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Title: Potent single-domain antibodies that arrest respiratory syncytial virus fusion protein in its prefusion state

Authors:
; ; ; ; ; ORCiD logo; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
NIHFOREIGN
OSTI Identifier:
1347753
Resource Type:
Journal Article
Resource Relation:
Journal Name: Nature Communications; Journal Volume: 8; Journal Issue: 02, 2017
Country of Publication:
United States
Language:
ENGLISH

Citation Formats

Rossey, Iebe, Gilman, Morgan S. A., Kabeche, Stephanie C., Sedeyn, Koen, Wrapp, Daniel, Kanekiyo, Masaru, Chen, Man, Mas, Vicente, Spitaels, Jan, Melero, José A., Graham, Barney S., Schepens, Bert, McLellan, Jason S., and Saelens, Xavier. Potent single-domain antibodies that arrest respiratory syncytial virus fusion protein in its prefusion state. United States: N. p., 2017. Web. doi:10.1038/ncomms14158.
Rossey, Iebe, Gilman, Morgan S. A., Kabeche, Stephanie C., Sedeyn, Koen, Wrapp, Daniel, Kanekiyo, Masaru, Chen, Man, Mas, Vicente, Spitaels, Jan, Melero, José A., Graham, Barney S., Schepens, Bert, McLellan, Jason S., & Saelens, Xavier. Potent single-domain antibodies that arrest respiratory syncytial virus fusion protein in its prefusion state. United States. doi:10.1038/ncomms14158.
Rossey, Iebe, Gilman, Morgan S. A., Kabeche, Stephanie C., Sedeyn, Koen, Wrapp, Daniel, Kanekiyo, Masaru, Chen, Man, Mas, Vicente, Spitaels, Jan, Melero, José A., Graham, Barney S., Schepens, Bert, McLellan, Jason S., and Saelens, Xavier. Mon . "Potent single-domain antibodies that arrest respiratory syncytial virus fusion protein in its prefusion state". United States. doi:10.1038/ncomms14158.
@article{osti_1347753,
title = {Potent single-domain antibodies that arrest respiratory syncytial virus fusion protein in its prefusion state},
author = {Rossey, Iebe and Gilman, Morgan S. A. and Kabeche, Stephanie C. and Sedeyn, Koen and Wrapp, Daniel and Kanekiyo, Masaru and Chen, Man and Mas, Vicente and Spitaels, Jan and Melero, José A. and Graham, Barney S. and Schepens, Bert and McLellan, Jason S. and Saelens, Xavier},
abstractNote = {},
doi = {10.1038/ncomms14158},
journal = {Nature Communications},
number = 02, 2017,
volume = 8,
place = {United States},
year = {Mon Feb 13 00:00:00 EST 2017},
month = {Mon Feb 13 00:00:00 EST 2017}
}
  • Human respiratory syncytial virus (hRSV) has two major surface glycoproteins (G and F) anchored in the lipid envelope. Membrane fusion promoted by hRSV{sub F} occurs via refolding from a pre-fusion form to a highly stable post-fusion state involving large conformational changes of the F trimer. One of these changes results in assembly of two heptad repeat sequences (HRA and HRB) into a six-helix bundle (6HB) motif. To assist in distinguishing pre- and post-fusion conformations of hRSV{sub F}, we have prepared polyclonal (α-6HB) and monoclonal (R145) rabbit antibodies specific for the 6HB. Among other applications, these antibodies were used to exploremore » the requirements of 6HB formation by isolated protein segments or peptides and by truncated mutants of the F protein. Site-directed mutagenesis and electron microscopy located the R145 epitope in the post-fusion hRSV{sub F} at a site distantly located from previously mapped epitopes, extending the repertoire of antibodies that can decorate the F molecule. - Highlights: • Antibodies specific for post-fusion respiratory syncytial virus fusion protein are described. • Polyclonal antibodies were obtained in rabbit inoculated with chimeric heptad repeats. • Antibody binding required assembly of a six-helix bundle in the post-fusion protein. • A monoclonal antibody with similar structural requirements is also described. • Binding of this antibody to the post-fusion protein was visualized by electron microscopy.« less
