Central Laboratory, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, China.
Department of General Surgery, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, China.
Methods Mol Biol. 2025;2854:93-106. doi: 10.1007/978-1-0716-4108-8_11.
As an interferon-stimulating factor protein, STING plays a role in the response and downstream liaison in antiviral natural immunity. Upon viral invasion, the immediate response of STING protein leads to a series of changes in downstream proteins, which ultimately leads to an antiviral immune response in the form of proinflammatory cytokines and type I interferons, thus triggering an innate immune response, an adaptive immune response in vivo, and long-term protection of the host. In the field of antiviral natural immunity, it is particularly important to rigorously and sequentially probe the dynamic changes in the antiviral natural immunity connector protein STING caused by the entire anti-inflammatory and anti-pathway mechanism and the differences in upstream and downstream proteins. Traditionally, proteomics technology has been validated by detecting proteins in a 2D platform, for which it is difficult to sensitively identify changes in the nature and abundance of target proteins. With the development of mass spectrometry (MS) technology, MS-based proteomics has made important contributions to characterizing the dynamic changes in the natural immune proteome induced by viral infections. MS analytical techniques have several advantages, such as high throughput, rapidity, sensitivity, accuracy, and automation. The most common techniques for detecting complex proteomes are liquid chromatography (LC) and mass spectrometry (MS). LC-MS (Liquid Chromatography-Mass Spectrometry), which combines the physical separation capability of LC and the mass analysis capability of MS, is a powerful technique mainly used for analyzing the proteome of cells, tissues, and body fluids. To explore the combination of traditional proteomics techniques such as Western blotting, Co-IP (co-Immunoprecipitation), and the latest LC-MS methods to probe the anti-inflammatory pathway and the differential changes in upstream and downstream proteins induced by the antiviral natural immune junction protein STING.
作为干扰素刺激因子蛋白，STING 在抗病毒天然免疫的反应和下游联络中发挥作用。病毒入侵时，STING 蛋白的即刻反应导致下游蛋白发生一系列变化，最终以促炎细胞因子和 I 型干扰素的形式引发抗病毒免疫反应，从而触发先天免疫反应、体内适应性免疫反应和宿主的长期保护。在抗病毒天然免疫领域，严格、有序地探测抗病毒天然免疫接头蛋白 STING 被整个抗炎和抗途径机制以及上下游蛋白差异引起的动态变化非常重要。传统上，蛋白质组学技术通过在 2D 平台上检测蛋白质得到验证，因此很难敏感地识别目标蛋白质的性质和丰度变化。随着质谱（MS）技术的发展，基于 MS 的蛋白质组学为描述病毒感染诱导的天然免疫蛋白质组的动态变化做出了重要贡献。MS 分析技术具有高通量、快速、灵敏、准确和自动化等优点。检测复杂蛋白质组的最常见技术是液相色谱（LC）和质谱（MS）。LC-MS（液相色谱-质谱联用）将 LC 的物理分离能力和 MS 的质量分析能力相结合，是一种强大的技术，主要用于分析细胞、组织和体液的蛋白质组。为了探索传统蛋白质组学技术（如 Western blot、Co-IP（共免疫沉淀））与最新的 LC-MS 方法的结合，以探测抗炎途径和抗病毒天然免疫接头蛋白 STING 诱导的上下游蛋白的差异变化。
