摘要
为揭示肥大细胞抗口蹄疫病毒VP1-VP4蛋白的天然免疫作用,以重组口蹄疫病毒VP1-VP4蛋白刺激小鼠腹腔肥大细胞(Peritoneal mast cells,PMCs),用高通量ELISA芯片检测PMCs的蛋白质表达谱。结果显示,VP1-VP4蛋白刺激的PMCs(VP1-VP4组)表达CCL19、L-selectin、CCL17和TNF-α的水平极显著低于对照组(PMCs)(P<0.001),而VP1-VP4蛋白刺激经甘露糖受体(Mannose receptor,MR)抑制剂预处理的PMCs(MR组)表达CCL19、IL-15、IL-9、G-CSF和Galectin-1的水平则极显著高于对照组(P<0.01),IL-10表达水平也有显著升高(P<0.05)。MR组与VP1-VP4组相比,PMCs表达IL-10、IL-17、CCL20、IL-15、IL-9、L-selectin、CCL17、TNF-α和CCL19的水平极显著升高(P<0.01),CCL21和G-CSF的表达也显著高于VP1-VP4组(P<0.05)。生物信息学差异表达分析结果显示,与对照组相比,VP1-VP4组PMCs表达的L-selectin和CCL17为下调性差异表达蛋白(Log2(ratio)≤–1)。MR组与VP1-VP4相比,PMCs表达的CCL20、CCL19、L-selectin和IL-15为上调性差异表达蛋白(Log_2(ratio)≥1)。这表明,PMCs可自发分泌CCL19、L-selectin、CCL17和TNF-α,而VP1-VP4则对PMCs的天然免疫功能具有抑制作用。由于阻断MR后PMCs的蛋白质表达水平显著升高,所以VP1-VP4对小鼠PMCs的免疫抑制作用可能是由MR介导的。
To reveal the innate immunity of mast cells against recombinant VP1-VP4 protein of foot-and-mouth disease virus（FMDV）, mouse peritoneal mast cells（PMCs） were pulsed with recombinant VP1-VP4 protein. The supernatants harvested from PMCs cultures were applied to the high throughput ELISA array. Our results show that the expression levels of CCL19, L-selectin, CCL17, and TNF alpha released from PMCs pulsed with recombinant VP1-VP4 were significantly down-regulated compared with PMCs alone（P0.001）. Surprisingly, in comparison with PMCs alone, the expression levels of CCL19, IL-15, IL-9, G-CSF, and Galectin-1 in PMCs with the mannose receptor（MR） inhibitor were significantly up-regulated（P0.01）, and the expression level of IL-10 was also remarkably up-regulated（P0.05）. Importantly, the protein expression levels in PMCs treated with MR inhibitor were higher than PMCs pulsed with VP1-VP4, including IL-10, IL-17, CCL20, IL-15, IL-9, L-selectin, CCL17, TNF alpha, and CCL19（P0.01） as well as CCL21, and G-CSF（P0.05）. Differential expression analysis in bioinformatics shows that both L-selectin and CCL17 were recognized as differentially expressed protein molecules（Log2（ratio）≤–1） when compared with PMCs alone. Furthermore, the up-regulation of the expression levels of CCL20, CCL19, L-selectin, and IL-15 in PMCs treated with MR inhibitor was defined as differential expression（Log_2（ratio）≥1）. These data indicate that PMCs are capable of secreting CCL19, L-selectin, CCL17, and TNF alpha spontaneously and the recombinant VP1-VP4 has an inhibitive potential to PMCs during their performance of innate immune response. Given the protein expression levels from PMCs pre-treated with MR inhibitor were significantly increased, it can be deduced that immunosuppression of FMDV is presumably initiated by the VP1 recognition of MR on mast cells.
出处
《生物工程学报》
CAS
CSCD
北大核心
2016年第9期1194-1203,共10页
Chinese Journal of Biotechnology
基金
国家自然科学基金(No.31402174)
河北省高等学校专业综合改革项目
河北省教育厅重点项目(No.ZD2015040)资助~~
关键词
肥大细胞
甘露糖受体
口蹄疫病毒
天然免疫
差异表达
mast cells
mannose receptor
foot-and-mouth disease virus
innate immunity
differential expression
