In vivo analysis of immune response by a newly developed 4-color immunohistological staining method: Rejection or tolerance following rat liver transplantation

• Project/Area Number: 17K08518
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General anatomy (including histology/embryology)
• Research Institution: Dokkyo Medical University
• Principal Investigator: Matsuno Kenjiro 獨協医科大学, 医学部, 特任教授 (20094047)
• Co-Investigator(Kenkyū-buntansha): 上田 祐司 獨協医科大学, 医学部, 准教授 (10364556)
• Project Period (FY): 2017-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: ラット肝移植 / 拒絶反応モデル / 免疫寛容モデル / 3重免疫酵素染色 / 4重免疫蛍光染色 / 樹状細胞 / 免疫組織学 / ドナー特異的抗体 / 樹状細胞亜群 / 移植・再生医療 / 免疫学 / 解剖学

Outline of Final Research Achievements

DST-antibodies and anti-donor MHCII antibodies could suppress the CD8+ T-cell-mediated liver transplant rejection by depleting donor immunogenic DCs, blocking the direct pathway of allorecognition. Donor MHCII-specific antibodies may be applicable as a selective suppressant of anti-donor immunity for clinical liver transplantation without the cellular damage of donor MHCII-negative graft cells and recipient cells.　Four color immunofluorescence staining confirmed the presence of donor MHCII+CD103+ DC subsets clustering with proliferating EdU+ cells, mostly recipient T cells, which suggests that the DC cluster is a site for the direct alloantigen presentation. DST pretreatment more strongly suppressed the intragraft response and the transplant rejection, which may be due to the presence of regulatory T cells. Additionally, allogeneic T cells may be clinically applicable as vaccine vectors for polytopical prophylactic antibody production.

Academic Significance and Societal Importance of the Research Achievements

学術的意義：4重免疫蛍光染色を確立し、肝移植拒絶群では、移植肝内のDC亜群が強いアロ抗原刺激能をもち拒絶反応を誘導すること、DST寛容群では、ドナーDC亜群を除去して拒絶反応を有意に抑制する抗ドナーMHCⅠ抗体が産生され、移植免疫応答を抑制する制御性T細胞も誘導されることを証明し、移植肝拒絶とDSTによる免疫寛容のメカニズムを明らかにした。
社会的意義：抗ドナーMHCⅡ抗体が肝移植の拒絶反応の選択的抑制剤として使えることと、ドナーT細胞が全身のリンパ節で多所性に中和抗体を誘導できるワクチンとして使用可能であることを特許出願し、基礎研究成果の臨床応用の道を開いた。

Research Products

• [Journal Article] Suppression of liver transplant rejection by anti-donor MHC antibodies via depletion of donor immunogenic dendritic cells (2020)
  • Author(s): Hisashi Ueta,Xue-Dong Xu,Bin Yu,Yusuke Kitazawa,Enqiao Yu,Yoshiaki Hara,Miwa Morita-Nakagawa,Shu Zhou,Yasushi Sawanobori,Satoshi Ueha,Kazuhito Rokutan,Toshiya Tanaka,Nobuko Tokuda,Kouji Matsushima,Kenjiro Matsuno
  • Journal Title: International Immunology Volume: 33 Pages: 261-272
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Novel Targeting to XCR1+ Dendritic Cells Using Allogeneic T Cells for Polytopical Antibody Responses in the Lymph Nodes (2019)
  • Author(s): Kitazawa Yusuke、Ueta Hisashi、Sawanobori Yasushi、Katakai Tomoya、Yoneyama Hiroyuki、Ueha Satoshi、Matsushima Kouji、Tokuda Nobuko、Matsuno Kenjiro
  • Journal Title: Frontiers in Immunology Volume: 10 Pages: 1195-1195
  • DOI: 10.3389/fimmu.2019.01195
  • Peer Reviewed / Open Access
• [Journal Article] Single blood transfusion induces the production of donor-specific alloantibodies and regulatory T cells mainly in the spleen. (2018)
  • Author(s): Ueta H, Kitazawa Y, Sawanobori Y, Ueno T, Ueha S, Matsushima K, Matsuno K.
  • Journal Title: Int Immunol Volume: 30 Issue: 2 Pages: 53-67
  • DOI: 10.1093/intimm/dxx078
  • Peer Reviewed / Open Access
• [Journal Article] Rapid immunosurveillance by recirculating lymphocytes in the rat intestine: critical role of unsulfated sialyl-Lewis X on high endothelial venules of the Peyer’s patches (2018)
  • Author(s): Uchida Tomomi、Ueta Hisashi、Xu Xue-Dong、Hirakawa Jotaro、Tahara Kazunori、Zhou Shu、Sawanobori Yasushi、Simmons Szandor、Kitazawa Yusuke、Kawashima Hiroto、Matsuno Kenjiro
  • Journal Title: International Immunology Volume: 30 Issue: 1 Pages: 23-33
  • DOI: 10.1093/intimm/dxx072
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Novel DC targeting by allogenic T-cells for multifocal prophylactic antibody productio (2019)
  • Author(s): Ueta H, Kitazawa Y, Sawanobori Y, Katakai T, Ueha S, Matsushima K, Tokuda N, Matsuno K
  • Organizer: 第48回日本免疫学会学術集会（ベストプレゼンテーション賞受賞）
• [Patent(Industrial Property Rights)] T細胞ワクチン (2019)
  • Inventor(s): 松野健二郎、上田祐司、北沢祐介
  • Industrial Property Rights Holder: 獨協医科大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2019-024219
  • Filing Date: 2019