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

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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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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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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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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Ebola and Marburg Viruses

Journal of Disaster Research ◽

10.20965/jdr.2011.p0381 ◽

2011 ◽

Vol 6 (4) ◽

pp. 381-389 ◽

Cited By ~ 7

Author(s):

Eri Nakayama ◽

◽

Ayato Takada

Keyword(s):

Nonhuman Primates ◽

Ebola Virus ◽

Hemorrhagic Fever ◽

Disease Outbreak ◽

Marburg Virus ◽

Virus Group ◽

Imported Cases ◽

Control Disease ◽

Animal Reservoirs ◽

Biosafety Level

Ebola and Marburg viruses, members of the filovirus family, cause severe hemorrhagic fever in human and nonhuman primates and are classified as biosafety level 4 agents. No effective filovirus-specific prophylaxis or treatment is yet commercially available. Filovirus species vary genetically, with one in the Marburg virus group and five in the Ebola virus group. Epidemiological efforts to prevent outbreaks lie mainly in identifying natural animal reservoirs. Increasingly frequent outbreaks in Africa and concerns about bioterrorism and imported cases in nonendemic areas point to the importance of public health in two ways – finding strategies to control disease outbreak and developing effective vaccines and drugs.

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Supramolecular fluorogenic peptide sensor array based on graphene oxide for the differential sensing of ebola virus

Chemical Communications ◽

10.1039/c9cc09981f ◽

2020 ◽

Vol 56 (43) ◽

pp. 5735-5738 ◽

Cited By ~ 2

Author(s):

Meng-Qi Fu ◽

Xu-Chen Wang ◽

Wei-Tao Dou ◽

Guo-Rong Chen ◽

Tony D. James ◽

...

Keyword(s):

Principal Component Analysis ◽

Graphene Oxide ◽

Vesicular Stomatitis Virus ◽

Sensor Array ◽

Ebola Virus ◽

Principal Component ◽

Component Analysis ◽

Marburg Virus ◽

Vesicular Stomatitis ◽

Fluorogenic Peptide

Principal component analysis of a fluorescent supramolecular sensor array based on graphene oxide can be used to differentiate ebola virus from marburg virus and receptor-extensive vesicular stomatitis virus.

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Identification and characterization of defective viral genomes in Ebola virus infected rhesus macaques

Journal of Virology ◽

10.1128/jvi.00714-21 ◽

2021 ◽

Author(s):

Rebecca I. Johnson ◽

Beata Boczkowska ◽

Kendra Alfson ◽

Taylor Weary ◽

Heather Menzie ◽

...

Keyword(s):

Nonhuman Primates ◽

Ebola Virus ◽

Sequence Data ◽

Rna Virus ◽

Hemorrhagic Fever ◽

Marburg Virus ◽

Immunostimulatory Activity ◽

Viral Genomes

Ebola virus (EBOV), of the family Filoviridae, is an RNA virus that can cause hemorrhagic fever with a high mortality rate. Defective viral genomes (DVGs) are truncated genomes that have been observed during multiple RNA virus infections, including  in vitro EBOV infection, and have previously been associated with viral persistence and immunostimulatory activity. As DVGs have been detected in cells persistently infected with EBOV, we hypothesized that DVGs may also accumulate during viral replication in filovirus-infected hosts. Therefore, we interrogated sequence data from serum and tissues using a bioinformatics tool in order to identify the presence of DVGs in nonhuman primates (NHPs) infected with EBOV, Sudan virus (SUDV) or Marburg virus (MARV). Multiple 5’ copy-back DVGs (cbDVGs) were detected in NHP serum during the acute phase of filovirus infection. While the relative abundance of total DVGs in most animals was low, serum collected during acute EBOV and SUDV infections, but not MARV infection, contained a higher proportion of short trailer sequence cbDVGs than the challenge stock. This indicated an accumulation of these DVGs throughout infection, potentially due to the preferential replication of short DVGs over the longer viral genome. Using RT-PCR and deep sequencing, we also confirmed the presence of 5’ cbDVGs in EBOV-infected NHP testes, which is of interest due to EBOV persistence in semen of male survivors of infection. This work suggests that DVGs play a role in EBOV infection in vivo and further study will lead to a better understanding of EBOV pathogenesis. Importance The study of filovirus pathogenesis is critical for understanding the consequences of infection and the development of strategies to ameliorate future outbreaks. Defective viral genomes (DVGs) have been detected during EBOV infections in vitro , however their presence in in vivo infections remains unknown. In this study, DVGs were detected in samples collected from EBOV- and SUDV-infected nonhuman primates (NHPs). The accumulation of these DVGs in the trailer region of the genome during infection indicates a potential role in EBOV and SUDV pathogenesis. In particular, the presence of DVGs in the testes of infected NHPs requires further investigation as it may be linked to the establishment of persistence.

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Vaccine To Confer to Nonhuman Primates Complete Protection against Multistrain Ebola and Marburg Virus Infections

Clinical and Vaccine Immunology ◽

10.1128/cvi.00431-07 ◽

2008 ◽

Vol 15 (3) ◽

pp. 460-467 ◽

Cited By ~ 91

Author(s):

Dana L. Swenson ◽

Danher Wang ◽

Min Luo ◽

Kelly L. Warfield ◽

Jan Woraratanadharm ◽

...

Keyword(s):

Nonhuman Primates ◽

Ebola Virus ◽

De Novo ◽

Lethal Dose ◽

Hemorrhagic Fever ◽

Mortality Rates ◽

Marburg Virus ◽

Virus Infections ◽

Biological Attack ◽

Protection Against Infection

ABSTRACT Filoviruses (Ebola and Marburg viruses) are among the deadliest viruses known to mankind, with mortality rates nearing 90%. These pathogens are highly infectious through contact with infected body fluids and can be easily aerosolized. Additionally, there are currently no licensed vaccines available to prevent filovirus outbreaks. Their high mortality rates and infectious capabilities when aerosolized and the lack of licensed vaccines available to prevent such infectious make Ebola and Marburg viruses serious bioterrorism threats, placing them both on the category A list of bioterrorism agents. Here we describe a panfilovirus vaccine based on a complex adenovirus (CAdVax) technology that expresses multiple antigens from five different filoviruses de novo. Vaccination of nonhuman primates demonstrated 100% protection against infection by two species of Ebola virus and three Marburg virus subtypes, each administered at 1,000 times the lethal dose. This study indicates the feasibility of vaccination against all current filovirus threats in the event of natural hemorrhagic fever outbreak or biological attack.

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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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