白血病を標的とした新たな免疫細胞治療法の開発研究

• Project/Area Number: 13218021
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokyo
• Principal Investigator: 千葉 滋 東京大学, 医学部附属病院, 助教授 (60212049)
• Co-Investigator(Kenkyū-buntansha): 鈴木 隆浩 東京大学, 医学部・附属病院, 医員 ; 熊野 恵城 東京大学, 医学部・附属病院, 医員
• Project Period (FY): 2001 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥37,300,000 (Direct Cost: ¥37,300,000); Fiscal Year 2004: ¥10,000,000 (Direct Cost: ¥10,000,000); Fiscal Year 2003: ¥9,800,000 (Direct Cost: ¥9,800,000); Fiscal Year 2002: ¥9,000,000 (Direct Cost: ¥9,000,000); Fiscal Year 2001: ¥8,500,000 (Direct Cost: ¥8,500,000)
• Keywords: 白血病 / 造血器腫瘍 / 同種移植 / HLA / 造血幹細胞 / T前駆細胞 / Notch / Delta1 / 抗CD52モノクローナル抗体 / HLA不一致同種造血幹細胞移植 / NKT細胞 / 免疫 / 細胞療法 / ミニ移植 / CD52 / 慢性骨髄性白血病 / 融合遺伝子 / BCR / ABL / 腫瘍特異的抗原 / 樹状細胞

Research Abstract

白血病を中心とした造血器腫瘍を標的とし、HLA2座以上不一致の血縁者をドナーとして行う移植に関し、方法論改善に向けた開発的研究を行った。HLA2座以上不一致ドナーからの移植では、致死的な移植片対宿主病(GVHD)を回避するため、何らかの方法でT細胞を除去または強力に抑制することが必要である。我々は、抗CD52モノクローナル抗体を用いたin vivo T細胞除去法により臨床試験を行い、HLA2座以上不一致移植においても重症GVHDが回避可能であることを見出した。しかしこの方法では、免疫再構築の遅れが顕著であった。動物実験では、GVHDモデル系において、ドナーT細胞を除去してもT前駆細胞を造血幹細胞と共に輸注することにより重症GVHDを回避しつつ早期免疫再構築を図ることが明らかにされている。この原理を臨床に応用することに向けて研究を行い、以下の成果を得た。1.マウスストローマ細胞OP-9にNotchリガンドDelta1を発現させ、マウス造血幹細胞を共培養することにより、大量で均一なT前駆細胞が得られることを明らかにした。2.ヒト臍帯血中のCD133陽性細胞をマグネットビーズ法で純化し、サイトカインなどの存在下・無血清で培養したのち、重症免疫不全マウスへの移植実験を行うことにより、CD133陽性細胞が豊富に造血幹細胞を含み、これが無血清条件で増幅可能であることを明らかにした。3.ヒト臍帯血および末梢血のCD133陽性細胞を、Delta1を発現させたOP-9と共培養することにより、大量のT前駆細胞を産生しうることを見出した。今後、無血清条件、可溶型Delta1により同様にT前駆細胞を産生し、CD52モノクローナル抗体を用いたHLA不一致移植後に輸注する臨床試験に進むことを検討している。

