Prime-boost vaccinations with two serologically distinct chimpanzee adenovirus vectors expressing SARS-CoV-2 spike or nucleocapsid tested in a hamster COVID-19 model
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Abstract
Two serologically distinct replication-defective chimpanzee-origin adenovirus (Ad) vectors (AdC) called AdC6 and AdC7 expressing the spike (S) or nucleocapsid (N) proteins of an early SARS-CoV-2 isolate were tested individually or as a mixture in a hamster COVID-19 challenge model. The N protein, which was expressed as a fusion protein within herpes simplex virus glycoprotein D (gD) stimulated antibodies and CD8 + T cells. The S protein expressing AdC (AdC-S) vectors induced antibodies including those with neutralizing activity that in part cross-reacted with viral variants. Hamsters vaccinated with the AdC-S vectors were protected against serious disease and showed accelerated recovery upon SARS-CoV-2 challenge. Protection was enhanced if AdC-S vectors were given together with the AdC vaccines that expressed the gDN fusion protein (AdC-gDN). In contrast hamsters that just received the AdC-gDN vaccines showed only marginal lessening of symptoms compared to control animals. These results indicate that immune response to the N protein that is less variable that the S protein may potentiate and prolong protection achieved by the currently used genetic COVID-19 vaccines.
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SciScore for 10.1101/2022.03.17.484786: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Antibody Assays: ELISA for anti-S1 and anti-S2 antibodies: Sera from individual hamsters were tested for S-specific antibodies by ELISA on plates coated overnight with 100 µl of a mixture of S1 and S2 (Native Antigen Company, Kidlington, UK) each diluted to 1 µg/ml in bicarbonate buffer. anti-S1suggested: Noneanti-S2suggested: NoneAn anti-hamster IgG (H+L)-Alkaline Phosphatase antibody produced in goat (Sigma-Aldrich, St Louis, MO) diluted to 1:1000 in 3% BAS-PBS was added at 60 … SciScore for 10.1101/2022.03.17.484786: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Antibody Assays: ELISA for anti-S1 and anti-S2 antibodies: Sera from individual hamsters were tested for S-specific antibodies by ELISA on plates coated overnight with 100 µl of a mixture of S1 and S2 (Native Antigen Company, Kidlington, UK) each diluted to 1 µg/ml in bicarbonate buffer. anti-S1suggested: Noneanti-S2suggested: NoneAn anti-hamster IgG (H+L)-Alkaline Phosphatase antibody produced in goat (Sigma-Aldrich, St Louis, MO) diluted to 1:1000 in 3% BAS-PBS was added at 60 µl/well for 1 hour at room temperature. An anti-hamster IgGsuggested: Noneanti-hamster IgGsuggested: NoneH+L)-Alkaline Phosphatasesuggested: NoneELISA for RBD-binding antibodies: Sera were tested for inhibition of ACE binding to the RBD of the S protein by the Anti-SARS-CoV-2 Neutralizing Antibody Titer Serologic Assay Kit from Acro Biosystems (Newark, DE) following the manufacturer’s instructions. Anti-SARS-CoV-2suggested: NoneA positive standard with a known concentration provided by the kit was used to extrapolated anti-S antibody concentrations into µg per ml of serum. anti-Ssuggested: NoneThe VSV-S vector was diluted in medium containing 1 µg/ml of a mouse monoclonal antibody against VSV glycoprotein (sc-365019, Santa Crusz Biotechnology, Dallas, TX) and then incubated with the serum dilutions for 90 min at room temperature under gentle agitation starting with a serum dilution of 1 in 40. antibody against VSV glycoprotein (sc-365019, Santa Crusz Biotechnology, Dallas, TX)suggested: NoneA monoclonal antibody to N protein (Abbexa, MCA6373, lot# 156962) served as a positive control (data not shown). MCA6373suggested: NoneCells were then incubated with an anti-INF-γ-FITC antibody (clone XMG1.2 BioLegend, San Diego, CA) at 4°C for 30 min in the dark. anti-INF-γ-FITCsuggested: NoneExperimental Models: Cell Lines Sentences Resources Cell lines: HEK 293 cells were grown in Dulbecco’s Modified Eagles medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and antibiotics. HEK 293suggested: NoneChallenge virus: Challenge was conducted with the WT WAS stock, which was generated from seed stock (2019-nCoV/USA-WA1/2020), obtained from Biodefense and Emerging Infections Research (BEI) resources (Cat # NR-52281, Lot # 70036318) and expanded in Calu-3 cells at 37°C for 3 days. Calu-3suggested: NoneThe stock (Lot # 12152020-1235) titers in Vero TMPRSS2 cells are 2.34×109 Median Tissue Culture Infectious Dose (TCID50)/mL and 3.68 × 107 plaque forming units (PFU)/mL, and 6 × 105 PFU/mL in Vero76 cells. Vero TMPRSS2suggested: NoneVero76suggested: JCRB Cat# IFO50410, RRID:CVCL_0603)Generation of VSV-S vectors: VSV vectors pseudotyped with S of SARS-CoV-2 were generated in BHK-21/WI-2 cells using a the ΔG-GFP (G*ΔG-GFP) rVSV kit (Kerafast, Boston, MA, USA) and S sequences cloned into an expression plasmid under the control of the CMV promoter as described previously [51]. BHK-21/WI-2suggested: RRID:CVCL_HB78)A549A/T cells were plated into Terasaki plate wells as described above. A549A/Tsuggested: NoneExperimental Models: Organisms/Strains Sentences Resources A549-hACE-TMPRSS2 (A549A/T) cells (InvivoGen, cat no. a549-hace2tpsa) were maintained in DMEM with 10%FBS, 300µg/ml of hygromycin, 0.5 µg/ml of puromycin and antibiotics. A549-hACE-TMPRSS2 (A549A/Tsuggested: NoneRecombinant DNA Sentences Resources For the neutralization assay a dose of VSV-S vector that infected ∼20-40 cells/well was selected. VSV-Ssuggested: NoneSoftware and Algorithms Sentences Resources Post-acquisition analyses were performed with FlowJo (TreeStar, Ashland, OR). FlowJosuggested: (FlowJo, RRID:SCR_008520)The analyses were carried out by GraphPad Prism. GraphPad Prismsuggested: (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.
Results from scite Reference Check: We found no unreliable references.
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