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Article: Crucial mutation in the exoribonuclease domain of nsp14 of PEDV leads to high genetic instability during viral replication.

Niu, Xiaoyu / Kong, Fanzhi / Hou, Yixuan J / Wang, Qiuhong

2021 Volume 11, Issue 1, Page(s) 106

Abstract: Background: Coronavirus (CoV) nonstructural protein 14 (nsp14) has exoribonuclease (ExoN) activity, responsible for proofreading and contributing to replication fidelity. It has been reported that CoVs exhibit variable sensitivity to nsp14-ExoN ... ...

Abstract	Background: Coronavirus (CoV) nonstructural protein 14 (nsp14) has exoribonuclease (ExoN) activity, responsible for proofreading and contributing to replication fidelity. It has been reported that CoVs exhibit variable sensitivity to nsp14-ExoN deficiency. Betacoronavirus murine hepatitis virus (MHV) and severe acute respiratory syndrome (SARS)-CoV were viable upon nsp14-ExoN deficiency. While betacoronavirus Middle East respiratory syndrome (MERS)-CoV and SARS-CoV-2 were non-viable with disabled nsp14-ExoN. In this study, we investigated the nsp14-ExoN deficiency of alphacoronavirus porcine epidemic diarrhea virus (PEDV) in viral pathogenesis using reverse genetics. Results: Eight nsp14-ExoN deficient mutants, targeting the predicted active sites and the Zinc finger or mental-coordinating sites, of PEDV were designed. Only one mutant E191A with a mutation in the Mg Conclusion: The recombinant PEDV variants carrying mutations at the essential functional sites within nsp14-ExoN were either lethal or genetically unstable. Our finding further confirmed the critical role of nsp14-ExoN in CoV life cycle, suggesting that it may be a target for the design of universal anti-CoV drugs.
Language	English
Publishing date	2021-06-07
Publishing country	England
Document type	Journal Article
ZDB-ID	2593367-X
ISSN	2045-3701
ISSN	2045-3701
DOI	10.1186/s13578-021-00598-1
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Article ; Online: A recombination-resistant genome for live attenuated and stable PEDV vaccines by engineering the transcriptional regulatory sequences.

Niu, Xiaoyu / Liu, Mingde / Yang, Shaomin / Xu, Jiayu / Hou, Yixuan J / Liu, Dongxiao / Tang, Qiyi / Zhu, Hua / Wang, Qiuhong

Journal of virology

2023 Volume 97, Issue 12, Page(s) e0119323

Abstract: Importance: Coronaviruses are important pathogens of humans and animals, and vaccine developments against them are imperative. Due to the ability to induce broad and prolonged protective immunity and the convenient administration routes, live attenuated ...

Abstract	Importance: Coronaviruses are important pathogens of humans and animals, and vaccine developments against them are imperative. Due to the ability to induce broad and prolonged protective immunity and the convenient administration routes, live attenuated vaccines (LAVs) are promising arms for controlling the deadly coronavirus infections. However, potential recombination events between vaccine and field strains raise a safety concern for LAVs. The porcine epidemic diarrhea virus (PEDV) remodeled TRS (RMT) mutant generated in this study replicated efficiently in both cell culture and in pigs and retained protective immunogenicity against PEDV challenge in pigs. Furthermore, the RMT PEDV was resistant to recombination and genetically stable. Therefore, RMT PEDV can be further optimized as a backbone for the development of safe LAVs.
MeSH term(s)	Animals ; Coronavirus Infections/immunology ; Coronavirus Infections/prevention & control ; Coronavirus Infections/veterinary ; Porcine epidemic diarrhea virus/genetics ; Porcine epidemic diarrhea virus/growth & development ; Porcine epidemic diarrhea virus/immunology ; Recombination, Genetic ; Swine/immunology ; Swine/virology ; Swine Diseases/immunology ; Swine Diseases/prevention & control ; Swine Diseases/virology ; Vaccines, Attenuated/genetics ; Vaccines, Attenuated/immunology ; Viral Vaccines/genetics ; Viral Vaccines/immunology ; Virus Replication ; Cells, Cultured ; Mutation
Chemical Substances	Vaccines, Attenuated ; Viral Vaccines
Language	English
Publishing date	2023-11-16
Publishing country	United States
Document type	Journal Article
ZDB-ID	80174-4
ISSN	1098-5514 ; 0022-538X
ISSN (online)	1098-5514
ISSN	0022-538X
DOI	10.1128/jvi.01193-23
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Article ; Online: Crucial mutation in the exoribonuclease domain of nsp14 of PEDV leads to high genetic instability during viral replication

