Assessment of inflammatory response by inhibition of complement C5a in zebrafish

• Project/Area Number: 17K15323
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Aquatic life science
• Research Institution: Kyushu University
• Principal Investigator: Nagasawa Takahiro 九州大学, 農学研究院, 助教 (70775444)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 補体 / C5a / アナフィラトキシン / 自然免疫 / 魚類免疫 / ゼブラフィッシュ / 炎症 / 免疫 / 魚類

Outline of Final Research Achievements

We investigated the function of complement C5a in zebrafish, by establishing recombinant zebrafish C5a(recombinant zfC5a; rzfC5a) and anti-zfC5a antibody. The rzfC5a chemo-attracted zebrafish kidney leukocytes in chemotaxis chamber, enhanced phagocytic activity, upregulated expressions of chemokines, suggesting that zfC5a are a key molecule to initiate inflammatory response in early phase of infection. Migration of the leukocytes to zymosan-activated serum was inhibited by anti-zfC5a, suggesting that C5a is a dominant chemoattractant in zebrafish plasma.

Academic Significance and Societal Importance of the Research Achievements

本研究は硬骨魚類の炎症惹起メカニズムを解明し、それら免疫応答を適切に調節することで増養殖の生産性向上を目指すものである。本研究によりC5aを投与または抑制した際の影響が示され、生体内においてC5a産生が免疫活性化、炎症誘引に非常に大きな影響を与えていることがわかった。また作製した組換えzfC5aタンパク及び抗zfC5a抗体はそれらの反応を誘発、抑制するツールとして有用であり、これらを用いてさらなる炎症メカニズム研究の発展が期待できる。

