The importance of RSV F protein conformation in VLPs in stimulation of neutralizing antibody titers in mice previously infected with RSV

Cullen, Lori McGinnes; Schmidt, Madelyn R.; Morrison, Trudy G.

doi:10.1080/21645515.2017.1329069

Cited by 31 publications

(65 citation statements)

References 34 publications

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“…It is also possible that RSV G interacts with RSV F on the virus surface, creating a quaternary structure that may limit RSV G to defined structures like those captured by the antibodies discussed in this paper. Interestingly, in an RSV virus-like particle vaccine containing F and G, the conformation of F affected the immunogenicity of G (59). These factors may be important in the design of an RSV vaccine.…”

Section: Discussionmentioning

confidence: 99%

Conformational Flexibility in Respiratory Syncytial Virus G Neutralizing Epitopes

Fedechkin

George

Castrejon

et al. 2020

J Virol

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Respiratory syncytial virus (RSV) is a top cause of severe lower respiratory tract disease and mortality in infants and the elderly. Currently, no vaccine or effective treatment exists for RSV. The RSV G glycoprotein mediates viral attachment to cells and contributes to pathogenesis by modulating host immunity through interactions with the human chemokine receptor CX3CR1. Antibodies targeting the RSV G central conserved domain are protective in both prophylactic and postinfection animal models. Here, we describe the crystal structure of the broadly neutralizing human monoclonal antibody 3G12 bound to the RSV G central conserved domain. Antibody 3G12 binds to a conformational epitope composed of highly conserved residues, explaining its broad neutralization activity. Surprisingly, RSV G complexed with 3G12 adopts a distinct conformation not observed in previously described RSV G-antibody structures. Comparison to other structures reveals that the RSV G central conserved domain is flexible and can adopt multiple conformations in the regions flanking the cysteine noose. We also show that restriction of RSV G flexibility with a proline mutation abolishes binding to antibody 3G12 but not antibody 3D3, which recognizes a different conformation of RSV G. Our studies provide new insights for rational vaccine design, indicating the importance of preserving both the global structural integrity of antigens and local conformational flexibility at antigenic sites, which may elicit a more diverse antibody response and broader protection against infection and disease. IMPORTANCE Respiratory syncytial virus (RSV) causes severe respiratory infections in infants, young children, and the elderly, and currently, no licensed vaccine exists. In this study, we describe the crystal structure of the RSV surface glycoprotein G in complex with a broadly neutralizing human monoclonal antibody. The antibody binds to RSV G at a highly conserved region stabilized by two disulfide bonds, but it captures RSV G in a conformation not previously observed, revealing that this region is both structured and flexible. Importantly, our findings provide insight for the design of vaccines that elicit diverse antibodies, which may provide broad protection from infection and disease. KEYWORDS X-ray crystallography, broadly neutralizing antibodies, protein structurefunction, respiratory syncytial virus R espiratory syncytial virus (RSV) is a globally prevalent virus that affects the airways and lungs. Infants and young children are at the highest risk of severe outcomes from RSV infection, with 33.1 million episodes of lower respiratory tract infection and approximately 3.2 million hospital visits and 118,200 deaths per year worldwide in children under the age of 5 years due to RSV (1). RSV is also a major cause of illness in adults older than 65 years of age and immunocompromised individuals, with an

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Section: Discussionmentioning

confidence: 99%

Conformational Flexibility in Respiratory Syncytial Virus G Neutralizing Epitopes

Fedechkin

George

Castrejon

et al. 2020

J Virol

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“…Using prefusion F-and postfusion F-containing VLPs, we assessed the influence of previous RSV infections on neutralizing antibody production in mice. We have reported that, in RSV-experienced mice, a single injection of a prefusion F-containing VLP stimulated high titers of neutralizing antibodies compared to those for postfusion F-containing VLPs or a second RSV infection (26). Here, we further characterized the immune response after VLP immunization of RSV-experienced mice by examining the durability of neutralizing antibody responses as well as levels, avidity, and durability of total anti-F protein IgG antibodies and the numbers of F protein-specific bone marrowassociated long-lived plasma cells and splenic memory B cells.…”

mentioning

confidence: 99%

Effect of Previous Respiratory Syncytial Virus Infection on Murine Immune Responses to F and G Protein-Containing Virus-Like Particles

