1996 Fiscal Year Annual Research Report

分化抑制による造血幹細胞の体外増幅

Research Project

Project/Area Number	08261210
Research Institution	Kyushu University
Principal Investigator	岡村精一九州大学, 医学部, 講師 (20136435)
Co-Investigator(Kenkyū-buntansha)	浅野嘉延九州大学, 医学部, 助手 (60271110) 権藤久司九州大学, 医学部, 助手 (10253428) 岡村孝九州大学, 医学部, 助手 (30136436)
Keywords	可溶性G-CSFレセプター / G-CSF / 造血幹細胞 / 先天性好中球減少症 / 急性骨髄性白血病 / 遺伝子発現 / スプライシング / セルソーター
Research Abstract	G-CSFは幼若造血幹細胞にも働き,分化成熟も強力に刺激する.このG-CSFは骨髄細胞からも内因性に産生される.G-CSFの作用を可溶性G-CSFレセプターでブロックする事により,分化成熟抑制して増殖の方向に比較的誘導できる可能性を考え,まず可溶性G-CSFレセプターに関し検討した. ヒト血清をビオチン標識したG-CSFと孵置した後にnative PAGEで泳動し,ニトロセルロース膜に移したものとperoxidase(PO)標識streptavidinと反応させPOを発色させた.95kDと105kDにバンドが検出され,G-CSFの分子量約20kDを減じると,75kDと85kDのG-CSFと結合する血清蛋白の存在が考えられた.さらに,抗G-CSFレセプター抗体を用いた実験でもほぼ同じ大きさに2つのバンドを認めた. ヒト好中球系細胞の成熟段階における,可溶性G-CSFレセプターの発現状況をRT-PCR法で検索した.ヒト骨髄細胞をセルソーターで成熟段階にしたがい,分離してG-CSFレセプターの膜貫通部を挟むプライマーを用いてRT-PCRを行うと,完全長型と膜貫通部を欠いた可溶性型G-CSFレセプター由来の2つのバンドが検出され,可溶性G-CSFレセプター由来の短いバンドは成熟段階が進行するに従いその割合が増加していた. G-CSFレセプターの体細胞遺伝子は17のエクソンからなり,全体ではないが第15エクソンの大部分が膜貫通部をコードしている.先天性好中球減少症患者より第15エクソン全体が欠失しているクローンを得た.これらはヒト骨髄性白血病細胞株の多くでも検出された.このcDNAを発現ベクターに入れCHO-K1細胞に遺伝子導入した. さらに,マウス可溶性G-CSFレセプターを試験管内で作用させ,正常G-CSF作用を抑制すること,及びヒト骨髄性白血病細胞の増殖を抑制することを見いだした.

Research Products
(17 results)

All Other

All Publications (17 results)

[Publications] Takenaka K.: "Generation of human natural killer cells from peripheral blood CD34+ cells mobilized by granulocyte colony-stimulating factor" British Journal of Haematology. 92(4). 788-794 (1996)
[Publications] Kuroiwa M.: "Effects of granulocyte colony-stimulating factor on the hemostatic system in healthy volunteers" International Journal of Hematology. 63(4). 311-316 (1996)
[Publications] Asano Y.: "Effect of(-)-epigallocatechin gallate on leukemic blast cells from patients with acute myeloblastic leukemia" Life Sciences. 60(2). 135-142 (1997)
[Publications] Miyamoto T.: "Persistence of multipotent progenitors expressing AML1/ETO transcripts in long-term remission patients with t(8;21)acute myelogenous leukemia" Blood. 87(11). 4789-4796 (1996)
[Publications] Kanaji T.: "A sequence analysis of von Willebrand factor mRNA,gene,and pseudogene in two patients with von Willebrand disease type 2B,and an investigation of gene conversion in its gene" International Journal of Hematology. 64(1). 53-59 (1996)
[Publications] Harada M.: "G-CSF-induced mobilization of peripheral blood stem cells from healthy adults for allogeneic transplantation" Journal of Hematotherapy. 5(1). 63-71 (1996)
[Publications] Miyamoto T.: "Serum concentration of the soluble interleukin-2 receptor for monitoring acute graft-versus-host disease" Bone Marrow Transplantation. 17(2). 185-190 (1996)
[Publications] Harada M.: "Granulocyte colony-stimulating factor-induced mobilization of peripheral blood stem cells for autologous and allogeneic transplantation" Cancer Chemotherapy Pharmacology. 38Suppl. 115-119 (1996)
[Publications] Tsutsui,S.: "Prognostic significance of immunological parameters in patients with esophageal cancer. In vitro study of deoxyspergualin on function of lymphocytes and bone marrow cells from healthy volunteers" Hepato-Gastroenteology. 43. 501-509 (1996)
[Publications] Matsunaga T.: "Rotenone,a mitochondrial NADH dehydrogenase inhibitor,induces cell surface expression of CD 13 and CD38 and apoptosis in HL-60 cells" Leukemia Lymphoma. 20. 487-494 (1996)
[Publications] Chen,F.: "Reversible primary hypothyroidism with blocking or stimulating type TSH binding inhibitor immunoglobulin following recombinant interferon-a therapy in patients with pre-existing thyroid disorders" Clinical Endocrinology. 45. 207-214 (1996)
[Publications] Takeshita M.: "Angiocentric immunoproliferative lesions of the lymph node" American Journal of Clinical Pathology. 106(1). 69-77 (1996)
[Publications] 岡村精一: "G-CSFレセプターを介する増殖・分化のシグナル伝達" 医学のあゆみ. (印刷中).
[Publications] 岡村精一: "造血幹細胞からの顆粒球系細胞の分化" Mebio. (印刷中).
[Publications] Asano Y.: "Analysis of G-CSF receptor in various hematological disorders.Approaches to Diseases(Niho Y.ed)" Kyushu University Press.,Fukuoka, 12 (1996)
[Publications] 岡村精一: "G-CSFレセプターのシグナル伝達機構とその異常 Annal Rwview 血液97" 中外医学社,東京, 16 (1997)
[Publications] 岡村精一: "G-CSF受容体異常キーワード1997-'98血液" 先端医学社(印刷中),

