Increased production of interleukin-8 in primary human monocytes and in human epithelial and endothelial cell lines after dengue virus challenge
Increased production of interleukin-8 in primary human monocytes and in human epithelial and endothelial cell lines after dengue virus challenge
复制标题
DOI：
10.1128/jvi.76.11.5588-5597.2002
复制
发表时间：
2002-06-01
影响因子：
5.4
通讯作者：
Rothman, AL
中科院分区：
文献类型：
--
作者：
Bosch, I;Xhaja, K;Rothman, AL
更严重的登革病毒感染形式是登革出血热，其特点是血浆渗漏和止血紊乱。由于观察到有更严重疾病表现的患者血清中IL-8水平升高，一项研究开始观察登革病毒体外感染对促炎细胞因子分泌和表达的影响。用酶联免疫吸附试验检测了登革2型病毒(D2V)新几内亚C株(NGC)感染人单核细胞培养上清液中IL-8的水平。此外，逆转录聚合酶链式反应也扩增了该基因的表达。在IL-6、IL-1β、IL-8和肿瘤坏死因子α等致炎细胞因子及其mRNAs的检测中，IL-8在D2V感染后变化最大。同样，293T(人上皮细胞系)和ECV304(内皮细胞系)两个细胞系对D2V NGC是允许的，并通过增加IL-8的合成来应对感染。核因子-kappaB和核因子IL-6是IL-8表达的主要介质。我们对IL-8在ECV304和293T细胞中的转录调控进行了研究，发现IL-8基因的表达诱导只在ECV304细胞中参与了NF-kappaB的激活(P=0.001)，在较小的程度上也只参与了NFIL-6的激活。接下来，我们通过染色质免疫沉淀程序在体内观察到D2V感染后与IL-8启动子结合的核心组蛋白的乙酰化。由感染的单核细胞产生的IL-8，以及可能由内皮细胞或其他上皮细胞产生的IL-8，除了被核因子-kappaB激活外，还与与IL-8启动子结合的组蛋白的高乙酰化有关。我们推测，在这项体外研究中观察到的IL-8合成的总体增加可能在登革出血热和登革休克综合征中出现的血浆渗漏的发病机制中起作用。
The more severe form of dengue virus infection, dengue hemorrhagic fever, is characterized by plasma leakage and derangements in hemostasis. As elevated interleukin-8 (IL-8) levels have been observed in sera from patients with more severe disease manifestations, a study was initiated to look at the effect of dengue virus infection in vitro on proinflammatory cytokine secretion and expression. A significant increase in IL-8 levels in the culture supernatant of primary human monocytes infected with dengue 2 virus (D2V) New Guinea C (NGC) was found by enzyme-linked immunosorbent assay. Additionally, by reverse transcriptase PCR, the mRNA was also augmented. Among the proinflammatory cytokines and their mRNAs measured (IL-6, IL-1beta, IL-8, and tumor necrosis factor alpha), IL-8 showed the greatest change following D2V infection. Similarly, two cell lines, 293T (a human epithelial cell line) and ECV304 (an endothelial cell line), were permissive to D2V NGC and responded to the infection by increasing the synthesis of IL-8. Nuclear factor kappa B (NF-kappaB) and nuclear factor IL-6 (NFIL-6) are primary mediators of IL-8 expression. We studied the transcriptional regulation of IL-8 in the ECV304 and 293T cell lines and found that the induction of IL-8 gene expression involved the activation of NF-kappaB (P = 0.001) and, to a lesser extent, the activation of NFIL-6 in ECV304 cells only. We next observed by the chromatin immunoprecipitation procedure in vivo acetylation of core histones bound to the IL-8 promoter after D2V infection. IL-8 produced by infected monocytes and also IL-8 that may be produced by endothelial or other epithelial cells is associated with the hyperacetylation of histones bound to the IL-8 promoter in addition to the activation of transcription by NF-kappaB. We hypothesize that the overall increase in IL-8 synthesis observed in this in vitro study may play a role in the pathogenesis of the plasma leakage seen in dengue hemorrhagic fever and dengue shock syndrome.