Clonorchis sinensis MF6p/HDM (CsMF6p/HDM) induces pro-inflammatory immune response in RAW 264.7 macrophage cells via NF-κB-dependent MAPK pathways
Abstract Background MF6p/host defense molecules (HDMs) are a broad family of small proteins secreted by helminth parasites. Although the physiological role of MF6p/HDMs in trematode parasites is not fully understood, their potential biological function in maintaining heme homeostasis and modulating...
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| Format: | Article |
| Language: | English |
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BMC
2020-01-01
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| Series: | Parasites & Vectors |
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| Online Access: | https://doi.org/10.1186/s13071-020-3882-0 |
| _version_ | 1818595668560707584 |
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| author | Jung-Mi Kang Won Gi Yoo Hương Giang Lê Jinyoung Lee Woon-Mok Sohn Byoung-Kuk Na |
| author_facet | Jung-Mi Kang Won Gi Yoo Hương Giang Lê Jinyoung Lee Woon-Mok Sohn Byoung-Kuk Na |
| author_sort | Jung-Mi Kang |
| collection | DOAJ |
| description | Abstract Background MF6p/host defense molecules (HDMs) are a broad family of small proteins secreted by helminth parasites. Although the physiological role of MF6p/HDMs in trematode parasites is not fully understood, their potential biological function in maintaining heme homeostasis and modulating host immune response has been proposed. Methods A gene encoding the MF6p/HDM of Clonorchis sinensis (CsMF6p/HDM) was cloned. Recombinant CsMF6p/HDM (rCsMF6p/HDM) was expressed in Escherichia coli. The biochemical and immunological properties of rCsMF6/HDM were analyzed. CsMF6p/HDM induced pro-inflammatory response in RAW 264.7 cells was analyzed by cytokine array assay, reverse transcription polymerase chain reaction, and enzyme-linked immunosorbent assay. The structural feature of CsMF6p/HDM was analyzed by three-dimensional modeling and molecular docking simulations. Results The CsMF6p/HDM shares a high level of amino acid sequence similarity with orthologs from other trematodes and is expressed in diverse developmental stages of the parasite. The rCsMF6p/HDM bound to bacteria-derived lipopolysaccharide (LPS), without effectively neutralizing LPS-induced inflammatory response in RAW 264.7 macrophage cells. Rather, the rCsMF6p/HDM induced pro-inflammatory immune response, which is characterized by the expression of TNF-α and IL-6, in RAW 264.7 cells. The rCsMF6p/HDM-induced pro-inflammatory immune response was regulated by JNK and p38 MAPKs, and was effectively down-regulated via inhibition of NF-κB. The structural analysis of CsMF6p/HDM and the docking simulation with LPS suggested insufficient capture of LPS by CsMF6p/HDM, which suggested that rCsMF6p/HDM could not effectively neutralize LPS-induced inflammatory response in RAW 264.7 cells. Conclusions Although rCsMF6p/HDM binds to LPS, the binding affinity may not be sufficient to maintain a stable complex of rCsMF6p/HDM and LPS. Moreover, the rCsMF6p/HDM-induced pro-inflammatory response is characterized by the release of IL-6 and TNF-α in RAW 264.7 macrophage cells. The pro-inflammatory response induced by rCsMF6p/HDM is mediated via NF-κB-dependent MAPK signaling pathway. These results collectively suggest that CsMF6p/HDM mediates C. sinensis-induced inflammation cascades that eventually lead to hepatobiliary diseases. |
| first_indexed | 2024-12-16T11:19:41Z |
| format | Article |
| id | doaj.art-dd9563afa72845068db6ea809ace9a4f |
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| language | English |
| last_indexed | 2024-12-16T11:19:41Z |
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| spelling | doaj.art-dd9563afa72845068db6ea809ace9a4f2022-12-21T22:33:30ZengBMCParasites & Vectors1756-33052020-01-0113111310.1186/s13071-020-3882-0Clonorchis sinensis MF6p/HDM (CsMF6p/HDM) induces pro-inflammatory immune response in RAW 264.7 macrophage cells via NF-κB-dependent MAPK pathwaysJung-Mi Kang0Won Gi Yoo1Hương Giang Lê2Jinyoung Lee3Woon-Mok Sohn4Byoung-Kuk Na5Department of Parasitology and Tropical Medicine, and Institute of Health Sciences, Gyeongsang National University College of MedicineDepartment of Medical Environmental Biology, Chung-Ang University College of MedicineDepartment of Parasitology and Tropical Medicine, and Institute of Health Sciences, Gyeongsang National University College of