Ex vivo expansion of mature neutrophils from hematopoietic progenitor cells and cytokine-mediated signal transduction

造血祖细胞成熟中性粒细胞的离体扩增和细胞因子介导的信号转导

基本信息

批准号：
14570992
负责人：
HINO Masayuki
金额：
$ 1.86万
依托单位：
Osaka City University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570992/
关键词：
hematopoietic progenitor cell neutrophil G-CSF TPO MAPkinase STAT IAP JAK サイトカインシグナル伝達アポトーシス JAK-STAT

项目摘要

In CD34 positive hematopoietic progenitor cells, stem cell factor(SCF), granulocyte colony-stimulating factor(G-CSF), thrombopoietin(TPO), granulocyte-macrophage colony-stimulating factor(GM-CSF), interleukin(IL)-3, and tumor necrosis factor(TNF)-α mediated phosphorylation of extracellular-signal regulated kinase(ERK), IL-1 and TNF-α mediated phosphorylation of p38 MAP kinase, TNF-α mediated phosphorylation of c-Jun N-terminal kinase (JNK). Inhibitors of p38 MAP kinase inhibited expression of CD11b and CD15 induced by IL-1b. IL-6/sIL-6R,G-CSF,TPO,GM-CSF,IL-3,IFN-α, and IFN-γ mediated phosphorylation of signal transducer and activator of transcription(STAT)3-Y,SCF,IL-6/sIL-6R,G-CSF,TPO,GM-CSF,IL-3, and TNF-α mediated phosphorylation of STST3-S,G-CSF,TPO,GM-CSF, IL-3, and IFN-a mediated phosphorylation of STST5a,G-CSF,GM-CSF,IL-3,IFN-α, and IFN-γ mediated phosphorylation of STAT5b, and IFN-α and IFN-γ mediated phosphorylation of STAT 1.Mature human neutrophils with normal functions (chemotaxis, phagocytosis and superoxide release) were ex vivo expanded from haematopoietic progenitor cells as the following procedure. CD34 positive haematopoietic progenitor cells were cultivated with SCF,IL-3,GM-CSF, and G-CSF for 7 days, and thereafter non-adherent cells were further cultivated for 7 days with G-CSF alone. TPO stimulated ex vivo expansion of mature neutrophils in the early stages of differentiation. Human neutrophils were expressed members of the inhibitor of apoptosis (LAP) family (cIAP1,cIAP2, and X-linked LAP). G-CSF selectively up-regulated c-IAP2 expression. A specific inhibitor of Janus kinase 2(JAK2) inhibited G-CSF-induced up-regulation of cIAP2, phosphorylation of STAT3, and the antiapoptotic effects. In monocyte, G-CSF inhibited LPS-induced TNF-α production and activated STST3. These effects were prevented by an inhibitor of JAK2.

在CD34阳性造血祖细胞中，干细胞因子（SCF），粒细胞菌落刺激因子（G-CSF）（G-CSF），血小板蛋白（TPO），粒细胞 - 巨噬细胞刺激性刺激因子（GM-CSF），ILSIRDIAN-3，IL）-3，和TUMOR NECRISITIAL NECRIANT-FLAPHORY necrosis NFSROSIS（TNF）（TNF）（TNF）（TNF）。 p38 MAP激酶的细胞外信号调节激酶（ERK），IL-1和TNF-α介导的磷酸化，TNF-α介导的C-JUN N末端激酶（JNK）的介导的磷酸化。 p38 MAP激酶的抑制剂抑制了IL-1B诱导的CD11b和CD15的表达。 IL-6/SIL-6R，G-CSF，TPO，GM-CSF，IL-3，IFN-α和IFN-γ介导的信号转录器和转录激活因子（STAT）3-Y，SCF，IL-6/SIL-6/SIL-6/SIL-6R，G-CSF，TPO，GM-CSF，GM-CSF，GM-CSF，gm-csf，gm-csf，gm-csf，gm-csf，gm-csf，gm-csf，gm-csf，and csf，和STST3-S，G-CSF，TPO，GM-CSF，IL-3和IFN-A介导的STST5A，G-CSF，GM-CSF，GM-CSF，IL-3，IFN-α，IFN-α和IFN-γ介导的STAT5B和IFN-α和IFN-γ介导的人类磷酸化的磷酸化的磷酸化和IFN-γ介导的磷酸化。（趋化性，吞噬作用和超氧化物释放）是从造血祖细胞扩展的，作为以下过程。 CD34阳性造血祖细胞用SCF，IL-3，GM-CSF和G-CSF培养7天，然后单独使用G-CSF进一步培养非粘附细胞7天。在分化的早期阶段，TPO刺激了成熟中性粒细胞的离体扩张。人类嗜中性粒细胞表达了凋亡（LAP）家族抑制剂（CIAP1，CIAP2和X连接圈）的成员。 G-CSF选择性上调的C-IAP2表达。 Janus激酶2的特定抑制剂（JAK2）抑制了G-CSF诱导的CIAP2上调，STAT3的磷酸化和抗凋亡作用。在单核细胞中，G-CSF抑制了LPS诱导的TNF-α产生并活化的STST3。这些作用是由JAK2抑制剂阻止的。