Rapid protection from COVID-19 in nonhuman primates vaccinated intramuscularly but not intranasally with a single dose of a recombinant vaccine

Article activity feed

1. 

   ScreenIT

   Jan 20, 2021

   SciScore for 10.1101/2021.01.19.426885: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: All procedures followed standard operating procedures (SOPs) approved by the RML Institutional Biosafety Committee (
   Randomization	The NHPs were randomly selected for two vaccine groups (n=6) and one control group (n=4).
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Animal study: Sixteen female rhesus macaques (3.5-10 years of age; 4.5-10kg, Indian-origin) were used in this study.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   After horse-radish peroxidase (HRP)-labeled secondary antibody staining using either anti-mouse IgG (1:10,000) or anti-rabbit IgG (1:5000) (Jackson ImmunoResearch), the …

   SciScore for 10.1101/2021.01.19.426885: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: All procedures followed standard operating procedures (SOPs) approved by the RML Institutional Biosafety Committee (
   Randomization	The NHPs were randomly selected for two vaccine groups (n=6) and one control group (n=4).
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Animal study: Sixteen female rhesus macaques (3.5-10 years of age; 4.5-10kg, Indian-origin) were used in this study.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   After horse-radish peroxidase (HRP)-labeled secondary antibody staining using either anti-mouse IgG (1:10,000) or anti-rabbit IgG (1:5000) (Jackson ImmunoResearch), the blots were imaged using the SuperSignal West Pico chemiluminescent substrate (Thermo Fisher Scientific) and an iBright™ CL1500 Imaging System (Thermo Fisher Scientific)	anti-mouse IgG suggested: (LSBio (LifeSpan Cat# LS-C69682-5000, RRID:AB_1653096) anti-rabbit IgG suggested: None
   After 3 washes with PBST, the bound antibodies were labeled using 50 μl of 1:2,500 horse-radish peroxidase (HRP)-labeled anti-monkey IgG (H+L) (SeraCare Life Sciences) diluted in 1% skim milk in PBST.	anti-monkey IgG suggested: None
   Specific anti-CoV immunoreactivity was detected using an in-house SARS-CoV-2 nucleocapsid protein (U864YFA140-4/CB2093) rabbit antibody (Genscript) at a 1:1000 dilution.	SARS-CoV-2 nucleocapsid protein ( U864YFA140-4/CB2093 ) suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Cells and Viruses: Huh7 and VeroE6 cells were grown at 37°C and 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) (Sigma-Aldrich, St. Louis, MO) containing 10% fetal bovine serum (FBS) (Wisent Inc., St. Bruno, Canada), 2 mM L-glutamine (Thermo Fisher Scientific, Waltham, MA), 50 U/mL penicillin (Thermo Fisher Scientific), and 50 μg/mL streptomycin (Thermo Fisher Scientific).	Huh7 suggested: None
   The replication-competent recombinant VSV was recovered in BHK-T7 cells as described previously (26).	BHK-T7 suggested: None
   Finally, media was removed from cells and the mixture was added to VeroE6 cells and incubated at 37°C and 5% CO2 for 6 days.	VeroE6 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
   Software and Algorithms
   Sentences	Resources
   Sample acquisition was performed on a Cytoflex-S (Beckman Coulter) and data analyzed in FlowJo V10 (TreeStar).	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   RNA-Seq reads were demultiplexed, quality-filtered and trimmed using Trim Galore (average Phred score cut-off of 30, minimum length of 50 bp).	Trim Galore suggested: (Trim Galore, RRID:SCR_011847)
   FastQC was used to generate quality reports.	FastQC suggested: (FastQC, RRID:SCR_014583)
   Hisat2 was used to align reads to the reference genome Macaca mulatta (Macaca_mulatta.Mmul_8.0.1.dna.toplevel.fa) and the Macaca_mulatta.Mmul_8.0.1.97.gtf was used for annotation.	Hisat2 suggested: (HISAT2, RRID:SCR_015530)
   Raw expression values (gene-level read counts) were generated using the summarizeOverlaps function and normalized (read per kilobase of transcript per million mapped reads, rpkm) using the edgeR package.	edgeR suggested: (edgeR, RRID:SCR_012802)
   Functional enrichment of DEGs was performed using Metascape to identify relevant Gene Ontology (GO) biological process terms and KEGG pathways.	Metascape suggested: (Metascape, RRID:SCR_016620) KEGG suggested: (KEGG, RRID:SCR_012773)
   In silico flow cytometry was performed using ImmQuant with the IRIS database.	IRIS suggested: None
   , Venn diagrams and violin plots were generated using R packages ggplot2 and VennDiagrams.	ggplot2 suggested: (ggplot2, RRID:SCR_014601)
   GO network plots were generated in Cytoscape (Version 3.5.1).	Cytoscape suggested: (Cytoscape, RRID:SCR_003032)
   Graphs were generated using GraphPad Prism software (version 8).	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)
   Statistical analyses: All statistical analysis was performed in Prism 8 (GraphPad).	GraphPad 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: 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.

   ---

   Read the original source
2. 

   Version published to 10.1101/2021.01.19.426885 on bioRxiv

   Jan 19, 2021