Single-dose AAV-based vaccine induces a high level of neutralizing antibodies against SARS-CoV-2 in rhesus macaques
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SciScore for 10.1101/2021.05.19.444881: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: All procedures were conducted in accordance with the Principles for the Ethical Treatment of Animals approved by the Animal Care and Use Committee at the University of Science and Technology of China (202006220919000464981).
Euthanasia Agents: Hematoxylin-eosin (H&E) staining: Mice were anesthetized with sodium pentobarbital (40 mg/kg, intraperitoneal injection), then perfused with PBS and fixed in 4% paraformaldehyde (PFA).Sex as a biological variable Vaccine immunogenicity analysis: The C57BL/6J mice (8 weeks old; male and female; 20–25 g body weight) were randomly divided into five groups (5 mice per group). Randomization Vaccine immunogenicity analysis: The C57BL/6J mice (8 weeks old; … SciScore for 10.1101/2021.05.19.444881: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: All procedures were conducted in accordance with the Principles for the Ethical Treatment of Animals approved by the Animal Care and Use Committee at the University of Science and Technology of China (202006220919000464981).
Euthanasia Agents: Hematoxylin-eosin (H&E) staining: Mice were anesthetized with sodium pentobarbital (40 mg/kg, intraperitoneal injection), then perfused with PBS and fixed in 4% paraformaldehyde (PFA).Sex as a biological variable Vaccine immunogenicity analysis: The C57BL/6J mice (8 weeks old; male and female; 20–25 g body weight) were randomly divided into five groups (5 mice per group). Randomization Vaccine immunogenicity analysis: The C57BL/6J mice (8 weeks old; male and female; 20–25 g body weight) were randomly divided into five groups (5 mice per group). Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources The slices were then stained using 1:1◻000 anti-GFP antibody (Earthox, E002030-02) in blocking buffer overnight at 4 °C. anti-GFPsuggested: NoneSlices were washed with PBS three times (15 min each time) and incubated with secondary antibodies Alexa Fluor 488 donkey anti-mouse IgG (1: 1000, Thermo Scientific, A21202) for 2 h at room temperature. anti-mouse IgGsuggested: (Molecular Probes Cat# A-21202, RRID:AB_141607), anti-CD8 (BioLegend, 301007), and anti-CD20 (BioLegend, 302310) antibodies were added to the cells for staining for 30 min in the dark on ice. anti-CD8suggested: Noneanti-CD20suggested: (BioLegend Cat# 302310, RRID:AB_314258)), anti-TNF-α (BioLegend, 502930), and anti-IL-10 (BioLegend, 501420) antibodies and incubated on ice in the dark for 1 h. anti-TNF-αsuggested: Noneanti-IL-10suggested: (BioLegend Cat# 501420, RRID:AB_2125385)Experimental Models: Cell Lines Sentences Resources The HEK-293T cells were cultured with Dulbecco’s Modified Eagle Medium (DMEM, Gibco, 11965-092) and 10% fetal bovine serum (Gibco, 16000-044) at 37 °C. HEK-293Tsuggested: NoneThe virus-serum dilution mixtures and virus control were then inoculated into Vero E6 cell monolayers in 24-well plates for 1 h, before adding an overlay medium including 1.5% methylcellulose at 37 °C for 4–5 days to allow plaque development. Vero E6suggested: NoneExperimental Models: Organisms/Strains Sentences Resources Animal husbandry samples: C57BL/6 Mice and rhesus macaques (Macaca mulatta) were maintained at 25 °C on a 12 h: 12 h light:dark cycle in an animal room at the University of Science and Technology of China. C57BL/6suggested: NoneVaccine immunogenicity analysis: The C57BL/6J mice (8 weeks old; male and female; 20–25 g body weight) were randomly divided into five groups (5 mice per group). C57BL/6Jsuggested: RRID:IMSR_JAX:000664)Recombinant DNA Sentences Resources In brief, 10 μg of pHelper vector, 5 μg of AAV2/9 vector, and 5 μg of pITR vector (S2/S12/RBD/SRBD/GFP) were transfected into a 10-cm diameter dish with HEK-293T cells by Polyethylenimine (PEI) (PolyScience, Niles, USA). pHelpersuggested: NonepITRsuggested: NoneIn brief, target genes were inserted into the pTT5 vector, which contains a IFNA1 signal peptide at the N-terminus and a TEV enzyme site connected to the human IgG1 Fc at the C-terminus. pTT5suggested: RRID:Addgene_52326)Software and Algorithms Sentences Resources The data were fitted by Prism v6 to calculate the Tm values. Prismsuggested: (PRISM, RRID:SCR_005375)FlowJo v10 software was used for data analysis. FlowJosuggested: (FlowJo, RRID:SCR_008520)Quantification graphs were analyzed using GraphPad Prism v6 (GraphPad Software). GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)GraphPadsuggested: (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 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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