Vaccinia virus-based vaccines confer protective immunity against SARS-CoV-2 virus in Syrian hamsters

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1. Version published to 10.1371/journal.pone.0257191

   Sep 9, 2021
2. 

   ScreenIT

   Aug 5, 2021

   SciScore for 10.1101/2021.08.03.454910: (What is this?)

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

   Table 1: Rigor

   Ethics	Field Sample Permit: All animal protocols were approved by the Institutional Animal Care and Utilization Committee of Academia Sinica and were conducted in strict accordance with the guidelines on animal use and care of the Taiwan National Research Council’s Guide. IACUC: All animal protocols were approved by the Institutional Animal Care and Utilization Committee of Academia Sinica and were conducted in strict accordance with the guidelines on animal use and care of the Taiwan National Research Council’s Guide.
   Sex as a biological variable	Eight-week-old female C57BL/6 mice (Charles River strain) were purchased from BioLASCO Taiwan Co. Ltd.
   Randomization	not detected.
   Blinding	not detected.
   Power …

   SciScore for 10.1101/2021.08.03.454910: (What is this?)

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

   Table 1: Rigor

   Ethics	Field Sample Permit: All animal protocols were approved by the Institutional Animal Care and Utilization Committee of Academia Sinica and were conducted in strict accordance with the guidelines on animal use and care of the Taiwan National Research Council’s Guide. IACUC: All animal protocols were approved by the Institutional Animal Care and Utilization Committee of Academia Sinica and were conducted in strict accordance with the guidelines on animal use and care of the Taiwan National Research Council’s Guide.
   Sex as a biological variable	Eight-week-old female C57BL/6 mice (Charles River strain) were purchased from BioLASCO Taiwan Co. Ltd.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Then, the cells were washed with PBS and stained for the secondary antibody FITC- conjugated goat anti-Rabbit IgG Ab (F1262, 1:500 dilution) for 1 h at room temperature, followed by staining with DAPI (5 μg/ml, D21490, Molecular Probes) for 5 min and mounting with Vectashield mounting solution (H-1000, Vector Laboratories).	anti-Rabbit IgG suggested: (Sigma-Aldrich Cat# F1262, RRID:AB_259430) F1262 suggested: (Sigma-Aldrich Cat# F1262, RRID:AB_259430) Vectashield mounting solution (H-1000 suggested: None
   These blots were then washed three times with PBST, incubated at r.t. with HRP goat anti-mouse (31430, 1:20,000) or HRP goat anti-hamster (PA1-28823, 1:5,000) antibodies for 1 h at r.t. and then developed using a Western Lightning Enhanced Chemiluminescence kit (PerkinElmer) according to the manufacturer’s protocol.	anti-mouse (31430 suggested: (Thermo Fisher Scientific Cat# 31430, RRID:AB_228307) anti-hamster (PA1-28823 suggested: (Thermo Fisher Scientific Cat# PA1-28823, RRID:AB_10986856)
   Cells were incubated with SARS-CoV-2 anti- RBD antibody (40592-T62, 1:500) at 4°C for 1 h.	anti- RBD suggested: None
   To detect anti-spike antibody in the sera of immunized mice and hamsters, SF9 insect cells were infected with either wild type baculovirus (WT-BAC) or a recombinant baculovirus (S-BAC) that expressed a chimeric SARS-CoV-2 S-gp64 protein in which the transmembrane and C-terminal regions of S protein were replaced by the transmembrane and C-terminal regions of baculovirus GP64 so that the S-gp64 fusion protein would be expressed on insect cell surfaces.	anti-spike suggested: None
   SARS-CoV-2 neutralization assay: Serially diluted antibodies from immunized mice or hamsters were incubated at 37 °C for 1 h with 100 TCID50 SARS-CoV-2 TCDC#4 (hCoV-19/Taiwan/4/2020).	hCoV-19/Taiwan/4/2020 suggested: None
   Immunoglobulin ELISA for SARS-CoV-2 S-specific antibodies: Immunoglobulin ELISA was performed as described previously (17) with some modifications.	SARS-CoV-2 suggested: None S-specific suggested: None
   Coated plates were incubated for 1 h at r.t. with sera that had been serially diluted in PBS containing 1% BSA, then washed with PBST, and incubated with HRP-conjugated IgG2C (PA1-29288, 1:15000) or HRP-conjugated IgG1 (PA1-74421, 1:6000) secondary antibodies at r.t. for 1 h.	HRP-conjugated IgG2C suggested: None HRP-conjugated IgG1 suggested: None PA1-74421 suggested: (Thermo Fisher Scientific Cat# PA1-74421, RRID:AB_10988195)
   The cells were subsequently washed twice with FACS buffer, and then incubated with an antibody cocktail including anti-CD3-PE/Cyanine7, anti-CD4-FITC, anti-CD8-Pacific blue, anti-CD44- PE and anti-CD62L-APC for 15 min on ice.	anti-CD3-PE/Cyanine7 suggested: None anti-CD4-FITC suggested: (Sigma-Aldrich Cat# F1773, RRID:AB_476965) anti-CD8-Pacific blue , anti-CD44- PE suggested: None anti-CD62L-APC suggested: None
   Immunohistochemical staining was performed with a monoclonal rabbit anti-SARS-CoV/SARS-CoV-2 nucleocapsid (NP) antibody (1:1000, 40143-R001, Sino Biological), followed by incubation with Dako EnVision+ System HRP.	anti-SARS-CoV/SARS-CoV-2 nucleocapsid (NP suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   HuTK-143 cells were cultured in MEM medium supplemented with 10% FBS and 1% PS.	HuTK-143 suggested: None
   SARS-CoV-2 TCDC#4 (hCoV-19/Taiwan/4/2020) is a local isolate and it was propagated on Vero-E6 cells.	Vero-E6 suggested: None
   Immunofluorescence staining of cell surface S protein: BHK21 and BSC40 cells were infected respectively with MVA-S or v-NY-S at a multiplicity of infection (MOI) of 5 PFU/cell for 1 h, washed with PBS, and then incubated in growth media for a further 12 h.	BSC40 suggested: ATCC Cat# CRL-2761, RRID:CVCL_3656)
   To test reactivity of immunized mouse and hamster sera to SARS-CoV-2 spike protein, the extracellular domain of spike protein from residue 14 to 1209, consisting of S1 and S2 but without the transmembrane domain, was expressed in HEK 293 cells and subsequently purified.	HEK 293 suggested: CLS Cat# 300192/p777_HEK293, RRID:CVCL_0045)
   Flow cytometry analysis of cell-surface SARS-CoV-2 S protein expression: To detect spike protein expression on the surface of cells infected with MVA-S or v-NY-S, BSC40 and BHK21 cells (5×105) were infected with v-NY-S and MVA-S, respectively, at an MOI of 5 PFU/cell and incubated for 12 h, before being detached via treatment with 2 mM EDTA in PBS.	BHK21 suggested: None
   The mixture was then added to HEK-293T cells expressing human ACE2 receptor (104 cells/well of a 96-well plate) and incubated for 24 h at 37 °C.	HEK-293T suggested: None
   The mixtures were then added to pre-seeded Vero E6 cells for a 4-day incubation.	Vero E6 suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Prime-boost immunization regimens in mice: Eight-week-old female C57BL/6 mice were housed in the Animal Facility of Academia Sinica (Taipei, Taiwan) for at least 3 days prior to vaccination experiments.	C57BL/6 suggested: None
   Recombinant DNA
   Sentences	Resources
   Construction of recombinant vaccinia viruses: To generate recombinant vaccinia viruses expressing SARS-CoV-2 S protein (Isolate Wuhan-Hu-1, NC_045512), a human codon-optimized open reading frame (ORF) encoding full-length SARS-CoV-2 S protein was inserted into a pSC11 plasmid and under regulatory control by an early and late p7.5k promoter to obtain pSC11-S plasmid (117).	pSC11 suggested: None
   The pSC11-S plasmid was transfected into HuTK-143 cells infected with the wild type v-NY virus strain.	pSC11-S suggested: None
   Software and Algorithms
   Sentences	Resources
   Eight-week-old female C57BL/6 mice (Charles River strain) were purchased from BioLASCO Taiwan Co. Ltd.	BioLASCO suggested: None
   Cells were acquired using a BD LSR II (BD Biosciences) flow cytometer and data analyses were performed with FlowJo 8.7 software.	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   The oligonucleotides were synthesized by Genomics BioSci	Genomics suggested: (UTHSCSA Genomics Core, RRID:SCR_012239)
   Statistical analyses: Statistical analyses were conducted using Student’s t test in Prism (version 9) software (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: We detected the following sentences addressing limitations in the study:

   On the other hand, several limitations of the hamster model exist. SARS-CoV-2 infection in humans affects multiple organ systems including the heart, kidneys, liver, spleen and large intestine (83, 84) whereas in hamsters live virus mainly replicates in respiratory tracts with little evidence of infectious virus in other organs, despite the detection of viral RNA in non-respiratory tissues (55, 85). Furthermore, a lack of hamster-specific biological reagents makes it difficult to characterize immune responses and the mechanisms of pathogenesis in hamsters. A successful vaccine against SARS-CoV-2 should stimulate both humoral and cellular immune responses to inhibit virus replication and prevent disease in the host (86). In both mouse and hamster model our prime-boost immunization with MVA-S and v-NY-S generated high titers of neutralizing antibodies. Evidence from preclinical studies in nonhuman primates and hamsters indicted that vaccine-induced SARS-CoV-2 neutralizing antibodies correlate with protection against lung infection and clinical disease (19, 55, 87). While it is difficult to study T-cell immune response in hamsters, our immunization regiments induced TH1-biased immune response and effector memory CD8+T cells in mice, consistent with other studies using MVA-based SARS-CoV-2 vaccines (41–43, 88). Interestingly, Tan, et al (2021) reported that early induction of interferon-γ secreting SARS-CoV-2 specific T cells correlates with mild disease and rapid virus clearance...

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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.
   • No funding statement was detected.
   • No protocol registration statement was detected.

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

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3. 

   Version published to 10.1101/2021.08.03.454910v1 on bioRxiv

   Aug 3, 2021