Cystic echinococcosis (CE) is a zoonotic disease, caused by sensu lato ( s. l.), which posed significant public health concern globally. s. l. annexin B18 (ANXB18) acts as a secretory protein, exerting a crucial influence in mediating host-parasite interactions. Recombinant annexin B18 (rANXB18) was expressed by and the immunoreactivity was assessed by western blotting. The binding affinity between rANXB18 and total protein of RAW264.7 cells was assessed by ELISA. The impact of rANXB18 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. rANXB18 was successfully expressed and recognized by s.l. infected canine sera, as well as could bind to the total protein of RAW264.7 cells. Additionally, rANXB18 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 rANXB18 resulted in significantly upregulation of the transcript levels of polarization markers iNOS and Arg1. Moreover, rANXB18 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 ANXB18 showed the potential to regulate immune response of macrophages by shifting the cell polarization and cytokine profile, thereby promoting the parasitism of CE.

中文翻译：

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细粒棘球绦虫膜联蛋白 B18 对小鼠巨噬细胞的影响


囊性包虫病 (CE) 是一种人畜共患疾病，由 sensu lato (s.l.) 引起，在全球范围内引起了重大的公共卫生问题。 s。湖膜联蛋白 B18 (ANXB18) 作为一种分泌蛋白，在介导宿主与寄生虫的相互作用中发挥着至关重要的影响。表达重组膜联蛋白 B18 (rANXB18)，并通过蛋白质印迹评估免疫反应性。通过 ELISA 评估 rANXB18 与 RAW264.7 细胞总蛋白之间的结合亲和力。通过 Cell Counting Kit-8 测定法测定 rANXB18 对 RAW264.7 细胞代谢活性的影响。通过qRT-PCR评估极化标记物（诱导型一氧化二氮合酶（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。 rANXB18 成功表达并被 s.l. 识别。受感染的犬血清，并且可以与 RAW264.7 细胞的总蛋白结合。此外，rANXB18 在 5、10、20 和 40 μg/mL 浓度下可促进代谢活性，而在 80 μg/mL 浓度下未观察到对代谢活性的显着影响。将 RAW264.7 细胞与 rANXB18 共培养会导致极化标记物 iNOS 和 Arg1 的转录水平显着上调。此外，rANXB18显着上调IL-1β、IL-6、TNFα和IL-10的转录水平，同时在IL-1β、IL-6和IL-10中观察到剂量效应关系。我们的结果表明，ANXB18 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 CE 的寄生。