Research on pathogenic gienes and development of vaccine of Vibrio valunificus, a novel causative agent of foodborne infection

2012 Fiscal Year Final Research Report

• Project/Area Number: 22590400
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bacteriology (including Mycology)
• Research Institution: Okayama Prefectural University
• Principal Investigator: YAMAMOTO Kouichirou 岡山県立大学, 保健福祉学部, 教授 (30158274)
• Co-Investigator(Kenkyū-buntansha): NAKATA Kazue 岡山県立大学, 保健福祉学部, 助教 (60411740)
• Project Period (FY): 2010 – 2012
• Keywords: ビブリオ / バルニフィカス菌 / 感染 / 毒素 / トランスポゾン / ワクチン

Research Abstract

Vibrio vulnificus is an important pathogens of foodborn diseases and wound infection. This pathogen causes fatal septicemia rapidly in immunocompromised hosts after they contact with contaminated seawater or seafood with this organism. The mortality rate is more than 50%. For the rapid progress and the high mortality,vaccination is considered an effective way to control infection with V. vulnificus in a high risk population. We applied Signature-Tagged Mutagenesis (STM) to generate attenuated mutants from a clinically-isolated virulent V. vulnificus strain, OPU1 which was isolated from a patient. Twelve insertion mutants were selected and the virulence of these mutants were determined by the minimal lethal dose (MLD) to mice. Several mutant strains showed lower virulence compare to the parent virulent strain. The DNA sequences flanking to transposon insertion sites in these mutants were determined and searched for homologies at DNA Date Bank of Japan. The transposons inserted genes were those for IMP dehydrogenase, UDP-N-acetylgulcosamin-2-epimerase, aspartokinase, and a conserved hypothetical protein. This suggests that these genes may be involved in V. vulnificusvirulence. One of the mutants was administrated for experimental vaccination to mice. The immunized mice were challenged with the virulent parent strain. All unimmunized mice were killed within 36 hr, but almost all immunized mice survived. In this experiment, we found that the attenuated have a potentiality as live vaccine candidate.