Xiaoyu Niu / Fanzhi Kong / Yixuan J. Hou / Qiuhong Wang

Cell & Bioscience, Vol 11, Iss 1, Pp 1-

2021 Volume 13

Abstract: Abstract Background Coronavirus (CoV) nonstructural protein 14 (nsp14) has exoribonuclease (ExoN) activity, responsible for proofreading and contributing to replication fidelity. It has been reported that CoVs exhibit variable sensitivity to nsp14-ExoN ... ...

Abstract	Abstract Background Coronavirus (CoV) nonstructural protein 14 (nsp14) has exoribonuclease (ExoN) activity, responsible for proofreading and contributing to replication fidelity. It has been reported that CoVs exhibit variable sensitivity to nsp14-ExoN deficiency. Betacoronavirus murine hepatitis virus (MHV) and severe acute respiratory syndrome (SARS)-CoV were viable upon nsp14-ExoN deficiency. While betacoronavirus Middle East respiratory syndrome (MERS)-CoV and SARS-CoV-2 were non-viable with disabled nsp14-ExoN. In this study, we investigated the nsp14-ExoN deficiency of alphacoronavirus porcine epidemic diarrhea virus (PEDV) in viral pathogenesis using reverse genetics. Results Eight nsp14-ExoN deficient mutants, targeting the predicted active sites and the Zinc finger or mental-coordinating sites, of PEDV were designed. Only one mutant E191A with a mutation in the Mg2+-binding site was rescued using the infectious clone of PEDV PC22A strain (icPC22A). The passage no.1–3 (P1-3) of E191A grew to very low titers in Vero cells. To evaluate the pathogenesis of the E191A, 4 or 5-day-old gnotobiotic pigs were inoculated orally with 100 TCID50/pig of the E191A-P1, icPC22A, or mock. All mock pigs did not shed virus in feces or show clinical signs. All pigs inoculated with icPC22A shed high viral RNA levels, had severe diarrhea, and died by 6 days post-inoculation (dpi). In contrast, only 3 pigs (3/4, 75%) in the E191A-P1 group shed low levels of viral RNA and 2 pigs had moderate diarrhea at acute infection phase. At 22 dpi, each pig was challenged orally with 106 plaque forming unit of virulent icPC22A. All pigs in the mock group developed severe diarrhea and 2 of the 5 pigs died. Pigs in the E191A-P1 group had less severe diarrhea and no pigs died. Sanger sequencing analysis revealed that the viral genome in the fecal sample of one E191A-P1-inoculated pig and the P4 virus passaged in vitro lost the E191A mutation, suggesting the genetic instability of the E191A mutant. Conclusion The recombinant PEDV variants ...
Keywords	Porcine epidemic diarrhea virus ; Coronavirus ; Nsp14 ; Exoribonuclease ; Reverse genetics ; Infectious clone ; Biotechnology ; TP248.13-248.65 ; Biology (General) ; QH301-705.5 ; Biochemistry ; QD415-436
Subject code	630
Language	English
Publishing date	2021-06-01T00:00:00Z
Publisher	BMC
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Chimeric Porcine Deltacoronaviruses with Sparrow Coronavirus Spike Protein or the Receptor-Binding Domain Infect Pigs but Lose Virulence and Intestinal Tropism.

