Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses
The 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 t...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-08-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/13/8/1579 |
| _version_ | 1797521828925669376 |
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| author | Alexander Thomas Sampson Jonathan Heeney Diego Cantoni Matteo Ferrari Maria Suau Sans Charlotte George Cecilia Di Genova Martin Mayora Neto Sebastian Einhauser Benedikt Asbach Ralf Wagner Helen Baxendale Nigel Temperton George Carnell |
| author_facet | Alexander Thomas Sampson Jonathan Heeney Diego Cantoni Matteo Ferrari Maria Suau Sans Charlotte George Cecilia Di Genova Martin Mayora Neto Sebastian Einhauser Benedikt Asbach Ralf Wagner Helen Baxendale Nigel Temperton George Carnell |
| author_sort | Alexander Thomas Sampson |
| collection | DOAJ |
| description | The 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. |
| first_indexed | 2024-03-10T08:19:04Z |
| format | Article |
| id | doaj.art-f0dbf2fae91a450ab1b663d93e579583 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T08:19:04Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-f0dbf2fae91a450ab1b663d93e5795832023-11-22T10:11:35ZengMDPI AGViruses1999-49152021-08-01138157910.3390/v13081579Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody ResponsesAlexander Thomas Sampson0Jonathan Heeney1Diego Cantoni2Matteo Ferrari3Maria Suau Sans4Charlotte George5Cecilia Di Genova6Martin Mayora Neto7Sebastian Einhauser8Benedikt Asbach9Ralf Wagner10Helen Baxendale11Nigel Temperton12George Carnell13Laboratory of Viral Zoonotics, Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UKLaboratory of Viral Zoonotics, Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UKViral Pseudotype Unit, University of Kent, Chatham ME4 4TB, UKLaboratory of Viral Zoonotics, Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UKLaboratory of Viral Zoonotics, Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UKLaboratory of Viral Zoonotics, Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UKViral Pseudotype Unit, University of Kent, Chatham ME4 4TB, UKViral Pseudotype Unit, University of Kent, Chatham ME4 4TB, UKInstitute for Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg, GermanyInstitute for Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg, GermanyInstitute for Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg, GermanyRoyal Papworth Hospital NHS Foundation Trust, Cambridge CB2 0AY, UKViral Pseudotype Unit, University of Kent, Chatham ME4 4TB, UKLaboratory of Viral Zoonotics, Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UKThe 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.https://www.mdpi.com/1999-4915/13/8/1579SARS-CoV-2COVID-19coronaviruspseudotyped virusneutralization |
| spellingShingle | Alexander Thomas Sampson Jonathan Heeney Diego Cantoni Matteo Ferrari Maria Suau Sans Charlotte George Cecilia Di Genova Martin Mayora Neto Sebastian Einhauser Benedikt Asbach Ralf Wagner Helen Baxendale Nigel Temperton George Carnell Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses Viruses SARS-CoV-2 COVID-19 coronavirus pseudotyped virus neutralization |
| title | Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses |
| title_full | Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses |
| title_fullStr | Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses |
| title_full_unstemmed | Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses |
| title_short | Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses |
| title_sort | coronavirus pseudotypes for all circulating human coronaviruses for quantification of cross neutralizing antibody responses |
| topic | SARS-CoV-2 COVID-19 coronavirus pseudotyped virus neutralization |
| url | https://www.mdpi.com/1999-4915/13/8/1579 |
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