Cullen

Schmidt

Morrison

2019

J Virol

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Most individuals are infected with respiratory syncytial virus (RSV) by age two, but infection does not result in long-term protective immunity to subsequent infections. Previous RSV infection may, however, impact responses to an RSV vaccine. The goal of these studies was to explore the effect of previous RSV infection on murine antibody responses to RSV F and G protein-containing virus-like particles (VLP), comparing responses to those resulting from VLP immunization of RSV-naive animals. These studies showed that after RSV infection, immunization with a single dose of VLPs containing a conformation-stabilized prefusion F protein stimulated high titers of neutralizing antibodies (NA), while an immunization with post-F-containing VLPs or a second RSV infection only weakly stimulated NA, even though total anti-F protein IgG antibody levels in both VLP-immunized animals were similar. Furthermore, single pre-F or post-F VLP immunization of animals previously infected (primed) with RSV resulted in total anti-F antibody titers that were 10- to 12-fold higher than titers after a VLP prime and boost of RSV-naive animals or after two consecutive RSV infections. The avidities of serum antibodies as well as numbers of splenic B cells and bone marrow cells after different immunization protocols were also assessed. The combined results show that RSV infection can quite effectively prime animals for the production of protective antibodies that can be efficiently activated by a pre-F VLP boost but not by a post-F VLP boost or a second RSV infection.IMPORTANCEHumans may experience repeated infections caused by the same serotype of respiratory syncytial virus (RSV), in contrast to infections with most other viruses, indicating that immune memory responses to RSV are defective. However, the effects of any residual but nonprotective immunity on responses to RSV vaccines are not clear. This study demonstrates that a VLP vaccine candidate containing a stabilized prefusion F protein can robustly stimulate protective immunity in animals previously infected with RSV, while a second RSV infection or a postfusion F-containing VLP cannot. This result shows that a properly constructed immunogen can be an effective vaccine in animals previously infected with RSV. The results also suggest that the defect in RSV memory is not in the induction of that memory but rather in its activation by a subsequent RSV infection.

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“…There is some debate over using either pre- or post-fusion F protein conformation. Several recent VLP approaches included the matrix (M) protein from RSV [33], HMPV [34], and NDV [36] and both pre- and post-fusion RSV F protein [33, 34, 36]. In all cases, the VLPs were immunogenic and triggered a desirable Th1 adaptive immune response [33, 34, 36].…”

Section: Virus-like Particles (Vlp)mentioning

confidence: 99%

“…Several recent VLP approaches included the matrix (M) protein from RSV [33], HMPV [34], and NDV [36] and both pre- and post-fusion RSV F protein [33, 34, 36]. In all cases, the VLPs were immunogenic and triggered a desirable Th1 adaptive immune response [33, 34, 36]. Although pre-fusion RSV F protein had a higher neutralizing antibody response than post-fusion RSV F protein, a combination of pre- and post-fusion F proteins produced the strongest Th1 response with the highest proportion of IgG2 antibodies [34].…”

Section: Virus-like Particles (Vlp)mentioning

confidence: 99%

Respiratory Syncytial Virus Vaccine Approaches: a Current Overview

Clark

Guerrero-Plata

2017

Curr Clin Micro Rpt

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Purpose of Review Respiratory syncytial virus (RSV) is a global human pathogen responsible for lower respiratory tract infections (LRTI). While RSV infection is innocuous in healthy adults, it is the leading cause of infant hospitalization for respiratory tract infection. Nearly everyone shows evidence of an RSV infection by the age of 3. However, there is still not a vaccine commercially available. This review will provide an update on the clinical and preclinical vaccine studies and different approaches to prevent RSV infection. Recent Findings Novel vaccine approaches that induce protection against RSV without enhancement of respiratory tract disease. Summary Recent technological approaches have led to generation of different strategies to prevent RSV infection, including live attenuated, chimeric, and subunit vaccines, virus-like particles, and nanoparticles. These vaccine approaches represent promising candidates towards an efficient RSV vaccine that effectively protects infants, children, and adults.

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The importance of RSV F protein conformation in VLPs in stimulation of neutralizing antibody titers in mice previously infected with RSV

Cited by 31 publications

References 34 publications

Conformational Flexibility in Respiratory Syncytial Virus G Neutralizing Epitopes

Conformational Flexibility in Respiratory Syncytial Virus G Neutralizing Epitopes

Effect of Previous Respiratory Syncytial Virus Infection on Murine Immune Responses to F and G Protein-Containing Virus-Like Particles

Respiratory Syncytial Virus Vaccine Approaches: a Current Overview

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