Research on the fish cytosolic LPS sensor and its mechanism

• Project/Area Number: 26450288
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Aquatic life science
• Research Institution: University of Miyazaki
• Principal Investigator: Hikima Junichi 宮崎大学, 農学部, 准教授 (70708130)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: トラフグ / NLR / カスパーセ１ / インフラマソーム / メダカ / LPS / NLRファミリー遺伝子 / 炎症反応 / 炎症性サイトカイン遺伝子 / パターン認識 / 魚類 / NOD様受容体（NLR）

Outline of Final Research Achievements

To discover the first sensor to recognize LPS in the cytosol, the genes consisted of inflammasome were cloned in Fugu (Takifugu rubripes). These genes were significantly induced in the kidney cells stimulated with nigericin, silica and alum. Furthermore, medaka (Oryzias latipes) NLR-PYD and Caspase-1 genes were also cloned and the caspase-1 activity were demonstrated and confirmed. These suggest that there is probably an inflammasome-like activation system in fish.

Research Products

• [Journal Article] Complete Genome Sequence of Photobacterium damselae subsp. piscicida Strain OT-51443 Isolated from Yellowtail (Seriola quinqueradiata) in Japan (2017)
  • Author(s): Aoki T, Teru Y, Kono T, Sakai M , Takano T, Hawke JP, Fukuda Y, Takeyama H, Hikima J
  • Journal Title: Genome Announcements Volume: 印刷中
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Direct Detection of Astaxanthin from Salmon Filet Using Raman Spectroscopy (2017)
  • Author(s): Hikima J, Ando M, Hamaguchi H, Sakai M, Maita M, Yazawa K, Takeyama H, Aoki T
  • Journal Title: Marine Biotechnology Volume: 19 Issue: 2 Pages: 157-163
  • DOI: 10.1007/s10126-017-9739-7
  • Peer Reviewed / Open Access
• [Journal Article] Molecular Characterization and Expression Analysis of Tumor Necrosis Factor Alpha-induced Protein 3 (TNFAIP3/A20) Gene from Japanese Pufferfish <i>Takifugu rubripes</i> (2017)
  • Author(s): Hikima J, Morita M, Kinoshita S, Basu M, Biswas G, Kono T, Sakai M
  • Journal Title: Fish Pathology Volume: 52 Issue: 1 Pages: 15-22
  • DOI: 10.3147/jsfp.52.15
  • NAID: 130005548817
  • ISSN: 0388-788X, 1881-7335
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Characterization of a specific monoclonal antibody against immunoglobulin light kappa/L1 chain in olive flounder (Paralichthys olivaceus) (2017)
  • Author(s): Kim YK, Lee JS, Jung JW, Hikima J, Ohtani M, Jang HB, Nho SW, Cha IS, Park SB, Lee JH, Aoki T, Jung TS
  • Journal Title: Fish and Shellfish Immunology Volume: 60 Pages: 88-96
  • DOI: 10.1016/j.fsi.2016.11.023
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Detection of Kudoa septempunctata in the Japanese Flounder by Loop-Mediated Isothermal Amplification (LAMP) Targeting the Internal Transcribed Spacer (ITS) Regions (2016)
  • Author(s): Biswas G, Kodama H, Mekata T, Kono T, Yokoyama H, Sakai M, Hikima J
  • Journal Title: Turkish Journal of Fisheries and Aquatic Sciences Volume: 16 Issue: 3 Pages: 623-628
  • DOI: 10.4194/1303-2712-v16_3_15
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Inflammatory immune response by lipopolysaccharide-responsive nucleotide binding oligomerization domain (NOD)-like receptors in the Japanese pufferfish (Takifugu rubripes). (2016)
  • Author(s): Biswas G, Bilen S, Kono T, Sakai M, Hikima J
  • Journal Title: Developmental and Comparative Immunology Volume: 55 Pages: 21-31
  • DOI: 10.1016/j.dci.2015.10.008
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Expression and biological activity of two types of interferon genes in medaka (Oryzias latipes). (2016)
  • Author(s): Maekawa S, Chiang YA, Hikima J, Sakai M, Lo CF, Wang HC, Aoki T
  • Journal Title: Fish and Shellfish Immunology Volume: 48 Pages: 20-29
  • DOI: 10.1016/j.fsi.2015.11.036
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Immune responses in the Japanese pufferfish (Takifugu rubripes) head kidney cells stimulated with particulate silica. (2016)
  • Author(s): Morimoto T, Biswas G, Kono T, Sakai M, Hikima J
  • Journal Title: Fish and Shellfish Immunology Volume: 49 Pages: 84-90
  • DOI: 10.1016/j.fsi.2015.12.023
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Inductive immune responses in the Japanese pufferfish (Takifugu rubripes) treated with recombinant IFN-γ, IFN-γrel, IL-4/13A and IL-4/13B (2016)
  • Author(s): Gouranga Biswas, Ryusuke Nagamine, Jun-ichi Hikima, Masahiro Sakai, Tomoya Kono
  • Journal Title: International Immunopharmacology Volume: 31 Pages: 50-56
  • DOI: 10.1016/j.intimp.2015.12.005
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] SPFクルマエビにおける高感度選抜法の開発．養殖技術講座&#8212;SPFクルマエビ種苗－第１回． (2016)
  • Author(s): 引間順一・浜野かおる
  • Journal Title: 月刊養殖ビジネス Volume: 1 Pages: 19-22
• [Journal Article] Draft Genome Sequence of the Fish Pathogen Mycobacterium pseudoshottsii Strain JCM15466, a Species Closely Related to M. marinum. (2016)
  • Author(s): Hikima J, Sakai M, Aoki T, Takeyama H, Hawke J, Mori K, Tashiro K, Kuhara S.
