Advertisement

Parasitology Research

, Volume 104, Issue 2, pp 355–361 | Cite as

Blastocystis exhibits inter- and intra-subtype variation in cysteine protease activity

  • Haris Mirza
  • Kevin S. W. TanEmail author
Original Paper

Abstract

Blastocystis is an enteric protistan parasite of zoonotic potential and poorly understood pathogenesis. We have previously reported that Blastocystis cysteine proteases can degrade human secretory IgA and are also responsible for the induction of IL-8 response in colonic epithelial cells in vitro. Differences in virulence between Blastocystis subtypes have been reported recently in both animal models and clinical studies, although cellular mechanisms for these differences are currently unknown. Parasites such as Giardia intestinalis and Entamoeba histolytica have distinct virulent and non-virulent strains which may be attributable to variations in their cysteine proteases. In the present study, variations in cysteine protease activity was observed between avian (subtype 7) and rodent (subtype 4) isolates of Blastocystis with avian isolates exhibiting approximately two times higher peak cysteine protease activity than rodent isolates. Cysteine protease activity and parasite cell size varied over time within cultures of the same isolate. An association between parasite cell size and protease activity was observed.

Keywords

Protease Activity Cysteine Protease Azocasein Cell Size Distribution Cysteine Protease Activity 
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 work was supported by generous grants from the National Medical Research Council (NMRC/1071/2006) and Biomedical Research Council (BMRC/05/1/21/19/378). We are grateful to Dr. Manoj Kumar for technical help and helpful discussions. We also thank Ms. Ng Geok Choo, Ms. Yin Jing and Mr. Ramachandran for technical support.

