• Vaccine Ontology ID: VO_0011533
• Type: DNA vaccine
• Status: Research
• Antigen: C. muridarum TC0439 adherence factor
• TC0439 adherence factor 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.
• Vector: pCI30
• Immunization Route: Subcutaneous

• Vaccine Ontology ID: VO_0011534
• Type: DNA vaccine
• Status: Research
• Antigen: C. muridarum TC0512 outer membrane protein, putative
• TC0512 outer membrane protein, putative 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.
• Vector: pCI30 (McNeilly et al., 2007)
• Immunization Route: Subcutaneous injection

• Vaccine Ontology ID: VO_0011522
• Type: DNA vaccine
• Status: Research
• Antigen: C. muridarum TC0757
• TC0757 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.
• Vector: pCI30
• Immunization Route: Subcutaneous injection

• 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.
• Vector: pCI30
• Immunization Route: intra-abdominal injection

• Vaccine Ontology ID: VO_0011453
• Type: Subunit vaccine
• Status: Research
• Antigen: C. muridarum major outer membrane protein MOMP
• MOMP gene engineering:
  • Type: Recombinant protein preparation
  • Description: Recombinant maltose binding protein-MOMP (MBP-MOMP) was cloned into the bacterial expression vector pMAL-c2, and used to transform Escherichia coli (DH5α[pMMM3]). The MBP-MOMP was purified by passage through a PD-10 column (Amersham Biosciences, NSW, Australia), followed by a Sephadex G-75 column (Sigma–Aldrich) (Skelding et al., 2006).
  • Detailed Gene Information: Click Here.
• Adjuvant:
  • VO ID: VO_0000143
• Adjuvant:
  • VO ID: VO_0001237
• Immunization Route: Intranasal immunization

• Vaccine Ontology ID: VO_0011456
• Type: Subunit vaccine
• Status: Research
• Antigen: C. muridarum polymorphic membrane protein E/F family protein
• PmpE/F-2 gene engineering:
  • Type: Recombinant protein preparation
  • Description: The source proteins containing the MHC II binding Chlamydia peptides were cloned, expressed and purified as follows: pmpE/F-2 and gap DNA fragments were generated by PCR using genomic DNA isolated from C. muridarum. The PCR products were purified and cloned into pET32a (Novagen) for pmpE/F-2 and gap after restriction enzyme digestion with BamHI/NotI using standard molecular biology techniques. Only the first half of the gene (25–575) was cloned into the vector for expression (Yu et al., 2009).
  • Detailed Gene Information: Click Here.
• Adjuvant:
  • VO ID: VO_0000142
  • Description: Incomplete Freunds Adjuvant (Sigma)
• Immunization Route: Intravenous injection (i.v.)

• Vaccine Ontology ID: VO_0011458
• Type: Subunit vaccine
• Status: Research
• Antigen: C. muridarum polymorphic membrane protein G family protein
• PmpG-1 gene engineering:
  • Type: Recombinant protein preparation
  • Description: The source proteins containing the MHC II binding Chlamydia peptides were cloned, expressed and purified as follows: pmpG-1 and gap DNA fragments were generated by PCR using genomic DNA isolated from C. muridarum. The PCR products were purified and cloned into pET32a (Novagen) for pmpG-1 and gap after restriction enzyme digestion with BamHI/NotI using standard molecular biology techniques. Only the first half of the gene (25–500) was cloned into the vector for expression (Yu et al., 2009).
  • Detailed Gene Information: Click Here.
• Adjuvant:
  • VO ID: VO_0000142
  • Description: Incomplete Freunds Adjuvant (Sigma)
• Immunization Route: Intravenous injection (i.v.)