Niu, Xiaoyu / Hou, Yixuan J / Jung, Kwonil / Kong, Fanzhi / Saif, Linda J / Wang, Qiuhong

Viruses

2021 Volume 13, Issue 1

Abstract: Porcine deltacoronavirus (PDCoV) strain OH-FD22 infects poultry and shares high nucleotide identity with sparrow-origin deltacoronaviruses (SpDCoV) ISU73347 and HKU17 strains. We hypothesized that the spike (S) protein or receptor-binding domain (RBD) ... ...

Abstract	Porcine deltacoronavirus (PDCoV) strain OH-FD22 infects poultry and shares high nucleotide identity with sparrow-origin deltacoronaviruses (SpDCoV) ISU73347 and HKU17 strains. We hypothesized that the spike (S) protein or receptor-binding domain (RBD) from these SpDCoVs would alter the host and tissue tropism of PDCoV. First, an infectious cDNA clone of PDCoV OH-FD22 strain (icPDCoV) was generated and used to construct chimeric icPDCoVs harboring the S protein of HKU17 (icPDCoV-S
MeSH term(s)	Amino Acid Motifs ; Animals ; Bird Diseases/virology ; Cell Line ; Coronavirus Infections/pathology ; Deltacoronavirus/genetics ; Deltacoronavirus/pathogenicity ; Intestines/pathology ; Intestines/virology ; Recombinant Proteins/genetics ; Respiratory System/pathology ; Respiratory System/virology ; Sparrows ; Spike Glycoprotein, Coronavirus/genetics ; Swine ; Swine Diseases/virology ; Viral Tropism/genetics ; Virulence/genetics
Chemical Substances	Recombinant Proteins ; Spike Glycoprotein, Coronavirus
Language	English
Publishing date	2021-01-17
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v13010122
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Article: A semi-quantitative, rapid, point of care SARS-CoV-2 serologic assay predicts neutralizing antibody levels.

Markmann, Alena J / Bhowmik, D Ryan / Jiang, Baowei / Van Hoy, Michael / Wang, Frank / Hou, Yixuan J / Baric, Ralph S / de Silva, Aravinda M / Bartelt, Luther A

bioRxiv : the preprint server for biology

2023

Abstract: The ongoing COVID-19 pandemic has caused millions of deaths and the continued emergence of new variants suggests continued circulation in the human population. In the current time of vaccine availability and new therapeutic development, including ... ...

Abstract	The ongoing COVID-19 pandemic has caused millions of deaths and the continued emergence of new variants suggests continued circulation in the human population. In the current time of vaccine availability and new therapeutic development, including antibody-based therapies, many questions about long-term immunity and protection remain uncertain. Identification of protective antibodies in individuals is often done using highly specialized and challenging assays such as functional neutralizing assays, which are not available in the clinical setting. Therefore, there is a great need for the development of rapid, clinically available assays that correlate with neutralizing antibody assays to identify individuals who may benefit from additional vaccination or specific COVID-19 therapies. In this report, we apply a novel semi-quantitative method to an established lateral flow assay (sqLFA) and analyze its ability to detect the presence functional neutralizing antibodies from the serum of COVID-19 recovered individuals. We found that the sqLFA has a strong positive correlation with neutralizing antibody levels. At lower assay cutoffs, the sqLFA is a highly sensitive assay to identify the presence of a range of neutralizing antibody levels. At higher cutoffs, it can detect higher levels of neutralizing antibody with high specificity. This sqLFA can be used both as a screening tool to identify individuals with any level of neutralizing antibody to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), or as a more specific tool to identify those with high neutralizing antibody levels who may not benefit from antibody-based therapies or further vaccination.
Language	English
Publishing date	2023-06-01
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.05.30.542314
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Article ; Online: Critical ACE2 Determinants of SARS-CoV-2 and Group 2B Coronavirus Infection and Replication.

Adams, Lily E / Dinnon, Kenneth H / Hou, Yixuan J / Sheahan, Timothy P / Heise, Mark T / Baric, Ralph S

mBio

2021 Volume 12, Issue 2

Abstract: The angiotensin-converting enzyme 2 (ACE2) receptor is a major severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) host range determinant, and understanding SARS-CoV-2-ACE2 interactions will provide important insights into COVID-19 pathogenesis ... ...

