{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T04:59:10Z","timestamp":1779339550098,"version":"3.51.4"},"update-to":[{"DOI":"10.1371\/journal.ppat.1009064","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2021,5,3]],"date-time":"2021-05-03T00:00:00Z","timestamp":1620000000000}}],"reference-count":99,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2021,4,21]],"date-time":"2021-04-21T00:00:00Z","timestamp":1618963200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"DFG","award":["DFG Project-ID 369799452 - TRR237 A12"],"award-info":[{"award-number":["DFG Project-ID 369799452 - TRR237 A12"]}]},{"name":"DFG","award":["DFG Project-ID 118803580 - SFB 870 Z1"],"award-info":[{"award-number":["DFG Project-ID 118803580 - SFB 870 Z1"]}]},{"DOI":"10.13039\/501100005722","name":"LMU Munich","doi-asserted-by":"crossref","award":["F\u00f6FoLe M 09\/2017"],"award-info":[{"award-number":["F\u00f6FoLe M 09\/2017"]}],"id":[{"id":"10.13039\/501100005722","id-type":"DOI","asserted-by":"crossref"}]},{"name":"DFG","award":["DFG SFB 1021 project ID 197785619\/B12"],"award-info":[{"award-number":["DFG SFB 1021 project ID 197785619\/B12"]}]},{"name":"German Federal Ministry of Health"},{"DOI":"10.13039\/100009139","name":"Deutsches Zentrum f\u00fcr Infektionsforschung","doi-asserted-by":"publisher","award":["HZI2016Z9"],"award-info":[{"award-number":["HZI2016Z9"]}],"id":[{"id":"10.13039\/100009139","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plospathogens.org"],"crossmark-restriction":false},"short-container-title":["PLoS Pathog"],"abstract":"<jats:p>Vaccines of outstanding efficiency, safety, and public acceptance are needed to halt the current SARS-CoV-2 pandemic. Concerns include potential side effects caused by the antigen itself and safety of viral DNA and RNA delivery vectors. The large SARS-CoV-2 spike (S) protein is the main target of current COVID-19 vaccine candidates but can induce non-neutralizing antibodies, which might cause vaccination-induced complications or enhancement of COVID-19 disease. Besides, encoding of a functional S in replication-competent virus vector vaccines may result in the emergence of viruses with altered or expanded tropism. Here, we have developed a safe single round rhabdovirus replicon vaccine platform for enhanced presentation of the S receptor-binding domain (RBD). Structure-guided design was employed to build a chimeric minispike comprising the globular RBD linked to a transmembrane stem-anchor sequence derived from rabies virus (RABV) glycoprotein (G). Vesicular stomatitis virus (VSV) and RABV replicons encoding the minispike not only allowed expression of the antigen at the cell surface but also incorporation into the envelope of secreted non-infectious particles, thus combining classic vector-driven antigen expression and particulate virus-like particle (VLP) presentation. A single dose of a prototype replicon vaccine complemented with VSV G, VSV\u0394G-minispike-eGFP (G), stimulated high titers of SARS-CoV-2 neutralizing antibodies in mice, equivalent to those found in COVID-19 patients, and protected transgenic K18-hACE2 mice from COVID-19-like disease. Homologous boost immunization further enhanced virus neutralizing activity. The results demonstrate that non-spreading rhabdovirus RNA replicons expressing minispike proteins represent effective and safe alternatives to vaccination approaches using replication-competent viruses and\/or the entire S antigen.<\/jats:p>","DOI":"10.1371\/journal.ppat.1009064","type":"journal-article","created":{"date-parts":[[2021,4,21]],"date-time":"2021-04-21T22:47:21Z","timestamp":1619045241000},"page":"e1009064","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":30,"title":["Safe and effective two-in-one replicon-and-VLP minispike vaccine for COVID-19: Protection of mice after a single immunization"],"prefix":"10.1371","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3233-6145","authenticated-orcid":true,"given":"Alexandru A.","family":"Hennrich","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6908-1556","authenticated-orcid":true,"given":"Bevan","family":"Sawatsky","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6245-8813","authenticated-orcid":true,"given":"Rosal\u00eda","family":"Santos-Mandujano","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3029-9448","authenticated-orcid":true,"given":"Dominic