Therapeutic activity of an inhaled potent SARS-CoV-2 neutralizing human monoclonal antibody in hamsters

Piepenbrink, Michael S.; Park, Jun-Gyu; Oladunni, Fatai S.; Deshpande, Ashlesha; Basu, Madhubanti; Sarkar, Sanghita; Loos, Andreas; Woo, Jennifer; Lovalenti, Phillip M.; Sloan, Derek D.; Ye, Chengjin; Chiem, Kevin; Erdmann, Nathaniel; Goepfert, Paul; Truong, Vu; Walter, Mark R.; Martínez-Sobrido, Luis; Kobie, James J.

doi:10.1101/2020.10.14.339150

Cited by 21 publications

(48 citation statements)

References 40 publications

(18 reference statements)

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“…In this study, we demonstrate that reporter-expressing rSARS-CoV-2 represent an excellent option for the rapid identification and characterization of both antivirals ( Figure 4) and NAbs ( Figure 5) for the therapeutic and/or prophylactic treatment of SARS-CoV-2 infections. Importantly, EC 50 (antivirals) and NT 50 (NAbs) obtained with our reporterexpressing viruses were comparable to those obtained using rSARS-CoV-2/WT or described by others in the literature (20,23,29), demonstrating the feasibility of using our reporter-based microneutralization assays for the rapid identification of antivirals or NAbs (Figures 4 and 5, respectively).…”

Section: Discussionsupporting

confidence: 81%

“…We next evaluate the feasibility of using our reporter-expressing rSARS-CoV in a reporter-based microneutralization assay to identify NAbs against SARS-CoV-2. As proof of concept, we used a human MAb (1212C2) which we have recently described to potently bind and neutralize SARS-CoV-2 infection both in vitro and in vivo (29). The NT 50 of 1212C2 against rSARS-CoV-2-Venus ( Figures 5A and 5B, 1.94 ng), -mCherry ( Figures 5C and 5D, 5.02 ng), or Nluc (Figure 5E, 3.67 ng) were similar to those observed with rSARS-CoV-2/WT ( Figure 5F, 4.88 ng), and recently reported values (29).…”

Section: A Reporter-based Microneutralization Assay For the Identificmentioning

confidence: 99%

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Generation and Characterization of Recombinant SARS-CoV-2 Expressing Reporter Genes

Chiem

Vasquez

Park

et al. 2021

J Virol

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The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the pathogen responsible of coronavirus disease 2019 (COVID-19), has devastated public health services and economies worldwide. Despite global efforts to contain the COVID-19 pandemic, SARS-CoV-2 is now found in over 200 countries and has caused an upward death toll of over 1 million human lives as of November 2020. To date, only one Food and Drug Administration (FDA)-approved therapeutic drug (Remdesivir) and a monoclonal antibody, MAb (Bamlanivimab) are available for the treatment of SARS-CoV-2. As with other viruses, studying SARS-CoV-2 requires the use of secondary approaches to detect the presence of the virus in infected cells. To overcome this limitation, we have generated replication-competent recombinant (r)SARS-CoV-2 expressing fluorescent (Venus or mCherry) or bioluminescent (Nluc) reporter genes. Vero E6 cells infected with reporter-expressing rSARS-CoV-2 can be easily detected via fluorescence or luciferase expression and display a good correlation between reporter gene expression and viral replication. Moreover, rSARS-CoV-2 expressing reporter genes have comparable plaque sizes and growth kinetics to those of wild-type virus, rSARS-CoV-2/WT. We used these reporter-expressing rSARS-CoV-2 to demonstrate their feasibility to identify neutralizing antibodies (NAbs) or antiviral drugs. Our results demonstrate that reporter-expressing rSARS-CoV-2 represent an excellent option to identify therapeutics for the treatment of SARS-CoV-2, where reporter gene expression can be used as valid surrogates to track viral infection. Moreover, the ability to manipulate the viral genome opens the feasibility of generating viruses expressing foreign genes for their use as vaccines for the treatment of SARS-CoV-2 infection. IMPORTANCE Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the pathogen that causes coronavirus disease 2019 (COVID-19), has significantly impacted the human health and economic status worldwide. There is an urgent need to identify effective prophylactics and therapeutics for the treatment of SARS-CoV-2 infection and associated COVID-19 disease. The use of fluorescent- or luciferase-expressing reporter expressing viruses has significantly advanced viral research. Here, we generated recombinant (r)SARS-CoV-2 expressing fluorescent (Venus and mCherry) or luciferase (Nluc) reporter genes and demonstrate that they represent an excellent option to track viral infections in vitro. Importantly, reporter-expressing rSARS-CoV-2 display similar growth kinetics and plaque phenotype that their wild-type counterpart (rSARS-CoV-2/WT), demonstrating their feasibility to identify drugs and/or neutralizing antibodies (NAbs) for the therapeutic treatment of SARS-CoV-2. Henceforth, these reporter-expressing rSARS-CoV-2 can be used to interrogate large libraries of compounds and/or monoclonal antibodies (MAb), in high-throughput screening settings, to identify those with therapeutic potential against SARS-CoV-2.

