Studies on causative gene defects in primary neutrophil abnormalities with a purpose of applying it to the gene therapy

原发性中性粒细胞异常致病基因缺陷的研究及其应用于基因治疗的目的

• 批准号: 06454299
• 负责人: KOMIYAMA Atsushi
• 金额: $ 3.78万
• 依托单位: Shinshu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454299/
• 关键词: neutrophil; neutrophil functtion; immunodeficiency; neutropenia; neutrophil dysfunction; neutrophil-specific granule deficiency; G-CSF receptor; bactericidal proteins; 好中球機能不全症; デフェンシン; ラクトフェリン; コロニー刺激因子レセプター

The present studies were conducted to analyze the induction of neutrophil functions and its signal transduction and to determine the causative gene defects in primary neutrophil abnormalities with a purpose of applying it to the gene therapy.1. Analysis of induction of neutrophil functions and its signal transduction(1) Neutrophil functions were induced via CD11b, an adhesion molecule, which plays a role in regulating the functions by the interaction between neutrophils and endothelial cells.(2) The tyrosine kinase was demonstrated to be a specific signaling pathway for FMLP-induced chemotaxis.2. Analysis of causative gene defects in primary neutrophil abnormalities(1) Congenital neutropenia (kostmann type) : It was possible to get neutrophils enough to use for the present studies by culturing the bone marrow cells in two patients. The G-CSF receptor was not decreased in the number, and its mRNA expression was normal.(2) Congenital neutrophil-specific granule deficiency : There was no or reduced expression of mRNA for not only defensins but also lactoferrin and collagenase. Southern blot analysis of these genomic DNAs failed to demonstrate abnormal band by use of several restriction enzymes. Studies are now in progress to analyze the flanking regions and promoter of the genes.

本研究旨在分析中性粒细胞功能的诱导及其信号转导，并确定原发性中性粒细胞异常的致病基因缺陷，以期将其应用于基因治疗.中性粒细胞功能的诱导及其信号转导分析（1）中性粒细胞功能是通过粘附分子CD 11b诱导的，CD 11b通过中性粒细胞与内皮细胞的相互作用发挥调节作用。(2)酪氨酸激酶被证明是FMLP诱导趋化性的特异性信号通路.原发性中性粒细胞异常致病基因缺陷的分析（1）先天性中性粒细胞减少症（Kostmann型）：通过培养2例患者的骨髓细胞，有可能获得足够的中性粒细胞用于本研究。G-CSF受体数量未减少，其mRNA表达正常。(2)先天性嗜中性粒细胞特异性颗粒缺乏：不仅防御素，而且乳铁蛋白和胶原酶的mRNA表达没有或减少。用几种限制性内切酶对这些基因组DNA进行Southern印迹分析，均未发现异常条带。目前正在对这些基因的侧翼区和启动子进行分析。

项目成果

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Yasui,K.: "Signal transduction pathway in human PMN for chemotaxis induced by a chemotactic factor." J.Immunol.152. 5922-5929 (1994)

Yasui,K.：“人中性粒细胞中趋化因子诱导趋化作用的信号转导途径。”

安井耕三: "新生児の好中球:機能と生理" 日本産婦人科・新生児血液学会雑誌. 5. 48-59 (1995)

Kozo Yasui：“新生儿中性粒细胞：功能和生理学”日本妇产科和新生儿血液学会杂志 5. 48-59 (1995)。

小宮山 淳: "血液病学" 三輪史朗他(編),文光堂, 1813 (1995)

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Tamura,A.: "A marked decrease in defensin mRNA in the only case of congenital neutrophil‐specific granule deficiency." International Journal of Hematology. 59. 137-142 (1994)

Tamura, A.：“在唯一的先天性中性粒细胞特异性颗粒缺陷病例中，防御素 mRNA 显着减少。”《国际血液学杂志》59. 137-142 (1994)