Trichinella spiralis galectin binding to toll-like receptor 4 induces intestinal inflammation and mediates larval invasion of gut mucosa
Abstract Previous studies showed that Trichinella spiralis galectin (Tsgal) facilitates larval invasion of intestinal epithelium cells (IECs). However, IEC proteins binding with Tsgal were not identified, and the mechanism by which Tsgal promotes larval invasion is not clear. Toll-like receptors (TL...
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| Format: | Article |
| Language: | English |
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BMC
2023-11-01
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| Series: | Veterinary Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13567-023-01246-x |
| _version_ | 1797414268355739648 |
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| author | Kai Ning Ma Yao Zhang Zhao Yu Zhang Bo Ning Wang Yan Yan Song Lu Lu Han Xin Zhuo Zhang Shao Rong Long Jing Cui Zhong Quan Wang |
| author_facet | Kai Ning Ma Yao Zhang Zhao Yu Zhang Bo Ning Wang Yan Yan Song Lu Lu Han Xin Zhuo Zhang Shao Rong Long Jing Cui Zhong Quan Wang |
| author_sort | Kai Ning Ma |
| collection | DOAJ |
| description | Abstract Previous studies showed that Trichinella spiralis galectin (Tsgal) facilitates larval invasion of intestinal epithelium cells (IECs). However, IEC proteins binding with Tsgal were not identified, and the mechanism by which Tsgal promotes larval invasion is not clear. Toll-like receptors (TLRs) are protein receptors responsible for recognition of pathogens. The aim of this study was to investigate whether recombinant Tsgal (rTsgal) binds to TLR-4, activates inflammatory pathway in gut epithelium and mediates T. spiralis invasion. Indirect immunofluorescence (IIF), GST pull-down and co-immunoprecipitation (Co-IP) assays confirmed specific binding between rTsgal and TLR-4 in Caco-2 cells. qPCR and Western blotting showed that binding of rTsgal with TLR-4 up-regulated the TLR-4 transcription and expression in Caco-2 cells, and activated p-NF-κB p65 and p-ERK1/2. Activation of inflammatory pathway TLR-4/MAPK-NF-κB by rTsgal up-regulated pro-inflammatory cytokines (IL-1β and IL-6) and down-regulated anti-inflammatory cytokine TGF-β in Caco-2 cells, and induced intestinal inflammation. TAK-242 (TLR-4 inhibitor) and PDTC (NF-κB inhibitor) significantly inhibited the activation of TLR-4 and MAPK-NF-κB pathway. Moreover, the two inhibitors also inhibited IL-1β and IL-6 expression, and increased TGF-β expression in Caco-2 cells. In T. spiralis infected mice, the two inhibitors also inhibited the activation of TLR-4/MAPK-NF-κB pathway, ameliorated intestinal inflammation, impeded larval invasion of gut mucosa and reduced intestinal adult burdens. The results showed that rTsgal binding to TLR-4 in gut epithelium activated MAPK-NF-κB signaling pathway, induced the expression of TLR-4 and pro-inflammatory cytokines, and mediated larval invasion. Tsgal might be regarded as a candidate molecular target of vaccine against T. spiralis enteral invasive stage. |
| first_indexed | 2024-03-09T05:30:34Z |
| format | Article |
| id | doaj.art-ba5d43a06b944d64bc109f8c12d7a040 |
| institution | Directory Open Access Journal |
| issn | 1297-9716 |
| language | English |
| last_indexed | 2024-03-09T05:30:34Z |
| publishDate | 2023-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Veterinary Research |
| spelling | doaj.art-ba5d43a06b944d64bc109f8c12d7a0402023-12-03T12:33:06ZengBMCVeterinary Research1297-97162023-11-0154112110.1186/s13567-023-01246-xTrichinella spiralis galectin binding to toll-like receptor 4 induces intestinal inflammation and mediates larval invasion of gut mucosaKai Ning Ma0Yao Zhang1Zhao Yu Zhang2Bo Ning Wang3Yan Yan Song4Lu Lu Han5Xin Zhuo Zhang6Shao Rong Long7Jing Cui8Zhong Quan Wang9Department of Parasitology, Medical College, Zhengzhou UniversityDepartment of Parasitology, Medical College, Zhengzhou UniversityDepartment of Parasitology, Medical College, Zhengzhou UniversityDepartment