage B betacoronaviruses

              Over the past 20 years, several coronaviruses have crossed the species barrier into humans, causing outbreaks of severe, and often fatal, respiratory illness. Since SARS-CoV was first identified in animal markets, global viromics projects have discovered thousands of coronavirus sequences in diverse animals and geographic regions. Unfortunately, there are few tools available to functionally test these viruses for their ability to infect humans, which has severely hampered efforts to predict the next zoonotic viral outbreak. Here, we developed an approach to rapidly screen lineage B betacoronaviruses, such as SARS-CoV and the recent SARS-CoV-2, for receptor usage and their ability to infect cell types from different species. We show that host protease processing during viral entry is a significant barrier for several lineage B viruses and that bypassing this barrier allows several lineage B viruses to enter human cells through an unknown receptor. We also demonstrate how different lineage B viruses can recombine to gain entry into human cells, and confirm that human ACE2 is the receptor for the recently emerging SARS-CoV-2.
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                Author and article information

                Contributors
                Role: Resources
                Journal
                Sci Immunol
                Sci Immunol
                SciImmunol
                immunology
                Science Immunology
                American Association for the Advancement of Science
                2470-9468
                8 October 2020
                08 October 2020
                : 5
                : 52
                : eabe5511
                Affiliations
                [1 ]Department of Immunology, University of Toronto, Toronto, ON, Canada.
                [2 ]Lunenfeld-Tanenbaum Research Institute at Mount Sinai Hospital, Sinai Health System, Toronto, ON, Canada.
                [3 ]Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.
                [4 ]Institute of Health Policy, Management and Evaluation, University of Toronto, Toronto, ON, Canada.
                [5 ]Department of Microbiology, at Mount Sinai Hospital, Sinai Health System, Toronto, ON, Canada.
                [6 ]Combined Containment Level 3 Unit, University of Toronto, Toronto, ON, Canada.
                [7 ]Mammalian Cell Expression, Human Health Therapeutics Research Centre, National Research Council Canada, Montréal, QC, Canada.
                [8 ]Institute of Medical Science, University of Toronto, Toronto, ON, Canada.
                [9 ]College of Dentistry, University of Saskatchewan, Saskatoon, SK, Canada.
                [10 ]Department of Laboratory Medicine and Molecular Diagnostics, Division of Microbiology, Sunnybrook Health Sciences Centre; Biological Sciences, Sunnybrook Research Institute; and Division of Infectious Diseases, Sunnybrook Health Sciences Centre, Toronto, ON, Canada; Department of Laboratory Medicine and Pathology, University of Toronto, Toronto, ON, Canada.
                [11 ]Canadian Blood Services, Edmonton, AB & Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB, Canada.
                [12 ]St. Michael’s Hospital, Toronto, ON, Canada; Li Ka Shing Knowledge Institute.
                [13 ]Department of Medicine, University of Toronto, Toronto, ON, Canada.
                [14 ]Department of Biochemistry, University of Toronto, Toronto, ON, Canada.
                Author notes
                [†]

                These authors contributed equally

                Author information
                https://orcid.org/0000-0003-0921-9052
                https://orcid.org/0000-0002-6704-453X
                https://orcid.org/0000-0001-7082-3027
                https://orcid.org/0000-0001-6453-4552
                https://orcid.org/0000-0001-9283-6072
                https://orcid.org/0000-0002-5054-4685
                https://orcid.org/0000-0003-2300-1658
                https://orcid.org/0000-0001-6319-1551
                https://orcid.org/0000-0002-6809-001X
                https://orcid.org/0000-0002-2674-9234
                https://orcid.org/0000-0002-8357-4717
                https://orcid.org/0000-0003-2108-4656
                https://orcid.org/0000-0001-9991-0757
                https://orcid.org/0000-0003-0932-0644
                https://orcid.org/0000-0002-8979-1659
                https://orcid.org/0000-0002-9824-2117
                https://orcid.org/0000-0001-5012-2311
                https://orcid.org/0000-0002-1477-3616
                https://orcid.org/0000-0003-2519-1109
                https://orcid.org/0000-0002-7369-1936
                https://orcid.org/0000-0002-0952-2409
                https://orcid.org/0000-0002-2268-4111
                https://orcid.org/0000-0003-4576-6168
                https://orcid.org/0000-0002-6090-4437
                Article
                abe5511
                10.1126/sciimmunol.abe5511
                8050884
                33033173
                22b9d6a8-2c7d-4527-8f26-52e6b6e10cb4
                Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY).

                This is an open-access article distributed under the terms of the Creative Commons Attribution license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 28 August 2020
                : 05 October 2020
                Funding
                Funded by: doi http://dx.doi.org/10.13039/100008762, Genome Canada;
                Award ID: Ontario Genomics OGI-139
                Funded by: doi http://dx.doi.org/10.13039/100013873, Government of Ontario;
                Award ID: Ontario Together grant
                Funded by: doi http://dx.doi.org/10.13039/501100000024, Canadian Institutes of Health Research;
                Award ID: #VR1-172711, #VR4–172732, FDN15992, FDN143301
                Funded by: doi http://dx.doi.org/10.13039/501100000196, Canada Foundation for Innovation;
                Award ID: N/A
                Funded by: Sinai Health System Foundation;
                Award ID: N/A
                Funded by: CIHR - COVID-19 Rapid Research Funding Opportunity - Diagnostics;
                Funded by: Sunnybrook Foundation;
                Award ID: Questcap
                Categories
                Research Articles
                Research Articles
                R-Articles
                Coronavirus

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