mRNA-1273 or mRNA-Omicron boost in vaccinated macaques elicits comparable B cell expansion, neutralizing antibodies and protection against Omicron

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   ScreenIT

   Feb 8, 2022

   SciScore for 10.1101/2022.02.03.479037: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: Rhesus macaque model and immunizations: All experiments conducted according to NIH regulations and standards on the humane care and use of laboratory animals as well as the Animal Care and Use Committees of the NIH Vaccine Research Center and BIOQUAL, Inc. (Rockville, Maryland).
   Sex as a biological variable	not detected.
   Randomization	not detected.
   Blinding	Subgenomic RNA quantification: sgRNA was isolated and quantified by researchers blinded to vaccine history as previously described (Corbett et al., 2021c), except for the use of a new probe noted below.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Briefly, total IgG …

   SciScore for 10.1101/2022.02.03.479037: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: Rhesus macaque model and immunizations: All experiments conducted according to NIH regulations and standards on the humane care and use of laboratory animals as well as the Animal Care and Use Committees of the NIH Vaccine Research Center and BIOQUAL, Inc. (Rockville, Maryland).
   Sex as a biological variable	not detected.
   Randomization	not detected.
   Blinding	Subgenomic RNA quantification: sgRNA was isolated and quantified by researchers blinded to vaccine history as previously described (Corbett et al., 2021c), except for the use of a new probe noted below.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Briefly, total IgG antigen-specific antibodies to variant SARS-CoV-2 S- and RBD-derived antigens were determined in a multiplex serology assay by MSD V-Plex SARS-CoV-2 Panel 23 for S and MSD V-Plex SARS-CoV-2 Panel 22 for RBD) according to manufacturer’s instructions, except 25μl of sample and detection antibody were used per well.	total IgG suggested: None
   Cells were incubated with an anti-SARS-CoV S primary antibody directly conjugated to Alexaflour-647 (CR3022-AF647) overnight at 4°C.	anti-SARS-CoV S suggested: None
   Briefly, ELISA against S-2P was performed in the absence or presence of sodium thiocyanate (NaSCN) and developed with HRP-conjugated goat anti-monkey IgG (H+L) secondary antibody (Invitrogen) and SureBlue 3,3′,5,5′-tetramethylbenzidine (TMB) microwell peroxidase substrate (1-Component; SeraCare) and quenched with 1 N H2SO4.	anti-monkey IgG suggested: None
   , anti-histidine antibody was immobilized on Series S Sensor Chip CM5 (Cytiva) allowing for the capture of his-tagged SARS-CoV-2 S-2P on active sensor surface.	anti-histidine suggested: None
   Human IgG monoclonal antibodies (mAbs) used for these analyses include: RBD-specific mAbs B1-182, A19-46.1, A20-29.1, A19-61.1, S309, A23-97.1 and A23-80.1.	A19-61.1 suggested: (ABclonal Cat# A19611, RRID:AB_2862699) A23-80.1 suggested: None
   The following monoclonal antibodies were used: CD3 APC-Cy7 (clone SP34-2, BD Biosciences), CD4 PE-Cy5.5 (clone S3.5, Invitrogen), CD8 BV570 (clone RPA-T8, BioLegend)	CD4 suggested: (BioLegend Cat# 100541, RRID:AB_10897943) CD8 suggested: None
   Briefly, seven to nine days following Omicron challenge animals were euthanized and lung tissue was processed and stained with hematoxylin and eosin for pathological analysis or with a rabbit polyclonal anti-SARS-CoV-2 anti-nucleocapsid antibody (GeneTex, GTX135357) at a dilution of 1:2000 for IHC.	anti-SARS-CoV-2 anti-nucleocapsid antibody (GeneTex, GTX135357 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Method details: Cells and viruses: VeroE6-TMPRSS2 cells were generated at the Vaccine Research Center, NIH, Bethesda, MD.	VeroE6-TMPRSS2 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
   Lentiviral pseudovirus neutralization: Neutralizing antibodies in serum or plasma were measured in a validated pseudovirus-based assay as a function of reductions in luciferase reporter gene expression after a single round of infection with SARS-CoV-2 spike-pseudotyped viruses in 293T/ACE2 cells (293T cell line stably overexpressing the human ACE2 cell surface receptor protein, obtained from Drs.	293T/ACE2 suggested: RRID:CVCL_YZ65) 293T suggested: KCB Cat# KCB 200744YJ, RRID:CVCL_0063)
   SARS-CoV-2 S-pseudotyped virus was prepared by transfection in 293T/17 cells (human embryonic kidney cells in origin; obtained from American Type Culture Collection, #CRL-11268) using a lentivirus backbone vector, a spike-expression plasmid encoding S protein from Wuhan-Hu-1 strain (GenBank no. NC_045512) with a p.Asp614Gly mutation, a TMPRSS2 expression plasmid and a firefly Luc reporter plasmid.	293T/17 suggested: ATCC Cat# CRL-11268, RRID:CVCL_1926)
   Briefly, Vero-TMPRSS2 cells were plated and incubated overnight.	Vero-TMPRSS2 suggested: JCRB Cat# JCRB1818, RRID:CVCL_YQ48)
   Software and Algorithms
   Sentences	Resources
   The FRNT-50 titers were interpolated using a 4-parameter nonlinear regression in GraphPad Prism 9.2.0.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)
   Samples were acquired on an BD FACSymphony cytometer and analyzed using FlowJo version 10.7.2 (BD, Ashland, OR)	FlowJo suggested: (FlowJo, RRID:SCR_008520)

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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: We detected the following sentences addressing limitations in the study:

   Limitations of the study: There are several limitations to this study. First, NHP models may not fully recapitulate clinical data in humans regarding the extent of viral replication necessary for the enhanced transmission of Omicron compared to prior variants. Even if a significant component of Omicron’s growth advantage is due to immune escape, the role of viral replication kinetics cannot be ruled out. Here, viral titers were low in the lungs and low to undetectable in the upper airway. Second, neutralizing antibody titers in NHP are 5-to 10-fold greater than in humans that received the same dose and regimen of mRNA-1273 with a boost (Edara et al., 2021a; Pajon et al., 2022). Third, a second dose of mRNA-Omicron may elicit a population of B cells specific only for Omicron. Finally, since we sought to compare two different mRNA boosts, we did not have an unboosted group to determine whether the boost enhanced protection. As all the boosted NHP were completely protected in the lungs, we were unable to determine an immune threshold for protection.

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

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   Version published to 10.1101/2022.02.03.479037v1 on bioRxiv

   Feb 4, 2022