NADH oxidase of Mycoplasma synoviae is a potential diagnostic antigen, plasminogen/fibronectin binding protein and a putative adhesin
Abstract Background Mycoplasma synoviae (MS) is an important pathogen causing respiratory diseases and arthritis in chickens and turkeys, thus, resulting in serious economic losses to the poultry industry. Membrane-associated proteins are thought to play important roles in cytoadherence and pathogen...
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| Format: | Article |
| Language: | English |
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BMC
2022-12-01
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| Series: | BMC Veterinary Research |
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| Online Access: | https://doi.org/10.1186/s12917-022-03556-2 |
| _version_ | 1797973546904846336 |
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| author | Zengjin Hu Haoran Li Yuxin Zhao Guijun Wang Yuanbing Shang Yuetong Chen Shaohui Wang Mingxing Tian Jingjing Qi Shengqing Yu |
| author_facet | Zengjin Hu Haoran Li Yuxin Zhao Guijun Wang Yuanbing Shang Yuetong Chen Shaohui Wang Mingxing Tian Jingjing Qi Shengqing Yu |
| author_sort | Zengjin Hu |
| collection | DOAJ |
| description | Abstract Background Mycoplasma synoviae (MS) is an important pathogen causing respiratory diseases and arthritis in chickens and turkeys, thus, resulting in serious economic losses to the poultry industry. Membrane-associated proteins are thought to play important roles in cytoadherence and pathogenesis. NADH oxidase (NOX) is an oxidoreductase involved in glycolysis, which is thought to be a multifunctional protein and potential virulence factor in some pathogens. However, little is known regarding the NOX of MS (MSNOX). We previously demonstrated that MSNOX was a metabolic enzyme distributed in not only the cytoplasm but also the MS membrane. This study was aimed at exploring NOX’s potential as a diagnostic antigen and its role in MS cytoadherence. Results Western blots and ELISAs indicated that recombinant MSNOX (rMSNOX) protein reacted with sera positive for various MS isolates, but not MG isolates or other avian pathogens, thus, suggesting that rMSNOX is a potential diagnostic antigen. In addition, rabbit anti-rMSNOX serum showed substantial complement-dependent mycoplasmacidal activity toward various MS isolates and MG Rlow. MSNOX protein was found not only in the cytoplasm but also on the membrane of MS through suspension immunofluorescence and immunogold electron microscopy assays. Indirect immunofluorescence assays indicated that rMSNOX adhered to DF-1 cells, and this adherence was inhibited by rabbit anti-rMSNOX, but not anti-MG serum. Furthermore, indirect immunofluorescence and colony counting assays confirmed that the rabbit anti-rMSNOX serum inhibited the adherence of various MS isolates but not MG Rlow to DF-1 cells. Moreover, plasminogen (Plg)- and fibronectin (Fn)-binding assays demonstrated that rMSNOX bound Plg and Fn in a dose-dependent manner, thereby further confirming that MSNOX may be a putative adhesin. Conclusion MSNOX was identified to be a surface immunogenic protein that has good immunoreactivity and specificity in Western blot and ELISA, and therefore, may be used as a potential diagnostic antigen in the future. In addition, rMSNOX adhered to DF-1 cells, an effect inhibited by rabbit anti-rMSNOX, but not anti-MG serum, and anti-rMSNOX serum inhibited the adherence of various MS isolates, but not MG Rlow, to DF-1 cells, thus indicating that the inhibition of adherence by anti-MSNOX serum was MS specific. Moreover, rMSNOX adhered to extracellular matrix proteins including Plg and Fn, thus suggesting that NOX may play important roles in MS cytoadherence and pathogenesis. Besides, rabbit anti-rMSNOX serum presented complement-dependent mycoplasmacidal activity toward both MS and MG, indicating the MSNOX may be further studied as a potential protective vaccine candidate. |
| first_indexed | 2024-04-11T04:05:59Z |
| format | Article |
| id | doaj.art-1be4d8b669a6405e89b43530a774c6a2 |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-04-11T04:05:59Z |
| publishDate | 2022-12-01 |
| publisher | BMC |
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| series | BMC Veterinary Research |
