Identification of antigens in the Trichinella spiralis extracellular vesicles for serological detection of early stage infection in swine

作者：Cheng-Yao Li, Chen Li, Fengyan Xu, Haolu Wang, Xuemin Jin, Yuanyuan Zhang, Xiaolei Liu, Ruizhe Wang, Xihuo You, Mingyuan Liu, Xue Bai, Yong Yang · 发表于：Parasites & Vectors · 年份：2023 · DOI：10.1186/s13071-023-06013-7 · 被引用次数：9 · 研究领域：Parasitic Diseases Research and Treatment、Helminth infection and control、Bacterial Infections and Vaccines

BACKGROUND: Several studies have reported the roles of Trichinella spiralis extracellular vesicles in immune regulation and pathogen diagnosis. Currently, the T. spiralis muscle larvae excretory/secretory product (Ts-ML-ES) is the antigen recommended by the International Commission on Trichinellosis (ICT) for serological diagnosis of trichinellosis. However, it can only be used to detect middle and late stages of infections, and cross-reactions with other parasite detections occur. Therefore, there is a need to identify antigens for specific detection of early stage trichinellosis. METHODS: Extracellular vesicles of T. spiralis muscle larvae (Ts-ML-EVs) were isolated by ultracentrifugation and characterized by transmission electron microscopy, nanoparticle tracking analysis, flow cytometry and western blot. Ts-ML-EVs protein profiles were analyzed by LC-MS/MS proteomics for identification of potential antigens (Ts-TTPA). Ts-TTPA were cloned into pMAL-c5X vector and expressed as recombinant proteins for evaluation of potential as detected antigens by western blot and ELISA. RESULTS: Isolated Ts-ML-EVs were round or elliptic (with diameters between 110.1 and 307.6 nm), showing a bilayer membrane structure. The specific surface markers on the Ts-ML-EVs were CD81, CD63, enolase and the 14-3-3 protein. A total of 53 proteins were identified by LC-MS/MS, including a variety of molecules that have been reported as potential detection and vaccine candidates. The cDNA of Ts-TTPA selec...