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猪肠系膜淋巴结感染鼠伤寒沙门氏菌后的蛋白质组学分析。

Proteomic analysis of porcine mesenteric lymph-nodes after Salmonella typhimurium infection.

机构信息

Grupo de Genómica y Mejora Animal, Departamento de Genética, Universidad de Córdoba, Campus de Rabanales, 14071 Córdoba, Spain.

出版信息

J Proteomics. 2012 Jul 19;75(14):4457-70. doi: 10.1016/j.jprot.2012.03.045. Epub 2012 Apr 3.

Abstract

In this study we employed for the first time an in vivo approach coupled to DIGE-based proteomics to explore the response of porcine mesenteric lymph nodes (MLN) to Salmonella typhimurium infection. MLN samples were collected from four control and twelve infected pigs (at 1, 2 and 6 days post infection) for histological analysis, protein and RNA purification. Afterwards, expressed proteins were screened by differential in gel analysis and data were analyzed by bioinformatic tools to generate interaction networks, and identify enriched signaling pathways and biological annotations. S. typhimurium labeling in tissue and phagocyte infiltration were analyzed by immunohistochemistry and RNA was employed to determine the relative expression of immune-related genes by quantitative RNA analysis. The proteome response of porcine MLN to infection was associated to the induction of processes such as phagocyte infiltration, cytoskeleton remodeling and pyroptosis. Moreover, our results suggest that S. typhimurium antigens are cross-presented via MHC-I in a proteasome-dependent manner in porcine MLN. Since pathogen burden in tissue was noticeably reduced at the end of the time course, we infer that host innate and adaptive immunity act in association in MLN to control S. typhimurium dissemination in swine infections.

摘要

在这项研究中,我们首次采用体内方法结合 DIGE 蛋白质组学来研究猪肠系膜淋巴结 (MLN) 对鼠伤寒沙门氏菌感染的反应。从 4 只对照猪和 12 只感染猪(感染后 1、2 和 6 天)采集 MLN 样本,进行组织学分析、蛋白质和 RNA 纯化。然后,通过差异凝胶分析筛选表达蛋白,并通过生物信息学工具分析数据,生成互作网络,鉴定富集的信号通路和生物学注释。通过免疫组织化学分析组织中的鼠伤寒沙门氏菌标记和吞噬细胞浸润,并用定量 RNA 分析确定免疫相关基因的相对表达。猪 MLN 对感染的蛋白质组反应与吞噬细胞浸润、细胞骨架重塑和细胞焦亡等过程的诱导有关。此外,我们的结果表明,鼠伤寒沙门氏菌抗原通过 MHC-I 以依赖蛋白酶体的方式在猪 MLN 中交叉呈递。由于组织中的病原体负荷在时间过程结束时明显降低,我们推断宿主固有和适应性免疫在 MLN 中协同作用,以控制猪感染中鼠伤寒沙门氏菌的传播。

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