MedicineDepartment of Parasitology and Tropical Medicine, and Institute of Health Sciences, Gyeongsang National University College of MedicineDepartment of Parasitology and Tropical Medicine, and Institute of Health Sciences, Gyeongsang National University College of MedicineDepartment of Parasitology and Tropical Medicine, and Institute of Health Sciences, Gyeongsang National University College of MedicineAbstract Background MF6p/host defense molecules (HDMs) are a broad family of small proteins secreted by helminth parasites. Although the physiological role of MF6p/HDMs in trematode parasites is not fully understood, their potential biological function in maintaining heme homeostasis and modulating host immune response has been proposed. Methods A gene encoding the MF6p/HDM of Clonorchis sinensis (CsMF6p/HDM) was cloned. Recombinant CsMF6p/HDM (rCsMF6p/HDM) was expressed in Escherichia coli. The biochemical and immunological properties of rCsMF6/HDM were analyzed. CsMF6p/HDM induced pro-inflammatory response in RAW 264.7 cells was analyzed by cytokine array assay, reverse transcription polymerase chain reaction, and enzyme-linked immunosorbent assay. The structural feature of CsMF6p/HDM was analyzed by three-dimensional modeling and molecular docking simulations. Results The CsMF6p/HDM shares a high level of amino acid sequence similarity with orthologs from other trematodes and is expressed in diverse developmental stages of the parasite. The rCsMF6p/HDM bound to bacteria-derived lipopolysaccharide (LPS), without effectively neutralizing LPS-induced inflammatory response in RAW 264.7 macrophage cells. Rather, the rCsMF6p/HDM induced pro-inflammatory immune response, which is characterized by the expression of TNF-α and IL-6, in RAW 264.7 cells. The rCsMF6p/HDM-induced pro-inflammatory immune response was regulated by JNK and p38 MAPKs, and was effectively down-regulated via inhibition of NF-κB. The structural analysis of CsMF6p/HDM and the docking simulation with LPS suggested insufficient capture of LPS by CsMF6p/HDM, which suggested that rCsMF6p/HDM could not effectively neutralize LPS-induced inflammatory response in RAW 264.7 cells. Conclusions Although rCsMF6p/HDM binds to LPS, the binding affinity may not be sufficient to maintain a stable complex of rCsMF6p/HDM and LPS. Moreover, the rCsMF6p/HDM-induced pro-inflammatory response is characterized by the release of IL-6 and TNF-α in RAW 264.7 macrophage cells. The pro-inflammatory response induced by rCsMF6p/HDM is mediated via NF-κB-dependent MAPK signaling pathway. These results collectively suggest that CsMF6p/HDM mediates C. sinensis-induced inflammation cascades that eventually lead to hepatobiliary diseases.https://doi.org/10.1186/s13071-020-3882-0Clonorchis sinensisMF6p/host defense moleculeLipopolysaccharidePro-inflammatory immune responseStructureDocking |
| spellingShingle | Jung-Mi Kang Won Gi Yoo Hương Giang Lê Jinyoung Lee Woon-Mok Sohn Byoung-Kuk Na Clonorchis sinensis MF6p/HDM (CsMF6p/HDM) induces pro-inflammatory immune response in RAW 264.7 macrophage cells via NF-κB-dependent MAPK pathways Parasites & Vectors Clonorchis sinensis MF6p/host defense molecule Lipopolysaccharide Pro-inflammatory immune response Structure Docking |
| title | Clonorchis sinensis MF6p/HDM (CsMF6p/HDM) induces pro-inflammatory immune response in RAW 264.7 macrophage cells via NF-κB-dependent MAPK pathways |
| title_full | Clonorchis sinensis MF6p/HDM (CsMF6p/HDM) induces pro-inflammatory immune response in RAW 264.7 macrophage cells via NF-κB-dependent MAPK pathways |
| title_fullStr | Clonorchis sinensis MF6p/HDM (CsMF6p/HDM) induces pro-inflammatory immune response in RAW 264.7 macrophage cells via NF-κB-dependent MAPK pathways |
| title_full_unstemmed | Clonorchis sinensis MF6p/HDM (CsMF6p/HDM) induces pro-inflammatory immune response in RAW 264.7 macrophage cells via NF-κB-dependent MAPK pathways |
| title_short | Clonorchis sinensis MF6p/HDM (CsMF6p/HDM) induces pro-inflammatory immune response in RAW 264.7 macrophage cells via NF-κB-dependent MAPK pathways |
| title_sort | clonorchis sinensis mf6p hdm csmf6p hdm induces pro inflammatory immune response in raw 264 7 macrophage cells via nf κb dependent mapk pathways |
| topic | Clonorchis sinensis MF6p/host defense molecule Lipopolysaccharide Pro-inflammatory immune response Structure Docking |
| url | https://doi.org/10.1186/s13071-020-3882-0 |
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