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Cloning, expression, and characterization of Schistosoma japonicum tegument protein phosphodiesterase-5

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Abstract

The tegument proteins of schistosomes are regarded as potential vaccine candidates and drug targets to control schistosomiasis. Nucleotide pyrophosphatase/phosphodiesterase-5 (NPP-5), which belongs to a multigene family of nucleotide pyrophosphatase/phosphodiesterases (NPPs), is important in the hydrolysis of pyrophosphate or phosphodiester bonds in nucleotides and their derivatives. In the present study, SjNPP-5, identified as one of the tegument proteins of Schistosoma japonicum in our previous proteomic studies, was cloned on a fragment of 1,371 bp and expressed as a recombinant protein of 69 kDa. Real-time RT-PCR analysis showed that SjNPP-5 was up-regulated at 21–42 days, and the expression level in 42-day-old male worms was almost nine times higher than that in females. Western blot analysis revealed that rSjNPP-5 had good antigenicity. Immunofluorescence analysis found that SjNPP-5 was a membrane-associated antigen mainly distributed on the surface of the male adult worm of S. japonicum. BALB/c mice vaccinated with rSjNPP-5 three times showed a 29.90% worm reduction (P < 0.05) and a 26.21% egg count reduction (P > 0.05). Immunization with rSjNPP-5 induced a mixed Th1/Th2 response in which Th1 was dominant. The response was characterized by a reduced IgG1/IgG2a ratio and elevated production of cytokines IFN-γ and IL-4. This study suggested that SjNPP-5 may be important in schistosome development, and further investigations are required to fully understand the function of this molecule.

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References

  • Basch PF (1990) Why do schistosomes have separate sexes? Parasitol Today 6:160–163

    Article  PubMed  CAS  Google Scholar 

  • Bergquist NR, Leonardo LR, Mitchell GF (2005) Vaccine-linked chemotherapy: can schistosomiasis control benefit from an integrated approach? Trends Parasitol 21:112–117

    Article  PubMed  CAS  Google Scholar 

  • Braschi S, Borges WC, Wilson RA (2006) Proteomic analysis of the schistosome tegument and its surface membranes. Mem Inst Oswaldo Cruz 101(Suppl 1):205–212

    Article  PubMed  CAS  Google Scholar 

  • Cardoso FC, Macedo GC, Gava E, Kitten GT, Mati VL, de Melo AL, Caliari MV, Almeida GT, Venancio TM, Verjovski-Almeida S, Oliveira SC (2008) Schistosoma mansoni tegument protein Sm29 is able to induce a Th1-type of immune response and protection against parasite infection. PLoS Negl Trop Dis 2:e308

    Article  PubMed  Google Scholar 

  • Coffman RL, Carty J (1986) A T cell activity that enhances polyclonal IgE production and its inhibition by interferon-gamma. J Immunol 136:949–954

    PubMed  CAS  Google Scholar 

  • Duan DM, Xiong LS, Wu J (2007) Advances of the ecto-nucleotide pyrophosphatases/phosphodiesterases. Letters in Biotechnology 18:277–280

    CAS  Google Scholar 

  • Finkelman FD, Holmes J, Katona IM, Urban JF Jr, Beckmann MP, Park LS, Schooley KA, Coffman RL, Mosmann TR, Paul WE (1990) Lymphokine control of in vivo immunoglobulin isotype selection. Annu Rev Immunol 8:303–333

    Article  PubMed  CAS  Google Scholar 

  • Goding JW, Grobben B, Slegers H (2003) Physiological and pathophysiological functions of the ecto-nucleotide pyrophosphatase/phosphodiesterase family. Biochim Biophys Acta 1638:1–19

    PubMed  CAS  Google Scholar 

  • Golding B, Scott DE (1995) Vaccine strategies: targeting helper T cell responses. Ann N Y Acad Sci 754:126–137

    Article  PubMed  CAS  Google Scholar 

  • He YX, Yang HZ (1980) Physiological studies on the post-cercarial development of Schistosoma japonicum. Acta Zoologica Sinica 26:32–39

    Google Scholar 

  • Hockley DJ, McLaren DJ (1973) Schistosoma mansoni: changes in the outer membrane of the tegument during development from cercaria to adult worm. Int J Parasitol 3:13–25

    Article  PubMed  CAS  Google Scholar 

  • Jankovic D, Kullberg MC, Noben-Trauth N, Caspar P, Ward JM, Cheever AW, Paul WE, Sher A (1999) Schistosome-infected IL-4 receptor knockout (KO) mice, in contrast to IL-4 KO mice, fail to develop granulomatous pathology while maintaining the same lymphokine expression profile. J Immunol 163:337–342

    PubMed  CAS  Google Scholar 

  • Jones MK, Gobert GN, Zhang L, Sunderland P, McManus DP (2004) The cytoskeleton and motor proteins of human schistosomes and their roles in surface maintenance and host-parasite interactions. Bioessays 26:752–765

    Article  PubMed  CAS  Google Scholar 

  • Kumar S, Tamura K, Nei M (2004) MEGA3: Integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment. Brief Bioinform 5:150–163

    Article  PubMed  CAS  Google Scholar 

  • Lopes DO, Paiva LF, Martins MA, Cardoso FC, Rajao MA, Pinho JM, Caliari MV, Correa-Oliveira R, Mello SM, Leite LC, Oliveira SC (2009) Sm21.6 a novel EF-hand family protein member located on the surface of Schistosoma mansoni adult worm that failed to induce protection against challenge infection but reduced liver pathology. Vaccine 27:4127–4135

