Identification of antigens in the Trichinella spiralis extracellular vesicles for serological detection of early stage infection in swine
Abstract 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 Trichi...
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| Format: | Article |
| Language: | English |
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BMC
2023-10-01
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| Series: | Parasites & Vectors |
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| Online Access: | https://doi.org/10.1186/s13071-023-06013-7 |
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| author | Chengyao Li Chen Li Fengyan Xu Haolu Wang Xuemin Jin Yuanyuan Zhang Xiaolei Liu Ruizhe Wang Xihuo You Mingyuan Liu Xue Bai Yong Yang |
| author_facet | Chengyao Li Chen Li Fengyan Xu Haolu Wang Xuemin Jin Yuanyuan Zhang Xiaolei Liu Ruizhe Wang Xihuo You Mingyuan Liu Xue Bai Yong Yang |
| author_sort | Chengyao Li |
| collection | DOAJ |
| description | Abstract 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 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 antigen could 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. Conclusions This 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 |
| first_indexed | 2024-03-11T15:15:38Z |
| format | Article |
| id | doaj.art-3667f8451ccb48339e9f2a32de1c55e0 |
| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-03-11T15:15:38Z |
| publishDate | 2023-10-01 |
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| series | Parasites & Vectors |
| spelling | doaj.art-3667f8451ccb48339e9f2a32de1c55e02023-10-29T12:16:18ZengBMCParasites & Vectors1756-33052023-10-0116111210.1186/s13071-023-06013-7Identification of antigens in the Trichinella spiralis extracellular vesicles for serological detection of early stage infection in swineChengyao Li0Chen Li1Fengyan Xu2Haolu Wang3Xuemin Jin4Yuanyuan Zhang5Xiaolei Liu6Ruizhe Wang7Xihuo You8Mingyuan Liu9Xue Bai10Yong Yang11State 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 UniversityState 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 UniversityState 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 UniversityState 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 UniversityState 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 UniversityState 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 UniversityState 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 UniversityState 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 UniversityBeijing Agrichina Pharmaceutical Co., LtdState 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 UniversityState 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 UniversityState 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 UniversityAbstract 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 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 antigen could 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. Conclusions This 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 Abstracthttps://doi.org/10.1186/s13071-023-06013-7Trichinella spiralisDetection antigens of early stageProteomicsExtracellular vesicle |
| spellingShingle | Chengyao Li Chen Li Fengyan Xu Haolu Wang Xuemin Jin Yuanyuan Zhang Xiaolei Liu Ruizhe Wang Xihuo You Mingyuan Liu Xue Bai Yong Yang Identification of antigens in the Trichinella spiralis extracellular vesicles for serological detection of early stage infection in swine Parasites & Vectors Trichinella spiralis Detection antigens of early stage Proteomics Extracellular vesicle |
| title | Identification of antigens in the Trichinella spiralis extracellular vesicles for serological detection of early stage infection in swine |
| title_full | Identification of antigens in the Trichinella spiralis extracellular vesicles for serological detection of early stage infection in swine |
| title_fullStr | Identification of antigens in the Trichinella spiralis extracellular vesicles for serological detection of early stage infection in swine |
| title_full_unstemmed | Identification of antigens in the Trichinella spiralis extracellular vesicles for serological detection of early stage infection in swine |
| title_short | Identification of antigens in the Trichinella spiralis extracellular vesicles for serological detection of early stage infection in swine |
| title_sort | identification of antigens in the trichinella spiralis extracellular vesicles for serological detection of early stage infection in swine |
| topic | Trichinella spiralis Detection antigens of early stage Proteomics Extracellular vesicle |
| url | https://doi.org/10.1186/s13071-023-06013-7 |
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