Rus Florentina, Morlock Kayla, Silverman Neal, Pham Ngoc, Kotwal Girish J, Marshall William L
Division of Infectious Disease, Department of Medicine, University of Massachusetts School of Medicine, Worcester, MA, USA.
Methods Mol Biol. 2012;890:273-88. doi: 10.1007/978-1-61779-876-4_16.
Innate immune recognition of pathogens is critical to the prompt control of infections, permitting the host to survive to develop long-term immunity via an adaptive immune response. Poxviruses encode a family of proteins that inhibit signaling by Toll-like receptors to their downstream signaling components, severely limiting nuclear translocation of transcription factors such as IRF3 and NF-κB and thereby decreasing production of host interferons and cytokines. We describe bioinformatics techniques for identifying candidate poxviral inhibitors of the innate immune response based on similarity to the family of proteins that includes A52, A46, and N1. Robust luciferase assays can determine whether a given poxviral gene affects innate immune signaling, and in combination with other approaches can identify the cellular targets of poxviral innate immune evasion genes. Because apoptosis is an innate immune response of the cell to viral infection, assays for identifying poxviral genes that inhibit apoptosis can also be employed. Novel poxviral innate immune inhibitors are being identified via several approaches and these techniques promise to identify further complexities in the way that poxviruses interact with the host innate immune system.
对病原体的天然免疫识别对于迅速控制感染至关重要，它使宿主能够存活下来，通过适应性免疫反应产生长期免疫力。痘病毒编码一类蛋白质，这类蛋白质可抑制Toll样受体向其下游信号成分的信号传导，严重限制转录因子（如IRF3和NF-κB）的核转位，从而减少宿主干扰素和细胞因子的产生。我们描述了基于与包括A52、A46和N1在内的蛋白质家族的相似性来鉴定天然免疫反应的候选痘病毒抑制剂的生物信息学技术。可靠的荧光素酶测定法可以确定给定的痘病毒基因是否影响天然免疫信号传导，并且与其他方法相结合可以鉴定痘病毒天然免疫逃避基因的细胞靶点。由于细胞凋亡是细胞对病毒感染的天然免疫反应，因此也可以采用鉴定抑制细胞凋亡的痘病毒基因的测定法。通过几种方法正在鉴定新型痘病毒天然免疫抑制剂，这些技术有望揭示痘病毒与宿主天然免疫系统相互作用方式的更多复杂性。
