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- EMDB-7003: Eilat virus/Venezuelan equine encephalitis virus chimeric vaccine... -

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Basic information

Entry
Database: EMDB / ID: EMD-7003
TitleEilat virus/Venezuelan equine encephalitis virus chimeric vaccine candidate
Map dataCombined, filtered, unmasked map of Eilat virus/VEEV chimera
SampleEilat virus/Venezuelan equine encephalitis virus chimera != Eilat virus

Eilat virus/Venezuelan equine encephalitis virus chimera

  • Virus: Eilat virus
Biological speciesEilat virus
Methodsingle particle reconstruction / cryo EM / Resolution: 8.4 Å
AuthorsKaelber JT / Erasmus JH / Weaver SC / Nasar F / Chiu W
Funding support United States, 2 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)P41GM103832 United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)AI120942 United States
CitationJournal: J Virol / Year: 2018
Title: Novel Insect-Specific Eilat Virus-Based Chimeric Vaccine Candidates Provide Durable, Mono- and Multivalent, Single-Dose Protection against Lethal Alphavirus Challenge.
Authors: Jesse H Erasmus / Robert L Seymour / Jason T Kaelber / Dal Y Kim / Grace Leal / Michael B Sherman / Ilya Frolov / Wah Chiu / Scott C Weaver / Farooq Nasar /
Abstract: Most alphaviruses are mosquito borne and exhibit a broad host range, infecting many different vertebrates, including birds, rodents, equids, humans, and nonhuman primates. Recently, a host- ...Most alphaviruses are mosquito borne and exhibit a broad host range, infecting many different vertebrates, including birds, rodents, equids, humans, and nonhuman primates. Recently, a host-restricted, mosquito-borne alphavirus, Eilat virus (EILV), was described with an inability to infect vertebrate cells based on defective attachment and/or entry, as well as a lack of genomic RNA replication. We investigated the utilization of EILV recombinant technology as a vaccine platform against eastern (EEEV) and Venezuelan equine encephalitis viruses (VEEV), two important pathogens of humans and domesticated animals. EILV chimeras containing structural proteins of EEEV or VEEV were engineered and successfully rescued in cells. Cryo-electron microscopy reconstructions at 8 and 11 Å of EILV/VEEV and EILV/EEEV, respectively, showed virion and glycoprotein spike structures similar to those of VEEV-TC83 and other alphaviruses. The chimeras were unable to replicate in vertebrate cell lines or in brains of newborn mice when injected intracranially. Histopathologic examinations of the brain tissues showed no evidence of pathological lesions and were indistinguishable from those of mock-infected animals. A single-dose immunization of either monovalent or multivalent EILV chimera(s) generated neutralizing antibody responses and protected animals against lethal challenge 70 days later. Lastly, a single dose of monovalent EILV chimeras generated protective responses as early as day 1 postvaccination and partial or complete protection by day 6. These data demonstrate the safety, immunogenicity, and efficacy of novel insect-specific EILV-based chimeras as potential EEEV and VEEV vaccines. Mostly in the last decade, insect-specific viruses have been discovered in several arbovirus families. However, most of these viruses are not well studied and largely have been ignored. We explored the use of the mosquito-specific alphavirus EILV as an alphavirus vaccine platform in well-established disease models for eastern (EEE) and Venezuelan equine encephalitis (VEE). EILV-based chimeras replicated to high titers in a mosquito cell line yet retained their host range restriction in vertebrates both and In addition, the chimeras generated immune responses that were higher than those of other human and/or equine vaccines. These findings indicate the feasibility of producing a safe, efficacious, mono- or multivalent vaccine against the encephalitic alphaviruses VEEV and EEEV. Lastly, these data demonstrate how host-restricted, insect-specific viruses can be engineered to develop vaccines against related pathogenic arboviruses that cause severe disease in humans and domesticated animals.
History
DepositionAug 31, 2017-
Header (metadata) releaseSep 13, 2017-
Map releaseNov 22, 2017-
UpdateJan 29, 2020-
Current statusJan 29, 2020Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.175
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by radius
  • Surface level: 0.175
  • Imaged by UCSF Chimera
  • Download
Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_7003.png

Downloads & links

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Map

FileDownload / File: emd_7003.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationCombined, filtered, unmasked map of Eilat virus/VEEV chimera
Voxel sizeX=Y=Z: 2.326 Å
Density
Contour LevelBy AUTHOR: 0.175 / Movie #1: 0.175
Minimum - Maximum-0.48278403 - 1.2507095
Average (Standard dev.)0.0031908017 (±0.066673)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin-192-192-192
Dimensions384384384
Spacing384384384
CellA=B=C: 893.18396 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z2.3262.3262.326
M x/y/z384384384
origin x/y/z0.0000.0000.000
length x/y/z893.184893.184893.184
α/β/γ90.00090.00090.000
start NX/NY/NZ-38-19-20
NX/NY/NZ858082
MAP C/R/S123
start NC/NR/NS-192-192-192
NC/NR/NS384384384
D min/max/mean-0.4831.2510.003

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Supplemental data

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Sample components

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Entire : Eilat virus/Venezuelan equine encephalitis virus chimera

EntireName: Eilat virus/Venezuelan equine encephalitis virus chimera
Components
  • Virus: Eilat virus

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Supramolecule #1: Eilat virus

SupramoleculeName: Eilat virus / type: virus / ID: 1 / Parent: 0 / NCBI-ID: 1231903 / Sci species name: Eilat virus / Virus type: VIRION / Virus isolate: OTHER / Virus enveloped: Yes / Virus empty: No
Host (natural)Organism: Aedes (mosquito)
Host systemOrganism: Aedes albopictus (Asian tiger mosquito) / Recombinant cell: C7/10
Molecular weightTheoretical: 42 MDa

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 7.4
Component:
ConcentrationFormulaName
0.05 molarC4H11NO3Tris
0.1 molarNaClSodium chloridesodium chloride
0.001 molarC10H16N2O8Ethylenediaminetetraacetic acid
GridModel: Quantifoil / Material: COPPER / Mesh: 200
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeJEOL 3200FSC
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 4.7 mm / Nominal magnification: 25000
Specialist opticsEnergy filter - Name: In-column Omega Filter
Sample stageSpecimen holder model: JEOL 3200FSC CRYOHOLDER / Cooling holder cryogen: NITROGEN
TemperatureMin: 86.7 K
Image recordingFilm or detector model: DIRECT ELECTRON DE-20 (5k x 3k) / Detector mode: COUNTING / Number real images: 100 / Average exposure time: 1.5 sec. / Average electron dose: 25.0 e/Å2

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Image processing

CTF correctionSoftware - Name: EMAN
Startup modelType of model: OTHER
Details: Model generated from an independent data set collected on JEM-2010F and refined in EMAN2.
Initial angle assignmentType: COMMON LINE / Software - Name: MPSA
Final angle assignmentType: COMMON LINE / Software - Name: MPSA
Final reconstructionApplied symmetry - Point group: I (icosahedral) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 8.4 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: MPSA / Number images used: 4760
FSC plot (resolution estimation)

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