1997 Fiscal Year Annual Research Report

ヒトG-CSFレセプタートランスジェニックマウスを用いた白血病発症機構の研究

Research Project

Project/Area Number	09254210
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) 辻浩一郎東京大学, 医科学研究所, 助教授 (50179991) 岩倉洋一郎東京大学, 医科学研究所, 教授 (10089120)
Keywords	G-CSF受容体(G-CSFR) / Kostmann症候群 / 変異G-CSFR / 白血病 / MDS / トランスジェニックマウス / 造血幹細胞 / 血球分化
Research Abstract	Kostmann症候群の一部の患者ではG-CSFR受容体(G-CSFR)遺伝子の変異が見られ、G-CSFの長期使用により骨髄異形成症候群(MDS)や急性白血病が高頻度に合併することが知られている。G-CSFRの変異と急性白血病発症との関係を明らかにするため、ヒト、マウスwild typeおよびKostmann症候群で見られる変異G-CSFR遺伝子を発現する4種類のトランスジェニックマウス(Tgマウス)の作成を試みている。既にwild typeのヒト及びマウスG-CSFR cDNAに発現調節ユニットを連結したプラスミドを構築し、マウス線維芽細胞へのトランスフェクションによりG-CSFR遺伝子の発現をFACSにて確認した。マイクロインジェクション用にまずヒトG-CSFR DNAを調整し、C3H/HeN系マウス受精卵に注入後、偽妊娠マウスの輸卵管に計150個移植し、12匹のマウスにhG-CSFR遺伝子が確認された。hG-CSFRの発現が見られたマウスを交配し得られたF1マウス骨髄細胞を用いて、hG-CSFR蛋白の発現をFACSを用いて確認したところ、2系統のwild type hG-CSFRTgマウスがえられた。hG-CSFR-Tgマウスおよびそのリッターメイトより骨髄、脾細胞を採取し、メチールセルロース培養法を用いて造血幹細胞・造血前駆細胞の増殖・分化に対するG-CSFの作用を検討した。Tgマウス骨髄細胞をヒトG-CSF存在下で培養すると、顆粒球・マクロファージばかりではなく巨核球、肥満細胞、芽球コロニーの形成が見られた。G-CSF存在下で形成された芽球コロニーを取り出し再培養すると、再び種々の血球系コロニーが形成された。以上の結果よりG-CSFが造血幹細胞から顆粒球への分化の方向づけを決定する因子として作用するのではなく、G-CSF受容体を発現している細胞に単なる増殖因子として働くことを示唆された。TgマウスにヒトG-CSFを投与すると、好中球のみならず血小板など他の血球も増加した。G-CSFを連続投与後3ヵ月経過した現在、wild type hG-CSFRTgマウスからはMDSや急性白血病は見られていない。

Research Products

(28 results)

All Other

All Publications (28 results)

