• Vaccine Name: C. muridarum DNA vaccine encoding TC0767/768
• Target Pathogen: Chlamydia muridarum
• Vaccine Ontology ID: VO_0011524
• Type: DNA vaccine
• Status: Research
• Antigen: C. muridarum TC0767/768
• TC0767 hypothetical protein gene engineering:
  • Type: DNA vaccine construction
  • Description: The C. muridarum genomic expression library was created by ligating fragments of C. muridarum DNA into the novel expression vector, pCI30. Eighteen recombinant C. muridarum clones were selected for immunizations and these were grown on LB agar. Cultures were grown for 18 h at 37 °C with shaking at 225 rpm. Following this, 1 mL of the starter culture was used to inoculate 80 mL of selective LB broth and the cultures again incubated for 18 h at 37 °C with vigorous shaking. DNA was then extracted from cell pellets using Roche Genopure Plasmid Midi kits (Roche Diagnostics Australia Pty Ltd.) (McNeilly et al., 2007).
  • Detailed Gene Information: Click Here.
• TC0768 hypothetical protein gene engineering:
  • Type: DNA vaccine construction
  • Description: The C. muridarum genomic expression library was created by ligating fragments of C. muridarum DNA into the novel expression vector, pCI30. Eighteen recombinant C. muridarum clones were selected for immunizations and these were grown on LB agar. Cultures were grown for 18 h at 37 °C with shaking at 225 rpm. Following this, 1 mL of the starter culture was used to inoculate 80 mL of selective LB broth and the cultures again incubated for 18 h at 37 °C with vigorous shaking. DNA was then extracted from cell pellets using Roche Genopure Plasmid Midi kits (Roche Diagnostics Australia Pty Ltd.) (McNeilly et al., 2007).
  • Detailed Gene Information: Click Here.
• DNA vaccine plasmid:
  • DNA vaccine plasmid name:
  • DNA vaccine plasmid VO ID: VO_0005028
• Immunization Route: intra-abdominal injection

Mouse Response

• Host Strain: BALB/c
• Vaccination Protocol: Mice were anaesthetised via inhalation of 4% isofluorane (Abbott Australasia, NSW, Australia) and the abdominal fur of each mouse was removed with electric clippers. The barrel liner of the Helios gene gun was held directly against the abdominal skin and a DNA/microcarrier shot delivered using a helium pressure of 400 psi. Each animal received two shots with the appropriate DNA/microcarrier preparation, resulting in administration of approximately 2 μg of DNA. Each mouse was immunized three times at 3-week intervals. Positive control mice received 2.5 mg of medroxyprogesterone acetate (Ralovera) (Kenral Division of Pharmacia Australia Pty Limited, Rydalmere, Australia) subcutaneously 7 days prior to receiving a dose of 5 × 10^4 live C. muridarum elementary bodies (McNeilly et al., 2007).
• Challenge Protocol: All animals were challenged intra-vaginally with 5 × 10^4 live C. muridarum elementary bodies, 10 days following the final gene gun immunization, as described for the positive control mice. To assess the level of infection, cervico-vaginal swabs were collected from each animal every 3 days following the intra-vaginal challenge for a period of 3 weeks (McNeilly et al., 2007).
• Efficacy: The groups of female BALB/c mice immunized intra-abdominally with TC0767/768 showed significant levels of protection when compared to negative control animals (McNeilly et al., 2007).