Properties of Replication-Competent Vesicular Stomatitis Virus Vectors Expressing Glycoproteins of Filoviruses and Arenaviruses

ABSTRACT Replication-competent recombinant vesicular stomatitis viruses (rVSVs) expressing the type I transmembrane glycoproteins and selected soluble glycoproteins of several viral hemorrhagic fever agents (Marburg virus, Ebola virus, and Lassa virus) were generated and characterized. All recombinant viruses exhibited rhabdovirus morphology and replicated cytolytically in tissue culture. Unlike the rVSVs with an additional transcription unit expressing the soluble glycoproteins, the viruses carrying the foreign transmembrane glycoproteins in replacement of the VSV glycoprotein were slightly attenuated in growth. Biosynthesis and processing of the foreign glycoproteins were authentic, and the cell tropism was defined by the transmembrane glycoprotein. None of the rVSVs displayed pathogenic potential in animals. The rVSV expressing the Zaire Ebola virus transmembrane glycoprotein mediated protection in mice against a lethal Zaire Ebola virus challenge. Our data suggest that the recombinant VSV can be used to study the role of the viral glycoproteins in virus replication, immune response, and pathogenesis.

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Lassa-Vesicular Stomatitis Chimeric Virus Safely Destroys Brain Tumors

Journal of Virology ◽

10.1128/jvi.00709-15 ◽

2015 ◽

Vol 89 (13) ◽

pp. 6711-6724 ◽

Cited By ~ 15

Author(s):

Guido Wollmann ◽

Eugene Drokhlyansky ◽

John N. Davis ◽

Connie Cepko ◽

Anthony N. van den Pol

Keyword(s):

Brain Tumors ◽

Vesicular Stomatitis Virus ◽

Brain Cancer ◽

Ebola Virus ◽

Marburg Virus ◽

Chimeric Virus ◽

Lassa Virus ◽

Gene Coding ◽

Vesicular Stomatitis ◽

The Brain

ABSTRACTHigh-grade tumors in the brain are among the deadliest of cancers. Here, we took a promising oncolytic virus, vesicular stomatitis virus (VSV), and tested the hypothesis that the neurotoxicity associated with the virus could be eliminated without blocking its oncolytic potential in the brain by replacing the neurotropic VSV glycoprotein with the glycoprotein from one of five different viruses, including Ebola virus, Marburg virus, lymphocytic choriomeningitis virus (LCMV), rabies virus, and Lassa virus. Based onin vitroinfections of normal and tumor cells, we selected two viruses to testin vivo. Wild-type VSV was lethal when injected directly into the brain. In contrast, a novel chimeric virus (VSV-LASV-GPC) containing genes from both the Lassa virus glycoprotein precursor (GPC) and VSV showed no adverse actions within or outside the brain and targeted and completely destroyed brain cancer, including high-grade glioblastoma and melanoma, even in metastatic cancer models. When mice had two brain tumors, intratumoral VSV-LASV-GPC injection in one tumor (glioma or melanoma) led to complete tumor destruction; importantly, the virus moved contralaterally within the brain to selectively infect the second noninjected tumor. A chimeric virus combining VSV genes with the gene coding for the Ebola virus glycoprotein was safe in the brain and also selectively targeted brain tumors but was substantially less effective in destroying brain tumors and prolonging survival of tumor-bearing mice. A tropism for multiple cancer types combined with an exquisite tumor specificity opens a new door to widespread application of VSV-LASV-GPC as a safe and efficacious oncolytic chimeric virus within the brain.IMPORTANCEMany viruses have been tested for their ability to target and kill cancer cells. Vesicular stomatitis virus (VSV) has shown substantial promise, but a key problem is that if it enters the brain, it can generate adverse neurologic consequences, including death. We tested a series of chimeric viruses containing genes coding for VSV, together with a gene coding for the glycoprotein from other viruses, including Ebola virus, Lassa virus, LCMV, rabies virus, and Marburg virus, which was substituted for the VSV glycoprotein gene. Ebola and Lassa chimeric viruses were safe in the brain and targeted brain tumors. Lassa-VSV was particularly effective, showed no adverse side effects even when injected directly into the brain, and targeted and destroyed two different types of deadly brain cancer, including glioblastoma and melanoma.

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Recombinant Vesicular Stomatitis Virus Vector Mediates Postexposure Protection against Sudan Ebola Hemorrhagic Fever in Nonhuman Primates

Journal of Virology ◽

10.1128/jvi.00456-08 ◽

2008 ◽

Vol 82 (11) ◽

pp. 5664-5668 ◽

Cited By ~ 95

Author(s):

Thomas W. Geisbert ◽

Kathleen M. Daddario-DiCaprio ◽

Kinola J. N. Williams ◽

Joan B. Geisbert ◽

Anders Leung ◽

...