References

  1. Buret AG (2007) Mechanisms of epithelial dysfunction in giardiasis. Gut 56:316–317CrossRefPubMedGoogle Scholar
  2. Chen XQ, Singh M, Ho LC, Moe KT, Tan SW, Yap EH (1997) A survey of Blastocystis sp. in rodents. Lab Anim Sci 47:91–94PubMedGoogle Scholar
  3. Clark CG (1997) Riboprinting: a tool for the study of genetic diversity in microorganisms. J Eukaryot Microbiol 44:277–283CrossRefPubMedGoogle Scholar
  4. Concha C, Monardes A, Even Y, Morin V, Puchi M, Imschenetzky M Genevière AM (2005) Inhibition of cysteine protease activity disturbs DNA replication and prevents mitosis in the early mitotic cell cycles of sea urchin embryos. J Cell Physiol 204:693–703CrossRefPubMedGoogle Scholar
  5. Coradi ST, Guimarães S (2006) Giardia duodenalis: protein substrates degradation by trophozoite proteases. Parasitol Res 99:131–136CrossRefPubMedGoogle Scholar
  6. Davis PH, Schulze J, Stanley S Jr (2007) Transcriptomic comparison of two Entamoeba histolytica strains with defined virulence phenotypes identifies new virulence factor candidates and key differences in the expression patterns of cysteine proteases, lectin light chains, and calmodulin. Mol Biochem Parasitol 151:118–128CrossRefPubMedGoogle Scholar
  7. DuBois KN, Abodeely M, Sajid M, Engel JC, McKerrow JH (2006) Giardia lamblia cysteine proteases. Parasitol Res 99:313–316CrossRefPubMedGoogle Scholar
  8. Dunn LA, Boreham PF, Stenzel DJ (1989) Ultrastructural variation of Blastocystis hominis stocks in culture. Int J Parasitol 19:43–56CrossRefPubMedGoogle Scholar
  9. Gantt SM, Myung JM, Briones MR, Li WD, Corey EJ, Omura S, Nussenzweig V, Sinnis P (1998) Proteasome inhibitors block development of Plasmodium spp. Antimicrob Agents Chemother 42:2731–2738PubMedGoogle Scholar
  10. Guimarães S, Sogayar MI, Franco MF (2003) Protease activity in Giardia duodenalis trophozoites of axenic strains isolated from symptomatic and asymptomatic patients. Mem Inst Oswaldo Cruz 98:77–81PubMedGoogle Scholar
  11. Hirata KK, Que X, Melendez-Lopez SG, Debonath A, Myers S, Herdman DS, Orozco E, Bhattacharya A, McKerrow JH, Reed SL (2007) A phagocytosis mutant of Entamoeba histolytica is less virulent due to deficient proteinase expression and release. Exp Parasitol 115:192–199CrossRefPubMedGoogle Scholar
  12. Hussein EM, Hussein AM, Eida MM, Atwa MM (2008) Pathophysiological variability of different genotypes of human Blastocystis hominis Egyptian isolates in experimentally infected rats. Parasitol Res 102:853–860CrossRefPubMedGoogle Scholar
  13. Jorgensen P, Tyers M (2004) How cells coordinate growth and division. Curr Biol 14:R1014–R1027CrossRefPubMedGoogle Scholar
  14. Kaneda Y, Horiki N, Cheng XJ, Fujita Y, Maruyama M, Tachibana H (2001) Ribodemes of Blastocystis hominis isolated in Japan. Am J Trop Med Hyg 65:393–396PubMedGoogle Scholar
  15. Kaya S, Cetin ES, Aridoğan BC, Arikan S, Demirci M (2007) Pathogenicity of Blastocystis hominis, a clinical reevaluation. Turkiye Parazitol Derg 31:184–187PubMedGoogle Scholar
  16. Leber AL (1999) Intestinal amoebae. Clin Lab Med 19:601–619, viiPubMedGoogle Scholar
  17. MacPherson DW, MacQueen WM (1994) Morphological diversity of Blastocystis hominis in sodium acetate-acetic acid-formalin-preserved stool samples stained with iron hematoxylin. J Clin Microbiol 32:267–268PubMedGoogle Scholar
  18. Moncada D, Keller K, Chadee K (2005) Entamoeba histolytica-secreted products degrade colonic mucin oligosaccharides. Infect Immun 73:3790–3793CrossRefPubMedGoogle Scholar
  19. Noël C, Dufernez F, Gerbod D, Edgcomb VP, Delgado- Wiscogliosi P, Ho LC, Singh M, Wintjens R, Sogin ML, Capron M, Pierce R, Zenner L, Viscogliosi E (2005) Molecular phylogenies of Blastocystis isolates from different hosts: implications for genetic diversity, identification of species, and zoonosis. J Clin Microbiol 43:348–355CrossRefPubMedGoogle Scholar
  20. Panaro M, Cianciulli A, Mitolo V, Mitolo C, Acquafredda A, Brandonisio O, Cavallo P (2007) Caspase-dependent apoptosis of the HCT-8 epithelial cell line induced by the parasite Giardia intestinalis. FEMS Immunol Med Microbiol 51:302–309CrossRefPubMedGoogle Scholar
  21. Puthia MK, Vaithilingam A, Lu J, Tan KSW (2005) Degradation of human secretory immunoglobulin A by Blastocystis. Parasitol Res 97:386–389CrossRefPubMedGoogle Scholar
  22. Puthia MK, Sio SW, Lu J, Tan KSW (2006) Blastocystis ratti induces contact-independent apoptosis, F-actin rearrangement, and barrier function disruption in IEC-6 cells. Infect Immun 74:4114–4123CrossRefPubMedGoogle Scholar
  23. Puthia MK, Lu J, Tan KSW (2008) Blastocystis ratti contains cysteine proteases that mediate interleukin-8 response from human intestinal epithelial cells in an NF-kappaB-dependent manner. Eukaryot Cell 7:435–443CrossRefPubMedGoogle Scholar
  24. Que X, Kim SH, Sajid M, Eckmann L, Dinarello CA, McKerrow JH, Reed SL (2003) A surface amebic cysteine proteinase inactivates interleukin-18. Infect Immun 71:1274–1280CrossRefPubMedGoogle Scholar
  25. Reed SL, Keene WE, McKerrow JH (1989) Thiol proteinase expression and pathogenicity of Entamoeba histolytica. J Clin Microbiol 27:2772–2777PubMedGoogle Scholar
  26. Rupeš I (2002) Checking cell size in yeast. Trends Genet 18:479–485CrossRefPubMedGoogle Scholar
  27. Sharma M, Hirata K, Herdman S, Reed S (1996) Entamoeba invadens: characterization of cysteine proteinases. Exp Parasitol 84:84–91CrossRefPubMedGoogle Scholar
  28. Sio S, Puthia MK, Lee AS, Lu J, Tan KSW (2006) Protease activity of Blastocystis hominis. Parasitol Res 99:126–130CrossRefPubMedGoogle Scholar
  29. Stark D, van Hal S, Marriott D, Ellis J, Harkness J (2007) Irritable bowel syndrome: a review on the role of intestinal protozoa and the importance of their detection and diagnosis. Int J Parasitol 37:11–20CrossRefPubMedGoogle Scholar
  30. Stensvold CR, Suresh GK, Tan KSW, Thompson RC, Traub RJ, Viscogliosi E, Yoshikawa H, Clark CG (2007) Terminology for Blastocystis subtypes—a consensus. Trends Parasitol 23:93–96CrossRefPubMedGoogle Scholar
  31. Tan KSW (2008) New insights on classification, identification and clinical relevance of Blastocystis spp. Clin Microbiol Rev (in press)Google Scholar
  32. Tan KSW, Singh M, Yap EH (2002) Recent advances in Blastocystis hominis research: hot spots in terra incognita. Int J Parasitol 32:789–804CrossRefPubMedGoogle Scholar
  33. Ustün S, Turgay N (2006) Blastocystis hominis and bowel diseases. Turkiye Parazitol Derg 30:72–76 (in Turkish)PubMedGoogle Scholar
  34. van der Heijden HM, Landman WJ (2008) In vitro effect of herbal products against Histomonas meleagridis. Vet Parasitol 154:1–7CrossRefPubMedGoogle Scholar
  35. Ward W, Alvarado L, Rawlings ND, Engel JC, Franklin C, McKerrow JH (1997) A primitive enzyme for a primitive cell: the protease required for excystation of Giardia. Cell 89:437–444CrossRefPubMedGoogle Scholar
  36. Wong KH, Ng GC, Lin RT, Yoshikawa H, Taylor MB, Tan KSW (2008) Predominance of subtype 3 among Blastocystis isolates from a major hospital in Singapore. Parasitol Res 102:663–670CrossRefPubMedGoogle Scholar
  37. Zhang Z, Wang L, Seydel KB, Li E, Ankri S, Mirelman D, Stanley SL Jr (2000) Entamoeba histolytica cysteine proteinases with interleukin-1 beta converting enzyme (ICE) activity cause intestinal inflammation and tissue damage in amoebiasis. Mol Microbiol 37:542–548CrossRefPubMedGoogle Scholar
  38. Zierdt CH, Swan JC (1981) Generation time and growth rate of the human intestinal parasite Blastocystis hominis. J Protozool 28:483–485PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Laboratory of Molecular and Cellular Parasitology, Department of Microbiology, Yong Loo Lin School of MedicineNational University of SingaporeKent RidgeSingapore
  2. 2.Infectious Diseases Programme, Life Science InstituteNational University of SingaporeKent RidgeSingapore

Personalised recommendations