• Vaccine Name: Influenza virus CTA1-3M2e-DD protein vaccine
• Target Pathogen: Influenza virus
• Target Disease: Influenza (flu)
• Vaccine Ontology ID: VO_0002980
• Type: Subunit vaccine
• Status: Research
• Antigen: Influenza A virus (A/Puerto Rico/8/34(H1N1)) matrix protein 2 (M2)
• M2 from Influenza A virus (A/Puerto Rico/8/34(H1N1)) gene engineering:
  • Type: Recombinant protein preparation
  • Description: The fusion proteins were expressed in E. coli DH5 cells, transformed with the expression vectors for the CTA1-DD, CTA1-M2e-DD or the CTA1-3M2e-DD fusion proteins, and grown in 500 ml cultures over night in SYPPG medium with 100 μg/ml carbenicillin, at 37 °C. The cells were harvested by centrifugation and the fusion proteins, produced as inclusion bodies, were washed before extraction by treatment with 8 M urea. After refolding the proteins by slowly diluting them 35–40x in Tris–HCl pH 7.4 at +4 °C, the fusion proteins were purified in two steps, by ion exchange and size exclusion chromatography. After concentration and sterile filtration the purified fusion proteins were stored in PBS at −80 °C until use (Eliasson et al., 2008).
  • Detailed Gene Information: Click Here.
• Adjuvant:
  • Adjuvant name:
  • VO adjuvant ID: VO_0001290
  • Description: CTA1-DD
• Immunization Route: Intranasal

Mouse Response

• Host Strain: BALB/c
• Vaccination Protocol: Age- and sex matched mice were immunized intranasally (i.n.) twice, 3 weeks apart, with 20–50 μl containing 10 μg of the M2e-HBc particle 1818 admixed with 5 μg CTA1-DD-adjuvant, 10 μg of HBc admixed with 5 μg CTA1-DD-adjuvant, a range of doses as indicated were tested; CTA1-M2e-DD, CTA1-3M2e-DD (carrying three copies of M2e), CTA1-DD adjuvant alone or PBS. Groups with 3–12 individuals in each experiment were used as indicated (Eliasson et al., 2008).
• Challenge Protocol: Challenge experiments were performed in Ghent. Before the first, and 2 weeks after each immunization, blood samples were collected from the ventral tail vein. The final bleeding of surviving mice was performed 2 weeks after challenge (Eliasson et al., 2008).
• Efficacy: A targeted fusion protein based on the CTA1-DD adjuvant and containing tandem repeats of the matrix protein 2 (M2e) ectodomain epitope derived from Influenza A virus (A/Puerto Rico/8/34(H1N1)), CTA1-3M2e-DD, confers strong protective immunity against a potentially lethal challenge infection with influenza virus in mice. The formulation was highly effective for mucosal immunizations and promoted high M2e-specific serum IgG and mucosal IgA antibody titers and an hitherto unknown anti-M2e CD4 T cell immunity (Eliasson et al., 2008).
• Host IgA response
  • Description: The CTA1-3M2e-DD construct stimulated significant anti-M2e IgA antibody titers in bronchial lavage. By contrast, soluble M2e-peptide, given even at high doses, 100 μg, together with CTA1-DD (WT), failed completely to stimulate specific serum antibody titers. Antibody titer experiments were repeated at least twice with similar results (Eliasson et al., 2008).
  • Detailed Gene Information: Click Here.
• Host Ighg1 response
  • Description: CTA1-M2e-DD elicited a strong IgG1 response. By contrast, soluble M2e-peptide, given even at high doses, 100 μg, together with CTA1-DD (WT), failed completely to stimulate specific serum antibody titers. Antibody titer experiments were repeated at least twice with similar results (Eliasson et al., 2008).
  • Detailed Gene Information: Click Here.
• Host Ighv1-9 response
  • Description: CTA1-M2e-DD elicited a strong IgG2a response. By contrast, soluble M2e-peptide, given even at high doses, 100 μg, together with CTA1-DD (WT), failed completely to stimulate specific serum antibody titers. Antibody titer experiments were repeated at least twice with similar results (Eliasson et al., 2008).
  • Detailed Gene Information: Click Here.