1999 Fiscal Year Annual Research Report

サイトカインレセプタートランスジェニックマウスを用いた造血幹細胞の分化機構の解析

Research Project

Project/Area Number	10307020
Research Institution	KYOTO UNIVERSITY
Principal Investigator	中畑龍俊京都大学, 大学院・医科学研究科, 教授 (20110744)
Co-Investigator(Kenkyū-buntansha)	依藤亨京都大学, 大学院・医学研究科, 助手 (60220779) 辻浩一郎東京大学, 医学研究所, 助教授 (50179991)
Keywords	造血幹細胞 / サイトカイン / レセプター / トランスジェニック / GM-CSFR / G-CSFR / マウス / 分化
Research Abstract	造血幹細胞の増殖・分化におけるサイトカインの役割を明らかにするため、GM-CSF,G-CSFの受容体を発現するトランスジェニツク(Tg)マウスを作成した。 hGM-CSFR-Tgマウス骨髄細胞をGM-CSF存在下で培養すると単球、好中球系のみならず未分化、赤芽球系、巨核球系、肥満細胞系の前駆細胞の増殖と最終的な分化、成熟が得られた。また、TgマウスヘのGM-CSFの投与により赤血球、好中球、単球、NK細胞の著明な増加が観察されている。GM-CSF単独で赤血球産生が起こることから、これがEPORを介するか否か検討するため、EPOR-/-マウスとGM-CSFR Tgマウスを掛け合わせて検討した。EPOR-/-xGM-CSFR Tgは胎生14日前後で死亡したが、このマウスから胎児肝を取りだし、GM-CSF存在下で培養すると多数のCFU-Eコロニーが形成された。以上の結果はEPORシグナルとは全く関係なく受容体さえ発現していればGM-CSFシグナルによっても赤血球産生が起こることを示している。hG-CSF-Tgマウスでも同様にG-CSF刺激により様々な前駆細胞の増殖・分化・成熟が見られたが、CSF単独での赤血球までの最終分化は見られなかった。以上のG-CSFの作用はFACSでソーティングした細胞の単細胞培養でも確認された。さらに、ヒトG-CSF存在下で形成された芽球コロニーを取り出して、ヒトG-CSF存在下で培養すると、再びさまざまな血球系のコロニーが形成された.以上よりGM-CSFやG-CSFは多能性造血幹細胞から好中球、マクロファージヘの分化を決定する因子として作用するのではなく、受容体を発現している細胞に単なる増殖因子として働き、全ての血球の分化がもたらされることを示唆している。

Research Products

(23 results)

All Other

All Publications (23 results)

