Study of Shiga toxin productions us ing in vivo analysis

• Project/Area Number: 21590478
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bacteriology (including Mycology)
• Research Institution: Chiba University
• Principal Investigator: SHIMIZU Takeshi 千葉大学, 大学院・医学研究院, 准教授 (70312840)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2011: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2010: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2009: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: 腸管出血性大腸菌 / 志賀毒素 / 毒素産生 / 病原性 / NO / NO reductase / 感染症 / 細菌 / 分泌 / 細菌毒素 / 遺伝子発現

Research Abstract

A novel chromosome-plasmid hybrid bioluminescent reporter system (C-P reporter system) utilizing Photorhabdus luminescens luxCDABE genes has been constructed to monitor the expression of Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2) in enterohaemorrhagic Escherichia coli (EHEC) in real time. Using these reporter strains, we examined the profiles of Stx1 and Stx2 expression in EHEC. We found that production of both Stx1 and Stx2 in EHEC was enhanced upon contact with intestinal epithelial cells and within macrophages.

Research Products

• [Journal Article] Characterization of intracellular localization of PrP^<Sc> in prioninfected cells using monoclonal antibody that recognizes the region consisting of amino acids 119-127 of mouse PrP (2012)
  • Author(s): Yamasaki T., Suzuki A., Shimizu T., Watarai M., Hasebe R., Horiuchi M.
  • Journal Title: Journal of General Virology Volume: vol.93 no.3 Issue: 3 Pages: 668-680
  • DOI: 10.1099/vir.0.037101-0
  • Peer Reviewed
• [Journal Article] Recombinant Shiga toxin B subunit can induce neutralizing IgY antibody (2012)
  • Author(s): Paola Neri, Shunji Tokoro, Tsuyoshi Sugiyama, Kouji Umeda, Takeshi Shimizu, Takao Tsuji, Yoshikatsu Kodama, Hiroshi Mori
  • Journal Title: Biol.Pharm.Bull. Volume: (In press)
  • Peer Reviewed
• [Journal Article] Specific egg yolk immunoglobulin as a new preventive approach for Shiga-toxin-mediated diseases (2011)
  • Author(s): Paola Neri, Shunji Tokoro, Ryo Kobayashi, Tsuyoshi Sugiyama, Kouji Umeda, Takeshi Shimizu, Takao Tsuji, Yoshikatsu Kodama, Keiji Oguma Hiroshi Mori
  • Journal Title: PLoS ONE Volume: 6(10) Issue: 10 Pages: e26526-e26526
  • DOI: 10.1371/journal.pone.0026526
  • Peer Reviewed
• [Journal Article] Construction of a novel bioluminescent reportersystem for investigating Shiga toxin expression of enterohaemorrhagic (2011)
  • Author(s): Takeshi Shimizu, Yuko Ohta, Hiroyasu Tsutsuki Masatoshi Noda
  • Journal Title: Escherichia coli. Gene Volume: 478 Issue: 1-2 Pages: 1-10
  • DOI: 10.1016/j.gene.2011.01.006
  • Peer Reviewed
• [Journal Article] Construction of a novel bioluminescent reporter system for investigating Shiga toxin expression of enterohaemorrhagic.Escbericbia coli (2011)
  • Author(s): Takeshi Shimizu
  • Journal Title: Gene Volume: in press
  • Peer Reviewed
• [Journal Article] Shiga toxin 2 is specifically released from bacterial cells by two different mechanisms (2009)
  • Author(s): Takeshi Shimizu, Yuko Ohta Masatoshi Noda,
  • Journal Title: Infect. Immun Volume: 77 Issue: 7 Pages: 2813-2823
  • DOI: 10.1128/iai.00060-09
  • Peer Reviewed
