Development of molecular clamp stabilized hemagglutinin vaccines for Influenza A viruses

Abstract

Influenza viruses cause a significant number of infections and deaths annually. In addition to seasonal infections, the risk of an influenza virus pandemic emerging is extremely high owing to the large reservoir of diverse influenza viruses found in animals and the co-circulation of many influenza subtypes which can reassort into novel strains. Development of a universal influenza vaccine has proven extremely challenging. In the absence of such a vaccine, rapid response technologies provide the best potential to counter a novel influenza outbreak. Here, we demonstrate that a modular trimerization domain known as the molecular clamp allows the efficient production and purification of conformationally stabilised prefusion hemagglutinin (HA) from a diverse range of influenza A subtypes. These clamp-stabilised HA proteins provided robust protection from homologous virus challenge in mouse and ferret models and some cross protection against heterologous virus challenge. This work provides a proof-of-concept for clamp-stabilised HA vaccines as a tool for rapid response vaccine development against future influenza A virus pandemics.

Protein Nanoparticle-Mediated Delivery of Recombinant Influenza Hemagglutinin Enhances Immunogenicity and Breadth of the Antibody Response

“…Our model HA antigen, H1 from A/California/7/2009 (Sino Biological), was His-tagged at the C-terminus, expressed in HEK293T cells, and lacked the transmembrane domain and cytoplasmic tail of native hemagglutinin. The protein also lacks an engineered trimerization domain, and size exclusion chromatography (SEC) comparing the elution profiles of this soluble H1 shows elution profiles consistent with previously reported monomers of H1 , and other HA subtypes (H3, H5), − rather than their trimers (Figure SI-4). Although HA trimers may present quaternary epitopes not found in monomers and thereby elicit antibodies from a broader repertoire of B cell clones, , we aim to construct H1–E2 using HA monomers to facilitate synthesis, which is an important consideration for future nanoparticle vaccine scale-up.…”

Design and Evolution of Enhanced Peptide–Peptide Ligation for Modular Transglutaminase Assembly

“…The stable association of SnoopLigase2 with SnoopTag2:DogTag2 is not surprising in light of our selection approach, which did not enforce multiple turnovers. However, in previous applications of SnoopLigase for enzyme resilience and vaccine development, SnoopLigase remaining bound did not impair functional activity of the ligated partners. , There are a range of ways to trimerize proteins, but homotrimers are much more common than heterotrimers. − In addition, coiled-coil-based heteromerization units can have impaired solubility when expressed in the absence of their cognate partners and often have unintended homodimerization. , Therefore, the SnoopLigase2 system may find application in the general challenge of assembling three separately expressed components into a precise and stable complex.…”

A single immunization with H5N1 virus-like particle vaccine protects chickens against divergent H5N1 influenza viruses and vaccine efficacy is determined by adjuvant and dosage

“…These immunological components can be incorporated into H5N1 VLP to enhance the cross-protection capacity of H5N1 VLP vaccines. Strategies to enhance the immunogenicity of HA can also be chosen to improve the immune responses of influenza VLP vaccines [ 43 , 44 ]. Strategies of construction of the multivalent VLP, mucosal immunization, and combined nanoparticle were demonstrated to enhance the cross-protective efficacy of influenza vaccines [ 23 , 45 , 46 ].…”