Abstract
Mycoplasma hyorhinis is commonly found in the respiratory tract of pigs and is the etiological agent of polyserositis. The metabolic enzymes of M. hyorhinis may play important roles in host-pathogen interactions. We immunized BALB/c mice with sodium deoxycholate-extracted antigens (DOC-Ags) and screened 10 hybridomas that secreted antibodies against various M. hyorhinis proteins. Pyruvate dehydrogenase E1 complex subunit alpha (PDHA) was identified as the protein that reacted with five of the 10 monoclonal antibodies (mAbs). Sequence analysis indicated that PDHA was highly conserved among M. hyorhinis strains, but not among other mycoplasmas. We predicted the three-dimensional structure of PDHA and identified three epitopes (277RTEEEEK283, 299KDKKYITDE307, and 350LKEQKQHAKDY360). The mAb 1H12 we generated was used to detect M. hyorhinis PDHA in vitro and in piglets infected with M. hyorhinis. We observed that PDHA was mainly located in the epithelial cells of the lungs. Our results indicate that the mAbs we generated could be used to further investigate the structure and function of M. hyorhinis PDHA. In addition, they could be used in the differential diagnosis of M. hyorhinis and other mycoplasmas.
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Acknowledgments
This work was supported by the special funds of technical innovative talents of Harbin (grant no.: 2014RFQYJ083) and the special fund for basic scientific research business of central public research institutes (grant no.: 1610302015014).
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Animal experiments used in the present study were approved by the Animal Ethics Committee of Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences (Heilongjiang-SYXK-2006-032) and were performed in accordance with relevant animal ethics guidelines.
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Dongjie Chen and Yanwu Wei contributed equally to this work.
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Chen, D., Wei, Y., Huang, L. et al. Characterization and application of monoclonal antibodies against Mycoplasma hyorhinis pyruvate dehydrogenase E1 complex subunit alpha. Appl Microbiol Biotechnol 100, 3587–3597 (2016). https://doi.org/10.1007/s00253-015-7263-0
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DOI: https://doi.org/10.1007/s00253-015-7263-0