A two-phase innate host response to alphavirus infection identified by mRNP-tagging in vivo
A two-phase innate host response to alphavirus infection identified by mRNP-tagging in vivo
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DOI：
10.1371/journal.ppat.0030199
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发表时间：
2007-12-01
期刊：
影响因子：
6.7
通讯作者：
Johnston, Robert E.
中科院分区：
文献类型：
--
作者：
Konopka, Jennifer L.;Penalva, Luiz O.;Johnston, Robert E.
病毒发病机理的基本概念是感染诱导宿主细胞内、特定组织内或整个动物内的特定变化。这些变化反映在感染过程中明显改变的转录模式的级联中。然而，这种级联反应在体内的阐明一直受到限制，一般无法区分的变化发生在少数受感染的细胞从周围未感染的细胞。为了规避传统基因表达谱分析方法的固有限制，实施了一种创新的mRNP标记技术，以在体外以及体内委内瑞拉马脑炎病毒（VEE）感染后从感染细胞中特异性分离宿主mRNA。这种技术有助于直接表征宿主防御反应，特别是在第一个细胞感染VEE，而同时总RNA分析评估感染和未感染细胞的集体反应。结果是在初级树突状细胞以及引流淋巴结（小鼠模型中最初的靶组织）中对VEE感染的早期反应的独特，多方面的特征。揭示了复杂相互作用的动态环境，并提出了两步先天反应，其中感染细胞中宿主基因的子集的激活随后导致周围未感染细胞的激活。我们的研究结果表明，应用病毒mRNP标记系统，如这里介绍的，将有助于更详细地了解高度协调的宿主对感染因子的反应。
A concept fundamental to viral pathogenesis is that infection induces specific changes within the host cell, within specific tissues, or within the entire animal. These changes are reflected in a cascade of altered transcription patterns evident during infection. However, elucidation of this cascade in vivo has been limited by a general inability to distinguish changes occurring in the minority of infected cells from those in surrounding uninfected cells. To circumvent this inherent limitation of traditional gene expression profiling methods, an innovative mRNP-tagging technique was implemented to isolate host mRNA specifically from infected cells in vitro as well as in vivo following Venezuelan equine encephalitis virus (VEE) infection. This technique facilitated a direct characterization of the host defense response specifically within the first cells infected with VEE, while simultaneous total RNA analysis assessed the collective response of both the infected and uninfected cells. The result was a unique, multifaceted profile of the early response to VEE infection in primary dendritic cells, as well as in the draining lymph node, the initially targeted tissue in the mouse model. A dynamic environment of complex interactions was revealed, and suggested a two-step innate response in which activation of a subset of host genes in infected cells subsequently leads to activation of the surrounding uninfected cells. Our findings suggest that the application of viral mRNP-tagging systems, as introduced here, will facilitate a much more detailed understanding of the highly coordinated host response to infectious agents.