| 中尾 実樹（なかお みき） | データ更新日：2024.04.18 |
教授 ／
農学研究院
生命機能科学部門
生物機能分子化学講座
| 1. | Trang TT, Nagasawa T, Nakao M, Somamoto T, The expression of CD83 homologs on macrophage subpopulations isolated from brain and kidney in ginbuna crucian carp, 日本水産学会春季大会, 2024.03. |
| 2. | Akhil Kizhakkumpat, Mako I, Prakash H, Nagasawa T, Somamoto T, Nakao M, Development of anti-peptide antibody specific for IgM heavy chain of Oryzias latipes and its application to assay of immune response triggered by BSA-coated microplastics, 日本水産学会春季大会, 2024.03. |
| 3. | 小﨑伶衣、中尾実樹, 魚類およびヒト血清中のメラニン結合タンパク質の検出, 日本水産学会春季大会, 2024.03. |
| 4. | 森 陽理、中尾実樹, システインプロテアーゼによるコイのアレルギー反応について, 日本水産学会春季大会, 2024.03. |
| 5. | Harsha Prakash・Mitsuru Sato・Katsura Kojima・Chisato Sakuma・Akhil Kizhakkumpat・Takahiro Nagasawa・Miki Nakao・Tomonori Somamoto, Preventing parasitic infections caused by Ichthyophthirius multifiliis: A novel approach with a single-chain variable fragment (scFv)-conjugated affinity silk filter device, 日本魚病学会春季大会, 2024.03. |
| 6. | 中尾実樹, CD46様補体制御因子の硬骨魚類における多面的機能, 日本比較免疫学会学術集会, 2023.09. |
| 7. | Zhu J, ＠Nagasawa T, Somamoto T, ＠Nakao M, Molecular cloning of zymosan-binding lectin in the banded houndshark (Triakis scyllium), 日本比較免疫学会学術集会, 2023.09. |
| 8. | 笠 元、長澤貴宏、杣本智軌、中尾実樹, コイ(Cyprinus carpio)栓球はToll様受容体を介して抗原特異的に応答する, 日本比較免疫学会学術集会, 2023.09. |
| 9. | 浅沼里利香、長澤貴宏、中尾実樹、杣本智軌, ギンブナ後腸部に存在する嚢状組織のリンパ組織としての特性, 日本比較免疫学会学術集会, 2023.09. |
| 10. | 椿野 鈴, 吉井 啓亮, 岡本 裕之, 大野 薫, 牧野 由美子, 長澤 貴宏, 杣本 智軌, 中尾 実樹, エドワジエラ症耐性ヒラメの粘膜免疫因子の探索, 日本比較免疫学会学術集会, 2023.09. |
| 11. | Tran Thu Trang, Takahiro Nagasawa, Miki Nakao, Tomonori Somamoto, Expression of CD83 isotypes during the differentiation of monocyte-macrophage lineage in ginbuna crucian carp, 日本比較免疫学会学術集会, 2023.09. |
| 12. | 金田誠正、長澤貴宏、杣本智軌、中尾実樹, コイ血清中の溶血反応阻害タンパク質の精製と同定, 日本補体学会学術集会, 2023.08. |
| 13. | 金田誠正, 長澤貴宏, 杣本智軌, 中尾実樹, コイの可溶性補体制御因子の精製, 日本魚病学会春季大会, 2023.03. |
| 14. | Miki Nakao, Mucosal innate defense in carp fish involving T cells, complement system, and epithelial cells, The 2nd Asian Society of Developmental and Comparative Immunology Congress, 2022.10, Mucosal immunity is regarded as a crucial first-line defense against microbial infection in teleost fish, as supported by identification of well-developed mucosa-associated lymphoid organs, secretary forms of IgM and IgT, mucus lectins and antimicrobial peptides. Innate immune effector mechanisms, however, remain to be elucidated by functional evidence. We have identified CD8+ T cells of ginbuna crucian carp as effector cells to kill extracellular parasites, Ichthyophthirius multifiliis, in a contact-dependent and MHC-independent manner via serine proteases and perforin, suggesting the ancestral innate function of teleost T cells. On the other hand, presence of components of the complement system, a major humoral innate immune factor, in teleost skin has been reported with less information on their local activation and effector functions. Recently we have detected activation of the classical complement pathway towards Aeromonas hydrophila and A. salmonicida in carp skin mucus. This activation proceeds at least until the reaction step of C3, suggesting that the mucus complement is functional as an opsonic system. Interestingly, epithelial cells express a CD46-like regulator of complement activation (Tecrem) and stimulation of Tecrem seems to enhance robustness of epithelial sheet by up-regulating tight junction proteins in carp and ginbuna crucian carp. Since C3b, an activated form of C3, is a major ligand of Tecrem, mucosal complement activation may also contribute to strengthen the epithelial barrier against microbial infection.. |
| 15. | 畠山紗矢香・長澤貴宏・杣本智軌・中尾実樹, コイ血清中におけるC3の二量体形成, 日本補体学会学術集会, 2022.09, [URL]. |
| 16. | 又吉百音, 長澤貴宏, 杣本智軌, 中尾実樹, 細胞内補体活性化の系統発生学的検討:コイ末梢血白血球における補体C3の断片化, 日本比較免疫学会学術集会, 2022.08. |
| 17. | 又吉百音・長澤貴宏・杣本智軌・中尾実樹, 魚類におけるC3の細胞内活性化の可能性, 日本補体学会学術集会, 2022.08, [URL]. |
| 18. | 白山智恵、＠長澤貴宏、中尾実樹, ドチザメ血清中の抗菌因子の探索と同定, 令和4年度 日本水産学会春季大会, 2022.03, 【背景・目的】 微生物による感染防御に重要な免疫系には、獲得免疫と自然免疫の2つの機構がある。脊椎動物における両免疫機構の進化を深く理解するためには、より原始的な動物の免疫系を調査することが重要である。本研究の目的は、原始的な脊椎動物モデルとして軟骨魚類、ドチザメ（Triakis scyllium）を用い、その血清中の微生物結合因子および抗菌因子の検出・同定を試みた。 【材料・方法】ドチザメ血清300 µLをSuperdex 200 increaseカラム（1 x 30 cm）を使用したゲル濾過で分画し、各フラクション（Fr.）の1) Micrococcus luteusに対する溶菌活性、2) M. luteus, Aeromonas hydrophilla, Staphylococcus aureus, Edwardsiella tarda, Streptococcus parauberis, およびFlavobacterium sp.に対する凝集活性を測定した。また、各フラクションに含まれるタンパク質をSDS-PAGEで分析した。 【結果・考察】 Superdex 200 increaseの限界排除分子量に相当するFr. 7〜8に、S. parauberis を除く全菌種に対する凝集活性が検出された。また、E. tardaおよびS. aureusに対しては、分子量数十万〜数万に相当する広い範囲のFr.が凝集活性を示した。一方、分子量約1〜２万に相当するFr. 24〜27に、M. luteusに対する溶菌活性が認められた。各フラクションのSDS-PAGEによって、Fr. 7〜8には、IgM抗体のH鎖と考えられる分子量約75,000のバンドが認められた。また、Fr. 24〜27は、分子量14,000の1本のバンドを示した。このポリペプチドのN末端アミノ酸配列は、YVYSKXELARTLRRNGLDGYと解読され、これはジンベイザメのLysozyme C-1-likeと有意な相同性（80%）を示した。したがって、分子量約14,000の溶菌因子は、ドチザメのリゾチームと同定された。 . |
| 19. | 吉迫郁子・長澤貴宏・杣本智軌・中尾実樹, コイ体表粘液中の補体成分と補体活性化, 日本補体学会学術集会, 2021.09, [URL]. |
| 20. | 阪中晴子・長澤貴宏、中尾実樹、杣本智軌, 粘膜組織を介した抗原投与法の違いによるギンブナ抗体産生応答の比較, 日本魚病学会, 2021.09. |
| 21. | Nguyen Quang Khai, Harsha Prakash, Takahiro Nagasawa, Miki Nakao, Tomonori Somamoto, The interaction between Koi Herpesvirus and a CD46-like regulator of complement activation in teleost. , 日本魚病学会, 2021.09. |
| 22. | Akhil K, Nawasawa T, Somamoto T, Oshima Y, Nakao M., Adsorption of protein and bacteria on microplastics: Effect on oral intake and adaptive immunity of Oryzias latipes, 日本水産学会九州支部総会, 2021.09. |
| 23. | Prakash H, ＃Sukeda M, Nawasawa T, Nakao M, Somamoto T, Characterization of cell surface receptor NCCRP-1 on CD8+T cells of Ginbuna crucian carp in the cytotoxic activity against Ichthyophthirius multifiliis, 日本水産学会九州支部総会, 2021.09. |
| 24. | 笠 元・長澤貴宏・ 杣本智軌・中尾実樹, コイ(Cyprinus carpio)栓球による免疫応答のパターン認識機構, 日本水産学会九州支部総会, 2021.09. |
| 25. | 朱 珈凝, 友井千帆里, 長澤貴宏, 杣本智軌, 中尾実樹, ドチザメ血清中のザイモサン結合レクチンの同定と発現解析, 日本比較免疫学会学術集会, 2021.08. |
| 26. | 吉迫郁子, 長澤貴宏, 杣本智軌, 中尾実樹, コイ体表粘液中に存在する補体成分と粘液における補体活性化, 日本比較免疫学会学術集会, 2021.08. |
| 27. | 又吉百音・長澤貴宏・杣本智軌・中尾実樹, 魚類における補体成分C3の細胞内活性化, 日本水産学会九州支部総会, 2021.01. |
| 28. | 鳥居爽花・長澤貴宏・中尾実樹, イヌザメ血清レクチンの構造・機能解析, 日本水産学会春季大会, 2020.03. |
| 29. | Rosli N, Yamamoto A, Nagasawa T, Somamoto T, Nakao M, Functional analysis of two complement C4 isotypes of carp using recombinant proteins, The 1st Congress of Asican Society of Developmental and Comparative Immunology, 2019.11. |
