Development of prevention and treatment methods for infection by enterohemorrhagic Escherichia coli O157 based on bio-protect ive mechanisms

• Project/Area Number: 22590256
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General pharmacology
• Research Institution: University of Occupational and Environmental Health, Japan
• Principal Investigator: KOBAYSHI Hideyuki 産業医科大学, 医学部, 教授 (40148953)
• Co-Investigator(Kenkyū-buntansha): MORII Hiroyuki 産業医科大学, 医学部, 准教授 (60141743) ; FUJII Jun 鳥取大学, 医学部, 教授 (60271441) ; 大原 直也 岡山大学, 医歯薬総合研究科, 教授 (70223930)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2012: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2011: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2010: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 腸管出血性大腸菌 / ベロ毒素 / 感染症 / ワクチン / BCG / 遺伝子組み換え / 生理活性ペプチド / α抗原 / 生体防御機構 / 脳微小血管

Research Abstract

We have developed a novel vaccine using a recombinant BCG expressing the verotoxin B-subunit. The immunized mice with this vaccine survived longer than the nonvaccinated mice, confirminging the validity of this vaccine.
On the other hand, the expression of various bioactive peptides in the brain microvascular endothelial cells was changed by verotoxin. The expression of cytokines was increased suggesting that the interaction between brain microvascular endothelial cells and the leukocytes was changed by verotoxin.

Research Products

• [Journal Article] Investigation of Encephalopathy Caused by Shiga Toxin 2c-Producing Escheriohia ooli Infection in Mice (2013)
  • Author(s): Amran M. Y., Fujii J., Suzuki S. 0., Kolling G. L.,Villanueva A. M.,Kainunla M., Kobayashi H., Kanleyanla H. and Yoshida S
  • Journal Title: PLoS One Volume: 8 Issue: 3 Pages: e58959-e58959
  • DOI: 10.1371/journal.pone.0058959
  • Peer Reviewed
• [Journal Article] Antigen 85A and mycobacterial DNA-binding protein 1 are targets of immunoglobulin G in individuals with past tuberculosis (2013)
  • Author(s): Osada-Oka M, Tateishi Y, Hirayama Y, Ozeki Y, Niki M, Kitada S, Maekura R, Tsujimura K, Koide Y, Ohara N, Yamamoto T, Kobayashi K, Matsumoto S
  • Journal Title: Microbiol. Immunol Volume: 57 Pages: 30-37
  • Peer Reviewed
• [Journal Article] Antigen 85A and mycobacterial DNA-binding protein 1 are targets of immunoglobulin G in individuals with past tuberculosis. (2013)
  • Author(s): Osada-Oka M., Tateishi Y., Hirayama Y., Ozeki Y., Niki M., Kitada S., Maekura R., Tsujimura K., Koide Y., Ohara N., Yamamoto T., Kobayashi K., Matsumoto S.
  • Journal Title: Microbiol. Immunol. Volume: 57 Pages: 30-37
  • Peer Reviewed
• [Journal Article] Protection by recombinant Bacillus Calmette-Guerin vaccine expression Shiga toxin 2B subunit against Shiga toxin producing Escherichia coli in mice (2012)
  • Author(s): Fujii J., Naito M., Yutsudo T., Heatherly D., Yamada T., Kobayashi H., Yoshida S. and Obrig T
  • Journal Title: Clin. Vac. Imnunol Volume: 19 Issue: 12 Pages: 1932-1937
  • DOI: 10.1128/cvi.00473-12
  • Peer Reviewed
• [Journal Article] Current progress in tuberculosis and recombinant BCG vaccines (2012)
  • Author(s): Ohara N
  • Journal Title: J. Oral. Biosci Volume: 54 Pages: 92-95
  • Peer Reviewed
• [Journal Article] Current progress in tuberculosis and recombinant BCG vaccines. (2012)
  • Author(s): Ohara N.
  • Journal Title: J. Oral. Biosci. Volume: 54 Pages: 92-95
  • Peer Reviewed
• [Journal Article] Lipidphenotype of two distinct subpopulations of Vayo ohaoteri um bovis Bacillus Calmette-Guerin Tokyo 172 substrain (2011)
  • Author(s): Naka T., Maeda S., Niki M., Ohara N., Yamamoto S.,Yano I.,Maeyama J.,Ogura H., Kobayashi K.,Fujiwara N
  • Journal Title: Biol. Chem Volume: 286 Pages: 44153-44161
  • Peer Reviewed
• [Journal Article] Lipid phenotype of two distinct subpopulations of Mycobacterium bovis Bacillus Calmette-Guerin Tokyo 172 substrain (2011)
  • Author(s): Naka T., Maeda S., Niki M., Ohara N, Yamamoto S., Yano I., Maeyama J., Ogura H., Kobayashi K., Fujiwara N.
