腫瘍細胞死誘導時における腫瘍貪食樹状細胞のＣＤ８+Ｔ細胞活性化機構の可視化解明

• Project Area: Mutli-dimensional fluorescence live imaging of cellular function and molecular activity
• Project/Area Number: 25113711
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Kyoto University
• Principal Investigator: 戸村 道夫 京都大学, 医学(系)研究科(研究院), 准教授 (30314321)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥10,660,000 (Direct Cost: ¥8,200,000、Indirect Cost: ¥2,460,000); Fiscal Year 2014: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2013: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
• Keywords: イメージング / ガン / 樹状細胞 / T細胞 / 生体内イメージング / 免疫系 / 化学療法 / 抗原提示細胞

Outline of Annual Research Achievements

強力な抗腫瘍免疫応答の誘導は腫瘍克服に繋がる重要なテーマである。抗がん剤治療時には大量の細胞死した腫瘍細胞が供給されるため、腫瘍内浸潤樹状細胞(DC)の細胞死した腫瘍の貪食をはじめ、所属リンパ節における抗原特異的CD8+T細胞との相互作用は大きく変化すると考えられる。従って、その解明は抗がん剤治療時における抗腫瘍免疫誘導を考える上で重要である。そこで本研究は、腫瘍細胞死誘導時における、腫瘍内浸潤DCの腫瘍細胞貪食、所属リンパ節への移行、そして、DCと抗原特異的CD8+T細胞の所属リンパ節における相互作用をreal time可視化により明らかにすることを目的とした。
腫瘍内に浸潤したDCを赤色にマーキングすることで、腫瘍から所属リンパ節に移行した腫瘍浸潤DCを追跡･可視化出来る、紫光により緑から赤色に変色する光変換蛍光タンパク質KikGRを樹状細胞系列特異的に発現するマウスに、蛍光タンパク質tdKeima、モデル抗原、および細胞死誘導のためにDiphtheria toxin-receptorを発現させた腫瘍細胞株を移植して用いた。
腫瘍細胞内には沢山のKikGR陽性DCが観察されたが、腫瘍の貪食は殆ど認められなかった。しかし、腫瘍細胞死を誘導すると、殆どの腫瘍内浸潤DCが腫瘍細胞を貪食し、腫瘍浸潤DCの所属リンパ節への移行も増加した。そして、腫瘍から所属リンパ節に移行した赤色にマーキングしたDCと抗原特異的CD8+T細胞の相互作用も観察された。そこでさらに、腫瘍細胞死の誘導とともにDCの成熟を促進するLPSを腫瘍内に投与すると抗原特異的CD8+T細胞の増殖促進が認められた。以上の結果から、抗がん剤治療時に腫瘍内浸潤DCの成熟促進を組み合わせることにより、より強い抗腫瘍免疫応答を誘導できる可能性が示された。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Real-time tracking of cell cycle progression during CD8(+) effector and memory T-cell differentiation. (2015)
  • Author(s): Kinjyo I, Qin J, Tan SY, Wellard CJ, Mrass P, Ritchie W, Doi A, Cavanagh LL, Tomura M, Sakaue-Sawano A, Kanagawa O, Miyawaki A, Hodgkin PD, Weninger W.
  • Journal Title: Nat Commun. Volume: 6 Issue: 1 Pages: 6301-6301
  • DOI: 10.1038/ncomms7301
  • Peer Reviewed / Open Access
• [Journal Article] Dermal Vγ4(+) γδ T Cells Possess a Migratory Potency to the Draining Lymph Nodes and Modulate CD8(+) T-Cell Activity through TNF-α Production. (2015)
  • Author(s): Nakamizo S, Egawa G, Tomura M, Sakai S, Tsuchiya S, Kitoh A, Honda T, Otsuka A, Nakajima S, Dainichi T, Tanizaki H, Mitsuyama M, Sugimoto Y, Kawai K, Yoshikai Y, Miyachi Y, Kabashima K.