Research Products

• [Journal Article] Bovine serum albumin significantly improves the DPPH free radicals scavenging potential of dietarypolyphenols and gallic acids (2012)
  • Author(s): Cao, H., Chen, X., and Yamamoto, K
  • Journal Title: Anti-Cancer Agents in Medicinal Chemistry Volume: 12 Pages: 940-948
• [Journal Article] Interaction of dietary polyphenols with bovine milk proteins: Molecular structure-affinity relationship and influencing bioactivity aspects (2011)
  • Author(s): Xiao, J., Mao, F., Yang, F., Zhao, F., Zhang, C., Yamamoto, K
  • Journal Title: Mol. Nutr. Food Res. Volume: 55 Pages: 1-9
• [Journal Article] Molecular structure-affinity relationship of natural polyphenols for bovine γ-globulin (2011)
  • Author(s): Xiao, J. Kai, G.,Yang, F., Liu, C.,Xu, X., Yamamoto, K
  • Journal Title: Mol. Nutr. Food Res. Volume: 55 Pages: 86-92
• [Journal Article] Structure-affinity relationship of flavones on binding to serum albumins: Effect of hydroxyl groups on ring A (2010)
  • Author(s): Xiao, J., Cao, H., Wang, Y., Yamamoto, K.,Wei, X
  • Journal Title: Mol. Nutr. Food Res. Volume: 54 Pages: 253-260
• [Presentation] Identification of genes required for Vibrio vulnificusvirulence by signature-tagged transposon mutagenesis (2013)
  • Author(s): Yamamoto, M. et al.
  • Organizer: 第86回日本細菌学会総会
  • Place of Presentation: 千葉
  • Year and Date: 20130300
• [Presentation] Response of intestinal macrophage to non-pathogenic and pathogenic bacteria.非病原性/病原性細菌に対する腸管マクロファージの応答 (2013)
  • Author(s): Nakata K., Yasuda, M., Yamamoto, M., Yamamoto, K.
  • Organizer: 第86回日本細菌学会総会
  • Place of Presentation: 千葉
  • Year and Date: 20130300
• [Presentation] STM 法による Vibrio vulnificus 病原性に必要な遺伝子のスクリーニング (2013)
  • Author(s): 山本真衣、横田憲治、小熊恵二、山本耕一郎
  • Organizer: 第86回日本細菌学会総会
  • Place of Presentation: 千葉
  • Year and Date: 20130300
• [Presentation] Transposon mutation pathogenic genes of Vibrio vulnificus (2012)
  • Author(s): Yamamoto, M., Tong, P. , Xiao, J. , Nakata, K., Yokota, K., Oguma, K., Yamamoto, K.
  • Organizer: The 47th US-Japan Cholera Panel Conference
  • Place of Presentation: Chiba, Japan
  • Year and Date: 20121200
• [Presentation] 病原性/非病原性細菌に対する腸管マクロファージの応答性 (2012)
  • Author(s): 中田和江、安田美穂、山本耕一郎、稲川裕之、杣源一郎
  • Organizer: バイオ治療法研究会
  • Year and Date: 20120000
• [Presentation] Isolation of virulence-attenuated mutants of Vibrio vulnificus by transposon-mutagenesis. In Proceedings of Bioactive Okayama 2012 (2012)
  • Author(s): Yamamoto, M., Xiao, J., Nakata, K., Yokota, K., Oguma, K., Yamamoto, K.
  • Organizer: An International Sympodium on Biologically-Active Substances.
  • Place of Presentation: Okayma, Japan
  • Year and Date: 2012-09-14
• [Presentation] Response of intestinal macrophage to non-pathogenic and pathogenic bacteria. In Proceedings of Bioactive Okayama 2012 (2012)
  • Author(s): Nakata, K., Yasuda, M., Yamamoto, K
  • Organizer: An International Symposium on Biologically-Active Substances
  • Place of Presentation: Okayma, Japan
  • Year and Date: 2012-09-14
• [Presentation] Transposon mutation of pathogenic genes of Vibrio vulnificus (2012)
  • Author(s): Yamamoto, K., Yamamoto, M., Tong, P., Xiao, J., Nakata, K., Yokota, K., Oguma, K.
  • Organizer: 日米コレラ部会(日米医学協力研究会コレラ・細菌性腸管感染症専門部会)日本側総会
  • Place of Presentation: 京都
  • Year and Date: 2012-08-06
• [Presentation] Vibrio vulnficus 溶血性に関する遺伝子 (2012)
  • Author(s): 守上加奈子,金島 健,宮本 菜津美,中田和江,山本 耕一郎.
  • Organizer: 第85回日本細菌学会総会.
  • Place of Presentation: 長崎
  • Year and Date: 2012-03-29
• [Presentation] トランスポゾン挿入変異株を用いた Vibrio vulnificus 溶血性遺伝子の研究 (2011)
  • Author(s): 守上加奈子,金島 健,宮本 菜津美,山本 耕一郎.
  • Organizer: 第64回日本細菌学会中国四国支部総会
  • Place of Presentation: 岡山
  • Year and Date: 2011-10-22
• [Presentation] Pathogenic genes of Vibrio vulnificusand protection by vaccination with an attenuated strain against the infection in mice (2011)
  • Author(s): Yamamoto, K., Yamamoto,M., Xiao, J., Nakata, K., K. Yokota, and K. Oguma.
  • Organizer: 日米医学協力研究会コレラ・細菌性腸管感染症専門部会・日本側総会.
  • Place of Presentation: 京都
  • Year and Date: 2011-08-03
• [Presentation] Vibrios as a causative agent of foodborn diseases (2010)
  • Author(s): Yamamoto, K.
  • Organizer: The First Guangzhou International Symposium on Nutrition and Health
  • Place of Presentation: Guangzhou, China
  • Year and Date: 2010-12-10
• [Presentation] (2010)
  • Author(s): Nakata, K., Tanaka, T., Yamamoto, K.,Soma, G.
  • Organizer: The 45th Annual Joint Panel Meeting on Cholera & Other Bacterial Enteric Infections.
  • Place of Presentation: Kyoto
  • Year and Date: 2010-12-06
• [Presentation] 腸管病原細菌に対する腸管マクロファージの応答性 (2010)
  • Author(s): 中田和江, 田中友美, 山本耕一郎, 杣源一郎
  • Organizer: 第63回日本細菌学会中国・四国支部総会
  • Place of Presentation: 松山
  • Year and Date: 2010-09-12
• [Presentation] Anti-stress effect by innate immunity activation (2010)
  • Author(s): Nakata, K., Yamamoto, M., Yamamoto, Inagawa, H., Soma, G.
  • Organizer: Bioactive Okayama 2010 - An international Conference for Biologically Active Substances.
  • Place of Presentation: Soja
  • Year and Date: 2010-08-12
• [Presentation] Vaccination with an attenuated Vibrio vulnificus strains protects mice against infection (2010)
  • Author(s): Yamamoto, M., Xiao, J., Hosohara, K., Segawa, R., Inoue, M., Nakai,M.,Yokota, K., Oguma, K., Yamamoto, K.
  • Organizer: Bioactive Okayama 2010 - An international Conference for Biologically Active Substances.
  • Place of Presentation: Soja
  • Year and Date: 2010-08-12
• [Presentation] 腸管マクロファージにおける LPS低応答性に関する研究.CD14 発現抑制機構の解析 (2010)
  • Author(s): 中田和江,山本耕一郎,吉岡典子,杣源一郎
  • Organizer: 日本細菌学雑誌,65:182,2010.日本細菌学会第83回総会
  • Place of Presentation: 横浜
  • Year and Date: 2010-03-28
• [Presentation] Vibrio vulnificus トランスポゾン挿入弱毒化変異株と挿入部位遺伝子の特定 (2010)
  • Author(s): 山本真衣,北林奏,肖建波,瀬川理恵,井上美幸,細原浩平,横田憲治,小熊恵二,山本耕一郎
  • Organizer: 日本細菌学雑誌, 65:150,2010. (日本細菌学会第83回総会
  • Place of Presentation: 横浜
  • Year and Date: 2010-03-27