Boosting of SARS-CoV-2 immunity in nonhuman primates using an oral rhabdoviral vaccine

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1. Version published to 10.1016/j.vaccine.2021.12.063

   Apr 1, 2022
2. 

   ScreenIT

   Oct 22, 2021

   SciScore for 10.1101/2021.10.16.464660: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: Nonhuman primate study: Ethics statement: This study was conducted as approved by the University of Minnesota Institutional Animal Care and Use Committee.
   Sex as a biological variable	Animals: Twenty-two adult male and female adult Mauritian origin cynomolgus macaques (Macaca fascicularis) were allocated for this study.
   Randomization	In experiment 1, eighteen adult males and females with the median age of 3.7 years (range 3.4 to 6.7 years) and median body weight of 4.8 kg (3.2 to 6.8kg) were stratified by sex and randomly assigned to control and vaccine groups (Fig. 2).
   Blinding	These tissues were evaluated histologically for signs of pathology by a board-certified veterinary pathologist …

   SciScore for 10.1101/2021.10.16.464660: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: Nonhuman primate study: Ethics statement: This study was conducted as approved by the University of Minnesota Institutional Animal Care and Use Committee.
   Sex as a biological variable	Animals: Twenty-two adult male and female adult Mauritian origin cynomolgus macaques (Macaca fascicularis) were allocated for this study.
   Randomization	In experiment 1, eighteen adult males and females with the median age of 3.7 years (range 3.4 to 6.7 years) and median body weight of 4.8 kg (3.2 to 6.8kg) were stratified by sex and randomly assigned to control and vaccine groups (Fig. 2).
   Blinding	These tissues were evaluated histologically for signs of pathology by a board-certified veterinary pathologist blinded to study group allocations.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Correct incorporation of VSV G, N and M proteins and S glycoprotein in virions or virus infected cells was analyzed by Western blotting using a mouse α-SARS-CoV-2 Spike (1:1000, GeneTex #GTX632604), rabbit polyclonal α-VSV-G antibody (1:20,000, Abcam #ab83196), and rabbit polyclonal α-VSV antiserum (1:8000, Imanis #REA-005).	α-SARS-CoV-2 suggested: None α-VSV-G suggested: None
   Secondary antibodies were goat α-mouse IgG-HRP (1:30,000, Prometheus #20-304) and goat α-rabbit IgG-HRP (1:30,000, Prometheus #20-303).	α-mouse IgG-HRP suggested: None α-rabbit IgG-HRP suggested: None
   Plates were washed three times with PBS with 0.05% Tween 20 and then incubated for 45 minutes at RT with either horse radish peroxidase (HRP)-conjugated anti-NHP IgG (1:10,000, Cat.# PA1-84631, Invitrogen) or HRP-conjugated anti-NHP IgA (1:5,000, Cat.# 5220-0332, SeraCare) secondary antibodies diluted in sample buffer.	anti-NHP IgG suggested: None anti-NHP IgA suggested: None
   Pooled seronegative NHP serum was used as a negative control, and pooled seronegative NHP serum spiked with monoclonal anti-SARS-CoV-2 spike antibody was used as a positive control.	anti-SARS-CoV-2 suggested: None
   VSV-G Antibody Neutralization Assays of NHP Samples: NHP serum samples and positive control rabbit α-VSV antiserum (Imanis #REA-005) were heat inactivated at 56°C for 30 min.	α-VSV suggested: None
   Plates were washed three times with PBS with 0.05% Tween 20 and then incubated for 1h at RT with horse radish peroxidase (HRP)-conjugated anti-mouse IgG (1:5,000, Cat.#62-6520, ThermoFisher Scientific), IgG1 (1:5,000, Cat.# 115-035-205, Jackson ImmunoResearch) or IgG2a (1:5,000, Cat.# 115-035-206, Jackson ImmunoResearch) secondary antibody.	anti-mouse IgG suggested: (Jackson ImmunoResearch Labs Cat# 115-035-205, RRID:AB_2338513) IgG1 suggested: None IgG2a suggested: None
   , low-fluorescent PVDF membrane (Millipore, Bedford, MA, USA) plates were coated with IFNγ and IL-4 capture antibodies overnight.	IL-4 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   The viruses were amplified and propagated in Vero cells.	Vero suggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)
   IMMUNO-COV™ Neutralization Assays of NHP Samples: Vero-ACE2 cells were seeded at 104 cells/well in 96-well black-walled plates with clear bottoms 16-24 h before being used for neutralization assays.	Vero-ACE2 suggested: None
   After a 1 hour incubation at 37°C, 2×104 BHK-21 cells were overlaid onto the virus mixes.	BHK-21 suggested: ATCC Cat# CRL-6281, RRID:CVCL_1914)
   And the mixture was added in quadruplicates to hACE-2-expressing HEK293T cells overnight, followed by media replenishment.	HEK293T suggested: CCLV Cat# CCLV-RIE 1018, RRID:CVCL_0063)
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Female and male Ifnartm-CD46Ge mice [31] received intra-peritoneally 5×105 TCID50 virus particles or 5μg of SARS-CoV-2 spike protein (Cat.# V0589-V08B, Sino Biological) adjuvanted with aluminum (Cat.#vac-alu-250,InvivoGen) on day 0 and 21.	Ifnartm-CD46Ge suggested: None
   Recombinant DNA
   Sentences	Resources
   Materials and Methods: Virus generation and characterization: Full-length human codon-optimized SARS-CoV-2 Spike (S) glycoprotein (NC_045512.2) in pUC57 was obtained from GenScript (MC_0101081).	pUC57 suggested: RRID:Addgene_40306)
   To generate the viral genome, the amplified PCR product containing S-Δ19CT was cloned into pVSV in place of VSV-G using the MluI and NheI restriction sites (Fig.	pVSV suggested: None
   In brief, HEK293T cells seeded overnight were transfected with HDM-Hgpm2, HDM-tat1b, pRC-CMV-Rev1b, pHAGE-CMV-Luc2-IRES-ZsGreen-W and a codon-optimized SARS-CoV-2 spike encoding the D614G amino acid change.	pRC-CMV-Rev1b suggested: RRID:Addgene_164443) pHAGE-CMV-Luc2-IRES-ZsGreen-W suggested: RRID:Addgene_164432)
   Software and Algorithms
   Sentences	Resources
   The half maximal inhibitory concentration (IC50) was determined by non-linear regression using GraphPad Prism version 8.4.2 for macOS (GraphPad Software, San Diego, CA, USA).	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798) GraphPad 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: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

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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.
   • No funding statement was detected.
   • No protocol registration statement was detected.

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   Results from scite Reference Check: We found no unreliable references.

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   Version published to 10.1101/2021.10.16.464660 on bioRxiv

   Oct 18, 2021