| 30. | 友井千帆里, 長澤貴宏, 濵 虹花, 杣本智軌, 中尾実樹, ドチザメ血清中のザイモサン結合タンパク質の同定, 日本比較免疫学会学術集会, 2019.09, [URL]. |
| 31. | 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, 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, 日本魚病学会, 2019.09. |
| 32. | 齋藤武尊・長澤貴宏・杣本智軌・中尾実樹, 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, 日本水産学会秋季大会, 2019.09. |
| 33. | 中尾実樹, 魚類補体研究の最近の進展と動向, 日本補体学会学術集会, 2019.08, [URL]. |
| 34. | Masaki Sukeda, Koumei Shiota, Takahiro Nagasawa, Miki Nakao, Tomonori Somamoto, Direct cytotoxic activity of CD8+ T cells against Ichthyophthirius multifiliis in ginbuna crucian carp, Carrassius auratus langsforfii, 3rd International Conference of Fish and Shellfish Immunology, 2019.06, [URL], A line of studies has shown that several humoral immune factors includingcomplement, lectins and antibodies are involved in protection fromparasite infections. However, cell-mediated immunity against parasiteshas poorly been understood in teleostfish. In the present study, directcytotoxic activity of leukocytes againstIchthyophthirius multifiliishas beendemonstrated in ginbuna crucian carp. Leukocytes labeled by eachmonoclonal antibody (2C3: anti-CD8, 6D1:anti-CD4, GB20: anti-macro-phages/neutrophils) were co-incubated withI. multifiliis. Thefluorescentmicroscopic observation showed that CD8+T cells from naïve ginbunacarp, but not other leucocytes, contactedI. multifilii. The cytotoxic activityof CD8+T cells was significantly higher than that of other leucocytes,indicating that CD8+T cells are dominant effector cells againstI. multifiliis.The cytotoxic assay using a trans-well insert suggested that CD8+T cellsrequire to contact the parasites for the direct killing. Furthermore, a serineprotease inhibitor 3, 4-dichloroisocoumarin (DCI) inhibited the cytotoxicactivity of CD8+Tcells, but a perforin inhibitor Concanamycin A (CMA) didnot. These results indicate that teleost CD8betaT cells have natural cell-mediated cytotoxicity against extracellular parasite by utilizing serineproteases, such as granzyme, suggesting that CD8+T cells play animportant role in innate immunity against extracellular protozoanparasites.. |
| 35. | Takahiro Nagasawa, Tomonori Somamoto, Miki Nakao, TLR-mediated type-I interferon production and the regulatory mechanisms in carp thrombocytes, 3rd International Conference of Fish and Shellfish Immunology, 2019.06, [URL], In early stage of viral infection, type-I interferons (IFNs) are produced by signaling from innate immune receptors such as Toll-like receptors (TLRs), which recognize virus-specific molecular patterns including nucleic acids. The type-I IFN transcriptions induced by TLRs are regulated via nuclear factor kB (NF-kB) complex and interferon regulatory factors (IRFs), but these details are still unclear in fish. In the present study, we show that thrombocytes in common carp (Cyprinus carpio) have a potent ability to produce large amount of IFNs in response to TLR signaling. Magnetic- sorted HB-8 mAbþ carp thrombocytes and negatively sorted other pe- ripheral blood leukocytes (PBLs) were incubated with resiquimod (also called R848, a potent agonist of TLR7/8), followed by qPCR analysis. The expression levels of the common carp type-I IFNs (ccIFN1 and ccIFN2) in thrombocytes were considerably higher compared with that of in other PBLs. Whereas the ccIFN1 expression was relatively lower than the ccIFN2, the R848 stimulant highly upregulated the ccIFN1 expression than ccIFN2. Although typical inflammatory cytokines including interleukin-6 were also upregulated in thrombocytes, the expression levels were still lower than those in other PBLs. These results indicate that activation of carp thrombocytes by R848 inclines immune system toward antiviral response, rather than inflammation. Expression levels of IRF3 and IRF7 were also upregulated by R848, implying that the IFN transcriptions were activated by these IRFs. The expression of the IFNs and inflammatory cytokines were decreased by several NF-kB signaling inhibitors such as BAY11-7082 or phenethyl caffeate, however, sensitivities to each inhibitor were different between the IFNs and other cytokines. In the presence of those inhibitors, the ccIFN2 expression was correlated with the level of IRF3. In contrast, ccIFN1 expressions seem to be linked to IRF7, suggesting that these two IRFs regulates different IFN genes separately. Our finding suggests that fish thrombocytes are important components for antiviral immunity and can be a new target for the strategy of disease control and vaccine development. . |
| 36. | 中尾実樹, 木村美智代, 長澤 貴宏, 杣本智軌, 特異抗体を利用したコイ補体成分C5の迅速精製, 日本水産学会春季大会, 2019.03, 【目的】魚類における補体の自然免疫機能を正確に理解するには、タンパク質レベルでの機能解析が必要である。しかし、補体成分の多くは、魚類では特に不安定で、活性を保持した補体成分の精製が困難である。本研究では、特異抗体のアフィニティーカラムを用い、活性を保持した補体成分C5を高純度に精製するための、迅速法の確立を目指した。 【方法】(1)抗コイC5の作成：コイC5-1アイソタイプのnetrinドメインを、N末端に6xHisタグを付加した組換タンパク質として大腸菌で発現させ、Ni-カラムで精製後、ウサギに免疫注射して抗血清を得た。 (2)アフィニティーカラムの調製： L-lysineおよび抗コイC5-1ウサギIgGをNHS-活性化HiTrapカラム（各1 ml）にそれぞれ固定化後、TBSで平衡化した。(3)溶血活性測定：抗体感作ヒツジ赤血球を標的としたCH50補体価測定法に準拠した。 【結果】コイ血清（10 ml）を、直列に接続したLysine-HiTrap→抗C5-HiTrapカラムにシリンジで負荷した。この直列カラムを20 mlのTBSで洗浄後、Lysine-HiTrapカラムを外し、抗C5-HiTrapカラム出口をHiTrap Desaltingカラム入口に接続した。5 mlの0.1 M glycine-HCl (pH 2.5)を送って抗C5-HiTrapカラムにトラップされたC5を脱離させると同時に、TBSへの緩衝液交換を済ませた。本法により1.5時間以内で精製されたC5は、SDS-PAGEでα鎖(110 kDa)とβ鎖(70 kDa)の典型的な二本鎖構造を示し、C5除去コイ血清と組み合わせると溶血活性を保持していることが確認された。 . |
| 37. | 斎藤武尊, 長澤 貴宏, 杣本智軌, 中尾実樹, , 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, 日本水産学会春季大会, 2019.03. |
| 38. | 塩田昂明, 助田将樹, 中西照幸, 長澤 貴宏, 杣本智軌, 中尾実樹,, 繊毛虫Ichtyophthirius multifiliisの成長段階の違いによるギンブナの免疫応答の比較, 日本水産学会春季大会, 2019.03. |
| 39. | Nakao M, Iwanaga S, Nagasawa T, Somamoto T, Evolutionary implication on the function of the classical pathway of the complement system, Asian Invertebrate Immunity Symposium, 2018.09. |
| 40. | Nagasawa T, Somamoto T, Nakao M, Type I IFN production by carp thrombocytes as professional antiviral leukocytes, like mammalian plasmacytoid dendritic cells, via IRF signaling pathway, Asian Invertebrate Immunity Symposium, 2018.09. |
| 41. | Rosli N, Yamamoto A, Nagasawa T, Somamoto T, Nakao M, Functional analysis of two complement C4 isotypes of carp using recombinant proteins, 日本水産学会秋季大会, 2018.09. |
| 42. | 助田将樹,塩田昂明,長沢貴宏,中尾実樹,杣本智軌, 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, 日本水産学会秋季大会, 2018.09. |
