Journal: bioRxiv

Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge

doi: 10.1101/2020.06.18.160655

Figure Lengend Snippet: rVSV-ΔG-spike design and generation strategy: (A) A schematic diagram of the genom organization of WT-VSV genome (top diagram) and rVSV-ΔG-spike (bottom diagram). N: Nucleoprotein, P: Phosphoprotein, M: Matrix, L: Large polymerase, G: Glycoprotein, SPIKE: SARS-CoV-2 spike. (B) pVSV-ΔG-spike map; in red, the inserted S gene. (C) Schematic representation of the generation proces of creating rVSV-ΔG-spike vaccine. Infection of BHK-21 cells with MVA-T7, followed by co-transfecti with pVSV-ΔG-spike, and VSV-system accessory plasmids; Transfection of BHK-21 cells with pCAGGS-VSV-G, followed by infection with the supernatant of the primary transfection to create P1; serial passaging to create rVSV-ΔG-spike.

Article Snippet: The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC.

Techniques: Infection, Transfection, Passaging

Journal: bioRxiv

Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge

doi: 10.1101/2020.06.18.160655

Figure Lengend Snippet: Characterization of rVSV-ΔG-spike: (A) A summary of the genome analysis of several passage of rVSV-ΔG-spike, showing Ct values of VSV-G, VSV-N, and SARS-CoV-2-S indicating the elimination of VSV-G over time, together with increased titer. Also, plaques are formed at late passages. (B) Immunofluorescence images of Vero E6 cells infected with early passage (P5)-rVSV-ΔG-spike, or lat passage (P13)-rVSV-ΔG-spike, stained with a SARS-CoV-2 antibody (green) and DAPI for nuclei stainin (blue). Top and bottom panels show insets at low (10X) and high magnification (25x), respectively. rVSV-ΔG-spike at P5 forms syncytia, whereas P13 show individual infected cells, with no evidence of syncytia. (C) Transmission electron microscopy of rVSV-ΔG-spike (top panel) compared to WT-VSV. Right panel shows immunogold labeling against RBD.

Article Snippet: The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC.

Techniques: Immunofluorescence, Infection, Staining, Transmission Assay, Electron Microscopy, Labeling

Journal: bioRxiv

Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge

doi: 10.1101/2020.06.18.160655

Figure Lengend Snippet: Surface antigenic similarity of rVSV-ΔG-spike and SARS-CoV-2: (A) Immunofluorescent images of Vero E6 cells infected with either WT-VSV (left panel), rVSV-ΔG-spike (middle panel), or SARS-CoV-2, stained with serum from COVID-19 human convalescent serum (right panel). (B) Correlation analysis of neutralization of rVSV-ΔG-spike and SARS-CoV-2 by a panel of sera from COVID-19 convalescent patients. For each sera (n=12), NT 50 values were determined for neutralization of rVSV-ΔG-spike, or SARS-CoV-2. The NT 50 values were plotted to determine the correlation between th neutralization assays. R 2 =0.911.

Article Snippet: The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC.

Techniques: Infection, Staining, Neutralization

Journal: bioRxiv

Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge

doi: 10.1101/2020.06.18.160655

Figure Lengend Snippet: Establishment of golden Syrian hamster SARS-CoV-2 model: (A) Body weight changes of hamsters infected with SARS-CoV-2 at either 5×10 4 (n=7), 5×10 5 (n=7), or 5×10 6 (n=10) pfu/hamster, compared to mock-infected hamsters (n=6). p<0.05 = *, p<0.001 = *** (B-G) General histology (H&E) an SARS-CoV-2 Immunolabeling of hamster lungs. Lungs were isolated and processed for paraffi embedding from naive (B, D, F), and SARS-CoV-2 5×10 6 7dpi (C, E, G) hamster groups. Sections (5μm) were taken for H&E staining (B-E) and SARS-CoV-2 immunolabeling (F-G, DAPI-Blue, SARS-CoV-2-Green). Images B-C: magnification-x2, bar= 100μm; Pictures D-E: magnification-x10, bar= 100μm; Pictures F-G: magnification-x60, bar= 10μm. Black arrows indicate patches of focal inflammation, pleural invagination and alveolar collapse. “*”-indicates hemorrhagic areas. “#”-indicates edema and protein ric exudates. Black arrow heads indicate pulmonary mononuclear cells. White arrows indicate SARS-CoV-positive immunolabeling. Naïve group: n=4, SARS-CoV-2 5×10 6 7dpi group: n=1.

Article Snippet: The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC.

