1998 Fiscal Year Annual Research Report

マウスAGM領域からの造血幹細胞自己複製因子のクローニング

Research Project

Project/Area Number	09357007
Research Institution	The University of Tokyo
Principal Investigator	中畑龍俊東京大学, 医科学研究所, 教授 (20110744)
Co-Investigator(Kenkyū-buntansha)	渡辺すみ子東京大学, 医科学研究所, 助手 (60240735) 前川平東京大学, 医科学研究所, 講師 (80229286) 辻浩一郎東京大学, 医科学研究所, 助教授 (50179991)
Keywords	造血幹細胞 / 自己複製因子 / マウス / AGM領域 / クローニング / サイトカイン / 臨床応用 / NOD / SCIDマウス
Research Abstract	マウス造血幹細胞は胎生初期に大動脈(aorta)、性腺(gonad)、中腎(mesonephros)に囲まれた領域(AGM領域)で発生、増幅し、その後肝臓へ移動していくことが報告された。AGM領域は造血組織ではなく、ここでは造血幹細胞が発生、爆発的に増殖するものの分化を伴わなわず、造血幹細胞の自己複製因子が強く発現している可能性が示唆されることから、AGM領域から造血幹細胞の増殖支持能を有するストローマ細胞株の樹立を試みた。胎生9.5,10,10.5,11,11.5,12日目の胎児からAGM領域を取り出し種々の条件で培養した。血液細胞、筋肉、神経など様々な細胞の増殖が見られたが、その中からストローマ細胞の増殖の得られた培養を選択し、さらに限界希釈法を用いて造血幹細胞の増幅支持能を持ったストローマ細胞株の樹立を試み、ついに成功した。AGM領域からストローマ細胞株の樹立に成功したのはわれわれが世界で始めてである。われわれの樹立したAGM-S3上にヒトCD34+CD38-細胞を重層して培養すると、サイトカインの非存在下に8週間以上未分化造血前駆細胞を産生し続けることが明らかとなった。また、われわれがヒト造血幹細胞のin vivo測定系として開発したNOD/SCIDマウスを用いた検討では、NOD/SCIDマウス骨髄を再構築する能力を持ったヒト造血幹細胞が、AGM-S3上で4週以上にわたって増殖し続けていることが明らかとなった。また、造血支持能を持たない細胞株AGM-A7も同時にクローニングした。予備的な検討では、AGM-S3はIL-6,SCF,EPO,OSM,MIP-1α,EPO mRNAを発現しているものの、これらのマウスサイトカインはヒト造血幹細胞の増殖に作用しないことから、AGM-S3は種を越えて造血幹細胞の自己複製に働く未知の分子を発現していると考えられる。現在、造血幹細胞増幅支持能を持ったAGM-S3および支持能を持たないAGM-A7のそれぞれからmRNAを抽出し、一方向のcDNAライブラリーを作製し、サブトラクション法、発現クローニング法などを用いて造血幹細胞自己複製因子の遺伝子クローニングを開始している。

Research Products

(27 results)

All Other

All Publications (27 results)

