Advertisement

Parasitology Research

, Volume 109, Issue 2, pp 321–327 | Cite as

Proteaese activity of Blastocystis hominis subtype3 in symptomatic and asymptomatic patients

  • Dina Marie Abdel-Hameed
  • Omayma Mohamed Hassanin
Original Paper

Abstract

Despite accumulating evidence indicating that Blastocystis hominis is pathogenic and that cysteine proteases are involved in its pathogenesis, few researches discussed the protease activity of B. hominis genetic subtypes. Therefore, the present study aims to identify the underlying pathogenic role of the proteases of B. hominis subtype 3 at different molecular weights in correlation to gastrointestinal symptoms. Of 65 patients with various clinical presentations referred to our laboratory for stool examination, 26 (40%) were B. hominis positive by stool culture. Of 26 (group I) B. hominis patients, 18 (69.2%) were symptomatic (group IA) and 8(30.8%) were asymptomatic (group IB). Of 25 normal control group (group II), 5 (20%) were B. hominis positive. Subtype 3 was the only genotype recovered by polymerase chain reaction. Of 26 patients in group I, 19 (73.1%) were immunocompetent and 7 (26.9%) were immunocompromised. Protease activities of B. hominis subtype 3 were recognized at 32-kDa (46.2%), 39-kDa (7.7%), 120-kDa (38.5%), 140-kDa (11.5%), and 215-kDa (19.2%) bands in gelatin sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteases were recognized in 17 (94.4%) out of 18 symptomatic Blastocystis patients versus 2 (25.0%) out of 8 asymptomatic patients. Proteases at 32 kDa were reported in 61.1% of symptomatic versus 12.5% of asymptomatic patients. It was concluded that proteases of B. hominis genetic subtype 3, particularly those at 32 kDa, could be considered a virulence factor that is responsible for protein degradation and have a possible pathogenic role in host immune evasion.

Keywords

Irritable Bowel Syndrome Protease Activity Stool Culture Asymptomatic Group Genetic Subtype 
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 developed at the Diagnostic and Research Laboratory of Parasitic Diseases and the Molecular Department of Medical Research Center, Faculty of Medicine, Ain Shams University, Cairo, Egypt. We would like to thank Ahmed K. Emara who helped in preparing the samples and in reviewing the language.

