|Entry||Database: EMDB / ID: EMD-20888|
|Title||Negative-stain EM map from 3D sorting of BG505-immunized rhesus macaque 4O9 wk 10 serum fab complexed with BG505 SOSIP.664 trimer|
|Sample||Polyclonal serum fab with BG505 SOSIP.664:|
BG505 SOSIPv5.2 trimer / Polyclonal Fab
|Biological species||Human immunodeficiency virus 1 / Macaca mulatta (Rhesus monkey)|
|Method||single particle reconstruction / negative staining / Resolution: 26.5 Å|
|Authors||Nogal B / Ward AB|
|Funding support|| United States, 2 items |
|Citation||Journal: Cell Rep / Year: 2020|
Title: Mapping Polyclonal Antibody Responses in Non-human Primates Vaccinated with HIV Env Trimer Subunit Vaccines.
Authors: Bartek Nogal / Matteo Bianchi / Christopher A Cottrell / Robert N Kirchdoerfer / Leigh M Sewall / Hannah L Turner / Fangzhu Zhao / Devin Sok / Dennis R Burton / Lars Hangartner / Andrew B Ward /
Abstract: Rational immunogen design aims to focus antibody responses to vulnerable sites on primary antigens. Given the size of these antigens, there is, however, potential for eliciting unwanted, off-target ...Rational immunogen design aims to focus antibody responses to vulnerable sites on primary antigens. Given the size of these antigens, there is, however, potential for eliciting unwanted, off-target responses. Here, we use our electron microscopy polyclonal epitope mapping approach to describe the antibody specificities elicited by immunization of non-human primates with soluble HIV envelope trimers and subsequent repeated viral challenge. An increased diversity of epitopes recognized and the approach angle by which these antibodies bind constitute a hallmark of the humoral response in most protected animals. We also show that fusion peptide-specific antibodies are likely responsible for some neutralization breadth. Moreover, cryoelectron microscopy (cryo-EM) analysis of a fully protected animal reveals a high degree of clonality within a subset of putatively neutralizing antibodies, enabling a detailed molecular description of the antibody paratope. Our results provide important insights into the immune response against a vaccine candidate that entered into clinical trials in 2019.
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_20888.map.gz / Format: CCP4 / Size: 11.4 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 2.05 Å|
|Symmetry||Space group: 1|
CCP4 map header:
-Entire Polyclonal serum fab with BG505 SOSIP.664
|Entire||Name: Polyclonal serum fab with BG505 SOSIP.664 / Number of components: 3|
-Component #1: protein, Polyclonal serum fab with BG505 SOSIP.664
|Protein||Name: Polyclonal serum fab with BG505 SOSIP.664 / Recombinant expression: No|
-Component #2: protein, BG505 SOSIPv5.2 trimer
|Protein||Name: BG505 SOSIPv5.2 trimer / Recombinant expression: No|
|Source||Species: Human immunodeficiency virus 1|
|Source (engineered)||Expression System: unidentified (others)|
-Component #3: protein, Polyclonal Fab
|Protein||Name: Polyclonal Fab / Recombinant expression: No|
|Source||Species: Macaca mulatta (Rhesus monkey)|
|Specimen||Specimen state: Particle / Method: negative staining|
|Sample solution||pH: 4.5|
|Vitrification||Cryogen name: NONE|
-Electron microscopy imaging
Model: Tecnai Spirit / Image courtesy: FEI Company
|Imaging||Microscope: FEI TECNAI SPIRIT|
|Electron gun||Electron source: LAB6 / Accelerating voltage: 120 kV / Electron dose: 25 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Processing||Method: single particle reconstruction / Number of projections: 2500|
|3D reconstruction||Resolution: 26.5 Å / Resolution method: FSC 0.143 CUT-OFF|
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