He Peixia, Zhang Dejia, Wang Mengqi, Duan Rui, Zhao Yuyuan, Wang Sirui, Yang Xing, Liu Xiaolei, Sun Shumin
College of Veterinary Medicine, Yunnan Agricultural University, Kunming, China.
State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, Institute of Zoonosis, and College of Veterinary Medicine, Jilin University, Changchun, China.
PLoS Negl Trop Dis. 2025 Apr 17;19(4):e0013015. doi: 10.1371/journal.pntd.0013015. eCollection 2025 Apr.
Cysticercosis is a severe foodborne zoonotic parasitosis infected by the metacestode larvae of Taenia solium. However, its invasion mechanism is still not clear, which might provide the important evidence for treatment or vaccine. It was reported the annexin involved in the physiological and pathological functions of Cysticercus cellulosae. However, the regulatory mechanisms and roles of annexin B1 and annexin B2 in the invasion and immune escape of Cysticercus cellulosae have not been fully explored.
The annexin was acquired by cloning in prokaryotic expression vector, expressed in Escherichia coli, and purified by affinity chromatography. Its expression was determined by immunohistochemistry. The anticoagulant function and its underlying mechanism was verified by the determination of activated partial thromboplastin time, prothrombin time and phospholipid binding activity. The membrane repair function was verified by cell culture, transfection, and laser confocal technology.
Immunohistochemistry results showed the B1 and B2 were mainly expressed on the body surface and the surface of digestive glands of Cysticercus cellulosae. The Blood coagulation results illustrated the B1 and B2 can prolong the time of both exogenous and endogenous coagulation pathways, with B2 having a more significant effect. They tend to bind to phosphatidylserine, possibly interfering with coagulation complex formation and inhibiting the coagulation pathway, and may assist in the worm's penetration through blood vessels and migration to parasitic sites. The plasma membrane repair test revealed the cells transfected with B1 and B2 genes have a significantly shorter plasma membrane repair time than the control group, suggesting that these proteins may be involved in repairing the worm's body surface to resist the immune system's attack when the host immune system attacks.
The Annexin B1 and Annexin B2 of Cysticercus cellulosae possess anticoagulant properties and can assist in membrane repair. Given these functions, it is speculated that they play a crucial role in immune evasion and invasion. However, further experiments are required to provide direct evidence to further validate these speculations.
囊尾蚴病是一种严重的食源性人畜共患寄生虫病，由猪带绦虫的中绦期幼虫感染引起。然而，其入侵机制仍不清楚，这可能为治疗或疫苗提供重要依据。有报道称膜联蛋白参与了猪囊尾蚴的生理和病理功能。然而，膜联蛋白B1和膜联蛋白B2在猪囊尾蚴入侵和免疫逃逸中的调控机制及作用尚未得到充分研究。
通过克隆将膜联蛋白构建到原核表达载体中，在大肠杆菌中表达，并通过亲和层析进行纯化。通过免疫组织化学确定其表达情况。通过测定活化部分凝血活酶时间、凝血酶原时间和磷脂结合活性来验证其抗凝功能及其潜在机制。通过细胞培养、转染和激光共聚焦技术验证其膜修复功能。
免疫组织化学结果显示，B1和B2主要在猪囊尾蚴的体表和消化腺表面表达。凝血结果表明，B1和B2均可延长外源性和内源性凝血途径的时间，其中B2的作用更显著。它们倾向于与磷脂酰丝氨酸结合，可能干扰凝血复合物的形成并抑制凝血途径，还可能协助虫体穿透血管并迁移至寄生部位。质膜修复试验表明，转染B1和B2基因细胞的质膜修复时间明显短于对照组，提示这些蛋白可能在宿主免疫系统攻击时参与修复虫体体表以抵抗免疫系统的攻击。
猪囊尾蚴的膜联蛋白B1和膜联蛋白B2具有抗凝特性并可协助膜修复。鉴于这些功能，推测它们在免疫逃避和入侵中起关键作用。然而，需要进一步的实验提供直接证据以进一步验证这些推测。