| 43. | 中尾実樹, 岩永彩代、長沢貴宏、杣本智軌、, 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, 日本補体学会学術集会, 2018.08. |
| 44. | 吉迫郁子, 黒木将武, 長澤貴宏, 杣本智軌, 中尾実樹, コイ体表粘液中に存在する補体成分の検出, 日本比較免疫学会学術集会, 2018.08. |
| 45. | Nakao M, Iwanaga S, Nagasawa T, Somamoto T, Phylogenetic inference on functions of the classical complement pathway in bony fish, 14th Congress of International Society of Developmental and Comparative Immunology, 2018.06, [URL], Bony fish is one of the ancestral vertebrates that possess immunoglobulins, on which the classical pathway of complement system relies for target recognition. To access the role of the classical pathway in bony fish complement, we have analyzed the hemolytic reaction and C3b-deposition on the target surface of the common carp (Cyprinus carpio) complement, using carp serum immunochemically depleted of factor D (Df), a serine protease responsible for activation of the alternative pathway, an activation and amplification cascade that bypasses the classical pathway. To our surprise, the Df-depletion abolished hemolytic activities of the serum both through the classical and alternative pathways, when assayed using antibody-sensitized and non-sensitized sheep and rabbit erythrocytes. However, a low level of C3b-deposition, probably resulted from the classical pathway activation in the absence of Df was demonstrated on various target surfaces by flow cytometry and ELISA. In addition, Df-depletion inhibited C5-deposition essential for the cytotoxic complex formation on the target cells. The C3b-bound erythrocytes were hemolyzed by serum but easily lysed by Mg-EGTA-serum, probably triggering the alternative pathway amplification more efficiently than unbound erythrocytes. These results, taken together, suggest that an ancestral classical pathway alone lacks C5-activating ability but tags target cells with a small amount of C3b, which triggers enhanced C3-activation by the alternative pathway, leading to completion of the terminal cytolytic pathway.. |
| 46. | Nagasawa T, Somamoto T, Nakao M, Viral ligands recognition of carp thrombocytes as natural type I interferon producing cells, 14th Congress of International Society of Developmental and Comparative Immunology, 2018.06, [URL]. |
| 47. | 助田将樹, 長澤 貴宏, 中尾 実樹, 杣本智軌, ギンブナCD8+Tリンパ球の細胞外寄生原虫に対する細胞障害機構の解明, 日本水産学会春季大会, 2018.03. |
| 48. | Prakash H, Motobe S, Nagasawa T, Somamoto T, Nakao M, Homeostatic functional analysis of Tecrem, a CD46-like complement regulatory protein, on epithelial cells in carp fish, 日本水産学会春季大会, 2018.03. |
| 49. | Nakao M, Noguchi M, Akahoshi S, Nagasawa T, Somamoto T, Functional diversity of two C7 isotypes in bony fish, a primitive vertebrate model, European Meeting on Complement in Human Disease, 2017.09, [URL]. |
| 50. | Prakash H, Motobe S, Nagasawa T, Somamoto T, Nakao M, Functional analysis of Tecrem, a CD46-like complement regulatory protein, on epithelial cells in the common carp, The JSFS 85th Anniversary-Commemorative International Symposium, 2017.09. |
| 51. | 丸山真平、佐藤充、小島桂、佐藤巧視、長沢貴宏、中尾実樹、杣本智軌, アフィニティーシルクの魚病細菌感染症診断への応用, 日本魚病学会秋季大会, 2017.09. |
| 52. | 助田将樹、長沢貴宏、中尾実樹、杣本智軌, 繊毛虫Icthyophthirius multifiliisに対する魚類CD8+T細胞の傷害機構, 日本魚病学会秋季大会, 2017.09. |
| 53. | 赤司小百合、長沢貴宏、杣本智軌、中尾実樹, コイ補体成分B/C2-Bの組換え体作成と機能解析, 日本比較免疫学会学術集会, 2017.08. |
| 54. | 長沢貴宏、杣本智軌、中尾実樹, コイ栓球のI型インターフェロン産生能, 日本比較免疫学会学術集会, 2017.08. |
| 55. | 黒木将武、吉岡和紀、長沢貴宏、杣本智軌、中尾実樹, コイ補体Properdinアイソタイプの機能解析, 日本補体学会学術集会, 2017.08. |
| 56. | 野口真代、長沢貴宏、杣本智軌、中尾実樹, コイ補体C7アイソタイプの機能解析, 日本補体学会学術集会, 2017.08. |
| 57. | 新内悠介, 満崎敬子, 福田圭佑, 長澤 貴宏, 中尾 実樹, 杣本智軌, ギンブナの細胞性免疫抑制によるKHV病発症, 日本水産学会春季大会, 2017.03. |
| 58. | 助田将樹, 長澤 貴宏, 中尾 実樹, 杣本智軌, 繊毛虫 Ichthyophthirius multiliis に対するギンブナT 細胞の傷害活性, 日本水産学会春季大会, 2017.03. |
| 59. | 野口真代, 長澤 貴宏, 中尾 実樹, 杣本智軌, コイ補体 C7 アイソタイプの機能解析, 日本水産学会春季大会, 2017.03. |
| 60. | Harsha Prakash, Shiori Motobe, Takahiro Nagasawa, Tomonori Somamoto, Miki Nakao, Expression and homeostatic functions of Tecrem, a CD46-like complement regulatory protein on epithelial cells in bony fish., 26th International Complement Workshop, 2016.09, [URL], Mammalian CD46 has been reported as a multitasking immune modulator, which regulates complement activation on host cells, T cell-mediated adaptive responses, and wound repair by involving epithelial cells. It is of particular interest to explore the evolutionary path of such versatile functions of CD46. Our group has identified a CD46-like molecule, termed teleost complement regulatory membrane protein or Tecrem, in a few cyprinid fish species and has shown its regulatory function on complement activation at the protein level. Furthermore, we have also found that Tecrem expressed on T cells modulates mitogen-induced T cell proliferation in ginbuna crucian carp, indicating that modulation of adaptive immunity is one of the evolutionarily conserved functions of the CD46-like complement regulator. In the present study, we have explored a homeostatic role of Tecrem in the maintenance of fish epithelium, by analyzing expression behavior of Tecrem on an epithelial cell line (KF-1) derived from carp fin. Flow cytometric analysis of Tecrem expression on KF-1 using anti-carp Tecrem monoclonal antibody (MAb) (1F12) suggested that Tecrem expression may be affected by cell aggregation and adhesion. Fluorescent microscopic observation and ELISA-based assay for Tecrem also indicated a role of Tecrem in the adhesion of KF-1 to the surface of culture media. Furthermore, 1F12 MAb deposited on the culture plate significantly enhanced an early stage of cell adhesion process of KF-1. Towards further functional analyses of carp Tecrem, we have prepared recombinant Tecrem proteins in the bacterial expression system. Among the four short consensus repeat (SCR) modules making up the extracellular domains of Tecrem, the N-terminal two SCRs (rSCR1-2) and the C-terminal two SCRs (rSCR3-4) were separately expressed as 6xHis-tagged soluble protein using pCold-I vector and Origami B strain. Interaction of the two recombinant domains with carp C3 isotypes and their inhibition of carp complement activation cascades are currently analyzed in parallel with raising polyclonal antibodies, for the clarification of functional modules and their mode of action. . |
