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Expression, immunolocalization, and serological reactivity of a novel sphingomyelin phosphodiesterase-like protein, an excretory/secretory antigen from Clonorchis sinensis

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Abstract

Clonorchiasis, caused by Clonorchis sinensis infection, is a zoonotic parasitic disease of hepatobiliary system in which the proteins released by adult are major pathogenetic factors. In this study, we first characterized a putative sphingomyelin phosphodiesterase (CsSMPase) A-like secretory protein, which was highly expressed in the adult worm. The full-length gene was cloned. The putative protein is of relatively low homology comparing with SMPase from other species, and of rich T cell and B cell epitopes, suggesting that it is an antigen of strong antigenicity. The complete coding sequence of the gene was expressed in the Escherichia coli. The recombinant CsSMPase (rCsSMPase) can be recognized by C. sinensis-infected serum, and the protein immunoserum can recognize a specific band in excretory/secretory products (ESPs) of C. sinensis adult by western blotting. Immunolocalization revealed that CsSMPase was not only localized on tegument, ventral sucker of metacercaria, and the intestine of adult but also on the nearby epithelium of bile duct of the infected Sprague–Dawley rats, implying that CsSMPase was mainly secreted and excreted through adult intestine and directly interacted with bile duct epithelium. Although immunized rats evoked high level antibody response, the antigen level was low in clonorchiasis patients. And the sensitivity and specificity of rCsSMPase were 50.0 % (12/24) and 88.4 % (61/69), in sera IgG-ELISA, respectively. It is likely due to the fact that CsSMPase binding to the plasma membrane of biliary epithelium decreases the antigen immune stimulation.

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Acknowledgments

This work was supported by China National Great Basic Research Program (973 program, no. 2010CB530003)

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Authors and Affiliations

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

    Yanwei Huang, Yuzhe Li, Mei Yang, Ting Li, Suxiang Zeng, Xinbing Yu & Xuchu Hu

  2. Key Laboratory of Tropical Disease Control (Sun Yat-Sen University), Ministry of Education, Guangzhou, Guangdong, China

    Yanwei Huang, Yuzhe Li, Mei Yang, Ting Li, Suxiang Zeng, Xinbing Yu & Xuchu Hu

  3. Department of Pathology and Laboratory Medicine, Guangdong Provincial People’s Hospital, Guangzhou, China

    Youwei Zheng

  4. Department of Dermatology and Venereology, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China

    Huaiqiu Huang

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  1. Yanwei Huang
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Correspondence to Huaiqiu Huang or Xuchu Hu.

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Huang, Y., Zheng, Y., Li, Y. et al. Expression, immunolocalization, and serological reactivity of a novel sphingomyelin phosphodiesterase-like protein, an excretory/secretory antigen from Clonorchis sinensis . Parasitol Res 112, 2197–2206 (2013). https://doi.org/10.1007/s00436-013-3388-1

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  • DOI: https://doi.org/10.1007/s00436-013-3388-1

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