K18-hACE2 Mice for Studies of COVID-19 Treatments and Pathogenesis Including Anosmia
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Abstract
The ongoing COVID-19 pandemic is associated with substantial morbidity and mortality. While much has been learned in the first months of the pandemic, many features of COVID-19 pathogenesis remain to be determined. For example, anosmia is a common presentation and many patients with this finding show no or only minor respiratory signs. Studies in animals experimentally infected with SARS-CoV-2, the cause of COVID-19, provide opportunities to study aspects of the disease not easily investigated in human patients. COVID-19 severity ranges from asymptomatic to lethal. Most experimental infections provide insights into mild disease. Here, using K18-hACE2 mice that we originally developed for SARS studies, we show that infection with SARS-CoV-2 causes severe disease in the lung, and in some mice, the brain. Evidence of thrombosis and vasculitis was detected in mice with severe pneumonia. Further, we show that infusion of convalescent plasma (CP) from a recovered COVID-19 patient provided protection against lethal disease. Mice developed anosmia at early times after infection. Notably, while treatment with CP prevented significant clinical disease, it did not prevent anosmia. Thus K18-hACE2 mice provide a useful model for studying the pathological underpinnings of both mild and lethal COVID-19 and for assessing therapeutic interventions.
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SciScore for 10.1101/2020.08.07.242073: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: Human subjects approval: Written informed consent was obtained from subjects to obtain plasma for participation in this study.
IRB: The study was approved by the Institutional Review Board of the University of Iowa (IRB (#202003554 and #201402735)
IACUC: All protocols were approved by the Institutional Animal Care and Use Committees of the University of Iowa.Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable The convalescent plasma donor was a 58 year old female who had molecularly confirmed COVID-19 more than 4 weeks prior to their donation. Cell Line Authentication not detected. Table 2: Resources
Antibodies SciScore for 10.1101/2020.08.07.242073: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: Human subjects approval: Written informed consent was obtained from subjects to obtain plasma for participation in this study.
IRB: The study was approved by the Institutional Review Board of the University of Iowa (IRB (#202003554 and #201402735)
IACUC: All protocols were approved by the Institutional Animal Care and Use Committees of the University of Iowa.Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable The convalescent plasma donor was a 58 year old female who had molecularly confirmed COVID-19 more than 4 weeks prior to their donation. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Flow Cytometry: The following monoclonal antibodies were used: anti-mouse CD3e-BV421 (clone 145-2C11, Cat. No.: 562600) anti-mouse CD3e-BV421suggested: None, anti-mouse IFN-γ-APC (clone XMG1.2, Cat. No.: 25-7311-82), eBioscience, San Diego, CA; and mouse anti-C9 antibody (clone Rho 1D4, Cat. No.: MAB5356 anti-mouse IFN-γ-APCsuggested: Noneanti-C9suggested: NoneCells were then labeled for cell-surface markers, fixed/permeabilized with Cytofix/Cytoperm Solution (BD Biosciences), and labeled with anti-IFN-γ and anti-TNF antibody. anti-IFN-γsuggested: Noneanti-TNFsuggested: NoneFor SARS-CoV-2 antigen detection, slides were incubated with blocking reagent (10% normal goat serum x 30 minutes) followed by rabbit monoclonal antibody against SARS-CoV2 N protein (1:20,000 dilution x 60 minutes, #40143-R019, Sino Biological US Inc., Wayne, PA, USA), then incubated with Rabbit Envision (Dako) and diaminobenzidine (Dako) as chromogen. SARS-CoV2 N proteinsuggested: NonePseudovirus neutralizing antibody assay: To determine neutralization activity of patient and mouse plasma, we used a luciferase reporter-based pseudovirus neutralization assay, which has a nonreplicative vesicular stomatitis virus backbone coated with the SARS-CoV-2 spike protein. SARS-CoV-2 spike protein.suggested: NoneExperimental Models: Cell Lines Sentences Resources The 2019n-CoV/USA-WA1/2019 strain of SARS-CoV-2 (Accession number: MT985325.1) used in these studies was passaged on Calu-3 2B4 cells. Calu-3 2B4suggested: RRID:CVCL_YZ47)Samples were placed onto Vero E6 cells and incubated at 37°C for 1 h to allow virus binding. Vero E6suggested: NoneExperimental Models: Organisms/Strains Sentences Resources Non-transgenic C57BL/6 mice were used as controls in some experiments. C57BL/6suggested: NoneSoftware and Algorithms Sentences Resources All flow cytometry data were acquired using a BD FACSVerse and analyzed with FlowJo software. FlowJosuggested: (FlowJo, RRID:SCR_008520)Behavioral studies: Statistical analysis: ANOVA and Student’s t tests were used to analyze differences in mean values between groups using GraphPad Prism 7. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
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SciScore for 10.1101/2020.08.07.242073: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
No key resources detected.
Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.
Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different …
SciScore for 10.1101/2020.08.07.242073: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
No key resources detected.
Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.
Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
Results from JetFighter: We did not find any issues relating to colormaps.
About SciScore
SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.
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