  • Journal Title: J Biol Chem Volume: 286 Pages: 44153-44161
  • Peer Reviewed
• [Journal Article] BCGワクチンの新たな活用 (2011)
  • Author(s): 大原直也
  • Journal Title: 岡山歯誌 Volume: 30 Pages: 51-60
  • Peer Reviewed
• [Journal Article] Suppressed induction of mycobacterial antigen-specific Th1-type CD4+Tcells in the lung after pulmonary mycobacterial infection (2010)
  • Author(s): Yahagi, A., Umemura, M., Tamura, T., Kariyone, A., Begum, M. D., Kawakami, K., Okamoto, Y., Hamada, S., Oshiro, K., Kohama, H., Arakawa, T., Ohara, N., Takatsu, K., Matsuzaki, G
  • Journal Title: Int. Immunol Volume: 22 Issue: 4 Pages: 307-318
  • DOI: 10.1093/intimm/dxq010
  • Peer Reviewed
• [Journal Article] アグリン(キーワード解説) (2010)
  • Author(s): 小林英幸、森井宏幸
  • Journal Title: 日薬理誌 Volume: 135 Pages: 215-216
  • Peer Reviewed
• [Journal Article] アグリン (2010)
  • Author(s): 小林英幸, 森井宏幸
  • Journal Title: 日薬理誌 Volume: 135 Pages: 215-216
  • NAID: 10026439433
  • Peer Reviewed
• [Journal Article] Suppressed induction of mycobacterial antigen-specific Thl-type CD4+ T cells in the lung after pulmonary mycobacterial infection.. (2010)
  • Author(s): Yahagi, A., Umemura, M., Tamura, T., Kariyone, A., Beg-um, M.D., Kawakami, K., Okamoto, Y., Hamada, S., Oshiro, K., Kohama, H., Arakawa, T., Ohara, N., Takatsu, K., Matsuzaki, G.
  • Journal Title: Int.Immunol Volume: 22 Pages: 307-318
  • Peer Reviewed
• [Presentation] 結核菌/BCGのストレプトマイシン耐性・依存性に関する新たな知見 (2012)
  • Author(s): 佐藤法仁,阿戸学,黒田誠,山崎利雄,松村隆之,関塚剛史,中山真彰,井上哲圭,本田尚子,土田耕三,小林和夫,大原直也
  • Organizer: 第82回実験結核研究会
  • Place of Presentation: 広島国際会議場、広島
  • Year and Date: 2012-05-09
• [Presentation] 抗酸菌のslidingmotilityと非貧食細胞への取り込みの関連性 (2012)
  • Author(s): 岡部真裕子,大原直也,藤原永年,中崇,阿戸学,小林和夫
  • Organizer: 第82回実験結核研究会
  • Place of Presentation: 広島国際会議場、広島
  • Year and Date: 2012-05-09
• [Presentation] マウスモデルを使った腸管出血性大腸菌感染O157による脳症発症メカニズムの解析 (2012)
  • Author(s): 藤井潤、Amran MY、吉田真一
  • 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] 腸管出血性大腸菌感染症による急性脳症の発症機序・診断と治療法の展望、シンポジウム「腸管出血性大腸菌の現状-その病態と疫学-」 (2012)
  • Author(s): 藤井潤
  • Organizer: 平成23年度日本獣医師会獣医学術学会年次大会
  • Place of Presentation: 札幌コンベンションセンター(札幌)(招待講演)
  • Year and Date: 2012-02-03
• [Presentation] Risk factors for hemolytic uremic syndrome caused by Shiga toxin-producing Escherichia coli infection in Japan (2012)
  • Author(s): Fujii J., Mizoue T., Nakada Y.,Ugajin S.,Naya Y.,Nakamura T.,Saitoh K.,Maruyama Y., Tada Y., Okabe N. and Yoshida S
  • Organizer: 8^<th> International Symposium on Shiga toxin (Verocytotoxin)- producing Escherichia coli infection
  • Place of Presentation: RAI Forum Complex, Amsterdam, The Netherlands
• [Presentation] Effectiveness azithromycin and/or Chinese medicine in the early and late stage of Shiga toxin-producing Escherichia coli-infected mouse models (2012)
  • Author(s): Amran MY, Fujii J, Yoshida S
  • Organizer: 8^<th> International Symposiunl on Shiga toxin (Verocytotoxin)-producing Escherichia coli infection
  • Place of Presentation: RAI Forum Complex, Amsterdam, The Netherlands
• [Presentation] ストレプトマイシン依存性結核菌18bの依存性に関与する遺伝子変異の解明と新たなストレプトマイシン耐性を誘導する変異の発見 (2012)
  • Author(s): 佐藤法仁,山崎利雄,小林和夫,大原直也
  • Organizer: 第87回日本結核病学会総会
  • Place of Presentation: 広島国際会議場、広島
• [Presentation] BCG Tokyo type I, II株の形態及び脂質分子の分布と機能 (2012)
