Table_1_Clonorchis sinensis calcium-binding protein Cs16 causes acute hepatic injury possibly by reprogramming the metabolic pathway of bone marrow-derived monocytes.xlsx

IntroductionClonorchis sinensis infection results in various complications in the liver and biliary systems and is a neglected tropical disease in Eastern Asia. In this study, we report that C. sinensis calcium-binding protein Cs16 activates host immune cells and induces immunopathology in liver.
MethodsImmunohistochemistry was used to detect the localization of Cs16 in C. sinensis adult worms. ELISA was used to detect the serum levels of anti-Cs16 IgG antibody in infected humans and mice. Bile duct injection model was used to figure out the role of Cs16 in vivo. RT-qPCR and ELISA were used to detect the cytokine production from Cs16-treated BMMs in vitro. Seahorse assay was used to detect the metabolic pathway of Cs16-treated BMMs in vitro.
ResultCs16 localizes in the tegument and gut of C. sinensis. Humans and mice with C. sinensis infection exhibited increased levels of anti-Cs16-specific antibody. Using the bile duct injection technique, we found that Cs16 induced obvious inflammation and hepatic necrosis in vivo. Cs16 treatment caused the upregulation of inflammatory cytokines in innate immune cells. Moreover, Cs16-treated monocytes relied more on the glycolytic metabolic pathway.
DiscussionOur findings suggest that Cs16 is a potential pathogenic factor derived from C. sinensis adult worm. By reprogramming the metabolic pathway of innate immune cells, Cs16 triggers pro-inflammatory responses in the liver, and therefore, Cs16 is a potential target for the prevention and treatment of clonorchiasis.

引言 华支睾吸虫（Clonorchis sinensis）感染可引发肝脏与胆道系统的多种并发症，是东亚地区的被忽视热带病。本研究证实，华支睾吸虫钙结合蛋白Cs16能够激活宿主免疫细胞，并诱导肝脏产生免疫病理损伤。
方法 采用免疫组织化学（Immunohistochemistry）法检测Cs16在华支睾吸虫成虫中的组织定位；通过酶联免疫吸附实验（ELISA）检测感染华支睾吸虫的人类及小鼠血清中抗Cs16 IgG抗体水平；构建胆道注射模型以明确Cs16在体内的生物学作用；采用实时定量聚合酶链式反应（RT-qPCR）与ELISA检测体外经Cs16处理的骨髓来源巨噬细胞（BMMs）的细胞因子分泌情况；利用海马代谢分析仪（Seahorse assay）检测体外经Cs16处理的BMMs的代谢通路变化。
结果 Cs16定位于华支睾吸虫的体被与肠道。感染华支睾吸虫的人类与小鼠体内抗Cs16特异性抗体水平显著升高。通过胆道注射实验，我们发现Cs16在体内可诱导明显的炎症反应与肝脏坏死。经Cs16处理的先天免疫细胞中炎症细胞因子表达上调。此外，经Cs16处理的单核细胞更依赖糖酵解代谢通路。
讨论 本研究结果表明，Cs16是源自华支睾吸虫成虫的潜在致病因子。通过重编程先天免疫细胞的代谢通路，Cs16可触发肝脏的促炎反应，因此Cs16有望成为华支睾吸虫病防治的潜在靶点。