• Vaccine Name: F. tularensis GroEL protein vaccine
• Target Pathogen: Francisella tularensis
• Target Disease: Tularemia
• Vaccine Ontology ID: VO_0011446
• Type: Subunit vaccine
• Status: Research
• Antigen: F. tularensis chaperonin GroEL (Hsp60)
• groEL gene engineering:
  • Type: Recombinant protein preparation
  • Description: Hsp60 was identified by Western blotting with a monoclonal antibody to GroEL (Sigma-Aldrich, Poole, United Kingdom). Hsp60 was excised from large-format unstained gels and electroeluted into 4× Laemmli buffer (0.1 M Tris-HCl [pH 7.3], 0.768 M glycine, 0.4% [wt/vol] SDS) by using the Hoefer gel eluter (Amersham Pharmacia, Buckinghamshire, United Kingdom) at 70 V for 2 h, following the manufacturer's instructions. The eluted product was pooled and purified by dilution (more than 20×) in ammonium bicarbonate (3.95 g/liter) and SDS (1 g/liter) and centrifuged over a dialysis membrane (10,000-molecular-weight cutoff; Vivascience, Lincoln, United Kingdom). The protein was then diluted (20×) in sterile water and centrifuged further, until significant concentration was achieved (Hartley et al., 2004).
  • Detailed Gene Information: Click Here.
• Adjuvant:
  • Adjuvant name:
  • VO adjuvant ID: VO_0001147
• Immunization Route: Intraperitoneal injection (i.p.)

Mouse Response

• Host Strain: BALB/c
• Vaccination Protocol: Groups of BALB/c mice 6 to 8 weeks old (Charles River International, Margate, United Kingdom) were immunized i.p. on days 0 and 14 with 25 μg of Hsp60/100 μl of saline with or without 0.5 μg of murine IL-12. Control groups were immunized with either IL-12 only or 100 μl of saline containing eluted protein-free gel. IL-12 was kindly provided by Roche Pharmaceuticals. Blood samples were obtained from each mouse by tail vein bleeding immediately prior to challenge (day 28). The serum was separated by microcentrifugation and stored at −20°C until analysis. Groups of mice were either challenged or culled to provide spleen cells a further 14 days later (Hartley et al., 2004).
• Challenge Protocol: F. tularensis LVS, F. tularensis subsp. holarctica HN63, and F. tularensis subsp. tularensis Schu S4 were used for challenges. The strains were harvested into PBS after growth on BCGA for 24 h at 37°C and diluted to a given optical density. Confirmation of the dose was achieved by plating out a serial dilution. Groups of mice were challenged on day 28 with either 103 or 104 MLD of LVS i.p. or 100 MLD of HN63 or 10 MLD of Schu4 subcutaneously as separate challenges. Mice were monitored for 14 days, and survival to a humane end point was recorded (Hartley et al., 2004).
• Efficacy: Researchers have shown that mice that had been immunized with purified heat shock protein 60 (Hsp60, groEL) isolated from Francisella tularensis were protected against a subsequent challenge with some strains of the bacterium (Hartley et al., 2004).