Keyword(s):

Vesicular Stomatitis Virus ◽

Rhesus Monkeys ◽

Nonhuman Primates ◽

Ebola Virus ◽

Rhesus Macaques ◽

Hemorrhagic Fever ◽

Marburg Virus ◽

Ebola Hemorrhagic Fever ◽

Zaire Ebolavirus ◽

Vesicular Stomatitis

ABSTRACT Recombinant vesicular stomatitis virus (VSV) vectors expressing homologous filoviral glycoproteins can completely protect rhesus monkeys against Marburg virus when administered after exposure and can partially protect macaques after challenge with Zaire ebolavirus. Here, we administered a VSV vector expressing the Sudan ebolavirus (SEBOV) glycoprotein to four rhesus macaques shortly after exposure to SEBOV. All four animals survived SEBOV challenge, while a control animal that received a nonspecific vector developed fulminant SEBOV hemorrhagic fever and succumbed. This is the first demonstration of complete postexposure protection against an Ebola virus in nonhuman primates and provides further evidence that postexposure vaccination may have utility in treating exposures to filoviruses.

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A Surrogate Animal Model for Screening of Ebola and Marburg Glycoprotein-Targeting Drugs Using Pseudotyped Vesicular Stomatitis Viruses

Viruses ◽

10.3390/v12090923 ◽

2020 ◽

Vol 12 (9) ◽

pp. 923

Author(s):

Takeshi Saito ◽

Junki Maruyama ◽

Noriyo Nagata ◽

Mao Isono ◽

Kosuke Okuya ◽

...

Keyword(s):

Animal Model ◽

Ebola Virus ◽

Antiviral Drug ◽

Passive Immunization ◽

Systemic Infection ◽

Hemorrhagic Fever ◽

Marburg Virus ◽

Therapeutic Effects ◽

Vesicular Stomatitis

Filoviruses, including Ebola virus (EBOV) and Marburg virus (MARV), cause severe hemorrhagic fever in humans and nonhuman primates with high mortality rates. There is no approved therapy against these deadly viruses. Antiviral drug development has been hampered by the requirement of a biosafety level (BSL)-4 facility to handle infectious EBOV and MARV because of their high pathogenicity to humans. In this study, we aimed to establish a surrogate animal model that can be used for anti-EBOV and -MARV drug screening under BSL-2 conditions by focusing on the replication-competent recombinant vesicular stomatitis virus (rVSV) pseudotyped with the envelope glycoprotein (GP) of EBOV (rVSV/EBOV) and MARV (rVSV/MARV), which has been investigated as vaccine candidates and thus widely used in BSL-2 laboratories. We first inoculated mice, rats, and hamsters intraperitoneally with rVSV/EBOV and found that only hamsters showed disease signs and succumbed within 4 days post-infection. Infection with rVSV/MARV also caused lethal infection in hamsters. Both rVSV/EBOV and rVSV/MARV were detected at high titers in multiple organs including the liver, spleen, kidney, and lungs of infected hamsters, indicating acute and systemic infection resulting in fatal outcomes. Therapeutic effects of passive immunization with an anti-EBOV neutralizing antibody were specifically observed in rVSV/EBOV-infected hamsters. Thus, this animal model is expected to be a useful tool to facilitate in vivo screening of anti-filovirus drugs targeting the GP molecule.

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Single-Injection Vaccine Protects Nonhuman Primates against Infection with Marburg Virus and Three Species of Ebola Virus

Journal of Virology ◽

10.1128/jvi.00561-09 ◽

2009 ◽

Vol 83 (14) ◽

pp. 7296-7304 ◽

Cited By ~ 168

Author(s):

Thomas W. Geisbert ◽

Joan B. Geisbert ◽

Anders Leung ◽

Kathleen M. Daddario-DiCaprio ◽

Lisa E. Hensley ◽

...

