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Oral delivery of Bacillus subtilis spore expressing enolase of Clonorchis sinensis in rat model: induce systemic and local mucosal immune responses and has no side effect on liver function

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Abstract

Caused by the consumption of raw or undercooked freshwater fish containing infective metacercariae of Clonorchis sinensis, human clonorchiasis remains a major public health problem in China. In previous study, we had expressed enolase from C. sinensis (CsENO) on the surface of Bacillus subtilis spore and the recombinant spore induced a pronounced protection in terms of reduced worm burden and eggs per gram feces, suggesting B. subtilis spore as an ideal vehicle for antigen delivery by oral treatment and CsENO as a promising vaccine candidate against clonorchiasis. In the current study, we detected CsENO-specific IgG and IgA levels both in serum and in intestinal mucus from rats orally administrated with B. subtilis spore surface expressing CsENO by ELISA. Lysozyme levels in serum and in intestinal mucus were analyzed too. In addition, IgA-secreting cells in intestine epithelium of the rats were detected by immunohistochemistry assay. The intestinal villi lengths of duodenum, jejunum, and ileum were also measured. Rats orally treated with B. subtilis spore or normal saline were used as controls. Our results showed that, compared with the control groups, oral administration of B. subtilis spore expressing CsENO induced both systemic and local mucosal immune response. The recombinant spores also enhanced non-specific immune response in rats. The spores had no side effect on liver function. Moreover, it might facilitate food utilization and digestion of the rats. Our work will pave the way to clarify the involved mechanisms of protective efficacy elicited by B. subtilis spore expressing CsENO and encourage us to carry out more assessment trails of the oral treated spore to develop vaccine against clonorchiasis.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 81101270 and No. 81171602), Science and Technology Planning Project of Guangdong Province (No.2013B010404010), Fundamental Research Funds for the Central Universities of China (No. 3161036), Innovative Research Teams Project of South Wisdom Valley, Shunde, Guangdong province (2013CXTD03), National Key Basic Research and Development Project of China (973 project, No. 2010CB530000), and the National S & T Major Program (2012ZX10004-220).

Conflict of interest

The authors declare that they have no competing interests.

Author information

Correspondence to Xinbing Yu.

Additional information

Jinyun Yu and Tingjin Chen contributed equally to this work.

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Yu, J., Chen, T., Xie, Z. et al. Oral delivery of Bacillus subtilis spore expressing enolase of Clonorchis sinensis in rat model: induce systemic and local mucosal immune responses and has no side effect on liver function. Parasitol Res 114, 2499–2505 (2015). https://doi.org/10.1007/s00436-015-4449-4

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Keywords

  • Clonorchis sinensis
  • Bacillus subtilis spore
  • Enolase
  • Oral immunization
  • Immune response
  • Liver function