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Acta Parasitologica

, Volume 63, Issue 2, pp 252–260 | Cite as

Molecular cloning of enolase from Trichinella spiralis and the protective immunity in mice

  • Xuliang Zhang
  • Lixin Xu
  • Xiaokai Song
  • Xiangrui Li
  • Ruofeng YanEmail author
Article

Abstract

Trichinella spiralis, the main pathogen of trichinosis, infects a wide range of mammalian hosts and is one of the most widespread parasites worldwide. For parasites, glycolysis is the most important way to generate energy. Previous studies showed that some enzymes involved in the glycolytic pathway play roles in regulation the host immunity. In this paper, enolase from T. spiralis was cloned and the protective potentials were studied. One hundred and sixty ICR mice were divided into four groups and vaccinated with recombinant enolase (pET-ENO), eukaryotic recombinant plasmid encoding enolase (pVAX1- ENO) and negative controls (pVAX1 and PBS), respectively. Two weeks after the second immunization, each mouse was challenged orally with 200 muscle larvae (MLs) of T. spiralis. Results showed that mice vaccinated with pET-ENO and pVAX1-ENO induced specific antibodies of IgG, IgA, IgM, but no IgE. Subclasses of IgG antibodies showed that mice immunized with recombinant protein and recombinant plasmids induced a Th1/Th2 immune response. Concentrations of serum cytokines were detected and showed significant increase of IFN-γ, IL-4 and TGFβ1, while IL-17 in each group was not significantly different. Flow cytometric analysis showed significant increase of CD4+ and CD8+ T lymphocytes in the groups immunized with recombinant protein and recombinant plasmids. Challenge infection demonstrated that immunized groups had a reduced number of worm burdens. The reductions of larvae per gram muscle (LPG) in pET-ENO and pVAX1-ENO group were 17.7% and 15.8% when compared with PBS control.

