A Single Intranasal Inoculation with a Paramyxovirus-Vectored Vaccine Protects Guinea Pigs against a Lethal-Dose Ebola Virus Challenge

ABSTRACT To determine whether intranasal inoculation with a paramyxovirus-vectored vaccine can induce protective immunity against Ebola virus (EV), recombinant human parainfluenza virus type 3 (HPIV3) was modified to express either the EV structural glycoprotein (GP) by itself (HPIV3/EboGP) or together with the EV nucleoprotein (NP) (HPIV3/EboGP-NP). Expression of EV GP by these recombinant viruses resulted in its efficient incorporation into virus particles and increased cytopathic effect in Vero cells. HPIV3/EboGP was 100-fold more efficiently neutralized by antibodies to EV than by antibodies to HPIV3. Guinea pigs infected with a single intranasal inoculation of 105.3 PFU of HPIV3/EboGP or HPIV3/EboGP-NP showed no apparent signs of disease yet developed a strong humoral response specific to the EV proteins. When these animals were challenged with an intraperitoneal injection of 103 PFU of EV, there were no outward signs of disease, no viremia or detectable EV antigen in the blood, and no evidence of infection in the spleen, liver, and lungs. In contrast, all of the control animals died or developed severe EV disease following challenge. The highly effective immunity achieved with a single vaccine dose suggests that intranasal immunization with live vectored vaccines based on recombinant respiratory viruses may be an advantageous approach to inducing protective responses against severe systemic infections, such as those caused by hemorrhagic fever agents.

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Postexposure Protection of Guinea Pigs against a Lethal Ebola Virus Challenge Is Conferred by RNA Interference

The Journal of Infectious Diseases ◽

10.1086/504267 ◽

2006 ◽

Vol 193 (12) ◽

pp. 1650-1657 ◽

Cited By ~ 172

Author(s):

Thomas W. Geisbert ◽

Lisa E. Hensley ◽

Elliott Kagan ◽

Erik Zhaoying Yu ◽

Joan B. Geisbert ◽

...

Keyword(s):

Rna Interference ◽

Guinea Pigs ◽

Ebola Virus ◽

Virus Challenge

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Successful Topical Respiratory Tract Immunization of Primates against Ebola Virus

Journal of Virology ◽

10.1128/jvi.00105-07 ◽

2007 ◽

Vol 81 (12) ◽

pp. 6379-6388 ◽

Cited By ~ 113

Author(s):

Alexander Bukreyev ◽

Pierre E. Rollin ◽

Mallory K. Tate ◽

Lijuan Yang ◽

Sherif R. Zaki ◽

...

Keyword(s):

Respiratory Tract ◽

Ebola Virus ◽

Protective Efficacy ◽

Hemorrhagic Fever ◽

Surface Glycoprotein ◽

Respiratory Pathogen ◽

Viral Hemorrhagic Fever ◽

Primate Model ◽

Challenge Virus ◽

Virus Challenge

ABSTRACT Ebola virus causes outbreaks of severe viral hemorrhagic fever with high mortality in humans. The virus is highly contagious and can be transmitted by contact and by the aerosol route. These features make Ebola virus a potential weapon for bioterrorism and biological warfare. Therefore, a vaccine that induces both systemic and local immune responses in the respiratory tract would be highly beneficial. We evaluated a common pediatric respiratory pathogen, human parainfluenza virus type 3 (HPIV3), as a vaccine vector against Ebola virus. HPIV3 recombinants expressing the Ebola virus (Zaire species) surface glycoprotein (GP) alone or in combination with the nucleocapsid protein NP or with the cytokine adjuvant granulocyte-macrophage colony-stimulating factor were administered by the respiratory route to rhesus monkeys—in which HPIV3 infection is mild and asymptomatic—and were evaluated for immunogenicity and protective efficacy against a highly lethal intraperitoneal challenge with Ebola virus. A single immunization with any construct expressing GP was moderately immunogenic against Ebola virus and protected 88% of the animals against severe hemorrhagic fever and death caused by Ebola virus. Two doses were highly immunogenic, and all of the animals survived challenge and were free of signs of disease and of detectable Ebola virus challenge virus. These data illustrate the feasibility of immunization via the respiratory tract against the hemorrhagic fever caused by Ebola virus. To our knowledge, this is the first study in which topical immunization through respiratory tract achieved prevention of a viral hemorrhagic fever infection in a primate model.

