Host gene expression profiling of dengue virus infection in cell lines and patients.
Host gene expression profiling of dengue virus infection in cell lines and patients.
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DOI：
10.1371/journal.pntd.0000086
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发表时间：
2007-11-21
影响因子：
3.8
通讯作者：
Hibberd ML
中科院分区：
文献类型：
--
作者：
Fink J;Gu F;Ling L;Tolfvenstam T;Olfat F;Chin KC;Aw P;George J;Kuznetsov VA;Schreiber M;Vasudevan SG;Hibberd ML
尽管登革热相关疾病的严重性，估计每年有5000 - 1亿例登革热病例和250，000 - 500，000例登革出血热/登革休克综合征，但对登革热发病机制的明确理解仍然是难以捉摸的。由于缺乏动物疾病模型和登革热感染中复杂的免疫相互作用，宿主反应和免疫发病机制的研究是困难的。基因组学技术、微阵列技术和高通量定量PCR技术的发展使研究人员能够在更广泛的范围内研究基因表达的变化。因此，我们使用这种方法来研究登革热病毒感染的细胞系和登革热患者的宿主反应。利用微阵列和高通量定量PCR方法监测细胞系感染模型和登革热患者血液样本中宿主对登革热病毒复制的反应，我们确定了三条主要途径沿着差异表达基因：NF-κB启动的免疫反应，I型干扰素（IFN）和泛素蛋白酶体途径。其中上调最高的基因是趋化因子IP-10和I-TAC，两者都是CXCR 3受体的配体。通过ELISA证实了10名患者在发热早期外周血中IP-10和I-TAC的表达增加。IFN途径中的一个高度上调的基因viperin在A549细胞中过表达，导致病毒复制显著减少。泛素-蛋白酶体途径中基因的上调促使蛋白酶体抑制剂MG-132和ALLN的测试，这两种抑制剂都能减少病毒复制。无偏见的基因表达分析已经确定了新的宿主基因与登革热感染，我们已经验证了功能研究。我们发现，宿主反应的某些部分可用作该疾病的潜在生物标志物，而其他部分可用于控制登革热病毒复制，从而代表药物治疗的可行靶点。登革热是影响人类的最普遍的蚊媒病毒性疾病，但目前还没有针对该疾病的药物治疗方法，也没有任何经过验证的宿主靶点进行治疗干预。利用微阵列技术监测几乎每个人类基因的反应，我们的目标是确定人类在感染过程中与登革热病毒相互作用的方式，以发现可用于控制病毒复制的新治疗靶点。从激活的基因中，我们确定了体外和体内感染共有的三条途径：NF-κB启动的免疫途径、I型干扰素途径和泛素蛋白酶体途径。我们接下来发现，抑制泛素蛋白酶体途径，或激活I型干扰素途径，导致病毒复制的显着抑制。然而，抑制NF-κB启动的免疫途径对病毒复制没有影响。我们认为，靶向泛素蛋白酶体途径的药物可能被证明在杀死登革病毒方面是有效的，并且如果用于治疗，可以改善登革疾病的临床结果。
Despite the seriousness of dengue-related disease, with an estimated 50–100 million cases of dengue fever and 250,000–500,000 cases of dengue hemorrhagic fever/dengue shock syndrome each year, a clear understanding of dengue pathogenesis remains elusive. Because of the lack of a disease model in animals and the complex immune interaction in dengue infection, the study of host response and immunopathogenesis is difficult. The development of genomics technology, microarray and high throughput quantitative PCR have allowed researchers to study gene expression changes on a much broader scale. We therefore used this approach to investigate the host response in dengue virus-infected cell lines and in patients developing dengue fever. Using microarray and high throughput quantitative PCR method to monitor the host response to dengue viral replication in cell line infection models and in dengue patient blood samples, we identified differentially expressed genes along three major pathways; NF-κB initiated immune responses, type I interferon (IFN) and the ubiquitin proteasome pathway. Among the most highly upregulated genes were the chemokines IP-10 and I-TAC, both ligands of the CXCR3 receptor. Increased expression of IP-10 and I-TAC in the peripheral blood of ten patients at the early onset of fever was confirmed by ELISA. A highly upregulated gene in the IFN pathway, viperin, was overexpressed in A549 cells resulting in a significant reduction in viral replication. The upregulation of genes in the ubiquitin-proteasome pathway prompted the testing of proteasome inhibitors MG-132 and ALLN, both of which reduced viral replication. Unbiased gene expression analysis has identified new host genes associated with dengue infection, which we have validated in functional studies. We showed that some parts of the host response can be used as potential biomarkers for the disease while others can be used to control dengue viral replication, thus representing viable targets for drug therapy. Dengue is the most prevalent mosquito-born viral disease affecting humans, yet there is, at present, no drug treatment for the disease nor are there any validated host targets for therapeutic intervention. Using microarray technology to monitor the response of virtually every human gene, we aimed to identify the ways in which humans interact with dengue virus during infection in order to discover new therapeutic targets that could be exploited to control viral replication. From the activated genes, we identified three pathways common to in vitro and in vivo infection; the NF-κB initiated immune pathway, the type I interferon pathway, and the ubiquitin proteasome pathway. We next found that inhibiting the ubiquitin proteasome pathway, or activating the type I interferon pathway, resulted in significant inhibition of viral replication. However, inhibiting the NF-κB initiated immune pathway had no effect on viral replication. We suggest that drugs that target the ubiquitin proteasome pathway may prove effective at killing the dengue virus, and, if used therapeutically, improve clinical outcome in dengue disease.
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DOI：
10.1084/jem.184.3.963
发表时间：
1996-09-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Loetscher M;Gerber B;Loetscher P;Jones SA;Piali L;Clark-Lewis I;Baggiolini M;Moser B
通讯作者：
Moser B
影响因子：
3.8
作者：
Matsuda, T;Almasan, A;Mori, N
通讯作者：
Mori, N
DOI：
10.1084/jem.187.12.2009
发表时间：
1998-06-15
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Cole KE;Strick CA;Paradis TJ;Ogborne KT;Loetscher M;Gladue RP;Lin W;Boyd JG;Moser B;Wood DE;Sahagan BG;Neote K
通讯作者：
Neote K
影响因子：
5.4
作者：
Gao, L;Tu, H;Lai, MMC
通讯作者：
Lai, MMC
影响因子：
56.9
作者：
AUPHAN, N;DIDONATO, JA;KARIN, M
通讯作者：
KARIN, M