| 61. | Kazuki Yoshioka, Yoko Kato-Unoki, Takahiro Nagasawa, Tomonori Somamoto, Miki Nakao, Carp properdin: structural and functional diversity of two isotypes., 26th International Complement Workshop, 2016.09, [URL], The alternative pathway (ACP) of the complement system is an antibody-independent activation pathway, in which properdin (Pf) has been known as a positive regulator of the activation and a possible pattern-recognition molecule to trigger ACP activation. Teleost complement system has a striking feature that some of its components are diversified into multiple isoforms with different functions. However this diversity is less characterized for teleost Pf, especially at the protein level. The present study was aimed at elucidating isotypic diversity and functional differentiation of Pf in the common carp. Molecular cloning of carp Pf revealed two distinct full-length cDNA sequences, CaPf1 and CaPf2, that predicts mature proteins composed of seven thrombospondin type1 domains (TSP0-TSP6), sharing 77% amino acid sequence identity. Genomic Southern hybridization suggested that CaPf1 and CaPf2 are encoded by single each gene in carp genome. Real-time quantitative PCR indicated that expression level of CaPf1 is most abundant in the spleen, whereas CaPf2 was detected mainly in head kidney and renal kidney. Rabbit antibodies were raised against their recombinant proteins corresponding to TSP4-6 domains. Western blotting using anti-CaPf1 and anti-CaPf2 revealed that CaPf1 and CaPf2 are mainly present as a hexamer of polypeptide with molecular weights of 49,000 and 48,000, respectively, in carp serum. Interestingly, CaPf1 and CaPf2 showed different spectra of binding to various microbes, suggesting their functional diversity.. |
| 62. | 木村美智代, 畑中大作, 一木昭土, 長澤 貴宏, 杣本 智軌, 和合治久, 中尾 実樹, コイ血清レクチン（MFAP4）の構造および機能解析, 日本比較免疫学会学術集会, 2016.08. |
| 63. | 鵜木（加藤）陽子, 杣本 智軌, 中尾 実樹, 獲得免疫欠損ゼブラフィッシュにおけるミエロペルオキシダーゼ遺伝子のノックアウト, 日本水産学会春季大会, 2016.03. |
| 64. | 岡田幸浩, 杣本 智軌, 中尾 実樹, 長澤 貴宏, 畑中晃昌, 青木直人, 平澤徳高, 梅田奈央子, 大豆タンパク質含有飼料が魚類の抗体産生応答に与える影響, 日本水産学会春季大会, 2016.03. |
| 65. | Kazuki Yoshioka, Yoko Kato-Unoki, Tomonori Somamoto, Miki Nakao, Structural and functional diversity of properdin isotypes in the common carp complement system. , 13th Congress of the International Society for Developmental and Comparative Immunology, 2015.06, [URL], The alternative pathway (ACP) of the complement system is an antibody-independent activation pathway, in which properdin (Pf) has been known as an essential positive regulator of the activation and possibly as a pattern-recognition molecule to trigger ACP activation. Teleost complement system has a striking feature that some of its components are diversified into multiple isoforms with different functions. However this diversity is less characterized for teleost Pf, especially at the protein level. The present study was aimed at elucidating isotypic diversity and functional differentiation of Pf in the common carp. Molecular cloning of carp Pf revealed two distinct full-length cDNA sequences, CaPf1 and CaPf2, that predicts mature proteins composed of seven thrombospondin type1 domains (TSP0-TSP6), sharing 77% amino acid sequence identity. Genomic Southern hybridization suggested that CaPf1 and CaPf2 are encoded by single each gene in carp genome. Real-time quantitative PCR indicated that expression level of CaPf1 is most abundant in the spleen, whereas CaPf2 was detected mainly in head kidney and renal kidney. Rabbit antibodies were raised against their recombinant proteins corresponding to TSP4-6 domains. Western blotting using anti-CaPf1 and anti-CaPf2 revealed that CaPf1 and CaPf2 are mainly present as a hexamer of polypeptide with molecular weights of 49,000 and 48,000, respectively, in carp serum. Interestingly, CaPf1 and CaPf2 showed different spectra of binding to various microbes, suggesting their functional diversity.. |
| 66. | 満崎敬子, 中尾 実樹, 杣本 智軌, コイとギンブナにおけるコイヘルペスウイルスへの免疫応答の比較, 日本水産学会春季大会, 2015.03. |
| 67. | 吉岡和紀, 鵜木陽子, 杣本 智軌, 中尾 実樹, コイ補体 Properdin アイソフォームの機能解析, 日本水産学会春季大会, 2015.03. |
| 68. | 前田佑佳, 杣本 智軌, 鶴田幸成, 中尾 実樹, 穴あき病発症時におけるコイ補体成分の組織内分布, 日本水産学会春季大会, 2015.03. |
| 69. | 赤木良太, 長澤貴宏, 杣本 智軌, 中尾 実樹, コイB細胞における自然免疫活性化能, 日本水産学会九州支部大会, 2015.01. |
| 70. | 菅原亮太, 吉浦康寿, 杣本 智軌, 中尾 実樹, Rag1欠損ゼブラフィッシュの免疫調節機構, 日本水産学会秋季大会, 2014.09. |
| 71. | 元部詩織, 辻倉正和, 中村亮太, 杣本 智軌, 中尾 実樹, CD46 様コイ補体制御因子（Tecrem）の上皮細胞における表面発現動態, 第51回補体シンポジウム, 2014.08. |
| 72. | 吉岡和紀, 鵜木陽子, 杣本 智軌, 中尾 実樹, コイ補体 Properdin アイソフォームの多様性と機能解析, 日本比較免疫学会学術集会, 2014.07. |
| 73. | 長澤貴宏, 中易千早, Aja M. Rieger, Daniel R. Barreda, 杣本 智軌, 中尾 実樹, 魚類栓球の異物分解・殺菌作用, 日本水産学会春季大会, 2014.03. |
| 74. | 柳 綾佳, 畑中晃昌, 青木直人, 杣本 智軌, 中尾 実樹, 無魚粉飼料が魚類の補体に及ぼす影響, 日本水産学会秋季大会, 2013.09. |
| 75. | 多治見誠亮, 杣本 智軌, 中西照幸, 中尾 実樹, 不活化ウイルスの経腸管感作によって誘導されるギンブナT細胞の免疫応答及び感染防御効果, 日本水産学会秋季大会, 2013.09. |
| 76. | Indriyani Nur, 原田光利, 杣本 智軌, 中尾 実樹, 中村亮太, 辻倉正和, Characterizations of membrane-bound complement regulatory protein in Ginbuna Crucian Carp Carassius auratus langsdorfii, 日本比較免疫学会学術集会, 2013.08. |
| 77. | 長澤貴宏, 中易千早, 杣本 智軌, 中尾 実樹, コイ栓球は他の白血球により活性化され貪食能を示す, 日本水産学会春季大会, 2013.03. |
| 78. | 豊福太樹, 山下基子, 長澤貴宏, 杣本 智軌, 中尾 実樹, ギンブナ樹状細胞マーカー CD83 に対する抗体の作製と発現, 日本水産学会春季大会, 2013.03. |
| 79. | 白水正道, 杣本 智軌, 吉浦康寿, 中尾 実樹, Rag1欠損ゼブラフィッシュにおけるT細胞の機能不全の検証, 日本水産学会九州支部大会, 2013.01. |
| 80. | 福田圭佑, 杣本 智軌, 中尾 実樹, ギンブナにおけるコイヘルペスウイルスに対する防御機構の解明, 日本水産学会九州支部大会, 2013.01. |
| 81. | 中村亮太, 辻倉正和, 杣本 智軌, 中尾 実樹, コイ膜型補体制御因子cTecremのモノクローナル抗体作製と機能解析, 日本水産学会九州支部大会, 2013.01. |
| 82. | 鵜木陽子, 辻倉正和, 一木智子, 杣本 智軌, 中尾 実樹, コイ補体成分ProperdinのcDNAクローニングと構造解析, 日本水産学会九州支部大会, 2013.01. |
| 83. | 中村亮太、原田光利、長澤貴宏、Indriyani Nur、辻倉正和、一木智子、杣本智軌、中尾実樹, 魚類CD46様補体制御因子の多様性と機能の解析, 補体シンポジウム, 2012.08. |