  • Journal Title: Journal of Investigative Dermatology Volume: 135 Issue: 4 Pages: 1007-1015
  • DOI: 10.1038/jid.2014.516
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Cytotoxic T lymphocytes block tumor growth both by lytic activity and IFN-γ-dependent cell cycle arrest. (2015)
  • Author(s): Matsushita H, Hosoi A, Ueha S, Abe J, Fujieda N, Tomura M, Maekawa R, Matsushima K, Ohara O, Kakimi K.
  • Journal Title: Cancer Immunol Res. Volume: 3 Issue: 1 Pages: 26-36
  • DOI: 10.1158/2326-6066.cir-14-0098
  • Peer Reviewed
• [Journal Article] Tracking and quantification of dendritic cell migration and antigen trafficking between the skin and lymph nodes. (2014)
  • Author(s): Tomura M, Hata A, Matsuoka S, Shand FH, Nakanishi Y, Ikebuchi R, Ueha S, Tsutsui H, Inaba K, Matsushima K, Miyawaki A, Kabashima K, Watanabe T, Kanagawa O.
  • Journal Title: Sci Rep. Volume: 4 Issue: 1 Pages: 6030-6030
  • DOI: 10.1038/srep06030
  • NAID: 120005749847
  • Peer Reviewed / Open Access
• [Journal Article] Dental pulp dendritic cells migrate to regional lymph nodes (2014)
  • Author(s): Bhingare AC, Ohno T, Tomura M, Zhang C, Aramaki O, Otsuki M, Tagami J, Azuma M
  • Journal Title: J Dental Res Volume: 93 Issue: 3 Pages: 288-293
  • DOI: 10.1177/0022034513518223
  • Peer Reviewed
• [Journal Article] Tracking of intertissue migration reveals the origins of tumor-infiltrating monocytes. (2014)
  • Author(s): Shand FH, Ueha S, Otsuji M, Koid SS, Shichino S, Tsukui T, Kosugi-Kanaya M, Abe J, Tomura M, Ziogas J, Matsushima K.
  • Journal Title: Proc Natl Acad Sci USA. Volume: 111 Issue: 21 Pages: 7771-7776
  • DOI: 10.1073/pnas.1402914111
  • Peer Reviewed / Open Access
• [Journal Article] Contrasting Quiescent G0 Phase with Mitotic Cell Cycling in the Mouse Immune System. (2013)
  • Author(s): Tomura M, Sakaue-Sawano A, Mori Y, Takase-Utsugi M, Hata A, Ohtawa K, Kanagawa O, Miyawaki A.
  • Journal Title: Plos One Volume: 8 Issue: 9 Pages: e73801-e73801
  • DOI: 10.1371/journal.pone.0073801
  • Peer Reviewed
• [Journal Article] Qualitative rather than quantitative changes are hallmarks of fibroblasts in bleomycin-induced pulmonary fibrosis. (2013)
  • Author(s): Tsukui T, Ueha S, Abe J, Hashimoto S, Shichino S, Shimaoka T, Shand FH, Arakawa Y, Oshima K, Hattori M, Inagaki Y, Tomura M, Matsushima K.
  • Journal Title: Am J Pathol Volume: 183 Pages: 758-773
  • Peer Reviewed
• [Journal Article] Analysis of cell movement between skin and other anatomical sites in vivo using photoconvertible fluorescent protein "Kaede"-transgenic mice (2013)
  • Author(s): Tomura M and Kabashima K
  • Journal Title: Methods Mol Biol Volume: 961 Pages: 279-286
  • DOI: 10.1007/978-1-62703-227-8_18
  • ISBN: 9781627032261, 9781627032278
  • Peer Reviewed
• [Journal Article] Understanding of immune system by visualization of spatiotemporal regulation of immune cells in the entire body]. (2013)
  • Author(s): Tomura M.
