Development of system-wide zebrafish immune repertoire analysis at the individual level

• Project/Area Number: 15K14879
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Integrative animal science
• Research Institution: Hiroshima University
• Principal Investigator: TATSUO ICHINOHE 広島大学, 原爆放射線医科学研究所, 教授 (80314219)
• Co-Investigator(Kenkyū-buntansha): 本庶 仁子 広島大学, 原爆放射線医科学研究所, 講師 (80614106)
• Co-Investigator(Renkei-kenkyūsha): Suzuki Ryuji 国立病院機構, 相模原病院臨床研究センター・診断・治療室, 室長 (70373470)
• Research Collaborator: Kawase Takakazu ; Miyama Takahiko ; Sato Hiroyuki
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: ゼブラフィッシュ / 免疫系 / 主要組織適合性複合体 / 造血細胞移植 / 次世代シーケンサー / ゼブラフィッシュモデル / 主要組織適合性抗原 / 免疫学 / 高速シーケンシング / モデル動物

Outline of Final Research Achievements

In this study, we have developed methods enabling system-wide analysis of T cell receptor (TCR) repertoire at the individual level by taking advantage of a zebrafish model. We extended our analysis into human TCR repertoire and revealed that the immunodominant TCRs are shared by different individuals at least in the setting of anti-virus immune reaction and immune reconstitution after allogeneic hematopoietic cell transplantation.

Academic Significance and Societal Importance of the Research Achievements

従来、獲得免疫系を構成する細胞群を個別に同定することは可能であったが、それらの個体全体におけるレパトワを網羅的に取得する方法は存在しなかった。本研究では小型実験動物であるゼブラフィッシュの特性を活用し、システムワイドにゼブラフフィッシュの個体に存在するT細胞レパトワをα鎖、β鎖とも高頻度クローンから微小クローンまで決定する新規技術を確立した。この成果は、免疫系全体の動態を鳥瞰することを可能とし、今後、感染症や悪性腫瘍に対する免疫応答の解明や、免疫学的な機序により発生する病態の診断や治療法の開発に大きく貢献する。

