Systemic and local mucosal immune responses induced by orally delivered Bacillus subtilis spore expressing leucine aminopeptidase 2 of Clonorchis sinensis (original) (raw)

Abstract

Human clonorchiasis caused by Clonorchis sinensis (C. sinensis) has been increasingly prevalent in recent years so that an effective measure is essential and urgent to control the infectious disease. Oral delivery of antigens from C. sinensis may be an important approach to effectively induce both systemic and local immune responses to anti-infection of the parasite. In the current study, we used Bacillus subtilis (B. subtilis) spores as a delivery vehicle to introduce leucine aminopeptidase 2 of C. sinensis (_Cs_LAP2), an excretory/secretory antigen with high immunogenicity, expressing on their surface. SDS-PAGE, western blotting, and flow cytometry indicated that _Cs_LAP2 was successfully expressed on the surface of B. subtilis spores (CotC-_Cs_LAP2 spores). BALB/c mice were treated with spores intragastrically. On day 31 after the treatment, we found that mice intragastrically treated with CotC-_Cs_LAP2 spores exhibited higher IgG, IgG1, IgG2a, and IgA level in sera as well as higher sIgA level in bile and intestinal lavage fluid compared to mice orally administrated with spores not expressing _Cs_LAP2 (CotC spores) and naïve mice. The peak titer of IgG/IgA presented on day 31/49 after oral administration. IgG1 level was lower than IgG2a in group administrated with CotC-_Cs_LAP2 spores. sIgA-secreting cells were obviously observed in intestinal epithelium of mice orally treated with CotC-_Cs_LAP2 spores. After incubated with CotC-_Cs_LAP2, the levels of IFN-γ, IL-6, IL-10, IL-17A, and TNF significantly increased in the supernatant of splenocytes isolated from mice orally treated with CotC-_Cs_LAP2 spores, while there was no statistically significant difference of IL-4 level representing Th2 response among the groups. Our study demonstrated that oral administration of _Cs_LAP2 delivered by B. subtilis spore elicited obvious systemic and local mucosal immunity. Secretory IgA and Th1-Th17 cellular immunity might involved in mechanisms of the immune response.

Access this article

Log in via an institution

Subscribe and save

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

Download references

Acknowledgments

This work was supported by the National Key Basic Research and Development Project of China (973 project, No. 2010CB530000), National Natural Science Foundation of China (No.81101270 and No. 81171602), the National S & T Major Program (2012ZX10004-220), Fundamental Research Funds for the Central Universities of China (No. 3164015 and No. 3161036) and Innovative Research Teams Project of South Wisdom Valley, Shunde, Guangdong province (2013CXTD03).

Conflict of interests

The authors declare that we have no competing interests.

Author information

Authors and Affiliations

  1. Department of Parasitology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangdong, People’s Republic of China
    Hongling Qu, Yanquan Xu, Hengchang Sun, Jinsi Lin, Jinyun Yu, Zeli Tang, Chi Liang, Shan Li, Wenjun Chen, Xuerong Li, Zhongdao Wu, Yan Huang & Xinbing Yu
  2. Key Laboratory of Tropical Disease Control (Sun Yat-Sen University), Ministry of Education, Guangzhou, 510080, China
    Hongling Qu, Yanquan Xu, Hengchang Sun, Jinsi Lin, Jinyun Yu, Zeli Tang, Chi Liang, Shan Li, Wenjun Chen, Xuerong Li, Zhongdao Wu, Yan Huang & Xinbing Yu
  3. Department of Parasitology, Guangxi Medical University, Nanning City, Guangxi Province, 530021, China
    Jiqing Shen

Authors

  1. Hongling Qu
  2. Yanquan Xu
  3. Hengchang Sun
  4. Jinsi Lin
  5. Jinyun Yu
  6. Zeli Tang
  7. Jiqing Shen
  8. Chi Liang
  9. Shan Li
  10. Wenjun Chen
  11. Xuerong Li
  12. Zhongdao Wu
  13. Yan Huang
  14. Xinbing Yu

Corresponding author

Correspondence toXinbing Yu.

Additional information

Hongling Qu and Yanquan Xu contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Fig. S1

Construction strategy of recombinant plasmid of PEB03-CotC-_Cs_LAP2. Owning to the limited sites of restrict endonucleases in PEB03, the CotC-_Cs_LAP2 fusion gene was firstly inserted into multiple cloning site of pBluescript II SK (-). After digested with Sal I and Sac I, the CotC-_Cs_LAP2 fusion gene was sub-cloned into PEB03 shuttle vector. (DOC 401 kb)

Fig. S2

PCR amplification of CDS of CotC and identification of PEB03-CotC-_Cs_LAP2 by digestion. a: lane 1, DNA marker; lane 2, specific PCR production of CotC (397 bp). b: lane 1, DNA marker; lane 2, recombinant PEB03-CotC-_Cs_LAP2 plasmid; lane 3, PEB03-CotC-_Cs_LAP2 plasmid digested by Sal I and Sac I. The arrow indicated CotC-_Cs_LAP2 fusion gene (1967 bp). (DOC 359 kb)

Rights and permissions

About this article

Cite this article

Qu, H., Xu, Y., Sun, H. et al. Systemic and local mucosal immune responses induced by orally delivered Bacillus subtilis spore expressing leucine aminopeptidase 2 of Clonorchis sinensis .Parasitol Res 113, 3095–3103 (2014). https://doi.org/10.1007/s00436-014-3975-9

Download citation

Keywords