[Publications] Manabe A., MORI T., Ebihara Y., Nakahata T., et al: "Characterization of leukemic cells in CD2/CD19 double positive acute lymphoblastic leukemia." Int.J.Hematol.67. 45-52 (1998)
[Publications] Mukouyama Y., Hara T., Xu M., Nakahata T., et al: "Induction of hematopoiesis by oncostatin M in the AGM region." Immunity. 8. 105-114 (1998)
[Publications] Toru H., Eguchi M., Yata J., Nakahata T., et al: "IL-4 promotes the development of tryptase and chymase double positive human mast cells accompanied with cell maturation." Blood. 91. 187-195 (1998)
[Publications] Nakahata T.: "Characteristics of hemopoietic stem/progenitor cells in bone marrow, peripheral blood and cord blood." Int.J.Pediatr.Hematol.Oncology. 5. 60-61 (1998)
[Publications] Takahashi T., Yamada K., Nakahata T., Hirai H., et al: "A novel molecular approach to ex vivo hematopoietic expansion with recombinant EGFR-expressing adenovirus vector." Blood. 91. 4509-4515 (1998)
[Publications] Nishihara M., Wada Y., Nakahata T., Maekawa T., et al: "A combination of stem cell factor and granulocyte colony-stimulating factor enhances the growth of human progenitor B cells supported by murine stromal cell line MS-5." Eur.J.Immunol.28. 855-864 (1998)
[Publications] Tsuji K., Muraoka K., Nakahata T., et al: "Interferon-g and human megakaryopoiesis." Leukemia and Lymphoma. 31. 107-113 (1998)
[Publications] Tajika K., Ikebuchi K., Nakahata T., Asano S., et al: "IL-6 and SCF exart different effects on megakaryocyte maturation." Brit.J.Haematol.100. 105-111 (1998)
[Publications] Ishiguro A., Ishikita T., Naritake S., Nakahata T., et al: "Elevation of serum thrombopoietin precedes thrombocytopoiesis in Kawasaki disease." Thromb.Haemostasis. 79. 1096-1100 (1998)
[Publications] Yoshida K., Taga T., Nakahata T., Kishimoto T., et al: "Myocardial, hematological, and placental disorders caused by targeted disruption of gp130, acommon signal transducer for IL-6 family of cytokines." Contemporary Immunology. 259-289 (1998)
[Publications] Suzuki H., Takei M., Nakahata T., Fukamachi H., et al: "Inhibitory effect of adenosine on degranulation of human cultured mast cells upon cross-linking of FceRI." Biochem.Biophy.Res.Comm.242. 697-702 (1998)
[Publications] Sato T., Watanabe S., Tsuji K., Nakahata T., et al: "Induction of the erythropoietin receptor gene and acquisition of responsiveness to erythropoietin by stem cell factor in HML/SE, a human leukemic cell line." J.Biol.Chem.273. 16921-16926 (1998)
[Publications] Maruyama K., Tsuji K., Kodera Y., Nakahata T., et al: "Characterization of peripheral blood progenitor cells mobilized by Nartograstim (N-terminal replaced granulocyte colony-stimulating factor) in normal volunteers." Bone Marrow Transplant.22. 313-320 (1998)
[Publications] Xu M., Tsuji K., Miyajima A., Nakahata T., et al: "Stimulation of mouse and human primitive hematopoiesis by murine embryonic aorta-gonad-mesonephros-derived stromal cells." Blood. 192. 2032-2040 (1998)
[Publications] Shichijo M., Nakahata T., Saito H., Nagai H., et al: "The effect of anti-asthma drags on mediator release from cultured human mast cells." Clin.Exp.Allergy. 28. 1229-1236 (1998)
[Publications] Sako M., Nakahaat T., Kishitomo T., Sugiyama H., et al: "Abnormal expression of Wilms'tumor gene WT1 in juvenile chronic myeloid leukemia and infantile monosomy 7 syndrome." Leukemia Res.22. 965-967 (1998)
[Publications] Toru H., Pawankar R., Yata J., Nakahata T., et al: "Human mast cells produce interleukin-13 by high affinity IgE receptor cross-linking : enhanced IL-13 production by IL-4 primed human mast cells." J.Allergy and Clin.Immunol.(in press.). (1998)
[Publications] Yang FC, Watanabe S., Ebihary Y., Nakahata T., et al: "Human G-CSF stimulates the development of primitive multipotential progenitors of human G-CSF receptor-transgenic mice, but does not affect their commitment in vitro and in vivo." Blood. 92. 1632-1640 (1998)
[Publications] Nakahata T., Sui X., Kishimoto T., et al: "Ex vivo expansion of human primitive hemopoietic progenitors." Stem Cells.(in press.). (1998)
[Publications] Sui X., Tsuji K., Kishimoto T., Nakahata T., et al: "Soluble IL-6 receptor with IL-6 stimulates megakaryopoiesis from human CD34+cells through gp130 signaling." Blood. (in press.). (1998)
[Publications] Yang FC., Tsuji K., Ikeda Y., Nakahata T., et al: "Effects of human granulocyte colony-stimulating factor (hG-CSF) on megakaryopoiesis and platelet function in hG-CSF receptor-transgenic mice." Blood. (in press.). (1998)
[Publications] Taniguchi T., Endo H., Nakahata T., Motokura T., et al: "Expression of p21Cip1/Waf1/Sdi1 and p27 Kip1 cyclin-dependent kinase inhibitors during human hematopoiesis." Blood. (in press.). (1998)
[Publications] Duraisamy K., Saito H., Tsuji K., Nakahata T., et al: "Characterization of mast-cell-committed progenitors present in human cord blood." Blood. (in press.). (1998)
[Publications] Nakahata T., Toru H., et at: "Development of human mast cells." Adv.Immunol.(in press.). (1998)
[Publications] Hasegawa S., Nakahata T., Okumura K., Ra C., et al: "Functional expression of the high affinity receptor for IgE (Fc epsilon RI)in human platelets and its intracellular expression in human megakaryocytes." Blood. (in press.). (1998)
[Publications] Hibino H., Tani K., Nakahata T., Asano S., et al: "Common Mamoset as a target preclinical primate for cytokine and gene therapy studies." Blood. (in press.). (1998)
[Publications] Kobayashi M., Ueda K., Nishihira H., Nakahata T., et al: "Serum granulocyte colony-stimulating factor levels in patients with chronic neutropenia of childhood : Modulation of G-CSF levels by myeloid precursor cell mass." Brit.J.Haematol.(in press.). (1998)

