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Parasitology Research

, Volume 108, Issue 3, pp 593–599 | Cite as

Protein change of intestinal epithelial cells induced in vitro by Trichinella spiralis infective larvae

  • Shu Wei Wang
  • Zhong Quan Wang
  • Jing Cui
Original Paper

Abstract

The aim of this study was to observe the protein changes of intestinal epithelial cells induced in vitro by Trichinella spiralis infective larvae and their excretory-secretory (ES) or surface antigens and identity the proteins related with invasion. HCT-8 cells were incubated for 2 h in the culture medium contained ES or surface antigens of infective larvae, and observed by Immunofluorescent test (IFT). The infective larvae were inoculated into culture of HCT-8 cells to incubate for 18 h, and the lysates of HCT-8 cells were analyzed by SDS-PAGE and Western blot. IFA showed that normal HCT-8 cells had positively reactions with sera of the infected mice and mice immunized with ES or surface antigens. However, after incubating with ES or surface antigens, HCT-8 cells had stronger positively reaction with the above sera. On Western blot, after cultured with infective larvae, additional seven protein bands (66, 61, 57, 45, 34, 21, and 17 kDa) of HCT-8 cells were recognized by sera of the infected or immunized mice, but three protein bands (48, 43, and 23 kDa) of HCT-8 cells were not recognized by the above sera, compared with normal HCT-8 cells. Our results showed that after cultured with infective larvae the protein components of HCT-8 cell changed, suggesting that additional seven proteins recognized by sera of the infected or immunized mice may be related with invasion of intestinal epithelial cells by infective larvae, these proteins might mediate or facilitate entry into the cells, while the three proteins not recognized by the above sera may be the specific mediators released from the cells which permit invasion.

Keywords

Surface Antigen Intestinal Epithelial Cell Infected Mouse Immunize Mouse Infective Larva 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This study was supported by the grants of the National Basic Research Program of China (No. 2010CB530000), the National Natural Science Foundation of China (No. 30972579), and Major Public Research Project of Henan Province (No. 2008-145).

