Guillaume B. E. Stewart-Jones, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Paul V. Thomas, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Man Chen, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Aliaksandr Druz, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Gordon M. Joyce, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Wing-Pui Kong, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Mallika Sastry, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Conque Soto, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Yongping Yang, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Baoshan Zhang, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Lei Chen, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Gwo-Yu Chuang, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Ivelin S. Georgiev, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Jason S. McLellan, Dartmouth College

Document Type

Article

Publication Date

6-22-2015

Publication Title

PloS One

Department

Geisel School of Medicine

Abstract

Recombinant subunit vaccines should contain minimal non-pathogen motifs to reduce potential off-target reactivity. We recently developed a vaccine antigen against respiratory syncytial virus (RSV), which comprised the fusion (F) glycoprotein stabilized in its pre-fusion trimeric conformation by “DS-Cav1” mutations and by an appended C-terminal trimerization motif or “foldon” from T4-bacteriophage fibritin. Here we investigate the creation of a cyste- ine zipper to allow for the removal of the phage foldon, while maintaining the immunogenic- ity of the parent DS-Cav1+foldon antigen. Constructs without foldon yielded RSV F monomers, and enzymatic removal of the phage foldon from pre-fusion F trimers resulted in their dissociation into monomers. Because the native C terminus of the pre-fusion RSV F ectodomain encompasses a viral trimeric coiled-coil, we explored whether introduction of cysteine residues capable of forming inter-protomer disulfides might allow for stable trimers. Structural modeling indicated the introduced cysteines to form disulfide “rings”, with each ring comprising a different set of inward facing residues of the coiled-coil. Three sets of rings could be placed within the native RSV F coiled-coil, and additional rings could be added by duplicating portions of the coiled-coil. High levels of neutralizing activity in mice, equivalent to that of the parent DS-Cav1+foldon antigen, were elicited by a 4-ring stabilized RSV F trimer with no foldon. Structure-based alteration of a viral coiled-coil to create a cys- teine zipper thus allows a phage trimerization motif to be removed from a candidate vaccine antigen.

DOI

10.1371/journal.pone.0128779

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