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Development of a cAdVax-Based Bivalent Ebola Virus Vaccine That Induces Immune Responses against both the Sudan and Zaire Species of Ebola Virus

Journal of Virology ◽

10.1128/jvi.80.6.2738-2746.2006 ◽

2006 ◽

Vol 80 (6) ◽

pp. 2738-2746 ◽

Cited By ~ 28

Author(s):

Danher Wang ◽

Nicholas U. Raja ◽

Charles M. Trubey ◽

Laure Y. Juompan ◽

Min Luo ◽

...

Keyword(s):

Immune Responses ◽

Ebola Virus ◽

Lethal Dose ◽

Hemorrhagic Fever ◽

Sub Saharan Africa ◽

Potential Efficacy ◽

Efficient Induction ◽

Single Complex ◽

Zaire Ebolavirus ◽

Sub Saharan

ABSTRACT Ebola virus (EBOV) causes a severe hemorrhagic fever for which there are currently no vaccines or effective treatments. While lethal human outbreaks have so far been restricted to sub-Saharan Africa, the potential exploitation of EBOV as a biological weapon cannot be ignored. Two species of EBOV, Sudan ebolavirus (SEBOV) and Zaire ebolavirus (ZEBOV), have been responsible for all of the deadly human outbreaks resulting from this virus. Therefore, it is important to develop a vaccine that can prevent infection by both lethal species. Here, we describe the bivalent cAdVaxE(GPs/z) vaccine, which includes the SEBOV glycoprotein (GP) and ZEBOV GP genes together in a single complex adenovirus-based vaccine (cAdVax) vector. Vaccination of mice with the bivalent cAdVaxE(GPs/z) vaccine led to efficient induction of EBOV-specific antibody and cell-mediated immune responses to both species of EBOV. In addition, the cAdVax technology demonstrated induction of a 100% protective immune response in mice, as all vaccinated C57BL/6 and BALB/c mice survived challenge with a lethal dose of ZEBOV (30,000 times the 50% lethal dose). This study demonstrates the potential efficacy of a bivalent EBOV vaccine based on a cAdVax vaccine vector design.

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IgY antibodies against Ebola virus possess post-exposure protection in a murine pseudovirus challenge model and excellent thermostability

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0008403 ◽

2021 ◽

Vol 15 (3) ◽

pp. e0008403

Author(s):

Yuan Zhang ◽

Yanqiu Wei ◽

Yunlong Li ◽

Xuan Wang ◽

Yang Liu ◽

...

Keyword(s):

Thermal Stability ◽

Ebola Virus ◽

Protective Efficacy ◽

High Titer ◽

Lethal Dose ◽

Hemorrhagic Fever ◽

High Dose ◽

Excellent Thermal Stability ◽

Post Exposure

Ebola virus (EBOV) is one of the most virulent pathogens that causes hemorrhagic fever and displays high mortality rates and low prognosis rates in both humans and nonhuman primates. The post-exposure antibody therapies to prevent EBOV infection are considered effective as of yet. However, owing to the poor thermal stability of mammalian antibodies, their application in the tropics has remained limited. Therefore, a thermostable therapeutic antibody against EBOV was developed modelled on the poultry(chicken) immunoglobulin Y (IgY). The IgY antibodies retaining their neutralising activity at 25°C for one year, displayed excellent thermal stability, opposed to conventional polyclonal antibodies (pAbs) or monoclonal antibodies (mAbs). Laying hens were immunised with a variety of EBOV vaccine candidates and it was confirmed that VSVΔG/EBOVGP encoding the EBOV glycoprotein could induce high titer neutralising antibodies against EBOV. The therapeutic efficacy of immune IgY antibodies in vivo was evaluated in the newborn Balb/c mice who have been challenged with the VSVΔG/EBOVGP model. Mice that have been challenged with a lethal dose of the pseudovirus were treated 2 or 24 h post-infection with different doses of anti-EBOV IgY. The group receiving a high dose of 106 NAU/kg (neutralising antibody units/kilogram) showed complete protection with no symptoms of a disease, while the low-dose group was only partially protected. Conversely, all mice receiving naive IgY died within 10 days. In conclusion, the anti-EBOV IgY exhibits excellent thermostability and protective efficacy. Anti-EBOV IgY shows a lot of promise in entering the realm of efficient Ebola virus treatment regimens.