  • Journal Title: Yakugaku Zasshi Volume: 133 Pages: 427-433
  • NAID: 130003361934
• [Presentation] Visualization of trafficking and presentation of tumor antigen by dendritic cells (2015)
  • Author(s): Michio Tomura, Ryoyo Ikebuchi
  • Organizer: Multi-dimensional fluorescence live imaging of cellular function and molecular activities (MDFLI
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 2015-01-26 – 2015-01-28
  • Invited
• [Presentation] 腸管免疫系における臓器間免疫細胞動態 (2014)
  • Author(s): 戸村道夫
  • Organizer: 第38回阿蘇シンポジウム
  • Place of Presentation: 熊本
  • Year and Date: 2014-07-25 – 2014-07-26
  • Invited
• [Presentation] 皮膚免疫制御機構における皮膚由来制御性T細胞の役割 (2014)
  • Author(s): 戸村道夫
  • Organizer: 第38回日本リンパ学会総会
  • Place of Presentation: 東京
  • Year and Date: 2014-06-20 – 2014-06-22
  • Invited
• [Presentation] Drastic increase of DC movement from immunizesd site and rapid replacement of antigen-carrying DC was induced in the draining lymph node during initiation of T cell proliferation (2013)
  • Author(s): Michio Tomura, Asako Sakaue-Sawano, Atushi Miyawaki, Osami Kanagawa
  • Organizer: 100th Annual meeting of the American association of immunologist
  • Place of Presentation: Honolulu, USA
• [Presentation] Simultaneous analysis of multiple fluorescent proteins and fluorochromes by a novel spectral flow cytometer (2013)
  • Author(s): Michio Tomura, Koji Futamura, Motohiro Furuki
  • Organizer: 100th Annual meeting of the American association of immunologist
  • Place of Presentation: Honolulu, USA
• [Presentation] Visualization of skin-derived DC dynamics by photoconvertible protein "KikGR" mice (2013)
  • Author(s): Michio Tomura, Akihiro Hata, Kenji Kabashima, Osami Kanagawa
  • Organizer: 23th日本樹状細胞研究会
  • Place of Presentation: 京都
• [Presentation] Characterization of effector and memory CD8+T cells migrated from virus infected skin (2013)
  • Author(s): Akihiro Hata, Michio Tomura
  • Organizer: KTCC 2013 International Workshop on T Lymphocytes
  • Place of Presentation: 京都
• [Presentation] フローサイトメーター及び生体内二光子レーザー顕微鏡による全身免疫細胞動態の可視化 (2013)
  • Author(s): 戸村 道夫
  • Organizer: 130622 第23回サイトメトリー学会 招待講演)
  • Place of Presentation: 東京
  • Invited
• [Presentation] Visualization of immune response by obserbing cell-cycle, movement and death. (2013)
  • Author(s): Michio Tomura
  • Organizer: COLD SPRING HARBOR ASIA CSHAsia 2013 Conference on New Advances in Optical Imaging of Live Cells and Organisms
  • Place of Presentation: 蘇州、中国
  • Invited
• [Presentation] Contrasting quiescent G0 phase with mitotic cell cycling in the mouse immune system (2013)
  • Author(s): Michio Tomura, Asako Sakaue-Sawano, Yoshiko Mori, Mitsuyo Takase, Akihiro Hata, Kenji Ohtawa, Osami Kanagawa, Atsushi Miyawaki
  • Organizer: 15th International Congress of Immunology
  • Place of Presentation: Milano, Italy
• [Presentation] Simultaneous analysis of multiple fluorescent proteins and fluorochromes by a novel spectral flow cytometer (2013)
  • Author(s): Michio Tomura, Koji Futamura, Motohiro Furuki
  • Organizer: 15th International Congress of Immunology
  • Place of Presentation: Milano, Italy
• [Presentation] Elicitation of a large number of DC migration and their rapid replacement in the draining LN during immune response (2013)
  • Author(s): Michio Tomura, Osami Kanagawa
  • Organizer: 4th Synthetic Immunology Workshop
  • Place of Presentation: 京都
• [Presentation] Contrasting quiescent G0 phase with mitotic cell cycling in the mouse immune system (2013)
  • Author(s): Michio Tomura, Asako Sakaue-Sawano, Yoshiko Mori, Mitsuyo Takase, Akihiro Hata, Kenji Ohtawa, Osami Kanagawa, Atsushi Miyawaki
  • Organizer: 第42回日本免疫学会学術集会
  • Place of Presentation: 千葉