Abstract:
Parasitic helminths modify host immune reactions to promote long-term parasitism. We previously purified a glycoprotein, plerocercoid-immunosuppressive factor (P-ISF), from the excretory/secretory products of Spirometra erinaceieuropaei plerocercoids and reported its cDNA and genomic DNA sequences. In this study, we isolated extracellular vesicles (EVs) from the excretory/secretory products of S. erinaceieuropaei plerocercoids and found that they suppressed the production of nitric oxide and the gene expression of tumor necrosis factor-α, interleukin-1β, and interleukin-6 in lipopolysaccharide-stimulated macrophages. EVs are membrane-bound vesicles 50-250 nm in diameter and are localized in the whole bodies of plerocercoids. EVs from plerocercoids encapsulate a variety of unidentified proteins and microRNAs (miRNAs), which are non-coding RNAs that play essential roles in post-transcriptional gene regulation. The miRNAs of the EVs were analyzed, and 334,137 sequencing reads were mapped to the genomes of other organisms. A total of 26 different miRNA families were identified, including miR-71, miR-10-5p, miR-223, and let-7-5p, which have been reported to have immunosuppressive effects. We confirmed that P-ISF was present in the supernatant but not in the EVs by western blotting with an anti-P-ISF antibody. These results suggest that S. erinaceieuropaei plerocercoids suppress host immunity by releasing P-ISF and EVs.
摘要:
寄生虫会改变宿主的免疫反应以促进长期的寄生虫。我们之前纯化了一种糖蛋白,plerocecoid-免疫抑制因子(P-ISF),来自Spirometra的排泄/分泌产物,并报告了其cDNA和基因组DNA序列。在这项研究中,我们从S.erinaceuropaeiplerocerids的排泄/分泌产物中分离了细胞外囊泡(EV),发现它们抑制了一氧化氮的产生和肿瘤坏死因子-α的基因表达,白细胞介素-1β,和脂多糖刺激的巨噬细胞中的白细胞介素-6。EV是直径为50-250nm的膜结合囊泡,并位于plerocercoids的整个体内。来自plerocercoids的EV封装了各种未识别的蛋白质和microRNAs(miRNAs),它们是在转录后基因调控中起重要作用的非编码RNA。分析了电动汽车的miRNA,334137个测序读数被映射到其他生物体的基因组。总共鉴定了26个不同的miRNA家族,包括miR-71,miR-10-5p,miR-223和let-7-5p,据报道具有免疫抑制作用。我们通过用抗P-ISF抗体的蛋白质印迹证实了P-ISF存在于上清液中而不存在于EV中。这些结果表明,S.erinaceuropaeiplerocerids通过释放P-ISF和EV来抑制宿主免疫。