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Comparative analysis of immune effects in mice model: Clonorchis sinensis cysteine protease generated from recombinant Escherichia coli and Bacillus subtilis spores

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Abstract

Clonorchiasis remains a nonnegligible public health problem in endemic areas. Cysteine protease of Clonorchis sinensis (CsCP) plays indispensable roles in the parasitic physiology and pathology, and has been exploited as a promising drug and vaccine candidate. In recent years, development of spore-based vaccines against multiple pathogens has attracted many investigators’ interest. In previous studies, the recombinant Escherichia coli (BL21) and Bacillus subtilis spores expressing CsCP have been successfully constructed, respectively. In this study, the immune effects of CsCP protein purified from recombinant BL21 (rCsCP) and B. subtilis spores presenting CsCP (B.s-CsCP) in Balb/c mice model were conducted with comparative analysis. Levels of specific IgG, IgG1 and IgG2a were significantly increased in sera from both rCsCP and B.s-CsCP intraperitoneally immunized mice. Additionally, recombinant spores expressing abundant fusion CsCP (0.03125 pg/spore) could strongly enhance the immunogenicity of CsCP with significantly higher levels of IgG and isotypes. Compared with rCsCP alone, intraperitoneal administration of mice with spores expressing CsCP achieved a better effect of fighting against C. sinensis infection by slowing down the process of fibrosis. Our results demonstrated that a combination of Th1/Th2 immune responses could be elicited by rCsCP, while spores displaying CsCP prominently induced Th1-biased specific immune responses, and the complex cytokine network maybe mediates protective immune responses against C. sinensis. This work further confirmed that the usage of B. subtilis spores displaying CsCP is an effective way to against C. sinensis.

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Acknowledgments

This work was supported by the National Key Basic Research and Development Project of China (973 project; No.2010CB530000), the Science and Technology Planning Project of Guangdong Province (No. 2013B010404010, No. 2014B020203001 and No. 2016A050502008) and Innovative Research Teams Project of South Wisdom Valley, Shunde, Guangdong province (2013CXTD03).

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  1. Zhanshuai Wu and Zeli Tang contributed equally to this work.

Authors and Affiliations

  1. School of Life Sciences, Sun Yat-Sen University, Guangzhou, Guangdong, China

    Zhanshuai Wu

  2. Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China

    Zeli Tang, Mei Shang, Lu Zhao, Lina Zhou, Xiangzhan Kong, Zhipeng Lin, Hengchang Sun, Tingjin Chen, Jin Xu, Xuerong Li, Yan Huang & Xinbing Yu

  3. Key Laboratory for Tropical Diseases Control, Sun Yat-sen University, Ministry of Education, Guangzhou, Guangdong, China

    Zeli Tang, Mei Shang, Lu Zhao, Lina Zhou, Xiangzhan Kong, Zhipeng Lin, Hengchang Sun, Tingjin Chen, Jin Xu, Xuerong Li, Yan Huang & Xinbing Yu

  4. Provincial Engineering Technology Research Center for Biological Vector Control, Guangzhou, Guangdong, 510080, China

    Zeli Tang, Mei Shang, Lu Zhao, Lina Zhou, Xiangzhan Kong, Zhipeng Lin, Hengchang Sun, Tingjin Chen, Jin Xu, Xuerong Li, Yan Huang & Xinbing Yu

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  1. Zhanshuai Wu
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Corresponding authors

Correspondence to Yan Huang or Xinbing Yu.

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All animal experiments were conducted under the Institutional Animal Care and Use Committee of Sun Yat-Sen University (Permit Number: SYXK (Guangdong) 2010–0107).

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The authors declare that they have no competing interests.

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Wu, Z., Tang, Z., Shang, M. et al. Comparative analysis of immune effects in mice model: Clonorchis sinensis cysteine protease generated from recombinant Escherichia coli and Bacillus subtilis spores. Parasitol Res 116, 1811–1822 (2017). https://doi.org/10.1007/s00436-017-5445-7

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