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Biochemical and immunological characterization of annexin B30 from Clonorchis sinensis excretory/secretory products

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Abstract

Clonorchis sinensis has been classified as group I biological carcinogen for cholangiocarcinoma by the World Health Organization. Biological studies on excretory/secretory products (ESPs) enabled us to understand the pathogenesis mechanism of C. sinensis and develop new strategies for the prevention of clonorchiasis. In this study, sequence analysis showed that annexin B30 from C. sinensis (CsANXB30) is composed of four annexin repeats which were characterized by type II and III Ca2+-binding sites or KGD motif with the capability of Ca2+-binding. In addition, immunoblot assay revealed that recombinant CsANXB30 (rCsANXB30) could be recognized by the sera from rats infected with C. sinensis and the sera from rats immunized by CsESPs. Real-time PCR showed that its transcriptional level was the highest at the stage of metacercaria. Immunofluorescence assay was employed to confirm that CsANXB30 was distributed in the tegument, intestine, and egg of adult worms, as well as the tegument and vitellarium of metacercaria. rCsANXB30 was able to bind phospholipid in a Ca2+-dependent manner and human plasminogen in a dose-dependent manner. Moreover, cytokine and antibody measurements indicated that rats subcutaneously immunized with rCsANXB30 developed a strong IL-10 production in spleen cells and a high level of IgG1 isotype, indicating that rCsANXB30 could trigger specific humoral and cellular immune response in rats. The present results implied that CsANXB30 might be involved in a host–parasite interaction and affected the immune response of the host during C. sinensis infection.

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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 Guangdong Natural Science Foundation of China (No. S2012010008504).

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The authors declare that we have no competing interests.

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  1. Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, 510080, China

    Lei He, Mengyu Ren, Xueqing Chen, Xiaoyun Wang, Shan Li, Jinsi Lin, Chi Liang, Meng Bian, Yan Huang, Zhongdao Wu, Xuerong Li & Xinbing Yu

  2. Key Laboratory of Tropical Disease Control, Ministry of Education, Sun Yat-sen University, Guangzhou, 510080, China

    Lei He, Mengyu Ren, Xueqing Chen, Xiaoyun Wang, Shan Li, Jinsi Lin, Chi Liang, Meng Bian, Yan Huang, Zhongdao Wu, Xuerong Li & Xinbing Yu

  3. Department of Pathogen Biology, Hainan Medical College, Haikou, Hainan Province, 571199, China

    Pei Liang

  4. Department of Clinical Laboratory, the Second People’s Hospital of Hefei, Hefei, Anhui Province, 230011, China

    Yue Hu

  5. Department of Infectious Diseases, the Affiliated Guangzhou No.8 People’s Hospital, Guangzhou Medical University, Guangzhou, 510060, China

    Huali Lei

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  1. Lei He
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Correspondence to Xuerong Li or Xinbing Yu.

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Lei He and Mengyu Ren contributed equally to this article.

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He, L., Ren, M., Chen, X. et al. Biochemical and immunological characterization of annexin B30 from Clonorchis sinensis excretory/secretory products. Parasitol Res 113, 2743–2755 (2014). https://doi.org/10.1007/s00436-014-3935-4

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