Molecular mechanisms of IL-8 gene expression by the immediate early gene products of cytomegelovirus

巨细胞病毒早期基因产物表达IL-8基因的分子机制

• 批准号: 09670320
• 负责人: MURAYAMA Tsugiya
• 金额: $ 1.98万
• 依托单位: Kagoshima University (1999)Kanazawa Medical University (1997-1998)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670320/
• 关键词: Cytomegalovirus; IL-8; U373MG cell; luciferase reporter gene assay; NF-kB; AP-1; ゲルシフトアッセイ; U373MG細胞; 免疫染色; 遺伝子導入; THP-1細胞

We previously observed that human cytomegalovires (CMV) infection induced a massive production of an interleukin-8 (IL-8) in a THP-1 cell. Hence, we examined the effect of CMV immediate early (IE) gene products on IL-8 production by the human astrocytoma cell line, U373MG. Transiently or stably transfection with CMV IE1 but not with CMV IE2 gene significantly augmented IL-8 protein secretion and IL-8 mRNA expression, compared with transfection with a control vector. Moreover, luciferase activity was enhanced in U373MG cells that were co-transfected with CMV IE1 and a chimeric firefly luciferase reporter genes driven by the transcriptional regulatory region of the human IL-8 gene. Furthermore, IE1 gene-mediated enhancement of luciferase activities was abolished by the introduction of the mutation into AP-1 or NF-kB factor binding elements in the regulatory region of the IL-8 gene, as previously observed on CMV infected cells. Finally, Western blotting as well as an immunohistochemical analysis demonstrated that CMV IE1 gene products increased the amount of p65 of NF-kB complexes translocated into the nuclei. Collectively, CMV IE1 gene expression may be sufficient to activates AP-1 and NF-kB, resulting in IL-8 gene expressions.

我们以前观察到，人巨细胞病毒（CMV）感染诱导THP-1细胞中大量产生白细胞介素-8（IL-8）。因此，我们研究了CMV立即早期（IE）基因产物对人星形细胞瘤细胞系U373 MG产生IL-8的影响。瞬时或稳定转染CMV IE 1基因而不转染CMV IE 2基因显著增加IL-8蛋白分泌和IL-8 mRNA表达。此外，荧光素酶活性在U373 MG细胞中增强，所述U373 MG细胞用CMV IE 1和由人IL-8基因的转录调控区驱动的嵌合萤火虫荧光素酶报告基因共转染。此外，IE 1基因介导的荧光素酶活性的增强被废除的突变引入到AP-1或NF-κ B因子结合元件的IL-8基因的调控区，如先前观察到的CMV感染的细胞。最后，Western印迹以及免疫组化分析表明，CMV IE 1基因产物增加了NF-κ B复合物的p65易位到细胞核中的量。总的来说，CMV IE 1基因表达可能足以激活AP-1和NF-κ B，导致IL-8基因表达。

项目成果

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T. Murauyama: "Potential involvement of interleukin-8 (IL-8) in the pathogenesis of human cytomegalovirus infection"J. Leukocyte Biol.. 64. 62-67 (1998)

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K.S.A.Khabar,et al.: "The α chemokine,interleukin-8,inhibits the....."J.Exp.Med.. 186. 1077-1085 (1997)

K.S.A.Khabar 等人：“α 趋化因子，白细胞介素 8，抑制......”J.Exp.Med.. 186. 1077-1085 (1997)

T.Murayama,et al.: "Potential involvement of IL-8 in the pathogenesis of....."J.Leukoc.Biol.. 64. 62-67 (1998)

T.Murayama 等人：“IL-8 可能参与……的发病机制”J.Leukoc.Biol.. 64. 62-67 (1998)