  • Author(s): 藤原永年,中崇,前田伸司,柴田満,仁木満美子,大原直也,前山順一,矢野郁也,山本三郎
  • Organizer: 第87回日本結核病学会総会
  • Place of Presentation: 広島国際会議場、広島
• [Presentation] Mycobacterium avium 亜種 (avium・hominissuis)間での酸性環境下における菌体外pH上昇に関与するアンモニア産生経路の研究 (2012)
  • Author(s): 花村菜.月,堀田康弘,小川賢二,八木哲也,西森敬,大原直也,藤原永年,前田伸司,山崎利雄,伊藤佐生智,瀧井猛将
  • Organizer: 第87回日本結核病学会総会
  • Place of Presentation: 広島国際会議場、広島
• [Presentation] Lipid Phenotypes of Two Distinct Subpopulations of Mycobacterium bovis Bacillus Calnlette-Guerin Tokyo 172 Substrain and their Host Responses (2012)
  • Author(s): Naka, T.,Maeda S.,Niki M., Ohara N.,Yamamoto S.,Yano I.,Maeyama J.,Ogura H.,Kobayashi K., Shibata M., and Fujiwara N
  • Organizer: American Society for Microbiology 112th General Meeting
  • Place of Presentation: Moscone Center, San Francisco, U.S.A
• [Presentation] Investigation of acute encephalopathy caused by Shiga toxin 2c-producing enterohemorrhagic E. coli 0157 in mice. (2012)
  • Author(s): Fujii J, Amran MY, Yoshida S
  • Organizer: U.S.-Japan cooperarive medical science program
  • Place of Presentation: Chiba University, Chiba Japan
• [Presentation] Risk factors for hemolytic uremic syndrome caused by Shiga toxin-producing Escherichia coli infection in Japan. (2012)
  • Author(s): Fujii J., Mizoue T., Nakada Y., Ugajin S., Naya Y., Nakamura T., Saitoh K., Maruyama Y., TadaY., Okabe N., Yoshida S.
  • Organizer: 8th International Symposium on Shiga toxin (Verocytotoxin)- producing Escherichia coli infection
  • Place of Presentation: RAI Forum Complex, Amsterdam, The Netherlands
• [Presentation] Effectiveness azithromycin and/or Chinese medicine in the early and late stage of Shiga toxin-producing Escherichia coli-infected mouse models. (2012)
  • Author(s): Amran M.Y., Fujii J., Yoshida S.
  • Organizer: 8th International Symposium on Shiga toxin (Verocytotoxin)-producing Escherichia coli infection
  • Place of Presentation: RAI Forum Complex, Amsterdam, The Netherlands
• [Presentation] 結核菌／BCGのストレプトマイシン耐性・依存性に関する新たな知見． (2012)
  • Author(s): 佐藤法仁，阿戸学，黒田誠，山崎利雄，松村隆之，関塚剛史，中山真彰，井上哲圭，本田尚子，土田耕三，小林和夫，大原直也
  • Organizer: 第82回実験結核研究会
  • Place of Presentation: 広島国際会議場、広島
• [Presentation] 抗酸菌のsliding motilityと非貪食細胞への取り込みの関連性． (2012)
  • Author(s): 岡部真裕子，大原直也，藤原永年，中崇，阿戸学，小林和夫
  • Organizer: 第82回実験結核研究会
  • Place of Presentation: 広島国際会議場、広島
• [Presentation] ストレプトマイシン依存性結核菌18bの依存性に関与する遺伝子変異の解明と新たなストレプトマイシン耐性を誘導する変異の発見． (2012)
  • Author(s): 佐藤法仁，山崎利雄，小林和夫，大原直也
  • Organizer: 第87回日本結核病学会総会
  • Place of Presentation: 広島国際会議場、広島
• [Presentation] BCG Tokyo type I, II株の形態及び脂質分子の分布と機能． (2012)
  • Author(s): 藤原永年，中崇，前田伸司，柴田満，仁木満美子，大原直也，前山順一，矢野郁也，山本三郎
  • Organizer: 第87回日本結核病学会総会
  • Place of Presentation: 広島国際会議場、広島
• [Presentation] Mycobacterium avium亜種（avium・hominissuis）間での酸性環境下における菌体外pH上昇に関与するアンモニア産生経路の研究． (2012)
  • Author(s): 花村菜月，堀田康弘，小川賢二，八木哲也，西森敬，大原直也，藤原永年，前田伸司，山崎利雄，伊藤佐生智，瀧井猛将
  • Organizer: 第87回日本結核病学会総会
  • Place of Presentation: 広島国際会議場、広島
• [Presentation] Lipid Phenotypes of Two Distinct Subpopulations of Mycobacterium bovis Bacillus Calmette-Guerin Tokyo 172 Substrain and their Host Responses. (2012)
  • Author(s): Naka T., Maeda S., Niki M., Ohara N., Yamamoto S., Yano I., Maeyama J., Ogura H., Kobayashi K., Shibata M., Fujiwara N.