Abstract	The angiotensin-converting enzyme 2 (ACE2) receptor is a major severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) host range determinant, and understanding SARS-CoV-2-ACE2 interactions will provide important insights into COVID-19 pathogenesis and animal model development. SARS-CoV-2 cannot infect mice due to incompatibility between its receptor binding domain and the murine ACE2 receptor. Through molecular modeling and empirical
MeSH term(s)	Amino Acid Sequence ; Angiotensin-Converting Enzyme 2/chemistry ; Angiotensin-Converting Enzyme 2/genetics ; Angiotensin-Converting Enzyme 2/metabolism ; Animals ; Betacoronavirus/metabolism ; Betacoronavirus/physiology ; Binding Sites ; COVID-19/virology ; Cell Line ; Coronavirus Infections/virology ; Host Specificity ; Humans ; Mice ; Models, Molecular ; Mutation ; Protein Binding ; Recombinant Proteins/chemistry ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism ; SARS-CoV-2/metabolism ; SARS-CoV-2/physiology ; Spike Glycoprotein, Coronavirus/chemistry ; Spike Glycoprotein, Coronavirus/metabolism ; Virus Replication
Chemical Substances	Recombinant Proteins ; Spike Glycoprotein, Coronavirus ; Angiotensin-Converting Enzyme 2 (EC 3.4.17.23)
Language	English
Publishing date	2021-03-16
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2557172-2
ISSN	2150-7511 ; 2161-2129
ISSN (online)	2150-7511
ISSN	2161-2129
DOI	10.1128/mBio.03149-20
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Article ; Online: Validation of the Chinese version of the diabetes health profile to predict the impact of mobile health education on quality of life in type 2 diabetes patients.

Lyu, Xiaokang / Zeng, Jinmei / Lin, Jingna / Song, Yixuan / Yang, Tingting / Hou, Wenjing

Frontiers in public health

2024 Volume 12, Page(s) 1330154

Abstract: Purpose: The Diabetes Health Profile (DHP18), initially created in the United Kingdom, currently lacks a Chinese version. This study endeavors to authenticate the Chinese adaptation of the DHP18 and assess the influence of mobile health (mHealth) ... ...

Abstract	Purpose: The Diabetes Health Profile (DHP18), initially created in the United Kingdom, currently lacks a Chinese version. This study endeavors to authenticate the Chinese adaptation of the DHP18 and assess the influence of mobile health (mHealth) education intervention on the quality of life of individuals living with diabetes. Patients and methods: The study included 470 Type 2 diabetes Mellitus (T2DM) patients (204 men, 266 women), spanning an age range of 19-79 years, with an average age of 54 ± 12.40 years. Data analysis employed Jamovie and Mplus software. Moreover, test-retest reliability was evaluated in 52 hospitalized T2DM patients through two repeated measurements taken 4 weeks apart. Results: The Chinese version DHP18 scale exhibited high reliability, evidenced by a Cronbach's alpha of 0.88, and coefficient of test-retest reliability of 0.84. Individual subscales also demonstrated strong reliability, ranging from 0.76 to 0.84, with test-retest reliability spanning from 0.71 to 0.74. Confirmatory Factor Analysis (CFA) employing a three-factor structure (χ Conclusion: The Chinese version of the Diabetes Health Profile Scale meets stringent psychometric standards and stands as an appropriate measurement tool for Chinese T2DM patients, maintaining comparable results to the original scale's structure. The mHealth education intervention yielded a notably positive impact on the quality of life among T2DM patients. Mediation analysis revealed that the three dimensions of the DHP were mediated by Appraisal of Diabetes and Diabetes Self-Management Efficacy, partially mediated by Psychological Distress and Behavior Adherence, and fully mediated by Dietary Abstinence, providing insight into the positive effects of the mHealth model on the quality of life of diabetic patients.
MeSH term(s)	Male ; Humans ; Female ; Adult ; Middle Aged ; Aged ; Young Adult ; Diabetes Mellitus, Type 2/therapy ; Quality of Life ; Reproducibility of Results ; Health Education ; Educational Status
Language	English
Publishing date	2024-02-21
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2711781-9
ISSN	2296-2565 ; 2296-2565
ISSN (online)	2296-2565
ISSN	2296-2565
DOI	10.3389/fpubh.2024.1330154
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Article ; Online: Public Attitudes and Factors of COVID-19 Testing Hesitancy in the United Kingdom and China: Comparative Infodemiology Study.

