Generation of antigen presenting cells applicable to standardized evaluation of immune status following immunotherapy targeting minor antigens.

• Project/Area Number: 24591435
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Hematology
• Research Institution: Aichi Cancer Center Research Institute
• Principal Investigator: AKATSUKA Yoshiki 愛知県がんセンター(研究所), 腫瘍免疫学部, 客員研究員 (70333391)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2014: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 抗原提示細胞 / マイナー組織適合抗原 / 同種造血細胞移植 / 同種移植 / 免疫学 / 細胞治療 / ワクチン

Outline of Final Research Achievements

We sought to generate antigen presenting cells (APCs) that can be used for standardization when assessing immune responses following immunotherapies. HLA-deficient K562 was mainly tested using various types of approaches to control antigen expression level. Antigenic peptides were co-expressed with GFP or NGFR to estimate antigen expression levels indirectly. Because modification of Kozak sequence was incapable for the purpose, tetracycline-inducible system was incorporated. The latter could control cell surface GFP/NGFR expression level in a wide range from null to around several hundred times when assessed by flow cytometry, however the cognate CTLs was sensitive enough to recognize target cells without any GFP/NGFR expression. To this end, we are currently modifying the APC system so as to control the cell surface HLA expression level, which will be suitable APCs expressing natural level of antigenic peptide internally for testing clinical samples.

Research Products

• [Journal Article] Induction of HLA-B*40:02-restricted T cells possessing cytotoxic and suppressive functions against haematopoietic progenitor cells from a patient with severe aplastic anaemia. (2015)
  • Author(s): Inaguma Y, Akatsuka Y, Hosokawa K, Maruyama H, Okamoto A, Katagiri T, Shiraishi K, Murayama Y, Tsuzuki-Iba S, Mizutani Y, Nishii C, Yamamoto N, Demachi-Okamura A, Kuzushima K, Ogawa S, Emi N, Nakao S.
  • Journal Title: Br J Haematol. Volume: in press Issue: 1 Pages: 131-134
  • DOI: 10.1111/bjh.13464
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H. (2014)
  • Author(s): Inaguma, Y., Akahori, Y., Murayama,Y., Shiraishi, K., Tsuzuki-Iba, S., Endoh, A., Tsujikawa, J., Demachi-Okamura, A., Hiramatsu, K., Saji, H., Yamamoto, Y., Yamamoto, N., Nishimura, Y., Takahashi, T., Kuzushima, K., Emi, N., and Akatsuka, Y.
  • Journal Title: Gene Therapy Volume: 3 Issue: 6 Pages: 575-584
  • DOI: 10.1038/gt.2014.30
  • Peer Reviewed
• [Journal Article] Adoptive transfer of genetically engineered WT1-specific cytotoxic T lymphocytes does not induce renal injury (2014)
  • Author(s): Asai H, Fujiwara H, Kitazawa S, Kobayashi N, Ochi T, Miyazaki Y, Ochi F, Akatsuka Y, Okamoto S, Mineno J, Kuzushima K, Ikeda H, Shiku H, Yasukawa M
  • Journal Title: J Hematol Oncol. Volume: 7 Issue: 1 Pages: 3-3
  • DOI: 10.1186/1756-8722-7-3
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] An HLA-modified ovarian cancer cell line induced CTL responses specific to an epitope derived from claudin-1 presented by HLA-A*24:02 molecules (2013)
  • Author(s): Kondo S, Demachi-Okamura A, Hirosawa T, Maki H, Fujita M, Uemura Y, Akatsuka Y, Yamamoto E, Shibata K, Ino K, Kikkawa F, Kuzushima K.
  • Journal Title: Hum Immunol Volume: 74 Issue: 9 Pages: 1103-1110
  • DOI: 10.1016/j.humimm.2013.06.030
  • Peer Reviewed
• [Journal Article] Recognition of a natural WT1 epitope by a modified WT1 peptide-specific T-cell receptor. (2012)
  • Author(s): Tamanaka T, et al.
  • Journal Title: Anticancer Res. Volume: 32 Pages: 5201-5299
  • Peer Reviewed
• [Presentation] 親和性の異なるHLA-A2拘束性HA-1H特異的CAR-T細胞の機能解析. (2014)
  • Author(s): 赤塚美樹，赤堀泰，稲熊容子，葛島清隆，恵美宣彦
  • Organizer: 第6回血液疾患免疫療法研究会学術集会
  • Place of Presentation: 芝蘭会館 山内ホール(京都市）
  • Year and Date: 2014-09-06
• [Presentation] 異なった親和性をもつHLA-A2拘束性マイナー抗原HA-1Hを特異的CAR-T細胞の機能解析． (2014)
  • Author(s): 赤塚美樹、赤堀泰、稲熊容子、西村泰治、岡村文子、葛島清隆、恵美宣彦
  • Organizer: 第18回日本がん免疫学会総会
  • Place of Presentation: ひめぎんホール（松山市）
  • Year and Date: 2014-07-31
• [Presentation] HLA-A2拘束性に提示されたマイナー抗原HA-1Hを認識する抗体の単離とCAR-Tの機能 (2013)
  • Author(s): 赤塚美樹、赤堀泰、稲熊容子、ら
  • Organizer: 第17回日本がん免疫学会総会
  • Place of Presentation: ANAクラウンプラザホテル宇部（宇部市）
• [Presentation] Construction and molecular characterization of a single chain antibody specific for HA-1 minor H antigen. (2013)
  • Author(s): Akatsuka A, Demachi-Okamura A, Yamamoto Y, ら
  • Organizer: 第72回日本癌学会総会
  • Place of Presentation: 横浜・パシフィコ横浜（横浜市）
• [Presentation] Construction and Molecular Characterization Of a T-Cell Receptor-Like Antibody and CAR-T Cells Specific For Minor Histocompatibility Antigen HA-1H. (2013)
  • Author(s): Inaguma Y, Akahori Y, Akatsuka Y, et al.
  • Organizer: The 55th ASH Annual Meeting,
  • Place of Presentation: National Cancer Institute （New Orleans市, 米国）
• [Presentation] Vaccination With Minor Histocompatibility Antigen-Derived Peptides In Post-Transplant Patients With Hematological Malignancies - Preliminary Results. (2012)
  • Author(s): Yoshiki Akatsuka, et al.
  • Organizer: 2nd International Workshop on the Biology, Prevention, and Treatment of Relapse After Hematopoietic Stem Cell Transplantation.
  • Place of Presentation: Natcher Auditorium ( Bethesda, MD, USA)
• [Presentation] HLA-A2拘束性に提示されたマイナー抗原HA-1Hペプチドを認識する抗体の単離とその臨床応用に向けての検討
  • Author(s): 赤堀 泰、ら
  • Organizer: 第35回日本造血細胞移植学会
  • Place of Presentation: 石川県立音楽堂(金沢)