Falcinelli Shane, Gowen Brian B, Trost Brett, Napper Scott, Kusalik Anthony, Johnson Reed F, Safronetz David, Prescott Joseph, Wahl-Jensen Victoria, Jahrling Peter B, Kindrachuk Jason
From the ‡Emerging Viral Pathogens Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA;
§Department of Animal, Dairy, and Veterinary Sciences, Utah State University, Logan, Utah, USA;
Mol Cell Proteomics. 2015 Mar;14(3):646-57. doi: 10.1074/mcp.M114.045443. Epub 2015 Jan 8.
The Syrian golden hamster has been increasingly used to study viral hemorrhagic fever (VHF) pathogenesis and countermeasure efficacy. As VHFs are a global health concern, well-characterized animal models are essential for both the development of therapeutics and vaccines as well as for increasing our understanding of the molecular events that underlie viral pathogenesis. However, the paucity of reagents or platforms that are available for studying hamsters at a molecular level limits the ability to extract biological information from this important animal model. As such, there is a need to develop platforms/technologies for characterizing host responses of hamsters at a molecular level. To this end, we developed hamster-specific kinome peptide arrays to characterize the molecular host response of the Syrian golden hamster. After validating the functionality of the arrays using immune agonists of defined signaling mechanisms (lipopolysaccharide (LPS) and tumor necrosis factor (TNF)-α), we characterized the host response in a hamster model of VHF based on Pichinde virus (PICV(1)) infection by performing temporal kinome analysis of lung tissue. Our analysis revealed key roles for vascular endothelial growth factor (VEGF), interleukin (IL) responses, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling, and Toll-like receptor (TLR) signaling in the response to PICV infection. These findings were validated through phosphorylation-specific Western blot analysis. Overall, we have demonstrated that hamster-specific kinome arrays are a robust tool for characterizing the species-specific molecular host response in a VHF model. Further, our results provide key insights into the hamster host response to PICV infection and will inform future studies with high-consequence VHF pathogens.
叙利亚金黄地鼠越来越多地被用于研究病毒性出血热(VHF)的发病机制和对策效果。由于VHF是一个全球健康问题,特征明确的动物模型对于治疗方法和疫苗的开发以及增进我们对病毒发病机制背后分子事件的理解都至关重要。然而,可用于在分子水平研究地鼠的试剂或平台匮乏,限制了从这个重要动物模型中提取生物学信息的能力。因此,需要开发在分子水平表征地鼠宿主反应的平台/技术。为此,我们开发了地鼠特异性激酶组肽阵列,以表征叙利亚金黄地鼠的分子宿主反应。在用具有明确信号传导机制的免疫激动剂(脂多糖(LPS)和肿瘤坏死因子(TNF)-α)验证阵列的功能后,我们通过对肺组织进行时间激酶组分析,在基于皮钦德病毒(PICV(1))感染的VHF地鼠模型中表征宿主反应。我们的分析揭示了血管内皮生长因子(VEGF)、白细胞介素(IL)反应、活化B细胞核因子κB(NF-κB)信号传导和Toll样受体(TLR)信号传导在对PICV感染的反应中的关键作用。这些发现通过磷酸化特异性蛋白质印迹分析得到验证。总体而言,我们证明了地鼠特异性激酶组阵列是在VHF模型中表征物种特异性分子宿主反应的强大工具。此外,我们的结果为地鼠宿主对PICV感染的反应提供了关键见解,并将为未来针对高致病性VHF病原体的研究提供信息。