Wen YuLing, Wang YuJian, Wang WenJuan, Lu MingMin, Ehsan Muhammad, Tian XiaoWei, Yan RuoFeng, Song XiaoKai, Xu LiXin, Li XiangRui
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, 210095, PR China.
Vet Parasitol. 2017 Aug 30;243:100-104. doi: 10.1016/j.vetpar.2017.06.018. Epub 2017 Jun 23.
In our previous proteomics study, we identified Miro domain-containing protein (Miro-1), an excretory and secretory product of the pole worm, Haemonchus contortus, binds to goat peripheral blood mononuclear cells (PBMCs) in vivo. However, our understanding of the role of Miro-1and its potential immune impact on goat PBMCs is limited. The aim of the present study was to evaluate the effects of Miro-1 on functions of goat PBMCs in vitro. Recombinant protein (rMiro-1) was expressed in a prokaryote and incubated with goat PBMCs. Western blot analysis showed that rMiro-1 is successfully recognized by goat sera infected with H. contortus. Immunofluorescence analysis using rat antibodies against rMiro-1 indicated that this protein binds to goat PBMCs in vitro. Treatment of goat PBMCs/monocytes with various concentrations of rMiro-1 resulted in the upregulation of IL-2, IL-4, and IL-17, which in turn promoted cell proliferation, migration, the release of NO in PBMCs, and enhancement of phagocytosis of monocytes. These findings suggested that rMiro-1 stimulates PBMCs activity.
在我们之前的蛋白质组学研究中,我们鉴定出了含米罗结构域蛋白(Miro-1),捻转血矛线虫的一种排泄和分泌产物,它在体内可与山羊外周血单个核细胞(PBMCs)结合。然而,我们对Miro-1的作用及其对山羊PBMCs潜在免疫影响的了解有限。本研究的目的是在体外评估Miro-1对山羊PBMCs功能的影响。重组蛋白(rMiro-1)在原核生物中表达,并与山羊PBMCs孵育。蛋白质免疫印迹分析表明,rMiro-1能被感染捻转血矛线虫的山羊血清成功识别。使用抗rMiro-1的大鼠抗体进行的免疫荧光分析表明,该蛋白在体外可与山羊PBMCs结合。用不同浓度的rMiro-1处理山羊PBMCs/单核细胞会导致IL-2、IL-4和IL-17上调,进而促进细胞增殖、迁移、PBMCs中NO的释放以及单核细胞吞噬作用的增强。这些发现表明rMiro-1可刺激PBMCs活性。