猪繁殖与呼吸综合征病毒感染猪肺泡巨噬细胞天然免疫信号通路相关分子变化分析

doi:10.11843/j.issn.0366-6964.2017.07.013

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (7): 1288-1299.doi: 10.11843/j.issn.0366-6964.2017.07.013

猪繁殖与呼吸综合征病毒感染猪肺泡巨噬细胞天然免疫信号通路相关分子变化分析

李华玮^1,4, 陈鑫鑫², 赵孟孟³, 周恩民¹, 乔松林^2*, 张改平^1,2,3*

1. 西北农林科技大学动物医学院, 杨凌 712100;
2. 河南省农业科学院动物免疫学重点实验室, 郑州 450002;
3. 河南农业大学牧医工程学院, 郑州 450002;
4. 河南牧业经济学院生物工程学院, 郑州 450046

收稿日期:2017-01-09 出版日期:2017-07-23 发布日期:2017-07-23
通讯作者: 张改平(1960-),院士,博士生导师,主要从事动物免疫学及动物病毒分子致病机制研究,E-mail:zhanggaiping2003@163.com;乔松林(1970-),河南鲁山人,研究员,主要从事预防兽医学研究,E-mail:cdj565@gmail.com
作者简介:李华玮(1981-),女,河南郑州人,博士生,主要从事动物病毒分子生物学与分子免疫学研究,E-mail:centrosome@126.com
基金资助:
国家自然科学基金重大项目（31490601）；国家自然科学基金（31502043）；国家生猪产业技术体系岗位科学家项目（CARS-36）；河南省生猪产业技术体系创新团队项目（S2012-06）

Analysis of Related Molecular Changes of Innate Immune Signaling Pathway in Porcine Reproductive and Respiratory Syndrome Virus Infected PAMs

LI Hua-wei^1,4, CHEN Xin-xin², ZHAO Meng-meng³, ZHOU En-min¹, QIAO Song-lin^2*, ZHANG Gai-ping^1,2,3*

1. College of Veterinary Medicine, Northwest A & F University, Yangling 712100, China;
2. The Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China;
3. College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China;
4. The Bioengineering Department, Henan University of Animal Husbandry and Economy, Zhengzhou 450046, China

Received:2017-01-09 Online:2017-07-23 Published:2017-07-23

摘要/Abstract

摘要：

利用蛋白质组学的方法对猪繁殖与呼吸综合征病毒（porcine reproductive and respiratory syndrome virus，PRRSV）感染猪肺泡巨噬细胞（pulmonary macrophage，PAM）后的蛋白质动态变化进行探讨，为阐明病毒感染后引起的复杂免疫学反应及为该病的防控提供理论依据。通过Label-free LC-MS/MS方法定量鉴定PRRSV感染PAMs后差异表达细胞蛋白质。结果显示，共鉴定出430个显著差异表达蛋白质，其中包括171个上调表达蛋白质和259个下调表达蛋白质，这些蛋白质与细胞抗病毒天然免疫及抗原递呈等相关。值得关注的是，许多上调表达蛋白质都富集在与天然免疫相关的NF-κB、MAPK及RIG-Ⅰ等信号通路上，包括STAT1/2、OAS1/2、Mx1/2、ISG15/20、IFIT1/2/3/5等；下调蛋白质主要富集在MHCⅠ和MHCⅡ分子相关信号通路上。荧光定量PCR和Western blot结果证实，PRRSV感染导致了STAT1/2、OAS1/2、Mx1、CD14、ISG和IFIT等的转录或表达，这为进一步阐述PRRSV的感染机制奠定基础。

Abstract:

This research was conducted to analyze the proteomic dynamic proteins of PRRSV-infected PAMs. In this study, Label-free LC-MS/MS was employed to quantitatively identify the differentially expressed proteins between PRRSV-infected PAMs and uninfected PAMs. Results were as follows:430 significantly altered proteins were identified after PRRSV infection. Among them, 171 proteins were up-regulated, and 259 proteins were down-regulated. These differentially expressed proteins are related to innate immunity, virus response and antigen presentation etc. Interestingly, many upregulated proteins were enriched in NF-κB, MAPK and RIG-1 signal pathway, including STAT1/2, OAS1/2, Mx1/2, ISG15/20, IFIT1/2/3/5; some proteins were enriched in MHCⅠand MHCⅡrelated signal pathways. Real-time quantitative reverse transcription polymerase chain reaction and Western blot analysis verifiedSTAT 1/2, OAS 1/2, CD 14, ISG, Mx1, IFIT genes and Mx1, IFIT3 proteins were significant up-regulated after PRRSV infection which were consistent with LC-MS/MS results. This study may accelerate our understanding of the mechanisms of PRRSV infection.

中图分类号:

S852.659.6

李华玮, 陈鑫鑫, 赵孟孟, 周恩民, 乔松林, 张改平. 猪繁殖与呼吸综合征病毒感染猪肺泡巨噬细胞天然免疫信号通路相关分子变化分析[J]. 畜牧兽医学报, 2017, 48(7): 1288-1299.

LI Hua-wei, CHEN Xin-xin, ZHAO Meng-meng, ZHOU En-min, QIAO Song-lin, ZHANG Gai-ping. Analysis of Related Molecular Changes of Innate Immune Signaling Pathway in Porcine Reproductive and Respiratory Syndrome Virus Infected PAMs[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(7): 1288-1299.

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猪繁殖与呼吸综合征病毒感染猪肺泡巨噬细胞天然免疫信号通路相关分子变化分析

Analysis of Related Molecular Changes of Innate Immune Signaling Pathway in Porcine Reproductive and Respiratory Syndrome Virus Infected PAMs

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