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.
Highlights
Since its emergence in 2019 [1], the novel coronavirus SARS-CoV-2 (Severe AcuteRespiratory Syndrome Coronavirus 2) has been responsible for over 190 million cases of Coronavirus Disease 2019 (COVID-19) worldwide
This paper presents a panel of lentiviral PVs pseudotyped with S proteins from each of the seven human coronaviruses
We demonstrated that our panel can be used to investigate the neutralization of different HCoVs by human serum samples—. This could be used to characterise an individual’s ability to neutralize multiple human coronaviruses (Figure 9a), to compare the neutralization ability between sub-groups in a cohort of serum samples (Figure 9b) or to study changes in the neutralization ability over time (Figure 10). This allows a further investigation of responses to individual HCoVs, including the assessment of antibody dynamics and the longevity of protection, and can be used to investigate cross-neutralizing responses across the human coronaviruses
Summary
Since its emergence in 2019 [1], the novel coronavirus SARS-CoV-2 (Severe AcuteRespiratory Syndrome Coronavirus 2) has been responsible for over 190 million cases of Coronavirus Disease 2019 (COVID-19) worldwide. The CCCoVs are endemic, frequently first encountered in the first decade of life. They are isolated seasonally from a substantial proportion of respiratory diseases in adults, suggesting a highly prevalent re-exposure and short-lived immune protection despite a widespread seropositivity [5,6,7,8]. Throughout the SARS-CoV-2 pandemic, there have been questions regarding the extent of pre-existing immunity to SARS-CoV-2 induced by prior exposure to seasonal coronaviruses and the impact of cross-reactivity on disease progression [9]. Understanding the dynamics of antibody responses observed after the SARS-CoV-1 pandemic, MERS-CoV outbreaks or a following seasonal coronavirus infection may help define the durability of protection induced by vaccination or natural infection with SARS-CoV-2 [8]
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