Research Products

• [Journal Article] AML1 Is Functionally Regulated through p300-mediated Acetylation on Specific Lysine Residues. (2004)
  • Author(s): Yamaguchi Y, Chiba S, et al.
  • Journal Title: J Biol Chem 279 Pages: 15630-15638
• [Journal Article] Stem cell leukemia protein directs hematopoietic stem cell fate. (2004)
  • Author(s): Kunisato A, Chiba S, et al.
  • Journal Title: Blood 103 Pages: 3336-3341
• [Journal Article] AML-1 is required for megakaryocytic maturation and lymphocytic differentiation, but not for maintenance of hematopoietic stem cells in adult hematopoiesis. (2004)
  • Author(s): Ichikawa M, Chiba S, et al.
  • Journal Title: Nature Med 10 Pages: 299-304
• [Journal Article] Recovery of Valpha24(+) NKT cells after hematopoietic steal cell transplantation. (2004)
  • Author(s): Haraguchi K, Chiba S, et al.
  • Journal Title: Bone Marrow Transplant 34 Pages: 595-602
• [Journal Article] In vivo alemtuzumab enables haploidentical HLA-mismatched hematopoietic stem cell transplantation without ex vivo graft manipulation.
  • Author(s): Kanda Y, Chiba S, et al.
  • Journal Title: Transplantation (in press)
• [Journal Article] Graft-versus-tumor effect against advanced pancreatic cancer after allogeneic reduced-intensity stem cell transplantation.
  • Author(s): Kanda Y, Chiba S, et al.
  • Journal Title: Transplantation (in press)
• [Publications] Kunisato A: "Stem cell leukemia protein directs hematopoietic stem cell fate."Blood. (In Press). (2004)
• [Publications] Maekawa Y: "Delta1-Notch3 interactions bias the functional differentiation of activated CD4+ T cells."Immunity. 19. 549-559 (2004)
• [Publications] Takahashi T: "Valpha24+ natural killer T-cell responses against T-acute lymphoblastic leukaemia cells : implications for immunotherapy."Br J Haematol. 122. 231-239 (2003)
• [Publications] Takahashi T: "Dendritic cell vaccination for patients with chronic myelogenous leukemia."Leuk Res. 27. 795-802 (2003)
• [Publications] Saito T: "Notch2 Is Preferentially Expressed in Mature B Cells and Indispensable for Marginal Zone B Lineage Development."Immunity. 18. 675-685 (2003)
• [Publications] Kanda Y: "Allogeneic hematopoietic stem cell transplantation from family members other than HLA-identical siblings over the last decade."Blood. 102. 1541-1547 (2003)
• [Publications] Kunisato A, Chiba S, Nakagami-Yamaguchi E, Kumano K, Saito T, Masuda S, Yamaguchi T, Osawa M, Kageyama R, Nakauchi H, Nishikawa M, Hirai H.: "HES-1 preserves purified hematopoietic stem cells ex vivo and accumulates side population cells in vivo"Blood. 101. 1777-1783 (2003)
• [Publications] Kawazu M, Kanda Y, Goyama S, Takeshita M, Nannya Y, Niino M, Komeno Y, Nakamoto T, Kurokawa M, Tsujino S, Ogawa S, Aoki K, Chiba S, Motokura T, Ohishi N, Hirai H.: "Rapid diagnosis of invasive pulmonary aspergillosis by quantitative polymerase chain reaction using bronchial lavage fluid"Am J Hematol.. 72. 27-30 (2003)
• [Publications] Yamaguchi E, Chiba S, Kumano K, Kunisato A, Takahashi T, Hirai H.: "Expression of Notch ligands, Jagged1,2 and Deltal in antigen presenting cells in mice"Immunol Lett.. 81. 59-64 (2002)
• [Publications] Tanaka Y, Takahashi T, Nieda M, Masuda S, Kashiwase K, Ogawa S, Chiba S, Juji T, Hirai H.: "Generation of Fas-independent CD4+ cytotoxic T-cell clone specific for p190 minor bcr-abl fusion peptide"Leuk Res.. 26. 317-321 (2002)
• [Publications] Tanaka Y, Kanda Y, Kami M, Mori s, Hamaki T, Kusumi E, Miyakoshi S, Nannya Y, Chiba S, Arai Y, Mitani K, Hirai H, Mutou Y.: "Monitoring cytomegalovirus infection by antigenemia assay and two distinct plasma real-time PCR methods after hematopoietic stem cell transplantation"Bone Marrow Transplant. 30. 315-319 (2002)
