Journal: Biosafety and Health

Article Title: Molecular phylogenomic analysis reveals a single origin of monkeypox virus transmission in Guangzhou, China in June 2023

doi: 10.1016/j.bsheal.2023.08.002

Figure Lengend Snippet: Phylogenetic analysis of monkeypox (mpox) virus sequences first reported in Guangzhou, China. A) The eight local sequences (red dot) in FASTA format were aligned by the BioEdit software 7.0.9 with the reference sequences of mpox viral clade IIb downloaded from GISAID (black dot) and NCBI (NC_063383.1, green dot, selected as the outgroup). The maximum likelihood (ML) phylogenetic tree of the full-length genome sequences of mpox virus was constructed with the MEGA software 11.0.11 using the General Time Reversible model with 500 bootstrap replicates. The scale bar represents the genetic distance. Bootstrap values >75% are presented at the corresponding nodes of the tree. Abbreviations: FASTA, Fast-All; GISAID, Global Initiative on Sharing Avian Influenza Data; NCBI, National Center For Biotechnology Information. B) The analysis of amino acid mutation of the eight local sequences compared to the reference Japan|EPI ISL 17692269|2023-04-28. In most cases, the mutation sites are mixed. Only the dominant amino acid is shown. “*” represents termination codon. As for this special site, the CAG (Q) codon accounts for 29.26% of all reads, but the TAG (stop) codon accounts for 70.74%. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: We then performed the amplicon-based mpox viral full-length genome sequencing of mpox virus using a newly developed method by MGI Tech Co., Ltd. (MGI), China.

Techniques: Virus, Software, Construct, Mutagenesis

Journal: bioRxiv

Article Title: Characterization of Morreton Virus (MORV) as a Novel Oncolytic Virotherapy Platform for Liver Cancers

doi: 10.1101/2022.03.10.483848

Figure Lengend Snippet: A, Genome Organization and Ultrastructure of MORV . MORV, a negative-sense RNA Vesiculovirus comprising five major genes (MORV-N, MORV-P, MORV-M, MORV-G, and MORV-L) and 11,181 nucleotides in genome. B, Transmission electron microscopy image of MORV virus, a bullet-shaped virus of 200 nm (length) and 75 nm (width). Defective interfering (DI) particles designated as incomplete particles (IP) were found after several plaque purifications. VP (viral particle) and G (glycoprotein).

Article Snippet: The full-length MORV genome (11,181 nucleotides) comprising genes encoding for the nucleoprotein (MORV-N), phosphoprotein (MORV-P), matrix protein (MORV-M), glycoprotein (MORV-G), and RNA-directed RNA polymerase L protein (MORV-L), was synthesized (Genscript, USA) from the laboratory-adapted viral clone of MORV and subcloned into plasmid (pMORV-XN2). pMORV-XN2, along with helper plasmids (pMORV-P, pMORV-N, and pMORV-L) as previously described .

Techniques: Transmission Assay, Electron Microscopy, Virus

Journal: bioRxiv

Article Title: Characterization of Morreton Virus (MORV) as a Novel Oncolytic Virotherapy Platform for Liver Cancers

doi: 10.1101/2022.03.10.483848

Figure Lengend Snippet: Characterization of MORV . A , Left-Viral titers of MORV and vesiculovirus stomatitis virus (VSV). Recombinant MORV and laboratory-adapted VSV were used to infect BHK-21 cells. B , Supernatants from infected cells were collected at different time points, and viral titer was determined using a TCID 50 (50% tissue culture infective dose) method on Vero cells (1.5 x10 4 ). Right-Serial dilutions of MORV and VSV were used to infect a monolayer of Vero cells (5 x10 5 ), then stained 48 hours later with crystal violet to reveal viral plaques. C, A549 (2 x10 4 ), interferon responsive lung cancer cells were pretreated with various concentrations of universal type I interferon-alpha (IFN-α), infected with viruses at a multiplicity of infection (MOI) of 0.01. Cell viability was measured using a colorimetric assay (MTS, Promega USA) after 48-hours post-infection. D and E, Normal human serum, anti-VSV glycoprotein (anti-VSV-G) antibody, serum from patients treated with VSV (PT13-D2, PT14-22) expressing human IFN-beta (VSV-hIFN-β) were evaluated for their ability to neutralize 400 TCID 50 units of MORV and VSV in Vero cells (2 x10 4 ). Data are expressed as means of triplicate from three independent experiments.

