Structure of the prefusion-locking broadly neutralizing antibody RVC20 bound to the rabies virus glycoprotein

Hellert, Jan; Buchrieser, Julian; Larrous, Florence; Minola, Andrea; Melo, Guilherme Dias de; Soriaga, Leah; England, Patrick; Haouz, Ahmed; Telenti, Amalio; Corti, Davide; Bourhy, Hervé; Rey, F.A.

doi:10.1038/s41467-020-14398-7

Cited by 32 publications

(50 citation statements)

References 29 publications

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“…In the quest for a novel therapeutic possibility, we have previously reported the selection of two human monoclonal antibodies (mAbs), RVC20 and RVC58, that were able to bind to two distinct antigenic sites on the RABV glycoprotein protein (sites I and III), to potently neutralize RABV isolates of all lineages, and of all phylogroup I non‐RABV isolates, and that presented a protective role when used as early PEP in hamsters as well (De Benedictis et al , 2016; Hellert et al , 2020).…”

Section: Introductionmentioning

confidence: 99%

A combination of two human monoclonal antibodies cures symptomatic rabies

et al. 2020

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Rabies is a neglected disease caused by a neurotropic Lyssavirus, transmitted to humans predominantly by the bite of infected dogs. Rabies is preventable with vaccines or proper post‐exposure prophylaxis (PEP), but it still causes about 60,000 deaths every year. No cure exists after the onset of clinical signs, and the case‐fatality rate approaches 100% even with advanced supportive care. Here, we report that a combination of two potent neutralizing human monoclonal antibodies directed against the viral envelope glycoprotein cures symptomatic rabid mice. Treatment efficacy requires the concomitant administration of antibodies in the periphery and in the central nervous system through intracerebroventricular infusion. After such treatment, recovered mice presented good clinical condition, viral loads were undetectable, and the brain inflammatory profile was almost normal. Our findings provide the unprecedented proof of concept of an antibody‐based therapeutic approach for symptomatic rabies.

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Section: Introductionmentioning

confidence: 99%

A combination of two human monoclonal antibodies cures symptomatic rabies

et al. 2020

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“…Recent studies have confirmed the crystal structures of RABV G and its interaction with neutralizing antibodies (Hellert et al, 2020;Yang et al, 2020). Residues from G333-G350 of ectodomain from three strains (CVS-11, Flury, and SAD-B19) are observed to bond to antibody 523-11 (Yang et al, 2020).…”

Section: Figure 5 | (A)mentioning

confidence: 76%

Amino Acid Mutation in Position 349 of Glycoprotein Affect the Pathogenicity of Rabies Virus

Luo

Zhang

et al. 2020

Front. Microbiol.

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Rabies, caused by rabies virus (RABV), is a zoonotic disease infecting mammals including humans. Studies have confirmed that glycoprotein (G) is most related to RABV pathogenicity. In the present study, to discover more amino acid sites related to viral pathogenicity, artificial mutants have been constructed in G of virulent strain GD-SH-01 backbone. Results showed that pathogenicity of GD-SH-01 significantly decreased when Gly 349 was replaced by Glu 349 through in vivo assays. Gly 349 →Glu 349 of G did not significantly influence viral growth and spread in NA cells. Gly 349 →Glu 349 of G increased the immunogenicity of GD-SH-01 in periphery and induced more expression of interferon alpha (IFN-α) in the brain in mice. It was observed that Gly 349 →Glu 349 of G led to enhanced blood-brain barrier (BBB) permeability at day 5 postinfection. All together, these data revealed that Gly 349 →Glu 349 of G mutation decreased RABV pathogenicity through enhanced immune response and increased BBB permeability. This study provides a new referenced site G349 that could attenuate pathogenicity of RABV.

