Chimeric spike mRNA vaccines protect against Sarbecoviru s challenge in mice

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   ScreenIT

   Mar 15, 2021

   SciScore for 10.1101/2021.03.11.434872: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: The study was carried out in accordance with the recommendations for care and use of animals by the Office of Laboratory Animal Welfare (OLAW), National Institutes of Health and the Institutional Animal Care and Use Committee (IACUC) of University of North Carolina (UNC permit no. A-3410-01).
   Randomization	In a blinded manner, three random fields of lung tissue were chosen and scored for the following: (A) neutrophils in the alveolar space (none = 0, 1–5 cells = 1, > 5 cells = 2), (B) neutrophils in the interstitial septa (none = 0, 1–5 cells = 1, > 5 cells = 2), (C) hyaline membranes (none = 0, one membrane = 1, > 1 membrane = 2), (D) Proteinaceous debris …

   SciScore for 10.1101/2021.03.11.434872: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: The study was carried out in accordance with the recommendations for care and use of animals by the Office of Laboratory Animal Welfare (OLAW), National Institutes of Health and the Institutional Animal Care and Use Committee (IACUC) of University of North Carolina (UNC permit no. A-3410-01).
   Randomization	In a blinded manner, three random fields of lung tissue were chosen and scored for the following: (A) neutrophils in the alveolar space (none = 0, 1–5 cells = 1, > 5 cells = 2), (B) neutrophils in the interstitial septa (none = 0, 1–5 cells = 1, > 5 cells = 2), (C) hyaline membranes (none = 0, one membrane = 1, > 1 membrane = 2), (D) Proteinaceous debris in air spaces (none = 0, one instance = 1, > 1 instance = 2), (E) alveolar septal thickening (< 2x mock thickness = 0, 2–4x mock thickness = 1, > 4x mock thickness = 2).
   Blinding	Analyses and scoring were performed by a board vertified veterinary pathologist who was blinded to the treatment groups as described previously (36).
   Power Analysis	not detected.
   Sex as a biological variable	Animals, immunizations, and challenge viruses: Eleven-month-old female BALB/c mice were purchased from Envigo (#047) and were used for all experiments.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   HRP conjugated goat anti-mouse IgG secondary antibody (SouthernBiotech 1030-05) was diluted to 1:10,000 in assay diluent without azide, incubated at for 1 hour at room temperature, washed and detected with 20μl SureBlue Reserve (KPL 53-00-03) for 15 minutes.	anti-mouse IgG suggested: (SouthernBiotech Cat# 1030-05, RRID:AB_2619742)
   Antibody-virus and virus only mixtures were then incubated at 37°C with 5% CO2 for one hour.	Antibody-virus suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Virus titers were measured in Vero E6 USAMRIID cells, as defined by plaque forming units (PFU) per ml, in a 6-well plate format in quadruplicate biological replicates for accuracy.	Vero E6 suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Animals, immunizations, and challenge viruses: Eleven-month-old female BALB/c mice were purchased from Envigo (#047) and were used for all experiments.	BALB/c suggested: None
   Software and Algorithms
   Sentences	Resources
   Statistics: All statistical analyses were performed using GraphPad Prism 9.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)

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   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).

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

   A caveat of including multiple chimeric spikes in a single shot is the potential formation of heterotrimers not present in the intended vaccine formulation. While it remains unknown if our chimeric mRNA-LNP vaccines generate heterotrimers in vivo, the robustness of the cross-neutralizing titers against sarbecoviruses and protection against SARS-CoV and SARS-CoV-2 in groups 1 and 2 in aged mice lends support to this strategy as a way to elicit broadly cross-reactive neutralizing antibodies against Group 2B coronaviruses. In agreement with this notion, chimera 4, which contains the RsSHC014 RBD and SARS-CoV-2 NTD and S2, elicited binding and neutralizing antibodies and also fully protected mice from BtCoV RsSHC014 and SARS-CoV-2 challenge, suggesting that CoV spikes vaccines can be designed to maximize their display of neutralizing and protective epitopes that can cover more than one pandemic/epidemic/pre-emergent CoV that are at high risk for emergence into naïve human populations. While other strategies also exist, such as multiplexing mosaic sarbecovirus RBDs (19), S1 or spike glycoproteins and RBDs on nanoparticles (45), chimeric spike mRNA-LNP vaccination can clearly achieve broad protection, using existing manufacturing technologies, and are highly portable to other high-risk emerging coronaviruses like group 2C MERS-CoV-related strains. It is notable that our chimeric spike vaccines and the SARS-CoV-2 furin KO, all of which lacked the two proline stabilizing mutations (S...

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   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).

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   Results from JetFighter: We did not find any issues relating to colormaps.

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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 10.1101/2021.03.11.434872 on bioRxiv

   Mar 12, 2021