Ssu-Hsueh Tseng1, Brandon Lam1,2, Yu Jui Kung1, John Lin1, Li Liu1, Ya Chea Tsai1, Louise Ferrall1, Richard B S Roden1,3,4, T C Wu1,3,4,5, Chien-Fu Hung6,7,8. 1. Department of Pathology, Johns Hopkins School of Medicine, Baltimore, MD, USA. 2. Graduate Program in Immunology, Johns Hopkins School of Medicine, Baltimore, MD, USA. 3. Department of Oncology, Johns Hopkins School of Medicine, Baltimore, MD, USA. 4. Department of Obstetrics and Gynecology, Johns Hopkins School of Medicine, Baltimore, MD, USA. 5. Molecular Microbiology and Immunology, Johns Hopkins School of Medicine, Baltimore, MD, USA. 6. Department of Pathology, Johns Hopkins School of Medicine, Baltimore, MD, USA. chung2@jhmi.edu. 7. Department of Oncology, Johns Hopkins School of Medicine, Baltimore, MD, USA. chung2@jhmi.edu. 8. Johns Hopkins University School of Medicine, 1550 Orleans Street, CRBII 307, Baltimore, MD, 21287, USA. chung2@jhmi.edu.
Abstract
BACKGROUND: The 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. METHODS: To 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. RESULTS: Our 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. CONCLUSIONS: Our 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.
BACKGROUND: The 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 humanangiotensin-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. METHODS: To 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-2spike 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 humanACE2 binding properties, including 19 deletion (19del) and 19del + D614G. RESULTS: Our 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. CONCLUSIONS: Our 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.
Authors: Yaozong Chen; Lulu Sun; Irfan Ullah; Guillaume Beaudoin-Bussières; Sai Priya Anand; Andrew P Hederman; William D Tolbert; Rebekah Sherburn; Dung N Nguyen; Lorie Marchitto; Shilei Ding; Di Wu; Yuhong Luo; Suneetha Gottumukkala; Sean Moran; Priti Kumar; Grzegorz Piszczek; Walther Mothes; Margaret E Ackerman; Andrés Finzi; Pradeep D Uchil; Frank J Gonzalez; Marzena Pazgier Journal: Sci Adv Date: 2022-07-13 Impact factor: 14.957
Authors: Yaozong Chen; Lulu Sun; Irfan Ullah; Guillaume Beaudoin-Bussières; Sai Priya Anand; Andrew P Hederman; William D Tolbert; Rebekah Sherburn; Dung N Nguyen; Lorie Marchitto; Shilei Ding; Di Wu; Yuhong Luo; Suneetha Gottumukkala; Sean Moran; Priti Kumar; Grzegorz Piszczek; Walther Mothes; Margaret E Ackerman; Andrés Finzi; Pradeep D Uchil; Frank J Gonzalez; Marzena Pazgier Journal: bioRxiv Date: 2021-11-24