• Vaccine Name: DNA vaccine co-expressing A85A and caspase-3
• Target Pathogen: Mycobacterium tuberculosis
• Target Disease: Tuberculosis
• Vaccine Ontology ID: VO_0004148
• Type: DNA vaccine
• Antigen: The antigens for this vaccine were wild-type or mutated caspase-3 inserted into a plasmid that encodes mycolyl transferase antigen 85A (Ag85A) (Gartner et al., 2008).
• FbpA (Ag85A) gene engineering:
  • Type: DNA vaccine construction
  • Description:
  • Detailed Gene Information: Click Here.
• DNA vaccine plasmid:
  • DNA vaccine plasmid name:
  • DNA vaccine plasmid VO ID: VO_0000086
• Preparation: Native Ag85A was purified from BCG culure and recombinant his-tagged Ag85A was purified by affinity chromatography. Dual expression-vectors were created encoding mature or secreted Ag85A and either mutant or wild-type caspase-3 (Gartner et al., 2008).

Mouse Response

• Host Strain: (C57BL/6 × DBA/2)F1 (H-2b/d) or BALB/c (H-2d)
• Vaccination Protocol: Mice were injected four times intramuscularly with 100 micrograms of pDNA with at least three weeks between injections. As negative controls, mice were injected with 100 micrograms of empty pBudCE4.1 vector or left untreated. As a positive control for the challenge experiments, mice were vaccinated intravenously with M. bovis BCG strain GL2 along with the first vaccination or 5 weeks pre-vaccination (Gartner et al., 2008).
• Immune Response: Ag85A specific antibody titers were very low in mice vaccinated with the wild-type p-sAg85A–csp, but antibody titers in mice vaccinated with mutant p-sAg85A–csp were of a greater magnitude. IFN-γ and IL-2 production in mice vaccinated with plasmids co-expressing mAg85A and a caspase gene were similar to or lower than production in mice vaccinated with p-sAg85A. Vaccination with the wild-type caspase-encoding plasmids was not effective in inducing Ag85A specific CTL activity (Gartner et al., 2008).
• Challenge Protocol: Mice were challenged five weeks after their final vaccination with 2 x 10^5 CFUof M. tuberculosis H37Rv. Results were measured using a luminescence assay with a count of relative light units (RLU) (Gartner et al., 2008).
• Efficacy: The challenge revealed that mice vaccinated with mutant p-sAg85A–csp showed higher secretion of IL-6, IL-10 and IL-17A in their lungs, spontaneously and/or after antigenic stimulation.However, spontaneous IFN-γ production was highest in mice vaccinated with wild-type p-sAg85A–csp. Ag85A specific IL-2 and IFN-γ responses were low in spleen of BCG vaccinated mice, but high Th1 responses could be could be seen after stimulation with whole BCG culture filtrate in these mice (Gartner et al., 2008).
• Information about this animal model: Mouse Model for TB research