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Vaccine Detail

NPAP Brucella vaccine
Vaccine Information
  • Vaccine Name: NPAP Brucella vaccine
  • Target Pathogen: Brucella spp.
  • Target Disease: Brucellosis
  • Product Name: Brucella vaccine using nonpathogenic alphaproteobacteria
  • Vaccine Ontology ID: VO_0000450
  • Type: Inactivated or "killed" vaccine
  • Antigen: The angtiens for this vaccine were Ochrobactrum anthropi, Sinorhizobium meliloti, Mesorhizobium loti, Agrobacterium tumefaciens, or Brucella melitensis H38. These were used because provious findings indicate that Brucella antigens and those from Nonpathogenic Alphaproteobacteria (NPAP) are cross-recognized by the immune system (Delpino et al., 2007a).
  • Adjuvant:
  • Preparation: Bacterial cultures were grown on plates, then centrifuged at 15,000 x g. Bacterial pellets were washed with PBS, then the bacteria were heat-killed. These suspensions were centrifuged at 15,000 x g again, then washed with Tris buffer. The cells were suspended in Tris buffer , then disrupted with a French press. Bacterial cells were broken by two passages and then digested for 1 hour with D. Nase and R. Nase. Unbroken cells were seperated by centrifugation. Particulate matter was pelleted by centrifugation, and the resulting supernatent was stored for vaccine (Delpino et al., 2007a).
  • Virulence: Not noted.
Host Response

Mouse Response

  • Host Strain: BALB/c
  • Vaccination Protocol: Mice were immunized with 3 doses (at 2 week intervals) of heat-killed bacteria (1 x 10^9 CFU per dose) mixed with adjuvant and administered subcutaneously in the back or with 3 doses of CYTs administered intraperitoneally. Control groups received PBS. The Brucella group was immunized with B. Melitensis H38 (Delpino et al., 2007a).
  • Persistence: Not noted.
  • Immune Response: Subcutaneous immunization with with heat-killed bacteria elicited a strong antobody response against CYT antigens. Antibody levels were highest after the second immunization, in most cases. In mice immunized with nonpathogenic species. Anti-Brucella antibody levels were augmented in all groups compared to preimmunization levels (Delpino et al., 2007a).
  • Challenge Protocol: One month after the final immunization, mice were challenged intravenously with 1.3 x 10^4 CFU of live B. abortus 2308. The mice were killed 30 days later and had their spleens analyzed and plated (Delpino et al., 2007)
  • Efficacy: Cross-reacting anibodies were detected in serum samples obtained at the moment of challenge in all groups of mice. Spleen counts were significantly lower in immunized mice than in mice injected with PBS. Protection levels for nonpathogenic bacteria were lower than that obtained with Brucella immunization . Counts of CFU in mice immunized with B. melitensis were significantly lower than those of mice immunized with any other bacteria. Anti-brucella antibodies were detected in all groups 30 days postchallenge at levels similar to those found at the moment of challenge (Delpino et al., 2007a).

Mouse Response

  • Host Strain: BALB/c
  • Vaccination Protocol: Two separate experiments of oral immunization were performed. Mice were inoculated intragastrically at weekly intervals with three doses of live NPAP(1 x 10^8 CFU) or heat-killed B. abortus (HKBA) strain 2308 (1 x 10^8 CFU)or PBS by an intragastric feeding tube (Delpino et al., 2007a).
  • Persistence: Not noted.
  • Immune Response: Anti-Brucella IgG levels were significatly higher than preinfection levels in all groups of mice infected with NPAP. Mice infected with HKBA exhibited a significant rise in anti-Brucella IgG in serum. No significant increase in IgA was detected at the time of challenge for mice immunized with HKBA or infected with NPAP (Delpino et al., 2007a).
  • Challenge Protocol: Twenty days after the last infecting dose, mice were challenged by the same route with live B. abortus 2308 (Delpino et al., 2007a).
  • Efficacy: Serum levels of anti-Brucella IgA increased significantly in mice previously infected with O. anthropi and in mice orally infected with HKBA. Fecal levels of anti-Brucella IgA also increased after challenge (Delpino et al., 2007a).
References
Delpino et al., 2007a: Delpino MV, Estein SM, Fossati CA, Baldi PC. Partial protection against Brucella infection in mice by immunization with nonpathogenic alphaproteobacteria. Clinical and vaccine immunology : CVI. 2007; 14(10); 1296-1301. [PubMed: 17715332].