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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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Ebola Virus Uptake into Polarized Cells from the Apical Surface

Viruses ◽

10.3390/v11121117 ◽

2019 ◽

Vol 11 (12) ◽

pp. 1117 ◽

Cited By ~ 1

Author(s):

Meng Hu ◽

Fei Wang ◽

Wei Li ◽

Xiaowei Zhang ◽

Zhiping Zhang ◽

...

Keyword(s):

Viral Entry ◽

Ebola Virus ◽

Apical Cell ◽

Hemorrhagic Fever ◽

Vero Cells ◽

Cell Polarization ◽

Apical Surface ◽

Intracellular Receptor ◽

Ebov Infection ◽

Niemann Pick

Ebola virus (EBOV) causes severe hemorrhagic fever with high mortality rates. EBOV can infect many types of cells. During severe EBOV infection, polarized epithelial and endothelial cells are damaged, which promotes vascular instability and dysregulation. However, the mechanism causing these symptoms is largely unknown. Here, we studied virus infection in polarized Vero C1008 cells grown on semipermeable Transwell by using EGFP-labeled Ebola virus-like particles (VLPs). Our results showed that Ebola VLPs preferred to enter polarized Vero cells from the apical cell surface. Furthermore, we showed that the EBOV receptors TIM-1 and Axl were distributed apically, which could be responsible for mediating efficient apical viral entry. Macropinocytosis and intracellular receptor Niemann–Pick type C1 (NPC1) had no polarized distribution, although they played roles in virus entry. This study provides a new view of EBOV uptake and cell polarization, which facilitates a further understanding of EBOV infection and pathogenesis.

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Recombinant subunit vaccines protect guinea pigs from lethal Ebola virus challenge

Vaccine ◽

10.1016/j.vaccine.2019.06.035 ◽

2019 ◽

Vol 37 (47) ◽

pp. 6942-6950 ◽

Cited By ~ 4

Author(s):

Axel T. Lehrer ◽

Teri-Ann S. Wong ◽

Michael M. Lieberman ◽

Lisa Johns ◽

Liana Medina ◽

...

Keyword(s):

Guinea Pigs ◽

Ebola Virus ◽

Subunit Vaccines ◽

Virus Challenge

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Ebola Virus Transmission in Guinea Pigs

Journal of Virology ◽

10.1128/jvi.02836-14 ◽

2014 ◽

Vol 89 (2) ◽

pp. 1314-1323 ◽

Cited By ~ 36

Author(s):

Gary Wong ◽

Xiangguo Qiu ◽

Jason S. Richardson ◽

Todd Cutts ◽

Brad Collignon ◽

...

Keyword(s):

