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Expression of the Apx toxins ofActinobacillus pleuropneumoniae inSaccharomyces cerevisiae and its induction of immune response in mice
Authors:Seung-Moon Park  Eun-Jin Choi  Tae-Ho Kwon  Yong-Suk Jang  Han-Sang Yoo  Woo Bong Choi  Bong-Kyun Park  Dae-Hyuk Kim
Institution:1. Division of Biological Sciences, Basic Science Research Institute, Chonbuk National University, 561-756, Jeonju, Korea
2. Institute for Molecular Biology and Genetics, Basic Science Research Institute, Chonbuk National University, 561-756, Jeonju, Korea
3. Department of Infectious Disease, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, 151-742, Seoul, Korea
4. Department of Biotechnology and Bioengineering, Dongeui University, 614-714, Busan, Korea
5. Department of Veterinary Virology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, 151-742, Seoul, Korea
Abstract:Actinobacillus pleuropneumoniae is an important pig pathogen, which is responsible for swine pleuropneumonia, a highly contagious respiratory infection. To develop subunit vaccines forA. pleuropneumoniae infection, the Apx toxin genes,apxI andapxII, which are thought to be important for protective immunity, were expressed inSaccharomyces cerevisiae, and the induction of immune responses in mice was examined. TheapxI andapxII genes were placed under the control of a yeast hybridADH2-GPD promoter (AG), consisting of alcohol dehydrogenase II (ADH2) and theGPD promoter. Western blot analysis confirmed that both toxins were successfully expressed in the yeast. The ApxIA and ApxIIA-specific IgG antibody response assays showed dose dependent increases in the antigen-specific IgG antibody titers. The challenge test revealed that ninety percent of the mice immunized with ApxIIA or a mixture of ApxIA and ApxIIA, and sixty percent of mice immunized with ApxIA survived, while none of those in the control groups survived longer than 36 h. These results suggest that vaccination of the yeast expressing the ApxI and ApxII antigens is effective for the induction of protective immune responses againstA. pleuropneumoniae infections in mice.
Keywords:Actinobacillus pleuropneumoniae                      Saccharomyces cerevisiae            ApxIA  ApxIIA  vaccines
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