Homotypic Non-exclusion by Vesicular Stomatitis Virus in Chick Cell Culture

We report nine full-genome sequences of vesicular stomatitis virus obtained by Illumina next-generation sequencing of RNA, isolated from either cattle epithelial suspensions or cell culture supernatants. Seven of these viral genomes belonged to the New Jersey serotype/species (clade III), while two isolates belonged to the Indiana serotype/species.

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A methyltransferase-defective VSV-based SARS-CoV-2 vaccine candidate provides complete protection against SARS-CoV-2 infection in hamsters

Journal of Virology ◽

10.1128/jvi.00592-21 ◽

2021 ◽

Author(s):

Mijia Lu ◽

Yuexiu Zhang ◽

Piyush Dravid ◽

Anzhong Li ◽

Cong Zeng ◽

...

Keyword(s):

Cell Culture ◽

Immune Responses ◽

Vesicular Stomatitis Virus ◽

Vaccine Candidate ◽

Viral Mrna ◽

S Protein ◽

Vaccine Candidates ◽

Syrian Golden Hamsters ◽

Vesicular Stomatitis ◽

Immunocompromised Mice

The current pandemic of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to dramatic economic and health burdens. Although the worldwide SARS-CoV-2 vaccination campaign has begun, exploration of other vaccine candidates is needed due to the uncertainties of the current approved vaccines such as durability of protection, cross-protection against variant strains, and costs of long-term production, and storage. In this study, we developed a methyltransferase-defective recombinant vesicular stomatitis virus (mtdVSV)-based SARS-CoV-2 vaccine candidate. We generated mtdVSVs expressing SARS-CoV-2 full-length spike (S), S1, or its receptor binding domain (RBD). All these recombinant viruses grew to high titers in mammalian cells despite high attenuation in cell culture. SARS-CoV-2 S protein and its truncations were highly expressed by the mtdVSV vector. These mtdVSV-based vaccine candidates were completely attenuated in both immunocompetent and immunocompromised mice. Among these constructs, mtdVSV-S induced high levels of SARS-CoV-2 specific neutralizing antibodies (NAbs) and Th1-biased T cell immune responses in mice. Syrian golden hamsters immunized with mtdVSV-S triggered SARS-CoV-2 specific NAbs that were higher than convalescent plasma from convalescent COVID-19 patients. In addition, hamsters immunized with mtdVSV-S were completely protected against SARS-CoV-2 replication in lung and nasal turbinate tissues, cytokine storm, and lung pathology. Collectively, our data demonstrate that mtdVSV expressing SARS-CoV-2 S protein is a safe and highly efficacious vaccine candidate against SARS-CoV-2 infection. Significance Viral mRNA cap methyltransferase (MTase) is essential for mRNA stability, protein translation, and innate immune evasion. Thus, viral mRNA cap MTase activity is a novel target for development of live attenuated or live vectored vaccine candidates. Here, we developed a panel of MTase-defective recombinant recombinant vesicular stomatitis virus (mtdVSV)-based SARS-CoV-2 vaccine candidates expressing full-length S, S1, or several versions of the RBD. These mtdVSV-based vaccine candidates grew to high titers in cell culture and were completely attenuated in both immunocompetent and immunocompromised mice. Among these vaccine candidates, mtdVSV-S induces high levels of SARS-CoV-2 specific neutralizing antibody (Nabs) and Th1-biased immune responses in mice. Syrian golden hamsters immunized with mtdVSV-S triggered SARS-CoV-2 specific NAbs that were higher than convalescent plasma from COVID-19 recovered patients. Furthermore, hamsters immunized with mtdVSV-S were completely protected against SARS-CoV-2 challenge. Thus, mtdVSV is a safe and highly effective vector to deliver SARS-CoV-2 vaccine.

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Changes in Cellular Ribonucleic Acid during Growth of Vesicular Stomatitis Virus in Chick Cell Culture

Journal of General Microbiology ◽

10.1099/00221287-21-3-510 ◽

1959 ◽

Vol 21 (3) ◽

pp. 510-518 ◽

Cited By ~ 4

Author(s):

G. TURCO

Keyword(s):

Cell Culture ◽

Vesicular Stomatitis Virus ◽

Ribonucleic Acid ◽

Vesicular Stomatitis

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Synergistic Attenuation of Vesicular Stomatitis Virus by Combination of Specific G Gene Truncations and N Gene Translocations

Journal of Virology ◽

10.1128/jvi.01911-06 ◽

2006 ◽

Vol 81 (4) ◽

pp. 2056-2064 ◽

Cited By ~ 63

Author(s):

David K. Clarke ◽

Farooq Nasar ◽

Margaret Lee ◽

J. Erik Johnson ◽

Kevin Wright ◽

...

