Development of in vivo cytotoxicity assay system using humanized mice increasing human natural killer cells

• Project/Area Number: 25871075
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Laboratory animal science; Tumor therapeutics
• Research Institution: Central Institute for Experimental Animals
• Principal Investigator: Katano Ikumi 公益財団法人実験動物中央研究所, その他部局等, 研究員 (90442558)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: ヒト化マウス / NK細胞 / ヒト免疫 / ｈIL-2 / hIL-15 / hIL-2

Outline of Final Research Achievements

We previously reported that human NK cells are predominately developed in NOG-hIL-2 Tg mice after human hematopoietic stem cell (HSC) transplantation. In this study, we assessed in vivo antibody-dependent-cellular-cytotoxicity (ADCC) activity using human HSC-transferred NOG-hIL-2 Tg mice engrafted with CCR4 expressing Hodgkin’s lymphoma cell line L428. After treatment with an anti-CCR4 antibody, tumor growth was slightly suppressed in NOG-hIL-2 Tg mice. To improve the function of NK cells, we generated a NOG-hIL-15 Tg mouse. Although human mature NK cells were dominantly differentiated in NOG-hIL-15 Tg mice, they immediately died within 8 weeks after HSC transplantation. In contrast, human peripheral blood-derived mature NK cells succeeded to be maintained over 3 months after transplantation without death. Moreover, these NK cells showed in vivo cytotoxic activity against engrafted K562 cells. Therefore, NOG-hIL-15 Tg mice are useful tools for investigation of in vivo NK cytotoxicity.

Research Products

• [Journal Article] Predominant Development of Mature and Functional Human NK Cells in a Novel Human IL-2-Producing Transgenic NOG Mouse (2015)
  • Author(s): Katano, I. Takahashi, T. Ito, R. Kamisako, T. Mizusawa, T. Ka, Y. Ogura, T. Suemizu, H. Kawakami, Y. Ito, M.
  • Journal Title: J Immunol Volume: 194 Issue: 7 Pages: 3513-3525
  • DOI: 10.4049/jimmunol.1401323
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Visualization of the human CD4(+) T-cell response in humanized HLA-DR4-expressing NOD/Shi-scid/gammac(null) (NOG) mice by retrogenic expression of the human TCR gene (2015)
  • Author(s): Takahashi, T. Katano, I. Ito, R. Ito, M.
  • Journal Title: Biochem Biophys Res Commun Volume: 456 Issue: 1 Pages: 219-224
  • DOI: 10.1016/j.bbrc.2014.11.062
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] NOD-Rag2null IL-2Rγnull mice: an alternative to NOG mice for generation of humanized mice. (2014)
  • Author(s): Katano I, Ito R, Kamisako T, Eto T, Ogura T, Kawai K, Suemizu H, Takahashi T, Kawakami Y, Ito M.
  • Journal Title: Exp Anim Volume: 63 Pages: 321-330
  • NAID: 130004677826
  • Peer Reviewed / Open Access
• [Journal Article] A novel enhanced green fluorescent protein-expressing NOG mouse for analyzing the microenvironment of xenograft tissues. (2014)
  • Author(s): Higuchi Y, Kawai K, Yamamoto M, Kuronuma M, Ando Y, Katano I, Nakamura M, Suemizu H.
  • Journal Title: Exp Anim Volume: 63 Pages: 55-62
  • Peer Reviewed
• [Journal Article] Establishment of a Human Allergy Model Using Human IL-3/GM-CSF-Transgenic NOG Mice. (2013)
  • Author(s): Ito, R., T. Takahashi, I. Katano, K. Kawai, T. Kamisako, T. Ogura, M. Ida-Tanaka, H. Suemizu, S. Nunomura, C. Ra, A. Mori, S. Aiso, and M. Ito.
  • Journal Title: J Immunol Volume: 191 Issue: 6 Pages: 2890-2899
  • DOI: 10.4049/jimmunol.1203543
  • Peer Reviewed
• [Presentation] Human interleukin-15 transgenic NOG mice support the long-term maintenance of human mature NK cells from peripheral blood (2016)
  • Author(s): Ikumi Katano, Takeshi Takahashi, Ryoji Ito, Asami Hanazawa, Mamoru Ito
  • Organizer: 5th International Workshop on Humanized Mice (IWHM5)
  • Place of Presentation: Switzerland, University of Zurich
  • Year and Date: 2016-01-28
  • Int'l Joint Research
• [Presentation] Human interleukin-15 transgenic NOG mice support the long-term maintenance of human mature NK cells from peripheral blood (2015)
  • Author(s): Ikumi Katano, Takeshi Takahashi, Ryoji Ito, Asami Hanazawa, Yutaka Kawakami, Mamoru Ito
  • Organizer: 第４４回日本免疫学会学術集会
  • Place of Presentation: 札幌、札幌コンベンションセンター
  • Year and Date: 2015-11-18
• [Presentation] ヒトIL-15遺伝子導入NOGマウスはヒト末梢血由来成熟NK細胞を長期間維持できる (2015)
  • Author(s): 片野いくみ、高橋武司、伊藤亮治、花澤麻美、高橋利一、末水洋志、河上裕、伊藤守
  • Organizer: 第62回日本実験動物学会総会
  • Place of Presentation: 京都テルサ
  • Year and Date: 2015-05-28 – 2015-05-30
• [Presentation] ヒトIL-15遺伝子導入NOGマウスはヒト末梢血由来成熟NK細胞を長期間維持できる (2015)
  • Author(s): 片野いくみ、高橋武司、伊藤亮治、花澤麻美、高橋利一、末水洋志 、河上裕、伊藤守
  • Organizer: 第６２回日本実験動物学会総会
  • Place of Presentation: 京都、京都テルサ
  • Year and Date: 2015-05-27
• [Presentation] Characterization of human Eosinophil in HSC-transferred NOG-human IL-5 transgenic mice (2013)
  • Author(s): Ikumi Katano, Takeshi Takahashi, Ryoji Ito, Mamoru Ito
  • Organizer: 第42回日本免疫学会学術集会
  • Place of Presentation: 千葉・幕張メッセ
• [Presentation] Predominant Development of Mature and Functional Human NK Cells in a Novel Human IL-2 Producing Transgenic NOD-scid,IL-2Rg KO (NOG) Mouse (2013)
  • Author(s): Ikumi Katano, Takeshi Takahashi, Ryoji Ito, Mamoru Ito
  • Organizer: The 4th International Workshop on Humanized Mice (IWHM4)
  • Place of Presentation: 韓国・ソウル
• [Presentation] Functional NK cells developed from human hematopoietic stem cell in human interleukin-2 transgenic NOG mice (2013)
  • Author(s): Ikumi Katano, Ryoji Ito, Takeshi Takahashi, Mamoru Ito
  • Organizer: 15th International Congress of Immunology
  • Place of Presentation: イタリア・ミラノ(Mico)
• [Patent(Industrial Property Rights)] ヒトIL-15分泌免疫不全マウス (2015)
  • Inventor(s): 伊藤守、片野いくみ
  • Industrial Property Rights Holder: 伊藤守、片野いくみ
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2015-107932
  • Filing Date: 2015-05-27