H.","family":"Banda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Martina","family":"Oberhuber","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anika","family":"Schopf","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3298-2048","authenticated-orcid":true,"given":"Verena","family":"Pfaffinger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0027-2703","authenticated-orcid":true,"given":"Kevin","family":"Wittwer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4349-996X","authenticated-orcid":true,"given":"Christiane","family":"Riedel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3302-0780","authenticated-orcid":true,"given":"Christian K.","family":"Pfaller","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8614-3656","authenticated-orcid":true,"given":"Karl-Klaus","family":"Conzelmann","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2021,4,21]]},"reference":[{"key":"ppat.1009064.ref001","article-title":"SARS-CoV-2 mRNA vaccine design enabled by prototype pathogen preparedness","author":"KS Corbett","year":"2020","journal-title":"Nature"},{"key":"ppat.1009064.ref002","article-title":"BNT162b2 mRNA Covid-19 Vaccine in a Nationwide Mass Vaccination Setting","author":"N Dagan","year":"2021","journal-title":"The New England journal of medicine"},{"issue":"27","key":"ppat.1009064.ref003","doi-asserted-by":"crossref","first-page":"2603","DOI":"10.1056\/NEJMoa2034577","article-title":"Safety and Efficacy of the BNT162b2 mRNA Covid-19 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Cloutier","year":"2020","journal-title":"Cytokine"},{"issue":"3","key":"ppat.1009064.ref008","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1016\/j.immuni.2019.08.007","article-title":"Immunological Lessons from Respiratory Syncytial Virus Vaccine Development","volume":"51","author":"TJ Ruckwardt","year":"2019","journal-title":"Immunity"},{"issue":"6","key":"ppat.1009064.ref009","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1084\/jem.20020781","article-title":"A role for immune complexes in enhanced respiratory syncytial virus disease","volume":"196","author":"FP Polack","year":"2002","journal-title":"J Exp Med"},{"issue":"3","key":"ppat.1009064.ref010","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1128\/CVI.00609-15","article-title":"Brief History and Characterization of Enhanced Respiratory Syncytial Virus Disease","volume":"23","author":"PL Acosta","year":"2015","journal-title":"Clin Vaccine 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A"},{"key":"ppat.1009064.ref017","unstructured":"Draft landscape of COVID-19 candidate vaccines 26 March 2021 [Internet]. 2021. Available from: https:\/\/www.who.int\/publications\/m\/item\/draft-landscape-of-covid-19-candidate-vaccines."},{"key":"ppat.1009064.ref018","article-title":"Correlates of protection against SARS-CoV-2 in rhesus macaques","author":"K McMahan","year":"2020","journal-title":"Nature"},{"key":"ppat.1009064.ref019","article-title":"mRNA vaccine-elicited antibodies to SARS-CoV-2 and circulating variants","author":"Z Wang","year":"2021","journal-title":"Nature"},{"issue":"3","key":"ppat.1009064.ref020","doi-asserted-by":"crossref","first-page":"100040","DOI":"10.1016\/j.xcrm.2020.100040","article-title":"Rapid Generation of Neutralizing Antibody Responses in COVID-19 Patients","volume":"1","author":"MS Suthar","year":"2020","journal-title":"Cell reports Medicine"},{"issue":"1","key":"ppat.1009064.ref021","doi-asserted-by":"crossref","DOI":"10.1172\/jci.insight.143129","article-title":"Dissecting strategies to tune the therapeutic potential of SARS-CoV-2-specific monoclonal antibody CR3022","volume":"6","author":"C Atyeo","year":"2021","journal-title":"JCI insight."