Article Snippet: The full-length MORV genome (11,181 nucleotides) comprising genes encoding for the nucleoprotein (MORV-N), phosphoprotein (MORV-P), matrix protein (MORV-M), glycoprotein (MORV-G), and RNA-directed RNA polymerase L protein (MORV-L), was synthesized (Genscript, USA) from the laboratory-adapted viral clone of MORV and subcloned into plasmid (pMORV-XN2). pMORV-XN2, along with helper plasmids (pMORV-P, pMORV-N, and pMORV-L) as previously described .

Techniques: Virus, Recombinant, Infection, Staining, Colorimetric Assay, Expressing

Journal: bioRxiv

Article Title: Characterization of Morreton Virus (MORV) as a Novel Oncolytic Virotherapy Platform for Liver Cancers

doi: 10.1101/2022.03.10.483848

Figure Lengend Snippet: Cytopathic Effect of MORV and VSV In a Panel of Liver Cancer Cells . A , Monolayers (1.5×10 5 ) of a transformed cholangiocyte cell line (H69), human CCA ( EGI-1, PAX-42, HuCCT-1, SNU-1079, RBE, GBD-1, CAK-1, SNU-245, SNU-308, SNU869), murine CCA (SB), human hepatocellular carcinoma (Hep3B, HepG2,Huh7, Sk-Hep1) and murine HCC ( R1LWT, R2LWT, HCA-1 and Hepa 1-6) cells were mock-infected or infected with MORV ( A ) and VSV ( B ) at an MOI of 0.01, 0.1 and 1. The percentage of cell viability was determined 72 hours post-infection by MTS assays (Promega, USA). The average of three independent experiments are plotted.

Article Snippet: The full-length MORV genome (11,181 nucleotides) comprising genes encoding for the nucleoprotein (MORV-N), phosphoprotein (MORV-P), matrix protein (MORV-M), glycoprotein (MORV-G), and RNA-directed RNA polymerase L protein (MORV-L), was synthesized (Genscript, USA) from the laboratory-adapted viral clone of MORV and subcloned into plasmid (pMORV-XN2). pMORV-XN2, along with helper plasmids (pMORV-P, pMORV-N, and pMORV-L) as previously described .

Techniques: Transformation Assay, Infection

Journal: bioRxiv

Article Title: Characterization of Morreton Virus (MORV) as a Novel Oncolytic Virotherapy Platform for Liver Cancers

doi: 10.1101/2022.03.10.483848

Figure Lengend Snippet: Assessment of MORV and VSV Infectivity and Cytotoxicity In Low Density Lipoprotein Receptor (LDLR) Knockout Cell Line (HAP-1). A, Expression of low-density lipoprotein receptor (LDLR) was measured by flow cytometry using fluorescein (FITC) anti-human LDLR antibody (solid line) or an isotype control antibody (dashed line). B, we infected HAP1 WT or LDLR KO cells (1×10 4 ) cells with different MOIs (10,1, 0.1, 0.01, 0.001, and 0.0001) of MORV or VSV, and 72 hours post-infection, cell viability was measured by MTS assay. C, Representative images of infected or mock-infected HAP1 WT and LDLR KO cells fixed at 48 hours, stained with crystal violet (scale bar=200 µm). Data are shown as mean ± SEM from three independent experiments.

Article Snippet: The full-length MORV genome (11,181 nucleotides) comprising genes encoding for the nucleoprotein (MORV-N), phosphoprotein (MORV-P), matrix protein (MORV-M), glycoprotein (MORV-G), and RNA-directed RNA polymerase L protein (MORV-L), was synthesized (Genscript, USA) from the laboratory-adapted viral clone of MORV and subcloned into plasmid (pMORV-XN2). pMORV-XN2, along with helper plasmids (pMORV-P, pMORV-N, and pMORV-L) as previously described .