Research Products

• [Journal Article] Generation of virus-specific CD8+ T cells by vaccination with inactivated virus in the intestine of ginbuna crucian carp. (2019)
  • Author(s): Tajimi S, Kondo M, Nakanishi T, Nagasawa T, Nakao M, Somamoto T.
  • Journal Title: Developmental and Comparative Immunology Volume: 93 Pages: 37-44
  • DOI: 10.1016/j.dci.2018.12.009
  • NAID: 120006819494
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Development of Anti-atypical Aeromonas salmonicida Monoclonal Antibodies for Diagnosis of "New Ulcer Disease" in Koi Carp (2018)
  • Author(s): Tomonori Somamoto, Shinpei Maruyama, Takahiro Nagasawa, Miki Nakao, Atsushi Sato, Hajime Hatta, Mitsuru Sato, Yukie Murakami-Yamaguchi, Kumiko Kizu-Mori, Yuki Hirakawa, Hiroshi Narita
  • Journal Title: Fish Pathology Volume: 53 Pages: 36-39
  • NAID: 130006619196
  • Peer Reviewed
• [Journal Article] Development of Anti-atypical <i>Aeromonas salmonicida</i> Monoclonal Antibodies for Diagnosis of &ldquo;New Ulcer Disease&rdquo; in Koi Carp (2018)
  • Author(s): Tomonori Somamoto, Shinpei Maruyama, Takahiro Nagasawa, Miki Nakao, Atsushi Sato, Hajime Hatta, Mitsuru Sato, Yukie Murakami-Yamaguchi, Kumiko Kizu-Mori, Yuki Hirakawa, Hiroshi Narita
  • Journal Title: Fish Pathology Volume: 53 Issue: 1 Pages: 36-39
  • DOI: 10.3147/jsfp.53.36
  • NAID: 130006619196
  • ISSN: 0388-788X, 1881-7335
  • Peer Reviewed
• [Journal Article] Evaluation of probiotic Bacillus aerius B81e isolated from healthy hybrid catfish on growth, disease resistance and innate immunity of Pla-mong Pangasius bocourti. (2017)
  • Author(s): Meidong R, Khotchanalekha K, Doolgindachbaporn S, Nagasawa T, Nakao M, Sakai K, Tongpim S.
  • Journal Title: Fish Shellfish Immunol. Volume: 73 Pages: 1-10
  • DOI: 10.1016/j.fsi.2017.11.032
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Comprehensive validation of T- and B-cell deficiency in rag1-null zebrafish: Implication for the robust innate defense mechanisms of teleosts. (2017)
  • Author(s): Tokunaga Y, Shirouzu M, Sugahara R, Yoshiura Y, Kiryu I, Ototake M, Nagasawa T, Somamoto T, Nakao M.
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 7536-7536
  • DOI: 10.1038/s41598-017-08000-2
  • Peer Reviewed / Open Access
• [Presentation] 飼育下シロワニの飼育環境変化と繁殖周期 (2020)
  • Author(s): 鈴木泰也，牧貴海，中村雅之，長澤貴宏，シロワニ繁殖協議会
  • Organizer: 第６４回水族館技術者研究会
• [Presentation] 魚類免疫学の近年の概況～粘膜免疫と寄生虫対策～ (2020)
  • Author(s): 長澤貴宏
  • Organizer: 第35回九州沖縄ブロック動物園水族館獣医師臨床研究会
  • Invited
• [Presentation] アユのザイモサン結合性レクチンの機能解析 (2020)
  • Author(s): 長澤貴宏・池尻夏菜・ 杣本智軌・中尾実樹
  • Organizer: 令和2年度日本水産学会春季大会
• [Presentation] 軟骨魚類自然抗体の抗原結合能に及ぼす生息地域の影響 (2020)
  • Author(s): 藤村拓実、室谷冬香、野崎玲子、矢澤良輔、長澤貴宏、杣本智軌、中尾実樹、廣野育生、近藤秀裕
  • Organizer: 令和2年度日本水産学会春季大会
• [Presentation] イヌザメ血清レクチンの構造・機能解析 (2020)
  • Author(s): 鳥居爽花・長澤貴宏・中尾実樹
  • Organizer: 令和2年度日本水産学会春季大会
• [Presentation] Production of type-I interferon and the regulatory mechanisms in carp thrombocytes (2019)
  • Author(s): Takahiro Nagasawa, Tomonori Somamoto, and Miki Nakao
  • Organizer: 3rd International Conference on Fish & Shellfish Immunology Gran Canaria
  • Int'l Joint Research
• [Presentation] Direct cytotoxic activity of CD8+ T cells against Ichthyophthirius multifiliis in ginbuna crucian carp, Carassius auratus langsdorfii (2019)
  • Author(s): Masaki Sukeda, Koumei Shiota, Takahiro Nagasawa, Miki Nakao, Tomonori Somamoto
  • Organizer: 3rd International Conference on Fish & Shellfish Immunology Gran Canaria
  • Int'l Joint Research
• [Presentation] 魚類炎症反応におけるC5aの役割解明のための 組換えゼブラフィッシュC5aの作製 (2019)
  • Author(s): 齋藤武尊・長澤貴宏・杣本智軌・中尾実樹
  • Organizer: 令和元年度日本水産学会秋季大会
• [Presentation] Phagocytosis of CD8α+ T cell in ginbuna crucian carp, Carassius auratus langsdorfii (2019)