• [Publications] Takahashi T, Chiba S, Nieda M, Azuma T, Ishihara S, Shibata Y, Juji T, Hirai H.: "Cutting edge : analysis of human v alpha 24+CD8+ NK T cells activated by alpha-galactosylceramide-pulsed monocyte-derived dendritic cells"J Immunol.. 168. 3140-3144 (2002)
• [Publications] Takahashi T, Chiba S, Matsumura T, Hirai H.: "A case of severe B cell deficiency after allogeneic stem cell transplantation"Bone Marrow Transplant. 30. 397-400 (2002)
• [Publications] Shimizu K, Chiba S, Saito T, Takahashi T, Kumano K, Hamada Y, Hirai H.: "Integrity of intracellular domain of Notch ligand is indispensable for cleavage required for release of the Notch2 intracellular domain"Embo J.. 21. 294-302 (2002)
• [Publications] Shimizu K, Chiba S, Saito T, Kumano K, Hamada Y, Hirai H.: "Functional diversity among Notch1, Notch2, Notch3 receptors"Biochem Biophys Res Commun.. 291. 775-779 (2002)
• [Publications] Saito A, Ogawa S, Mori S, Sakamaki H, Kami M, Miyakoshi J, Muto Y, Hatama T, Kawano M, Kinoshita M, Chiba S, Hirai H.: "Establishment of a quantitative method for detecting mixed chimerism using fluorescence-based PCR amplification of short tandem repeat markers after allogeneic hematopoietic stem cell transplantation in a Japanese population"Rinsho Ketsueki. 43. 327-332 (2002)
• [Publications] Imai O, Kurokawa M, Izutsu K, Hangaishi A, Maki K, Ogawa S, Chiba S, Mitani K, Hirai H.: "Mutational analyses of the AML1 gene in patients with myelodysplastic syndrome"Leuk Lymphoma. 43. 617-621 (2002)
• [Publications] Goyama S, Kanda Y, Nannya Y, Kawazu M, Takeshita M, Niino M, Komeno Y, Nakamoto T, Kurokawa M, Tsujino S, Ogawa S, Aoki K, Chiba S, Motokura T, Shiratori Y, Hirai H.: "Reverse seroconversion of hepatitis B virus after hematopoietic stem cell transplantation"Leuk Lymphoma. 43. 2159-2163 (2002)
• [Publications] Chiba S, Saito A, Ogawa S, Takeuchi K, Kumano K, Seo S, Suzuki T, Tanaka Y, Saito T, Izutsu K, Yuji K, Masuda S, Futami s, Nishida M, Suzuki G, Gale RP, Fukayama M, Maekawa K, Hirai H.: "Transplantation for accidental acute high-dose total body neutron-and gamma-radiation exposure"Bone Marrow Transplant. 29. 935-939 (2002)
• [Publications] Takahashi T, Chiba S, Matsumura T, Hirai H.: "A case of severe B cell deficiency after allogeneic stem cell transplantation"Bone Marrow Transplant. 30. 397-400 (2002)
• [Publications] Shimizu K, Chiba S, Saito T, Takahashi T, Kumano K, Hamada Y, Hirai H.: "Integrity of intracellular domain of Notch ligand is indispensable for cleavage required for release of the Notch2 intracellular domain"Embo J.. 21. 294-302 (2002)
• [Publications] Shimizu K, Chiba S, Saito T, Kumano K, Hamada Y, Hirai H.: "Functional diversity among Notch1, Notch2, and Notch3 receptors"Biochem Biophys Res Commun.. 291. 775-779 (2002)
• [Publications] Takahashi, T.: "Analysis of human Va24+CD8+ natural killer T cells activated by a-galactosylceramide-pulsed monocyte-derived dendritic cells"J. Immunol. (in press).
• [Publications] Chiba, S.: "Transplantation for Accidental Acute High-Dose Total Body Neutron-and γ -Radiation Exposure"Bone Marrow Transplant. (in press).
• [Publications] Tanaka, Y.: "Generation of Fas-Independent CD4+ cytotoxic T-cell clone specific for p190 minor bcr-ab1 fusion peptide"Leukemia Res. 26. 317-321 (2002)
• [Publications] Yamaguchi, E.: "Expression of Nocth ligand, Jagged1, Jagged2 and Delta1 in antigen-presenting cells in mice"Immunol. Letter. 81. 59-64 (2002)
• [Publications] Shimizu, K.: "Integrity of intracellular domain of Notch ligand is indispensable for cleavage required for release of Notch2 intracellular domain"EMBO J.. 21. 294-302 (2002)
• [Publications] Kumano, K.: "Notch1 inhibits differentiation of hematopoietic cells by sustaining GATA-2 expression"Blood. 98. 3283-3289 (2001)