マウスTL抗原を標的とした免疫療法モデルの開発

• Project/Area Number: 12217170
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Aichi Cancer Center Research Institute
• Principal Investigator: 高橋 利忠 愛知県がんセンター(研究所), 研究所長 (00124529)
• Co-Investigator(Kenkyū-buntansha): 辻村 邦夫 愛知県がんセンター(研究所), 腫瘍免疫学部, 室長 (10227407) ; 赤塚 美樹 愛知県がんセンター(研究所), 腫瘍免疫学部, 室長 (70333391) ; 中山 俊憲 千葉大学, 大学院・医学研究科, 助教授 (50237468)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥66,200,000 (Direct Cost: ¥66,200,000); Fiscal Year 2004: ¥12,000,000 (Direct Cost: ¥12,000,000); Fiscal Year 2003: ¥12,000,000 (Direct Cost: ¥12,000,000); Fiscal Year 2002: ¥10,000,000 (Direct Cost: ¥10,000,000); Fiscal Year 2001: ¥12,200,000 (Direct Cost: ¥12,200,000); Fiscal Year 2000: ¥20,000,000 (Direct Cost: ¥20,000,000)
• Keywords: マウスTI抗原 / 細胞傷害性T細胞 / テトラマー / 杭体活性 / T細胞レセプターキメラ分子 / 抗体キメラ分子 / 免疫監視機構 / NBU化学発がん剤 / マウスTL抗原 / 樹状細胞 / B細胞 / CD8 / T細胞レセプター / 腸上皮間リンパ球 / 胸腺細胞 / 細胞傷害性T細胞前駆細胞 / 抗原物質非依存性エピトープ / 主要組織適合抗原 / NKT細胞 / TLテトラマー / 免疫モニタリング / 腫瘍免疫療法

Research Abstract

(1)組み換え技術を用いて、TL抗原特異的エフェクターT細胞株を樹立すべく、TL特異的細胞傷害性T細胞(CTL)クローンからT細胞レセプター(TCR)遺伝子を、また抗TL抗体産生ハイブリドーマから抗体遺伝子の単離を行った。単離したTCRおよび抗体遺伝子が単鎖構造を取るような組み換え遺伝子を構築し、さらにシグナル伝達分子であるCD3ζ遺伝子の細胞内ドメインとタンデムに結合させたキメラ遺伝子を作成した。このうち、抗体キメラ分子については、活性化T細胞表面に発現できることを確認した。
(2)C3HとB6マウスに化学発がん剤NBUを経口投与すると、C3HにおけるTL陽性腫瘍の発症時期がB6に比べて遅延した(統計学的には有為差は無い)。また、C3Hが同系のTL遺伝子導入マウス由来のTL陽性皮膚を拒絶するのに対し、B6では拒絶が見られなかった。In vivo抗TL活性の異なるC3HとB6を解析し、H-2K^b分子の発現により、TL特異的CTLの不活化が起こることを示唆した。また、TLテトラマー陽性細胞率および血清中の抗TL抗体活性はB6よりC3Hにおいて高く、in vivoの抗TL活性との相関が見られたが、(B6×C3H)F_1マウスをrecipientとして用いた各個体レベルでの検索では、これらin vitroの成績とin vivo抗TL活性とには明らかな一致が見られなかった。

