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

Parasites & Vectors

Research

Xihuo You¹  Xiaolei Liu²  Ruizhe Wang²  Xue Bai²  Haolu Wang²  Chengyao Li²  Xuemin Jin²  Yuanyuan Zhang²  Chen Li²  Fengyan Xu²  Mingyuan Liu³  Yong Yang⁴ 

[1] Beijing Agrichina Pharmaceutical Co., Ltd, Wangzhuang Industrial Park, Airport Road, Shahe, Changping District, Beijing, China;State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of Ministry of Education, Institute of Zoonosis, and College of Veterinary Medicine, Jilin University, Changchun, China;State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of Ministry of Education, Institute of Zoonosis, and College of Veterinary Medicine, Jilin University, Changchun, China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, People’s Republic of China;State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of Ministry of Education, Institute of Zoonosis, and College of Veterinary Medicine, Jilin University, Changchun, China;School of Basic Medical Science, Shan Xi Medical University, Taiyuan, China;

关键词: Trichinella spiralis;    Detection antigens of early stage;    Proteomics;    Extracellular vesicle;   

DOI  :  10.1186/s13071-023-06013-7

 received in 2023-07-18, accepted in 2023-10-14,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundSeveral 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.MethodsExtracellular 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.ResultsIsolated 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 selected in this study has a total length of 1152 bp, encoding 384 amino acids with a molecular weight of 44.19 kDa. It contains a trypsin domain and can be recognized by anti-His antibody. It reacted with swine sera infected with 10,000 T. spiralis at 15, 25, 35 and 60 days post-infection (dpi). At 10 μg/ml, this antigencould detect T. spiralis antibodies from the swine sera at 13 dpi. There were no cross-reactions with the swine sera infected with other parasites including Clonorchis sinensis, Toxoplasma gondii, Taenia suis, Ascaris suis and Trichuris suis.ConclusionsThis study identifies potential early stage detection antigens and more thoroughly characterizes a serine protease domain-containing protein. Extracellular vesicle proteins may be explored as effective antigens for the early stage detection of trichinellosis.Graphical Abstract

【 授权许可】

CC BY   
© The Author(s) 2023

【 图 表 】

Fig. 1

Fig. 2

Fig. 2

Fig. 4

Fig. 3

Fig. 1

Fig. 9