Abstract
BackgroundThe spread of SARS-CoV-2, the virus that causes Coronavirus Disease 2019 (COVID-19), has been characterized as a worldwide pandemic. Currently, there are few preclinical animal models that suitably represent infection, as the main point of entry to human cells is via human angiotensin-converting enzyme 2 (ACE2) which is not present in typical preclinical mouse strains. Additionally, SARS-CoV-2 is highly virulent and unsafe for use in many research facilities. Here we describe the development of a preclinical animal model using intranasal administration of ACE2 followed by non-infectious SARS-CoV-2 pseudovirus (PsV) challenge.MethodsTo specifically generate our SARS-CoV-2 PsV, we used a lentivirus system. Following co-transfection with a packaging plasmid containing HIV Gag and Pol, luciferase-expressing lentiviruses, and a plasmid carrying the SARS-CoV-2 spike protein, SARS-CoV-2 PsVs can be isolated and purified. To better understand and maximize the infectivity of SARS-CoV-2 PsV, we generated PsV carrying spike protein variants known to have varying human ACE2 binding properties, including 19 deletion (19del) and 19del + D614G.ResultsOur system demonstrated the ability of PsVs to infect the respiratory passage of mice following intranasal hACE2 transduction. Additionally, we demonstrate in vitro and in vivo manipulability of our system using recombinant receptor-binding domain protein to prevent PsV infection.ConclusionsOur PsV system is able to model SARS-CoV-2 infections in a preclinical mouse model and can be used to test interventions or preventative treatments. We believe that this method can be extended to work in various mouse strains or to model infection with different coronaviruses. A simple in vivo system such as our model is crucial for rapidly and effectively responding to the current COVID-19 pandemic in addition to preparing for future potential coronavirus outbreaks.
Highlights
Since December 2019, when the World Health Organization first cited the disease, SARS-CoV-2 has spread to219 countries
Mice infected intranasally with SARS‐CoV‐2 pseudovirus post Adenovirus vector (AdV)‐Human ACE2 (hACE2) demonstrate signs of infection We first sought to assess the ability of our SARS-CoV-2 PsVs to infect cells expressing hACE2. 293TT cells expressing hACE2 were infected with WT, 19del, or 19del + D614G PsVs. 72 h later, cells were lysed, and luminescence activity was measured using a luminometer
We observed that WT SARS-CoV-2 PsVs exhibit a ~ 30-fold increase in relative luminescence units (RLU) (Fig. 1a), indicating that substantial infection occurred
Summary
Since December 2019, when the World Health Organization first cited the disease, SARS-CoV-2 has spread to219 countries. Since December 2019, when the World Health Organization first cited the disease, SARS-CoV-2 has spread to. Coronavirus Disease 2019 (COVID-19) has been characterized as a pandemic. The severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) is the causative agent of COVID-19. SARS-CoV-2, as well as other related coronavirus family members, utilize host angiotensin-converting enzyme 2 (ACE2) as the binding and entry receptor [3]. The receptorbinding domain (RBD) of SARS-CoV-2, as with SARSCoV, permits binding to ACE2, and is located in S1 [5]. The spread of SARS-CoV-2, the virus that causes Coronavirus Disease 2019 (COVID-19), has been characterized as a worldwide pandemic. We describe the development of a preclinical animal model using intranasal administration of ACE2 followed by non-infectious SARS-CoV-2 pseudovirus (PsV) challenge
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