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

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Basic information

Database: EMDB / ID: 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:
SourceEilat virus
Methodsingle particle reconstruction / cryo EM / 8.4 Å resolution
AuthorsKaelber JT / Erasmus JH / Weaver SC / Nasar F / Chiu W
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 ...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.
DateDeposition: Aug 31, 2017 / Header (metadata) release: Sep 13, 2017 / Map release: Nov 22, 2017 / Last update: Jul 18, 2018

Structure visualization

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

Downloads & links


Fileemd_7003.map.gz (map file in CCP4 format, 226493 KB)
Projections & slices

Image control

AxesZ (Sec.)Y (Row.)X (Col.)
384 pix
2.33 Å/pix.
= 893.184 Å
384 pix
2.33 Å/pix.
= 893.184 Å
384 pix
2.33 Å/pix.
= 893.184 Å



Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 2.326 Å
Contour Level:0.175 (by author), 0.175 (movie #1):
Minimum - Maximum-0.48278403 - 1.2507095
Average (Standard dev.)0.0031908017 (0.066673)


Space Group Number1
Map Geometry
Axis orderXYZ
CellA=B=C: 893.18396 Å
α=β=γ: 90.0 deg.

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
start NX/NY/NZ
MAP C/R/S123
start NC/NR/NS-192-192-192
D min/max/mean-0.4831.2510.003

Supplemental data

Sample components

Entire Eilat virus/Venezuelan equine encephalitis virus chimera

EntireName: Eilat virus/Venezuelan equine encephalitis virus chimera
Number of components: 1

Component #1: virus, Eilat virus

VirusName: Eilat virus / Class: VIRION / Empty: No / Enveloped: Yes / Isolate: OTHER
MassTheoretical: 42 MDa
SpeciesSpecies: Eilat virus
Source (engineered)Expression System: Aedes albopictus (Asian tiger mosquito) / Cell of expression system: C7/10
Source (natural)Host Species: Aedes

Experimental details

Sample preparation

SpecimenSpecimen state: particle / Method: cryo EM
Sample solutionpH: 7.4
VitrificationInstrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 %

Electron microscopy imaging

ImagingMicroscope: JEOL 3200FSC
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 25 e/Å2 / Illumination mode: FLOOD BEAM
LensMagnification: 25000. X (nominal) / Cs: 4.7 mm / Imaging mode: BRIGHT FIELD / Energy filter: In-column Omega Filter
Specimen HolderModel: JEOL 3200FSC CRYOHOLDER / Temperature: K ( 86.7 - K)
CameraDetector: DIRECT ELECTRON DE-20 (5k x 3k)

Image acquisition

Image acquisitionNumber of digital images: 100

Image processing

ProcessingMethod: single particle reconstruction / Applied symmetry: I (icosahedral) / Number of projections: 4760
3D reconstructionAlgorithm: FOURIER SPACE / Resolution: 8.4 Å / Resolution method: FSC 0.143 CUT-OFF
FSC plot
(resolution estimation)

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