Identification of antigens of tumor-infiltrated regulatory T cells and analysis of tumor suppression systems based on the antigens

• Project/Area Number: 18K07321
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 50020:Tumor diagnostics and therapeutics-related
• Research Institution: University of Toyama
• Principal Investigator: Kobayashi Eiji 富山大学, 学術研究部医学系, 助教 (70459733)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: T細胞受容体 / 制御性T細胞 / がん免疫 / Treg / TCR / 抗原 / 抗原同定

Outline of Final Research Achievements

In this research, we aimed to perform T cell receptor (TCR) repertoire analysis of tumor-infiltrating regulatory T cells (Treg) and determine antigens of the clonally expanded Treg cells using the TCR cloning systems that we had developed. To this end, we developed tumor-bearing mouse using B16F10 cells, prepared tumor infiltrating lymphocyte and analyzed the TCR repertoire. As a result, we determined TCRs that reacted to B16F10 cells. In addition, we have developed an antigen-determination system of CD8+ T cells which enabled us to determine antigens of CD8+ T cells. We will apply the system to determine antigens of the TCRs that we have cloned.

Academic Significance and Societal Importance of the Research Achievements

近年、免疫応答負に制御する制御性T細胞（Treg）の抑制を解除することで、がんに対する免疫応答を強化する新たな治療法の開発が注目されている。しかし、がん組織内でTregがどのような抗原を認識し、抗原特異的にがん免疫応答を抑制しているかに関しては、いまだ不明な点が多い。そこで、がん組織内でクローナルに集積するTregのT細胞受容体（TCR）の網羅的な解析を行い、それらTregの抗原を同定する。さらに、同定した抗原を用いて、Tregが抗原特異的にがん免疫応答を抑制しているか否かを検証する。本研究成果はTregを標的にした新たながん免疫療法に関する重要な知見を与えると考えられる。