| 84. | Takahiro Nagasawa, Chihaya Nakayasu, Tomomasa Matsuyama, Aja M. Rieger, Daniel R. Barreda, Tomonori Somamoto, Miki Nakao, Phagocytosis and bactericidal abilities of teleost thrombocytes, 12th Congress of International Society of Developmental and Comparative Immunology, 2012.07, Thrombocytes have been recognized as haemostatic cells in non-mammalian vertebrates. Unlike mammalian platelets, thrombocytes are nucleated cells with a lymphocyte-like morphological feature, and possible involvement of thrombocytes in innate immune function has been considered in addition to the haemostatic function. In the present study, we report phagocytic abilities of some teleost thrombocytes. Using a monoclonal antibody specific for thrombocytes of common carp (Cyprinus carpio), thrombocytes were isolated from peripheral blood and examined for expression of various immune-related genes, resulting in detection of significant level of lysozyme, iNOS and MHC class II using RT-PCR. Upon flow cytometry-based phagocytosis assay, a number of thrombocytes ingested fluorescent latex particles (0.5 μm, 1 μm, 2 μm and 3 μm), bacteria (Escherichia coli) and zymosan particles. Phagocytosis by the thrombocytes was also confirmed by fluorescent microscopy and transmission electron microscopy, which revealed internalization of these particles into thrombocytes. We also observed that thrombocytes of the olive flounder (Paralichthys olivaceus) had similar phagocytic behavior. These data indicate that thrombocytes of those species are potent phagocytes, suggesting that those phagocytic characteristics of thrombocytes are widely conserved in teleosts. We also assessed a phagolysosome formation ability of teleost thrombocytes against the ingested pathogens, for detecting intracellular bactericidal activities of those thrombocytes. By using an ImageStream multi-spectral flow cytometer, we assessed phagolysosome fusion of goldfish (Carassius auratus) thrombocytes. On this analysis we detected that lysosomes of these thrombocytes visualized with fluorescent dextran were co-localized with the ingested small beads and formed a ring around large beads like other typical phagocytes. Overall, the results indicate that teleost thrombocytes have dual functions as not only haemostatic cells but also as phagocytic immune cells against microbial infections.. |
| 85. | Haruka Tsukamoto, Yutaka Fukuda, Tomonori Somamoto, Miki Nakao, Functions of CD8-positive and CD4-positive lymphocytes against virus-infection in ginbuna crucain carp, 12th Congress of International Society of Developmental and Comparative Immunology, 2012.07, Two serotypes of Streptococcus parauberis, pathogens of streptococcosis in Japanese flounder (Paralichthys olivaceus), have caused severe losses of the livestock, due to the lack of effective vaccine and its resistance to antibiotics approved for flounder culture. These strains show different virulence, but their mechanisms of their pathogenicity are totally unknown, and identification of virulence factors are needed for effective vaccine development. The present study, therefore, was aimed at clarifying effect of S. parauberis culture supernatant on flounder. Since the diseased flounder shows anaemia, we examined the effects of S. parauberis on flounder peripheral blood. As a result, S. parauberis -injected flounder showed decreased numbers of peripheral erythrocytes, suggesting that S. parauberis may produce a haemolytic factor. To characterize the haemolytic factor, a supernatant of S. parauberis culture (25°C for 48h in Todd Hewitt broth) was mixed with sheep erythrocytes, resulting in significant level of haemolysis. The haemolytic factor was stable on heating at 100°C for 10 min, and passed through an ultrafiltration membrane with a 5 kDa cut off limit (Amicon Ultra15), indicating that the factor is not proteinaceous haemolysin but a heat-resistant low molecular mass substance. We also investigated the effects of the culture supernatant on flounder leukocytes. Peripheral leukocytes were separated from flounder and carp using Percoll discontinuous density gradient centrifugation and cultured with the supernatant in 96-well plate. The stimulated flounder cells showed agglutination and significant proliferation as assayed by BrdU uptake, while carp cells did not. These results suggest that S. parauberis secrets a mitogen specific for flounder leucocytes. Characterization of the mitogen and resulting leucocyte response is in progress.. |
| 86. | Tomokazu Yamaguchi, Kazufumi Takamune, Masakazu Kondo, Yukinori Takahashi, Miki Nakao, Yoko Kato-Unoki, Tamotsu Fujii, Identification of Pattern Recognition Molecules in Hagfish Complement System, 12th Congress of International Society of Developmental and Comparative Immunology, 2012.07, All extant jawed vertebrates share a common adaptive immune system in which immunoglobulin domain-based molecules act as antigen receptors. On the other hand, jawless vertebrates, lamprey and hagfish, use variable lymphocyte receptors composed of leucine-rich repeat cassettes for antigen recognition. Since invertebrates and primitive chordates do not have an adaptive immune system, the immune system seems to change dramatically in the course of evolution from primitive chordates to gnathostomes. From a phylogenic perspective of defence mechanisms, previously we found complement C3 and mannose-binding lectin-associated serine protease 1 (MASP-1) in hagfish and suggested the involvement of lectin pathway in hagfish innate immune system. In this study, we focused on the pattern recognition molecules in the hagfish, Eptatretus burgeri, and tried to purify them from serum by affinity chromatography. When the serum was treated with GlcNAc-agarose, followed by successive elution of the binding molecules with GlcNAc and EDTA, mainly four proteins (31 kDa, 27 kDa, 26 kDa, and 19 kDa) and 26 kDa protein were detected in the GlcNAc-eluate and EDTA-eluate, respectively. Collagenase treatment showed the presence of collagen-like domain only in the 26 kDa protein in the EDTAeluate. Since common pattern recognition molecules such as mannose-binding lectins possess collagen-like domains, we examined the entity of the 26 kDa protein by sequencing its N-terminal amino acids and cDNA obtained by 3’ and 5’ RACE methods, and identified it as a member of C1q family. Herein, it will be referred to as hagfish C1q (hagC1q). Western blot analyses using anti-hagC1q, MASP-1, and complement C3 antibodies showed that hagC1q associated with MASP-1 and complement C3 in the serum and had binding ability to Escherichia coli as a divalent cation-dependent manner. These results suggest that hagC1q plays an important role in hagfish innate immune system.. |