Research Products

• [Journal Article] Retroviral vector backbone immunogenicity : identification of cytotoxic T cell epitopes in retroviral vector-packaging sequences. (2005)
  • Author(s): Kondo, E.
  • Journal Title: Gene Ther. 12 Pages: 252-258
• [Journal Article] Thymus-leukemia antigen (TL) as a major histocompatibility complex (MHC) class Ib molecule and tumor-specific antigen. (2004)
  • Author(s): Tsujimura, K.
  • Journal Title: Cancer Sci. 95 Pages: 469-474
  • NAID: 10013723319
• [Journal Article] Immunity against mouse thymus-leukemia antigen (TL) protects against development of T lymphomas induced by a chemical carcinogenic agent, N-butyl-N-nitrosourea. (2004)
  • Author(s): Tsujimura, K.
  • Journal Title: Cancer Sci. 95 Pages: 914-919
• [Journal Article] Identification of novel CTL epitopes of CMV-pp65 presented by avariety of HLA alleles. (2004)
  • Author(s): Rondo, E.
  • Journal Title: Blood 103 Pages: 630-638
• [Journal Article] Interferon-γ differentially regulates susceptibility of lung cancer cells to telomerase-specific cytotoxic T lymphocytes. (2004)
  • Author(s): Tajima, K.
  • Journal Title: Int.J.Cancer 110 Pages: 403-412
• [Journal Article] Human TMSB4Y gene encodes a novel HLA-A^*3303 -restricted minor histocompatibility antigen in its 5' untranslated region. (2004)
  • Author(s): Torikai, H.
  • Journal Title: J.Immunol. 173 Pages: 7046-7054
• [Publications] Tsujimura, K.: "Thymus antigen (TL)-specific CTL recognize a1/a2 domain of TL free from antigenic peptides."Int.Immunol.. 15. 1319-1326 (2003)
• [Publications] Akatsuka, Y.: "Identification of a polymorphic gene, BCL2A1, encoding two novel hematopoietic lineage specific minor histocompatibility antigens."J.Exp.Med.. 197. 1489-1500 (2003)
• [Publications] Miyazaki, M.: "Potential limitations in using minor histocompatibility antigen-specific cytotoxic T cells for targeting solid tumor cells."Clin.Immunol.. 107. 198-201 (2003)
• [Publications] Kondo, E: "Identification of novel CTL epitopes of CMV-pp65 presented by a variety of HLA alleles."Blood. 103. 630-638 (2004)
• [Publications] Tajima, K.: "Interferon-γ differentially regulates susceptibility of lung cancer cells to telomerase-specific cytotoxic T lymphocytes."Int.J.Cancer. (in press). (2004)
• [Publications] Tsujimura, K.: "Thymus-leukemia (TL) antigen as a major histocompatibility complex (MHC) class Ib molecule and tumor specific antigen."Cancer Sci.. (in press). (2004)
• [Publications] Yazaki, M.: "Generation of HLA-Cw specific cytotoxic T-lymphocytes from cord blood used for cord blood stem cell transplantation"Brit. J. Haematol.. 117. 893-898 (2002)
• [Publications] Akatsuka, Y.: "Efficient cloning and expression of HLA class I cDNA in human B-lymphoblastoid cell lines"Tissue Antigens. 59. 502-511 (2002)
• [Publications] Kondo, E: "Efficient generation of antigen-specific cytotoxic T cells using retrovirally transduced CD40-activated B cells"J. Immunol.. 169. 2164 (2002)
• [Publications] Mutimer, H.P.: "Association between immune recovery uveitis and a diverse intraocularcytomegalovirus-specific cytotoxic T cell response"J. Infect. Dis.. 186. 701-705 (2002)
• [Publications] Akatsuka, Y.: "Targeted cloning of cytotoxic T cells specific for minor histocom patibility antigens restricted by HLA class I molecules of interest"Transplantation. 74. 1773-1780 (2002)
• [Publications] Kuzushima, K.: "Tetramer-assisted identification and characterization of epitopes recognized by HLA A^*2402-restricted Epstein-Barr virus specific CD8^+ T cells"Blood. 101. 1460-1468 (2002)
• [Publications] Tsujimura, K.: "The binding of thymus leukemia(TL) antigen tetramers to normal intestinal intraepithelial lymphocytes and thymocytes"J.Immunol.. 167. 759-764 (2001)
• [Publications] Aoki, H.: "Dendritic cells pulsed with tumor extract-cationic liposome complex increase the induction of cytotoxic T lymphocytes in mouse brain tumor"Cancer Immunol. Immunother.. 50. 463-468 (2001)
• [Publications] Pei, J.: "Generation of HLA-C-specific cytotoxic T cells in association with marrow graft rejection : analysis of alloimmunity by T-cell cloning and testing of T-cell-receptor rearrangements"Biol Blood Marrow Transplant.. 7. 378-383 (2001)
• [Publications] Akatsuka, Y.: "Determination of intronic sequences adjacent to an exon using polymerase chain reaction and genomic DNA library constructed by TA cloning"Anal Biochem.. 289. 289-292 (2001)
• [Publications] Brickner, A.G.: "The immunogenicity of a new human minor histocompatibility antigen results from differential antigen processing"J. Exp. Med.. 193. 195-206 (2001)
• [Publications] Yazaki, M.: "Generation of HLA-Cw specific cytotoxic T-lymphocytes from cord blood used for cord blood stem cell transplantation"Brit. J. Haematol.. (In press).
• [Publications] Iwase,S.: "Comparison of anti-tumor responses against TL positive lymphoma induced by skin grafting and dendritic cell immunization"Microbiol.Immunol.. 44. 609-618 (2000)
• [Publications] Tsujimura,K.: "The epitope detected by CTL against thymus leukemia (TL) antigen is TAP-independent"Int.Immunol.. 12. 1183-1191 (2000)
• [Publications] Yazaki,M.: "Generation of HLA-A2 subtype specific T lymphocytes from cord blood used for cord blood stem cell transplantation"Bone Marrow Transplant.. 26. 451-454 (2000)
• [Publications] Matsuda,J.L.: "Tracking the response of NKT cells to a glycolipid antigen using CD1d tetramers : The response to α-galact osylceramide is dynamic and highly compartmentalized"J.Exp.Med.. 192. 741-754 (2000)
• [Publications] Yamashita,M.: "TCR-induced calcineurin activation regulates Th2 cell development by modifying the IL-4 receptor signaling complex"J.Exp.Med.. 191. 1869-1879 (2000)
• [Publications] Ikehara,Y.: "CD4+ Vα 14NKT cells are essential for acceptable of intrahepatic rat islet xenografts in mice treated with anti-CD4 antibody"J.Clin.Invest.. 105. 1761-1767 (2000)