  • Journal Title: Genome Announcements Volume: 4 Issue: 1
  • DOI: 10.1128/genomea.01630-15
  • Open Access / Int'l Joint Research
• [Journal Article] Detection of <i>Kudoa amamiensis</i> Using Loop-Mediated Isothermal Amplification (LAMP) (2015)
  • Author(s): Sakai M, Biswas G, Kono T, Hikima J, Yokoyama H.
  • Journal Title: Fish Pathology Volume: 50 Issue: 3 Pages: 119-122
  • DOI: 10.3147/jsfp.50.119
  • NAID: 130005100742
  • ISSN: 0388-788X, 1881-7335
  • Peer Reviewed / Open Access
• [Journal Article] DNA vaccine-mediated innate immune response triggered by PRRs in teleosts (2015)
  • Author(s): Aoki T, Takano T, Hikima J
  • Journal Title: Fisheries Science Volume: 81 Issue: 2 Pages: 205-217
  • DOI: 10.1007/s12562-014-0845-4
  • NAID: 40020416307
  • Peer Reviewed / Open Access
• [Journal Article] The cytosolic sensor, DDX41, activates antiviral and inflammatory immunity in response to stimulation with double-stranded DNA adherent cells of the olive flounder, Paralichthys olivaceus. (2015)
  • Author(s): Quynh NT, Hikima J, Kim YR, Fagutao FF, Jung TS, Aoki T
  • Journal Title: Fish and Shellfish Immunology Volume: 44 Issue: 2 Pages: 576-583
  • DOI: 10.1016/j.fsi.2015.03.008
  • Peer Reviewed
• [Journal Article] Infalammatory responses in the Japanese pufferfish (Takifugu rubripes) head kidney cells stimulated with an inflammasome-inducing agenet, nigericin (2014)
  • Author(s): S. Bilen, G. Biswas, S. Otsuyama, T. Kono, M. Sakai and J. Hikima
  • Journal Title: Developmental and Comparative Immunology Volume: 46 Issue: 2 Pages: 222-230
  • DOI: 10.1016/j.dci.2014.04.010
  • Peer Reviewed
• [Presentation] メダカ腸管におけるIL-17の役割を明らかにするためのIL-17A/F1変異体のトランスクリプトーム解析 (2017)
  • Author(s): 引間順一、池田大介、和泉幹久、森本和月、河野智哉、酒井正博、竹山春子、青木 宙、水澤奈々美、渡部終五、木下政人
  • Organizer: 平成２８年度日本水産学会春季大会
  • Place of Presentation: 東京海洋大学（東京、品川）
  • Year and Date: 2017-03-26
  • Int'l Joint Research
• [Presentation] A Molecular Adjuvant Based on the Pattern Recognition System Using Toll-Like Receptor 5 and Bacterial Flagellin Genes in Teleost Fish (2016)
  • Author(s): Natsuki Morimoto, Tomoya Kono, Masakazu Kondo, Masahiro Sakai, Jun-ichi Hikima
  • Organizer: The 10th IMT-GT UNINET Conference 2016
  • Place of Presentation: Prince of Songkla University (Hat Yai, Thailand)
  • Year and Date: 2016-12-01
  • Int'l Joint Research
• [Presentation] Establishment of ASC-knockout medaka using CRISPR/Cas system (2016)
  • Author(s): Nastuki Morimoto, Shun Maekawa, Han-Ching Wang, Takashi Aoki, Kono Tomoya, Masaniro Sakai, Jun-ichi Hikima
  • Organizer: he 22nd Japanese Medaka and Zebrafish Meeting (NBRP Medaka and Zebrafish Joint International Meeting)
  • Place of Presentation: NBRP (Okazaki, Aichi)
  • Year and Date: 2016-08-20
  • Int'l Joint Research
• [Presentation] Cytosolic CpGs induce DHX9 and DHX36 pathway mediated through MyD88 in teleosts (2016)
  • Author(s): Jun-ichi Hikima, Koji Ebisu, Tomoya Kono, Masahiro Sakai
  • Organizer: The 7th World Fisheries Congress in Busan
  • Place of Presentation: Busan Bexco (Busan, Korea)
