An AAV-ie based Vaccine effectively protects against SARS-CoV-2 and Circulating Variants

Article activity feed

1. 

   ScreenIT

   May 23, 2021

   SciScore for 10.1101/2021.05.19.444889: (What is this?)

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

   Table 1: Rigor

   Ethics

   Euthanasia Agents: After 2 weeks, mice were anesthetized with sodium pentobarbital (40 mg/kg, i.p.) with no avoidance response to foot pinch. IACUC: Following imaging was performed using confocal microscope (Nikon) Mouse vaccination: The mouse study was performed under the guidance of Institutional Animal Care and Use Committee (IACUC) of Shanghaitech University, China. Male BALB/c mice aging 6 – 8 weeks were used in this study.

   Sex as a biological variable

   Following imaging was performed using confocal microscope (Nikon) Mouse vaccination: The mouse study was performed under the guidance of Institutional Animal Care and Use Committee (IACUC) of Shanghaitech University, China. Male BALB/c mice …

   SciScore for 10.1101/2021.05.19.444889: (What is this?)

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

   Table 1: Rigor

   Ethics	Euthanasia Agents: After 2 weeks, mice were anesthetized with sodium pentobarbital (40 mg/kg, i.p.) with no avoidance response to foot pinch. IACUC: Following imaging was performed using confocal microscope (Nikon) Mouse vaccination: The mouse study was performed under the guidance of Institutional Animal Care and Use Committee (IACUC) of Shanghaitech University, China. Male BALB/c mice aging 6 – 8 weeks were used in this study.
   Sex as a biological variable	Following imaging was performed using confocal microscope (Nikon) Mouse vaccination: The mouse study was performed under the guidance of Institutional Animal Care and Use Committee (IACUC) of Shanghaitech University, China. Male BALB/c mice aging 6 – 8 weeks were used in this study.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Then, the NC membrane was incubated for 1 hour in 5% milk PBST with a 1:1,000 dilution of mouse anti-His tag antibody (Proteintech, Cat# 66005-1-lg).	anti-His tag suggested: None
   The slices were then stained using rabbit anti-GFP antibody (Proteintech, Cat# 50430-2-AP) in 0.1% Triton X-100 and 1% serum in PBS overnight at 4 °C.	anti-GFP suggested: (Proteintech Cat# 50430-2-AP, RRID:AB_11042881)
   After washing with PBS, sections were incubated with the Alexa-488 conjugated donkey anti-rabbit antibody (Thermo Fisher Scientific, Cat# A21206) (1:1000 dilution) for 2 h at room temperature.	anti-rabbit suggested: (Thermo Fisher Scientific Cat# A-21206, RRID:AB_2535792)
   After washes with PBS, HRP-conjugated secondary antibodies (HRP conjugated goat anti-mouse IgG: Proteintech, Cat# SA00001-1; HRP conjugated goat anti-mouse IgG2a: Proteintech, Cat# SA00012-2; HRP conjugated goat anti-mouse IgG1: Proteintech, Cat# SA00012-1; HRP-conjugated goat anti-monkey IgG: Southern Biotech, Cat# 4700-05) were added and incubated for 1 h.	anti-mouse IgG suggested: (Proteintech Cat# SA00012-2, RRID:AB_2890965) anti-mouse IgG2a suggested: (Gen-Probe Cat# 855.470.005, RRID:AB_10698621) anti-mouse IgG1 suggested: (Gen-Probe Cat# 855.470.005, RRID:AB_10698621) anti-monkey IgG suggested: (SouthernBiotech Cat# 4700-05, RRID:AB_2796069)
   Experimental Models: Cell Lines
   Sentences	Resources
   The plasmids were transfected into HEK Expi293F cells (Thermo Fisher Scientific, Cat# A14527) using polyethylenimine (PEI) method.	HEK Expi293F suggested: None
   Pseudo-virus neutralization assay: To prepare SARS-CoV-2 and related mutated pseudo-viruses, HEK-293T cells were co-transfected with pcDNA3.1-SARS-CoV-2-S or related mutants and pNL4-3.luc.	HEK-293T suggested: None
   To test the neutralizing activity of immune sera against pseudo-virus, HEK293T cells stably transfected with hACE2 were seeded in 96-well culture plates at a density of 5,000 cells per well.	HEK293T suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Following imaging was performed using confocal microscope (Nikon) Mouse vaccination: The mouse study was performed under the guidance of Institutional Animal Care and Use Committee (IACUC) of Shanghaitech University, China. Male BALB/c mice aging 6 – 8 weeks were used in this study.	BALB/c suggested: RRID:IMSR_ORNL:BALB/cRl)
   Recombinant DNA
   Sentences	Resources
   The coding sequences of S-trimer ECD (residues 1-1208) or RBD were cloned into pTT5 vector with a N-terminal IL-2 signal peptide and a C-terminal 6 × His tag.	pTT5 suggested: RRID:Addgene_52326)
   Pseudo-virus neutralization assay: To prepare SARS-CoV-2 and related mutated pseudo-viruses, HEK-293T cells were co-transfected with pcDNA3.1-SARS-CoV-2-S or related mutants and pNL4-3.luc.	pcDNA3.1-SARS-CoV-2-S suggested: None pNL4-3 suggested: None
   Software and Algorithms
   Sentences	Resources
   Statistics: Antibody titers of ELISA assay, EC50 value of pseudo-virus neutralization assay were determined using non-linear regression analysis using Graphpad PRISM.	Graphpad PRISM suggested: (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: Please consider improving the rainbow (“jet”) colormap(s) used on pages 10 and 21. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

   ---

   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.

   ---

   Read the original source
2. 

   Version published to 10.1101/2021.05.19.444889v1 on bioRxiv

   May 20, 2021