},{"key":"ppat.1009064.ref022","article-title":"Potently neutralizing and protective human antibodies against SARS-CoV-2","author":"SJ Zost","year":"2020","journal-title":"Nature"},{"issue":"9","key":"ppat.1009064.ref023","doi-asserted-by":"crossref","DOI":"10.26508\/lsa.202000786","article-title":"TMPRSS2 and furin are both essential for proteolytic activation of SARS-CoV-2 in human airway cells","volume":"3","author":"D Bestle","year":"2020","journal-title":"Life science alliance"},{"key":"ppat.1009064.ref024","article-title":"SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor","author":"M Hoffmann","year":"2020","journal-title":"Cell"},{"issue":"4","key":"ppat.1009064.ref025","doi-asserted-by":"crossref","first-page":"779","DOI":"10.1016\/j.molcel.2020.04.022","article-title":"A Multibasic Cleavage Site in the Spike Protein of SARS-CoV-2 Is Essential for Infection of Human Lung Cells","volume":"78","author":"M Hoffmann","year":"2020","journal-title":"Molecular cell"},{"issue":"7807","key":"ppat.1009064.ref026","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1038\/s41586-020-2180-5","article-title":"Structure of the SARS-CoV-2 spike receptor-binding domain bound to the ACE2 receptor","volume":"581","author":"J Lan","year":"2020","journal-title":"Nature"},{"issue":"6483","key":"ppat.1009064.ref027","doi-asserted-by":"crossref","first-page":"1260","DOI":"10.1126\/science.abb2507","article-title":"Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation","volume":"367","author":"D Wrapp","year":"2020","journal-title":"Science"},{"key":"ppat.1009064.ref028","article-title":"Characterization of the receptor-binding domain (RBD) of 2019 novel coronavirus: implication for development of RBD protein as a viral attachment inhibitor and vaccine","author":"W Tai","year":"2020","journal-title":"Cellular & molecular immunology"},{"key":"ppat.1009064.ref029","article-title":"Infection and Rapid Transmission of SARS-CoV-2 in Ferrets","author":"YI Kim","year":"2020","journal-title":"Cell host & microbe"},{"key":"ppat.1009064.ref030","article-title":"Comparative pathogenesis of COVID-19, MERS, and SARS in a nonhuman primate model","author":"B Rockx","year":"2020","journal-title":"Science"},{"key":"ppat.1009064.ref031","article-title":"Susceptibility of ferrets, cats, dogs, and other domesticated animals to SARS-coronavirus 2","author":"J Shi","year":"2020","journal-title":"Science"},{"key":"ppat.1009064.ref032","article-title":"Potent neutralizing antibodies from COVID-19 patients define multiple targets of vulnerability","author":"PJM Brouwer","year":"2020","journal-title":"Science"},{"key":"ppat.1009064.ref033","article-title":"Longitudinal Isolation of Potent Near-Germline SARS-CoV-2-Neutralizing Antibodies from COVID-19","author":"C Kreer","year":"2020","journal-title":"Patients. Cell"},{"issue":"1","key":"ppat.1009064.ref034","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/j.cell.2020.05.025","article-title":"Potent Neutralizing Antibodies against SARS-CoV-2 Identified by High-Throughput Single-Cell Sequencing of Convalescent Patients\u2019 B Cells","volume":"182","author":"Y Cao","year":"2020","journal-title":"Cell"},{"issue":"48","key":"ppat.1009064.ref035","doi-asserted-by":"crossref","DOI":"10.1126\/sciimmunol.abc8413","article-title":"The receptor binding domain of the viral spike protein is an immunodominant and highly specific target of antibodies in SARS-CoV-2 patients","volume":"5","author":"L Premkumar","year":"2020","journal-title":"Science immunology"},{"issue":"5","key":"ppat.1009064.ref036","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1021\/acsinfecdis.6b00006","article-title":"Immunodominant SARS Coronavirus Epitopes in Humans Elicited both Enhancing and Neutralizing Effects on Infection in Non-human 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biology"},{"issue":"12","key":"ppat.1009064.ref052","doi-asserted-by":"crossref","first-page":"1509","DOI":"10.1002\/jmv.20732","article-title":"Evaluation of a novel vesicular stomatitis virus pseudotype-based assay for detection of neutralizing antibody responses to SARS-CoV","volume":"78","author":"S Fukushi","year":"2006","journal-title":"Journal of medical virology"},{"key":"ppat.1009064.ref053","article-title":"A single dose of recombinant VSV-\u0394G-spike vaccine provides protection against SARS-CoV-2 challenge","author":"Y Yahalom-Ronen","year":"2020","journal-title":"bioRxiv: the preprint server for biology"},{"key":"ppat.1009064.ref054","doi-asserted-by":"crossref","DOI":"10.7554\/eLife.61312","article-title":"Escape from neutralizing antibodies by SARS-CoV-2 spike protein variants","volume":"9","author":"Y 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