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dc.contributor.authorSampson, Alexander Thomas
dc.contributor.authorHeeney, Jonathan
dc.contributor.authorCantoni, Diego
dc.contributor.authorFerrari, Matteo
dc.contributor.authorSans, Maria Suau
dc.contributor.authorGeorge, Charlotte
dc.contributor.authorDi Genova, Cecilia
dc.contributor.authorMayora Neto, Martin
dc.contributor.authorEinhauser, Sebastian
dc.contributor.authorAsbach, Benedikt
dc.contributor.authorWagner, Ralf
dc.contributor.authorBaxendale, Helen
dc.contributor.authorTemperton, Nigel
dc.contributor.authorCarnell, George
dc.date.accessioned2021-10-25T23:30:34Z
dc.date.available2021-10-25T23:30:34Z
dc.date.issued2021-08-10
dc.identifier.issn1999-4915
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/329866
dc.description.abstractThe novel coronavirus SARS-CoV-2 is the seventh identified human coronavirus. Understanding the extent of pre-existing immunity induced by seropositivity to endemic seasonal coronaviruses and the impact of cross-reactivity on COVID-19 disease progression remains a key research question in immunity to SARS-CoV-2 and the immunopathology of COVID-2019 disease. This paper describes a panel of lentiviral pseudotypes bearing the spike (S) proteins for each of the seven human coronaviruses (HCoVs), generated under similar conditions optimized for high titre production allowing a high-throughput investigation of antibody neutralization breadth. Optimal production conditions and most readily available permissive target cell lines were determined for spike-mediated entry by each HCoV pseudotype: SARS-CoV-1, SARS-CoV-2 and HCoV-NL63 best transduced HEK293T/17 cells transfected with ACE2 and TMPRSS2, HCoV-229E and MERS-CoV preferentially entered HUH7 cells, and CHO cells were most permissive for the seasonal betacoronavirus HCoV-HKU1. Entry of ACE2 using pseudotypes was enhanced by ACE2 and TMPRSS2 expression in target cells, whilst TMPRSS2 transfection rendered HEK293T/17 cells permissive for HCoV-HKU1 and HCoV-OC43 entry. Additionally, pseudotype viruses were produced bearing additional coronavirus surface proteins, including the SARS-CoV-2 Envelope (E) and Membrane (M) proteins and HCoV-OC43/HCoV-HKU1 Haemagglutinin-Esterase (HE) proteins. This panel of lentiviral pseudotypes provides a safe, rapidly quantifiable and high-throughput tool for serological comparison of pan-coronavirus neutralizing responses; this can be used to elucidate antibody dynamics against individual coronaviruses and the effects of antibody cross-reactivity on clinical outcome following natural infection or vaccination.
dc.format.mediumElectronic
dc.languageeng
dc.publisherMDPI AG
dc.rightsAttribution 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectCell Line
dc.subjectAnimals
dc.subjectHumans
dc.subjectCoronavirus
dc.subjectCoronavirus 229E, Human
dc.subjectCoronavirus OC43, Human
dc.subjectLentivirus
dc.subjectAntibodies, Viral
dc.subjectNeutralization Tests
dc.subjectTransfection
dc.subjectCross Reactions
dc.subjectPlasmids
dc.subjectVirus Internalization
dc.subjectCoronavirus NL63, Human
dc.subjectSpike Glycoprotein, Coronavirus
dc.subjectMiddle East Respiratory Syndrome Coronavirus
dc.subjectBroadly Neutralizing Antibodies
dc.subjectCOVID-19
dc.subjectSARS-CoV-2
dc.titleCoronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses.
dc.typeArticle
prism.issueIdentifier8
prism.publicationDate2021
prism.publicationNameViruses
prism.volume13
dc.identifier.doi10.17863/CAM.77311
dcterms.dateAccepted2021-08-01
rioxxterms.versionofrecord10.3390/v13081579
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2021-08-10
dc.contributor.orcidHeeney, Jonathan [0000-0003-2702-1621]
dc.contributor.orcidDi Genova, Cecilia [0000-0002-2121-0715]
dc.contributor.orcidEinhauser, Sebastian [0000-0003-4177-4451]
dc.contributor.orcidAsbach, Benedikt [0000-0003-1056-8591]
dc.contributor.orcidTemperton, Nigel [0000-0002-7978-3815]
dc.contributor.orcidCarnell, George [0000-0001-8875-0989]
dc.identifier.eissn1999-4915
rioxxterms.typeJournal Article/Review
pubs.funder-project-idInnovate UK (971616)
cam.issuedOnline2021-08-10


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International