Structure of trimeric pre-fusion rabies virus glycoprotein in complex with two protective antibodies

Ng, Weng M; Fedosyuk, Sofiya; S, English; Augusto, Gilles; Berg, Adam; Thorley, Luke; Haselon, Anna-Sophie; Segireddy, Rameswara R; Bowden, Thomas A.; Douglas, Alexander D.

doi:10.1016/j.chom.2022.07.014

Cited by 23 publications

(25 citation statements)

References 65 publications

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“…To map sites of vulnerability to neutralization on RABV-G, we analyzed mAb epitopes on the spike (Fig 3). To date, five structures of neutralizing antibodies in complex with RABV-G have been reported, of which mAb 17C7 [16], mAb 523-11 [18], and mAb RVA122 [15] target epitopes located in domains I and II, whereas mAb RVC20 [33] and mAb 1112-1 target domain III [16]. In addition to these recent structural insights, earlier competition ELISA assays have outlined six antigenic sites (sites i-iv, minor site ‘a’ and G5) on RABV-G [22–24, 34–36] which are located in domains I to III as described in Table S2.…”

Section: Resultsmentioning

confidence: 99%

“…We note that in line with the abrogation of receptor binding as one of the mechanisms of virus neutralization, two regions implicated for receptor binding overlap with recently identified epitopes of neutralizing antibodies. Firstly, the nicotinic acetylcholine receptor (nAChR)-binding peptide at domain III residues 175-203 [16, 37, 38] falls within the epitopes of mAbs CR57 and RVC20, as discussed in more detail below. Secondly, mAbs 62-7-13 [11], RVA122 [15], 17C7 [16], CR4098 [22, 23], and 523-11[18] bind proximal to putative p75 neurotrophin receptor (p75NTR)-engaging residues K330 and R333 in domain I [39].…”

Section: Resultsmentioning

confidence: 99%

“…The bullet-shaped RABV nucleocapsid is enclosed by a lipid bilayer envelope that displays trimeric G spikes, which mediate host cell entry and receptor recognition [2,3,13], containing an ectodomain that displays the classical rhabdoviral class III fusion protein fold [13][14][15][16][17][18]. The ectodomain is subdivided into four distinct domains: the central domain, which encompasses domains I and II, domain III (referred to as the pleckstrin homology domain), and domain IV (known as the fusion domain) [15,16,18]. RABV-G initiates infection by binding to host-cell receptors, followed by viral uptake by endocytosis.…”

Section: Introductionmentioning

confidence: 99%

“…This conformational transition of the spike from a prefusion state (at neutral pH) to a postfusion state (at low pH) is facilitated by hinge regions located between domains II and III, and domains III and IV [14, 15, 21]. While the structures of the intermediary steps between pre- and postfusion states remain unknown, recent insights into the RABV-G prefusion spike in complex with neutralizing antibodies are beginning to highlight key antigenic regions on RABV-G [15, 16]. Further, there exists an extensive array of neutralizing antibodies that have yet to be structurally characterized [22, 23] and which comprise prime targets for studies aimed at identifying functionally critical spike regions.…”

Section: Introductionmentioning

confidence: 99%

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Structural insight into rabies virus neutralization revealed by an engineered antibody scaffold

Kedari,

Iheozor-Ejiofor,

Levanov

et al. 2023

Preprint

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Host-cell entry of the highly pathogenic rabies virus (RABV) is mediated by trimeric glycoprotein (G) spikes, which also represent the primary target for the humoral immune response. RABV-G displays several antigenic sites targeted by neutralizing antibodies, including monoclonal antibodies (mAbs) which have been proposed as quality-controlled alternatives to traditional polyclonal rabies immunoglobulin treatment. In this study, we determine the epitope of a potently neutralizing human anti-rabies mAb, CR57, which we engineered into a diabody to facilitate crystallization. We report the crystal structure of the CR57 diabody alone at 2.38 Å resolution, and in complex with RABV-G domain III at 3.15 Å resolution. CR57 is demonstrated to bind RABV through a predominantly hydrophobic interface, with essential interactions targeting a conserved six-residue peptide sequence ’KLCGVL’ on the RABV-G. Further, our structural analysis suggests that CR57 sterically hinders receptor recognition and the fusogenic transitions of the spike protein that are required for host-cell entry. Altogether, this investigation provides a structural perspective on rabies inhibition by a potent antibody and delineates a functionally significant region in the spike. This understanding could pave the way for the development of prophylactic antibodies and other therapeutic strategies.Author summaryRabies virus (RABV) and many other lyssaviruses possess the ability to invade the central nervous system, leading to fatal encephalitis in mammals. Initiation of the infectious cycle depends on host cell recognition and entry, which is mediated by viral surface glycoprotein (G) spikes and can be inhibited by spike-targeting neutralizing antibodies. In our study, we elucidated the crystal structure of an antigenic domain from RABV-G in complex with a diabody derived from the potently neutralizing antibody CR57. This investigation revealed the molecular interactions by which CR57 binds to RABV-G and outlined a site of vulnerability comprising a conserved peptide in RABV-G domain III, where antibody binding is likely to inhibit RABV by obstructing host cell entry. Insights into the binding modalities of antibodies like CR57 deepen our understanding of how RABV and other lyssaviruses are neutralized, aiding the development of potential therapeutics. Furthermore, our study showcases the utility of engineering antibodies into diabodies to obtain crystal structures of antibody-antigen complexes.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Structural insight into rabies virus neutralization revealed by an engineered antibody scaffold