Keyword(s):

Nonhuman Primates ◽

Ebola Virus ◽

Single Injection ◽

Hemorrhagic Fever ◽

Marburg Virus ◽

Lethal Challenge ◽

Glycoprotein G ◽

Unvaccinated Control ◽

Zaire Ebolavirus ◽

Vesicular Stomatitis

ABSTRACT The filoviruses Marburg virus and Ebola virus cause severe hemorrhagic fever with high mortality in humans and nonhuman primates. Among the most promising filovirus vaccines under development is a system based on recombinant vesicular stomatitis virus (VSV) that expresses a single filovirus glycoprotein (GP) in place of the VSV glycoprotein (G). Here, we performed a proof-of-concept study in order to determine the potential of having one single-injection vaccine capable of protecting nonhuman primates against Sudan ebolavirus (SEBOV), Zaire ebolavirus (ZEBOV), Cote d'Ivoire ebolavirus (CIEBOV), and Marburgvirus (MARV). In this study, 11 cynomolgus monkeys were vaccinated with a blended vaccine consisting of equal parts of the vaccine vectors VSVΔG/SEBOVGP, VSVΔG/ZEBOVGP, and VSVΔG/MARVGP. Four weeks later, three of these animals were challenged with MARV, three with CIEBOV, three with ZEBOV, and two with SEBOV. Three control animals were vaccinated with VSV vectors encoding a nonfilovirus GP and challenged with SEBOV, ZEBOV, and MARV, respectively, and five unvaccinated control animals were challenged with CIEBOV. Importantly, none of the macaques vaccinated with the blended vaccine succumbed to a filovirus challenge. As expected, an experimental control animal vaccinated with VSVΔG/ZEBOVGP and challenged with SEBOV succumbed, as did the positive controls challenged with SEBOV, ZEBOV, and MARV, respectively. All five control animals challenged with CIEBOV became severely ill, and three of the animals succumbed on days 12, 12, and 14, respectively. The two animals that survived CIEBOV infection were protected from subsequent challenge with either SEBOV or ZEBOV, suggesting that immunity to CIEBOV may be protective against other species of Ebola virus. In conclusion, we developed an immunization scheme based on a single-injection vaccine that protects nonhuman primates against lethal challenge with representative strains of all human pathogenic filovirus species.

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Chikungunya and O’nyong-nyong Viruses in Uganda: Implications for Diagnostics

Open Forum Infectious Diseases ◽

10.1093/ofid/ofz001 ◽

2019 ◽

Vol 6 (3) ◽

Cited By ~ 3

Author(s):

Tamara L Clements ◽

Cynthia A Rossi ◽

Amanda K Irish ◽

Hannah Kibuuka ◽

Leigh Anne Eller ◽

...

Keyword(s):

Ebola Virus ◽

Yellow Fever Virus ◽

Rift Valley Fever Virus ◽

Hemorrhagic Fever ◽

Marburg Virus ◽

Fever Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Lassa Virus ◽

Antibody Prevalence ◽

Valley Fever

Abstract Background A serosurvey of healthy blood donors provided evidence of hemorrhagic fever and arthropod-borne virus infections in Uganda. Methods Antibody prevalence to arthropod-borne and hemorrhagic fever viruses in human sera was determined using enzyme-linked immunosorbent assay (ELISA) and plaque reduction neutralization test (PRNT). Results The greatest antibody prevalence determined by ELISA was to chikungunya virus (CHIKV) followed in descending order by West Nile virus (WNV), Crimean-Congo hemorrhagic fever virus (CCHFV), Ebola virus (EBOV), dengue virus (DEN), yellow fever virus (YFV), Rift Valley fever virus (RVFV), Marburg virus (MARV), and Lassa virus (LASV). Further investigation of CHIKV-positive sera demonstrated that the majority of antibody responses may likely be the result of exposure to the closely related alphavirus o’nyong-nyong virus (ONNV). Conclusions As the use of highly specific and sensitive polymerase chain reaction–based assays becomes the diagnostic standard without the corresponding use of the less sensitive but more broadly reactive immunological-based assays, emerging and re-emerging outbreaks will be initially missed, illustrating the need for an orthogonal system for the detection and identification of viruses causing disease.

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Sorting of Marburg Virus Surface Protein and Virus Release Take Place at Opposite Surfaces of Infected Polarized Epithelial Cells

Journal of Virology ◽

10.1128/jvi.75.3.1274-1283.2001 ◽

2001 ◽

Vol 75 (3) ◽

pp. 1274-1283 ◽

Cited By ~ 36

Author(s):

Christian Sänger ◽

Elke Mühlberger ◽

Elena Ryabchikova ◽

Larissa Kolesnikova ◽

Hans-Dieter Klenk ◽

...

