The determination of innate immune responses using cytokine genes in fish

• Project/Area Number: 23580257
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General fisheries
• Research Institution: University of Miyazaki
• Principal Investigator: SAKAI masahiro 宮崎大学, 農学部, 教授 (20178536)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2013: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2011: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
• Keywords: サイトカイン / フグ / 自然免疫 / プロバイオティクス / Multiplex RT-PCR / インターフェロン / マルチプレックスPCR / 免疫賦活剤

Research Abstract

To monitor the expression of cytokine genes in Japanese pufferfish, a novel platform for quantitative multiplexed analysis was developed. This custom-designed multiplex RT-PCR assay was used to analyze the expression profiles of 19 cytokine genes, including pro-inflammatory, anti-inflammatory, T-cell proliferation/differentiation, B-cell activation/differentiation, NK cell stimulation, induction of anti-viral activity, and monocyte/macrophage progenitor cell proliferation cytokines in head kidney cells under immune stimulatory conditions. The expression profiles were dissimilar in the unstimulated control and immune-stimulated cells. Moreover, increased expression profile was observed due to different stimulations for IL-1b, IL-6, IL-10, IL-12p35, IL-12p40, IL-21, TNF-a, TNF-N, I-IFN-1 and IFN-g genes. These results suggest that cytokine genes could be used as biomarkers to know the innate immune status of fish.