[Publications] Muraoka K.: "Defective response to thrombopoietin and impared expression of c-mpl mRAD in bone marrow cells in congenital amegakaryocytic thrombocytopenia." Brit.J.Haematol.96. 287-292 (1997)
[Publications] Eguchi M.: "Morphological and cytochemical changes in human mast cells during culture." Med.Electron Microsc.30. 25-30 (1997)
[Publications] Toru H.: "Interleukin-4 induces homotypic aggregation of human mast cells by promoting LFA-1/ICAM-1 adhesion molecules." Blood. 89. 3296-3302 (1997)
[Publications] Yanagida M.: "Effect of a chymotrypsin-like inhibitor,TPCK,on histamine release from cultured human mast cells." J.Pharm.Pharmth.49. 537-541 (1997)
[Publications] Ishiguro A.: "Effects and serum levels of thrombopoietin in a case of chronic thrombocytopenia with achondroplasia." Int.J.Hematol.66. 99-102 (1997)
[Publications] Muraoka K.: "Thrombopoietin-independent effect of interferon-γ on the proliferation of human megakaryocyte progenitors." Brit.J.Haematol.98. 265-273 (1997)
[Publications] Nishijima I.: "Hemopoietic and lymphopoietic responses in human GM-CSF receptor transgenic mice injected with human GM-CSF." Blood. 90. 1031-1038 (1997)
[Publications] Yoshida M.: "Thrombopoietin alone stimulates the early proliferation and survival of human erythroid,myeloid,and multipotential progenitors in serum-free culture." Birt.J.Haematol.98. 254-264 (1997)
[Publications] Umemoto Y.: "Leptin stimulates the proliferation of murine myelocytic and primitive hematopoietic progenitor cells." Blood. 90. 3438-3443 (1997)
[Publications] Akasaka T.: "The role of mel-18,a Mammalian Polycomb group gene,during the IL-7-dependent proliferation of lymphocyte precursors." Immunity. 7. 135-146 (1997)
[Publications] Ebihara Y.: "Synergistic action of Flt3 and gp130 signalings in human hematopoiesis." Blood. 90. 4363-4368 (1997)
[Publications] Tsuji K.: "Role of soluble interleukin-6 receptor with interleukin-6 in human hematopoiesis." Hematology. 2. 143-149 (1997)
[Publications] Suzuki H.: "An increase in peripheral blood progenitor cells in primates by co-administration of recombinant human interleukin 6 and recombinant human granulocyte colony-stimulating factor." Transplantation. 64. 1468-1473 (1997)
[Publications] Bessho M.: "Treatment of the anemia of aplastic anemia patients with recombinant human erythropoietin in combination with granulocyte colony-stimulating factor. a multicenter randomized controlled study." Eur.J.Haematol.58. 265-272 (1997)
[Publications] Suzuki H.: "Early and late events in FcεRI signal transduction in human cultured mast cells." The Journal of Immunology. 159. 5881-5888 (1997)
[Publications] Takahashi T.: "A novel molecular approach to ex vivo hematopoietic expansion with recombinant EGFR-expressing adenovirus vector." Blood. (in press.).
[Publications] Toru H.: "IL-4 promotes the development of tryptase and chymase double positive human mast cells accompanied with cell maturation." Blood. (in press.).
[Publications] Manabe A.: "Characterization of leukemic cells in CD2/CD19 double positive acute lymphoblastic leukemia." Int.J.Hematol.(in press.).
[Publications] Sui X.: "Soluble IL-6 receptor with IL-6 stimulates megakaryopoiesis from human CD34_+ cells through gp130 signaling." Blood. (in press.).
[Publications] Nakahata T.: "Ex vivo expansion of human primitive hemopoietic progenitors." Stem Cells.(in press.).
[Publications] Mukouyama Y.: "Induction of hematopoiesis by oncostatin M in the AGM region." Immunity. (in press.).
[Publications] Nishijima I.: "Human granulocyte-macrophage colony stimulating factor(hGM-CSF)dependent in vitro and in vivo proliferation and differentiation of all hematopoietic progenitor cells in hGM-SCF receptor transgenic mice." Allergy and Clin.Immunol.(in press.).
[Publications] Nishihara M.: "A combination of stem cell factor and granulocyte xolony-stimulating factor enhances proliferation and differentiation of human progenitor cells supported by murine stromal cell line MS-5." Euro.J.Immunol.(in press.).
[Publications] Tsuji K.: "Interferon-γ and human megakaryopoiesis." Leukemia and Lymphoma. (in press.).
[Publications] Tajika K.: "IL-6 and SCF exart different effects on megakaryocyte maturation." Brit.J.Haematol.(in press.).
[Publications] Maruyama K.: "Characterization of peripheral blood progenitor cells mobilized by Nartograstim(N-terminal replaced granulocyte colony-stimulating factor)in normal volunteers." Bone Marrow Transplant.(in press.).
[Publications] Xu M.: "Stimulation of human primitive hematopoiesis by murine AGM-derived stromal cells." Blood. (in press.).
[Publications] Ishiguro A.: "Elevation of serum thrombopoietin precedes thrombocytopoiesis in Kawasaki disease." Thromb.Haemostasis. (in press.).

1997 Fiscal Year Annual Research Report

ヒトG-CSFレセプタートランスジェニックマウスを用いた白血病発症機構の研究

Principal Investigator

中畑 龍俊 東京大学, 医科学研究所, 教授 (20110744)

Research Products

[Publications] Muraoka K.: "Defective response to thrombopoietin and impared expression of c-mpl mRAD in bone marrow cells in congenital amegakaryocytic thrombocytopenia." Brit.J.Haematol.96. 287-292 (1997)

[Publications] Eguchi M.: "Morphological and cytochemical changes in human mast cells during culture." Med.Electron Microsc.30. 25-30 (1997)

[Publications] Toru H.: "Interleukin-4 induces homotypic aggregation of human mast cells by promoting LFA-1/ICAM-1 adhesion molecules." Blood. 89. 3296-3302 (1997)

[Publications] Yanagida M.: "Effect of a chymotrypsin-like inhibitor,TPCK,on histamine release from cultured human mast cells." J.Pharm.Pharmth.49. 537-541 (1997)

