Intranasal pediatric parainfluenza virus-vectored SARS-CoV-2 vaccine is protective in monkeys
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SciScore for 10.1101/2022.05.21.492923: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: Immunization and challenge of rhesus macaques: All animal studies were approved by the NIAID Animal Care and Use Committee. Sex as a biological variable Eight juvenile to young adult male Indian-origin rhesus macaques (Macaca mulatta), confirmed to be seronegative for HPIV3 and SARS-CoV-2, were immunized intranasally (0.5 ml per nostril) and intratracheally (1 ml) with a total does of 6.3 log10 plaque-forming units (PFU) of B/HPIV3/S-6P or the empty vector control B/HPIV3. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Infected monolayers were overlaid with culture medium … SciScore for 10.1101/2022.05.21.492923: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: Immunization and challenge of rhesus macaques: All animal studies were approved by the NIAID Animal Care and Use Committee. Sex as a biological variable Eight juvenile to young adult male Indian-origin rhesus macaques (Macaca mulatta), confirmed to be seronegative for HPIV3 and SARS-CoV-2, were immunized intranasally (0.5 ml per nostril) and intratracheally (1 ml) with a total does of 6.3 log10 plaque-forming units (PFU) of B/HPIV3/S-6P or the empty vector control B/HPIV3. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Infected monolayers were overlaid with culture medium containing 0.8% methylcellulose, and incubated at 32°C for 6 days, fixed with 80% methanol, and immunostained with a rabbit hyperimmune serum raised against purified HPIV3 virions to detect B/HPIV3 antigens, and a goat hyperimmune serum to the secreted SARS-CoV-2 S to detect co-expression of the S protein, followed by infrared-dye conjugated donkey anti-rabbit IRDye680 IgG and donkey anti-goat IRDye800 IgG secondary antibodies (LiCor). anti-rabbit IRDye680 IgGsuggested: Noneanti-goatsuggested: NoneDissociation-enhanced lanthanide fluorescent (DELFIA) time resolved fluorescence (TRF) immunoassay, ELISA and live HPIV3 and SARS-CoV-2 neutralization assay: Levels of anti-SARS-CoV-2 S antibodies elicited by B/HPIV3/S-6P were determined by DELFIA-TRF (Perkin Elmer) from NW or BAL following the supplier’s protocol and from serum samples by ELISA (Liu et al., 2021) using the recombinantly-expressed secreted version of S-2P (Wrapp et al., 2020), or a fragment (aa 328-531) containing the receptor binding domain (RBD) of the SARS-CoV-2 S protein (Walls et al., 2020). anti-SARS-CoV-2 Ssuggested: NoneThe secondary antibodies used in both assays were goat anti-monkey IgG(H+L) horseradish peroxidase (HRP) (Thermo Fisher, Cat #PA1-84631) anti-monkey IgG(H+Lsuggested: NoneThe antibodies used for extracellular and intracellular staining were: CD69 (FITC, clone FN50, Biolegend) CD69suggested: NoneExperimental Models: Cell Lines Sentences Resources African green monkey kidney Vero (ATCC CCL-81) and Vero E6 (ATCC CRL-1586) cells were cultured in Dulbecco’s MEM with GlutaMAX (Thermo Fisher Scientific) with 5% FBS and 1% L-glutamine. Vero E6suggested: NoneThe B/HPIV3/S-6P cDNA was used to transfect BHK21 cells (clone BSR T7/5, stably expressing T7 RNA polymerase (Buchholz et al., 1999)), together with helper plasmids encoding the N, P and L proteins (Buchholz et al., 2004; Liu et al., 2021), to produce the B/HPIV3/S-6P recombinant virus. BHK21suggested: NoneBriefly, Vero cell monolayers in 24-well plates were infected in duplicate with 10-fold serially diluted samples. Verosuggested: NoneRecombinant DNA Sentences Resources , luciferase reporter (pHR’ CMV Luc), lentivirus backbone (pCMV ΔR8.2), and human transmembrane protease serine 2 (TMPRSS2) at a ratio of 1:20:20:0.3 into HEK293T/17 cells (ATCC) with transfection reagent LiFect293™. pCMV ΔR8.2suggested: NoneStandard curves were generated using serially diluted pcDNA3.1 plasmids encoding gN, gE, or sgE sequences. pcDNA3.1suggested: RRID:Addgene_79663)Software and Algorithms Sentences Resources IC50 titers were determined using a log (agonist) vs. normalized response (variable slope) nonlinear function in Prism v8 (GraphPad). GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)Data were analyzed using FlowJo version 10. FlowJosuggested: (FlowJo, RRID:SCR_008520)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: We found the following clinical trial numbers in your paper:
Identifier Status Title NCT04816643 Recruiting A Phase 1/2/3 Study to Evaluate the Safety, Tolerability, an… NCT00686075 Completed A Study to Evaluate the Safety, Tolerability, Immunogenicity… 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.
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Results from scite Reference Check: We found no unreliable references.
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