title: Sensitivity of immune response quality to influenza helix 190 antigen structure displayed on a modular virus-like particle.
abstract: Biomolecular engineering enables synthesis of improved proteins through synergistic fusion of modules from unrelated biomolecules. Modularization of peptide antigen from an unrelated pathogen for presentation on a modular virus-like particle (VLP) represents a new and promising approach to synthesize safe and efficacious vaccines. Addressing a key knowledge gap in modular VLP engineering, this study investigates the underlying fundamentals affecting the ability of induced antibodies to recognize the native pathogen. Specifically, this quality of immune response is correlated to the peptide antigen module structure. We modularized a helical peptide antigen element, helix 190 (H190) from the influenza hemagglutinin (HA) receptor binding region, for presentation on murine polyomavirus VLP, using two strategies aimed to promote H190 helicity on the VLP. In the first strategy, H190 was flanked by GCN4 structure-promoting elements within the antigen module; in the second, dual H190 copies were arrayed as tandem repeats in the module. Molecular dynamics simulation predicted that tandem repeat arraying would minimize secondary structural deviation of modularized H190 from its native conformation. In vivo testing supported this finding, showing that although both modularization strategies conferred high H190-specific immunogenicity, tandem repeat arraying of H190 led to a strikingly higher immune response quality, as measured by ability to generate antibodies recognizing a recombinant HA domain and split influenza virion. These findings provide new insights into the rational engineering of VLP vaccines, and could ultimately enable safe and efficacious vaccine design as an alternative to conventional approaches necessitating pathogen cultivation.
authors.affiliation: University of Queensland, Australian Institute for Bioengineering and Nanotechnology, Centre for Biomolecular Engineering, St. Lucia, QLD 4072, Australia.
authors.forename: Melisa R
authors.initials: MR
authors.lastname: Anggraeni
authors.forename: Natalie K
authors.initials: NK
authors.lastname: Connors
authors.forename: Yang
authors.initials: Y
authors.lastname: Wu
authors.forename: Yap P
authors.initials: YP
authors.lastname: Chuan
authors.forename: Linda H L
authors.initials: LH
authors.lastname: Lua
authors.forename: Anton P J
authors.initials: AP
authors.lastname: Middelberg
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D005260:Female; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000276:immunology*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology; D007252:Influenza Vaccines / Q000276:immunology*; D007251:Influenza, Human / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D056004:Molecular Dynamics Simulation; D009976:Orthomyxoviridae Infections / Q000276:immunology; D011120:Polyomavirus / Q000235:genetics / Q000276:immunology; D011487:Protein Conformation; D015202:Protein Engineering; D014614:Vaccines, Synthetic / Q000276:immunology*; D058425:Vaccines, Virus-Like Particle
chemical_list: D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007252:Influenza Vaccines; D014614:Vaccines, Synthetic; D058425:Vaccines, Virus-Like Particle
keywords: Biomolecular engineering; Influenza; Modular; Receptor; VLP; Virus-like particle