  • Year and Date: 2016-05-23
  • Int'l Joint Research
• [Presentation] ラマン分光法によるマグロ筋肉に含まれる多価不飽和脂肪酸の検出の試み (2016)
  • Author(s): 引間順一・安藤正浩・仲西友紀・河原 聡・酒井正博・濵口宏夫・竹山春子・青木 宙
  • Organizer: 平成２８年度日本水産学会春季大会
  • Place of Presentation: 東京海洋大学（東京、品川）
  • Year and Date: 2016-03-26
• [Presentation] 遺伝子診断技術を用いた簡便なエビ種判別法の確立 (2016)
  • Author(s): 西 純市、岩崎直子、阪地英男、引間順一、酒井正博
  • Organizer: 平成２８年度日本水産学会春季大会
  • Place of Presentation: 東京海洋大学（東京、品川）
  • Year and Date: 2016-03-26
• [Presentation] 空気暴露ストレス負荷によるストレス応答遺伝子の発現パターンを利用したウイルスキャリアー親エビの選抜 (2016)
  • Author(s): 酒井正博・児玉ひろの・引間順一・河野智哉・伊丹利明
  • Organizer: 平成２８年度日本魚病学会春季大会
  • Place of Presentation: 日本獣医生命科学大学（東京、武蔵野）
  • Year and Date: 2016-03-12
• [Presentation] アラムを接種したトラフグにおけるサイトカイン遺伝子およびインフラマソーム関連遺伝子の発現動態 (2016)
  • Author(s): 河野智哉・森本隆志・酒井正博・引間順一
  • Organizer: 平成２８年度日本魚病学会春季大会
  • Place of Presentation: 日本獣医生命科学大学（東京、武蔵野）
  • Year and Date: 2016-03-12
• [Presentation] 16SrRNA-based metagenomics analysis of microorganisms from cultured tilapia ponds in Thailand (2016)
  • Author(s): Aoki T, Watanabe U, Takeyama H, Hattori M, Suda W, Hikima J, Sakai M, Unajak S, Rapeepat M
  • Organizer: HNRCT-JSPS Asian Core Program Symposium 2016
  • Place of Presentation: Kasetsart University (Bangkok, Thailand)
  • Year and Date: 2016-02-12
  • Int'l Joint Research
• [Presentation] Cytosolic pattern recognition system in fish (2016)
  • Author(s): Jun-ichi Hikima
  • Organizer: The Third Symposium for Different Field Exchange by Tenure Track Researchers in Tokyo University of Marine Science and Technology
  • Place of Presentation: Tokyo University of Marine Science and Technology (Shinagawa, Tokyo)
  • Int'l Joint Research / Invited
• [Presentation] トラフグにおける粒子状結晶化物質の刺激に対する免疫応答 (2015)
  • Author(s): 森本隆志・河野智哉・酒井正博・引間順一
  • Organizer: 平成27年度日本水産学会九州支部大会
  • Place of Presentation: 宮崎大学（宮崎）
  • Year and Date: 2015-11-07
• [Presentation] 2種類のメダカI型インターフェロンによって誘導される免疫応答 (2015)
  • Author(s): 前川 峻・青木 宙・王 涵青・羅 竹芳・竹山春子・引間順一・酒井正博
  • Organizer: 平成２７年度日本魚病学会秋季大会
  • Place of Presentation: 東京大学（東京、文京）
  • Year and Date: 2015-09-25
• [Presentation] クルマエビにおけるストレス応答遺伝子の発現動態~特に低酸素負荷に対する応答~ (2015)
  • Author(s): 児玉ひろの・引間順一・河野智哉・稲田真理・伊丹利明・酒井正博
  • Organizer: 平成２７年度日本魚病学会秋季大会
  • Place of Presentation: 東京大学（東京、文京）
  • Year and Date: 2015-09-25
• [Presentation] 水産無脊椎動物であるエビ類・貝類のIL-6ホモログに関する研究 (2015)
  • Author(s): 稲田真理・米加田徹・引間順一・酒井正博・伊丹利明
  • Organizer: 平成２７年度日本魚病学会秋季大会
  • Place of Presentation: 東京大学（東京、文京）
  • Year and Date: 2015-09-25
• [Presentation] トラフグ頭腎細胞におけるDAMPs刺激に対する免疫応答 (2015)
  • Author(s): 引間順一・森本隆志・河野智哉・酒井正博
  • Organizer: 平成２７年度日本水産学会秋季大会
  • Place of Presentation: 東北大学（宮城、仙台）
  • Year and Date: 2015-09-22
• [Presentation] METAGENOMIC ANALYSES OF BACTERIAL FLORA IN THE POND WATER AND INTESTINE OF TILAPIA IN THAILAND (2015)
  • Author(s): T. Aoki, U. Watanabe, H. Takeyama, W. Suda, S. Hattori, J. Hikima, M. Sakai, S. Unajak, M. Rapeepat, HC Wang, CF Lo
  • Organizer: 17th International Conference on Diseases of Fish and Shellfish (EAFP conference)