Kedari,

Iheozor-Ejiofor,

Levanov

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…In this proof-of-concept study with a limited number of animals, an immune response was detected in some but not all animals, clearly indicating the need for improvements if the requirements for registrations by EMA (58) and WOAH (61) are to be met. Potential parameters for an improved immune response are genetic modifications to the gene insert to increase its stability (62). Also, in this study, a dose of 10 8.5 TCID 50 /animal was used.…”

Section: Discussionmentioning

confidence: 99%

Oral immunization of goats and foxes with a recombinant NDV vectored rabies vaccine

Murr,

Freuling,

Pérez-Bravo

et al. 2023

Preprint

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Vaccination of the reservoir species is a key component in the global fight against rabies. For wildlife reservoir species and hard to reach spillover species (e. g. ruminant farm animals), oral vaccination is the only solution. In search for a novel potent and safe oral rabies vaccine, we generated a recombinant vector virus based on lentogenic Newcastle disease virus (NDV) strain Clone 30 that expresses the glycoprotein G of rabies virus (RABV) vaccine strain SAD L16 (rNDV_GRABV). Transgene expression and virus replication was verified in avian and mammalian cells. To test immunogenicity and viral shedding, in a proof-of-concept study six goats and foxes, representing herbivore and carnivore species susceptible to rabies, each received a single dose of rNDV_GRABV (108.5 TCID50/animal) by direct oral application. For comparison, three animals received the similar dose of the empty viral vector (rNDV). All animals remained clinically inconspicuous during the trial. Viral RNA could be isolated from oral and nasal swabs until four (goats) or seven days (foxes) post vaccination, while infectious NDV could not be re-isolated. After four weeks, three out of six rNDV_GRABV vaccinated foxes developed RABV binding and virus neutralizing antibodies. Five out of six rNDV_GRABV vaccinated goats displayed RABV G specific antibodies either detected by ELISA or RFFIT. Additionally, NDV and RABV specific T cell activity was demonstrated in some of the vaccinated animals by detecting antigen specific interferon γ secretion in lymphocytes isolated from pharyngeal lymph nodes. In conclusion, the NDV vectored rabies vaccine rNDV_GRABV was safe and immunogenic after a single oral application in goats and foxes, and highlight the potential of NDV as vector for oral vaccines  in mammals.

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A Cholesterol Metabolic Regulated Hydrogen‐Bonded Organic Framework (HOF)‐Based Biotuner for Antibody Non‐Dependent Immunotherapy Tailored for Glioblastoma

et al. 2023

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The metabolic reprogramming of glioblastoma (GBM) poses a tremendous obstacle to effective immunotherapy due to its impact on the immunosuppressive microenvironment. Here, we develop a hydrogen‐bonded organic frameworks (HOF) specifically designed for GBM immunotherapy, taking advantage of the relatively isolated cholesterol metabolism microenvironment in the central nervous system (CNS). The HOF‐based biotuner regulates extra/intracellular cholesterol metabolism, effectively blocking the programmed cell death protein 1/programmed death‐ligand 1 (PD‐1/PD‐L1) pathway and reducing 2B4 expression. This metabolically disrupts the immunosuppressive microenvironment of GBM and rejuvenates CD8+ T cells. Moreover, cholesterol metabolism regulation offers additional benefits in treating GBM invasion. Furthermore, in response to the tumor microenvironment (TEM)‐initiated chemiexcited photodynamic therapy (PDT), which was enhanced during the regulation of cholesterol metabolism, the biotuner could effectively trigger immunogenic cell death (ICD) and increase the infiltration of cytotoxic T lymphocytes (CTLs) in GBM. By reversing immunosuppressive microenvironment and bolstering chemiexcited‐PDT, our approach invigorates efficient antibody non‐dependent immunotherapy for GBM. This study provides a model for enhancing immunotherapy through cholesterol metabolism regulation and explores the feasibility of a “metabolic checkpoint” strategy in GBM treatment.This article is protected by copyright. All rights reserved

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Structure of trimeric pre-fusion rabies virus glycoprotein in complex with two protective antibodies

Cited by 23 publications

References 65 publications

Structural insight into rabies virus neutralization revealed by an engineered antibody scaffold

Structural insight into rabies virus neutralization revealed by an engineered antibody scaffold

Oral immunization of goats and foxes with a recombinant NDV vectored rabies vaccine

A Cholesterol Metabolic Regulated Hydrogen‐Bonded Organic Framework (HOF)‐Based Biotuner for Antibody Non‐Dependent Immunotherapy Tailored for Glioblastoma

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