Single Immunization with Recombinant ACAM2000 Vaccinia Viruses Expressing the Spike and the Nucleocapsid Proteins Protects Hamsters against SARS-CoV-2-Caused Clinical Disease
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Abstract
Continuous emergence of SARS-CoV-2 variants which cause breakthrough infection from the immunity induced by current spike protein-based COVID-19 vaccines highlights the need for new generations of vaccines that will induce long-lasting immunity against a wide range of the variants. To this end, we investigated the protective efficacy of the recombinant COVID-19 vaccine candidates based on a novel VACV ACAM2000 platform, in which an immunoregulatory gene, E3L, was deleted and both the SARS-CoV-2 spike (S) and nucleocapsid (N) antigens were expressed.
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SciScore for 10.1101/2021.12.02.470987: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: All the procedures were approved by the Animal Care Committee of the Canadian Science Centre for Human and Animal Health and followed the guidelines of the Canadian Council for Animal Care. Sex as a biological variable Animal experiment: Five to seven weeks old Golden Syrian hamsters were purchased from Charles River Laboratories (USA), and males and females were caged separately. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Expression of the S and N proteins were confirmed by Western blotting with a FLAG antibody. FLAGsuggested: NoneA 1:1000 dilution of Goat anti-hamster IgG … SciScore for 10.1101/2021.12.02.470987: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: All the procedures were approved by the Animal Care Committee of the Canadian Science Centre for Human and Animal Health and followed the guidelines of the Canadian Council for Animal Care. Sex as a biological variable Animal experiment: Five to seven weeks old Golden Syrian hamsters were purchased from Charles River Laboratories (USA), and males and females were caged separately. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Expression of the S and N proteins were confirmed by Western blotting with a FLAG antibody. FLAGsuggested: NoneA 1:1000 dilution of Goat anti-hamster IgG HRP conjugated antibody (ThermoFisher) was prepared in ChonBlock Secondary Antibody Dilution Buffer (Chondrex), 100µl was added and plates were incubated at room temperature for 1 hour. anti-hamster IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources A549/PKR-RNaseL-cells were used to select both ACAM2000ΔE3L and ACAM2000ΔE3LΔK3L viruses. A549/PKR-RNaseL-cellssuggested: NoneWestern blot analysis: HeLa and BHK21 Cell monolayers in a 12-well plate were infected with rACAM2000 viruses at an moi of 5 and the cell lysate was collected at 16 hours post infection with 200 µl of protein sample buffer. HeLasuggested: NoneBHK21suggested: NoneTriplicates of the serum and virus mixture were added to Vero cells in 96-well plates. Verosuggested: NoneRecombinant DNA Sentences Resources Carpenter using a pcDNA3.1/HEK293 eukaryotic expression system (unpublished). pcDNA3.1/HEK293suggested: NoneSoftware and Algorithms Sentences Resources Statistic analysis: Data were analyzed and plotted using GraphPad Prism 9 software. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: Thank you for sharing your data.
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 did not find any issues relating to the usage of bar graphs.
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.
Results from scite Reference Check: We found no unreliable references.
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