Techniques: Infection, Immunolabeling, Isolation, Staining

Journal: bioRxiv

Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge

doi: 10.1101/2020.06.18.160655

Figure Lengend Snippet: Dose-dependent vaccination of hamsters with rVSV-ΔG-spike. (A) Body weight changes of mock-vaccinated hamsters (n=4), and hamsters vaccinated with rVSV-ΔG-spike ranging from 10 4 to 10 8 pfu/hamster (n=8, n=10, n=10, n=10, n=8, for each vaccinated group, respectively). (B) NT 50 values of neutralization of SARS-CoV-2 by sera from hamsters following i.m. vaccination with rVSV-ΔG-spik ranging from 10 4 to 10 8 pfu/hamster. n=4 for each group. Means and SEM are indicated below the graph.

Article Snippet: The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC.

Techniques: Neutralization

Journal: bioRxiv

Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge

doi: 10.1101/2020.06.18.160655

Figure Lengend Snippet: Detection and neutralization of SARS-CoV-2 by sera from hamsters following s.c. vaccinatio with rVSV-ΔG-spike: (A) Immunofluorescence images of Vero E6 cells infected with SARS-CoV-2, stai with sera from either mock-vaccinated hamsters (left panel) or rVSV-ΔG-spike vaccinated-hamsters (right panel). (B) Plaque reduction neutralization test (PRNT) of hamster sera collected from naïve hamster (PBF, n=5) or hamsters vaccinated with rVSV-ΔG-spike 25 days following vaccination (n=5). (C) Bod weight changes of mock-vaccinated hamsters (Mock, n=16), and hamsters vaccinated with rVSV-ΔG-spike (n=8).

Article Snippet: The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC.

Techniques: Neutralization, Immunofluorescence, Infection, Plaque Reduction Neutralization Test

Journal: bioRxiv

Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge

doi: 10.1101/2020.06.18.160655

Figure Lengend Snippet: Single-dose rVSV-ΔG-spike vaccine efficacy in hamsters following SARS-CoV-2 challenge. (A) Body weight changes of hamsters infected with SARS-CoV-2 (n=12), and hamsters vaccinated wit rVSV-ΔG-spike and infected with 5×10 6 pfu/hamster (n=10) 25 days post-vaccination, compared to moc hamsters (n=8). p<0.001 = ***. (B) Viral loads in lungs (5dpi) of hamsters infected with 5×10 6 pfu/hamster of SARS-CoV-2 (n=5), and hamsters vaccinated with 1×10 6 pfu/hamster of rVSV-ΔG-spike, and the infected with 5×10 6 pfu/hamster of SARS-CoV-2 (n=2). Limit of detection (LOD, 500 pfu/lung).

Article Snippet: The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC.

Techniques: Infection

Journal: bioRxiv

Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge

doi: 10.1101/2020.06.18.160655

Figure Lengend Snippet: Histopathology and viral load of rVSV-ΔG-spike vaccinated and infected lungs: General histology (H&E) and SARS-CoV-2 Immunolabeling of hamster lungs with and without pre vaccination. Lungs were isolated and processed for paraffin embedding from Naïve (A, E, I), vaccinated + infected 10 6 5dpi (B, F, J; C, G, K), and SARS-CoV-2 5×10 6 5dpi (D, H, L) and groups. Sections (5μm) were taken for H&E staining (A-H) and SARS-CoV-2 immunolabeling (I-L, DAPI-Blue, SARS-CoV-2-Green). Pictures A-D: magnification-x1, bar= 100μm; Pictures E-H: magnification-x10, bar= 100μm; Pictures I-L: magnification-x60, bar= 10μm. Black arrows indicate patches of focal inflammation, pleural invaginatio and alveolar collapse. “*”-indicates hemorrhagic areas. “#”-indicates edema and protein rich exudates. Black arrow heads indicate pulmonary mononuclear cells. White arrows indicate CoV-2 positiv immunolabeling. Naïve group: n=4, SARS-CoV-2 5×10 6 pfu/animal 5dpi group: n=1, vaccinated+infecte 10 6 pfu/animal 5dpi group: n=2. (M) Tissue/Air space ratio.

Article Snippet: The pVSV-spike expression plasmid was constructed by PCR amplification of the full length human codon optimized spike gene from pCMV3-SARS-Cov-2 spike expression plasmid (Sino Biological, Cat #VG40588-UT) using the following primers: Forward – atcgatctgtttacgcgtcactATGTTTGTGTTCCTGGTGCTGC; Reverse – atgaagaatctggctagcaggatttgagTCAGGTGTAGTGCAGTTTCACTCC.

Techniques: Histopathology, Infection, Immunolabeling, Isolation, Staining