[Publications] Sui X.,Tsuji K.,Ebihara Y.,Tkishimoto T.,Nakahata T.: "Soluble IL-6 receptor with IL-6 stimulates megakaryopoiesis from human CD34_+ cells through gp130 signaling"Blood. 93. 2525-2532 (1999)
[Publications] Hasegawa S.,Pawankar R.,Suzaki K.,Nakahata T.,Furukawa S.,Okumura K.,Ra C.: "Functional expression of the high affinity receptor for IgE (Fc epsilon RI) in human Platelets and its intracellular expression in human megakaryocytes"Blood. 93. 2543-2551 (1999)
[Publications] Honda H.,Suzuki T.,Inaba T.,Tsuji K.Nakahata T.,Yazaki Y.,Look T.,Hirai H.: "Expression of E2a-HLF chineric protein induced T-cell apotosis, B-cell maturation arrest, and development of acute lymphoblastic leukemia"Blood. 93. 2780-2790 (1999)
[Publications] Kampuraj D.,Saito H.,Kaneko A.,Fukagawa K.,Tsuji K.,Nakahata T.: "Characterization of mast-cell-committed progenitors present in human cord blood"Blood. 93. 3338-3346 (1999)
[Publications] Kobayashi M.,Ueda K.,Kojima S.,Shimbo T.,Nishihira H.,Nakahata T.: "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.. 105. 486-490 (1999)
[Publications] Taniguchi T.,Endo H.,Asano S.,Fujita T.,Nakahata T.,Motokura T.: "Expression of p21Cip1/Waf1/Sdil and p27 Kip1 cyclin-dependent kinase inhibitors during human hematopoiesis"Blood. 83. 4167-4178 (1999)
[Publications] Kinoshita T.,Sekiguchi T.,Xu M-J.,Ito Y.,Tsuji K.,Nakahata T.,Miyajima A.: "Hepatic differentiation induced by oncostatin M attenuates fetal liver hematopoiesis"Proc, Natl. Acad. Sci. USA. 96. 7265-7270 (1999)
[Publications] Yang FC.,Tsuji K.,Oda A.,Xu MJ.,Ebihara Y.,Watanabe S.,Ikeda Y.,Nakahata T.: "Effects of human granuloyle colony-stimulating factor (hG-CSF) on megakaryopoiesis and platelet function in hG-CSF receptor-transgenic mice"Blood. 94. 950-958 (1999)
[Publications] Matsumoto T.,Seki Y.,Kubo M.,Nakahata T.,Yamada S.,Yoshimura A.: "Suppression of STAT5 functions in liver,mammary grands, and T cells in cytokine-inducible SH2 protein-1 (CIS1) transgenic mice"Mol. Cell. Biol.. 19. 6396-6407 (1999)
[Publications] Ui M.,Takeda M.,Arai T.,Yamada K.,Nakahata T.,Tokino T.,Nakamura Y.,Iba H.: "Retrovirus vectors designed for efficient transduction of cytotoxic or cytostatic genes"Gene Therapy. 6. 1670-1678 (1999)
[Publications] Ishii T.,Nakahata T.,Maekawa T.: "Expression of SDF-1/PBSF receptor CXCR4 on CD34+ human bone marrow cells a phenotypic alteration for committed lymphoid progenitors"J. Immunol.. (in press).
[Publications] Kajiwara M.,Nonoyama S.,Eguchi M.,Imai K.,Ochs HD.,Nakahata T.,Yata J.: "In vitro megakaryopocyt differentiation from CD34 positive cells is defective in patients with Wiscott-Aldrich syndrome and X-linked thrombocytopenia"Brit. J. Haematol.. (in pres).
[Publications] Takagi M.,Nakamura T.,Sawada T.,Nakahata T.,Yokota T.,Heike T.: "Chimeric GM-CSF-leukemia inhibitory factor (LIF) receptor can transduce expansion signals in primitive hematopoietic progenitors lacking IL-6Ra, IL-11Ra and gp130"Mol.Cell.Biol.. (in press).
[Publications] Ishiguro A.,Nakahata T.,Kato T.,Suzuki Y.,Shimbo T.: "Age-related changes in serum thrombopoietin in children:high levels in infants by one month of age"Pediatr, Res.. (in press).
[Publications] Futaki M.,Yamashita T.,Yabe M.,Kato S.,Asano S.,Nakahata T.: "The IVS4+4A-T mutation of the Fanconi Anemia gene FANCC is associated with severe phenotype in Japanese patients"Blood. (in press).
[Publications] Matsuoka S.,Tsuji K.,Xu M.,Yang FC.,Kaneko K.,Kikuchi A.,Nakahata T.: "CD 34 expression on long-term repopulating hematopoietic stem cells changes during developmental stages"Blood. (in press).
[Publications] Ueda T.,Yoshino H.,Ebihara Y.,Ito M.,Tsuji K.,Nakahata T: "Ex vivo expansion of human NOD/SCID-repopulating cells by stem cell factor, Flk2/Flt3 ligand, thrombopoietin interleukin-6 and soluble interleukin-6 receptor"J. Clin. Invest.. (in press).
[Publications] Hayashi Y.,Honma Y.,Niitsu N.,Taki T.,Tsuji K.,Nakahata T.: "SN-1, a novel leukemic cell line with t(11;16) (q23;p13): Myeloid characteristics and resistance to retinoids and viamin D3"Cancer Res.. (in press).
[Publications] Sugiyama H.,Nonaka T.,Kishimoto T.,Komoriya K.,Tsuji K.,Nakahata T.: "Peroxisome prolifertor-activated receptors negatively regurate mast cell activation"FEBS Letters. (in press).
[Publications] Sugiyama H.,Nonaka T.,Kishimoto T.,Komoriya K.,Tsuji K.,Nakahata T.: "Peroxisome proliferator-activated receptors are expressed in human cultured mast cells:a possible role for these receptors in negative regulation of mast cell activation"Blood. (in press).
[Publications] 中畑龍俊: "In vitro での末梢血幹細胞の増幅.症例に学ぶ G-CSF の臨床"医薬ジャーナル. 14 (1999)
[Publications] 中畑龍俊、二木真琴: "遺伝子治療開発研究ハンドブック"日本遺伝子治療学会編、エヌ・テイ・エス. 6 (1999)
[Publications] 辻浩一郎、中畑龍俊: "内科学教科書 (印刷中)"文光堂(東京).