• Vaccine Ontology ID: VO_0004257
• Type: Subunit vaccine
• Status: Research
• Antigen: Chlamydia proteins PmpG-1, PmpE/F-2, and MOMP (Yu et al., 2010).
• Adjuvant:
  • VO ID: VO_0001334
  • Description: Abisco-100
• Immunization Route: subcutaneous injection

• Vaccine Ontology ID: VO_0011437
• Type: Subunit vaccine
• Status: Research
• Antigen: C. muridarum rplF
• RplF gene engineering:
  • Type: Recombinant protein preparation
  • Description: The source proteins containing the MHC II binding Chlamydia peptides were cloned, expressed and purified as follows: rplF DNA fragments were generated by PCR using genomic DNA isolated from C. muridarum. The PCR products were purified and cloned into pGEX-6P-3 (GE Healthcare) after restriction enzyme digestion with BamHI/NotI using standard molecular biology techniques (Yu et al., 2009).
  • Detailed Gene Information: Click Here.
• Adjuvant:
  • VO ID: VO_0000142
  • Description: Incomplete Freunds Adjuvant (Sigma)
• Immunization Route: Intravenous injection (i.v.)

• Vaccine Ontology ID: VO_0004202
• Type: Subunit vaccine
• Status: Research
• Antigen: purified C. muridarum Tarp
• Tarp gene engineering:
  • Type: Recombinant protein preparation
  • Detailed Gene Information: Click Here.
• Adjuvant:
  • VO ID: VO_0001237
• Adjuvant:
  • VO ID: VO_0000142
• Immunization Route: Intramuscular injection (i.m.)

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 TC0439 showed significant levels of protection when compared to negative control animals (McNeilly et al., 2007).

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 TC0512 showed significant levels of protection when compared to negative control animals. At 9 days following challenge TC0512 showed a 73% reduction in the number of recoverable Chlamydia compared with vector only immunized controls (McNeilly et al., 2007).

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 TC0757 showed significant levels of protection when compared to negative control animals (McNeilly et al., 2007).

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).

Mouse Response

• Host Strain: BALB/c
• Vaccination Protocol: Groups of five mice were immunized on days 0, 7, 14, and 35 via either the IN or TCI route. One group of five mice were used as a control, receiving no immunization solution. Mice immunized intranasally were anaesthetised via inhalation of 4% isofluorane (Abbott Australasia, NSW, Australia), and 10 μl of immunization solution containing 100 μg MOMP, admixed with 10 μg CpG-ODN 1826 (5′-TCC ATG ACG TTC CTG ACG TT-3′) (Geneworks, SA, Australia), and 10 μg cholera toxin (Sigma–Aldrich) was gently pipetted onto each nare (5 μl per nare) (Skelding et al., 2006).
• Challenge Protocol: Five days after the final boost, mice were infected intranasally with C. muridarum. Mice were anaesthetised with 4% isoflurane and 10^3 IFUs were gently pipetted onto the nares. Mice were weighed daily for 10 days then sacrificed 10 days post-infection via a sodium pentobarbital overdose (Nembutal, Abbott Australasia). The lungs from the mice were either perfused and fixed for histopathology or collected for determination of C. muridarum levels (Skelding et al., 2006).
• Efficacy: Both intranasal and transcutaneous immunization protected mice against respiratory challenge with Chlamydia (Skelding et al., 2006).
• Host Ifng (Interferon gamma) response
  • Description: Both TCI and IN immunization resulted in greatly increased IFN-γ gene expression in stimulated lymph node cells (stimulated with MOMP) as compared to non-immunized mice (Skelding et al., 2006).
  • Detailed Gene Information: Click Here.
• Host IgA response
  • Description: Both routes of immunization induced a significant MOMP-specific IgA response in serum with the highest IgA response seen in animals immunized intranasally. These results were compared 7 days post-immunization to non-immunized control mice (Skelding et al., 2006).
  • Detailed Gene Information: Click Here.
• Host IgG response
  • Description: Both routes of immunization (TCI and IN) resulted in levels of MOMP-specific IgG in serum that were significantly higher than that in serum of non-immunized control mice. There was no significant difference between serum IgG levels in animals immunized by either route. These results were gathered 7 days post-immunization (Skelding et al., 2006).
  • Detailed Gene Information: Click Here.

