Advertisement

Parasitology Research

, Volume 114, Issue 5, pp 2007–2013 | Cite as

Biochemical and functional characterization of the glutathione S-transferase from Trichinella spiralis

  • Jing Cui
  • Ling Ge Li
  • Peng Jiang
  • Ruo Dan Liu
  • Xuan Yang
  • Li Na Liu
  • Pei Liu
  • Shuai Bing Zhang
  • Zhong Quan Wang
Short Communication

Abstract

Glutathione-S-transferase (GST) is a family of multifunctional enzymes catalyzing detoxification reactions. Our previous study showed that Trichinella spiralis GST (TsGST) gene is an up-regulated gene in intestinal infective larvae (IIL) compared to muscle larvae (ML) and vaccination of mice with rTsGST displayed a partial protection against challenge infection. The purified rTsGST showed the maximum enzymatic activity at pH 6.5 and 40 °C. The enzymatic K m values for GSH and CDNB were 457 and 123 μM, respectively. An in vitro invasion assay showed that when anti-rTsGST serum of mice infected with T. spiralis and normal mouse serum were added to the medium, and the invasion rate of the infective larvae in an intestinal epithelial cell (IEC) monolayer was 31.0, 11.36, and 78.96 %, respectively (P < 0.05), which indicates that anti-rTsGST antibodies partially inhibited the larval invasion of IEC. ADCC assay showed that anti-rTsGST serum induced significant death of larvae (70 % cytotoxicity) compared to the larvae incubated with pre-immune serum (12 % cytotoxicity, P < 0.001) and was dose dependent.

Keywords

Trichinella spiralis Glutathione-S-transferase Enzyme activity ADCC Invasion 

Notes

Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 81271860 and 81471981).