| 87. | Vo Kha Tam, Chie Okura, Masakazu Kondo, Tomonori Somamoto, Miki Nakao, Isotypic diversity in the ontogenetic expression of the complement component in the common carp (Cyprinus carpio), 12th Congress of International Society of Developmental and Comparative Immunology, 2012.07, Isotypic diversity of complement components is a striking feature of the teleost complement system. As a first line of innate defence, the complement system has been considered as an important clue to humoral defence in early development however functional diversity of the isotypes during teleost ontogeny is poorly understood. The present study aimed at clarifying comprehensive picture of ontogenetic expression of the diversified complement component isotypes in carp. Real-time quantitative PCR detected embryonic expression of C1r/s, MASPs, factor B/C2, C3, C4, C5, C6, C7, C8, C9, and factor I. A remarkable difference in the expression time course was noted between the isotypes in C3, C4, and C5. Especially, teleost-specific isotypes of C3 and C4 (non-histidine-type) started around hatching, in contrast to evolutionarily common isotypes (histidine-type), which showed much earlier expression. Whole-mount in situ hybridization of carp embryos revealed some difference in embryonic expression sites of two major C3 isotypes (C3-H1 and C3-S) in addition to common expression sites such as the yolk syncytial layer. The temporal and spatial differences in expression among the isotypes suggest that the isotypes are functionally differentiated in teleost early development.. |
| 88. | 辻倉正和, 中尾 実樹, 杣本 智軌, ゲノムデータからみた魚類補体制御因子の多様性, アクアゲノム研究会東京大会, 2012.03. |
| 89. | 多治見誠亮・杣本智軌・中西照幸・中尾実樹, 不活化ウイルス経腸管感作後のギンブナT細胞の免疫応答, 日本水産学会九州支部大会, 2012.01. |
| 90. | 徳永弓枝, 白水正道, 安住 薫, 吉浦康寿, 乙竹 充, 田代 康介, 杣本 智軌, 中尾 実樹, rag-1遺伝子欠損によるゼブラフィッシュの獲得免疫不全の検証と遺伝子の発現変動, 日本水産学会秋季大会, 2011.10. |
| 91. | 中尾 実樹, 補体の内外に見るホメオスタシス：活性化制御と視細胞クリアランス, 日本動物学会第82回大会シンポジウム「第１回ホメオスタシスバイオロジーシンポジウム」, 2011.09. |
| 92. | 辻倉正和, 杣本 智軌, 鵜木陽子, 中尾 実樹, 魚類における補体活性化制御因子の多様性, 第23回日本比較免疫学会, 2011.08. |
| 93. | 長沢貴宏, 中易千早, 松山知正, 杣本 智軌, 中尾 実樹, 魚類栓球の貪食作用とその活性化に伴う形態変化, 第23回日本比較免疫学会, 2011.08. |
| 94. | 白水 正道・徳永 弓枝・吉浦 康寿、乙竹 充・杣本 智軌・中尾 実樹, Rag1欠損ゼブラフィッシュにおける獲得免疫不全の検証, 第23回日本比較免疫学会, 2011.08. |
| 95. | Miki Nakao, Masakazu Tsujikura, Satoko Ichiki, Vo Kha Tam, Tomonori Somamoto, Structural and functional diversity of the complement system, an innate immune factor, in fish, 8th International Congress of Comparative Physiology and Biochemistry, ICCPB-Nagoya, 2011.06. |
| 96. | Miki Nakao, Satoko Ichiki, Masakazu Tsujikura, Vo Kha Tam, Tomonori Somamoto, Complement system in teleost fish: Isotypic diversity in pathogen-recognition, activation cascade, and ontogeny, Comparative Immunology and Pathology Workshop Edmonton 2011, 2011.05. |
| 97. | Takahiro Nagasawa, Chihara Nakayasu, Tomomasa Matsuyama, Tomonori Somamoto, Miki Nakao, Phagocytic activities of carp (Cyprinus carpio ) thrombocytes, Comparative Immunology and Pathology Workshop Edmonton 2011, 2011.05. |
| 98. | 長澤貴宏・松山知正・中易千早・杣本智軌・中尾実樹, 魚類栓球の異物貪食作用, 日本水産学会春季大会, 2011.03. |
| 99. | 一木昭土、塚本春香、杣本智軌、中尾実樹, コイ補体レクチン経路に関連するコレクチンの異物認識多様性, 第47回補体シンポジウム, 2010.09. |
| 100. | Nakao, M., Ichiki, S., Mutsuro, J., Tsujikura, M., and Somamoto, T., Functional diversity of the complement component isotypes in bony fish innate immunity: a model study using the common carp., 9th International Congress on the Biology of Fish, 2010.07. |
| 101. | 赤星佐和・辻倉正和・一木智子・杣本智軌・中尾実樹, コイ補体成分 C7 アイソタイプの機能分化, 日本水産学会春季大会, 2010.03. |
| 102. | 一木昭土, 塚本春香, 杣本 智軌, 中尾 実樹, コイ補体関連血清レクチンアイソタイプの異物認識多様性, 日本水産学会春季大会, 2010.03. |
| 103. | 中尾実樹, 硬骨魚類補体成分の高度な多様化と機能分化, 第46回補体シンポジウム, 2009.08. |
| 104. | 市居 敬、辻倉正和、杣本智軌、鵜木陽子、加藤愼一、吉国通庸、中尾実樹, コイC1複合体の亜成分組成, 第46回補体シンポジウム, 2009.08. |
| 105. | Vo Kha Tam、大蔵千恵、杣本智軌、中尾実樹, コイ初期発生におけるC3アイソタイプの発現パターン, 第46回補体シンポジウム, 2009.08. |
| 106. | 一木智子、鵜木 (加藤) 陽子、杣本智軌、中尾実樹, 魚類C3アイソタイプの生体防御における機能分化, 第46回補体シンポジウム, 2009.08. |
| 107. | 中村一規・杣本智軌・中西照幸・中尾実樹, ギンブナFas ligandのcDNAクローニングと発現解析, 日本比較免疫学会, 2009.08. |
| 108. | 長澤貴宏・中易千早・杣本智軌・中尾実樹, コイ栓球は異物を貪食する, 日本比較免疫学会, 2009.08. |
| 109. | 市居 敬・辻倉正和・杣本智軌・鵜木陽子・加藤愼一・吉国通庸・中尾実樹, コイ補体C1複合体を構築するサブユニットの同定, 日本比較免疫学会, 2009.08. |
| 110. | Soha Gomaa, Tomonori Somamoto and Miki Nakao, Cloning, purification and characterization of zymosan-binding proteins in Nile Tilapia (Oreochromis niloticus) , 日本比較免疫学会, 2009.08. |
| 111. | Miki Nakao, STRUCTURAL AND FUNCTIONAL DIVERSITY OF COMPLEMENT COMPONENTS IN BONY FISH: IMMUNOLOGICAL AND OTHER BIOLOGICAL IMPLICATIONS, The 2009 International Symposium on Fish Defense Mechanisms against Invading Pathogens, 2009.07. |
| 112. | Shinichi Urabe, Tomonori Somamoto, Shiro Sameshima, Teruyuki Nakanishi, Mitsuru Ototake, and Miki Nakao, Characterization of MHC class I genes in clonal ginbuna crucian carp, Carassius auratus langsdorfii, 11th Congress of International Society of Developmental and Comparative Immunology, 2009.06. |
| 113. | Satoko Ichiki, Yoko Kato-Unoki, Tomonori Somamoto, and Miki Nakao, FUNCTIONAL ANALYSIS OF CARP COMPLEMENT C3 ISOTYPES USING MONOCLONAL ANTIBODIES, 11th Congress of International Society of Developmental and Comparative Immunology, 2009.06. |
| 114. | Takashi Ichii, Tomonori Somamoto, and Miki Nakao, MOLECULAR ARCHITECTURE OF C1 COMPLEX, THE FIRST COMPLEMENT COMPONENT, IDENTIFIED FROM THE COMMON CARP, 11th Congress of International Society of Developmental and Comparative Immunology, 2009.06. |
| 115. | 中尾実樹・杣本智軌・辻倉正和・一木智子, 魚類における抗体依存的な補体活性化経路の構成成分, 第12回日本マリンバイオテクノロジー学会, 2009.05. |
| 116. | Satoko ICHIKI, Yoko KATO-UNOKI, Tomonori SOMAMOTO, and Miki NAKAO, Functional Analysis of Carp Complement C3 Isotypes Using Monoclonal Antibodies, World Fisheries Congress 2008, 2008.10. |
| 117. | Sawa AKAHOSHI, Tomonori SOMAMOTO, and Miki NAKAO, Identification and Expression Analysis of Two Isotypes of Carp Complement Component C7, World Fisheries Congress 2008, 2008.10. |
| 118. | Masakazu TSUJIKURA, Tomonori SOMAMOTO, Yoko KATO-UNOKI, and Miki NAKAO, Molecular Identification of a Novel Regulator of Complement Activation in Teleost Immune System, World Fisheries Congress 2008, 2008.10. |
| 119. | Miki NAKAO, Tomonori SOMAMOTO, Yoko KATO-UNOKI, and Junichi MUTSURO, The Complement System of Teleost: Isotypic Diversity of Its Components in the Structure, Expression, and Functions., World Fisheries Congress 2008, 2008.10. |
| 120. | Miki Nakao, Functional divergence and its biological significance of complement component isotypes in bony fish., Swedish-Japan STINT-meeting on Innate Immunity, 2008.10. |
| 121. | 占部慎二、鮫島史朗、杣本智軌、中西照幸、中尾実樹, ギンブナ由来細胞株における4種のMHCクラスI遺伝子のクローニングおよび、CHNV感染後の発現動態, 日本比較免疫学会, 2008.08. |
| 122. | 辻倉正和・杣本智軌・鵜木陽子・中尾実樹, コイおよびゼブラフィッシュの膜型補体制御因子, 日本比較免疫学会, 2008.08. |
| 123. | 一木昭土、畑中大作、杣本智軌、中尾実樹, 補体レクチン経路に関与するコイ血清レクチンの多様性, 日本比較免疫学会, 2008.08. |