Keyword(s):

Cell Culture ◽

Vesicular Stomatitis Virus ◽

Growth Kinetics ◽

Lethal Dose ◽

Reproductive Capacity ◽

Neurological Injury ◽

N Gene ◽

M Gene ◽

Vesicular Stomatitis

ABSTRACT A variety of rational approaches to attenuate growth and virulence of vesicular stomatitis virus (VSV) have been described previously. These include gene shuffling, truncation of the cytoplasmic tail of the G protein, and generation of noncytopathic M gene mutants. When separately introduced into recombinant VSV (rVSV), these mutations gave rise to viruses distinguished from their “wild-type” progenitor by diminished reproductive capacity in cell culture and/or reduced cytopathology and decreased pathogenicity in vivo. However, histopathology data from an exploratory nonhuman primate neurovirulence study indicated that some of these attenuated viruses could still cause significant levels of neurological injury. In this study, additional attenuated rVSV variants were generated by combination of the above-named three distinct classes of mutation. The resulting combination mutants were characterized by plaque size and growth kinetics in cell culture, and virulence was assessed by determination of the intracranial (IC) 50% lethal dose (LD50) in mice. Compared to virus having only one type of attenuating mutation, all of the mutation combinations examined gave rise to virus with smaller plaque phenotypes, delayed growth kinetics, and 10- to 500-fold-lower peak titers in cell culture. A similar pattern of attenuation was also observed following IC inoculation of mice, where differences in LD50 of many orders of magnitude between viruses containing one and two types of attenuating mutation were sometimes seen. The results show synergistic rather than cumulative increases in attenuation and demonstrate a new approach to the attenuation of VSV and possibly other viruses.

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'Shortened Latency' as a Result of Multiple Infection by Vesicular Stomatitis Virus in Chick Cell Culture

Journal of General Microbiology ◽

10.1099/00221287-19-2-340 ◽

1958 ◽

Vol 19 (2) ◽

pp. 340-349 ◽

Cited By ~ 6

Author(s):

P. D. Cooper

Keyword(s):

Cell Culture ◽

Vesicular Stomatitis Virus ◽

Multiple Infection ◽

Vesicular Stomatitis

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In Vivo Replication and Pathogenesis of Vesicular Stomatitis Virus Recombinant M40 Containing Ebola Virus L-Domain Sequences

Infectious Diseases Research and Treatment ◽

10.4137/idrt.s10652 ◽

2012 ◽

Vol 5 ◽

pp. IDRT.S10652

Author(s):

Takashi Irie ◽

Elena Carnero ◽

Adolfo García-Sastre ◽

Ronald N. Harty

Keyword(s):

Cell Culture ◽

Animal Model ◽

Vesicular Stomatitis Virus ◽

Ebola Virus ◽

Small Animal ◽

Small Animal Model ◽

Virus Recombinant ◽

Vesicular Stomatitis ◽

Ptap Motif

The M40 VSV recombinant was engineered to contain overlapping PTAP and PPxY L-domain motifs and flanking residues from the VP40 protein of Ebola virus. Replication of M40 in cell culture is virtually indistinguishable from that of control viruses. However, the presence of the Ebola PTAP motif in the M40 recombinant enabled this virus to interact with and recruit host Tsg101, which was packaged into M40 virions. In this brief report, we compared replication and the pathogenic profiles of M40 and the parental virus M51R in mice to determine whether the presence of the Ebola L-domains and flanking residues altered in vivo characteristics of the virus. Overall, the in vivo characteristics of M40 were similar to those of the parental M51R virus, indicating that the Ebola sequences did not alter pathogenesis of VSV in this small animal model of infection.

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Modifications of the PSAP region of the matrix protein lead to attenuation of vesicular stomatitis virus in vitro and in vivo

Journal of General Virology ◽

10.1099/vir.0.83096-0 ◽

2007 ◽

Vol 88 (9) ◽

pp. 2559-2567 ◽

Cited By ~ 17

Author(s):

Takashi Irie ◽

Elena Carnero ◽

Atsushi Okumura ◽

Adolfo García-Sastre ◽

Ronald N. Harty

Keyword(s):

Cell Culture ◽

Vesicular Stomatitis Virus ◽

Matrix Protein ◽

Virus Assembly ◽

Host Protein ◽

M Protein ◽

Recombinant Viruses ◽

Enhanced Production ◽

The Matrix ◽

Vesicular Stomatitis

The matrix (M) protein of vesicular stomatitis virus (VSV) is a multi-functional protein involved in virus assembly, budding and pathogenesis. The 24PPPY27 late (L) domain of the M protein plays a key role in virus budding, whereas amino acids downstream of the PPPY motif contribute to host protein shut-off and pathogenesis. Using a panel of 37PSAP40 recombinant viruses, it has been demonstrated previously that the PSAP region of M does not possess L-domain activity similar to that of PPPY in BHK-21 cells. This study reports the unanticipated finding that these PSAP recombinants were attenuated in cell culture and in mice compared with control viruses. Indeed, PSAP recombinant viruses exhibited a small-plaque phenotype, reduced CPE, reduced levels of activated caspase-3, enhanced production of IFN-β and reduced titres in the lungs and brains of infected mice. In particular, recombinant virus M6PY>A4-R34E was the most severely attenuated, exhibiting little or no CPE in cell culture and undetectable titres in the lungs and brains of infected mice. These findings indicate an important role for the PSAP region (aa 33–44) of the M protein in the pathology of VSV infection and may have implications for the development of VSV as a vaccine and/or oncolytic vector.