of Parasitology, Medical College, Zhengzhou UniversityDepartment of Parasitology, Medical College, Zhengzhou UniversityDepartment of Parasitology, Medical College, Zhengzhou UniversityDepartment of Parasitology, Medical College, Zhengzhou UniversityDepartment of Parasitology, Medical College, Zhengzhou UniversityDepartment of Parasitology, Medical College, Zhengzhou UniversityDepartment of Parasitology, Medical College, Zhengzhou UniversityAbstract Previous studies showed that Trichinella spiralis galectin (Tsgal) facilitates larval invasion of intestinal epithelium cells (IECs). However, IEC proteins binding with Tsgal were not identified, and the mechanism by which Tsgal promotes larval invasion is not clear. Toll-like receptors (TLRs) are protein receptors responsible for recognition of pathogens. The aim of this study was to investigate whether recombinant Tsgal (rTsgal) binds to TLR-4, activates inflammatory pathway in gut epithelium and mediates T. spiralis invasion. Indirect immunofluorescence (IIF), GST pull-down and co-immunoprecipitation (Co-IP) assays confirmed specific binding between rTsgal and TLR-4 in Caco-2 cells. qPCR and Western blotting showed that binding of rTsgal with TLR-4 up-regulated the TLR-4 transcription and expression in Caco-2 cells, and activated p-NF-κB p65 and p-ERK1/2. Activation of inflammatory pathway TLR-4/MAPK-NF-κB by rTsgal up-regulated pro-inflammatory cytokines (IL-1β and IL-6) and down-regulated anti-inflammatory cytokine TGF-β in Caco-2 cells, and induced intestinal inflammation. TAK-242 (TLR-4 inhibitor) and PDTC (NF-κB inhibitor) significantly inhibited the activation of TLR-4 and MAPK-NF-κB pathway. Moreover, the two inhibitors also inhibited IL-1β and IL-6 expression, and increased TGF-β expression in Caco-2 cells. In T. spiralis infected mice, the two inhibitors also inhibited the activation of TLR-4/MAPK-NF-κB pathway, ameliorated intestinal inflammation, impeded larval invasion of gut mucosa and reduced intestinal adult burdens. The results showed that rTsgal binding to TLR-4 in gut epithelium activated MAPK-NF-κB signaling pathway, induced the expression of TLR-4 and pro-inflammatory cytokines, and mediated larval invasion. Tsgal might be regarded as a candidate molecular target of vaccine against T. spiralis enteral invasive stage.https://doi.org/10.1186/s13567-023-01246-xTrichinella spiralisgalectinToll-like receptor 4 (TLR-4)gut epitheliuminvasionMAPK-NF-κB pathway |
| spellingShingle | Kai Ning Ma Yao Zhang Zhao Yu Zhang Bo Ning Wang Yan Yan Song Lu Lu Han Xin Zhuo Zhang Shao Rong Long Jing Cui Zhong Quan Wang Trichinella spiralis galectin binding to toll-like receptor 4 induces intestinal inflammation and mediates larval invasion of gut mucosa Veterinary Research Trichinella spiralis galectin Toll-like receptor 4 (TLR-4) gut epithelium invasion MAPK-NF-κB pathway |
| title | Trichinella spiralis galectin binding to toll-like receptor 4 induces intestinal inflammation and mediates larval invasion of gut mucosa |
| title_full | Trichinella spiralis galectin binding to toll-like receptor 4 induces intestinal inflammation and mediates larval invasion of gut mucosa |
| title_fullStr | Trichinella spiralis galectin binding to toll-like receptor 4 induces intestinal inflammation and mediates larval invasion of gut mucosa |
| title_full_unstemmed | Trichinella spiralis galectin binding to toll-like receptor 4 induces intestinal inflammation and mediates larval invasion of gut mucosa |
| title_short | Trichinella spiralis galectin binding to toll-like receptor 4 induces intestinal inflammation and mediates larval invasion of gut mucosa |
| title_sort | trichinella spiralis galectin binding to toll like receptor 4 induces intestinal inflammation and mediates larval invasion of gut mucosa |
| topic | Trichinella spiralis galectin Toll-like receptor 4 (TLR-4) gut epithelium invasion MAPK-NF-κB pathway |
| url | https://doi.org/10.1186/s13567-023-01246-x |
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