Integrated molecular signature of disease: Analysis of influenza virus-infected macaques through functional genomics and proteomics
Integrated molecular signature of disease: Analysis of influenza virus-infected macaques through functional genomics and proteomics
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DOI：
10.1128/jvi.00851-06
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发表时间：
2006-11-01
影响因子：
5.4
通讯作者：
Katze, M. G.
中科院分区：
文献类型：
--
作者：
Baas, T.;Baskin, C. R.;Katze, M. G.
最近在人类中爆发的禽流感强调了改进非人类灵长类流感发病机制模型的必要性。为了进一步建立猕猴模型，我们对流感病毒感染的猕猴进行了体内基因组学实验，重点研究了感染流感病毒后第2天的先天免疫反应，以及存在病毒遗传物质的受影响肺组织中的基因表达。最后，我们试图确定全血中早期感染的特征基因。为此，我们用重组A/Texas/36/91流感病毒感染了6只辫尾猕猴(Macaca Nomestrina)，并用假疫苗接种了3只对照动物。分别于感染后第2、4、7天处死1只对照动物和2只实验动物。肺组织用于病理学、基因表达谱和蛋白质组学。每隔一天从每只动物身上采集血液进行基因组学检测，直到实验终点。大体和显微病理学、免疫组织化学、阵列病毒基因表达和/或实时定量逆转录-聚合酶链式反应证实了所有实验动物的成功但轻微的感染。基因组实验使用猕猴特异的寡核苷酸阵列进行，高通量蛋白质组学在mRNA和蛋白质水平上揭示了宿主对感染的反应。我们的数据显示，在流感病毒诱导的损伤中，基于病毒mRNA的存在或不存在，区域内的基因表达存在显著差异。在接种后第2天，我们还在外周血白细胞和肺组织中发现了紧密共调控的基因。后一项发现开启了使用全血基因表达阵列检测感染的可能性，暴露后但在出现症状或脱落之前。
Recent outbreaks of avian influenza in humans have stressed the need for an improved nonhuman primate model of influenza pathogenesis. In order to further develop a macaque model, we expanded our previous in vivo genomics experiments with influenza virus-infected macaques by focusing on the innate immune response at day 2 postinoculation and on gene expression in affected lung tissue with viral genetic material present. Finally, we sought to identify signature genes for early infection in whole blood. For these purposes, we infected six pigtailed macaques (Macaca nemestrina) with reconstructed influenza A/Texas/36/91 virus and three control animals with a sham inoculate. We sacrificed one control and two experimental animals at days 2, 4, and 7 postinfection. Lung tissue was harvested for pathology, gene expression profiling, and proteomics. Blood was collected for genomics every other day from each animal until the experimental endpoint. Gross and microscopic pathology, immunohistochemistry, viral gene expression by arrays, and/or quantitative real-time reverse transcription-PCR confirmed successful yet mild infections in all experimental animals. Genomic experiments were performed using macaque-specific oligonucleotide arrays, and high-throughput proteomics revealed the host response to infection at the mRNA and protein levels. Our data showed dramatic differences in gene expression within regions in influenza virus-induced lesions based on the presence or absence of viral mRNA. We also identified genes tightly coregulated in peripheral white blood cells and in lung tissue at day 2 postinoculation. This latter finding opens the possibility of using gene expression arrays on whole blood to detect infection after exposure but prior to onset of symptoms or shedding.