1998 Fiscal Year Annual Research Report

マウスAGM領域からの造血幹細胞自己複製因子のクローニング

Principal Investigator

中畑 龍俊 東京大学, 医科学研究所, 教授 (20110744)

Research Products

[Publications] Manabe A., MORI T., Ebihara Y., Nakahata T., et al: "Characterization of leukemic cells in CD2/CD19 double positive acute lymphoblastic leukemia." Int.J.Hematol.67. 45-52 (1998)

[Publications] Mukouyama Y., Hara T., Xu M., Nakahata T., et al: "Induction of hematopoiesis by oncostatin M in the AGM region." Immunity. 8. 105-114 (1998)

[Publications] Toru H., Eguchi M., Yata J., Nakahata T., et al: "IL-4 promotes the development of tryptase and chymase double positive human mast cells accompanied with cell maturation." Blood. 91. 187-195 (1998)

[Publications] Nakahata T.: "Characteristics of hemopoietic stem/progenitor cells in bone marrow, peripheral blood and cord blood." Int.J.Pediatr.Hematol.Oncology. 5. 60-61 (1998)

[Publications] Takahashi T., Yamada K., Nakahata T., Hirai H., et al: "A novel molecular approach to ex vivo hematopoietic expansion with recombinant EGFR-expressing adenovirus vector." Blood. 91. 4509-4515 (1998)

[Publications] Nishihara M., Wada Y., Nakahata T., Maekawa T., et al: "A combination of stem cell factor and granulocyte colony-stimulating factor enhances the growth of human progenitor B cells supported by murine stromal cell line MS-5." Eur.J.Immunol.28. 855-864 (1998)

[Publications] Tsuji K., Muraoka K., Nakahata T., et al: "Interferon-g and human megakaryopoiesis." Leukemia and Lymphoma. 31. 107-113 (1998)

[Publications] Tajika K., Ikebuchi K., Nakahata T., Asano S., et al: "IL-6 and SCF exart different effects on megakaryocyte maturation." Brit.J.Haematol.100. 105-111 (1998)

[Publications] Ishiguro A., Ishikita T., Naritake S., Nakahata T., et al: "Elevation of serum thrombopoietin precedes thrombocytopoiesis in Kawasaki disease." Thromb.Haemostasis. 79. 1096-1100 (1998)

[Publications] Yoshida K., Taga T., Nakahata T., Kishimoto T., et al: "Myocardial, hematological, and placental disorders caused by targeted disruption of gp130, acommon signal transducer for IL-6 family of cytokines." Contemporary Immunology. 259-289 (1998)

[Publications] Suzuki H., Takei M., Nakahata T., Fukamachi H., et al: "Inhibitory effect of adenosine on degranulation of human cultured mast cells upon cross-linking of FceRI." Biochem.Biophy.Res.Comm.242. 697-702 (1998)

