Characterization of HTLV-1 specific CTLs detected in HAM patients

• Project/Area Number: 21K08494
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 54030:Infectious disease medicine-related
• Research Institution: Kumamoto University
• Principal Investigator: Sugata Kenji 熊本大学, ヒトレトロウイルス学共同研究センター, 講師 (10650616)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2023: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2022: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2021: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: HTLV-1 / HAM / HLA-A24 / Tax / 特異的CD8T細胞 / 抗原特異的T細胞 / HTLV1 / CTL

Outline of Research at the Start

これまでHAM患者での病態解析は末梢血T細胞を用いて行われてきたが、有用なHAMマーカーは見つかっていない。本研究ではHAM病態形成の場である脳脊髄と末梢血でのT細胞を１細胞解析を行うことで、HAM患者特有の遺伝子発現を同定する。また、それらの結果とTCR-peptide predictionアルゴリズム解析を組み合わせることで脳脊髄に浸潤したT細胞が認識する抗原を同定し、HAM患者でのHTLV-1抗原特異的CTLが特異的に発現する遺伝子（HAMマーカー）の探索も同様に行う。

Outline of Final Research Achievements

To understand pathogenesis of HTLV-1-associated inflammatory diseases, we performed integrated single cell analysis using in peripheral blood and cerebrospinal fluid (CSF) cells derived from HAM/TSP patients. Among various cell types, CD8 T cells were increased in the CSF and contained significantly expanded anti-Tax cytotoxic T-lymphocytes (CTLs). Transcriptome analysis demonstrated that anti-Tax CTLs in the CSF showed reduced activation whereas enhanced pro-apoptotic and exhausted phenotype. Furthermore, we found that HLA-A24 presents two different peptides of Tax depending on Tax genotypes. That affects results of HLA multiuser staining in specific CD8 T cells of the infected patients

Academic Significance and Societal Importance of the Research Achievements

本研究ではHAM患者の脊髄病変に浸潤しているHTLV-1抗原特異的CTLの遺伝子発現プロファイルに基づき、末梢血での同CTLが発現するHAMの発症リスク遺伝子を発見することを目的としている。HTLV-1特異的CTLが発現するHAMの発症リスク遺伝子を探索するため、次世代シークエンサーを用いて脳脊髄液中のCTLの1細胞遺伝子発現解析とTCRレパトア解析を行った。TCR-CDR3やclonotypeに基づき脳脊髄液中のHTLV-1特異的CTLとの比較を行うことでHAMの発症リスク遺伝子群の発見を目指し、それらを利用してHAMの早期発見・治療開始に繋げる。

Research Products

• [Journal Article] HIV-Tocky system to visualize proviral expression dynamics (2024)
  • Author(s): O, Reda., K, Monde., K, Sugata., A, Rahman., W, Sakhor., SA, Rajib., SN, Sithi., BJY, Tan., K, Niimura., C, Motozono., K, Maeda., M, Ono., H, Takeuchi and Y, Satou. 4.巻 7
  • Journal Title: Communications biology Volume: 7 Issue: 1 Pages: 334-350
  • DOI: 10.1038/s42003-024-06025-8
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Association of Clostridium butyricum Therapy Using the Live Bacterial Product CBM588 with the Survival of Patients with Lung Cancer Receiving Chemoimmunotherapy Combinations (2023)
  • Author(s): Tomiya Y, Sakata S, Imamura K, Iyama S, Jodai T, et al.
  • Journal Title: Cancers Volume: 16 Issue: 1 Pages: 47-47
  • DOI: 10.3390/cancers16010047
  • Peer Reviewed / Open Access
• [Journal Article] Immunopathology of HTLV-1-Associated Adult T-Cell Leukemia/Lymphoma: A Comprehensive Review. (2023)
  • Author(s): Nakahata S, Enriquez-Vera D, Jahan MI, Sugata K, Satou Y.
  • Journal Title: Biomolecules Volume: 13 Issue: 10 Pages: 1543-1543
  • DOI: 10.3390/biom13101543
  • Peer Reviewed / Open Access
• [Journal Article] HTLV-1 persistence and leukemogenesis: A game of hide-and-seek with the host immune system (2022)
  • Author(s): Tan Benjy J. Y.、Sugata Kenji、Ono Masahiro、Satou Yorifumi
  • Journal Title: Frontiers in Immunology Volume: 13 Pages: 01-12
  • DOI: 10.3389/fimmu.2022.991928
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] HTLV-1 infection promotes excessive T cell activation and transformation into adult T cell leukemia/lymphoma (2021)
  • Author(s): Tan Benjy J.Y.、Sugata Kenji et al.
  • Journal Title: Journal of Clinical Investigation Volume: 131 Issue: 24
  • DOI: 10.1172/jci150472
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] HLA-A24拘束性の異なるTaxエピトープを認識する特異的TCRの解析 (2023)
  • Author(s): 菅田謙治, Benjy Tan Jek Yang, 高鳥光徳, Md Belal Hossain, Rajib Md Samiul Alam, Omnia Reda, 徳永雅仁, 野村隼也, 増田曜章, 中島誠, 佐藤知雄, 植田光晴, 宇都宮與, 山野嘉久, 佐藤賢文
  • Organizer: 第9回 日本HTLV-1学会
• [Presentation] Single cell analysis of cerebrospinal cord fluid (CSF) revealed expansion of anti-viral CTLs in HTLV-1-associated myelopathy patients (2023)
  • Author(s): Kenji Sugata
  • Organizer: 2nd Oxford-Kumamoto U Joint Symposium
• [Presentation] HAM患者末梢血および脳脊髄液のシングルセル解析によるHAM病態解明と疾患サロゲートマーカーの探索研究 (2022)
  • Author(s): 菅田謙治
  • Organizer: 第8回日本HTLV-1学会
• [Presentation] Integrated single cell analysis of HTLV-1 specific CD8 T cells in peripheral blood and cerebrospinal fluid from HAM patients (2022)
  • Author(s): Kenji Sugata
  • Organizer: 23rd KUMAMOTO AIDS Seminar
• [Presentation] TCR配列に基づいた末梢血と脳脊髄液中のHTLV-1特異的CD8+T細胞の包括的解析 (2021)
  • Author(s): 菅田謙治
  • Organizer: 第7回HTLV-1学会
• [Book] HTLV-1による成人T細胞白血病・リンパ腫の発症機構 (2023)
  • Author(s): 菅田謙治, 佐藤賢文
  • Total Pages: 6
  • Publisher: 日本臨牀 81 (増刊号3)
• [Patent(Industrial Property Rights)] 抗原ペプチド、核酸、ベクター、HLA／抗原ペプチド複合体、検出試薬、医薬組成物 (2023)
  • Inventor(s): 佐藤賢文、菅田謙治
  • Industrial Property Rights Holder: 佐藤賢文、菅田謙治
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2023-178883
  • Filing Date: 2023