  • Place of Presentation: Las Palmas de Grand Canaria, Spain
  • Year and Date: 2015-09-07
  • Int'l Joint Research
• [Presentation] メダカⅠ型インターフェロン遺伝子の同定と生物活性 (2015)
  • Author(s): 前川 峻，青木 宙，王 涵青，羅 竹芳，竹山 春子，引間 順一，酒井 正博
  • Organizer: 第27回日本比較免疫学会学術集会
  • Place of Presentation: 福井県立大学（福井、小浜
  • Year and Date: 2015-08-21
• [Presentation] Immune responses in the Japanese pufferfish (Takifugu rubripes) head kidney cells stimulated with particulate silica (2015)
  • Author(s): Masahiro Sakai, Takashi Morimoto, Gouranga Biswas, Tomoya Kono, Jun-ichi Hikima
  • Organizer: 13th International Society of Developmental & Comparative Immunology
  • Place of Presentation: Murcia, Spain
  • Year and Date: 2015-06-28
  • Int'l Joint Research
• [Presentation] Involvement of an intracellular pattern recognition receptor, NOD-like receptor in inflammatory response of the Japanese pufferfish (Takifugu rubripes) (2015)
  • Author(s): Jun-ichi Hikima, Gouranga Biswas, Tomoya Kono, Masahiro Sakai
  • Organizer: 13th International Society of Developmental & Comparative Immunology
  • Place of Presentation: Murcia, Spain
  • Year and Date: 2015-06-28
  • Int'l Joint Research
• [Presentation] Expression and biological activity of two types of interferon genes in medaka fish (Oryzias latipes) (2015)
  • Author(s): Shun Maekawa, Takashi Aoki, Han-Ching Wang, Yi-An Chiang, Chu-Fang Lo, Haruko Takeyama, Jun-Ichi Hikima, Masahiro Sakai
  • Organizer: 13th International Society of Developmental & Comparative Immunology
  • Place of Presentation: Murcia, Spain
  • Year and Date: 2015-06-28
  • Int'l Joint Research
• [Presentation] フグA20遺伝子の全長クローニングおよび発現解析 (2015)
  • Author(s): 酒井正博、森田美咲、森本隆司、河野智哉、引間順一
  • Organizer: 第17回マリンバイオテクノロジー学会
  • Place of Presentation: 東京海洋大学（東京、品川）
  • Year and Date: 2015-05-30
• [Presentation] フグDHX9および36遺伝子のクローン化およびCpGによる遺伝子発現誘導 (2015)
  • Author(s): 引間順一、戎 香治、林 可奈子、河野智哉、酒井正博
  • Organizer: 第17回マリンバイオテクノロジー学会
  • Place of Presentation: 東京海洋大学（東京、品川）
  • Year and Date: 2015-05-30
• [Presentation] リポ多糖に応答するフグNLRファミリー遺伝子の発現解析 (2015)
  • Author(s): 引間順一、Gouranga Biswas、河野智哉、酒井正博
  • Organizer: 平成27年度日本水産学会春季大会
  • Place of Presentation: 東京海洋大学（品川、東京）
  • Year and Date: 2015-03-27 – 2015-03-31
• [Presentation] Immunostimulant effects of recombinant cytokine administration in the Japanese pufferfish, Takifugu rubripes (2014)
  • Author(s): R. Nagamine, G. Biswas, J. Hikima, M. Sakai. T. Kono
  • Organizer: Seventh International Symposium on Aquatic Animal Health
  • Place of Presentation: Portland, OR, USA
  • Year and Date: 2014-08-31 – 2014-09-04
• [Presentation] Cytosolic sensor, DDX41 activates antiviral and inflammatory immunity in response to stimulation with dsDNA in Japanese flounder, Paralichthys olivaceus (2014)
  • Author(s): J. Hikima, N.T Quynh, F.F. Fagutao, M. Sakai, T.S. Jung, T. Aoki
  • Organizer: Seventh International Symposium on Aquatic Animal Health
  • Place of Presentation: Portland, OR, USA
  • Year and Date: 2014-08-31 – 2014-09-04