Guinea Pigs ◽

Direct Contact ◽

Ebola Virus ◽

Lethal Dose ◽

Virus Transmission ◽

Transmission Efficiency ◽

Transmission Model ◽

Lung Pathology ◽

Virus Spread ◽

Virus Shedding

ABSTRACTEbola virus (EBOV) transmission is currently poorly characterized and is thought to occur primarily by direct contact with infectious material; however transmission from swine to nonhuman primates via the respiratory tract has been documented. To establish an EBOV transmission model for performing studies with statistical significance, groups of six guinea pigs (gps) were challenged intranasally (i.n.) or intraperitoneally (i.p.) with 10,000 times the 50% lethal dose (LD50) of gp-adapted EBOV, and naive gps were then introduced as cage mates for contact exposure at 1 day postinfection (p.i.). The animals were monitored for survival and clinical signs of disease and quantitated for virus shedding postexposure. Changes in the duration of contact of naive gps with infected animals were evaluated for their impact on transmission efficiency. Transmission was more efficient from i.n.- than from i.p.-challenged gps, with 17% versus 83% of naive gps surviving exposure, respectively. Virus shedding was detected beginning at 3 days p.i. from both i.n.- and i.p.-challenged animals. Contact duration positively correlated with transmission efficiency, and the abrogation of direct contact between infected and naive animals through the erection of a steel mesh was effective at stopping virus spread, provided that infectious animal bedding was absent from the cages. Histopathological and immunohistochemical findings show that i.n.-infected gps display enhanced lung pathology and EBOV antigen in the trachea, which supports increased virus transmission from these animals. The results suggest that i.n.-challenged gps are more infectious to naive animals than their systemically infected counterparts and that transmission occurs through direct contact with infectious materials, including those transported through air movement over short distances.IMPORTANCEEbola is generally thought to be spread between humans though infectious bodily fluids. However, a study has shown that Ebola can be spread from pigs to monkeys without direct contact. Further studies have been hampered, because an economical animal model for Ebola transmission is not available. To address this, we established a transmission model in guinea pigs and determined the mechanisms behind virus spread. The survival data, in addition to microscopic examination of lung and trachea sections, show that mucosal infection of guinea pigs is an efficient model for Ebola transmission. Virus spread is increased with longer contact times with an infected animal and is possible without direct contact between an infected and a naive host but can be stopped if infectious materials are absent. These results warrant consideration for the development of future strategies against Ebola transmission and for a better understanding of the parameters involved in virus spread.

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

Journal of Virology ◽

10.1128/jvi.78.10.5458-5465.2004 ◽

2004 ◽

Vol 78 (10) ◽

pp. 5458-5465 ◽

Cited By ~ 221

Author(s):

Michael Garbutt ◽

Ryan Liebscher ◽

Victoria Wahl-Jensen ◽

Steven Jones ◽

Peggy Möller ◽

...

Keyword(s):

Ebola Virus ◽

Hemorrhagic Fever ◽

Transcription Unit ◽

Marburg Virus ◽

Lassa Virus ◽

Type I ◽

Pathogenic Potential ◽

Virus Challenge ◽

Transmembrane Glycoprotein ◽

Vesicular Stomatitis

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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ChAdOx1-vectored Lassa fever vaccine elicits a robust cellular and humoral immune response and protects guinea pigs against lethal Lassa virus challenge

npj Vaccines ◽

10.1038/s41541-021-00291-x ◽

2021 ◽

Vol 6 (1) ◽

Author(s):

Robert J. Fischer ◽

Jyothi N. Purushotham ◽

Neeltje van Doremalen ◽

Sarah Sebastian ◽

Kimberly Meade-White ◽

...

Keyword(s):

Single Dose ◽

Guinea Pigs ◽

Lassa Fever ◽

Therapeutic Interventions ◽

Lassa Virus ◽

Glycoprotein Precursor ◽

Lethal Challenge ◽

Virus Challenge ◽

Humoral Immune ◽

Vectored Vaccine

AbstractLassa virus (LASV) infects hundreds of thousands of individuals each year, highlighting the need for the accelerated development of preventive, diagnostic, and therapeutic interventions. To date, no vaccine has been licensed for LASV. ChAdOx1-Lassa-GPC is a chimpanzee adenovirus-vectored vaccine encoding the Josiah strain LASV glycoprotein precursor (GPC) gene. In the following study, we show that ChAdOx1-Lassa-GPC is immunogenic, inducing robust T-cell and antibody responses in mice. Furthermore, a single dose of ChAdOx1-Lassa-GPC fully protects Hartley guinea pigs against morbidity and mortality following lethal challenge with a guinea pig-adapted LASV (strain Josiah). By contrast, control vaccinated animals reached euthanasia criteria 10–12 days after infection. Limited amounts of LASV RNA were detected in the tissues of vaccinated animals. Viable LASV was detected in only one animal receiving a single dose of the vaccine. A prime-boost regimen of ChAdOx1-Lassa-GPC in guinea pigs significantly increased antigen-specific antibody titers and cleared viable LASV from the tissues. These data support further development of ChAdOx1-Lassa-GPC and testing in non-human primate models of infection.

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