[Publications] Ishiguro A.: "Effects and serum levels of thrombopoietin in a case of chronic thrombocytopenia with achondroplasia." Int.J.Hematol.66. 99-102 (1997)

[Publications] Muraoka K.: "Thrombopoietin-independent effect of interferon-γ on the proliferation of human megakaryocyte progenitors." Brit.J.Haematol.98. 265-273 (1997)

[Publications] Nishijima I.: "Hemopoietic and lymphopoietic responses in human GM-CSF receptor transgenic mice injected with human GM-CSF." Blood. 90. 1031-1038 (1997)

[Publications] Yoshida M.: "Thrombopoietin alone stimulates the early proliferation and survival of human erythroid,myeloid,and multipotential progenitors in serum-free culture." Birt.J.Haematol.98. 254-264 (1997)

[Publications] Umemoto Y.: "Leptin stimulates the proliferation of murine myelocytic and primitive hematopoietic progenitor cells." Blood. 90. 3438-3443 (1997)

[Publications] Akasaka T.: "The role of mel-18,a Mammalian Polycomb group gene,during the IL-7-dependent proliferation of lymphocyte precursors." Immunity. 7. 135-146 (1997)

[Publications] Ebihara Y.: "Synergistic action of Flt3 and gp130 signalings in human hematopoiesis." Blood. 90. 4363-4368 (1997)

[Publications] Tsuji K.: "Role of soluble interleukin-6 receptor with interleukin-6 in human hematopoiesis." Hematology. 2. 143-149 (1997)

[Publications] Suzuki H.: "An increase in peripheral blood progenitor cells in primates by co-administration of recombinant human interleukin 6 and recombinant human granulocyte colony-stimulating factor." Transplantation. 64. 1468-1473 (1997)

[Publications] Bessho M.: "Treatment of the anemia of aplastic anemia patients with recombinant human erythropoietin in combination with granulocyte colony-stimulating factor. a multicenter randomized controlled study." Eur.J.Haematol.58. 265-272 (1997)

[Publications] Suzuki H.: "Early and late events in FcεRI signal transduction in human cultured mast cells." The Journal of Immunology. 159. 5881-5888 (1997)

[Publications] Takahashi T.: "A novel molecular approach to ex vivo hematopoietic expansion with recombinant EGFR-expressing adenovirus vector." Blood. (in press.).

[Publications] Toru H.: "IL-4 promotes the development of tryptase and chymase double positive human mast cells accompanied with cell maturation." Blood. (in press.).

[Publications] Manabe A.: "Characterization of leukemic cells in CD2/CD19 double positive acute lymphoblastic leukemia." Int.J.Hematol.(in press.).

[Publications] Sui X.: "Soluble IL-6 receptor with IL-6 stimulates megakaryopoiesis from human CD34_+ cells through gp130 signaling." Blood. (in press.).

[Publications] Nakahata T.: "Ex vivo expansion of human primitive hemopoietic progenitors." Stem Cells.(in press.).

[Publications] Mukouyama Y.: "Induction of hematopoiesis by oncostatin M in the AGM region." Immunity. (in press.).

[Publications] Nishijima I.: "Human granulocyte-macrophage colony stimulating factor(hGM-CSF)dependent in vitro and in vivo proliferation and differentiation of all hematopoietic progenitor cells in hGM-SCF receptor transgenic mice." Allergy and Clin.Immunol.(in press.).

[Publications] Nishihara M.: "A combination of stem cell factor and granulocyte xolony-stimulating factor enhances proliferation and differentiation of human progenitor cells supported by murine stromal cell line MS-5." Euro.J.Immunol.(in press.).

[Publications] Tsuji K.: "Interferon-γ and human megakaryopoiesis." Leukemia and Lymphoma. (in press.).

[Publications] Tajika K.: "IL-6 and SCF exart different effects on megakaryocyte maturation." Brit.J.Haematol.(in press.).

[Publications] Maruyama K.: "Characterization of peripheral blood progenitor cells mobilized by Nartograstim(N-terminal replaced granulocyte colony-stimulating factor)in normal volunteers." Bone Marrow Transplant.(in press.).

[Publications] Xu M.: "Stimulation of human primitive hematopoiesis by murine AGM-derived stromal cells." Blood. (in press.).

[Publications] Ishiguro A.: "Elevation of serum thrombopoietin precedes thrombocytopoiesis in Kawasaki disease." Thromb.Haemostasis. (in press.).

中畑龍俊東京大学, 医科学研究所, 教授 (20110744)