  • A safe and effective vaccine against respiratory syncytial virus (RSV) should confer protection without causing vaccine-enhanced disease. Here, using a cotton rat model, we investigated the protective efficacy and safety of an RSV combination vaccine composed of F-encoding plasmid DNA and virus-like particles containing RSV fusion (F) and attachment (G) glycoproteins (FFG-VLP). Cotton rats with FFG-VLP vaccination controlled lung viral replication below the detection limit, and effectively induced neutralizing activity and antibody-secreting cell responses. In comparison with formalin inactivated RSV (FI-RSV) causing severe RSV disease after challenge, FFG-VLP vaccination did not cause weight loss, airway hyper-responsiveness, IL-4 cytokines, histopathology, andmore » infiltrates of proinflammatory cells such as eosinophils. FFG-VLP was even more effective in preventing RSV-induced pulmonary inflammation than live RSV infections. This study provides evidence that FFG-VLP can be developed into a safe and effective RSV vaccine candidate. - Highlights: • Combined RSV FFG VLP vaccine is effective in inducing F specific responses. • FFG VLP vaccine confers RSV neutralizing activity and viral control in cotton rats. • Cotton rats with RSV FFG VLP vaccination do not show vaccine-enhanced disease. • Cotton rats with FFG VLP vaccine induce F specific antibody secreting cell responses. • Cotton rats with FFG VLP do not induce lung cellular infiltrates and Th2 cytokine.« less
  • The role of binding kinetics in determining neutralizing potency for antiviral antibodies is poorly understood. While it is believed that increased steady-state affinity correlates positively with increased virus-neutralizing activity, the relationship between association or dissociation rate and neutralization potency is unclear. We investigated the effect of naturally-occurring antibody resistance mutations in the RSV F protein on the kinetics of binding to palivizumab. Escape from palivizumab-mediated neutralization of RSV occurred with reduced association rate (K{sub on}) for binding to RSV F protein, while alteration of dissociation rate (K{sub off}) did not significantly affect neutralizing activity. Interestingly, linkage of reduced K{sub on}more » with reduced potency mirrored the effect of increased K{sub on} found in a high-affinity enhanced potency palivizumab variant (motavizumab). These data suggest that association rate is the dominant factor driving neutralization potency for antibodies to RSV F protein antigenic site A and determines the potency of antibody somatic variants or efficiency of escape of viral glycoprotein variants. - Highlights: • The relationship of affinity to neutralization for virus antibodies is uncertain. • Palivizumab binds to RSV escape mutant fusion proteins, but with reduced affinity. • Association rate (K{sub on}) correlated well with the potency of neutralization.« less
  • Paramyxoviruses have been shown to produce proteins that inhibit interferon production and signaling. For human respiratory syncytial virus (RSV), the nonstructural NS1 and NS2 proteins have been shown to have interferon antagonist activity through an unknown mechanism. To understand further the functions of NS1 and NS2, we generated recombinant RSV in which both NS1 and NS2 were replaced by the PIV5 V protein, which has well-characterized IFN antagonist activities ({delta}NS1/2-V). Expression of V was able to partially inhibit IFN responses in {delta}NS1/2-V-infected cells. In addition, the replication kinetics of {delta}NS1/2-V were intermediate between {delta}NS1/2 and wild-type (rA2) in A549 cells.more » However, expression of V did not affect the ability of {delta}NS1/2-V to activate IRF3 nuclear translocation and IFN{beta} transcription. These data indicate that V was able to replace some of the IFN inhibitory functions of the RSV NS1 and NS2 proteins, but also that NS1 and NS2 have functions in viral replication beyond IFN antagonism.« less