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Section: Discussionsupporting

confidence: 81%

Section: A Reporter-based Microneutralization Assay For the Identificmentioning

confidence: 99%

Generation and Characterization of Recombinant SARS-CoV-2 Expressing Reporter Genes

Chiem

Vasquez

Park

et al. 2021

J Virol

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“…To test the neutralizing activity of 1212C2, a human MAb recently described to neutralize SARS-CoV-2 (27), confluent monolayers of Vero E6 cells (96-plate format, 4 × 10 4 cells/well, quadruplicates) were infected with ~100-200 PFU of rSARS-CoV-2/WT or rSARS-CoV-2 expressing Venus, mCherry, or Nluc for 1 h at 37°C. After viral adsorption, cells were washed and incubated with 100 μL of infection media (DMEM 2% FBS) containing 3-fold serial dilutions (starting concentration of 500 ng) of 1212C2 or PBS, and 1% avicel (Sigma-Aldrich).…”

Section: Methodsmentioning

confidence: 99%

Generation and Characterization of recombinant SARS-CoV-2 expressing reporter genes

Chiem

Vasquez

Park

et al. 2020

Preprint

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The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the pathogen responsible of coronavirus disease 2019 (COVID-19), has devastated public health services and economies worldwide. Despite global efforts to contain the COVID-19 pandemic, SARS-CoV-2 is now found in over 200 countries and has caused an upward death toll of over 1 million human lives as of November 2020. To date, only one Food and Drug Administration (FDA)-approved therapeutic drug (Remdesivir) and a monoclonal antibody, MAb (Bamlanivimab), but no vaccines, are available for the treatment of SARS-CoV-2. As with other viruses, studying SARS-CoV-2 requires the use of secondary approaches to detect the presence of the virus in infected cells. To overcome this limitation, we have generated replication-competent recombinant (r)SARS-CoV-2 expressing fluorescent (Venus or mCherry) or bioluminescent (Nluc) reporter genes. Vero E6 cells infected with reporter-expressing rSARS-CoV-2 can be easily detected via fluorescence or luciferase expression and display a good correlation between reporter gene expression and viral replication. Moreover, rSARS-CoV-2 expressing reporter genes have comparable plaque sizes and growth kinetics to those of wild-type virus, rSARS-CoV-2/WT. We used these reporter-expressing rSARS-CoV-2 to demonstrate their feasibility to identify neutralizing antibodies (NAbs) or antiviral drugs. Our results demonstrate that reporter-expressing rSARS-CoV-2 represent an excellent option to identify therapeutics for the treatment of SARS-CoV-2, where reporter gene expression can be used as valid surrogates to track viral infection. Moreover, the ability to manipulate the viral genome opens the feasibility of generating viruses expressing foreign genes for their use as vaccines for the treatment of SARS-CoV-2 infection.ImportanceSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the pathogen that causes coronavirus disease 2019 (COVID-19), has significantly impacted the human health and economic status worldwide. There is an urgent need to identify effective prophylactics and therapeutics for the treatment of SARS-CoV-2 infection and associated COVID-19 disease. The use of fluorescent- or luciferase-expressing reporter expressing viruses has significantly advanced viral research. Here, we generated recombinant (r)SARS-CoV-2 expressing fluorescent (Venus and mCherry) or luciferase (Nluc) reporter genes and demonstrate that they represent an excellent option to track viral infections in vitro. Importantly, reporter-expressing rSARS-CoV-2 display similar growth kinetics and plaque phenotype that their wild-type counterpart (rSARS-CoV-2/WT), demonstrating their feasibility to identify drugs and/or neutralizing antibodies (NAbs) for the therapeutic treatment of SARS-CoV-2. Henceforth, these reporter-expressing rSARS-CoV-2 can be used to interrogate large libraries of compounds and/or monoclonal antibodies (MAb), in high-throughput screening settings, to identify those with therapeutic potential against SARS-CoV-2.

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“…These neutralizing antibodies are targeting the spike protein, but they bind to different epitopes where some are engineered regarding effector functions and FcRn binding. In addition, delivery of neutralizing IgG antibodies to the lungs via inhalation may also be an attractive approach as to improve bioavailability and dosing efficiency 70 …”

Section: Main Textmentioning

confidence: 99%

“…In addition, delivery of neutralizing IgG antibodies to the lungs via inhalation may also be an attractive approach as to improve bioavailability and dosing efficiency. 70…”

Section: The Protective Role Of Fcrn Upon Viral and Bacterial Infecmentioning

confidence: 99%

The neonatal Fc receptor in mucosal immune regulation

et al. 2021

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The neonatal Fc receptor (FcRn) was first recognized for its role in transfer of maternal IgG to the foetus or newborn, providing passive immunity early in life. However, it has become clear that the receptor is versatile, widely expressed and plays an indispensable role in both immunological and non-immunological processes throughout life. The receptor rescues immunoglobulin G (IgG) and albumin from intracellular degradation and shuttles the ligands across polarized cell barriers, in all cases via a pH-dependent binding-and-release mechanism. These processes secure distribution and high levels of both IgG and albumin throughout the body. At mucosal sites, FcRn transports IgG across polarized epithelial cells where it retrieves IgG in complex with luminal antigens that is delivered to tissue-localized immune cells. In dendritic cells (DCs), FcRn orchestrates processing of IgG-opsonized immune complexes (ICs) in concert with classical Fcγ receptors, which results in antigen presentation and cross-presentation of antigenic peptides on MHC class II and I to CD4+ and CD8+ T cells, respectively. Hence, FcRn regulates transport of the ligands within and across different types of cells, but also processing of IgG-ICs by immune cells. As such, the receptor is involved in immune surveillance and protection against infections. In this brief review, we highlight how FcRn expressed by hematopoietic and non-hematopoietic cells contributes to immune regulation at mucosal barriersbiology that can be utilized in development of biologics and subunit vaccines for non-invasive delivery.

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Therapeutic activity of an inhaled potent SARS-CoV-2 neutralizing human monoclonal antibody in hamsters

Cited by 21 publications

References 40 publications

Generation and Characterization of Recombinant SARS-CoV-2 Expressing Reporter Genes

Generation and Characterization of Recombinant SARS-CoV-2 Expressing Reporter Genes

Generation and Characterization of recombinant SARS-CoV-2 expressing reporter genes

The neonatal Fc receptor in mucosal immune regulation

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