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Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses.
Viruses. 2021 08 10; 13(8)V

Abstract

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.

Authors+Show Affiliations

Laboratory of Viral Zoonotics, Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UK.Laboratory of Viral Zoonotics, Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UK. DIOSynVax Ltd., Cambridge CB3 0ES, UK.Viral Pseudotype Unit, University of Kent, Chatham ME4 4TB, UK.Laboratory of Viral Zoonotics, Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UK. DIOSynVax Ltd., Cambridge CB3 0ES, UK.Laboratory of Viral Zoonotics, Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UK.Laboratory of Viral Zoonotics, Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UK.Viral Pseudotype Unit, University of Kent, Chatham ME4 4TB, UK.Viral Pseudotype Unit, University of Kent, Chatham ME4 4TB, UK.Institute for Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg, Germany.Institute for Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg, Germany.Institute for Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg, Germany. Institute for Clinical Microbiology and Hygiene, University Hospital, 93053 Regensburg, Germany.Royal Papworth Hospital NHS Foundation Trust, Cambridge CB2 0AY, UK.Viral Pseudotype Unit, University of Kent, Chatham ME4 4TB, UK.Laboratory of Viral Zoonotics, Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UK.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

34452443

Citation

Sampson, Alexander Thomas, et al. "Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses." Viruses, vol. 13, no. 8, 2021.
Sampson AT, Heeney J, Cantoni D, et al. Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses. Viruses. 2021;13(8).
Sampson, A. T., Heeney, J., Cantoni, D., Ferrari, M., Sans, M. S., George, C., Di Genova, C., Mayora Neto, M., Einhauser, S., Asbach, B., Wagner, R., Baxendale, H., Temperton, N., & Carnell, G. (2021). Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses. Viruses, 13(8). https://doi.org/10.3390/v13081579
Sampson AT, et al. Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses. Viruses. 2021 08 10;13(8) PubMed PMID: 34452443.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses. AU - Sampson,Alexander Thomas, AU - Heeney,Jonathan, AU - Cantoni,Diego, AU - Ferrari,Matteo, AU - Sans,Maria Suau, AU - George,Charlotte, AU - Di Genova,Cecilia, AU - Mayora Neto,Martin, AU - Einhauser,Sebastian, AU - Asbach,Benedikt, AU - Wagner,Ralf, AU - Baxendale,Helen, AU - Temperton,Nigel, AU - Carnell,George, Y1 - 2021/08/10/ PY - 2021/05/10/received PY - 2021/07/17/revised PY - 2021/08/01/accepted PY - 2021/8/28/entrez PY - 2021/8/29/pubmed PY - 2021/9/14/medline KW - COVID-19 KW - SARS-CoV-2 KW - coronavirus KW - neutralization KW - pseudotyped virus JF - Viruses JO - Viruses VL - 13 IS - 8 N2 - 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. SN - 1999-4915 UR - https://www.unboundmedicine.com/medline/citation/34452443/Coronavirus_Pseudotypes_for_All_Circulating_Human_Coronaviruses_for_Quantification_of_Cross_Neutralizing_Antibody_Responses_ L2 - https://www.mdpi.com/resolver?pii=v13081579 DB - PRIME DP - Unbound Medicine ER -