TY - JOUR
AU - Sampson, Alexander Thomas
AU - Heeney, Jonathan L.
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 W.
PY - 2021
DA - August
TI - Coronavirus pseudotypes for all circulating human coronaviruses for quantification of cross-neutralizing antibody responses
JO - Viruses
VL - 13
IS - 8
DO - https://doi.org/10.3390/v13081579
AB - 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.
PB - MDPI
UR - https://eprints.gla.ac.uk/308365/
KW - Coronavirus (COVID-19)
ER