Keyword(s):

Epithelial Cells ◽

Apical Membrane ◽

Basolateral Membrane ◽

Surface Protein ◽

Hemorrhagic Fever ◽

Marburg Virus ◽

Virus Surface ◽

Mdckii Cells ◽

Vesicular Stomatitis ◽

Polarized Epithelial Cells

ABSTRACT Marburg virus, a filovirus, causes severe hemorrhagic fever with hitherto poorly understood molecular pathogenesis. We have investigated here the vectorial transport of the surface protein GP of Marburg virus in polarized epithelial cells. To this end, we established an MDCKII cell line that was able to express GP permanently (MDCK-GP). The functional integrity of GP expressed in these cells was analyzed using vesicular stomatitis virus pseudotypes. Further experiments revealed that GP is transported in MDCK-GP cells mainly to the apical membrane and is released exclusively into the culture medium facing the apical membrane. When MDCKII cells were infected with Marburg virus, the majority of GP was also transported to the apical membrane, suggesting that the protein contains an autonomous apical transport signal. Release of infectious progeny virions, however, took place exclusively at the basolateral membrane of the cells. Thus, vectorial budding of Marburg virus is presumably determined by factors other than the surface protein.

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IFN-I Independent Antiviral Immune Response to Vesicular Stomatitis Virus Challenge in Mouse Brain

Vaccines ◽

10.3390/vaccines8020326 ◽

2020 ◽

Vol 8 (2) ◽

pp. 326

Author(s):

Anurag R. Mishra ◽

Siddappa N. Byrareddy ◽

Debasis Nayak

Keyword(s):

Gene Expression ◽

Immune Response ◽

Vesicular Stomatitis Virus ◽

Antiviral Response ◽

Type I ◽

Immune Gene ◽

Antiviral Immune Response ◽

Virus Challenge ◽

Vesicular Stomatitis ◽

Ifn Γ

Type I interferon (IFN-I) plays a pivotal role during viral infection response in the central nervous system (CNS). The IFN-I can orchestrate and regulate most of the innate immune gene expression and myeloid cell dynamics following a noncytopathic virus infection. However, the role of IFN-I in the CNS against viral encephalitis is not entirely clear. Here we have implemented the combination of global differential gene expression profiling followed by bioinformatics analysis to decipher the CNS immune response in the presence and absence of the IFN-I signaling. We observed that vesicular stomatitis virus (VSV) infection induced 281 gene changes in wild-type (WT) mice primarily associated with IFN-I signaling. This was accompanied by an increase in antiviral response through leukocyte vascular patrolling and leukocyte influx along with the expression of potent antiviral factors. Surprisingly, in the absence of the IFN-I signaling (IFNAR−/− mice), a significantly higher (1357) number of genes showed differential expression compared to the WT mice. Critical candidates such as IFN-γ, CCL5, CXCL10, and IRF1, which are responsible for the recruitment of the patrolling leukocytes, are also upregulated in the absence of IFN-I signaling. The computational network analysis suggests the presence of the IFN-I independent pathway that compensates for the lack of IFN-I signaling in the brain. The analysis shows that TNF-α is connected maximally to the networked candidates, thus emerging as a key regulator of gene expression and recruitment of myeloid cells to mount antiviral action. This pathway could potentiate IFN-γ release; thereby, synergistically activating IRF1-dependent ISG expression and antiviral response.

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Differential Immune Responses to Hemorrhagic Fever-Causing Arenaviruses

Vaccines ◽

10.3390/vaccines7040138 ◽

2019 ◽

Vol 7 (4) ◽

pp. 138 ◽

Cited By ~ 4

Author(s):

Mantlo ◽

Paessler ◽

Huang

Keyword(s):

Clinical Importance ◽

Hemorrhagic Fever ◽

Lassa Fever ◽

Innate Immune ◽

Host Immune System ◽

Lassa Virus ◽

Type I ◽

Highly Pathogenic ◽

The Family ◽

Pro Inflammatory Cytokines

The family Arenaviridae contains several pathogens of major clinical importance. The Old World (OW) arenavirus Lassa virus is endemic in West Africa and is estimated to cause up to 300,000 infections each year. The New World (NW) arenaviruses Junín and Machupo periodically cause hemorrhagic fever outbreaks in South America. While these arenaviruses are highly pathogenic in humans, recent evidence indicates that pathogenic OW and NW arenaviruses interact with the host immune system differently, which may have differential impacts on viral pathogenesis. Severe Lassa fever cases are characterized by profound immunosuppression. In contrast, pathogenic NW arenavirus infections are accompanied by elevated levels of Type I interferon and pro-inflammatory cytokines. This review aims to summarize recent findings about interactions of these pathogenic arenaviruses with the innate immune machinery and the subsequent effects on adaptive immunity, which may inform the development of vaccines and therapeutics against arenavirus infections.

