National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
T32 HL069765
United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
U19 AI142790
United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
F31 AI145189
United States
National Institutes of Health/National Center for Advancing Translational Sciences (NIH/NCATS)
UL1 TR002243
United States
Defense Threat Reduction Agency (DTRA)
HDTRA1-13-1-0034
United States
Citation
Journal: Cell / Year: 2020 Title: Human Antibodies Protect against Aerosolized Eastern Equine Encephalitis Virus Infection. Authors: Lauren E Williamson / Theron Gilliland / Pramod K Yadav / Elad Binshtein / Robin Bombardi / Nurgun Kose / Rachel S Nargi / Rachel E Sutton / Clarissa L Durie / Erica Armstrong / Robert H ...Authors: Lauren E Williamson / Theron Gilliland / Pramod K Yadav / Elad Binshtein / Robin Bombardi / Nurgun Kose / Rachel S Nargi / Rachel E Sutton / Clarissa L Durie / Erica Armstrong / Robert H Carnahan / Lauren M Walker / Arthur S Kim / Julie M Fox / Michael S Diamond / Melanie D Ohi / William B Klimstra / James E Crowe / Abstract: Eastern equine encephalitis virus (EEEV) is one of the most virulent viruses endemic to North America. No licensed vaccines or antiviral therapeutics are available to combat this infection, which has ...Eastern equine encephalitis virus (EEEV) is one of the most virulent viruses endemic to North America. No licensed vaccines or antiviral therapeutics are available to combat this infection, which has recently shown an increase in human cases. Here, we characterize human monoclonal antibodies (mAbs) isolated from a survivor of natural EEEV infection with potent (<20 pM) inhibitory activity of EEEV. Cryo-electron microscopy reconstructions of two highly neutralizing mAbs, EEEV-33 and EEEV-143, were solved in complex with chimeric Sindbis/EEEV virions to 7.2 Å and 8.3 Å, respectively. The mAbs recognize two distinct antigenic sites that are critical for inhibiting viral entry into cells. EEEV-33 and EEEV-143 protect against disease following stringent lethal aerosol challenge of mice with highly pathogenic EEEV. These studies provide insight into the molecular basis for the neutralizing human antibody response against EEEV and can facilitate development of vaccines and candidate antibody therapeutics.
History
Deposition
Jun 26, 2020
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Header (metadata) release
Jan 6, 2021
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Map release
Jan 6, 2021
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Update
Jan 13, 2021
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Current status
Jan 13, 2021
Processing site: RCSB / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
EMPIAR-10890 (Title: Motion corrected micrographs - purified SINV/EEEV particles and recombinant anti-EEEV Fab (EEEV-33) Data size: 111.8 Data #1: Motion corrected micrographs Chimeric Eastern Equine Encephalitis Virus with Fab of EEEV-33 Antibody [micrographs - single frame])
Shell ID: 1 / Name: Eastern equine encephalitis virus / Diameter: 850.0 Å / T number (triangulation number): 4
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Experimental details
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Structure determination
Method
cryo EM
Processing
single particle reconstruction
Aggregation state
3D array
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Sample preparation
Concentration
0.1 mg/mL
Buffer
pH: 7.5 Component:
Concentration
Name
50.0 mM
Tris-HCl
0.1 mM
EDTA
100.0 mM
Sodium Chloride
1.0 x
DPBS
Details: Solutions were made fresh
Grid
Model: Homemade / Material: COPPER / Mesh: 400 / Support film - topology: LACEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR
Vitrification
Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV / Details: blot for 10 seconds before plunging.
Details
Purified SINV/EEEV particles and recombinant Fab mixture
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Electron microscopy
Microscope
FEI TITAN
Details
Preliminary grid screening was performed manually
Image recording
Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Frames/image: 1-50 / Number grids imaged: 1 / Number real images: 2200 / Average exposure time: 10.0 sec. / Average electron dose: 50.0 e/Å2
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
In the structure databanks used in Yorodumi, some data are registered as the other names, "COVID-19 virus" and "2019-nCoV". Here are the details of the virus and the list of structure data.
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