| 124. | Miki Nakao, Junichi Mutsuro, Yoko Kato-Unoki, Tomoki Yano, and Alister W. Dodds., C3 and C4 isotypes with a non-catalyzed thioester in carp fish: phylogenetic implications, Immunochemistry in Oxford Symposium, 2008.07. |
| 125. | 赤星佐和、辻倉正和、杣本智軌、中尾実樹, コイ補体成分C7アイソタイプのクローニングと発現解析, 補体シンポジウム, 2008.07. |
| 126. | Miki Nakao, Junichi Mutsuro, Yoko Kato-Unoki, and Tomonori Somamoto, Structural and functional diversity of the complement components in bony fish: implication for innate immune defense., International Conference of Advanced Research on Marine Bioresources, 2008.05. |
| 127. | Miki Nakao, Junichi Mutsuro, Yoko Kato-Unoki, and Tomonori Somamoto, Structural and functional diversity of the complement components in bony fish: implication for innate immune defense, International Conference of Advanced Research on Marine Bioresources, 2008.05. |
| 128. | 杣本智軌・中西照幸・乙竹 充・中尾実樹, ウイルス特異的障害活性を有するギンブナ培養リンパ球のCD8αとTCRβのmRNAの発現解析, 日本水産学会, 2008.03. |
| 129. | 岩谷健太郎・杣本智軌・鵜木陽子・中尾実樹, コイ補体B因子アイソタイプ遺伝子の構造と発現制御, 日本水産学会, 2008.03. |
| 130. | 辻倉正和・杣本智軌・鵜木陽子・中尾実樹, 魚類における新規補体制御因子の同定, 日本水産学会, 2008.03. |
| 131. | 野中誠子･杣本智軌･中西照幸･乙竹充･中尾実樹, ２種類のギンブナCD4様分子, 日本水産学会九州支部会, 2008.01. |
| 132. | Miki Nakao, Antibacterial substances from bony fish: structure, function, and application, Philippines Society of Microbiology, 2007.10. |
| 133. | Satoko Ichiki, Yoko Kato-Unoki, Tomonori Somamoto, and Miki Nakao, Functional analysis of carp C3 isotypes using monoclonal antibodies, Japan-Germany International Cooperative Project on Education and Research, 2007.09. |
| 134. | Nevien K. Abdelkhalek, Asuka Komiya, Yoko Kato-Unoki, Tomonori Somamoto, Miki Nakao , Identification and expression analysis of a novel interleukine 8 (IL-8)-like CXC chemokine in carp (Cyprinus carpio), 日本魚病学会, 2007.09. |
| 135. | 鮫島史朗、鵜木陽子、杣本智軌、中尾実樹, ギンブナ頭腎、体腎、脾臓の Expressed Sequence Tag 解析, 日本比較免疫学会, 2007.08. |
| 136. | 杣本智軌、占部慎二、鮫島史朗、中西照幸、中尾実樹１, クローンギンブナ由来細胞株におけるMHCクラスＩによる抗原提示機構関連遺伝子のクローニング, 日本比較免疫学会, 2007.08. |
| 137. | 大蔵千恵、杣本智軌，近藤昌和，中尾実樹, コイの個体発生における補体成分アイソタイプの発現解析, 日本比較免疫学会, 2007.08. |
| 138. | 市居 敬、杣本 智軌、鵜木（加藤）陽子、中尾実樹, コイ補体成分C1タンパク質の同定, 補体シンポジウム, 2007.08. |
| 139. | 一木智子、鵜木（加藤）陽子、杣本智軌、中尾実樹, モノクローナル抗体を用いたコイC3アイソタイプの機能解析, 補体シンポジウム, 2007.08. |
| 140. | 辻倉正和、杣本智軌、鵜木（加藤）陽子、中尾実樹, 魚類における新規補体制御因子の同定, 補体シンポジウム, 2007.08. |
| 141. | 中尾実樹、加藤陽子、市居 敬、杣本智軌, コイ補体成分C4, C5の組換え体の発現と機能解析, 日本水産学会大会, 2007.03. |
| 142. | 新原美樹、杣本智軌、Vo Kha Tam、加藤陽子、中尾実樹, ゼブラフィッシュ補体成分の遺伝子の同定と発現解析, 日本水産学会大会, 2007.03. |
| 143. | 大蔵千恵、新原美樹、杣本智軌、中尾実樹、近藤昌和, 発現パターンから見た魚類補体の生体防御機能の推定, 南中九州・西四国水族防疫会議, 2007.02. |
| 144. | Yoko Kato-Unoki, Asuka Komiya, Shiro Sameshima, Miki Nakao, EST Analysis of mRNAs Expressed in Gill and Intestine of Carp (Cyprinus carpio. L), Asia-Pacific Aquatic Genomics Symposium 2006, 2006.11. |
| 145. | Miki Shinbara, Vo Kha Tam, Yoko Kato-Unoki, Tomonori Somamoto, and Miki Nakao, Identification of the genes encoding complement components from the zebrafish genome database, Asia-Pacific Aquatic Genomics Symposium 2006, 2006.11. |
| 146. | Miki Nakao, Taishi Yoshida, Jose P. Peralta and Aklani Rose D. Hidalgo, Molecular cloning of C3d, a fragment from complement component C3 with possible adjuvant activity, from marine cultured fish, International Forum on the Coastal Environment and Utitlization of Fisheries Resources, 2006.09. |
| 147. | 辻倉正和・杣本智軌・加藤陽子・中尾実樹 , ゼブラフィッシュ新奇補体制御因子のｃDNAクローニング, 日本魚病学会, 2006.09. |
| 148. | 吉田大志、杣本智軌，加藤陽子，中尾実樹, コイの新奇補体制御因子のcDNAクローニング, 日本比較免疫学会学術集会, 2006.08. |
| 149. | 杣本智軌、Vo Kha Tam、加藤陽子、中尾実樹, コイ補体C1q A,B,C鎖のcDNAのクローニング, 補体シンポジウム, 2006.08. |
| 150. | Chie Okura, Kentaro Iwatani, Daisuke Shibata, Yoko Kato-Unoki, Tomonori Somamoto and Miki Nakao, DIVESIFICATION OF COMPLEMENT COMPONENT ISOTYPES IN THE COMMON CARP: EXPRESSION PATTERN AND SOME FUNCTIONAL ASPECTS, 10th Congress of International Society for Developmental and Comparative Immunology, 2006.07. |
| 151. | Tomonori Somamoto, Seiko Nonaka, Yoko Kato, Mitsuru Ototake, Teruyuki Nakanishi and Miki Nakao, MOLECULAR CLONING AND CHARACTERIZATION OF CD4 IN GINBUNA CRUCIAN CARP, 10th Congress of International Society for Developmental and Comparative Immunology, 2006.07. |
| 152. | Tomonori Somamoto, Vo Kha Tam, Yoko Kato-Unoki and Miki Nakao, Molecular cloning and characterization of the complement C1q A, B, and C chains in common carp , 10th Congress of International Society for Developmental and Comparative Immunology, 2006.07. |
| 153. | Miki Nakao, THE COMPLEMENT SYSTEM IN INVERTEBRATES AND LOWER VERTEBRATES, 10th Congress of International Society for Developmental and Comparative Immunology, 2006.07. |
| 154. | Miki Nakao, JADCI: EVOLVING THROUGH ANNUAL MEETINGS, 10th Congress of International Society for Developmental and Comparative Immunology, 2006.07. |
| 155. | 新原美樹、Vo Kha Tam、杣本智軌、中尾実樹, ゼブラフィッシュゲノムデータベース中の補体成分遺伝子, 日本アクアゲノム研究会, 2006.04. |
| 156. | 柴田大輔、杣本智軌、中原マキ子、加藤陽子、中尾実樹, コイ補体制御因子の機能解析, 日本水産学会大会, 2006.04. |
| 157. | 中尾実樹, コイ科魚類を利用した魚類免疫研究の展開, 日本水産学会九州支部, 2005.10. |
| 158. | 小宮あすか、Christopher J.Bayne、加藤陽子、中尾実樹, ニジマスα2-マクログロブリンの多様性, 日本比較免疫学会学術集会, 2005.08. |
| 159. | 新原美樹、杣本智軌、Vo Kha Tam、加藤陽子、中尾実樹, ゼブラフィッシュ補体成分のin silico クローニング, 日本比較免疫学会学術集会, 2005.08. |
| 160. | 中尾実樹,吉田大志,小原浩平,藤本彩野,矢野友樹, コイ補体C3dフラグメントが抗体産生に及ぼす影響, 日本魚病学会, 2004.09. |
| 161. | 中尾実樹,原田あゆみ,矢野友樹, ソウギョ補体成分のcDNAクローニングとコイ補体遺伝子多重化機構の推定, 日本比較免疫学会, 2004.08. |
| 162. | 中尾実樹,加治屋貴之,畑中大作,中田宗宏,松下 操,藤田禎三,矢野友紀, コイ補体レクチン経路で機能する２種のMBL様レクチン, 補体シンポジウム, 2004.08. |
| 163. | Nakao M, Kajiya T, Hatanaka D, Matsushita M, Nakata M, Fujita T, Yano T, An MBL-MASP2 complex that functions in the lectin pathway of a bony fish, the common carp (Cyprinus carpio), International Complement Workshop, 2004.06. |
| 164. | 中尾実樹,加治屋貴之,前田 速,矢野友紀, コイ補体レクチン経路における Mannose-binding lectin familyの多様性, 日本水産学会, 2004.04. |
| 165. | 中尾実樹,前田 速,畑中大作,松下 操,中田宗宏,藤田禎三,矢野友紀, コイのマンノース結合レクチンの精製とクローニング, 日本比較免疫学会, 2003.08. |
| 166. | 片寄哲史,中尾実樹,松下 操,藤田禎三,矢野友紀, コイC1q様遺伝子のcDNAクローニング, 日本比較免疫学会, 2003.08. |
| 167. | Nakahara M,Harada A,Kondo K,Yano T,Nakao M, Diversity of the complement factor B isotypes in the common carp (Cyprinus carpio): Functional and genetic analysis, The 9th congress of the International Society of Developmental and Comparative Immunology, 2003.07. |
| 168. | Nakao M, Maeda H, Matsushita M, Nakata M, Fujita T, Yano T, Purification and cloning of the mannose-binding lectin (MBL) from the common carp (Cyprinus carpio), The 9th congress of the International Society of Developmental and Comparative Immunology, 2003.07. |