Research Products

• [Journal Article] Next-Generation Immune Repertoire Sequencing as a Clue to Elucidate the Landscape of Immune Modulation by Host-Gut Microbiome Interactions (2018)
  • Author(s): Ichinohe Tatsuo、Miyama Takahiko、Kawase Takakazu、Honjo Yasuko、Kitaura Kazutaka、Sato Hiroyuki、Shin-I Tadasu、Suzuki Ryuji
  • Journal Title: Frontiers in Immunology Volume: 9 Pages: 668-668
  • DOI: 10.3389/fimmu.2018.00668
  • Peer Reviewed / Open Access
• [Journal Article] Highly functional T-cell receptor repertoires are abundant in stem memory T cells and highly shared among individuals (2017)
  • Author(s): Miyama Takahiko、Kawase Takakazu、Kitaura Kazutaka、Chishaki Ren、Shibata Masashi、Oshima Kumi、Hamana Hiroshi、Kishi Hiroyuki、Muraguchi Atsushi、Kuzushima Kiyotaka、Saji Hiroh、Shin-I Tadasu、Suzuki Ryuji、Ichinohe Tatsuo
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 668-668
  • DOI: 10.1038/s41598-017-03855-x
  • Peer Reviewed / Open Access
• [Journal Article] Tumor growth limited to subcutaneous site vs tumor growth in pulmonary site exhibit differential effects on systemic immunities. (2017)
  • Author(s): Masuda J, Takayama E, Strober W, Satoh A, Morimoto Y, Honjyo Y, Ichinose T, Tokuno SI, Ishizuka T, Nakata T, Mizutani A, Umemura N, Kitani A, Fuss IJ, Shigehiro T, Kawaki H, Mizuno-Kamiya M, Kondoh N, Seno M.
  • Journal Title: Oncol Rep Volume: 38 Issue: 1 Pages: 449-455
  • DOI: 10.3892/or.2017.5646
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 新しい養子免疫療法を目指したTALEN法によるT細胞受容体ゲノム編集技術の開発. (2017)
  • Author(s): 本庶仁子, 一戸辰夫.
  • Journal Title: 無菌生物. Volume: 47 Pages: 68-69
  • Peer Reviewed
• [Journal Article] Highly functional T-cell receptor repertoires are abundant in stem memory T cells and highly shared among individuals (2017)
  • Author(s): Takahiko Miyama, Takakazu Kawase, Kazutaka Kitaura, Ren Chishaki, Masashi Shibata, Kumi Oshima, Hiroshi Hamana, Hiroyuki Kishi, Atsushi Muraguchi, Kiyotaka Kuzushima, Hiroh Saji, Tadasu Shini, Ryuji Suzuki, Tatsuo Ichinohe
  • Journal Title: Scientific Reports Volume: in press
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] ゼブラフィッシュ初期胚における放射線応答の解析. (2016)
  • Author(s): 本庶仁子, 神谷研二, 一戸辰夫.
  • Journal Title: 広島医学 Volume: 69 Pages: 305-306
  • Peer Reviewed
• [Journal Article] Basic understanding of the HLA system in allogeneic hematopoietic cell transplantation (2015)
  • Author(s): 一戸辰夫.
  • Journal Title: Rinsho Ketsueki Volume: 56 Issue: 10 Pages: 2134-2143
  • DOI: 10.11406/rinketsu.56.2134
  • NAID: 130005103283
  • ISSN: 0485-1439, 1882-0824
  • Peer Reviewed
• [Presentation] 同種造血幹細胞移植後の末梢血免疫再構築におけるshared TCRの優位性. (2017)
  • Author(s): 美山貴彦, 川瀬孝和, 田中清人, 柴田真志, 樗木錬, 坂本葵, 土石川佳世, 森岡健彦, 大島久美, 本庶仁子, 田中秀則, 北浦一孝, 鈴木隆二, 一戸辰夫.
  • Organizer: 第39回日本造血細胞移植学会総会
  • Place of Presentation: 松江市
  • Year and Date: 2017-03-03
• [Presentation] 次世代シーケンサーとsingle cell sortingを用いた同種抗原反応性T細胞の網羅的解析と高頻度クローンの同定. (2017)
  • Author(s): 川瀬孝和, 坂本葵, 樗木錬, 美山貴彦, 柴田真志, 田中清人, 北浦一孝, 大島久美, 浜名洋, 岸裕幸, 葛島清隆, 田中秀則, 鈴木隆二, 一戸辰夫.
  • Organizer: 第39回日本造血細胞移植学会総会
  • Place of Presentation: 松江市
  • Year and Date: 2017-03-03
