NADH oxidase of Mycoplasma hyopneumoniae functions as a potential mediator of virulence
Abstract Background Mycoplasma hyopneumoniae (M. hyopneumoniae) is the etiological agent of enzootic pneumonia, a highly infectious swine respiratory disease that distributed worldwide. The pathogenesis and virulence factors of M. hyopneumoniae are not fully clarified. As an important virulence fact...
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| Format: | Article |
| Language: | English |
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BMC
2022-04-01
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| Series: | BMC Veterinary Research |
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| Online Access: | https://doi.org/10.1186/s12917-022-03230-7 |
| _version_ | 1811260516957421568 |
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| author | Fei Hao Xing Xie Zhixin Feng Rong Chen Yanna Wei Jin Liu Qiyan Xiong Guoqing Shao Johnson Lin |
| author_facet | Fei Hao Xing Xie Zhixin Feng Rong Chen Yanna Wei Jin Liu Qiyan Xiong Guoqing Shao Johnson Lin |
| author_sort | Fei Hao |
| collection | DOAJ |
| description | Abstract Background Mycoplasma hyopneumoniae (M. hyopneumoniae) is the etiological agent of enzootic pneumonia, a highly infectious swine respiratory disease that distributed worldwide. The pathogenesis and virulence factors of M. hyopneumoniae are not fully clarified. As an important virulence factor of bacteria, nicotinamide adenine dinucleotide (NADH) oxidase (NOX) participates in host-pathogen interaction, however, the function of NOX involved in the pathogenesis of M. hyopneumoniae is not clear. Results In this study, significant differences in NOX transcription expression levels among different strains of M. hyopneumoniae differed in virulence were identified, suggesting that NOX may be correlated with M. hyopneumoniae virulence. The nox gene of M. hyopneumoniae was cloned and expressed in Escherichia coli, and polyclonal antibodies against recombinant NOX (rNOX) were prepared. We confirmed the enzymatic activity of rNOX based on its capacity to oxidize NADH to NAD+. Flow cytometry analysis demonstrated the surface localization of NOX, and subcellular localization analysis further demonstrated that NOX exists in both the cytoplasm and cell membrane. rNOX was depicted to mediate adhesion to immortalized porcine bronchial epithelial cells (hTERT-PBECs). Pre-neutralizing M. hyopneumoniae with anti-rNOX antibody resulted in a more than 55% reduction in the adhesion rate of high- and low-virulence M. hyopneumoniae strains to hTERT-PBECs. Moreover, a significant difference appeared in the decline in CCU50 titer between virulent (168) and virulence-attenuated (168L) strains. NOX not only recognized and interacted with host fibronectin but also induced cellular oxidative stress and apoptosis in hTERT-PBECs. The release of lactate dehydrogenase by NOX in hTERT-PBECs was positively correlated with the virulence of M. hyopneumoniae strains. Conclusions NOX is considered to be a potential virulence factor of M. hyopneumoniae and may play a significant role in mediating its pathogenesis. |
| first_indexed | 2024-04-12T18:48:32Z |
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| id | doaj.art-d5a26394b3884ed0b31c1d3184239709 |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-04-12T18:48:32Z |
| publishDate | 2022-04-01 |
| publisher | BMC |
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| series | BMC Veterinary Research |
| spelling | doaj.art-d5a26394b3884ed0b31c1d31842397092022-12-22T03:20:33ZengBMCBMC Veterinary Research1746-61482022-04-0118111410.1186/s12917-022-03230-7NADH oxidase of Mycoplasma hyopneumoniae functions as a potential mediator of virulenceFei Hao0Xing Xie1Zhixin Feng2Rong Chen3Yanna Wei4Jin Liu5Qiyan Xiong6Guoqing Shao7Johnson Lin8Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base, Ministry of Science and Technology, Key Laboratory for Veterinary Bio-Product Engineering, Ministry of Agriculture and Rural Affairs, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural SciencesJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base, Ministry of Science and Technology, Key Laboratory for Veterinary Bio-Product Engineering, Ministry of Agriculture and Rural Affairs, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural SciencesJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base, Ministry of Science and Technology, Key Laboratory for Veterinary Bio-Product Engineering, Ministry of Agriculture and Rural