|Entry||Database: EMDB / ID: EMD-22715|
|Title||nsEM map of the 16055 SOSIP HIV Env trimer in complex with polyclonal Fab from rabbit r2464|
|Sample||nsEM map of the 16055 SOSIP HIV Env trimer in complex with polyclonal Fab from rabbit r2464 (Week 22)|
|Biological species||synthetic construct (others)|
|Method||single particle reconstruction / negative staining / Resolution: 15.9 Å|
|Authors||Ward AB / Antanasijevic A|
|Funding support|| United States, 1 items |
|Citation||Journal: NPJ Vaccines / Year: 2021|
Title: Immunofocusing and enhancing autologous Tier-2 HIV-1 neutralization by displaying Env trimers on two-component protein nanoparticles.
Authors: Philip J M Brouwer / Aleksandar Antanasijevic / Marlon de Gast / Joel D Allen / Tom P L Bijl / Anila Yasmeen / Rashmi Ravichandran / Judith A Burger / Gabriel Ozorowski / Jonathan L Torres / ...Authors: Philip J M Brouwer / Aleksandar Antanasijevic / Marlon de Gast / Joel D Allen / Tom P L Bijl / Anila Yasmeen / Rashmi Ravichandran / Judith A Burger / Gabriel Ozorowski / Jonathan L Torres / Celia LaBranche / David C Montefiori / Rajesh P Ringe / Marit J van Gils / John P Moore / Per Johan Klasse / Max Crispin / Neil P King / Andrew B Ward / Rogier W Sanders /
Abstract: The HIV-1 envelope glycoprotein trimer is poorly immunogenic because it is covered by a dense glycan shield. As a result, recombinant Env glycoproteins generally elicit inadequate antibody levels ...The HIV-1 envelope glycoprotein trimer is poorly immunogenic because it is covered by a dense glycan shield. As a result, recombinant Env glycoproteins generally elicit inadequate antibody levels that neutralize clinically relevant, neutralization-resistant (Tier-2) HIV-1 strains. Multivalent antigen presentation on nanoparticles is an established strategy to increase vaccine-driven immune responses. However, due to nanoparticle instability in vivo, the display of non-native Env structures, and the inaccessibility of many neutralizing antibody (NAb) epitopes, the effects of nanoparticle display are generally modest for Env trimers. Here, we generate two-component self-assembling protein nanoparticles presenting twenty SOSIP trimers of the clade C Tier-2 genotype 16055. We show in a rabbit immunization study that these nanoparticles induce 60-fold higher autologous Tier-2 NAb titers than the corresponding SOSIP trimers. Epitope mapping studies reveal that the presentation of 16055 SOSIP trimers on these nanoparticle focuses antibody responses to an immunodominant apical epitope. Thus, these nanoparticles are a promising platform to improve the immunogenicity of Env trimers with apex-proximate NAb epitopes.
|Validation Report||Summary, Full report, XML, About validation report|
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_22715.map.gz / Format: CCP4 / Size: 42.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 1.77 Å|
|Symmetry||Space group: 1|
CCP4 map header:
-Entire nsEM map of the 16055 SOSIP HIV Env trimer in complex with polycl...
|Entire||Name: nsEM map of the 16055 SOSIP HIV Env trimer in complex with polyclonal Fab from rabbit r2464 (Week 22)|
Number of components: 1
-Component #1: protein, nsEM map of the 16055 SOSIP HIV Env trimer in complex wi...
|Protein||Name: nsEM map of the 16055 SOSIP HIV Env trimer in complex with polyclonal Fab from rabbit r2464 (Week 22)|
Recombinant expression: No
|Source||Species: synthetic construct (others)|
|Source (engineered)||Expression System: Homo sapiens (human) / Vector: pPPI4 / Strain: HEK293F|
|Specimen||Specimen state: Particle / Method: negative staining|
|Sample solution||Specimen conc.: 0.05 mg/mL / Buffer solution: TBS buffer, pH 7.4 / pH: 7.4|
|Staining||Sample diluted to 0.05 mg/mL. 3 uL was applied onto the grid, blotted off, and then stained with 2% uranyl formate for 60 seconds.|
|Vitrification||Cryogen name: NONE|
-Electron microscopy imaging
Model: Tecnai F20 / Image courtesy: FEI Company
|Imaging||Microscope: FEI TECNAI F20|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Electron dose: 25 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Magnification: 62000 X (nominal) / Cs: 2.7 mm / Imaging mode: BRIGHT FIELD / Defocus: 1500.0 nm|
|Specimen Holder||Model: OTHER|
|Camera||Detector: TVIPS TEMCAM-F416 (4k x 4k)|
|Processing||Method: single particle reconstruction / Applied symmetry: C1 (asymmetric) / Number of projections: 69539|
|3D reconstruction||Algorithm: BACK PROJECTION / Software: RELION / Resolution: 15.9 Å / Resolution method: FSC 0.143 CUT-OFF|
-Atomic model buiding
|Modeling #1||Details: No fitting was performed|
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