1996 Fiscal Year Annual Research Report

分化抑制による造血幹細胞の体外増幅

Principal Investigator

岡村 精一 九州大学, 医学部, 講師 (20136435)

Research Products

[Publications] Takenaka K.: "Generation of human natural killer cells from peripheral blood CD34+ cells mobilized by granulocyte colony-stimulating factor" British Journal of Haematology. 92(4). 788-794 (1996)

[Publications] Kuroiwa M.: "Effects of granulocyte colony-stimulating factor on the hemostatic system in healthy volunteers" International Journal of Hematology. 63(4). 311-316 (1996)

[Publications] Asano Y.: "Effect of(-)-epigallocatechin gallate on leukemic blast cells from patients with acute myeloblastic leukemia" Life Sciences. 60(2). 135-142 (1997)

[Publications] Miyamoto T.: "Persistence of multipotent progenitors expressing AML1/ETO transcripts in long-term remission patients with t(8;21)acute myelogenous leukemia" Blood. 87(11). 4789-4796 (1996)

[Publications] Kanaji T.: "A sequence analysis of von Willebrand factor mRNA,gene,and pseudogene in two patients with von Willebrand disease type 2B,and an investigation of gene conversion in its gene" International Journal of Hematology. 64(1). 53-59 (1996)

[Publications] Harada M.: "G-CSF-induced mobilization of peripheral blood stem cells from healthy adults for allogeneic transplantation" Journal of Hematotherapy. 5(1). 63-71 (1996)

[Publications] Miyamoto T.: "Serum concentration of the soluble interleukin-2 receptor for monitoring acute graft-versus-host disease" Bone Marrow Transplantation. 17(2). 185-190 (1996)

[Publications] Harada M.: "Granulocyte colony-stimulating factor-induced mobilization of peripheral blood stem cells for autologous and allogeneic transplantation" Cancer Chemotherapy Pharmacology. 38Suppl. 115-119 (1996)

[Publications] Tsutsui,S.: "Prognostic significance of immunological parameters in patients with esophageal cancer. In vitro study of deoxyspergualin on function of lymphocytes and bone marrow cells from healthy volunteers" Hepato-Gastroenteology. 43. 501-509 (1996)

[Publications] Matsunaga T.: "Rotenone,a mitochondrial NADH dehydrogenase inhibitor,induces cell surface expression of CD 13 and CD38 and apoptosis in HL-60 cells" Leukemia Lymphoma. 20. 487-494 (1996)

[Publications] Chen,F.: "Reversible primary hypothyroidism with blocking or stimulating type TSH binding inhibitor immunoglobulin following recombinant interferon-a therapy in patients with pre-existing thyroid disorders" Clinical Endocrinology. 45. 207-214 (1996)

[Publications] Takeshita M.: "Angiocentric immunoproliferative lesions of the lymph node" American Journal of Clinical Pathology. 106(1). 69-77 (1996)

[Publications] 岡村精一: "G-CSFレセプターを介する増殖・分化のシグナル伝達" 医学のあゆみ. (印刷中).

[Publications] 岡村精一: "造血幹細胞からの顆粒球系細胞の分化" Mebio. (印刷中).

[Publications] Asano Y.: "Analysis of G-CSF receptor in various hematological disorders.Approaches to Diseases(Niho Y.ed)" Kyushu University Press.,Fukuoka, 12 (1996)

[Publications] 岡村精一: "G-CSFレセプターのシグナル伝達機構とその異常 Annal Rwview 血液97" 中外医学社,東京, 16 (1997)

[Publications] 岡村精一: "G-CSF受容体異常 キーワード1997-'98血液" 先端医学社(印刷中),

岡村精一九州大学, 医学部, 講師 (20136435)

[Publications] 岡村精一: "G-CSF受容体異常キーワード1997-'98血液" 先端医学社(印刷中),