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Updated: Jun 11, 2025
Expression and Purification of Virus-like Particles for Vaccination
Published on: June 2, 2016
Influenza A Vaccine Candidates Based on Virus-like Particles Formed by Coat Proteins of Single-Stranded RNA Phages
Egor A Vasyagin1, Anna A Zykova1, Eugenia S Mardanova1
1Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, Russia.
Abstract:
Efficient control of influenza A infection can potentially be achieved through the development of broad-spectrum recombinant vaccines based on conserved antigens. The extracellular domain of the transmembrane protein M2 of influenza A virus (M2e) is highly conserved but poorly immunogenic and needs to be fused to an adjuvant protein or carrier virus-like particles (VLPs) to increase immunogenicity and provide protection against infection. In this study, we obtained VLPs based on capsid proteins (CPs) of single-stranded RNA phages Beihai32 and PQ465 bearing the M2e peptides. Four copies of the M2e peptide were linked to the C-terminus of the CP of phage Beihai32 and to the N and C termini of the CP of phage PQ465. The hybrid proteins, being expressed in Escherichia coli, formed spherical VLPs of about 30 nm in size. Immunogold transmission electron microscopy showed that VLPs formed by the phage PQ465 CP with a C-terminal M2e fusion present the M2e peptide on the surface. Subcutaneous immunization of mice with VLPs formed by both CPs containing four copies of the M2e peptide at the C termini induced high levels of M2e-specific IgG antibodies in serum and provided mice with protection against lethal influenza A virus challenge. In the case of an N-terminal fusion of M2e with the phage PQ465 CP, the immune response against M2e was significantly lower. CPs of phages Beihai32 and PQ465, containing four copies of the M2e peptide at their C termini, can be used to develop recombinant influenza A vaccine.
Insights
Developing a universal influenza A vaccine is crucial. This study shows that virus-like particles (VLPs) carrying the M2e peptide, derived from conserved influenza antigens, effectively protect mice against lethal influenza A virus challenge.
Area of Science:
- Virology and Immunology
- Vaccine Development
- Recombinant Protein Engineering
Background:
- Influenza A virus poses a significant public health threat, necessitating broad-spectrum vaccines.
- The M2 extracellular domain (M2e) is a conserved influenza A antigen but requires enhancement for immunogenicity.
- Virus-like particles (VLPs) serve as effective carriers for vaccine antigens to improve immune responses.
Purpose of the Study:
- To engineer and evaluate virus-like particles (VLPs) based on phage capsid proteins (CPs) displaying the M2e peptide.
- To assess the immunogenicity and protective efficacy of M2e-displaying VLPs against influenza A infection in a mouse model.
Main Methods:
- Recombinant M2e peptides were fused to capsid proteins (CPs) of single-stranded RNA phages Beihai32 and PQ465.
- Hybrid proteins were expressed in *Escherichia coli*, forming spherical VLPs approximately 30 nm in size.
- Immunogold transmission electron microscopy confirmed M2e peptide display on VLP surfaces.
- Mice were immunized subcutaneously with engineered VLPs, and immune responses and protection against lethal influenza A challenge were assessed.
Main Results:
- VLPs formed by both phage CPs with C-terminal M2e fusions induced high levels of M2e-specific IgG antibodies.
- Subcutaneous immunization with these VLPs conferred significant protection against a lethal influenza A virus challenge.
- N-terminal M2e fusion with phage PQ465 CP resulted in a significantly lower immune response against M2e.
Conclusions:
- Capsid proteins of phages Beihai32 and PQ465, when engineered with C-terminal M2e fusions, are suitable for developing recombinant influenza A vaccines.
- These M2e-displaying VLPs demonstrate potential as a basis for a broad-spectrum influenza A vaccine candidate.
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