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采用基于无标记的 DIA 方法定量分析 MARC-145 细胞感染墨西哥猪繁殖与呼吸综合征病毒株的蛋白质组。

Quantitative Proteomic Analysis of MARC-145 Cells Infected with a Mexican Porcine Reproductive and Respiratory Syndrome Virus Strain Using a Label-Free Based DIA approach.

机构信息

Unidad de Genómica, Proteómica y Metabolómica (UGPM), LaNSE, Cinvestav-IPN, Ciudad de México C.P. 07360, México.

Mass Spectrometry Applications & Development, SpectraMass Limited, Campinas, São Paulo 13088-130, Brazil.

出版信息

J Am Soc Mass Spectrom. 2020 Jun 3;31(6):1302-1312. doi: 10.1021/jasms.0c00134. Epub 2020 May 21.

DOI:10.1021/jasms.0c00134
PMID:32379441
Abstract

Porcine reproductive and respiratory syndrome (PRRS) is an infectious disease characterized by severe reproductive failure in sows, acute respiratory disorders in growing pigs, and high mortality in piglets. The causative agent of this syndrome is the PRRS virus (PRRSV), an RNA virus belonging to the Arteriviridae family. To date, several quantitative approaches of proteomics have been applied to analyze the gene expression profiles during PRRSV infection in PAMs and MARC-145 cells, and few proteins have been consistent among independent studies, probably due to the differences in the levels of virulence of different PRRSV strains used and/or due to analytical conditions. In this study, total proteins isolated from noninfected and infected MARC-145 cells with a Mexican PRRSV strain were relatively quantified using label-free based DIA approach in combination with ion-mobility separation. As a result, 1456 quantified proteins were found to be shared between the control and infected samples. Afterward, these proteins were filtered, and 699 of them were considered without change. Also, 17 proteins were up-regulated and 19 proteins were down-regulated during the PRSSV infection. Bioinformatic analysis revealed that many of the differentially expressed proteins are involved in processes like antigen processing, presentation of antigens, response to viruses, response to IFNs, and innate immune response, among others. The present work is the first one which provides a detailed proteomic analysis through label-free based DIA approach in MARC-145 cells during the infection with a Mexican PRRSV strain.

摘要

猪繁殖与呼吸综合征(PRRS)是一种传染病,其特征是母猪严重繁殖失败、生长猪急性呼吸道疾病和仔猪高死亡率。该综合征的病原体是 PRRS 病毒(PRRSV),一种属于动脉炎病毒科的 RNA 病毒。迄今为止,已经应用了几种定量蛋白质组学方法来分析 PAMs 和 MARC-145 细胞中 PRRSV 感染时的基因表达谱,并且很少有蛋白质在独立研究中是一致的,这可能是由于使用的不同 PRRSV 毒株的毒力水平不同,或者由于分析条件不同。在这项研究中,使用无标记的 DIA 方法结合离子迁移分离,对未感染和感染墨西哥 PRRSV 株的 MARC-145 细胞中的总蛋白进行相对定量。结果,在对照和感染样本之间发现了 1456 个可定量的蛋白质。之后,对这些蛋白质进行过滤,其中 699 个被认为没有变化。此外,在 PRSSV 感染期间,有 17 种蛋白质上调,19 种蛋白质下调。生物信息学分析表明,许多差异表达的蛋白质参与抗原加工、抗原呈递、对病毒的反应、对 IFNs 的反应和先天免疫反应等过程。本工作是首次通过无标记的 DIA 方法在 MARC-145 细胞中对感染墨西哥 PRRSV 株的细胞进行详细的蛋白质组学分析。

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