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- EMDB-24861: Modified BG505 SOSIP-based immunogen RC1 in complex with NHP 1 po... -

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Basic information

Entry
Database: EMDB / ID: EMD-24861
TitleModified BG505 SOSIP-based immunogen RC1 in complex with NHP 1 post-Boost 1 polyclonal Fabs
Map dataSharpened map volume
Sample
  • Complex: Modified BG505 SOSIP-based immunogen RC1 in complex with NHP 1 post-Boost 1 polyclonal Fabs
    • Complex: Modified BG505 SOSIP-based immunogen RC1
    • Complex: NHP 1 post-Boost 1 polyclonal Fabs
Biological speciesHuman immunodeficiency virus 1 / Macaca mulatta (Rhesus monkey)
Methodsingle particle reconstruction / negative staining / Resolution: 17.0 Å
AuthorsDeLaitsch AT / Barnes CO / Bjorkman PJ
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)P01-AI100148 United States
CitationJournal: Sci Transl Med / Year: 2021
Title: Sequential immunization of macaques elicits heterologous neutralizing antibodies targeting the V3-glycan patch of HIV-1 Env.
Authors: Amelia Escolano / Harry B Gristick / Rajeev Gautam / Andrew T DeLaitsch / Morgan E Abernathy / Zhi Yang / Haoqing Wang / Magnus A G Hoffmann / Yoshiaki Nishimura / Zijun Wang / Nicholas ...Authors: Amelia Escolano / Harry B Gristick / Rajeev Gautam / Andrew T DeLaitsch / Morgan E Abernathy / Zhi Yang / Haoqing Wang / Magnus A G Hoffmann / Yoshiaki Nishimura / Zijun Wang / Nicholas Koranda / Leesa M Kakutani / Han Gao / Priyanthi N P Gnanapragasam / Henna Raina / Ana Gazumyan / Melissa Cipolla / Thiago Y Oliveira / Victor Ramos / Darrell J Irvine / Murillo Silva / Anthony P West / Jennifer R Keeffe / Christopher O Barnes / Michael S Seaman / Michel C Nussenzweig / Malcolm A Martin / Pamela J Bjorkman /
Abstract: Broadly neutralizing antibodies (bNAbs) against HIV-1 develop after prolonged virus and antibody coevolution. Previous studies showed that sequential immunization with a V3-glycan patch germline- ...Broadly neutralizing antibodies (bNAbs) against HIV-1 develop after prolonged virus and antibody coevolution. Previous studies showed that sequential immunization with a V3-glycan patch germline-targeting HIV-1 envelope trimer (Env) followed by variant Envs can reproduce this process in mice carrying V3-glycan bNAb precursor B cells. However, eliciting bNAbs in animals with polyclonal antibody repertoires is more difficult. We used a V3-glycan immunogen multimerized on virus-like particles (VLPs), followed by boosting with increasingly native-like Env-VLPs, to elicit heterologous neutralizing antibodies in nonhuman primates (NHPs). Structures of antibody/Env complexes after prime and boost vaccinations demonstrated target epitope recognition with apparent maturation to accommodate glycans. However, we also observed increasing off-target antibodies with boosting. Eight vaccinated NHPs were subsequently challenged with simian-human immunodeficiency virus (SHIV), and seven of eight animals became infected. The single NHP that remained uninfected after viral challenge exhibited one of the lowest neutralization titers against the challenge virus. These results demonstrate that more potent heterologous neutralization resulting from sequential immunization is necessary for protection in this animal model. Thus, improved prime-boost regimens to increase bNAb potency and stimulate other immune protection mechanisms are essential for developing anti–HIV-1 vaccines.
History
DepositionSep 13, 2021-
Header (metadata) releaseApr 6, 2022-
Map releaseApr 6, 2022-
UpdateApr 6, 2022-
Current statusApr 6, 2022Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_24861.png

Downloads & links

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Map

FileDownload / File: emd_24861.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationSharpened map volume
Voxel sizeX=Y=Z: 1.44 Å
Density
Contour LevelBy AUTHOR: 0.1
Minimum - Maximum-0.19442385 - 0.46670094
Average (Standard dev.)-0.003147106 (±0.0313359)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions256256256
Spacing256256256
CellA=B=C: 368.64 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Modified BG505 SOSIP-based immunogen RC1 in complex with NHP 1 po...

EntireName: Modified BG505 SOSIP-based immunogen RC1 in complex with NHP 1 post-Boost 1 polyclonal Fabs
Components
  • Complex: Modified BG505 SOSIP-based immunogen RC1 in complex with NHP 1 post-Boost 1 polyclonal Fabs
    • Complex: Modified BG505 SOSIP-based immunogen RC1
    • Complex: NHP 1 post-Boost 1 polyclonal Fabs

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Supramolecule #1: Modified BG505 SOSIP-based immunogen RC1 in complex with NHP 1 po...

SupramoleculeName: Modified BG505 SOSIP-based immunogen RC1 in complex with NHP 1 post-Boost 1 polyclonal Fabs
type: complex / ID: 1 / Parent: 0

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Supramolecule #2: Modified BG505 SOSIP-based immunogen RC1

SupramoleculeName: Modified BG505 SOSIP-based immunogen RC1 / type: complex / ID: 2 / Parent: 1
Source (natural)Organism: Human immunodeficiency virus 1
Recombinant expressionOrganism: Homo sapiens (human)

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Supramolecule #3: NHP 1 post-Boost 1 polyclonal Fabs

SupramoleculeName: NHP 1 post-Boost 1 polyclonal Fabs / type: complex / ID: 3 / Parent: 1
Source (natural)Organism: Macaca mulatta (Rhesus monkey)

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Experimental details

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Structure determination

Methodnegative staining
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 8
Component:
ConcentrationNameFormula
0.02 MTris
0.15 MSodium ChlorideNaClSodium chloride
StainingType: NEGATIVE / Material: Uranyl Formate / Details: 1.5% Uranyl Formate
Detailsmonodisperse sample

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Electron microscopy

MicroscopeFEI TECNAI ARCTICA
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal magnification: 28000
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 10.0 e/Å2
Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company

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Image processing

Startup modelType of model: OTHER / Details: ab initio
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
Final reconstructionApplied symmetry - Point group: C3 (3 fold cyclic) / Resolution.type: BY AUTHOR / Resolution: 17.0 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 2.14) / Number images used: 55890

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