  • Organizer: American Society for Microbiology 112th General Meeting
  • Place of Presentation: Moscone Center, San Francisco，U.S.A.
• [Presentation] Investigation of acute encephalopathy caused by Shiga toxin 2c-producing enterohemorrhagic E. coli O157 in mice. (2012)
  • Author(s): Fujii J., Amran M.Y., Yoshida S.
  • Organizer: U.S.-Japan cooperarive medical science program
  • Place of Presentation: Chiba University, Chiba
• [Presentation] Analysis of the 16S rRNA gene of the streptomycin-dependent Mycobacterium tuberculosis strain 18b and phenolypic revenant strains (2011)
  • Author(s): Sato N., Ato M., Yamazaki T., Kuroda M., Matsumura T., Sekizuka T., Nakayama M., Honda, N., Tsuchida, K., Kobayashi K., Ohara N
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜)
  • Year and Date: 2011-12-16
• [Presentation] Successful azithromycin and Daiou combination treated of mouse model orally infected with Shiga toxin-producing Escherichia coli (2011)
  • Author(s): Amran MY, Fujii J, Yoshida S
  • Organizer: 46^<nd> Annual joint panel meeting, Cholera and other bacterial enteric infections, U.S.-Japan cooperarive medical science program
  • Place of Presentation: Hyatt Regency Hotel (Kolkata)
  • Year and Date: 2011-12-12
• [Presentation] Streptomycin依存性結核菌における遺伝子変異の解明とStreptomycin耐性を誘導する新たな遺伝子変異 (2011)
  • Author(s): 佐藤法仁、阿戸学、松村隆之、山崎利雄、関塚剛史、黒田誠、中山真章、井上哲圭、本田尚子、土田耕三、小林和夫、大原直也
  • Organizer: 第64回日本細菌学会中国・四国支部総会
  • Place of Presentation: 岡山大学(岡山)
  • Year and Date: 2011-10-22
• [Presentation] Proteomic analysis of urine of rats infected with Leptospira interogans serovar Manilae (2011)
  • Author(s): Segawa T, Nomura K, Villanueva SYAM, Fujii J, Yoshida S
  • Organizer: International Leptosirosis Society
  • Place of Presentation: Unversidad AutOnoma de Yucatan (Yucatan)
  • Year and Date: 2011-09-19
• [Presentation] Successful azithromycin treatment of a fatal neurological mouse model orally infected with and Shiga toxin 2c-producing Escherichia coli O157:H (2011)
  • Author(s): Amran MY, Fujii J., Yoshida S
  • Organizer: International Union of Microbiological Societies 2011 Congress
  • Place of Presentation: 札幌コンベンションセンター(札幌)
  • Year and Date: 2011-09-10
• [Presentation] Proteomic analysis of urine of rats infected with Leptospira interogans serovar Manilae (2011)
  • Author(s): Segawa T., Nomura K., Villanueva SYA, Nomura K, Fujii J, Yoshida S
  • Organizer: International Union of Microbiological Societies 2011 Congress
  • Place of Presentation: 札幌コンベンションセンター(札幌)
  • Year and Date: 2011-09-10
• [Presentation] Identification of the novel mutations in 16S rRNA gene of the substrains of a streptomycin-dependent Mycobacterium tuberculosis Strain18b which confer streptomycin resistance in Mycobacterium (2011)
  • Author(s): Sato N., Ato M., Matsumura T., Yamazaki T., Sekizuka T., Kuroda M., Nakayama M., Kobayashi K., Ohara N