[Publications] Sato T., Watanabe S., Tsuji K., Nakahata T., et al: "Induction of the erythropoietin receptor gene and acquisition of responsiveness to erythropoietin by stem cell factor in HML/SE, a human leukemic cell line." J.Biol.Chem.273. 16921-16926 (1998)

[Publications] Maruyama K., Tsuji K., Kodera Y., Nakahata T., et al: "Characterization of peripheral blood progenitor cells mobilized by Nartograstim (N-terminal replaced granulocyte colony-stimulating factor) in normal volunteers." Bone Marrow Transplant.22. 313-320 (1998)

[Publications] Xu M., Tsuji K., Miyajima A., Nakahata T., et al: "Stimulation of mouse and human primitive hematopoiesis by murine embryonic aorta-gonad-mesonephros-derived stromal cells." Blood. 192. 2032-2040 (1998)

[Publications] Shichijo M., Nakahata T., Saito H., Nagai H., et al: "The effect of anti-asthma drags on mediator release from cultured human mast cells." Clin.Exp.Allergy. 28. 1229-1236 (1998)

[Publications] Sako M., Nakahaat T., Kishitomo T., Sugiyama H., et al: "Abnormal expression of Wilms'tumor gene WT1 in juvenile chronic myeloid leukemia and infantile monosomy 7 syndrome." Leukemia Res.22. 965-967 (1998)

[Publications] Toru H., Pawankar R., Yata J., Nakahata T., et al: "Human mast cells produce interleukin-13 by high affinity IgE receptor cross-linking : enhanced IL-13 production by IL-4 primed human mast cells." J.Allergy and Clin.Immunol.(in press.). (1998)

[Publications] Yang FC, Watanabe S., Ebihary Y., Nakahata T., et al: "Human G-CSF stimulates the development of primitive multipotential progenitors of human G-CSF receptor-transgenic mice, but does not affect their commitment in vitro and in vivo." Blood. 92. 1632-1640 (1998)

[Publications] Nakahata T., Sui X., Kishimoto T., et al: "Ex vivo expansion of human primitive hemopoietic progenitors." Stem Cells.(in press.). (1998)

[Publications] Sui X., Tsuji K., Kishimoto T., Nakahata T., et al: "Soluble IL-6 receptor with IL-6 stimulates megakaryopoiesis from human CD34+cells through gp130 signaling." Blood. (in press.). (1998)

[Publications] Yang FC., Tsuji K., Ikeda Y., Nakahata T., et al: "Effects of human granulocyte colony-stimulating factor (hG-CSF) on megakaryopoiesis and platelet function in hG-CSF receptor-transgenic mice." Blood. (in press.). (1998)

[Publications] Taniguchi T., Endo H., Nakahata T., Motokura T., et al: "Expression of p21Cip1/Waf1/Sdi1 and p27 Kip1 cyclin-dependent kinase inhibitors during human hematopoiesis." Blood. (in press.). (1998)

[Publications] Duraisamy K., Saito H., Tsuji K., Nakahata T., et al: "Characterization of mast-cell-committed progenitors present in human cord blood." Blood. (in press.). (1998)

[Publications] Nakahata T., Toru H., et at: "Development of human mast cells." Adv.Immunol.(in press.). (1998)

[Publications] Hasegawa S., Nakahata T., Okumura K., Ra C., et al: "Functional expression of the high affinity receptor for IgE (Fc epsilon RI)in human platelets and its intracellular expression in human megakaryocytes." Blood. (in press.). (1998)

[Publications] Hibino H., Tani K., Nakahata T., Asano S., et al: "Common Mamoset as a target preclinical primate for cytokine and gene therapy studies." Blood. (in press.). (1998)

[Publications] Kobayashi M., Ueda K., Nishihira H., Nakahata T., et al: "Serum granulocyte colony-stimulating factor levels in patients with chronic neutropenia of childhood : Modulation of G-CSF levels by myeloid precursor cell mass." Brit.J.Haematol.(in press.). (1998)

中畑龍俊東京大学, 医科学研究所, 教授 (20110744)