Affairs, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural SciencesJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base, Ministry of Science and Technology, Key Laboratory for Veterinary Bio-Product Engineering, Ministry of Agriculture and Rural Affairs, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural SciencesJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base, Ministry of Science and Technology, Key Laboratory for Veterinary Bio-Product Engineering, Ministry of Agriculture and Rural Affairs, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural SciencesHubei Key Laboratory of Animal Nutrition and Feed Science, Wuhan Polytechnic UniversityJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base, Ministry of Science and Technology, Key Laboratory for Veterinary Bio-Product Engineering, Ministry of Agriculture and Rural Affairs, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural SciencesJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base, Ministry of Science and Technology, Key Laboratory for Veterinary Bio-Product Engineering, Ministry of Agriculture and Rural Affairs, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural SciencesDiscipline of Microbiology, School of Life Sciences, College of Agriculture, Engineering and Science, University of KwaZulu-Natal (Westville campus)Abstract Background Mycoplasma hyopneumoniae (M. hyopneumoniae) is the etiological agent of enzootic pneumonia, a highly infectious swine respiratory disease that distributed worldwide. The pathogenesis and virulence factors of M. hyopneumoniae are not fully clarified. As an important virulence factor of bacteria, nicotinamide adenine dinucleotide (NADH) oxidase (NOX) participates in host-pathogen interaction, however, the function of NOX involved in the pathogenesis of M. hyopneumoniae is not clear. Results In this study, significant differences in NOX transcription expression levels among different strains of M. hyopneumoniae differed in virulence were identified, suggesting that NOX may be correlated with M. hyopneumoniae virulence. The nox gene of M. hyopneumoniae was cloned and expressed in Escherichia coli, and polyclonal antibodies against recombinant NOX (rNOX) were prepared. We confirmed the enzymatic activity of rNOX based on its capacity to oxidize NADH to NAD+. Flow cytometry analysis demonstrated the surface localization of NOX, and subcellular localization analysis further demonstrated that NOX exists in both the cytoplasm and cell membrane. rNOX was depicted to mediate adhesion to immortalized porcine bronchial epithelial cells (hTERT-PBECs). Pre-neutralizing M. hyopneumoniae with anti-rNOX antibody resulted in a more than 55% reduction in the adhesion rate of high- and low-virulence M. hyopneumoniae strains to hTERT-PBECs. Moreover, a significant difference appeared in the decline in CCU50 titer between virulent (168) and virulence-attenuated (168L) strains. NOX not only recognized and interacted with host fibronectin but also induced cellular oxidative stress and apoptosis in hTERT-PBECs. The release of lactate dehydrogenase by NOX in hTERT-PBECs was positively correlated with the virulence of M. hyopneumoniae strains. Conclusions NOX is considered to be a potential virulence factor of M. hyopneumoniae and may play a significant role in mediating its pathogenesis.https://doi.org/10.1186/s12917-022-03230-7Mycoplasma hyopneumoniaeNADH oxidaseAdhesionVirulence factor |
| spellingShingle | Fei Hao Xing Xie Zhixin Feng Rong Chen Yanna Wei Jin Liu Qiyan Xiong Guoqing Shao Johnson Lin NADH oxidase of Mycoplasma hyopneumoniae functions as a potential mediator of virulence BMC Veterinary Research Mycoplasma hyopneumoniae NADH oxidase Adhesion Virulence factor |
| title | NADH oxidase of Mycoplasma hyopneumoniae functions as a potential mediator of virulence |
| title_full | NADH oxidase of Mycoplasma hyopneumoniae functions as a potential mediator of virulence |
| title_fullStr | NADH oxidase of Mycoplasma hyopneumoniae functions as a potential mediator of virulence |
| title_full_unstemmed | NADH oxidase of Mycoplasma hyopneumoniae functions as a potential mediator of virulence |
| title_short | NADH oxidase of Mycoplasma hyopneumoniae functions as a potential mediator of virulence |
| title_sort | nadh oxidase of mycoplasma hyopneumoniae functions as a potential mediator of virulence |
| topic | Mycoplasma hyopneumoniae NADH oxidase Adhesion Virulence factor |
| url | https://doi.org/10.1186/s12917-022-03230-7 |
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