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“…Such a mechanism would explain the disparity in neutralizing activity between mAbs 93k and SG2, with the latter not binding to either the N-terminus or β30, which our previous study and this study demonstrate are essential regions for gB fusion function. There have been several studies demonstrating that mAbs can prevent viral fusion proteins from undergoing conformational changes [50-52]. A single study to date provides some evidence that a similar phenomenon is plausible for herpesvirus gB [53].…”

Section: Discussionmentioning

confidence: 99%

The N-terminus of varicella-zoster virus glycoprotein B has a functional role in fusion

Oliver

Xing²,

Chen

et al. 2020

Preprint

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Varicella-zoster virus (VZV) is a medically important alphaherpesvirus that induces fusion of the virion envelope and the cell membrane during entry, and between cells to form polykaryocytes within infected tissues during pathogenesis. All members of the Herpesviridae, including VZV, have a conserved core fusion complex composed of glycoproteins, gB, gH and gL. The ectodomain of the primary fusogen, gB, has five domains, DI-V, of which DI contains the fusion loops needed for fusion function. We recently demonstrated that DIV is critical for fusion initiation, which was revealed by a 2.8Å structure of a VZV neutralizing mAb, 93k, bound to gB and mutagenesis of the gB-93k interface. To further assess the mechanism of mAb 93k neutralization, the binding site of a non-neutralizing mAb to gB, SG2, was compared to mAb 93k using single particle cryogenic electron microscopy (cryo-EM). The gB-SG2 interface partially overlapped with that of gB-93k but, unlike mAb 93k, mAb SG2 did not interact with the gB N-terminus, suggesting a potential role for the gB N-terminus in membrane fusion. The gB ectodomain structure in the absence of antibody was defined at near atomic resolution by single particle cryo-EM (3.9Å) of native full-length gB purified from infected cells and by X-ray crystallography (2.4Å) of the transiently expressed ectodomain. Both structures revealed that the VZV gB N-terminus (aa72-114) was flexible based on the absence of visible structures in the cryo-EM or X-ray crystallography data but the presence of gB N-terminal peptides were confirmed by mass spectrometry. Notably, N-terminal residues 109KSQD112 were predicted to form a small α-helix and alanine substitution of these residues abolished cell-cell fusion in a virus-free assay. Importantly, transferring the 109AAAA112 mutation into the VZV genome significantly impaired viral propagation. These data establish a functional role for the gB N-terminus in membrane fusion broadly relevant to the Herpesviridae.Author SummaryHerpesviruses are ubiquitous infectious agents of medical and economic importance, including varicella-zoster virus (VZV), which causes chicken pox and shingles. A unifying theme of herpesviruses is their mechanism of entry into host cells, membrane fusion, via a core complex of virally expressed envelope glycoproteins gB, gH and gL. Of these, the primary fusogen, gB, is activated by the heterodimer gH-gL through an unknown mechanism and enables the virus envelope to merge with cell membranes to release the DNA containing capsid into the cytoplasm to initiate infection. By using a human antibody that neutralizes VZV we have recently demonstrated that the initiation of membrane fusion is associated with the crown region of gB. Here, we use cryogenic electron microscopy to compare the structure of this human neutralizing antibody, 93k, to a non-neutralizing antibody SG2. Surprisingly, both antibodies bind to the crown of gB with considerable overlap of their footprints on gB with one important exception, SG2 does not bind to a flexible region in the gB N-terminus. Mutations incorporated into this flexible region disrupts gB mediated membrane fusion and significantly impairs VZV propagation, identifying an Achilles heel in viral replication.

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Structure of the prefusion-locking broadly neutralizing antibody RVC20 bound to the rabies virus glycoprotein

Cited by 32 publications

References 29 publications

A combination of two human monoclonal antibodies cures symptomatic rabies

A combination of two human monoclonal antibodies cures symptomatic rabies

Amino Acid Mutation in Position 349 of Glycoprotein Affect the Pathogenicity of Rabies Virus

The N-terminus of varicella-zoster virus glycoprotein B has a functional role in fusion

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