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Vesicular Stomatitis Virus Glycoprotein Is a Determinant of Pathogenesis in Swine, a Natural Host

Journal of Virology ◽

10.1128/jvi.77.14.8039-8047.2003 ◽

2003 ◽

Vol 77 (14) ◽

pp. 8039-8047 ◽

Cited By ~ 34

Author(s):

Isidoro Martinez ◽

Luis L. Rodriguez ◽

Carlos Jimenez ◽

Steven J. Pauszek ◽

Gail W. Wertz

Keyword(s):

Cell Culture ◽

New Jersey ◽

Vesicular Stomatitis Virus ◽

Single Copy ◽

Natural Host ◽

Parental Virus ◽

Vesicular Stomatitis Virus Glycoprotein ◽

Recombinant Viruses ◽

Virus Glycoprotein ◽

Vesicular Stomatitis

ABSTRACT There are two major serotypes of vesicular stomatitis virus (VSV), Indiana (VSIV) and New Jersey (VSNJV). We recovered recombinant VSIVs from engineered cDNAs that contained either (i) one copy of the VSIV G gene (VSIV-GI); (ii) two copies of the G gene, one from each serotype (VSIV-GNJGI); or (iii) a single copy of the GNJ gene instead of the GI gene (VSIV-GNJ). The recombinant viruses expressed the appropriate glycoproteins, incorporated them into virions, and were neutralized by antibodies specific for VSIV (VSIV-GI), VSNJV (VSIV-GNJ), or both (VSIV-GNJGI), according to the glycoprotein(s) they expressed. All recombinant viruses grew to similar titers in cell culture. In mice, VSIV-GNJ and VSIV-GNJGI were attenuated. However, in swine, a natural host for VSV, the GNJ glycoprotein-containing viruses caused more severe lesions and replicated to higher titers than the parental virus, VSIV-GI. These observations implicate the glycoprotein as a determinant of VSV virulence in a natural host and emphasize the differences in VSV pathogenesis between mice and swine.

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A Viable Recombinant Rhabdovirus Lacking Its Glycoprotein Gene and Expressing Influenza Virus Hemagglutinin and Neuraminidase Is a Potent Influenza Vaccine

Journal of Virology ◽

10.1128/jvi.03246-14 ◽

2014 ◽

Vol 89 (5) ◽

pp. 2820-2830 ◽

Cited By ~ 14

Author(s):

Alex B. Ryder ◽

Linda Buonocore ◽

Leatrice Vogel ◽

Raffael Nachbagauer ◽

Florian Krammer ◽

...

Keyword(s):

Cell Culture ◽

Influenza Virus ◽

Influenza Vaccine ◽

Vesicular Stomatitis Virus ◽

Recombinant Virus ◽

Vaccine Candidate ◽

Influenza Vaccines ◽

Glycoprotein Gene ◽

Influenza Virus Hemagglutinin ◽

Vesicular Stomatitis

ABSTRACTThe emergence of novel influenza viruses that cause devastating human disease is an ongoing threat and serves as an impetus for the continued development of novel approaches to influenza vaccines. Influenza vaccine development has traditionally focused on producing humoral and/or cell-mediated immunity, often against the viral surface glycoproteins hemagglutinin (HA) and neuraminidase (NA). Here, we describe a new vaccine candidate that utilizes a replication-defective vesicular stomatitis virus (VSV) vector backbone that lacks the native G surface glycoprotein gene (VSVΔG). The expression of the H5 HA of an H5N1 highly pathogenic avian influenza virus (HPAIV), A/Vietnam/1203/04 (VN1203), and the NA of the mouse-adapted H1N1 influenza virus A/Puerto Rico/8/34 (PR8) in the VSVΔG vector restored the ability of the recombinant virus to replicate in cell culture, without the requirement for the addition of trypsin. We show here that this recombinant virus vaccine candidate was nonpathogenic in mice when given by either the intramuscular or intranasal route of immunization and that thein vivoreplication of VSVΔG-H5N1 is profoundly attenuated. This recombinant virus also provided protection against lethal H5N1 infection after a single dose. This novel approach to vaccination against HPAIVs may be widely applicable to other emerging strains of influenza virus.IMPORTANCEPreparation for a potentially catastrophic influenza pandemic requires novel influenza vaccines that are safe, can be produced and administered quickly, and are effective, both soon after administration and for a long duration. We have created a new influenza vaccine that utilizes an attenuated vesicular stomatitis virus (VSV) vector, to deliver and express influenza virus proteins against which vaccinated animals develop potent antibody responses. The influenza virus hemagglutinin and neuraminidase proteins, expressed on the surface of VSV particles, allowed this vaccine to grow in cell culture and induced a potent antibody response in mice that was effective against infection with a lethal influenza virus. The mice showed no adverse reactions to the vaccine, and they were protected against an otherwise lethal influenza infection after only 14 days postvaccination and after as many as 140 days postvaccination. The ability to rapidly produce this safe and effective vaccine in cell culture is additionally advantageous.

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