    Article  PubMed  CAS  Google Scholar 

  • Loukas A, Tran M, Pearson MS (2007) Schistosome membrane proteins as vaccines. Int J Parasitol 37:257–263

    Article  PubMed  CAS  Google Scholar 

  • Ohe Y, Ohnishi H, Okazawa H, Tomizawa K, Kobayashi H, Okawa K, Matozaki T (2003) Characterization of nucleotide pyrophosphatase-5 as an oligomannosidic glycoprotein in rat brain. Biochem Biophys Res Commun 308:719–725

    Article  PubMed  CAS  Google Scholar 

  • Qian MB, Hu W (2008) The structure and function of schistosome tegument and related proteomic study. Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi 26:466–471

    PubMed  Google Scholar 

  • Rofatto HK, Tararam CA, Borges WC, Wilson RA, Leite LC, Farias LP (2009) Characterization of phosphodiesterase-5 as a surface protein in the tegument of Schistosoma mansoni. Mol Biochem Parasitol 166:32–41

    Article  PubMed  CAS  Google Scholar 

  • Sakagami H, Aoki J, Natori Y, Nishikawa K, Kakehi Y, Arai H (2005) Biochemical and molecular characterization of a novel choline-specific glycerophosphodiester phosphodiesterase belonging to the nucleotide pyrophosphatase/phosphodiesterase family. J Biol Chem 280:23084–23093

    Article  PubMed  CAS  Google Scholar 

  • Sher A, Coffman RL, Hieny S, Scott P, Cheever AW (1990) Interleukin 5 is required for the blood and tissue eosinophilia but not granuloma formation induced by infection with Schistosoma mansoni. Proc Natl Acad Sci USA 87:61–65

    Article  PubMed  CAS  Google Scholar 

  • Smythies LE, Coulson PS, Wilson RA (1992) Monoclonal antibody to IFN-gamma modifies pulmonary inflammatory responses and abrogates immunity to Schistosoma mansoni in mice vaccinated with attenuated cercariae. J Immunol 149:3654–3658

    PubMed  CAS  Google Scholar 

  • Stefan C, Jansen S, Bollen M (2005) NPP-type ectophosphodiesterases: unity in diversity. Trends Biochem Sci 30:542–550

    Article  PubMed  CAS  Google Scholar 

  • Tran MH, Pearson MS, Bethony JM, Smyth DJ, Jones MK, Duke M, Don TA, McManus DP, Correa-Oliveira R, Loukas A (2006) Tetraspanins on the surface of Schistosoma mansoni are protective antigens against schistosomiasis. Nat Med 12:835–840

    Article  PubMed  CAS  Google Scholar 

  • Umezu-Goto M, Kishi Y, Taira A, Hama K, Dohmae N, Takio K, Yamori T, Mills GB, Inoue K, Aoki J, Arai H (2002) Autotaxin has lysophospholipase D activity leading to tumor cell growth and motility by lysophosphatidic acid production. J Cell Biol 158:227–233

    Article  PubMed  CAS  Google Scholar 

  • van der Werf MJ, de Vlas SJ, Brooker S, Looman CW, Nagelkerke NJ, Habbema JD, Engels D (2003) Quantification of clinical morbidity associated with schistosome infection in sub-Saharan Africa. Acta Trop 86:125–139

    Article  PubMed  Google Scholar 

  • Wang L, Utzinger J, Zhou XN (2008) Schistosomiasis control: experiences and lessons from China. Lancet 372:1793–1795

    Article  PubMed  Google Scholar 

  • Wang X, Huang C, Sun B, Gu Y, Cui Y, Zhao X, Li Y, Zhang S (2010) The effect of high gravidity on the carcinogenesis of mammary gland in TA2 mice. Am J Reprod Immunol 63:396–409

    Article  PubMed  CAS  Google Scholar 

  • WHO (2002) TDR strategic direction for research: schistosomiasis. World Health Organization, Geneve

    Google Scholar 

  • Wilson RA, Barnes PE (1974) An in vitro investigation of dynamic processes occurring in the schistosome tegument, using compounds known to disrupt secretory processes. Parasitology 68:259–270

    PubMed  CAS  Google Scholar 

  • Xu X, Zhang D, Sun W, Zhang Q, Zhang J, Xue X, Shen L, Pan W (2009) A Schistosoma japonicum chimeric protein with a novel adjuvant induced a polarized Th1 immune response and protection against liver egg burdens. BMC Infect Dis 9:54

    Article  PubMed  Google Scholar 

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Acknowledgments

We thank Dr. Jinbiao Peng for valuable suggestions on the manuscript and Yaojun Shi from the Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, for technical assistance with parasite collection. This work was supported by the National Basic Research Program of China (no. 2007CB513108), Special Fund for Agro-Scientific Research in the Public Interest (no. 200903036), and National Natural Science Foundation of China (no. 30671581).

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Authors and Affiliations

  1. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of Agriculture, 518 Ziyue road, Minhang, Shanghai, 200241, People’s Republic of China

    Min Zhang, Yanhui Han, Zhu Zhu, Dong Li, Yang Hong, Xiujuan Wu, Zhiqiang Fu & Jiaojiao Lin

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  1. Min Zhang
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  2. Yanhui Han
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  3. Zhu Zhu
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  5. Yang Hong
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  8. Jiaojiao Lin
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Correspondence to Jiaojiao Lin.

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Zhang, M., Han, Y., Zhu, Z. et al. Cloning, expression, and characterization of Schistosoma japonicum tegument protein phosphodiesterase-5. Parasitol Res 110, 775–786 (2012). https://doi.org/10.1007/s00436-011-2552-8

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