Lin, Leesa / Song, Yi / Wang, Qian / Pu, Jialu / Sun, Fiona Yueqian / Zhang, Yixuan / Zhou, Xinyu / Larson, Heidi J / Hou, Zhiyuan

JMIR infodemiology

2021 Volume 1, Issue 1, Page(s) e26895

Abstract: Background: Massive community-wide testing has become the cornerstone of management strategies for the COVID-19 pandemic.: Objective: This study was a comparative analysis between the United Kingdom and China, which aimed to assess public attitudes ... ...

Abstract	Background: Massive community-wide testing has become the cornerstone of management strategies for the COVID-19 pandemic. Objective: This study was a comparative analysis between the United Kingdom and China, which aimed to assess public attitudes and uptake regarding COVID-19 testing, with a focus on factors of COVID-19 testing hesitancy, including effectiveness, access, risk perception, and communication. Methods: We collected and manually coded 3856 UK tweets and 9299 Chinese Sina Weibo posts mentioning COVID-19 testing from June 1 to July 15, 2020. Adapted from the World Health Organization's 3C Model of Vaccine Hesitancy, we employed social listening analysis examining key factors of COVID-19 testing hesitancy (confidence, complacency, convenience, and communication). Descriptive analysis, time trends, geographical mapping, and chi-squared tests were performed to assess the temporal, spatial, and sociodemographic characteristics that determine the difference in attitudes or uptake of COVID-19 tests. Results: The UK tweets demonstrated a higher percentage of support toward COVID-19 testing than the posts from China. There were much wider reports of public uptake of COVID-19 tests in mainland China than in the United Kingdom; however, uncomfortable experiences and logistical barriers to testing were more expressed in China. The driving forces for undergoing COVID-19 testing were personal health needs, community-wide testing, and mandatory testing policies for travel, with major differences in the ranking order between the two countries. Rumors and information inquiries about COVID-19 testing were also identified. Conclusions: Public attitudes and acceptance toward COVID-19 testing constantly evolve with local epidemic situations. Policies and information campaigns that emphasize the importance of timely testing and rapid communication responses to inquiries and rumors, and provide a supportive environment for accessing tests are key to tackling COVID-19 testing hesitancy and increasing uptake.
Language	English
Publishing date	2021-08-27
Publishing country	Canada
Document type	Journal Article
ISSN	2564-1891
ISSN (online)	2564-1891
DOI	10.2196/26895
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Article ; Online: A MERS-CoV antibody neutralizes a pre-emerging group 2c bat coronavirus.

Tse, Longping V / Hou, Yixuan J / McFadden, Elizabeth / Lee, Rhianna E / Scobey, Trevor D / Leist, Sarah R / Martinez, David R / Meganck, Rita M / Schäfer, Alexandra / Yount, Boyd L / Mascenik, Teresa / Powers, John M / Randell, Scott H / Zhang, Yi / Wang, Lingshu / Mascola, John / McLellan, Jason S / Baric, Ralph S

Science translational medicine

2023 Volume 15, Issue 715, Page(s) eadg5567

Abstract: The repeated emergence of zoonotic human betacoronaviruses (β-CoVs) dictates the need for broad therapeutics and conserved epitope targets for countermeasure design. Middle East respiratory syndrome (MERS)-related coronaviruses (CoVs) remain a pressing ... ...