| 169. | Nakao M, Bayne CJ, Mutsuro J, Harada A, Bender RC, Yano T, Alpha-2-macroglobulin: Another diversified member of the thioester family of defence-related plasma proteins, The 9th congress of the International Society of Developmental and Comparative Immunology, 2003.07. |
| 170. | Hammond JA,Nakao M,Yano T,Smith VJ, Isolation of a novel GPI-anchored alpha-2-macroglobulin from Ciona intestinalis, The 9th congress of the International Society of Developmental and Comparative Immunology, 2003.07. |
| 171. | Nakao M,Nakahara M,Bayne CJ,Kondo M,Yano M, The complement factor B/C2-B lineage in teleost: A counterpart of mammalian C2?, The 9th congress of the International Society of Developmental and Comparative Immunology, 2003.07. |
| 172. | 中尾実樹,三浦知歌子,井藤俊亮,中原マキ子,奥村啓子,矢野友紀, コイ補体におけるC3dフラグメントの生成およびC3dレセプターの検出, 補体シンポジウム, 2003.07. |
| 173. | Nakao M, Hisamatsu S, Nakahara M, Kato Y, Smith SL, Yano T, Cloning of two factor I-isotypes with distinct domain organizations from the common carp, International Complement Workshop, 2002.10. |
| 174. | Nakao M, Nakahara M, Matsumoto M, Kato Y, Fujiki K, Yano T, Structural and functional diversity of factor B/C2-isotypes in the common carp, International Complement Workshop, 2002.10. |
| 175. | Nakao M, Diversity of innate immunity in bony fish: When, why and how carp have increased the genes coding for complement components ?, The 10th Internat. Joint Sem. "The future of agricultural science in Japan and Korea", 2002.10. |
| 176. | 中原マキ子,中尾実樹,無津呂淳一,矢野友紀, コイ補体B/C2アイソタイプの機能解析, 日本アクアゲノム研究会, 2002.09. |
| 177. | 中尾実樹,無津呂淳一,矢野友紀, 硬骨魚類における補体遺伝子の多重化とその意義, 日本アクアゲノム研究会, 2002.09. |
| 178. | 中尾実樹,久松里美,中原マキ子,無津呂淳一,加藤陽子,矢野友紀, コイ補体Ｉ因子アイソタイプの分子クローニング, 日本比較免疫学会学術集会, 2002.08. |
| 179. | 中尾実樹,中原マキ子,原田あゆみ,無津呂淳一,加藤陽子,矢野友紀, コイB因子/C2アイソタイプ遺伝子の連鎖解析および機能的多様性, 補体シンポジウム, 2002.07. |
| 180. | 中尾実樹,矢野友紀, サイトカインおよび補体の受容体, 日本水産学会, 2002.04. |
| 181. | 矢野友紀,中尾実樹, 魚類の補体系, 日本水産学会, 2002.04. |
| 182. | 中尾実樹, 魚類の補体系による防御戦略：補体成分の多様性, 日本比較３学会合同シンポジウム, 2001.12. |
| 183. | Nakao M, Mutsuro J, Yano T, The structural and functional diversity of complement components in carp, Cyprinus carpio, The 70th Anniversary of the Japanese Society of Fisheries Science International Commemorative Symposium, 2001.10. |
| 184. | 中原マキ子,中尾実樹,矢野友紀, コイ補体B因子/C2の組換えタンパク質の発現, 日本魚病学会, 2001.10. |
| 185. | 三浦知歌子,中尾実樹,無津呂淳一,矢野友紀, コイ補体C3からのC3d断片の生成, 日本魚病学会, 2001.10. |
| 186. | 加藤陽子,中尾実樹,無津呂淳一,矢野友紀, コイC5のcDNAクローニング, 補体シンポジウム, 2001.08. |
| 187. | 下薗真希,小玉 仁,森友忠明,中尾実樹,矢野友紀,苫名 充,中西照幸, コイIgMアイソフォームの好中球貪食に及ぼす効果, 日本生体防御学会, 2001.08. |
| 188. | 中尾実樹,無津呂淳一,田中則之,加藤陽子,矢野友紀, コイにおけるチオエステル含有タンパク質の多様性と進化, 日本比較免疫学会学術集会, 2001.07. |
| 189. | 中尾実樹,中原マキ子,Bayne CJ,藤木和浩,矢野友紀, 硬骨魚類の補体第２成分(C2)のcDNAクローニング, 日本水産学会, 2001.04. |
| 190. | Nakao M, Mutsuro J, Yano T, Diversity of complement components in bony fish., The 13th Annual Meeting of Japanese Association for Animal Cell Technology, 2000.11. |
| 191. | 中尾実樹,松元百恵,中沢美香,藤木和浩,矢野友紀, コイB因子/C2の多様性, 補体シンポジウム, 2000.08. |
| 192. | 三浦知歌子,中尾実樹,木村守孝,矢野友紀, 抗ペプチド抗体を用いたコイCR3βサブユニットの検出, 補体シンポジウム, 2000.08. |
| 193. | Nakao M, Matsumoto M, Nakazawa M, Fujiki K, Yano T, Diversity of the complement factor B/C2 in the common carp, 8th Congress of the International Society of Developmental and Comparative Immunology, 2000.07. |
| 194. | Nakao M, Osaka K, Kato Y, Yano T, Molecular cloning of C1r/C1s-like serine protease from the common carp, International Complement Workshop, 2000.07. |
| 195. | Nakao M, Osaka K, Kato Y, Yano T, Molecular cloning of C1r/C1s-like serine proteases of carp complement, 8th Congress of the International Society of Developmental and Comparative Immunology, 2000.07. |
| 196. | Kimura M, Nakao M, Miura C, Fujiki K, Yano T, Molecular cloning of a leukocyte integrin from the common carp, 8th Congress of the International Society of Developmental and Comparative Immunology, 2000.07. |
| 197. | Bayne CJ, Gerwick LG, Fujiki K, Nakao M, Yano T, Prospective acute phase (and other) proteins expressed in hepatocytes of rainbow trout, 8th Congress of the International Society of Developmental and Comparative Immunology, 2000.07. |
| 198. | Mutsuro J, Tanaka N, Totsuka S, Kato Y, Fujiki K, Nakao M, Yano T, Purification and cloning of two divergent C4 isotypes from the common carp, 8th Congress of the International Society of Developmental and Comparative Immunology, 2000.07. |
| 199. | Nakao M, Mutsuro J, Tanaka N, Totsuka S, Kato Y, Fujiki K, Tano T, Two divergent C4 isotypes from the common carp, International Complement Workshop, 2000.07. |
| 200. | 木村守孝,藤木和浩,中尾実樹,申 同浩,矢野友紀, コイ白血球インテグリンのcDNAクローニング, 日本比較免疫学会学術集会, 2000.07. |
| 201. | 中尾実樹,逢阪和則,加藤陽子,矢野友紀, コイ補体C1r/C1sのcDNAクローニング, 日本比較免疫学会学術集会, 2000.07. |
| 202. | 中尾実樹,下薗真希,小玉 仁,森友忠明,中西照幸,矢野友紀, コイIgMアイソフォームの精製と機能解析, 日本水産学会, 2000.04. |
| 203. | 中尾実樹,松元百恵,無津呂淳一,木村守孝,三浦知歌子,藤木和浩,矢野友紀, コイ補体B因子/C2の新規アイソタイプのcDNAクローニングおよび発現解析, 日本魚病学会, 2000.04. |
| 204. | 中尾実樹,逢阪和則,無津呂淳一,木村守孝,矢野友紀, コイ補体C1r/C1sのcDNAクローニング, 日本水産学会, 2000.04. |
| 205. | 田中則之,無津呂淳一,中尾実樹,矢野友紀, コイ補体C4アイソタイプ(C4A,C4B）のcDNAクローニング, 日本水産学会, 2000.04. |
| 206. | 加藤陽子,無津呂淳一,中尾実樹,矢野友紀, コイ補体第5成分(C5)に対するモノクローナル抗体の作成, 日本魚病学会, 2000.04. |
| 207. | 木村守孝,藤木和浩,中尾実樹,申 同浩,矢野友紀, コイC3レセプターβサブユニットのcDNAクローニングと遺伝子解析, 日本比較免疫学会学術集会, 1999.08. |
| 208. | 永井 毅,中尾実樹,加藤陽子,無津呂淳一,木村守孝,藤木和浩,矢野友紀, コイ補体MASPアイソフォームの分子クローニングおよび遺伝子解析, 日本比較免疫学会学術集会, 1999.08. |
| 209. | 無津呂淳一,中尾実樹,加藤陽子,藤木和浩,矢野友紀, コイ補体第３成分(C3)アイソフォームの単離・同定, 日本比較免疫学会学術集会, 1999.08. |
| 210. | Fujiki K, Shin DH, Nakao M, Yano T, Molecular cloning and expression analysis of carp (Cyprinus carpio) interleukin-1beta, high affinity immunoglobulin E Fc receptor gamma subunit and serum amyloid A., Ann. Internat. Symp. Fish. Soc. British Isles, 1999.07. |
| 211. | 中尾実樹,無津呂淳一,戸塚 悟,田中則之,加藤陽子,矢野友紀, コイC4の精製およびcDNAクローニング, 補体シンポジウム, 1999.06. |
| 212. | Kimura M, Fujiki K, Nakao M, Shin DH, Yano T, CR3 in bony fish: Molecular cloning of integrin beta 2-chain from carp, International Complement Workshop, 1998.10. |
| 213. | Nakao M, Mutsuro J,Obo J, Fujiki K, Yano T, Molecular cloning of multiple forms of carp C3, International Complement Workshop, 1998.10. |
| 214. | 藤木和浩,申 同浩,中尾実樹,矢野友紀, SSH法を用いたコイサイトカイン類のcDNAクローニング, 日本比較免疫学会学術集会, 1998.08. |
| 215. | 申 同浩,藤木和浩,中尾実樹,矢野友紀, コイnatural killer enhancing factorのcDNAクローニング, 日本比較免疫学会学術集会, 1998.08. |
| 216. | 中尾実樹,無津呂淳一,藤木和浩,矢野友紀, コイC3の多様性：複数のC3のcDNAクローニング, 補体シンポジウム, 1998.07. |
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