References

  1. Armstrong RN (1997) Structure, catalytic mechanism, and evolution of the glutathione transferases. Chem Res Toxicol 10(1):2–18CrossRefPubMedGoogle Scholar
  2. Boulanger D et al (1994) Vaccination of goats against the trematode Schistosoma bovis with a recombinant homologous schistosome-derived glutathione S-transferase. Parasite Immunol 16(8):399–406CrossRefPubMedGoogle Scholar
  3. Bruce RG (1970) Structure of the esophagus of the infective juvenile and adult Trichinella spiralis. J Parasitol 56(3):540–549CrossRefPubMedGoogle Scholar
  4. Capron A, Capron M, Dombrowicz D, Riveau G (2001) Vaccine strategies against schistosomiasis: from concepts to clinical trials. Int Arch Allergy Immunol 124(1–3):9–15CrossRefPubMedGoogle Scholar
  5. Corsini E, Liesivuori J, Vergieva T, Van Loveren H, Colosio C (2008) Effects of pesticide exposure on the human immune system. Hum Exp Toxicol 27(9):671–680CrossRefPubMedGoogle Scholar
  6. Cui J et al (2013) Proteomic analysis of surface proteins of Trichinella spiralis muscle larvae by two-dimensional gel electrophoresis and mass spectrometry. Parasit Vectors 6:355CrossRefPubMedCentralPubMedGoogle Scholar
  7. Despommier DD (1993) Trichinella spiralis and the concept of niche. J Parasitol 79(4):472–482CrossRefPubMedGoogle Scholar
  8. Forsberg L, de Faire U, Morgenstern R (2001) Oxidative stress, human genetic variation, and disease. Arch Biochem Biophys 389(1):84–93CrossRefPubMedGoogle Scholar
  9. Gagliardo LF, McVay CS, Appleton JA (2002) Molting, ecdysis, and reproduction of Trichinella spiralis are supported in vitro by intestinal epithelial cells. Infect Immun 70(4):1853–1859CrossRefPubMedCentralPubMedGoogle Scholar
  10. Grezel D, Capron M, Grzych JM, Fontaine J, Lecocq JP, Capron A (1993) Protective immunity induced in rat schistosomiasis by a single dose of the Sm28GST recombinant antigen: effector mechanisms involving IgE and IgA antibodies. Eur J Immunol 23(2):454–560CrossRefPubMedGoogle Scholar
  11. Gupta S, Bhandari YP, Reddy MV, Harinath BC, Rathaur S (2005) Setaria cervi: immunoprophylactic potential of glutathione-S-transferase against filarial parasite Brugia malayi. Exp Parasitol 109(4):252–255CrossRefPubMedGoogle Scholar
  12. Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem 249(22):7130–7139PubMedGoogle Scholar
  13. Jiang P, Wang ZQ, Cui J, Zhang X (2012) Comparison of artificial digestion and Baermann’s methods for detection of Trichinella spiralis pre-encapsulated larvae in muscles with low-level infections. Foodborne Pathog Dis 9(1):27–31CrossRefPubMedGoogle Scholar
  14. Laborde E (2010) Glutathione transferases as mediators of signaling pathways involved in cell proliferation and cell death. Cell Death Differ 17(9):1373–1380CrossRefPubMedGoogle Scholar
  15. Li F, Cui J, Wang ZQ, Jiang P (2010) Sensitivity and optimization of artificial digestion in the inspection of meat for Trichinella spiralis. Foodborne Pathog Dis 7(8):879–885CrossRefPubMedGoogle Scholar
  16. Liu RD, Wang ZQ, Wang L, Long SR, Ren HJ, Cui J (2013) Analysis of differentially expressed genes of Trichinella spiralis larvae activated by bile and cultured with intestinal epithelial cells using real-time PCR. Parasitol Res 112(12):4113–4120CrossRefPubMedGoogle Scholar
  17. ManWarren T, Gagliardo L, Geyer J, McVay C, Pearce-Kelling S, Appleton J (1997) Invasion of intestinal epithelia in vitro by the parasitic nematode Trichinella spiralis. Infect Immun 65(11):4806–4812PubMedCentralPubMedGoogle Scholar
  18. McVay CS, Tsung A, Appleton J (1998) Participation of parasite surface glycoproteins in antibody-mediated protection of epithelial cells against Trichinella spiralis. Infect Immun 66(5):1941–1945PubMedCentralPubMedGoogle Scholar
  19. Morrison CA et al (1996) Protection of cattle against Fasciola hepatica infection by vaccination with glutathione S-transferase. Vaccine 14(17–18):1603–1612CrossRefPubMedGoogle Scholar
  20. Moskwa B (1999) Trichinella spiralis: in vitro cytotoxicity of peritoneal cells against synchronous newborn larvae of different age. Parasitol Res 85(1):59–63CrossRefPubMedGoogle Scholar
  21. Preyavichyapugdee N, Sahaphong S, Riengrojpitak S, Grams R, Viyanant V, Sobhon P (2008) Fasciola gigantica and Schistosoma mansoni: vaccine potential of recombinant glutathione S-transferase (rFgGST26) against infections in mice. Exp Parasitol 119(2):229–237CrossRefPubMedGoogle Scholar
  22. Ren HJ, Cui J, Wang ZQ, Liu RD (2011) Normal Mouse Intestinal Epithelial Cells as a Model for the in vitro Invasion of Trichinella spiralis Infective Larvae. PLoS One 6(10):e27010CrossRefPubMedCentralPubMedGoogle Scholar
  23. Ren HJ, Cui J, Yang W, Liu RD, Wang ZQ (2013) Identification of differentially expressed genes of Trichinella spiralis larvae after exposure to host intestine milieu. PLoS One 8(6):e67570CrossRefPubMedCentralPubMedGoogle Scholar
  24. Sheehan D, Meade G, Foley VM, Dowd CA (2001) Structure, function and evolution of glutathione transferases: implications for classification of non-mammalian members of an ancient enzyme superfamily. Biochem J 360(1):1–16CrossRefPubMedCentralPubMedGoogle Scholar
  25. Tu CP, Akgul B (2005) Drosophila glutathione S-transferases. Methods Enzymol 401:204–226CrossRefPubMedGoogle Scholar
  26. Veerapathran A, Dakshinamoorthy G, Gnanasekar M, Reddy MVR, Kalyanasundaram R (2009) Evaluation of Wuchereria bancrofti GST as a Vaccine Candidate for Lymphatic Filariasis. Plos Negl Trop Dis 3(6):e457CrossRefPubMedCentralPubMedGoogle Scholar
  27. Venturiello SM, Malmassari SL, Costantino SN, Nunez GG (2000) Cytotoxicity-blocking antibodies in human chronic trichinellosis. Parasitol Res 86(9):762–767CrossRefPubMedGoogle Scholar
  28. Wang ZQ, Wang L, Cui J (2012) Proteomic analysis of Trichinella spiralis proteins in intestinal epithelial cells after culture with their larvae by shotgun LC-MS/MS approach. J Proteomics 75(8):2375–2383CrossRefPubMedGoogle Scholar
  29. Wang B, Wang ZQ, Jin J, Ren HJ, Liu LN, Cui J (2013) Cloning, expression and characterization of a Trichinella spiralis serine protease gene encoding a 35.5 kDa protein. Exp Parasitol 134(2):148–154CrossRefPubMedGoogle Scholar
  30. Yang Y et al (2001) Role of glutathione S-transferases in protection against lipid peroxidation. Overexpression of hGSTA2-2 in K562 cells protects against hydrogen peroxide-induced apoptosis and inhibits JNK and caspase 3 activation. J Biol Chem 276(22):19220–19230CrossRefPubMedGoogle Scholar
  31. Yang W et al (2015) Molecular identification and characterization of Trichinella spiralis proteasome subunit beta type-7. Parasit Vectors 8(1):18CrossRefPubMedCentralPubMedGoogle Scholar
  32. Zhan B et al (2005) Biochemical characterization and vaccine potential of a heme-binding glutathione transferase from the adult hookworm Ancylostoma caninum. Infect Immun 73(10):6903–6911CrossRefPubMedCentralPubMedGoogle Scholar
  33. Zhan B et al (2010) Molecular cloning, biochemical characterization, and partial protective immunity of the heme-binding glutathione S-transferases from the human hookworm Necator americanus. Infect Immun 78(4):1552–1563CrossRefPubMedCentralPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Jing Cui
    • 1
  • Ling Ge Li
    • 1
  • Peng Jiang
    • 1
  • Ruo Dan Liu
    • 1
  • Xuan Yang
    • 1
  • Li Na Liu
    • 1
  • Pei Liu
    • 1
  • Shuai Bing Zhang
    • 1
  • Zhong Quan Wang
    • 1
  1. 1.Department of Parasitology, Medical CollegeZhengzhou UniversityZhengzhouPeople’s Republic of China

Personalised recommendations