• [Presentation] T-cell receptor transgenic primary T cells using TALEN-mediated TCR gene editing as a novel tool to correct immunodeficiemcy caused by radiation damage. (2017)
  • Author(s): Takahiko Miyama, Yasuko Honjo, Takakazu Kawase, Tetsushi Sakuma, Takashi Yamamoto, Tatsuo Ichinohe.
  • Organizer: The 1st International Symposium of the network-type Joint Usage/Research Center for Radiation Disaster Medical Science-Scientific Underpinning for Restoration from a Radiation Diaster
  • Place of Presentation: 広島市
  • Year and Date: 2017-02-21
  • Int'l Joint Research
• [Presentation] T-cell receptor gene editing by transcription activator-like effector nuclease (TALEN) as a novel tool for adoptive T-cell immunotherapy. (2017)
  • Author(s): Yasuko Honjo, Takahiko Miyama, Takakazu Kawase, Tetsushi Sakuma, Takashi Yamamoto, Tatsuo Ichinohe.
  • Organizer: The 19th International Symposium on Gnotobiology
  • Int'l Joint Research
• [Presentation] Next-generation sequencing-based immunobioinformatics: toward a comprehensive understanding of the human immune system. (2017)
  • Author(s): Tatsuo Ichinohe.
  • Organizer: The 19th International Symposium on Gnotobiology
  • Int'l Joint Research / Invited
• [Presentation] Landscape of immune reconstitution after allogeneic hematopoietic cell transplantation is superdominated by distinct T cell populations bearing T-cell receptor clonotypes shared among different individuals. (2017)
  • Author(s): Takahiko Miyama, Takakazu Kawase, Yasuko Honjo, Kazutaka Kitaura, Tadasu Shin-I, Ryuji Suzuki, Tatsuo Ichinohe.
  • Organizer: 59th Annual Meeting of the American Society of Hematology
  • Int'l Joint Research
• [Presentation] ゼブラフィッシュ初期胚の放射線に対する影響の解析. (2016)
  • Author(s): 本庶仁子, 一戸辰夫.
  • Organizer: 日本放射線影響学会第59回大会
  • Place of Presentation: 広島市
  • Year and Date: 2016-10-26
• [Presentation] 次世代シーケンサーと single cell sorting を用いた T 細胞受容体 (TCR) の 網羅的解析. (2016)
  • Author(s): 川瀬孝和, 美山貴彦, 一戸辰夫.
  • Organizer: 第25回日本組織適合性学会大会
  • Place of Presentation: 札幌市
  • Year and Date: 2016-10-23
  • Invited
• [Presentation] サイトメガロウイルス反応性T細胞レパトワ形成に与えるHLA-A*02の影響の次世代シーケンサーを用いた網羅的解析. (2016)
  • Author(s): 美山貴彦, 田中清人, 柴田真志, 川瀬孝和, 樗木錬, 坂本葵, 北浦一孝, 大島久美, 浜名洋, 岸裕幸, 葛島清隆, 田中秀則, 鈴木隆二, 一戸辰夫.
  • Organizer: 第25回日本組織適合性学会大会
  • Place of Presentation: 札幌市
  • Year and Date: 2016-10-23
• [Presentation] Stem cell memory T-cells are a reservoir of functional T-cells highly shared among individuals. (2016)
  • Author(s): Takahiko Miyama, Takakaza Kawase, Kazutaka Kitaura, Ren Chishaki, Masashi Shibata, Kumi Oshima, Hiroshi Hamana, Hiroyuki Kishi, Kiyotaka Kuzushima, Hiroh Saji, Ryuji Suzuki, Tatsuo Ichinohe.
  • Organizer: 第78回日本血液学会学術集会
  • Place of Presentation: 横浜市
  • Year and Date: 2016-10-14
• [Presentation] 次世代シーケンサーとsingle cell sortingを用いた同種抗原反応性T細胞の網羅的解析. (2016)
  • Author(s): 川瀬孝和, 坂本葵, 樗木錬, 美山貴彦, 柴田真志, 田中清人, 北浦一孝, 大島久美, 浜名洋, 岸裕幸, 葛島清隆, 佐治博夫, 鈴木隆二, 一戸辰夫.
  • Organizer: 第20回日本がん免疫学会総会
  • Place of Presentation: 大阪市
  • Year and Date: 2016-07-28
• [Presentation] 同種造血幹細胞移植後のCD4+およびCD8+T細胞再構築過程の次世代シーケンサーを用いた網羅的解析. (2016)
  • Author(s): 大島久美, 北浦一孝, 美山貴彦, 川瀬孝和, 井上暢子, 山田恭平, 粟屋忠佑, 石田誠子, 森岡健彦, 今川潤, 三原圭一朗, 中十奈苗, 鈴木隆二, 一戸辰夫.
