Abstract：
Cystic echinococcosis (CE) is a zoonotic disease, caused by Echinococcus granulosus sensu lato (E. granulosus s. l.), which posed significant public health concern globally. E. granulosus s. l. annexin B18 (EgANXB18) acts as a secretory protein, exerting a crucial influence in mediating host-parasite interactions. Recombinant annexin B18 (rEgANXB18) was expressed by Escherichia coli and the immunoreactivity was assessed by western blotting. The binding affinity between rEgANXB18 and total protein of RAW264.7 cells was assessed by ELISA. The impact of rEgANXB18 on the metabolic activity of RAW264.7 cells was assayed by Cell Counting Kit-8 assay. The mRNA levels of polarization markers (inducible nitrous oxide synthase (iNOS) and arginase 1 (Arg1)) and key cellular factors (IL-1β，IL-6，IL-10 and TNFα) were evaluated by qRT-PCR. rEgANXB18 was successfully expressed and recognized by E. granulosus s.l. infected canine sera, as well as could bind to the total protein of RAW264.7 cells. Additionally, rEgANXB18 could promote metabolic activity at 5, 10, 20, and 40 μg/mL while no significant impact on metabolic activity was observed at 80 μg/mL. Co-culture RAW264.7 cells with rEgANXB18 resulted in significantly upregulation of the transcript levels of polarization markers iNOS and Arg1. Moreover, rEgANXB18 significantly upregulated the transcript levels of IL-1β, IL-6, TNFα, and IL-10, while dose-effect relationship was observed in IL-1β, IL-6, and IL-10. Our results indicated that EgANXB18 showed the potential to regulate immune response of macrophages by shifting the cell polarization and cytokine profile, thereby promoting the parasitism of CE.
摘要：
囊性包虫病（CE）是一种人畜共患疾病，由细粒棘球蚴引起（E.granulosuss.l.)，这在全球范围内引起了重大的公共卫生关注。E.granulosuss.l.膜联蛋白B18（EgANXB18）作为分泌蛋白，在介导宿主-寄生虫相互作用中发挥关键作用。重组膜联蛋白B18（rEgANXB18）由大肠杆菌表达，并通过蛋白质印迹法评估免疫反应性。通过ELISA评估rEgANXB18与RAW264.7细胞的总蛋白之间的结合亲和力。通过细胞计数试剂盒-8测定法测定rEgANXB18对RAW264.7细胞的代谢活性的影响。qRT-PCR检测极化标记物（诱导型一氧化氮合酶（iNOS）和精氨酸酶1（Arg1））和关键细胞因子（IL-1β，IL-6，IL-10和TNFα）的mRNA水平。rEgANXB18成功表达并被S.I.感染的犬血清识别，以及可以结合RAW264.7细胞的总蛋白。此外,rEgANXB18在5、10、20和40μg/mL时可促进代谢活性，而在80μg/mL时对代谢活性无明显影响。RAW264.7细胞与rEgANXB18共培养导致极化标记iNOS和Arg1的转录水平显着上调。此外，rEgANXB18显著上调IL-1β的转录水平，IL-6,TNFα,和IL-10，而在IL-1β中观察到剂量效应关系，IL-6和IL-10。我们的结果表明，EgANXB18显示出通过改变细胞极化和细胞因子谱来调节巨噬细胞免疫反应的潜力，从而促进了CE的寄生。