Techniques: Infection, Knock-Out, Expressing, Flow Cytometry, Control, MTS Assay, Staining

Journal: bioRxiv

Article Title: Characterization of Morreton Virus (MORV) as a Novel Oncolytic Virotherapy Platform for Liver Cancers

doi: 10.1101/2022.03.10.483848

Figure Lengend Snippet: Intranasal Administration of MORV and VSV in Immunocompetent Mice . A and B, Measurement of body weight and C , Pictures of Hematoxylin and eosin (H&E) stained brain sections from mice treated with high dose of MORV and VSV (1×10 10 TCID 50 /kg). C, complete blood count (white blood cells (WBC) and lymphocytes) values following intranasal administration of increasing doses (1×10 7 , 1×10 8 , 1×10 9 and 1×10 10 TCID 50 /kg) of MORV and VSV.

Article Snippet: The full-length MORV genome (11,181 nucleotides) comprising genes encoding for the nucleoprotein (MORV-N), phosphoprotein (MORV-P), matrix protein (MORV-M), glycoprotein (MORV-G), and RNA-directed RNA polymerase L protein (MORV-L), was synthesized (Genscript, USA) from the laboratory-adapted viral clone of MORV and subcloned into plasmid (pMORV-XN2). pMORV-XN2, along with helper plasmids (pMORV-P, pMORV-N, and pMORV-L) as previously described .

Techniques: Staining

Journal: bioRxiv

Article Title: Characterization of Morreton Virus (MORV) as a Novel Oncolytic Virotherapy Platform for Liver Cancers

doi: 10.1101/2022.03.10.483848

Figure Lengend Snippet: In Vivo Anti-tumor Efficacy of MORV and VSV in Xenograft Mouse Models of Cholangiocarcinoma (CCA) and Hepatocellular Carcinoma (HCC). Single or multiple doses (2) of 1 × 10 7 TCID 50 of MORV or VSV were injected intratumorally into HuCCT-1(CCA) and Hep3B (HCC) tumor-bearing mice. Analysis of the effect of MORV and VSV on tumor growth and tumor inhibition in HuCCT-1 ( A ), and Hep3B ( B ) xenografts.

Article Snippet: The full-length MORV genome (11,181 nucleotides) comprising genes encoding for the nucleoprotein (MORV-N), phosphoprotein (MORV-P), matrix protein (MORV-M), glycoprotein (MORV-G), and RNA-directed RNA polymerase L protein (MORV-L), was synthesized (Genscript, USA) from the laboratory-adapted viral clone of MORV and subcloned into plasmid (pMORV-XN2). pMORV-XN2, along with helper plasmids (pMORV-P, pMORV-N, and pMORV-L) as previously described .

Techniques: In Vivo, Injection, Inhibition

Journal: bioRxiv

Article Title: Characterization of Morreton Virus (MORV) as a Novel Oncolytic Virotherapy Platform for Liver Cancers

doi: 10.1101/2022.03.10.483848

Figure Lengend Snippet: MORV Reduced Tumor Burden At a Lower Dose Compared to VSV In Murine CCA. A, Overview of the in vivo experiment . B, Tumor weight of mice treated with phosphate-buffered saline (PBS), MORV, and VSV at 1 x10 7 TCID 50 or 1×10 8 TCID 50 . C, Frequency of tumor-infiltrating CD3+ CD8+ CTLs. D, E, F and G, graphs showing changes in serum ALP, ALT, IgM and IgG2b.

Article Snippet: The full-length MORV genome (11,181 nucleotides) comprising genes encoding for the nucleoprotein (MORV-N), phosphoprotein (MORV-P), matrix protein (MORV-M), glycoprotein (MORV-G), and RNA-directed RNA polymerase L protein (MORV-L), was synthesized (Genscript, USA) from the laboratory-adapted viral clone of MORV and subcloned into plasmid (pMORV-XN2). pMORV-XN2, along with helper plasmids (pMORV-P, pMORV-N, and pMORV-L) as previously described .

Techniques: In Vivo, Saline