• [Presentation] A new strategy to characterize different cell types from fish leukocytes using the bio-imaging technology of the confocal Raman microspectroscopy (2014)
  • Author(s): T. Aoki, M. Ando, H. Hamaguchi, J. Hikima, M. Sakai, T. Moritomo, T. Nakanishi, H. Takeyama
  • Organizer: Seventh International Symposium on Aquatic Animal Health
  • Place of Presentation: Portland, OR, USA
  • Year and Date: 2014-08-31 – 2014-09-04
• [Presentation] Immunostimulant effects of the Mongolian dairy product derived lactic acid bacteria in the Japanese pufferfish (Takifugu rubripes) (2014)
  • Author(s): G. Biswas, T. Kono, J. Hikima, M. Sakai
  • Organizer: Seventh International Symposium on Aquatic Animal Health
  • Place of Presentation: Portland, OR, USA
  • Year and Date: 2014-08-31 – 2014-09-04
• [Presentation] Cytokine-induced CD4 positive T cell differentiation in the Japanese pufferfish, Takifugu rubripes (2014)
  • Author(s): T. Kono. H. Korenaga, R. Nagamine, G. Biswas, J. Hikima, M. Sakai
  • Organizer: Seventh International Symposium on Aquatic Animal Health
  • Place of Presentation: Portland, OR, USA
  • Year and Date: 2014-08-31 – 2014-09-04
• [Presentation] Inflammatory responses in head kidney cells stimulatied with as inflammasome inducer NIgericin in the Japanese pufferfish (Takifugu rubrepes) (2014)
  • Author(s): J. Hikima, G. Biswas, S. Bilen, S. Otsuyama, T. Kono, M. Sakai
  • Organizer: The 10th Asia-Pacific Marine Biotechnology Conference
  • Place of Presentation: Academia Sinica (Taipei, Taiwan)
  • Year and Date: 2014-05-04 – 2014-05-08
• [Presentation] Involvement of an intracellular multi-protein complex, inflammasome in the inflammatory immune response of the Japanese pufferfish (Takifugu rubripes) (2014)
  • Author(s): G. Biswas, T. Kono, M. Sakai, J. Hikima
  • Organizer: The 10th Asia-Pacific Marine Biotechnology Conference
  • Place of Presentation: Academia Sinica (Taipei, Taiwan)
  • Year and Date: 2014-05-04 – 2014-05-08
• [Presentation] In silico identification and expression analysis of tumor necrosis factor super family genes in the Japanese pufferfish, fugu (Takifugu rubripes) (2014)
  • Author(s): M. Sakai, S. Kinoshita, G. Biswas, J. Hikima, T. Kono
  • Organizer: The 10th Asia-Pacific Marine Biotechnology Conference
  • Place of Presentation: Academia Sinica (Taipei, Taiwan)
  • Year and Date: 2014-05-04 – 2014-05-08
• [Book] Prevention And Treatment Of Diseases, Section 2.2. Diagnosis &#8211; PCR Detection. In: Fish Diseases (Ed. Takashi Aoki) (2016)
  • Author(s): Aoki T, Hikima J
  • Publisher: Eolss Publishers/UNESCO, Co. Ltd., UK
• [Book] Fish And Shellfish Bio-Defense, Section 1. Innate Immunity In Fish. In: Fish Diseases (Ed. Takashi Aoki) (2016)
  • Author(s): Aoki T, Hikima J
  • Publisher: Eolss Publishers/UNESCO, Co. Ltd., UK
• [Book] Diseases Caused By Bacterial Pathogens In Saltwater, Section 5. Vibriosis. In: Fish Diseases (Ed. Takashi Aoki) (2016)
  • Author(s): Takahashi Y, Hikima J
  • Publisher: Eolss Publishers/UNESCO, Co. Ltd., UK