References

  1. Abdel Hameed DM, Hassanin OM, Zuel-Fakkar NM (2010) Association of Blastocystis hominis genetic subtypes with urticaria. Parasitol Res (in press)Google Scholar
  2. Abe N (2004) Molecular and phylogenetic analysis of Blastocystis isolates from various hosts. Vet Parasitol 120:235–242PubMedCrossRefGoogle Scholar
  3. Böhm-Gloning B, Knobloch J, Walderich B (1997) Five subgroups of Blastocystis hominis from symptomatic and asymptomatic patients revealed by restriction site analysis of PCR-amplified 16 S-like rDNA. Trop Med Int Health 2:771–778PubMedCrossRefGoogle Scholar
  4. Boorom KF, Smith H, Nimri L, Viscogliosi E, Spanakos G, Parkar U, Li LH, Zhou XN, Ok UZ, Leelayoova S, Jones MS (2008) Oh my aching gut: irritable bowel syndrome, Blastocystis, and asymptomatic infection. BMC Parasit Vectors 1:40CrossRefGoogle Scholar
  5. Clark CG (2000) Cryptic genetic variation in parasitic protozoa. J Med Microbiol 49:489–491PubMedGoogle Scholar
  6. Dogruman-Al F, Kustimur S, Yoshikawa H, Tuncer C, Simsek Z, Tanyuksel M, Araz E, Boorom K (2009) Blastocystis subtypes in irritable bowel syndrome and inflammatory bowel disease in Ankara, Turkey. Mem Inst Oswaldo Cruz 104:724–727PubMedCrossRefGoogle Scholar
  7. Elwakil HS, Talaat RM (2009) Genetic analysis of Blastocystis hominis isolated from symptomatic and asymptomatic human hosts in Egypt. J Egypt Soc Parasitol 39:99–109PubMedGoogle Scholar
  8. Graczyk TK, Shiff CK, Tamang L, Munsaka F, Beitin AM, Moss WJ (2005) The association of Blastocystis hominis and Endolimax nana with diarrheal stools in Zambian school-age children. Parasitol Res 98:38–43PubMedCrossRefGoogle Scholar
  9. 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 102:853–860PubMedCrossRefGoogle Scholar
  10. Jones MS, Whipps CM, Ganac RD, Hudson NR, Boroom K (2009) Association of Blastocystis subtype 3 and 1 with patients from an Oregon community presenting with chronic gastrointestinal illness. Parasitol Res 104:341–345PubMedCrossRefGoogle Scholar
  11. 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
  12. Kaya S, Cetin ES, Aridogan BC, Arikan S, Demirci M (2007) Pathogenicity of Blastocystis hominis, a clinical reevaluation. Turk Parazitol Derg 31:184–187Google Scholar
  13. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  14. Leder K, Hellard ME, Sinclair MI, Fairley CK, Wolfe R (2005) No correlation between clinical symptoms and Blastocystis hominis in immunocompetent individuals. J Gastroenterol Hepatol 20(9):1390–1394PubMedCrossRefGoogle Scholar
  15. Leelayoova S, Siripattanapipong S, Thathaisong U, Naaglor T, Taamasri P, Piyaraj P, Mungthin M (2008) Drinking water: a possible source of Blastocystis spp. subtype 1 infection in schoolchildren of a rural community in central Thailand. Am J Trop Med Hyg 79:401–406PubMedGoogle Scholar
  16. Li LH, Zhang XP, Lv S, Zhang L, Yoshikawa H, Wu Z, Steinmann P, Utzinger J, Tong XM, Chen SH, Zhou XN (2007) Cross-sectional surveys and subtype classification of human Blastocystis isolates from four epidemiological settings in China. Parasitol Res 102:83–90PubMedCrossRefGoogle Scholar
  17. Lockwood BC, North MJ, Scott KI, Bremner AF, Coombs GH (1987) The use of a highly sensitive electrophoretic method to compare the proteinases of trichomonads. Mol Biochem Parasitol 24:89–95PubMedCrossRefGoogle Scholar
  18. Motazedian H, Ghasemi H, Sadjjadi SM (2008) Genomic diversity of Blastocystis hominis from patients in southern Iran. Ann Trop Med Parasitol 102:85–88PubMedCrossRefGoogle Scholar
  19. Noël C, Dufernez F, Gerbod D, Edgcomb VP, Delgado-Viscogliosi 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 43:348–355PubMedCrossRefGoogle Scholar
  20. Puthia MK, Vaithilingam A, Lu J, Tan KS (2005) Degradation of human secretory immunoglobulin A by Blastocystis. Parasitol Res 97:386–389PubMedCrossRefGoogle Scholar
  21. Puthia M, Sio SW, Lu J, Tan KS (2006) Blastocystis ratti induces contact-independent apoptosis, F-actin rearrangement, and barrier function disruption in IEC-6 cells. Infect Immun 74:4114–4123PubMedCrossRefGoogle Scholar
  22. Rayan HZ, Ismail OA, El Gayar EK (2007) Prevalence and clinical features of Dientamoeba fragilis infections in patients suspected to have intestinal parasitic infection. J Egypt Soc Parasitol 37:599–608PubMedGoogle Scholar
  23. Sio SWS, Puthia MK, Lee ASY, Lu J, Tan KSW (2006) Protease activity of Blastocystis hominis. Parasitol Res 99:126–130PubMedCrossRefGoogle Scholar
  24. Souppart L, Moussa H, Cian A, Sanciu G, Poirier P, El Alaoui H, Delbac F, Boorom K, Delhaes L, Dei-Cas E, Viscogliosi E (2009a) Subtype analysis of Blastocystis isolates from symptomatic patients in Egypt. Parasitol Res 106(2):505–511PubMedCrossRefGoogle Scholar
  25. Souppart L, Sanciu G, Cian A, Wawrzyniak I, Delbac F, Capron M, Dei-Cas E, Boorom K, Delhaes L, Viscogliosi E (2009b) Molecular epidemiology of human Blastocystis isolates in France. Parasitol Res 105(2):413–421PubMedCrossRefGoogle Scholar
  26. Stensvold CR, Nielsen HV, Mølbak K, Smith HV (2009) Pursuing the clinical significance of Blastocystis—diagnostic limitations. Trends Parasitol 25:23–29PubMedCrossRefGoogle Scholar
  27. Tan KS (2004) Blastocystis in humans and animals: new insights using modern methodologies. Vet Parasitol 126:121–144PubMedCrossRefGoogle Scholar
  28. Tan KSW (2008) New insights on classification, identification, and clinical relevance of Blastocystis spp. Clin Microbiol Rev 21:639–665PubMedCrossRefGoogle Scholar
  29. Tan KS, Singh M, Yap EH (2002) Recent advances in Blastocystis hominis research: hot spots in terra incognita. Int J Parasitol 32:789–804PubMedCrossRefGoogle Scholar
  30. Tan TC, Ong SC, Suresh KG (2009) Genetic variability of Blastocystis sp. isolates obtained from cancer and HIV/AIDS patients. Parasitol Res 105:1283–1286PubMedCrossRefGoogle Scholar
  31. Tasova Y, Sahin B, Koltas S, Paydas S (2000) Clinical significance and frequency of Blastocystis hominis in Turkish patients with hematological malignancy. Acta Med Okayama 54:133–136PubMedGoogle Scholar
  32. Upcroft JA, Dunn LA, Dommett LS, Healey A, Upcroft P, Boreham PFL (1989) Chromosomes of Blastocystis hominis. Int J Parasitol 19:879–883PubMedCrossRefGoogle Scholar
  33. Wong KH, Ng GC, Lin RT, Yoshikawa H, Taylor MB, Tan KS (2008) Predominance of subtype 3 among Blastocystis isolates from a major hospital in Singapore. Parasitol Res 102:663–670PubMedCrossRefGoogle Scholar
  34. Yakoob J, Jafri W, Jafri N, Khan R, Islam M, Beg MA, Zaman V (2004) Irritable bowel syndrome: in search of an etiology: role of Blastocystis hominis. Am J Trop Med Hyg 70:383–385PubMedGoogle Scholar
  35. Yoshikawa H, Wu Z, Kimata I, Iseki M, Ali IK, Hossain MB, Zaman V, Haque R, Takahashi Y (2004) Polymerase chain reaction based genotype classification among human Blastocystis hominis populations isolated from different countries. Parasitol Res 92:22–29PubMedCrossRefGoogle Scholar
  36. Zierdt CH (1991) Blastocystis hominis—past and future. Clin Microbiol Rev 4:61–79PubMedGoogle Scholar
  37. Zuckerman MJ, Watts MT, Ho H, Meriano FV (1994) Blastocystis hominis infection and intestinal injury. Am J Med Sci 308:96–101PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Dina Marie Abdel-Hameed
    • 1
  • Omayma Mohamed Hassanin
    • 2
  1. 1.Department of Parasitology, Faculty of MedicineAin Shams UniversityCairoEgypt
  2. 2.Clinical Pathology, Molecular Biology Department, Medical Research Center, Faculty of MedicineAin-Shams University HospitalsCairoEgypt

Personalised recommendations