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Next-Generation Sequencing Reveals a Controlled Immune Response to Zaire Ebola Virus Challenge in Cynomolgus Macaques Immunized with Vesicular Stomatitis Virus Expressing Zaire Ebola Virus Glycoprotein (VSVΔG/EBOVgp)

Clinical and Vaccine Immunology ◽

10.1128/cvi.00733-14 ◽

2015 ◽

Vol 22 (3) ◽

pp. 354-356 ◽

Cited By ~ 17

Author(s):

Fredrik Barrenas ◽

Richard R. Green ◽

Matthew J. Thomas ◽

G. Lynn Law ◽

Sean C. Proll ◽

...

Keyword(s):

Vesicular Stomatitis Virus ◽

Host Response ◽

Ebola Virus ◽

Mononuclear Cells ◽

Transcriptional Response ◽

Peripheral Blood Mononuclear ◽

Virus Challenge ◽

Cynomolgus Macaques ◽

Vesicular Stomatitis ◽

Generation Sequencing

ABSTRACTVesicular stomatitis virus expressing Zaire Ebola virus (EBOV) glycoprotein (VSVΔG/EBOVgp) could be used as a vaccine to meet the 2014 Ebola virus outbreak. To characterize the host response to this vaccine, we used mRNA sequencing to analyze peripheral blood mononuclear cells (PBMCs) from cynomolgus macaques after VSVΔG/EBOVgp immunization and subsequent EBOV challenge. We found a controlled transcriptional response that transitioned to immune regulation as the EBOV was cleared. This observation supports the safety of the vaccine.

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Macaque Monoclonal Antibodies Targeting Novel Conserved Epitopes within Filovirus Glycoprotein

Journal of Virology ◽

10.1128/jvi.02172-15 ◽

2015 ◽

Vol 90 (1) ◽

pp. 279-291 ◽

Cited By ~ 48

Author(s):

Zhen-Yong Keck ◽

Sven G. Enterlein ◽

Katie A. Howell ◽

Hong Vu ◽

Sergey Shulenin ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Nonhuman Primates ◽

Ebola Virus ◽

Virus Disease ◽

Protective Efficacy ◽

Hemorrhagic Fever ◽

Marburg Virus ◽

Human Pathogens ◽

Content Type ◽

The Core

ABSTRACTFiloviruses cause highly lethal viral hemorrhagic fever in humans and nonhuman primates. Current immunotherapeutic options for filoviruses are mostly specific to Ebola virus (EBOV), although other members ofFiloviridaesuch as Sudan virus (SUDV), Bundibugyo virus (BDBV), and Marburg virus (MARV) have also caused sizeable human outbreaks. Here we report a set of pan-ebolavirus and pan-filovirus monoclonal antibodies (MAbs) derived from cynomolgus macaques immunized repeatedly with a mixture of engineered glycoproteins (GPs) and virus-like particles (VLPs) for three different filovirus species. The antibodies recognize novel neutralizing and nonneutralizing epitopes on the filovirus glycoprotein, including conserved conformational epitopes within the core regions of the GP1 subunit and a novel linear epitope within the glycan cap. We further report the first filovirus antibody binding to a highly conserved epitope within the fusion loop of ebolavirus and marburgvirus species. One of the antibodies binding to the core GP1 region of all ebolavirus species and with lower affinity to MARV GP cross neutralized both SUDV and EBOV, the most divergent ebolavirus species. In a mouse model of EBOV infection, this antibody provided 100% protection when administered in two doses and partial, but significant, protection when given once at the peak of viremia 3 days postinfection. Furthermore, we describe novel cocktails of antibodies with enhanced protective efficacy compared to individual MAbs. In summary, the present work describes multiple novel, cross-reactive filovirus epitopes and innovative combination concepts that challenge the current therapeutic models.IMPORTANCEFiloviruses are among the most deadly human pathogens. The 2014-2015 outbreak of Ebola virus disease (EVD) led to more than 27,000 cases and 11,000 fatalities. While there are five species ofEbolavirusand several strains of marburgvirus, the current immunotherapeutics primarily target Ebola virus. Since the nature of future outbreaks cannot be predicted, there is an urgent need for therapeutics with broad protective efficacy against multiple filoviruses. Here we describe a set of monoclonal antibodies cross-reactive with multiple filovirus species. These antibodies target novel conserved epitopes within the envelope glycoprotein and exhibit protective efficacy in mice. We further present novel concepts for combination of cross-reactive antibodies against multiple epitopes that show enhanced efficacy compared to monotherapy and provide complete protection in mice. These findings set the stage for further evaluation of these antibodies in nonhuman primates and development of effective pan-filovirus immunotherapeutics for use in future outbreaks.

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