  • Organizer: 第38回日本造血細胞移植学会総会.
  • Place of Presentation: 名古屋市
  • Year and Date: 2016-03-04
• [Presentation] Comprehensive T-cell receptor repertoire analysis using deep sequencing and single cell cloning reveals extreme oligoclonality of ex vivo expanded cytomegalovirus-reactive cytotoxic T-cells. (2015)
  • Author(s): Takahiko Miyama, Takakazu Kawase, Kazutaka Kitaura, Ren Chishaki, Masashi Shibata, Kumi Oshima, Hiroshi Hamana, Hiroyuki Kishi, Kiyotaka Kuzushima, Hiroh Saji, Ryuji Suzuki, Tatsuo Ichinohe.
  • Organizer: 57th Annual Meeting of the American Society of Hematology
  • Place of Presentation: Orlando, FL, U.S.A.
  • Year and Date: 2015-12-05
  • Int'l Joint Research
• [Presentation] Comprehensive analysis of CMV-specific T-cell repertoire by deep sequencing and single cell cloning. (2015)
  • Author(s): Takahiko Miyama, Takakazu Kawase, Kazutaka Kitaura, Ren Chishaki, Masashi Shibata, Kumi Oshima, Hiroshi Hamana, Hiroyuki Kishi, Kiyotaka Kuzushima, Hiroh Saji, Ryuji Suzuki, Tatsuo Ichinohe.
  • Organizer: 第77回日本血液学会学術集会.
  • Place of Presentation: 金沢市
  • Year and Date: 2015-10-17
• [Presentation] Quantitative tracing of T-cell diversity using deep sequencing in a recipient of allogeneic HCT. (2015)
  • Author(s): Kumi Oshima, Kazutaka Kitaura, Takahiko Miyama, Takakazu Kawase, Tomoko Inoue, Seiko Ishida, Kyohei Yamada, Naoya Yamasaki, Tadasuke Awaya, Moe Okuno, Tetsumi Yoshida, Seiji Saito, Yoshiaki Kuroda, Sayaka Sugihara, Jun Imagawa, Shinya Katsutani, Keichiro Mihara, Teruhisa Fujii, Ryuji Suzuki, Tatsuo Ichinohe.
  • Organizer: 第77回日本血液学会学術集会.
  • Place of Presentation: 金沢市
  • Year and Date: 2015-10-16
• [Presentation] 次世代シーケンサーを用いた同種造血幹細胞移植後末梢血T細胞レパトワの網羅的解析. (2015)
  • Author(s): 大島久美, 北浦一孝, 美山貴彦, 川瀬孝和, 中十奈苗, 鈴木隆二, 一戸辰夫.
  • Organizer: 第24回日本組織適合性学会大会.
  • Place of Presentation: 水戸市
  • Year and Date: 2015-09-10
• [Presentation] Analysis of radiation-induced responses during early embryonic development of zebrafish. (2015)
  • Author(s): Yasuko Honjo, Kenji Kamiya, Tatsuo Ichinohe.
  • Organizer: 15th International Congress of Radiation Research
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2015-05-27
  • Int'l Joint Research
• [Remarks] ウイルスに対する反応性が高いＴ細胞は「共有Ｔ細胞レセプター」を利用していることを解明しました
  • URL: https://www.hiroshima-u.ac.jp/rbm/news/40259
• [Remarks] 原爆放射線医科学研究所 研究・教員紹介 血液・腫瘍内科
  • URL: https://www.hiroshima-u.ac.jp/rbm/research/lab/Hematology_and_Oncology
• [Remarks] 広島大学原爆放射線医科学研究所 血液・腫瘍内科研究分野 研究紹介
  • URL: http://home.hiroshima-u.ac.jp/hematol/research-activities.html
• [Patent(Industrial Property Rights)] Platinum TALENを用いたT細胞受容体の完全置換技術 (2017)
  • Inventor(s): 一戸辰夫、山本 卓、佐久間哲史、本庶仁子、他３名
  • Industrial Property Rights Holder: 一戸辰夫、山本 卓、佐久間哲史、本庶仁子、他３名
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2017-197010
  • Filing Date: 2017
• [Patent(Industrial Property Rights)] エフェクターT細胞(Teff)抗原受容体を用いた抗原特異的制御性T 細胞(Treg)の作出技術 (2017)
  • Inventor(s): 一戸辰夫、山本 卓、佐久間哲史、本庶仁子、他３名
  • Industrial Property Rights Holder: 一戸辰夫、山本 卓、佐久間哲史、本庶仁子、他３名
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2017-197010
  • Filing Date: 2017