1999 Fiscal Year Annual Research Report

サイトカインレセプタートランスジェニックマウスを用いた造血幹細胞の分化機構の解析

Principal Investigator

中畑 龍俊 京都大学, 大学院・医科学研究科, 教授 (20110744)

Research Products

[Publications] Sui X.,Tsuji K.,Ebihara Y.,Tkishimoto T.,Nakahata T.: "Soluble IL-6 receptor with IL-6 stimulates megakaryopoiesis from human CD34_+ cells through gp130 signaling"Blood. 93. 2525-2532 (1999)

[Publications] Hasegawa S.,Pawankar R.,Suzaki K.,Nakahata T.,Furukawa S.,Okumura K.,Ra C.: "Functional expression of the high affinity receptor for IgE (Fc epsilon RI) in human Platelets and its intracellular expression in human megakaryocytes"Blood. 93. 2543-2551 (1999)

[Publications] Honda H.,Suzuki T.,Inaba T.,Tsuji K.Nakahata T.,Yazaki Y.,Look T.,Hirai H.: "Expression of E2a-HLF chineric protein induced T-cell apotosis, B-cell maturation arrest, and development of acute lymphoblastic leukemia"Blood. 93. 2780-2790 (1999)

[Publications] Kampuraj D.,Saito H.,Kaneko A.,Fukagawa K.,Tsuji K.,Nakahata T.: "Characterization of mast-cell-committed progenitors present in human cord blood"Blood. 93. 3338-3346 (1999)

[Publications] Kobayashi M.,Ueda K.,Kojima S.,Shimbo T.,Nishihira H.,Nakahata T.: "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.. 105. 486-490 (1999)

[Publications] Taniguchi T.,Endo H.,Asano S.,Fujita T.,Nakahata T.,Motokura T.: "Expression of p21Cip1/Waf1/Sdil and p27 Kip1 cyclin-dependent kinase inhibitors during human hematopoiesis"Blood. 83. 4167-4178 (1999)

[Publications] Kinoshita T.,Sekiguchi T.,Xu M-J.,Ito Y.,Tsuji K.,Nakahata T.,Miyajima A.: "Hepatic differentiation induced by oncostatin M attenuates fetal liver hematopoiesis"Proc, Natl. Acad. Sci. USA. 96. 7265-7270 (1999)

[Publications] Yang FC.,Tsuji K.,Oda A.,Xu MJ.,Ebihara Y.,Watanabe S.,Ikeda Y.,Nakahata T.: "Effects of human granuloyle colony-stimulating factor (hG-CSF) on megakaryopoiesis and platelet function in hG-CSF receptor-transgenic mice"Blood. 94. 950-958 (1999)