Abstract	The repeated emergence of zoonotic human betacoronaviruses (β-CoVs) dictates the need for broad therapeutics and conserved epitope targets for countermeasure design. Middle East respiratory syndrome (MERS)-related coronaviruses (CoVs) remain a pressing concern for global health preparedness. Using metagenomic sequence data and CoV reverse genetics, we recovered a full-length wild-type MERS-like BtCoV/
MeSH term(s)	Humans ; Animals ; Mice ; Middle East Respiratory Syndrome Coronavirus ; Dipeptidyl Peptidase 4/genetics ; Dipeptidyl Peptidase 4/metabolism ; Chiroptera ; Cryoelectron Microscopy ; Coronavirus Infections ; Antibodies, Monoclonal/metabolism
Chemical Substances	Dipeptidyl Peptidase 4 (EC 3.4.14.5) ; Antibodies, Monoclonal
Language	English
Publishing date	2023-09-27
Publishing country	United States
Document type	Journal Article
ZDB-ID	2518854-9
ISSN	1946-6242 ; 1946-6234
ISSN (online)	1946-6242
ISSN	1946-6234
DOI	10.1126/scitranslmed.adg5567
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Article ; Online: Multivalent S2-based vaccines provide broad protection against SARS-CoV-2 variants of concern and pangolin coronaviruses.

Halfmann, Peter J / Frey, Steven J / Loeffler, Kathryn / Kuroda, Makoto / Maemura, Tadashi / Armbrust, Tammy / Yang, Jie E / Hou, Yixuan J / Baric, Ralph / Wright, Elizabeth R / Kawaoka, Yoshihiro / Kane, Ravi S

EBioMedicine

2022 Volume 86, Page(s) 104341

Abstract: Background: The COVID-19 pandemic continues to cause morbidity and mortality worldwide. Most approved COVID-19 vaccines generate a neutralizing antibody response that primarily targets the highly variable receptor-binding domain (RBD) of the SARS-CoV-2 ... ...

Abstract	Background: The COVID-19 pandemic continues to cause morbidity and mortality worldwide. Most approved COVID-19 vaccines generate a neutralizing antibody response that primarily targets the highly variable receptor-binding domain (RBD) of the SARS-CoV-2 spike (S) protein. SARS-CoV-2 "variants of concern" have acquired mutations in this domain allowing them to evade vaccine-induced humoral immunity. Recent approaches to improve the breadth of protection beyond SARS-CoV-2 have required the use of mixtures of RBD antigens from different sarbecoviruses. It may therefore be beneficial to develop a vaccine in which the protective immune response targets a more conserved region of the S protein. Methods: Here we have developed a vaccine based on the conserved S2 subunit of the S protein and optimized the adjuvant and immunization regimen in Syrian hamsters and BALB/c mice. We have characterized the efficacy of the vaccine against SARS-CoV-2 variants and other coronaviruses. Findings: Immunization with S2-based constructs elicited a broadly cross-reactive IgG antibody response that recognized the spike proteins of not only SARS-CoV-2 variants, but also SARS-CoV-1, and the four endemic human coronaviruses. Importantly, immunization reduced virus titers in respiratory tissues in vaccinated animals challenged with SARS-CoV-2 variants B.1.351 (beta), B.1.617.2 (delta), and BA.1 (omicron) as well as a pangolin coronavirus. Interpretation: These results suggest that S2-based constructs can elicit a broadly cross-reactive antibody response resulting in limited virus replication, thus providing a framework for designing vaccines that elicit broad protection against coronaviruses. Funding: NIH, Japan Agency for Medical Research and Development, Garry Betty/ V Foundation Chair Fund, and NSF.
MeSH term(s)	Animals ; Cricetinae ; Mice ; Humans ; SARS-CoV-2/genetics ; Vaccines, Combined ; COVID-19 Vaccines ; Pangolins ; Pandemics ; COVID-19/prevention & control ; Antibodies, Neutralizing ; Antibodies, Viral
Chemical Substances	Vaccines, Combined ; COVID-19 Vaccines ; Antibodies, Neutralizing ; Antibodies, Viral
Language	English
Publishing date	2022-11-11
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2851331-9
ISSN	2352-3964
ISSN (online)	2352-3964
DOI	10.1016/j.ebiom.2022.104341
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