Intranasal Immunization with a Vaccinia Virus Vaccine Vector Expressing Pre-Fusion Stabilized SARS-CoV-2 Spike Fully Protected Mice against Lethal Challenge with the Heavily Mutated Mouse-Adapted SARS2-N501YMA30 Strain of SARS-CoV-2
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Abstract
The Omicron SARS-CoV-2 variant has been designated as a variant of concern because its spike protein is heavily mutated. In particular, the Omicron spike is mutated at five positions (K417, N440, E484, Q493, and N501) that have been associated with escape from neutralizing antibodies induced by either infection with or immunization against the early Washington strain of SARS-CoV-2. The mouse-adapted strain of SARS-CoV-2, SARS2-N501YMA30, contains a spike that is also heavily mutated, with mutations at four of the five positions in the Omicron spike associated with neutralizing antibody escape (K417, E484, Q493, and N501). In this manuscript, we show that intranasal immunization with a pre-fusion stabilized Washington strain spike, expressed from a highly attenuated, replication-competent vaccinia virus construct, NYVAC-KC, fully protected mice against symptoms and death from SARS2-N501YMA30. Similarly, immunization by scarification on the skin fully protected against death, but not from mild disease. This data demonstrates that the Washington strain spike, when expressed from a highly attenuated, replication-competent poxvirus—administered without parenteral injection—can fully protect against the heavily mutated mouse-adapted SARS2-N501YMA30.
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SciScore for 10.1101/2021.12.06.471483: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: Symptoms were scored in a blinded manner for ruffled fur, hunching and activity, and scored from 0-3 (0 normal, 3 severe) for 10 days and mice were euthanized when their aggregate clinical score reached 8 (including a score of 0-3 for weight loss) as detailed in the approved IACUC protocol. Sex as a biological variable not detected. Randomization not detected. Blinding Symptoms were scored in a blinded manner for ruffled fur, hunching and activity, and scored from 0-3 (0 normal, 3 severe) for 10 days and mice were euthanized when their aggregate clinical score reached 8 (including a score of 0-3 for weight loss) as detailed in the approved IACUC protocol. Power Analysis not detected. Cell Line … SciScore for 10.1101/2021.12.06.471483: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: Symptoms were scored in a blinded manner for ruffled fur, hunching and activity, and scored from 0-3 (0 normal, 3 severe) for 10 days and mice were euthanized when their aggregate clinical score reached 8 (including a score of 0-3 for weight loss) as detailed in the approved IACUC protocol. Sex as a biological variable not detected. Randomization not detected. Blinding Symptoms were scored in a blinded manner for ruffled fur, hunching and activity, and scored from 0-3 (0 normal, 3 severe) for 10 days and mice were euthanized when their aggregate clinical score reached 8 (including a score of 0-3 for weight loss) as detailed in the approved IACUC protocol. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Inhibition of RBD/huACE2 interaction: Neutralizing antibodies were assessed using a lateral flow assay that semi-quantitatively measures levels of antibodies that prevent binding of Washington strain RBD to ACE2, as previously described (15). ACE2suggested: NoneExperimental Models: Cell Lines Sentences Resources Viruses: Mouse adapted SARS-CoV-2 SARS2-N501YMA30 was propagated in A549-huACE2 cells (11). A549-huACE2suggested: NoneCell lines: African green monkey kidney Vero cells (E6) or (CCL81) (obtained from ATCC) were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco catalog no. 11965), supplemented with 10% fetal bovine serum (FBS), 100 U/ml of penicillin, 100 μg/ml streptomycin, 50 μg/ml gentamicin, 1mM sodium pyruvate, and 10mM HEPES. Verosuggested: ATCC Cat# CCL-81, RRID:CVCL_0059)Human A549 cells (Verified by ATCC) were cultured in RPMI 1640 (Gibco catalog no. 11875) supplemented with 10% FBS, 100 U/ml of penicillin, and 100 μg/ml streptomycin. A549suggested: NoneThe generation of A549-ACE2 cells was described previously (19). A549-ACE2suggested: NoneExperimental Models: Organisms/Strains Sentences Resources Immunization: BALB/c mice at age 7 weeks were immunized with 106 pfu of NYVAC-KC-pfsSpike. BALB/csuggested: NoneRecombinant DNA Sentences Resources For insertion of foreign genes into the NYVAC-KC genome, we constructed a cassette (pGNR-cmrS) that encodes an E. coli gyrase/PKR fusion protein that confers coumermycin (cmr) sensitivity (14), a neoR gene and expresses GFP (13). pGNR-cmrSsuggested: NoneResults 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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