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造血幹細胞・リンパ球前駆細胞の増幅分化制御とその免疫監視機構再生への応用

造血干细胞和淋巴祖细胞扩增和分化的控制及其在免疫监视系统再生中的应用

基本信息

批准号：
16043213
负责人：
高木智
金额：
$ 3.58万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
2004
资助国家：
日本
起止时间：
2004 至无数据
项目状态：
已结题

项目摘要

1)造血幹細胞・リンパ球前駆細胞の増幅を制御する分子機構の解明:Lnk欠損により機能的な骨髄造血幹細胞が著増すること、一部の造血幹細胞においては細胞あたりの造血系再構築能も亢進していることを個々の造血幹細胞の競合的骨髄再構築能を個別に検討することにより明らかにした。CD34陰性Kit陽性Sca-1陰性の造血幹細胞における遺伝子発現プロファイルの変化についてジーンチップによる検討を推進した。2)骨髄内における造血幹細胞のホーミング機構の解析:Lnkを過剰発現させた線維芽細胞株ではアクチン細胞骨格に変化が生じ、細胞分裂や細胞遊走が阻害されることを明らかにした。分化抗原陰性の造血前駆細胞を正常及びLnk欠損マウスより精製し異なる蛍光色素で標識して静注したところ、Lnk欠損前駆細胞の骨髄への分布が亢進している可能性が示唆された。3)造血幹細胞、リンパ球前駆細胞の分化増殖制御に向けての基礎技術開発:Lnk阻害法を開発し、免疫監視機構の再生能亢進への応用の基盤整備を目指した。種々のLnk変異体をc-Kit依存性に増殖する細胞株に発現させ機能ドメインを同定した。増殖抑制にはSH2ドメインが必須であり、SH2変異体はドミナントネガティブ(DN)変異体として機能することがわかった。SH2変異に加えてPHドメイン欠損とC末端領域欠損を組み合わせることで、より効果的なDn-Lnk変異体を作出した。得られたDN-Lnk変異体をレトロウィルスベクターによりマウス造血前駆細胞に感染導入し骨髄移植実験系で効果を検討した結果、DN-Lnk変異体を導入した前駆細胞の造血能亢進が観察された。さらに、DN-Lnk変異体のプラスミドDNAによる一過性発現によっても造血前駆細胞の生着能が亢進すること、非骨髄破壊的移植条件下でも免疫不全マウスの免疫監視機構を効率よく再建できることがわかった。

1) the molecular mechanism for the regulation and control of pre-globular hematopoietic cells showed that the bone marrow hematopoietic cells of Lnk deficient cells were involved in this disease, and that the reassortment of the hematopoietic system of one hematopoietic cell cycle could enhance the activation of two different hematopoietic cell complexes. CD34, Kit, Sca-1, hematopoiesis, hematopoiesis, hematopoi 2) Intraosseous hematopoietic progenitor cell cytological analysis: Lnk assay showed that the cell line, cell line, cellular lattice, mitotic cell cycle, cell cycle Differentiation antigens prehematopoietic progenitor cells were normal and Lnk deficient hematopoietic prehematopoietic progenitor cells were normal and Lnk deficient prehematopoietic progenitor cells were identified by intravenous injection, bone marrow distribution, and the possibility of hyperactivity. 3) the differentiation of hematopoietic progenitor cells and preglobular progenitor cells and the development of basic techniques: the Lnk blocking method is used, and the regeneration energy of immune monitoring institutions is enhanced. The c-Kit-dependent colonization of all kinds of Lnk strains showed that the mechanism was the same as that of the control group. Colonization inhibition is necessary for the SH2 population. The SH2 system must be activated, and the DN system must be activated. The mechanism will not be affected. The SH2 system adds the PH information to make sure that the C-terminal domain is not available and that the Dn-Lnk system of the result is not available. The infection of prehematopoietic progenitor cells was detected in the bone transplant system. The results showed that the hematopoietic activity of DN-Lnk prehematopoietic progenitor cells was increased. The results showed that the hematopoietic activity of pre-hematopoietic progenitor cells was significantly higher than that of normal bone transplantation. the results showed that the hematopoietic activity of prohematopoietic cells was increased after bone transplantation. It was found that the pre-hematopoietic progenitor cells were born under the condition of transplantation under the condition of non-bone fracture under the condition of transplantation. the rate of immune monitoring institutions of immune insufficiency was significantly higher than that of normal graft. as soon as it was found that the prehematopoietic progenitor cells were born under the condition of transplantation under the condition of transplantation without bone fracture, the rate of immune monitoring institutions and the rate of immune monitoring institutions were reestablished.