| spelling | doaj.art-1be4d8b669a6405e89b43530a774c6a22023-01-01T12:21:42ZengBMCBMC Veterinary Research1746-61482022-12-0118111410.1186/s12917-022-03556-2NADH oxidase of Mycoplasma synoviae is a potential diagnostic antigen, plasminogen/fibronectin binding protein and a putative adhesinZengjin Hu0Haoran Li1Yuxin Zhao2Guijun Wang3Yuanbing Shang4Yuetong Chen5Shaohui Wang6Mingxing Tian7Jingjing Qi8Shengqing Yu9Shanghai Veterinary Research Institute, the Chinese Academy of Agricultural Sciences (CAAS)Shanghai Veterinary Research Institute, the Chinese Academy of Agricultural Sciences (CAAS)Shanghai Veterinary Research Institute, the Chinese Academy of Agricultural Sciences (CAAS)College of Animal Science and Technology, Anhui Agricultural UniversityShanghai Veterinary Research Institute, the Chinese Academy of Agricultural Sciences (CAAS)Shanghai Veterinary Research Institute, the Chinese Academy of Agricultural Sciences (CAAS)Shanghai Veterinary Research Institute, the Chinese Academy of Agricultural Sciences (CAAS)Shanghai Veterinary Research Institute, the Chinese Academy of Agricultural Sciences (CAAS)Shanghai Veterinary Research Institute, the Chinese Academy of Agricultural Sciences (CAAS)Shanghai Veterinary Research Institute, the Chinese Academy of Agricultural Sciences (CAAS)Abstract Background Mycoplasma synoviae (MS) is an important pathogen causing respiratory diseases and arthritis in chickens and turkeys, thus, resulting in serious economic losses to the poultry industry. Membrane-associated proteins are thought to play important roles in cytoadherence and pathogenesis. NADH oxidase (NOX) is an oxidoreductase involved in glycolysis, which is thought to be a multifunctional protein and potential virulence factor in some pathogens. However, little is known regarding the NOX of MS (MSNOX). We previously demonstrated that MSNOX was a metabolic enzyme distributed in not only the cytoplasm but also the MS membrane. This study was aimed at exploring NOX’s potential as a diagnostic antigen and its role in MS cytoadherence. Results Western blots and ELISAs indicated that recombinant MSNOX (rMSNOX) protein reacted with sera positive for various MS isolates, but not MG isolates or other avian pathogens, thus, suggesting that rMSNOX is a potential diagnostic antigen. In addition, rabbit anti-rMSNOX serum showed substantial complement-dependent mycoplasmacidal activity toward various MS isolates and MG Rlow. MSNOX protein was found not only in the cytoplasm but also on the membrane of MS through suspension immunofluorescence and immunogold electron microscopy assays. Indirect immunofluorescence assays indicated that rMSNOX adhered to DF-1 cells, and this adherence was inhibited by rabbit anti-rMSNOX, but not anti-MG serum. Furthermore, indirect immunofluorescence and colony counting assays confirmed that the rabbit anti-rMSNOX serum inhibited the adherence of various MS isolates but not MG Rlow to DF-1 cells. Moreover, plasminogen (Plg)- and fibronectin (Fn)-binding assays demonstrated that rMSNOX bound Plg and Fn in a dose-dependent manner, thereby further confirming that MSNOX may be a putative adhesin. Conclusion MSNOX was identified to be a surface immunogenic protein that has good immunoreactivity and specificity in Western blot and ELISA, and therefore, may be used as a potential diagnostic antigen in the future. In addition, rMSNOX adhered to DF-1 cells, an effect inhibited by rabbit anti-rMSNOX, but not anti-MG serum, and anti-rMSNOX serum inhibited the adherence of various MS isolates, but not MG Rlow, to DF-1 cells, thus indicating that the inhibition of adherence by anti-MSNOX serum was MS specific. Moreover, rMSNOX adhered to extracellular matrix proteins including Plg and Fn, thus suggesting that NOX may play important roles in MS cytoadherence and pathogenesis. Besides, rabbit anti-rMSNOX serum presented complement-dependent mycoplasmacidal activity toward both MS and MG, indicating the MSNOX may be further studied as a potential protective vaccine candidate.https://doi.org/10.1186/s12917-022-03556-2Mycoplasma synoviaeNADH oxidaseMembrane-localizationDiagnostic antigenCytoadherencePlasminogen/fibronectin-binding |
| spellingShingle | Zengjin Hu Haoran Li Yuxin Zhao Guijun Wang Yuanbing Shang Yuetong Chen Shaohui Wang Mingxing Tian Jingjing Qi Shengqing Yu NADH oxidase of Mycoplasma synoviae is a potential diagnostic antigen, plasminogen/fibronectin binding protein and a putative adhesin BMC Veterinary Research Mycoplasma synoviae NADH oxidase Membrane-localization Diagnostic antigen Cytoadherence Plasminogen/fibronectin-binding |
| title | NADH oxidase of Mycoplasma synoviae is a potential diagnostic antigen, plasminogen/fibronectin binding protein and a putative adhesin |
| title_full | NADH oxidase of Mycoplasma synoviae is a potential diagnostic antigen, plasminogen/fibronectin binding protein and a putative adhesin |
| title_fullStr | NADH oxidase of Mycoplasma synoviae is a potential diagnostic antigen, plasminogen/fibronectin binding protein and a putative adhesin |
| title_full_unstemmed | NADH oxidase of Mycoplasma synoviae is a potential diagnostic antigen, plasminogen/fibronectin binding protein and a putative adhesin |
| title_short | NADH oxidase of Mycoplasma synoviae is a potential diagnostic antigen, plasminogen/fibronectin binding protein and a putative adhesin |
| title_sort | nadh oxidase of mycoplasma synoviae is a potential diagnostic antigen plasminogen fibronectin binding protein and a putative adhesin |
| topic | Mycoplasma synoviae NADH oxidase Membrane-localization Diagnostic antigen Cytoadherence Plasminogen/fibronectin-binding |
| url | https://doi.org/10.1186/s12917-022-03556-2 |
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