Mouse Response

• Host Strain: C57BL/6
• Vaccination Protocol: Mice were vaccinated three times with a 2-week interval, intravenously (i.v.) into the tail veins with 1 × 10^6 DCs transfected with Chlamydia protein PmpE/F-225–575 in 200 μl of PBS. DCs pulsed with live EB or GST protein was used as positive or negative controls respectively (Yu et al., 2009).
• Challenge Protocol: Two weeks after the final immunization, five to ten mice from each group were intranasally (i.n.) challenged with 2000 IFU of C. muridarum. Weight loss was monitored each or every two days. On 10 day after i.n. challenge, the mice were euthanized and the lungs were collected for Chlamydia titration (Yu et al., 2009).
• Efficacy: PmpE/F-2(25-575) immunized mice exhibited significant resistance to challenge infection. In addition, pmpE/F-2 was able to engender protective immunity against challenge with C. muridarum (Yu et al., 2009).

Mouse Response

• Host Strain: C57BL/6
• Vaccination Protocol: Mice were vaccinated three times with a 2-week interval, intravenously (i.v.) into the tail veins with 1 × 10^6 DCs transfected with Chlamydia protein PmpG-1 25–500 in 200 μl of PBS. DCs pulsed with live EB or GST protein was used as positive or negative controls respectively (Yu et al., 2009).
• Challenge Protocol: Two weeks after the final immunization, five to ten mice from each group were intranasally (i.n.) challenged with 2000 IFU of C. muridarum. Weight loss was monitored each or every two days. On 10 day after i.n. challenge, the mice were euthanized and the lungs were collected for Chlamydia titration (Yu et al., 2009).
• Efficacy: Overall, vaccination with DCs transfected with PmpG-1(25-500) exhibited the greatest degree of protective immunity among the four Chlamydia Ags tested (Yu et al., 2009).

Mouse Response

• Host Strain: C57BL/6 and BALB/c
• Vaccination Protocol: Mice were vaccinated 3 times subcutaneously with PmpG-1 plus PmpE/F-2 plus MOMP plus AbISCO-100 (G+F+M+AbISCO) (Yu et al., 2010).
• Challenge Protocol: Two weeks after the final immunization, the mice in the three animal trials described above were then challenged with live EBs for protection evaluation (Yu et al., 2010).
• Efficacy: The combination of three Chlamydia proteins formulated with AbISCO exhibited significant protection at day 13 compared to the group with adjuvant alone (Yu et al., 2010).

Mouse Response

• Host Strain: C57BL/6
• Vaccination Protocol: Mice were vaccinated three times with a 2-week interval, intravenously (i.v.) into the tail veins with 1 × 10^6 DCs transfected with Chlamydia protein RplF in 200 μl of PBS. DCs pulsed with live EB or GST protein was used as positive or negative controls respectively (Yu et al., 2009).
• Challenge Protocol: Two weeks after the final immunization, five to ten mice from each group were intranasally (i.n.) challenged with 2000 IFU of C. muridarum. Weight loss was monitored each or every two days. On 10 day after i.n. challenge, the mice were euthanized and the lungs were collected for Chlamydia titration (Yu et al., 2009).
• Efficacy: RplF was able to engender protective immunity against challenge with C. muridarum (Yu et al., 2009).

Mouse Response

• Host Strain: BALB/c
• Vaccination Protocol: Mice were immunized intramuscularly with purified C. muridarum Tarp emulsified in CPG-IFA adjuvant or just adjuvant (as a control). Mice were immunized in the hind legs 3 times with an interval of 3 weeks between the first and second injections and 9 days between the second and third injections (Wang et al., 2009).
• Challenge Protocol: Thirty days after the third immunization, each mouse was inoculated intravaginally with 1 × 10^4 IFUs of live C. muridarum organisms in 20μl of SPG (sucrose-phosphate-glutamate buffer). Five days prior to infection, each mouse was injected with 2.5mg Depo-provera subcutaneously to synchronize menstrual cycle and increase mouse susceptibility to chlamydial infection (Wang et al., 2009).
• Efficacy: Immunization of mice with Tarp induced Th1-dominant immunity that significantly reduced the shedding of live organisms from the lower genital tract and attenuated inflammatory pathologies in the fallopian tube tissues. Tarp, an immunodominant antigen identified by human antisera, can induce protective immunity against chlamydial infection and pathology in mice (Wang et al., 2009).