A single intranasal dose of chimpanzee adenovirus-vectored vaccine confers sterilizing immunity against SARS-CoV-2 infection

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   ScreenIT

   Jul 17, 2020

   SciScore for 10.1101/2020.07.16.205088: (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.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Cells were washed then sequentially incubated with anti-SARS-CoV-2 CR3022 antibody (Yuan et al., 2020) (1 μg/mL) and a HRP-conjugated goat anti-human IgG (Sigma) in PBS supplemented with 0.1% (w/v) saponin (Sigma) and 0.1% BSA.	anti-SARS-CoV-2 CR3022 suggested: (Abcam Cat# ab273074, RRID:AB_2847846) anti-human IgG suggested: None
   Subsequently, plates were washed with PBS and incubated overnight at 4°C with biotinylated anti-HuAdV5-hexon antibody (2 µg/mL; …

   SciScore for 10.1101/2020.07.16.205088: (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.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Cells were washed then sequentially incubated with anti-SARS-CoV-2 CR3022 antibody (Yuan et al., 2020) (1 μg/mL) and a HRP-conjugated goat anti-human IgG (Sigma) in PBS supplemented with 0.1% (w/v) saponin (Sigma) and 0.1% BSA.	anti-SARS-CoV-2 CR3022 suggested: (Abcam Cat# ab273074, RRID:AB_2847846) anti-human IgG suggested: None
   Subsequently, plates were washed with PBS and incubated overnight at 4°C with biotinylated anti-HuAdV5-hexon antibody (2 µg/mL; Novus Biologicals NB600413) diluted in permeabilization buffer (PBS supplemented with 0.1% (w/v) saponin and 0.1% BSA).	anti-HuAdV5-hexon suggested: None
   Following blocking with FcγR antibody (BioLegend, clone 93), cells were stained on ice with CD45 BUV395 (BD BioSciences clone 30-F11); CD44 PE-Cy7, CD4 PE-Cy5, CD8b PreCP-Cy5.5, and CD19 APC-Cy7 (BioLegend clones, IM7, GK1.5, YTS156.7.7, and 6D5, respectively), and Fixable Aqua Dead Cell Stain (Invitrogen, L34966)	CD45 suggested: None CD44 suggested: None CD19 suggested: None APC-Cy7 suggested: None L34966 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   The rescued replication-incompetent ChAd-SARS-CoV-2-S and ChAd-Control vectors were scaled up in 293 cells and purified by CsCl density-gradient ultracentrifugation.	293 suggested: NCI-DTP Cat# NCI-293TT, RRID:CVCL_1D85)
   The pAd-hACE2 was linearized with PacI restriction enzyme before transfection into T-Rex 293 HEK cells (Invitrogen) to generate	HEK suggested: CLS Cat# 300192/p777_HEK293, RRID:CVCL_0045)
   The viral titer was determined by plaque assay in 293-HEK cells.	293-HEK suggested: None
   The virus-serum mixtures were added to Vero cell monolayers in 96-well plates and incubated for 1 h at 37°C.	Vero suggested: None
   Vero E6 cells plated in clear-bottom black-walled 96-well plates (Corning) were inoculated with serum-virus mixtures, and cells were cultured at 37°C for 48 h.	Vero E6 suggested: None
   The virus-serum mixtures were added to HEK293 cell monolayers in 96-well plates and incubated for 1 h at 37°C.	HEK293 suggested: None
   Flow cytometry-based antigen characterization: HEK-293T cells were seeded at 106 cells/well in 6-well plates 24 h prior to transduction with ChAd-SARS-CoV-2-S (MOI of 5).	HEK-293T suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Protein expression and purification: Purified RNA from the 2019-nCoV/USA-WA1/2020 SARS-CoV-2 strain was reverse transcribed into cDNA and used as the template for recombinant gene cloning.	SARS-CoV-2 suggested: None
   Software and Algorithms
   Sentences	Resources
   Following a 1 h incubation at room temperature, plates were washed thrice with PBST and 50 µL of 1-Step Ultra TMB-ELISA was added (ThermoFisher Cat. #34028).	ThermoFisher Cat. suggested: None
   Analysis was performed on a BD LSRFortessa X-20 cytometer, using FlowJo X 10.0 software.	FlowJo suggested: (FlowJo, RRID:SCR_008520)

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   Results from OddPub: Thank you for sharing your data.

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   Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:

   One limitation of these challenge models is that rhesus macaques develop mild interstitial pneumonia after SARS-CoV-2 infection compared to some humans and other non-human primate species (Chandrashekar et al., 2020; Munster et al., 2020). Our study in hACE2-expressing mice show that a single intramuscular or intranasal dose of ChAd-SARS-CoV-2-S vaccine confers substantial and possibly complete protection against viral replication, inflammation, and lung disease. Our approach supports the use of non-human Ad-vectored vaccines against emerging RNA viruses including SARS-CoV-2. Previously, we showed the efficacy of single-dose or two-dose regimens of a gorilla Ad encoding the prM-E genes of Zika virus (ZIKV) in several mouse challenge models including in the context of pregnancy (Hassan et al., 2019). Others have evaluated ChAd or rhesus macaque Ad vaccine candidates against ZIKV and shown efficacy in mice and non-human primates (Abbink et al., 2016; López-Camacho et al., 2018). A different ChAd encoding the wild-type SARS-CoV-2 S protein (ChAdOx1) is currently in clinical trials in humans (NCT04324606). Although data from the human trials has not yet been reported, studies in rhesus macaques suggest that a single intramuscular dose protects against infection in the lung but not in the upper respiratory tract (https://www.biorxiv.org/content/10.1101/2020.05.13.093195v1). None of the vaccines evaluated against SARS-CoV-2 has shown evidence of immune enhancement in any pre-clinic...

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   Results from TrialIdentifier: We found the following clinical trial numbers in your paper:
   

   Identifier	Status	Title
   NCT04324606	Active, not recruiting	A Study of a Candidate COVID-19 Vaccine (COV001)

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   Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

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   Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on pages 40, 43 and 37. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

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   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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   Version published to 10.1101/2020.07.16.205088 on bioRxiv

   Jul 17, 2020