Research Products

• [Journal Article] Analysis of TCR repertoire and PD-1 expression in decidual and peripheral CD8+ T cells reveals distinct immune mechanisms in miscarriage and preeclampsia. (2020)
  • Author(s): Morita K, Tsuda S, Kobayashi E, Hamana H, Tsuda K, Shima T, Nakashima A, Ushijima A, Kishi H, Saito S.
  • Journal Title: Front Immunol. Volume: 11 Pages: 1082-1082
  • DOI: 10.3389/fimmu.2020.01082
  • Peer Reviewed / Open Access
• [Journal Article] Relationship between T cell receptor clonotype and PD-1 expression of tumor-infiltrating lymphocytes in colorectal cancer. (2020)
  • Author(s): Sukegawa K, Shitaoka K, Hamana H, Kobayashi E, Miyahara Y, Fujii K, Tsuda K, Saeki S, Nagata T, Ozawa T, Saito S, Fujii T, Muraguchi A, Shiku H, Kishi H.
  • Journal Title: Eur. J. Immunol. Volume: 50 Issue: 10 Pages: 1580-1590
  • DOI: 10.1002/eji.201948399
  • Peer Reviewed
• [Presentation] 大腸がん浸潤リンパ球におけるTCRクロノタイプとPD-1の発現 (2020)
  • Author(s): 岸 裕幸，下岡清美，小林栄治，宮原慶裕，津田 桂，佐伯しおり，長田拓哉，村口 篤，珠玖 洋
  • Organizer: 第79回日本癌学会学術総会
• [Presentation] Development of a novel tumor antigen-specific TCR cloning system using a microarray chip (2020)
  • Author(s): KOBAYASHI Eiji, OZAWA Tatsuhiko, HAMANA hiroshi, MURAGUCHI Atsushi, KISHI Hiroyuki
  • Organizer: 第24回日本がん免疫学会総会
• [Presentation] ISAAC法を用いたTCR様抗体の迅速作製とその応用 (2020)
  • Author(s): 小澤龍彦，小林栄治，浜名 洋，村口 篤，岸 裕幸
  • Organizer: 第24回日本がん免疫学会総会
• [Presentation] Jurkat細胞を用いたTCRのネオアンチゲン反応性解析 (2020)
  • Author(s): 小澤龍彦，小林栄治，浜名 洋，村口 篤，岸 裕幸
  • Organizer: 第28回日本胎盤学会学術集会
• [Presentation] 正常妊娠と異常妊娠における脱落膜CD8+T細胞のTCRレパートリーとPD-1の発現についての検討 (2020)
  • Author(s): 森田恵子，津田さやか，小林栄治，浜名 洋，津田 桂，島 友子，中島彰俊，岸 裕幸，齋藤 滋
  • Organizer: 第28回日本胎盤学会学術集会
• [Presentation] A novel TCR cloning system of peptide-specific T cells using immunospot array assay on a chip(T-ISAAC) technology (2019)
  • Author(s): Kobayashi E, Ozawa T, Hamana H, Shitaoka K, Muraguchi A, Kishi H.
  • Organizer: International Conference on Lymphocyte Engineering 2019
  • Int'l Joint Research
• [Presentation] 患者HLA遺伝子導入乳がん細胞株を用いた乳がん患者TIL中の腫瘍反応性TCRの探索 (2019)
  • Author(s): 浜名 洋，下岡 清美，祐川 健太，佐伯しおり，長田任一哉，小林 栄治，小澤 龍彦，藤井 努，村口 篤，岸 裕幸
  • Organizer: 第23回日本がん免疫学会総会
• [Presentation] 腫瘍浸潤CD8+ T細胞におけるPD-1の発現はTCRレパトアに規定される (2019)
  • Author(s): 祐川 健太，佐伯しおり，下岡 清美，浜名 洋，宮原 慶裕，小林 栄治，長田任一哉，小澤 龍彦，藤井 努，珠玖 洋，村口 篤，岸 裕幸
  • Organizer: 第23回日本がん免疫学会総会
• [Presentation] Clonally expanded populations of cytotoxic T cell in TILs and peripheral blood in endometrial cancer patients (2019)
  • Author(s): 津田 桂，浜名 洋，中島 彰俊，森田 恵子，津田さやか，島 友子，下岡 清美，小林 栄治，小澤 龍彦，村口 篤，岸 裕幸
  • Organizer: 第23回日本がん免疫学会総会
• [Presentation] 腫瘍浸潤CD8+ T細胞のTCRとPD-1の発現との関連：大腸癌における考察． (2019)
  • Author(s): 岸 裕幸，祐川 健太，下岡 清美，浜名 洋，小林 栄治，津田 桂，長田任一哉，佐伯しおり，小澤 龍彦，齋藤 滋，藤井 努，村口 篤
  • Organizer: 第78回日本癌学会学術総会
• [Presentation] Screening of tumor-reactive TCRs from TILs of breast cancer patients using a patients' HLA-transduced breast cancer cell line. (2019)
  • Author(s): Hamana H, Shitaoka K, Sukegawa K, Saeki S, Nagata T, Kobayashi E, Ozawa T, Fujii T, Muraguchi A, Kishi H
  • Organizer: 第49回日本免疫学会学術集会
• [Presentation] Common TCRβ repertoire was found in both decidual and peripheral CD8+ T cells in normal term pregnancy (2019)
  • Author(s): Morita K, Kobayashi E, Tsuda S, Shitaoka K, Ozawa T, Hamana H, Saito S, Kishi H
  • Organizer: 第49回日本免疫学会学術集会
• [Presentation] Cloning of tumor antigen-specific TCRs using immunospot array assay on a chip (T-ISAAC) technology (2019)
  • Author(s): Kobayashi E, Ozawa T, Hamana H, Shitaoka K, Muraguchi A, Kishi H
  • Organizer: 第49回日本免疫学会学術集会
• [Presentation] TCR repertoire analysis of cytotoxic T cells in tumor-infiltrated lymphocytes and peripheral blood lymphocytes in endometrial cancers (2019)
  • Author(s): Tsuda K, Hamana H, Nakashima A, Tsuda S, Shima T, Shitaoka K, Kobayashi E, Ozawa T, Kishi H
  • Organizer: 第49回日本免疫学会学術集会
• [Presentation] TCR repertoire analysis of peptide-specific T cells using immunospot array assay on a chip (T-ISAAC) technology (2018)
  • Author(s): KOBAYASHI Eiji, MURAGUCHI Atsushi, KISHI Hiroyuki
  • Organizer: 第77回日本癌学会学術総会
• [Presentation] TCR repertoire analysis of peptide-specific T cells using immunospot array assay on a chip (T-ISAAC) technology (2018)
  • Author(s): KOBAYASHI Eiji, OZAWA Tatsuhiko, HAMANA Hiroshi, SHITAOKA Kiyomi, MURAGUCHI Atsushi and KISHI Hiroyuki
  • Organizer: 第47回日本免疫学会学術集会