• [Journal Article] Differential entry of Botulinum neurotoxin A into neural andintestinal cells (2009)
  • Author(s): Aurelie Couesnon, Takeshi Shimizu Michel R. Popoff
  • Journal Title: Cell. Microbiol Volume: 11 Issue: 2 Pages: 289-308
  • DOI: 10.1111/j.1462-5822.2008.01253.x
  • Peer Reviewed
• [Journal Article] Shiga toxin 2 is specifically released from bacterial cells by two different mechanisms. (2009)
  • Author(s): Takeshi Shimizu
  • Journal Title: Infect.Immun. 77 Pages: 2813-2823
  • Peer Reviewed
• [Journal Article] Yoshikatsu Kodama and Hiroshi Mori, 2012. Recombinant Shiga toxin B subunit can induce neutralizing IgY antibody
  • Author(s): Paola Neri, Shunji Tokoro, Tsuyoshi Sugiyama, Kouji Umeda, Takeshi Shimizu, Takao Tsuji
  • Journal Title: Pharm. Bull Volume: (in press)
  • Peer Reviewed
• [Presentation] NO biosensor plasmid for investigating NO levels within macrophages during bacterial infection (2012)
  • Author(s): 清水健、津々木博康、野田公俊
  • Organizer: 第85回日本細菌学会総会
  • Place of Presentation: 長崎ブリックホール(長崎)
  • Year and Date: 2012-03-27
• [Presentation] NO biosensor plasmid for investigating NO levels within macrophages during bacterial infection (2012)
  • Author(s): 清水健, 津々木博康, 野田公俊
  • Organizer: 第85回日本細菌学会総会
  • Place of Presentation: 長崎、長崎ブリックホール
• [Presentation] The nitric oxide reductase of Enterohemorrahgic Escherichia coli playsan important role for the survival within macrophages (2011)
  • Author(s): 清水健、津々木博康、野田公俊
  • Organizer: International Union of Microbiological Societies 2011 Congress
  • Place of Presentation: 札幌コンベンションセンター(北海道)
  • Year and Date: 2011-09-10
• [Presentation] 腸管出血性大腸菌の産生する志賀毒素について (2011)
  • Author(s): 清水健
  • Organizer: 緊急セミナー:腸管出血性大腸菌の今
  • Place of Presentation: 東京、東京慈恵会医科大学(招待講演)
  • Year and Date: 2011-08-02
• [Presentation] 腸管出血性大腸菌NO reductaseは志賀毒素2の産生を増加させる (2011)
  • Author(s): 清水健、津々木博康、野田公俊
  • Organizer: 第58回トキシンシンポジウム
  • Place of Presentation: 順天堂大学(東京)(招待講演)
  • Year and Date: 2011-07-06
• [Presentation] Haruaki Nakaya and Masatoshi Noda The nitric oxide reductase of enterohemorrhagic Escherichia coliO157:H7 increases Shiga toxin 2 production andplays an important role in its infection (2011)
  • Author(s): Takeshi Shimizu, Hiroyasu Tsutsuki, Akio Matsumoto
  • Organizer: 46^<th> Joint Conference on Cholera and Other Bacterial Enteric Infections Panel
  • Place of Presentation: Kolkata, India
• [Presentation] The nitric oxide reductase of enterohemorrahgic Escherichia coli plays an important role for the survival within macrophages (2011)
  • Author(s): Takeshi Shimizu, Hiroyasu Tsutsuki Masatoshi Noda
  • Organizer: International Union of Microbiological Societies2011 Congress
  • Place of Presentation: Sapporo Convention Center, Sapporo
• [Presentation] 清水健腸管出血性大腸菌の産生する志賀毒素について (2011)
  • Author(s): 清水健
  • Organizer: 緊急セミナー:腸管出血性大腸菌の今
  • Place of Presentation: 東京、東京慈恵会医科大学
• [Presentation] 野田公俊腸管出血性大腸菌のNO reductaseは志賀毒素2の産生を増加させる (2011)
  • Author(s): 清水健, 津々木博康
  • Organizer: 第58回トキシンシンポジウムシンポジウム指名講演
  • Place of Presentation: 東京、順天堂大学
• [Presentation] The nitric oxide reductase of enterohemorrhagic Escherichia coli 0157 : H7 increases Shiga toxin 2 production and plays an important role in its infection (2011)
