Development of a candidate influenza vaccine based on virus-like particles displaying influenza M2e peptide into the immunodominant loop region of hepatitis B core antigen: Insertion of multiple copies of M2e increases immunogenicity and protective efficiency.
The extracellular domain of the transmembrane protein M2 (M2e) of influenza A virus is a promising target for the development of "universal" vaccines against influenza. M2e is a poor immunogen by itself; however, when M2e is linked to an appropriate carrier, such as hepatitis B virus core (HBc) particles, it becomes highly immunogenic. Insertions of target peptides into the surface-exposed major immunodominant loop region (MIR) of the HBc antigen are especially immunogenic, but such insertions often affect the protein folding and formation of recombinant virus-like particles. To facilitate an appropriate conformation of the M2e insert, we introduced flexible linkers at the junction points between the insert and flanking HBc sequences. This approach allowed the construction of recombinant HBc particles carrying 1, 2 and 4 copies of M2e in the MIR region. These particles were produced in Escherichia coli and purified to homogeneity. The immune response and protective activity of hybrid HBc particles in mice correlated with the number of inserted M2e peptides: the highest immunogenicity and complete protection of mice against the lethal challenge by influenza virus was observed with particles carrying four copies of M2e. The possibility of the simultaneous presentation of M2e peptides from several important influenza strains on a single HBc particle could also facilitate the development of a broad-specificity vaccine efficient not only against influenza A strains of human origin but also for newly emerging strains of animal origin, such as the avian influenza.
Centre 'Bioengineering', Russian Academy of Sciences, 117312 Prosp. 60-letya Oktyabrya 7-1, Moscow, Russia. Electronic address: nravin@biengi.ac.ru.
Nikolai V
NV
Ravin
Centre 'Bioengineering', Russian Academy of Sciences, 117312 Prosp. 60-letya Oktyabrya 7-1, Moscow, Russia.
Elena A
EA
Blokhina
Centre 'Bioengineering', Russian Academy of Sciences, 117312 Prosp. 60-letya Oktyabrya 7-1, Moscow, Russia.
Victor V
VV
Kuprianov
Research Institute of Influenza, Russian Federation Ministry of Health, St. Petersburg, Russia.
Liudmila A
LA
Stepanova
Research Institute of Influenza, Russian Federation Ministry of Health, St. Petersburg, Russia.
Aram A
AA
Shaldjan
Research Institute of Influenza, Russian Federation Ministry of Health, St. Petersburg, Russia.
Anna A
AA
Kovaleva
Research Institute of Influenza, Russian Federation Ministry of Health, St. Petersburg, Russia.
Liudmila M
LM
Tsybalova
Centre 'Bioengineering', Russian Academy of Sciences, 117312 Prosp. 60-letya Oktyabrya 7-1, Moscow, Russia.
Konstantin G
KG
Skryabin
D000818:Animals; D004195:Disease Models, Animal; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015870:Gene Expression; D006512:Hepatitis B Core Antigens / Q000235:genetics; D007252:Influenza Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D008807:Mice, Inbred BALB C; D016254:Mutagenesis, Insertional; D009976:Orthomyxoviridae Infections / Q000517:prevention & control; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D016019:Survival Analysis; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D058425:Vaccines, Virus-Like Particle / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology*; D000086082:Viroporin Proteins
D000939:Epitopes; D006512:Hepatitis B Core Antigens; D007252:Influenza Vaccines; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D058425:Vaccines, Virus-Like Particle; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins; C509609:M2e-HBc influenza vaccine
HBc particle; Influenza virus; M2e; Vaccine