Keywords

Trichinella spiralis Enolase DNA vaccination immune response 

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References

  1. Bien, J., Cabaj, W., Moskwa B. 2015. Proteomic analysis of potential immunoreactive proteins from muscle larvae and adult worms of Trichinella spiralis in experimentally infected pigs. Folia Parasitologica, 62, 022CrossRefGoogle Scholar
  2. Chen, N., Yuan, Z.G., Xu, M.J., Zhou, D.H., Zhang, X.X., Zhang, Y.Z., et al. 2012. Ascaris suum enolase is a potential vaccine candidate against ascariasis. Vaccine, 30, 3478–82CrossRefGoogle Scholar
  3. Choi, K.D., Lillehoj, H.S., Zalenga, D.S., 1999. Changes in local IFN-γ and TGF-β4 mRNA expression and intraepithelial lymphocytes following Eimeria acervulina infection. Veterinary Immunology and Immunopathology, 71, 263–275CrossRefGoogle Scholar
  4. Dea-Ayuela, M.A., Bolas-Fernandez F. 2005. Two-dimensional electrophoresis and mass spectrometry for the identification of species-specific Trichinella antigens. Veterinary Parasitology, 132, 43–49CrossRefGoogle Scholar
  5. Dennis, D.T., Despommier, D.D., Davis N. 1970. Infectivity of the newborn larva of Trichinella spiralis in the rat. Journal of Parasitology, 56, 974–977CrossRefGoogle Scholar
  6. Dupouy-Camet J. 2009. Presidential address of ICT12 Conference: “Trichinella and trichinellosis–A never ending story”. Veterinary Parasitology, 159, 194–196CrossRefGoogle Scholar
  7. Ferguson, D.J., Parmley, S.F., Tomavo S. 2002. Evidence for nuclear localisation of two stage-specific isoenzymes of enolase in Toxoplasma gondii correlates with active parasite replication. International Journal for Parasitology, 32, 1399–1410CrossRefGoogle Scholar
  8. Han, K., Xu, L., Yan, R., Song, X., Li X. 2012. Vaccination of goats with glyceraldehyde-3- phosphate dehydrogenase DNA vaccine induced partial protection against Haemonchus contortus. Veterinary Immunology and Immunopathology, 149, 177–185CrossRefGoogle Scholar
  9. Kalyanasundaram. A., Jawahar, S., Ilangopathy, M., Palavesam, A., Raman M. 2015. Comparative immunoprophylactic efficacy of Haemonchus contortus recombinant enolase (rHcENO) and Con A purified native glycoproteins in sheep. Experimental Parasitololgy, 154, 98–107CrossRefGoogle Scholar
  10. Mahana, N., Abd-Allah, H.A., Salah, M., Tallima, H., EIRidi R. 2016. Fasciola gigantica enolase is a major component of worm tegumental fraction protective against sheep fasciolosis. Acta Tropica, 58, 189–96CrossRefGoogle Scholar
  11. Moskwa, B., Gozdzik, K., Bien, J., Borecka, A., Gawor, J., Cabaj W. 2013. First report of Trichinella pseudospiralis in Poland, in red foxes (Vulpes vulpes). Acta Parasitologica, 58, 149–154CrossRefGoogle Scholar
  12. Muleke, C.I., Ruofeng, Y., Yanming, S., Guangwei, Z., Lixin, X., Xiangrui L., 2007. Vaccination of goats against Haemonchus contortus with a recombinant cysteine protease. Small Ruminant Research, 73, 95–102CrossRefGoogle Scholar
  13. Nakada, T., Na, I., Wu, Z., Takahashi, Y., 2005. Molecular cloning and functional expression of enolase from Trichinella spiralis. Parasitology Research, 96, 354–360CrossRefGoogle Scholar
  14. Pozio E. 2015. Trichinella spp. imported with live animals and meat. Veterinary Parasitology, 213, 46–55CrossRefGoogle Scholar
  15. Pozio, E., Darwin M.K. 2006. Systematics and Epidemiology of Trichinella. Advances in Parasitology, 63, 367–439CrossRefGoogle Scholar
  16. Pozio, E., Zarlenga D.S. 2005. Recent advances on the taxonomy, systematics and epidemiology of Trichinella. International Journal for Parasitology, 35, 1191–1204CrossRefGoogle Scholar
  17. Saad, A.E., Ashour, D.S., Abou Rayia, D.M., Bedeer A.E. 2016. Carbonic anhydrase enzyme as a potential therapeutic target for experimental trichinellosis. Parasitology Research, 115, 2331–2339CrossRefGoogle Scholar
  18. Santrac, V., Nedic, D.N., Maric, J., Nikolic, S., Stevanovic, O., Vasilev S., et al. 2015. The first report of Trichinella pseudospiralis presence in domestic swine and T. britovi in wild boar in Bosnia and Herzegovina. Acta Parasitologica, 60, 471–475CrossRefGoogle Scholar
  19. Sun, W., Song, X., Yan, R., Xu, L., Li X. 2011. Vaccination of goats with a glutathione peroxidase DNA vaccine induced partial protection against Haemonchus contortus infection. Veterinary Parasitology, 182, 239–247CrossRefGoogle Scholar
  20. Sun, Y., Yan, R., Charles, I.M., Zhao, G., Xu, L., Li X. 2007. Vaccination of goats with recombinant galectin antigen induces partial protection against Haemonchus contortus infection. Parasite Immunology, 29, 319–326CrossRefGoogle Scholar
  21. Tang, F., Xu, L., Yan, R., Song, X., Li X. 2013. A DNA vaccine co-expressing Trichinella spiralis MIF and MCD-1 with murine ubiquitin induces partial protective immunity in mice. Journal of Helminthology, 87, 24–33CrossRefGoogle Scholar
  22. Wang, L., Wang, X., Bi, K., Sun, X., Yang, J., Gu, Y., et al. 2016. Oral vaccination with attenuated Salmonella typhimurium-Delivered TsPmy DNA Vaccine elicits protective immunity against Trichinella spiralis in BALB/c Mice. PLoS Neglected Tropical Disease 10, e0004952CrossRefGoogle Scholar
  23. Wang, X., Chen, W., Tian, Y., Huang, Y., Li, X., Yu X. 2014. RNAi-mediated silencing of enolase confirms its biological importance in Clonorchis sinensis. Parasitology Research, 113, 1451–1458CrossRefGoogle Scholar
  24. Whitton, J.L., Rodriguez, F., Zhang, J., Hassett D.E. 1999. DNA immunization: mechanistic studies. Vaccine, 17, 1612–1619CrossRefGoogle Scholar
  25. Yang, J., Gu, Y., Yang, Y., Wei, J., Wang, S., Cui, S., et al. 2010a. Trichinella spiralis: immune response and protective immunity elicited by recombinant paramyosin formulated with different adjuvants. Experimental Parasitology, 124, 403–408CrossRefGoogle Scholar
  26. Yang, Y., Zhang, Z., Yang, J., Chen, X., Cui, S., Zhu X. 2010b. Oral vaccination with Ts87 DNA vaccine delivered by attenuated Salmonella typhimurium elicits a protective immune response against Trichinella spiralis larval challenge. Vaccine, 28, 2735–2742CrossRefGoogle Scholar
  27. Zhang, C., Gu, Y., Tang, J., Lu, F., Cao, Y., Zhou, H., et al. 2106. Production of Plasmodium vivax enolase in Escherichia coli and its protective properties. Human Vaccines & Immunotherapeutics, 12, 2855–2861CrossRefGoogle Scholar
  28. Zhao, G., Yan, R., Muleke, C.I., Sun, Y., Xu, L., Li X. 2012. Vaccination of goats with DNA vaccines encoding H11 and IL-2 induces partial protection against Haemonchus contortus infection. Veterinary Journal, 191, 94–100CrossRefGoogle Scholar
  29. Zhu, X., Long, G., Xu, F., Gui Y. 1993. Trichinellosis. 1st edn. Zhengzhou, Henan Science and Technology Press, pp. 258–260Google Scholar

Copyright information

© Witold Stefański Institute of Parasitology, Polish Academy of Sciences 2018

Authors and Affiliations

  • Xuliang Zhang
    • 1
  • Lixin Xu
    • 1
  • Xiaokai Song
    • 1
  • Xiangrui Li
    • 1
  • Ruofeng Yan
    • 1
    Email author
  1. 1.MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary MedicineNanjing Agricultural UniversityNanjingPeople’s Republic of China

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