  • Author(s): Takeshi Shimizu, Hiroyasu Tsutsuki, Akio Matsumoto, Haruaki Nakaya, Masatoshi Noda
  • Organizer: 46^<th> US-Japan Cholera and Other Bacterial Enteric Infections Joint Panel Meeting
  • Place of Presentation: インド(招待講演)
• [Presentation] Adherent effects of enterohemorrahgic Escberichia coli to human intestinal epithelial cells on its Shiga toxins production (2010)
  • Author(s): Takeshi Shimizu
  • Organizer: 45^<th> Joint Conference on Cholera and Other Bacterial Enteric Infections Panel
  • Place of Presentation: Kyoto University, Kyoto
  • Year and Date: 2010-12-07
• [Presentation] 腸管出血性大腸菌の腸管上皮細胞への接着は病原因子の発現を増加させる (2010)
  • Author(s): 清水健
  • Organizer: 第57回トキシンシンポジウム シンポジウム
  • Place of Presentation: 長浜、長浜ロイヤルホテル 指名講演
  • Year and Date: 2010-07-15
• [Presentation] Adherent effects of enterohemorrahgic Escherichia coli to human intestinal epithelial cells on its Shiga toxins production (2010)
  • Author(s): Takeshi Shimizu Masatoshi Noda
  • Organizer: 45^<th> Joint Conference on Cholera and OtherBacterial Enteric Infections Panel
  • Place of Presentation: Kyoto University, Kyoto
• [Presentation] 腸管出血性大腸菌の腸管上皮細胞への接着は病原因子の発現を増加させる (2010)
  • Author(s): 清水健, 野田公俊
  • Organizer: 第57回トキシンシンポジウムシンポジウム指名講演
  • Place of Presentation: 長浜、長浜ロイヤルホテル
• [Presentation] 腸管上皮細胞に接着した腸管出血性大腸菌はその志賀毒素1の産生量を増加させる (2010)
  • Author(s): 清水健, 野田公俊
  • Organizer: 第83回日本細菌学会総会
  • Place of Presentation: 横浜、パシフィコ横浜
• [Presentation] 腸管出血性大腸菌が産生する志賀毒素の菌体外放出機構 (2009)
  • Author(s): 清水健
  • Organizer: 第92回日本細菌学会関東支部総会シンポジウム
  • Place of Presentation: 東京、東京医科歯科大学
• [Presentation] Effect of phage-inducing agent on the number of Stx2-expressed cells in Shiga toxin-producing Escherichia coli (2009)
  • Author(s): Takeshi Shimizu, Yuko Ohta Masatoshi Noda
  • Organizer: 44^<rd> Joint Conference on Cholera and OtherBacterial Enteric Infections Panel
  • Place of Presentation: San Diego, California
• [Presentation] 腸管出血性大腸菌が産生する志賀毒素2の菌体外放出機構の解析 (2009)
  • Author(s): 清水健, 野田公俊
  • Organizer: 第56回トキシンシンポジウムシンポジウム指名講演
  • Place of Presentation: 岐阜、アルモニーテラッセ
• [Presentation] A mechanism of Stx2 release from enterohemorrahgic Escherichia coli (2009)
  • Author(s): Takeshi Shimizu, Yuko Ohta Masatoshi Noda
  • Organizer: The 13^<th> International Conference on EmergingInfectious Diseases in the Pacific Rim
  • Place of Presentation: Kolkata, India
• [Presentation] Effect of phage-inducing agent on the number of Stx2-expressed cells in Shiga toxin-producing Escherichia coli. (2009)
  • Author(s): Takeshi Shimizu
  • Organizer: 44^<rd> Joint Conference on Cholera and Other Bacterial Enteric Infections Panel
  • Place of Presentation: San Diego, California
• [Presentation] A mechanism of Stx2 release from enterohemorrahgic Escherichia coli (2009)
  • Author(s): Takeshi Shimizu
  • Organizer: The 13^<th> International Conference on Emerging Infectious Diseases in the Pacific Rim
  • Place of Presentation: Kolkata, India
• [Remarks]
  • URL: http://www.m.chiba-u.ac.jp/class/molecular-infectiology/indexMI.htm
• [Patent(Industrial Property Rights)] 一酸化窒素を探知するための形質転換用組換えベクター、およびこれを用いた一酸化窒素センサ細胞 (2011)
  • Inventor(s): 清水健, 津々木博康, 野田公俊
  • Industrial Property Rights Holder: 国立大学法人千葉大学
  • Industrial Property Number: 2011-101623
  • Filing Date: 2011-04-28