Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Multi-Immunogenic Outer Membrane Vesicles Derived from a MsbB-Deficient Salmonella enterica Serovar Typhimurium Mutant

  • Lee, Sang-Rae (The National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Sang-Hyun (The National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Jeong, Kang-Jin (The National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Keun-Su (The National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Young-Hyun (The National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Sung-Jin (The National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, E-Kyune (The National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Jung-Woo (Department of Animal Resources and Science, Dankook University) ;
  • Chang, Kyu-Tae (The National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB))
  • Published : 2009.10.31
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Abstract

To develop low endotoxic and multi-immunogenic outer membrane vesicles (OMVs), a deletion mutant of the msbB gene in Salmonella enterica serovar Typhimurium (S. Typhimurium) was used as a source of low endotoxic OMV, and an expression vector of the canine parvovirus (CPV) VP2 epitope fused to the bacterial OmpA protein was constructed and transformed into the Salmonella ${\Delta}msbB$ mutant. In a lethality test, BALB/c mice injected intraperitoneally with the Salmonella ${\Delta}msbB$ mutant survived for 7 days, whereas mice injected intraperitoneally with the wild type survived for 3 days. Moreover, all mice inoculated orally with the ${\Delta}msbB$ mutant survived for 30 days, but 80% of mice inoculated orally with the wild type survived. The OmpA::CPV VP2 epitope fusion protein was expressed successfully and associated with the outer membrane and OMV fractions from the mutant S. Typhimurium transformed with the fusion protein-expressing vector. In immunogenicity tests, sera obtained from the mice immunized with either the Salmonella msbB mutant or its OMVs containing the OmpA::CPV VP2 epitope showed bactericidal activities against wild-type S. Typhimurium and contained specific antibodies to the CPV VP2 epitope. In the hemagglutination inhibition (HI) assay as a measurement of CPV-neutralizing activity in the immune sera, there was an 8-fold increase of HI titer in the OMV-immunized group compared with the control. These results suggested that the CPV-neutralizing antibody response was raised by immunization with OMV containing the OmpA::CPV VP2 epitope, as well as the protective immune response against S. Typhimurium in BALB/c mice.

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