• Vaccine Name: Recombinant O. anthropi 49237SOD
• Target Pathogen: Brucella spp.
• Target Disease: Brucellosis
• Vaccine Ontology ID: VO_0000407
• Type: Recombinant vector vaccine
• Adjuvant:
  • Adjuvant name:
  • VO adjuvant ID: VO_0001237
• Preparation: Ochrobactrum anthropi strain 47237 was originally isolated from soil. Plasmids pBBSOD and pBBR1MCS were electroporated into electrocompetent O. anthropi strain 49237 using a protocol described previously for Brucella. Colonies of the O. anthropi strain 49237 containing pBBSOD or pBBR1MCS (designated O. anthropi strain 49237SOD or 49237pBB, respectively) were selected from TSA plates containing chloramphenicol at a concentration of 30 μg/ml (He et al., 2002)
• Virulence: O. anthropi strains are rarely pathogenic to humans (He et al., 2002)
• Description: Ochrobactrum anthropi is very closely related to Brucella, and has been used as a vaccine or vaccine vector for Brucellosis (He et al., 2002).

Mouse Response

• Vaccination Protocol: Experiment 1 (Exp. 1):
  Groups of eight mice were inoculated (i.p.) with either saline (negative control), B. abortus strain RB51 (2×10^8 CFU/mouse; positive control), recombinant B. abortus strain RB51SOD (2×108 CFU/mouse; positive control), O. anthropi 49237 (5×10^8 CFU/mouse), O. anthropi 49237pBB (5 × 108 CFU/mouse), or O. anthropi 49237SOD (5×10^8 CFU/mouse). At 2, 4, and 6 weeks after inoculation, three mice from each group were bled retroorbitally. The sera obtained were stored at −40°C until use in an enzyme-linked immunosorbent assay (ELISA) or for Western blot analysis.
  
  In a second experiment (Exp. 2) whether multiple injections were required for protection was asessed . Groups of five mice were immunized i.p. with either saline, B. abortus strain RB51, or O. anthropi 49237SOD at the doses described above. After two weeks, mice inoculated with O. anthropi 49237SOD were reimmunized with O. anthropi 49237/SOD (5×10^8 CFU/mouse). To determine whether different doses influenced the results of protection, four groups of five mice each were inoculated with O. anthropi 49237 at four different doses: 5×10^8, 5×10^7, 5×10^6, and 5×10^5 CFU/mouse. A fifth group (five mice) was inoculated i.p. with saline as negative control. Synthetic CpG-containing oligodeoxynucleotides (CpG-ODN) were administered to group four as an immunostimulatory adjuvant. One group (eight mice) was inoculated i.p. with O. anthropi strain 49237 (5×10^8 CFU/mouse); one group with strain 49237 (5×10^8 CFU/mouse) and CpG adjuvant, one group with strain 49237SOD alone (5×10^8 CFU/mouse), and one group with strain 49237SOD (5×10^8 CFU/mouse) and CpG adjuvant. The CpG adjuvant (10 nmol) was administered i.p. 4 h before inoculation and again at the time of inoculation of the bacterial strains. Three groups of mice (eight/group) served as controls and were inoculated with saline alone, CpG-ODN alone, or E. coli DH5α (10^6 CFU/mouse) and CpG-ODN. The mice were bled at 2, 4, and 6 weeks after inoculation (He et al., 2002).
• Challenge Protocol: Six weeks after inoculation, mice (5) from each group were challenged i.p. with 2×10^4 CFU of virulent B. abortus 2308/mouse. The mice were sacrificed after two weeks. Their spleens were homogenized, and spleens from the dilutions were plated to determine the numbers of Brucella CFU per spleen. The remaining unchallenged mice (each group) were sacrificed 6 to 8 weeks postinoculation, and spleen cells collected for cell culture in vitro (He et al., 2002).