Virulent and Vaccine Strains of Streptococcus equi ssp. zooepidemicus Have Different Influences on Phagocytosis and Cytokine Secretion of Macrophages.
Virulent and Vaccine Strains of Streptococcus equi ssp. zooepidemicus Have Different Influences on Phagocytosis and Cytokine Secretion of Macrophages.
复制标题
DOI：
10.1021/acs.jproteome.6b00571
复制
发表时间：
2017-01
影响因子：
4.4
通讯作者：
Peng Jie;Ma Zhe;Hu Chengwei;Lin Huixing;Z. Hui;Luo Chengping;Fan Hong-jie
中科院分区：
文献类型：
--
作者：
Peng Jie;Ma Zhe;Hu Chengwei;Lin Huixing;Z. Hui;Luo Chengping;Fan Hong-jie
猪链球菌病是我国养猪业面临的重大威胁，马链球菌（Streptococcus equi ssp.）动物寄生虫（Zooprophylmicus，SEZ）是主要的病原体之一。SEZ ATCC 35246是经典强毒株，而SEZ ST 171是中国减毒疫苗株。在这项研究中，我们采用稳定同位素标记的氨基酸在细胞培养和液相色谱-质谱（LC-MS），以确定这两种菌株感染的巨噬细胞的差异反应。87个上调蛋白和135个下调蛋白被鉴定。蛋白质组学的结果进行了验证，通过实时聚合酶链反应的10个选定的基因和蛋白质印迹法的三个蛋白质。分析所有差异丰度蛋白质的基因本体论和京都基因和基因组百科全书注释。某些下调的蛋白质与免疫功能有关，而上调的蛋白质则与细胞膜和细胞骨架的调节有关。检测SEZ ATCC 35246和ST 171感染Raw 264. 7细胞后的吞噬率和细胞因子基因转录水平，验证生物信息学结果。这些结果表明，对巨噬细胞吞噬功能和细胞因子表达的不同影响可能解释了SEZ ATCC 35246和ST 171感染的不同表型。本研究为阐明SEZ ST 171和SEZ ATCC 35246的免疫应答机制提供了线索，为进一步研究SEZ ST 171和SEZ ATCC 35246的治疗和疫苗开发提供了新的靶点。
Swine streptococcosis is a significant threat to the Chinese pig industry, and Streptococcus equi ssp. zooepidemicus (SEZ) is one of the major pathogens. SEZ ATCC35246 is a classical virulent strain, while SEZ ST171 is a Chinese attenuated vaccine strain. In this study, we employed stable isotope labeling by amino acids in cell culture and liquid chromatography-mass spectrometry (LC-MS) to determine the differential response of macrophages to infection by these two strains. Eighty-seven upregulated proteins and 135 downregulated proteins were identified. The proteomic results were verified by real-time polymerase chain reaction for 10 chosen genes and Western blotting for three proteins. All differentially abundant proteins were analyzed for their Gene Ontology and Kyoto Encyclopedia of Genes and Genomes annotations. Certain downregulated proteins were associated with immunity functions, and the upregulated proteins were related to cytomembrane and cytoskeleton regulation. The phagocytosis rate and cytokine genes transcription in Raw264.7 cells during SEZ ATCC35246 and ST171 infection were detected to confirm the bioinformatics results. These results showed that different effects on macrophage phagocytosis and cytokine expression might explain the different phenotypes of SEZ ATCC35246 and ST171 infection. This research provided clues to the mechanisms of host immunity responses to SEZ ST171and SEZ ATCC35246, which could identify potential therapy and vaccine development targets.