Single-dose intranasal administration of AdCOVID elicits systemic and mucosal immunity against SARS-CoV-2 in mice

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   ScreenIT

   Oct 15, 2020

   SciScore for 10.1101/2020.10.10.331348: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: Studies performed at UAB were performed under IACUC Approval Number 21203.
   Randomization	Vaccination: Inbred female C57BL/6J (The Jackson Laboratory) and outbred CD-1 (Charles River Laboratories) mice of at least 6 weeks of age were randomly allocated into vaccination groups.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Vaccination: Inbred female C57BL/6J (The Jackson Laboratory) and outbred CD-1 (Charles River Laboratories) mice of at least 6 weeks of age were randomly allocated into vaccination groups.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   After removal of the mouse …

   SciScore for 10.1101/2020.10.10.331348: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: Studies performed at UAB were performed under IACUC Approval Number 21203.
   Randomization	Vaccination: Inbred female C57BL/6J (The Jackson Laboratory) and outbred CD-1 (Charles River Laboratories) mice of at least 6 weeks of age were randomly allocated into vaccination groups.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Vaccination: Inbred female C57BL/6J (The Jackson Laboratory) and outbred CD-1 (Charles River Laboratories) mice of at least 6 weeks of age were randomly allocated into vaccination groups.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   After removal of the mouse anti-adenovirus-5 hexon antibody and additional phosphate-buffered saline washes,	anti-adenovirus-5 suggested: None
   HRP-conjugated rat anti-mouse antibody was added to each well and incubated at least 60 minutes at 37°C.	anti-mouse suggested: None
   The loading of recombinant SARS2-CoV-2 spike onto the beads was evaluated by staining 1E+05 beads with dilutions ranging from 1μg to 2ng/ml of the recombinant anti-SARS spike antibody CR3022 and visualized with an anti-human IgG secondary antibody.	anti-SARS suggested: None anti-human IgG suggested: None
   Briefly, 0.2 mg of goat polyclonal anti-mouse IgG (southern Biotech cat# 1013-01), anti-IgM (cat#1 022-01), and anti-IgA (cat# 1040-01) antibodies in PBS were mixed with 5E+07 fluorescent microparticles each with a unique fluorescent intensity in the far red channels (Spherotech 3.6um cat# CPAK-3567-4K, peaks 1-3) resuspended in 0.1 M MES buffer pH 5.0.	anti-mouse IgG suggested: (SouthernBiotech Cat# 1013-01, RRID:AB_2794188) anti-IgM suggested: None anti-IgA suggested: None
   For primary antibody detection, a SARS-CoV-2 spike/RBD antibody (Rabbit, Polyclonal, SinoBiologicals Ct No. 40592-T62) was added to 5% Blotto and incubated on the monolayers overnight.	spike/RBD suggested: None
   Samples were incubated for 10 minutes at 4°C in the dark with Fc-Block (clone 24G2, 10 μg/ml), washed with 200 µl staining media (SME) and then stained with myeloid, B cell or BAL antibody panels.	BAL suggested: None
   Cells were then incubated on ice with a combination of fluorescent dye-labelled antibodies including anti-CD4-V500 (clone GK1.5; 1:200 dilution), anti-CD8α-APC-Fire750 (clone 53-6.7; 1:200 dilution), anti-CD11a/CD18-Pacific Blue (H155-78; 1:200 dilution), anti-CD103-PE (M290; 1:200 dilution), anti-CD69-FITC (H1-2F3; 1:200 dilution), anti-Ly6G-PerCP-Cy5.5 (clone 1A8; 1:200 dilution), anti-CD64-PerCP-Cy5.5 (clone X54-5/7.1; 1:200 dilution), anti-B220/CD45R-PerCP (clone RA3-6B2; 1:200 dilution), and Red LIVE/DEAD (1:1000 dilution).	anti-CD4-V500 suggested: None anti-CD8α-APC-Fire750 suggested: None anti-CD11a/CD18-Pacific suggested: None anti-CD103-PE suggested: None M290 suggested: None anti-CD69-FITC suggested: (Sigma-Aldrich Cat# SAB4700221, RRID:AB_10898330) anti-Ly6G-PerCP-Cy5.5 suggested: None anti-CD64-PerCP-Cy5.5 suggested: None anti-B220/CD45R-PerCP suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Adenovirus vaccine titer measurement: 293 HEK cells were seeded in a 96-well plate one day before Ad vector infection.	HEK suggested: None
   Vero E6 cells were grown on 96-well plates to confluence.	Vero E6 suggested: None
   After incubation, the sera:virus mixtures were added to the wells (100µL), and infection allowed to proceed for 1 hour on the Vero cells at 35°C.	Vero suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   CD-1).	CD-1 suggested: RRID:MGI:2686808)
   Dose levels used included high-dose (3.35E+08 ifu), mid-dose (6E+07 ifu) or low-dose (6E+06 ifu) of vaccine except for the S1 vector tested C57BL/6J mice were the high dose vaccine was 6E+08 ifu.	C57BL/6J suggested: None
   Software and Algorithms
   Sentences	Resources
   Ethics statement and Mice: Mice used in these studies were obtained from the Jackson Laboratory (C56BL/6J) or Charles River Laboratories (	Charles River Laboratories suggested: (Charles River Laboratories, RRID:SCR_003792)
   Following acquisition, the resulting FCS files were analyzed in flowJo (treestar).	flowJo suggested: (FlowJo, RRID:SCR_008520)
   Plates were washed 5X with PBS, and further incubated with a goat anti-rabbit IgG conjugated to horseradish peroxidase (Boster Biological Technology Co., #BA1054-0.5) in 5% Blotto for 1 hr.	Boster Biological suggested: None
   Quantification and statistical analysis: Statistical significance was assigned when P values were < 0.05 using Prism Version 8.4.2 (GraphPad).	Prism suggested: (PRISM, RRID:SCR_005375) 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 did not find any issues relating to the usage of bar graphs.

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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.
   • 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.

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2. 

   Version published to 10.1101/2020.10.10.331348v1 on bioRxiv

   Oct 11, 2020