  • Author(s): Zhao Ge, Takahiro Nagasawa, Miki Nakao, Tomonori Somamoto
  • Organizer: 令和元年度日本水産学会九州支部会
• [Presentation] ドチザメ血清中のザイモサン結合タンパク質の同定 (2019)
  • Author(s): 友井千帆里, 長澤貴宏, 濵 虹花, 杣本智軌, 中尾実樹
  • Organizer: 日本比較免疫学会第31回学術集会
• [Presentation] アフィニティーシルクを用いた魚病細菌除去フィルターの開発 (2019)
  • Author(s): Harsha Prakash、丸山真平、佐藤充、小島桂、佐藤巧視、長澤貴宏、中尾実樹、杣本智軌
  • Organizer: 日本魚病学会
• [Presentation] Functional analysis of two complement C4 isotypes of carp using recombinant proteins (2019)
  • Author(s): Nehlal Rosli, Takahiro Nagasawa, Tomonri Somamoto, Miki Nakao
  • Organizer: The 1st Congress of Asian Society of Developmental and Comparative Immunology
  • Int'l Joint Research
• [Presentation] コイ体表粘液中に存在する補体成分の検出 (2018)
  • Author(s): 吉迫郁子, 黒木将武, 長澤貴宏, 杣本智軌, 中尾実樹
  • Organizer: 日本比較免疫学会学術集会
• [Presentation] 魚類炎症反応におけるC5aの役割解明のための抗ゼブラフィッシュC5a抗体の作成 (2018)
  • Author(s): 斎藤武尊, 長澤 貴宏, 杣本智軌, 中尾実樹
  • Organizer: H31年度日本水産学会春季大会
• [Presentation] 繊毛虫Ichtyophthiriusmultifiliisの成長段階の違いによるギンブナの免疫応答の比較 (2018)
  • Author(s): 塩田昂明, 助田将樹, 中西照幸, 長澤貴宏, 中尾実樹, 杣本智軌
  • Organizer: H31年度日本水産学会春季大会
• [Presentation] 特異抗体を利用したコイ補体成分C5の迅速精製 (2018)
  • Author(s): 中尾実樹, 木村美智代, 長澤貴宏, 杣本智軌
  • Organizer: H31年度日本水産学会春季大会
• [Presentation] VIRAL LIGAND RECOGNITIONS OF CARP THROMBOCYTES AS NATURAL TYPE I INTERFERON PRODUCING CELLS. (2018)
  • Author(s): Takahiro Nagasawa, Tomonori Somamoto, Miki Nakao
  • Organizer: The 14th Congress of the International Society for Developmental and Comparative Immunology
  • Int'l Joint Research
• [Presentation] Type I IFN production by carp thrombocytes as professional antiviral leukocytes, like mammalian plasmacytoid dendritic cells, via IRF7 signaling pathway. (2018)
  • Author(s): Takahiro Nagasawa, Tomonori Somamoto, Miki Nakao
  • Organizer: Asian invertebrate immunity symposium 2018
  • Int'l Joint Research
• [Presentation] PHYLOGENETIC INFERENCE ON FUNCTIONS OF THE CLASSICAL COMPLEMENT PATHWAY IN BONY FISH (2018)
  • Author(s): Miki nakao, Sayo Iwanaga, Takahiro Nagasawa, Tomonori Somamoto
  • Organizer: The 14th Congress of the International Society for Developmental and Comparative Immunology
  • Int'l Joint Research
• [Presentation] 硬骨魚類における古典経路の機能解析から系統発生学的考察 (2018)
  • Author(s): 中尾実樹, 岩永彩代, 長澤貴宏, 杣本智軌
  • Organizer: 第55回日本補体学会学術集会
• [Presentation] ギンブナCD8+Tリンパ球の寄生原虫に対する細胞遊走活性 (2018)
  • Author(s): 助田将樹, 塩田昴明, 長澤貴宏, 中尾実樹, 杣本智軌
  • Organizer: H30年度日本水産学会秋季大会
• [Presentation] Homeostatic functional analysis of Tecrem, a CD46-like complement regulatory protein, on epithelial cells in carp fish (2018)
  • Author(s): Harsha Prakash, Shiori Motobe, Takahiro Nagasawa, Tomonori Somamoto, Miki Nakao
  • Organizer: 平成30年度日本水産学会春季大会
• [Presentation] Ligand recognition specialty of carp thrombocytes as natural type I IFN producing cells. (2017)
  • Author(s): Takahiro Nagasawa, Tomonori Somamoto, Miki Nakao
  • Organizer: The JSFS 85th Anniversary-Commemorative International Symposium「Fisheries Science for Future Generations」
  • Int'l Joint Research
• [Presentation] Functional analysis of tecrem, a CD46-like complement regulatory protein, on epithelial cells in the common carp (2017)
  • Author(s): Harsha Prakash, Shiori Motobe, Takahiro Nagasawa, Tomonori Somamoto, Miki Nakao
  • Organizer: The JSFS 85th Anniversary-Commemorative International Symposium「Fisheries Science for Future Generations」
  • Int'l Joint Research
• [Presentation] コイ栓球によるI型インターフェロン産生機構 (2017)
  • Author(s): 長澤貴宏、杣本智軌、中尾実樹
  • Organizer: 平成29年日本魚病学会秋季大会
• [Presentation] コイ栓球のI型インターフェロン産生能 (2017)
  • Author(s): 長澤貴宏、杣本智軌、中尾実樹
  • Organizer: 平成29年度日本比較免疫学会学術集会
• [Presentation] コイ補体成分B/C2-Bの組換え体作製と機能解析 (2017)
  • Author(s): 赤司小百合、長澤貴宏、杣本智軌、中尾実樹
  • Organizer: 平成29年度日本比較免疫学会学術集会