Conflict of interest

None

References

  1. Ali Khan Z (1966) The postembryonic development of Trichinella spiralis with special reference to ecdysis. J Parasitol 52:248–259CrossRefGoogle Scholar
  2. Appleton JA, McGregor DD (1984) Rapid expulsion of Trichinella spiralis in suckling rats. Science 226:70–72CrossRefPubMedGoogle Scholar
  3. Appleton JA, Schain LR, McGregor DD (1988) Rapid expulsion of Trichinella spiralis in suckling rats: mediation by monoclonal antibodies. Immunology 65:487–492PubMedGoogle Scholar
  4. Bell RG, McGregor DD (1979) Trichinella spiralis: expression of rapid expulsion in rats exposed to an abbreviated enteral infection. Exp Parasitol 48:42–50CrossRefPubMedGoogle Scholar
  5. Bell RG, Appleton JA, Negrao-Correa DA, Adams LS (1992) Rapid expulsion of Trichinella spiralis in adult rats mediated by monoclonal antibodies of distinct IgG isotypes. Immunology 75:520–527PubMedGoogle Scholar
  6. Bolás-Fernandez F, Corral Bezara LD (2006) TSL-1 antigens of Trichinella: an overview of their potential role in parasite invasion, survival and serodiagnosis of trichinellosis. Res Vet Sci 81:297–303CrossRefPubMedGoogle Scholar
  7. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  8. Bruce RG (1970) Structure of the esophagus of the infective juvenile and adult Trichinella spiralis. J Parasitol 56:540–549CrossRefPubMedGoogle Scholar
  9. Bruschi F (2002) The immune response to the parasitic nematode Trichinella and the ways to escape it, from experimental studies to implications for human infection. Curr Drug Targets Immune Endocr Metabol Disord 2:269–280CrossRefPubMedGoogle Scholar
  10. Butcher BA, Gagliardo LF, ManWarren T, Appleton JA (2000) Larvae-induced plasma membrane wounds and glycoprotein deposition are insufficient for Trichinella spiralis invasion of epithelial cells. Mol Biochem Parasitol 107:207–218CrossRefPubMedGoogle Scholar
  11. Campbell WC (1983) Trichinella and trichinosis. Plenum Press, New YorkGoogle Scholar
  12. Cui J, Wang ZQ, Zhu W, Zhang RG (1999) Seroepidemiologicl study of Trichinella spiralis infection in central China. Helminthologia 36:235–239Google Scholar
  13. Cui J, Wang ZQ, Zhang D (2003) Study on specific diagnostic antigens in excretory–secretory products from muscle larvae of Trichinella spiralis. Chin J Parasitol Parasit Dis 21:268–271, in ChineseGoogle Scholar
  14. Dea-Ayuela MA, Bolas-Fernandez F (1999) Trichinella antigens: a review. Vet Res 30:559–571PubMedGoogle Scholar
  15. Drake L, Korchev Y, Bashford L, Djamgoz M, Wakelin D, Ashall F, Bundy D (1994) The major secreted product of the whipworm, Trichuris, is a pore-forming protein. Proc Biol Sci 257:255–261CrossRefPubMedGoogle Scholar
  16. Dzik JM (2006) Molecules released by helminth parasites involved in host colonization. Acta Biochim Pol 53:33–64PubMedGoogle Scholar
  17. Ellis LA, Reason AJ, Morris HR, Dell A, Iglesias R, Ubeira FM, Appleton JA (1994) Glycans as targets for monoclonal antibodies that protect rats against Trichinella spiralis. Glycobiology 4:585–592CrossRefPubMedGoogle Scholar
  18. Gagliardo LF, McVay CS, Appleton JA (2002) Molting, ecdysis, and reproduction of Trichinella spiralis are supported in vitro by intestinal epithelial cells. Infect Immun 70:1853–1859CrossRefPubMedGoogle Scholar
  19. Hotez P, Cappello M, Hawdon J, Beckers C, Sakanari J (1994) Hyaluronidases of the gastrointestinal invasive nematodes Ancylostoma caninum and Anisakis simplex: possible functions in the pathogenesis of human zoonoses. J Infect Dis 170:918–926PubMedGoogle Scholar
  20. Inaba T, Sato H, Kamiya H (2003) Monoclonal IgA antibody-mediated expulsion of Trichinella from the intestine of mice. Parasitology 126:591–598CrossRefPubMedGoogle Scholar
  21. Kapel CM, Gamble HR (2000) Infectivity, persistence, and antibody response to domestic and sylvatic Trichinella spp. in experimentally infected pigs. Int J Parasitol 30:215–221CrossRefPubMedGoogle Scholar
  22. Kozek WJ (1971) The molting pattern in Trichinella spiralis. I. A light microscope study. J Parasitol 57:1015–1028CrossRefPubMedGoogle Scholar
  23. Li F, Cui J, Wang ZQ, Jiang P (2010) Factors affecting the sensitivity of artificial digestion and its optimization for inspection of Trichinella spiralis in meat. Foodborne Pathogens Dis 7:879–885CrossRefGoogle Scholar
  24. Mahannop P, Chaicumpa W, Setasuban P, Morakote N, Tapchaisri P (1992) Immunodiagnosis of human trichinellosis using excretory-secretory (ES) antigen. J Helminthol 66:297–304CrossRefPubMedGoogle Scholar
  25. 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 65:4806–4812PubMedGoogle Scholar
  26. Martinez J, Rodriguez-Caabeiro F (2005) Relationship between heat shock protein levels and infectivity in Trichinella spiralis larvae exposed to different stressors. Parasitol Res 97:213–218CrossRefPubMedGoogle Scholar
  27. Mclaren DJ (1976) Nematode sense organs. Adv Parasitol 14:195–265CrossRefPubMedGoogle Scholar
  28. McVay CS, Tsung A, Appleton J (1998) Participation of parasite surface glycoproteins in antibody-mediated protection of epithelial cells against Trichinella spiralis. Infect Immun 66:1941–1945PubMedGoogle Scholar
  29. McVay CS, Bracken P, Gagliardo LF, Appleton J (2000) Antibodies to tyvelose exhibit multiple modes of interference with the epithelial niche of Trichinella spiralis. Infect Immun 68:1912–1918CrossRefPubMedGoogle Scholar
  30. Nagano I, Wu Z, Takahashi Y (2009) Functional genes and proteins of Trichinella spp. Parasitol Res 104:197–207CrossRefPubMedGoogle Scholar
  31. Nöckler K, Reckinger S, Broglia A, Mayer-Scholl A, Bahn P (2009) Evaluation of a Western blot and ELISA for the detection of anti-Trichinella-IgG in pig sera. Vet Parasitol 163:341–347CrossRefPubMedGoogle Scholar
  32. Pozio E (2007) World distribution of Trichinella spp. infections in animals and humans. Vet Parasitol 149:3–21CrossRefPubMedGoogle Scholar
  33. Pritchard DI, Crawford CR, Duce IR, Behnke JM (1985) Antigen stripping from the nematode epicuticle using the cationic detergent cetyltrimethylammonium bromide (CTAB). Parasite Immunol 7:575–585CrossRefPubMedGoogle Scholar
  34. Robinson MW, Massie DH, Connolly B (2007) Secretion and processing of a novel multi-domain cystatin-like protein by intracellular stages of Trichinella spiralis. Mol Biochem Parasitol 151:9–17CrossRefPubMedGoogle Scholar
  35. Romaris F, Appleton JA (2001) Invasion of epithelial cells by Trichinella spiralis: in vitro observations. Parasite 8:S48–S50PubMedGoogle Scholar
  36. Wang ZQ, Cui J, Wei HY, Han HM, Zhang HW, Li YL (2006a) Vaccination of mice with DNA vaccine induces the immune response and partial protection against T. spiralis infection. Vaccine 24:1205–1212CrossRefPubMedGoogle Scholar
  37. Wang ZQ, Cui J, Xu BL (2006b) The epidemiology of human trichinellosis in China during 2000–2003. Acta Trop 97:247–251CrossRefPubMedGoogle Scholar
  38. Wang SW, Cui J, Wang ZQ, Wang L (2010) Effect of immune sera on the invasion of T. spiralis infective larvae into intestinal epithelial cells and their development in vitro. Chin J Parasitol Parasit Dis 28:81–85Google Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  1. 1.Department of Parasitology, Medical CollegeZhengzhou UniversityZhengzhouPeople’s Republic of China

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