Cytomegalovirus infection stimulates expression of monocyte-associated mediator genes.
Cytomegalovirus infection stimulates expression of monocyte-associated mediator genes.
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DOI：
10.4049/jimmunol.143.10.3343
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发表时间：
1989-11
影响因子：
4.4
通讯作者：
L. Dudding;S. Haskill;B. D. Clark;P. Auron;S. Sporn;E. Huang
中科院分区：
文献类型：
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作者：
L. Dudding;S. Haskill;B. D. Clark;P. Auron;S. Sporn;E. Huang
单核细胞和组织巨噬细胞在宿主防御病毒感染中起重要作用，在人巨细胞病毒（HCMV）和HIV的情况下，也可能是潜伏性疾病的储存库。由于这些细胞也可以通过分泌炎症介质对大多数感染迅速作出反应，我们有兴趣确定HCMV感染是否对巨噬细胞细胞因子产生直接激活作用。为此，我们主要研究了HCMV感染对外周血单核细胞和早幼粒细胞系ML-3中IL-1 β-mRNA表达以及早幼粒细胞系ML-3中炎症反应基因TNF-α、MAD-9、MAD-6和MAD-2的影响。ML-3细胞在诱导分化前暴露于病毒对介体基因表达的影响不大。然而，通过用佛波醇酯PMA预处理ML-3细胞，随后进行HCMV攻击来诱导巨噬细胞表型，导致IL-1 β、TNF-α、MAD-9和CSF-1的表达时间大大延长，但MAD-6和MAD-2的表达时间没有延长。组成型表达的基因，如溶菌酶和肌动蛋白没有类似的调制。RNA斑点杂交和原位杂交研究均表明，人外周血单核细胞感染HCMV可导致IL-1 β mRNA持续表达长达96小时，这与模拟感染或LPS刺激的单核细胞形成明显对比。对ML-3细胞中IL-1 β蛋白的细胞内水平的流式细胞术分析表明，不仅在感染的细胞中产生更多的蛋白，而且大多数细胞都有反应。通过Western印迹分析证实单核细胞中IL-1 β的细胞内形式的水平提高。使用IL-1 β-CAT嵌合质粒与编码HCMV立即早期基因区产物的质粒一起进行共转染实验。在转染前已诱导分化的ML-3细胞中观察到IL-1 β基因被立即早期基因的区域2反式激活。在转染前未分化的ML-3细胞中未观察到IL-1 β启动子活性的刺激。总之，HCMV感染，虽然不导致生产性感染，但可能有助于通过增强单核细胞炎症介质基因表达，随后刺激蛋白质合成的感染的病理。
Monocytes and tissue macrophages play important roles in host defense against virus infections and, in the case of human cytomegalovirus (HCMV) and HIV, may also be the reservoir for latent disease. Because these cells can also rapidly respond to most infections by secretion of inflammatory mediators, we were interested in determining if HCMV infection could have a direct activating effect on macrophage cytokine production. To do this, we primarily investigated the influence of HCMV infection on IL-1 beta-mRNA expression in peripheral blood monocytes and the promyelocytic cell line, ML-3 as well as the inflammatory response genes TNF-alpha, MAD-9, MAD-6, and MAD-2 in the promyelocytic ML-3 cell line. Exposure of ML-3 cells to the virus prior to induction of differentiation had little influence on mediator gene expression. However, induction of the macrophage phenotype by pretreatment of ML-3 cells with the phorbol ester, PMA, followed by HCMV challenge, resulted in a greatly extended period of expression of IL-1 beta, TNF-alpha, MAD-9, and CSF-1 but not MAD-6 and MAD-2. Constitutively expressed genes such as lysozyme and actin were not similarly modulated. Both RNA dot-blot and in situ hybridization studies demonstrated that infection of human peripheral blood monocytes with HCMV leads to sustained expression of IL-1 beta mRNA for up to 96 h, which contrasted markedly with mock-infected or LPS-stimulated monocytes. Flow cytometric analysis of the intracellular levels of IL-1 beta protein in ML-3 cells indicated that not only was there more protein produced in infected cells, but that the majority of the cells had responded. Enhanced levels of the intracellular form of IL-1 beta in monocytes was confirmed by Western blot analysis. Cotransfection experiments were performed using IL-1 beta-CAT chimeric plasmids together with plasmids encoding HCMV-immediate-early gene region products. Transactivation of the IL-1 beta gene by region 2 of the immediate-early gene was observed in ML-3 cells that had been induced to differentiate prior to transfection. No stimulation of IL-1 beta promoter activity was observed in ML-3 cells that were undifferentiated prior to transfection. In summary, HCMV infection, although not leading to productive infection, nonetheless may contribute to the pathology of the infection through enhancement of monocyte inflammatory mediator gene expression with subsequent stimulation of protein synthesis.