[Publications] Matsumoto T.,Seki Y.,Kubo M.,Nakahata T.,Yamada S.,Yoshimura A.: "Suppression of STAT5 functions in liver,mammary grands, and T cells in cytokine-inducible SH2 protein-1 (CIS1) transgenic mice"Mol. Cell. Biol.. 19. 6396-6407 (1999)

[Publications] Ui M.,Takeda M.,Arai T.,Yamada K.,Nakahata T.,Tokino T.,Nakamura Y.,Iba H.: "Retrovirus vectors designed for efficient transduction of cytotoxic or cytostatic genes"Gene Therapy. 6. 1670-1678 (1999)

[Publications] Ishii T.,Nakahata T.,Maekawa T.: "Expression of SDF-1/PBSF receptor CXCR4 on CD34+ human bone marrow cells a phenotypic alteration for committed lymphoid progenitors"J. Immunol.. (in press).

[Publications] Kajiwara M.,Nonoyama S.,Eguchi M.,Imai K.,Ochs HD.,Nakahata T.,Yata J.: "In vitro megakaryopocyt differentiation from CD34 positive cells is defective in patients with Wiscott-Aldrich syndrome and X-linked thrombocytopenia"Brit. J. Haematol.. (in pres).

[Publications] Takagi M.,Nakamura T.,Sawada T.,Nakahata T.,Yokota T.,Heike T.: "Chimeric GM-CSF-leukemia inhibitory factor (LIF) receptor can transduce expansion signals in primitive hematopoietic progenitors lacking IL-6Ra, IL-11Ra and gp130"Mol.Cell.Biol.. (in press).

[Publications] Ishiguro A.,Nakahata T.,Kato T.,Suzuki Y.,Shimbo T.: "Age-related changes in serum thrombopoietin in children:high levels in infants by one month of age"Pediatr, Res.. (in press).

[Publications] Futaki M.,Yamashita T.,Yabe M.,Kato S.,Asano S.,Nakahata T.: "The IVS4+4A-T mutation of the Fanconi Anemia gene FANCC is associated with severe phenotype in Japanese patients"Blood. (in press).

[Publications] Matsuoka S.,Tsuji K.,Xu M.,Yang FC.,Kaneko K.,Kikuchi A.,Nakahata T.: "CD 34 expression on long-term repopulating hematopoietic stem cells changes during developmental stages"Blood. (in press).

[Publications] Ueda T.,Yoshino H.,Ebihara Y.,Ito M.,Tsuji K.,Nakahata T: "Ex vivo expansion of human NOD/SCID-repopulating cells by stem cell factor, Flk2/Flt3 ligand, thrombopoietin interleukin-6 and soluble interleukin-6 receptor"J. Clin. Invest.. (in press).

[Publications] Hayashi Y.,Honma Y.,Niitsu N.,Taki T.,Tsuji K.,Nakahata T.: "SN-1, a novel leukemic cell line with t(11;16) (q23;p13): Myeloid characteristics and resistance to retinoids and viamin D3"Cancer Res.. (in press).

[Publications] Sugiyama H.,Nonaka T.,Kishimoto T.,Komoriya K.,Tsuji K.,Nakahata T.: "Peroxisome prolifertor-activated receptors negatively regurate mast cell activation"FEBS Letters. (in press).

[Publications] Sugiyama H.,Nonaka T.,Kishimoto T.,Komoriya K.,Tsuji K.,Nakahata T.: "Peroxisome proliferator-activated receptors are expressed in human cultured mast cells:a possible role for these receptors in negative regulation of mast cell activation"Blood. (in press).

[Publications] 中畑龍俊: "In vitro での末梢血幹細胞の増幅.症例に学ぶ G-CSF の臨床"医薬ジャーナル. 14 (1999)

[Publications] 中畑龍俊、二木真琴: "遺伝子治療開発研究ハンドブック"日本遺伝子治療学会編、エヌ・テイ・エス. 6 (1999)

[Publications] 辻浩一郎、中畑龍俊: "内科学教科書 (印刷中)"文光堂(東京).

中畑龍俊京都大学, 大学院・医科学研究科, 教授 (20110744)