  • Organizer: XIII International Congress of Bacteriology and Applied Microbiology. International Union of Microbiological Societies
  • Place of Presentation: 札幌コンベンションセンター(札幌)
  • Year and Date: 2011-09-10
• [Presentation] Gene expression profile of in a thymidine synthase ThyX-deletion mutant of Mycobacterium bovis BCG (2011)
  • Author(s): Ohara N., Satoh N., Nakayama M., Okabe M, Kobayashi K
  • Organizer: XIII International Congress of Bacteriology and Applied Microbiology. International Union of Microbiological Societies
  • Place of Presentation: 札幌コンベンションセンター(札幌)
  • Year and Date: 2011-09-10
• [Presentation] わが国において成人女性は腸管出血性大腸菌感染による溶血性尿毒症症候群のリスク要因である (2011)
  • Author(s): 藤井潤、溝上哲也、北敏郎、城宏輔、岸本宏志、名矢結香、中村勤、中田友里、宇賀神貞夫、斎藤京子、丸山保夫、多田有希、岡部信彦、吉田真一
  • Organizer: 第15回腸管出血性大腸菌感染症研究会
  • Place of Presentation: 北浜フォーラム(大阪)
  • Year and Date: 2011-07-15
• [Presentation] 抗酸菌におけるセカンドメッセンジャーの利用 (2011)
  • Author(s): 佐藤法仁、中山真彰, 岡部真裕子, 小林和夫、大原直也
  • Organizer: 第86回実験結核研究会
  • Place of Presentation: 大妻講堂(東京)
  • Year and Date: 2011-06-01
• [Presentation] 抗酸菌のチミジル酸合成酵素の研究と新たなBCGワクチン作製の試み (2010)
  • Author(s): 大原直也
  • Organizer: 第690回獣医学研究談話会
  • Place of Presentation: 北海道大学(札幌)
  • Year and Date: 2010-10-14
• [Presentation] マイコバクテリアのチミジル酸合成酵素と組換えBCGワクチンシンポジウム口腔と関連器官の感染症5W1Hなぜわたしはこの研究をしているのか (2010)
  • Author(s): 大原直也
  • Organizer: 第52回歯科基礎医学会学術大会
  • Place of Presentation: タワーホール船堀(東京)
  • Year and Date: 2010-09-22
• [Presentation] Analysis of RD16 locus in the BCG Tokyo 172 genome.Forty-fifth Joint Research Conference on Leprosy and Tuberculosis. (2010)
  • Author(s): Ohara, N., Okabe, M., Yamamoto, S., Takii, T., Fujiwara, N., Maeyama, J., Nakayama, M., Kobayashi, K.
  • Organizer: The United State-Japan Cooperative Medical Science Program
  • Place of Presentation: Broad Institute (Cambridge, Massachusetts, U.S.A.)
  • Year and Date: 2010-07-15
• [Presentation] Isolation and characterization of a rpoZ mutant of Mycobacterium smegmatis.Forty-fifth Joint Research Conference on Leprosy and Tuberculosis. (2010)
  • Author(s): Taniguchi, H., Nomoto, M., Ogawa, M., Fukuda, K., Ohara, N., Fujiwara, N.
  • Organizer: The United State-Japan Cooperative Medical Science Program
  • Place of Presentation: Broad Institute (Cambridge, Massachusetts, U.S.A.)
  • Year and Date: 2010-07-14
• [Presentation] 形態の異なるBCG Tokyo 172亜種間の脂質生化学的比較 (2010)
  • Author(s): 藤原永年, 前田伸司, 吉村満美子, 大原直也, 前山順一, 瀧井猛将, 矢野郁也, 山本三郎
  • Organizer: 第85回日本結核病学会総会
  • Place of Presentation: 京都テルサ(京都)
  • Year and Date: 2010-05-20
• [Presentation] BCG Tokyo 172株に存在するサブポピュレーション間の遺伝子発現の比較発現の比較検討 (2010)
  • Author(s): 大原直也, 岡部真裕子, 山本三郎, 瀧井猛将, 藤原永年, 前山順一, 佐藤法仁, 井上哲圭, 小林和夫
  • Organizer: 第80回実験結核研究会
  • Place of Presentation: 京都テルサ(京都)
  • Year and Date: 2010-05-19
• [Presentation] DNAマイクロアレイを用いたBCG Tokyo 172-1に存在するサブポピュレーションの遺伝子発現の比較検討 (2010)
  • Author(s): 大原直也, 山本三郎, 瀧井猛将, 藤原永年, 前山順一, 小林和夫
  • Organizer: 第80回実験結核研究会
  • Place of Presentation: 京都テルサ(京都)
  • Year and Date: 2010-05-19