口蹄疫病毒3D蛋白促进TLR3介导的Ⅰ型干扰素产生

doi:10.11843/j.issn.0366-6964.2020.08.016

摘要/Abstract

摘要： 口蹄疫是由口蹄疫病毒（FMDV）引起的急性、热性、高度接触性传染病。口蹄疫病毒感染宿主引起一系列严重的炎症反应，而TLR3通路是介导细胞炎性反应的主要途径之一。为研究口蹄疫病毒蛋白对TLR3通路的影响，本研究首先用双荧光素酶报告系统筛选影响TLR3通路的FMDV蛋白；接着用Q-PCR试验验证筛出来的候选蛋白对TLR3通路下游基因表达水平的影响；并用免疫共沉淀试验验证与候选蛋白有相互作用的TLR3通路蛋白；最后用Western blot试验检测候选蛋白对TLR3通路下游分子磷酸化水平的影响。双荧光素酶报告系统结果显示，口蹄疫病毒3D蛋白促进TLR3通路介导的Ⅰ型干扰素的产生并呈剂量依赖性，Q-PCR试验表明，3D能够促进TLR3通路下游基因表达水平；免疫共沉淀试验表明，FMDV 3D与TLR3有相互作用；Western blot试验进一步显示，过表达3D能够促进TLR3下游分子的磷酸化水平。综上，口蹄疫病毒3D蛋白能促进TLR3介导的Ⅰ型干扰素的产生，从而调控天然免疫反应。

关键词: 口蹄疫病毒, 天然免疫, TLR3, 3D蛋白

Abstract: Foot-and-mouth disease (FMD) caused by FMD virus (FMDV) is an acute, hot and highly contagious disease. FMDV causes a series of severe inflammatory response, and TLR3 pathway is one of the major cellular inflammatory. In order to study the effect of FMDV protein on TLR3 pathway, we screened the FMDV proteins affecting on TLR3 pathway by reporter assays. Then candidate protein effect on transcription of TLR3 pathway downstream genes was verified by Q-PCR assay. TLR3 signaling pathway proteins interacted with candidate protein was further verified by co-immunocoprecipitation. Finally, we detected phosphorylation level of TLR3 pathway downstream proteins in candidate protein-overexpressed 293-TLR3 cells by Western blot. The luciferase assays and Q-PCR results showed that FMDV 3D potentiated poly (I:C)-triggered TLR3 signaling pathway and transcription of TLR3 pathway downstream genes, respectively. In addition, co-immunocoprecipitation showed that FMDV 3D interacted with TLR3. Furthermore, FMDV 3D increased the phosphorylation level of TLR3 pathway downstream protein. In conclusion, FMDV 3D potentiated type Ⅰ interferon mediated by TLR3 pathway, which regulate innate immune response.

Key words: FMDV, innate immune, TLR3, 3D protein

中图分类号:

S852.659.6

李露露, 张敬, 李丹, 郑海学. 口蹄疫病毒3D蛋白促进TLR3介导的Ⅰ型干扰素产生[J]. 畜牧兽医学报, 2020, 51(8): 1923-1931.

LI Lulu, ZHANG Jing, LI Dan, ZHENG Haixue. Foot-and-Mouth Disease Virus 3D Protein Positively Regulates the Production of InterferonⅠMediated by TLR3[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(8): 1923-1931.

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