Research Products

• [Journal Article] Establishment of a multiplex RT-PCR assay for the rapid detection of fish cytokines (2013)
  • Author(s): Kono, T., Takayama, H., Nagamine, R., Korenaga, H., Sakai, M.
  • Journal Title: Veterinary Immunology and Immunopathology Volume: 151 Pages: 90-101
  • Peer Reviewed
• [Journal Article] Cytokine responses in the Japanese pufferfish (Takifugu rubripes) head kidney cells induced with heat-killed probiotics isolated from the Mongolian dairy products (2013)
  • Author(s): Biswas, G., Korenaga, H., Nagamine, R., Takayama, H., Kawahara, S., Takeda, S., Kikuchi, Y., Dashnyam, B., Kono, T., Sakai, M.
  • Journal Title: Fish and Shellfish Immunology Volume: 34 Pages: 1170-1177
  • Peer Reviewed
• [Journal Article] Cytokine mediated immune responses in the Japanese pufferfish (Takifugu rubripes) administered with heat-killed Lactobacillus paracasei spp. paracasei (06TCa22) isolated from the Mongolian dairy product (2013)
  • Author(s): Biswas, G., Korenaga, H., Nagamine, R., Kawahara, S., Takeda, S., Kikuchi, Y., Dashnyam, B., Yoshida, T., Kono, T., Sakai, M.
  • Journal Title: International Immunopharmacology Volume: 17 Pages: 358-365
  • Peer Reviewed
• [Journal Article] Elevated cytokine responses to Vibrio harveyi infection in the japanese pufferfish (Takifugu rubripes) treated with lactobacillus paracasei spp. paracasei (06TCa22) isolated from the mongolian dairy product (2013)
  • Author(s): Biswas, G., Korenaga, H., Nagamine, R., Kawahara, S., Takeda, S., Kikuchi, Y., Dashnyam, B., Yoshida, T., Kono, T., Sakai, M.
  • Journal Title: Fish and Shellfish Immunology Volume: 35 Pages: 756-765
  • Peer Reviewed
• [Journal Article] Cytokine mediated immune responses in the Japanese pufferfish (Takifugu rubripes) administered with heat-killed Lactobacillus paracasei spp. paracasei (06TCa22) isolated from the Mongolian dairy product. (2013)
  • Author(s): Biswas G., Korenaga H., Nagamine R., Kawahara S., Takeda S., Kikuchi Y., Dashnyam B., Yoshida T., Kono T. and Sakai M.
  • Journal Title: International Immunopharmacology Volume: 17 Issue: 2 Pages: 358-365
  • DOI: 10.1016/j.intimp.2013.06.030
  • Peer Reviewed
• [Journal Article] Elevated cytokine responses to vibrio harveyi infection in the japanese pufferfish (Takifugu rubripes) treated with Lactobacillus paracasei spp. paracasei (06TCa22) isolated from the mongolian dairy product. (2013)
  • Author(s): Biswas G., Korenaga H., Nagamine R., Kawahara S., Takeda S., Kikuchi Y., Dashnyam B., Yoshida T., Kono T. and Sakai M.
  • Journal Title: Fish and Shellfish Immunology Volume: 35 Issue: 3 Pages: 756-765
  • DOI: 10.1016/j.fsi.2013.06.004
  • Peer Reviewed
• [Journal Article] Cytokine responses in the Japanese pufferfish (Takifugu rubripes) head kidney cells induced with heat-killed probiotics isolated the Mongolian dairy products (2013)
  • Author(s): Biswas, G., Korenaga, H., Nagamine, R., Takayama, H., Kawahara, S., Takeda, S., Kikuchi, Y., Dashnyam, B., Kono, T., Sakai, M.
  • Journal Title: Fish and shellfish Immunology Volume: 34 Pages: 1170-1177
  • Peer Reviewed
• [Journal Article] サイトカイン遺伝子を用いた魚類免疫賦活剤の効果判定 (2012)
  • Author(s): 酒井正博・高山博章・河野智哉
  • Journal Title: 養殖 Volume: 613 Pages: 52-57
• [Presentation] ホタテ貝およびアワビの煮術で処理したトラフグ頭腎細胞におけるサイトカイン遺伝子の発現 (2013)
  • Author(s): 加藤翔太、是永大樹、引間順一、河野智哉、酒井正博
  • Organizer: 平成25年度日本水産学会秋季大会
  • Place of Presentation: 津
  • Year and Date: 2013-09-19
• [Presentation] A breakthrough in identifying neuropeptide using"reverse pharmacology"from fish (2013)
  • Author(s): Korenaga, H., Hamasuna, S., Ida, T., Sakai, M., Nagamine, R., Kono, T.
  • Organizer: 1^<st> International Conference of Fish and Shellfish Immunology
  • Place of Presentation: ビーゴ、スペイン
  • Year and Date: 2013-06-25
• [Presentation] A brekthrough in identifying neuropeptide using reverse pharmacology from fish (2013)
  • Author(s): korenaga, H., Hamasuna, S., Ida, T., Sakai, M., Nagamine, R., Kono, T.
  • Organizer: Fish and Shellfish Immunology
  • Place of Presentation: Vigo, Spain
• [Presentation] トラフグ単球におけるサイトカイン遺伝子の発現プロファイル (2013)
  • Author(s): 河野智哉、長峰隆介、是永大樹、酒井正博
  • Organizer: 平成２５年度日本水産学会秋季大会
  • Place of Presentation: 津
• [Presentation] A multiplex RT-PCR assay for rapid and sensitive detection of cytokines in fish (2012)
  • Author(s): Kono, T., Takayama, H., Nagamine, R., Korenaga, H., Sakai, M.
  • Organizer: 10^<th> Joint Annual Meeting of International Cytokine Society and International Society for Interferon and Cytokine Research
  • Place of Presentation: ジュネーブ、スイス
  • Year and Date: 2012-09-11
• [Presentation] A multiplex RT-PCR assay for rapid and sensitive detection of cytokines in fish (2012)
  • Author(s): Kono, T., Takayama, H., Nagamine, R., Korenaga, H., Sakai, M.
  • Organizer: 10th Joint Meeting of international Cytokine Society and International Society for Interferon and Cytokine Research
  • Place of Presentation: Geneva, Switzerland
• [Presentation] LPS等で処理したフグ頭腎細胞の サイトカイン遺伝子の発現定量解析 (2012)
  • Author(s): 高山博章・是永大樹・酒井正博・長峰隆介・河野智哉
  • Organizer: 平成２４年度日本魚病学会春季大会
  • Place of Presentation: 東京海洋大学
• [Presentation] The expression analysis of fugu cytokine genes by mulyiplex RT-PCR assay (2011)
  • Author(s): Takayama, H., Kuze, H., Korenaga, H., Sakai, M., Kono, T.
  • Organizer: The 15^<th> EAFP
  • Place of Presentation: スプリット、クロアチア
  • Year and Date: 2011-09-12
• [Remarks]
  • URL: http://www.agr.miyazaki-u.ac.jp/~abs/index.html