Parasitology Research

, Volume 113, Issue 5, pp 1749–1757

Identification, immunolocalization, and immunological characterization of nitric oxide synthase-interacting protein from Clonorchis sinensis

  • Meng Bian
  • Shan Li
  • Xiaoyun Wang
  • Yanquan Xu
  • Wenjun Chen
  • Chenhui Zhou
  • Xueqing Chen
  • Lei He
  • Jin Xu
  • Chi Liang
  • Zhongdao Wu
  • Yan Huang
  • Xuerong Li
  • Xinbing Yu
Original Paper

DOI: 10.1007/s00436-014-3820-1

Cite this article as:
Bian, M., Li, S., Wang, X. et al. Parasitol Res (2014) 113: 1749. doi:10.1007/s00436-014-3820-1

Abstract

Recently, accumulating evidences indicate that nitric oxide (NO) is a potent mediator with diverse roles in regulating cellular functions, signaling pathways, and variety of pathological processes. In the present study, using data from the published genomic for Clonorchis sinensis (C. sinensis), we investigated a gene encoding nitric oxide synthase-interacting protein (NOSIP) of C. sinensis. Recombinant CsNOSIP (rCsNOSIP) was expressed and purified from Escherichia coli BL21. The open reading frame of CsNOSIP comprises 867 bp which encodes 289 amino acids and shares 72.9, 45.2, 47, 46.4, and 45.8 % identity with NOSIP from Schistosoma mansoni, Xenopus laevis, Rattus norvegicus, Mus musculus, and Homo sapiens, respectively. Bioinformatics analysis suggested that the full-length sequence contains an eNOS-interacting domain and numerous B-cell epitopes. Quantitative RT-PCR indicated that CsNOSIP differentially transcribed throughout the adult worms, metacercariae, and egg stages of C. sinensis, and were highly expressed in the adult worms. Moreover, western blot analysis showed that the rCsNOSIP could be detected by the serum from BALB/c mice infected with C. sinensis and the serum from BALB/c mice immunized with excretory/secretory products (ESPs). Furthermore, immunolocalization assay showed that CsNOSIP was specifically localized in the intestine, vitellarium, and eggs of adult worm. Both immunoblot and immunolocalization results demonstrated that CsNOSIP was one component of ESPs of C. sinensis, which could be supported by SignalP analysis. Moreover, analysis of the antibody subclass and cytokine profile demonstrated that subcutaneously immunized BALB/c mice with rCsNOSIP could significantly enhance serum IgG1 level and up-regulate expression of IL-4 and IL-6 in the splenocytes. Our results suggested that CsNOSIP was an important antigen exposed to host immune system and probably involved in immune regulation of host by inducing Th2-polarized immune response.

Keywords

Clonorchis sinensis Nitric oxide-interacting protein Immune response Characterization 

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Meng Bian
    • 1
    • 2
  • Shan Li
    • 1
    • 2
  • Xiaoyun Wang
    • 1
    • 2
  • Yanquan Xu
    • 1
    • 2
  • Wenjun Chen
    • 1
    • 2
  • Chenhui Zhou
    • 1
    • 2
    • 3
  • Xueqing Chen
    • 1
    • 2
  • Lei He
    • 1
    • 2
  • Jin Xu
    • 1
    • 2
  • Chi Liang
    • 1
    • 2
  • Zhongdao Wu
    • 1
    • 2
  • Yan Huang
    • 1
    • 2
  • Xuerong Li
    • 1
    • 2
  • Xinbing Yu
    • 1
    • 2
  1. 1.Department of Zhongshan School of MedicineSun Yat-sen UniversityGuangzhouPeople’s Republic of China
  2. 2.Key Laboratory for Tropical Diseases Control, Ministry of EducationSun Yat-sen UniversityGuangzhouPeople’s Republic of China
  3. 3.School of NursingGuangdong Medical CollegeDongguanPeople’s Republic of China

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