Volume 19, Issue 74 (3-2011)                   J Adv Med Biomed Res 2011, 19(74): 44-53 | Back to browse issues page

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Haniloo A, Najafi F, Fazaeli A, Nourian A A. Comparison and Evaluation of Echinococcus granulosus Protoscoleces Excretory/Secretory Proteins in PBS Complemented with Glucose, DMEM and RPMI Culture Media. J Adv Med Biomed Res 2011; 19 (74) :44-53
URL: http://journal.zums.ac.ir/article-1-1416-en.html
Comparison and Evaluation of Echinococcus granulosus Protoscoleces Excretory/Secretory Proteins in PBS Complemented with Glucose, DMEM and RPMI Culture Media
Ali Haniloo1 , Faezeh Najafi *2 , Asghar Fazaeli1 , Ali Abbas Nourian1
1- Dept. of Parasitology and Entomology, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
2- Dept. of Parasitology and Entomology, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran. , faezeh_najafi@yahoo.com
Abstract:   (167403 Views)

Background and Objective: Preparation of proper antigens is an important issue in serology of hydatidosis. Investigators have been able to obtain excretory/secretory antigens (E/S Ags) by short-term culture of protoscoleces in a couple of culture media. However, no data are available about production rate of such antigens in different culture media. The present study was carried out to evaluate the production of E/S Ags (proteins) in PBS complemented with glucose, DMEM and RPMI culture media.
Material and Methods: To obtain E/S proteins, protoscoleces of echinococcus were cultured in PBS complemented with 10% glucose, RPMI and DMEM for 72 hours. Proteins secreted in culture media were concentrated and assayed. To characterize different components, proteins were electrophoresed on SDS-PAGE. Data were analyzed using One-way ANOVA and Tukey HSD tests.
Results: The mean concentration of E/S proteins in PBS medium in 24 hours of culture was significantly higher than DMEM and RPMI (P<0.05). However, such a difference was not observed between E/S proteins in DMEM and RPMI media. E/S proteins obtained from PBS medium were separated into 12 major bands and the two other media into 14 major bands within a range of molecular masses of 16 to 67 kDa.
Conclusion: PBS complemented with glucose is more appropriate than the two other media for E/S proteins production. The best time to obtain E/S proteins is the first 48 hours of culture.

Keywords: Echinococcus granulosus, Protoscoleces, Excretory/Secretory antigens
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Type of Study: Original Research Article |
Received: 2011/03/8 | Accepted: 2014/06/23 | Published: 2014/06/23
References
1. Eckert J, Deplazes P. Biological epidemiological and clinical aspects of echinococcosis: A zoonosis of increasing concern. Clin Microbiol Rev. 2004; 17: 107-35. [DOI:10.1128/CMR.17.1.107-135.2004] [PMID] [PMCID]
2. Thompson RCA. Echinococcosis. In: Gillespie SH, Pearson RD, eds. Principles and Practice of Clinical Parasitology. England: John Wiley & Sons; 2000: 585-612. [DOI:10.1002/0470842504.ch22] [PMID]
3. Mamuti W, Sako Y, Nakao M, et al. Recent advances in characterization of Echinococcus antigen B. Parasitol Int. 2006; 55: S57-S62. [DOI:10.1016/j.parint.2005.11.008] [PMID]
4. Carmena D, Benito A, Eraso E. Antigens for the immunodiagnosis of Echinococcus granulosus infection: An update. Act Trop. 2006; 98: 74-86. [DOI:10.1016/j.actatropica.2006.02.002] [PMID]
5. Haniloo A, Massoud J, Rokni MB. Evaluation and comparison of antigen B-ELISA and antigen B-immunoblotting in immunodiagnosis of cystic hydatid disease. Pak J Med Sci. 2005; 21: 352-6.
6. Lightowlers MW, Gottstien B. Echinococcosis/Hydatidosis: antigens, immunological and molecular diagnosis. In: Thompson RCA, Lymbery AJ, eds. Echinococcus and Hydatid Disease. UK, Wallingford: CAB International; 1995: 355-410.
7. Rafiei A, Craig PS. The immunodiagnostic potential of protoscolex antigens in human cystic echinococcosis and the possible influence of parasite strain. Ann Trop Med parasitol. 2002; 94: 383-9. [DOI:10.1179/000349802125001195] [PMID]
8. Carmena D, Benito A, Martinez J, Guisantes JA. Preliminary study of the presence of antibodies against excretory-secretory antigens from protoscoleces of Echinococcus granulosus in dogs with intestinal echinococcosis. Mem Inst Oswaldo Cruz, Rio de Janeiro. 2005; 100: 311-7. [DOI:10.1590/S0074-02762005000300018] [PMID]
9. Carmena D, Martinez J, Benito A, Guisantes JA. Shared and non-shared antigens from three different extracts of the metacestode of Echinococcus granulosus. Mem Inst Oswaldo Cruz, Rio de Janeiro. 2005; 100: 861-7. [DOI:10.1590/S0074-02762005000800007] [PMID]
10. Carmena D, Benito A, Postigo I, Arteaga J, Martinez J, Guisantes JA. Short term culture of protoscoleces to obtain excretory-secretory proteins of Echinococcus granulosus. Res Rev Parasitol. 2002; 62: 84-8.
11. Gasser RB, Lightowlers MW, Rickard MD. Identification of protein components of Echinococcus granulosus protoscolex antigens for specific serodiagnosis of E. granulosus infection in dogs. Parasite Immunol. 1989; 11: 279-91. [DOI:10.1111/j.1365-3024.1989.tb00666.x] [PMID]
12. Zhang WB, Jones Mk, LI J, McManus DP. Echinococcus granulosus: Preculture of protoscoleces in vitro significantly increases development and viability of secondary hydatid cysts in mice. Exp Parasitol. 2005; 110: 88-90. [DOI:10.1016/j.exppara.2005.02.003] [PMID]
13. Carmena D, Martinez J, Benito A, Guisantes JA. Characterization of excretory-secretory products from protoscoleces of Echinococcus granulosus and evaluation of their potential for immunodiagnosis of human cystic echinococcosis. Parasitol. 2004; 129: 371-8. [DOI:10.1017/S0031182004005670] [PMID]
14. Haniloo A, Ghasemi F, sheikhi AK, Ghavami MB. Immunoregulatory cytokine (TGF-β and IL-10) responses in mice inoculated with protoscoleces and major hydatid fluid antigens of cystic echinococcosis. Iranian J parasitol. 2008; 3: 18-23.
15. Rabinson RD, Arme G. Echinococcus granulosus: Failure of the eosin exclusion test to demonstrate death of protoscoleces. Ann Trop Med Parasitol. 1985; 79: 117-23. [DOI:10.1080/00034983.1985.11811897] [PMID]
16. Bollag DM, Edelstien SJ. Protein methods. New York: Wiley-Liss Inc; 1991.
17. Noble ER, Noble GA, Schad GA, MacInnes AJ. Parasitology: The Biology of Animal Parasites. London: Lea and Febiger; 1989.
18. Jeffs SA, Arme C. Glucose transport in protoscoleces of Echinococcus granulosus (cestoda). Comparative biochemistry and physiology. 1988; 91: 203-7. [DOI:10.1016/0300-9629(88)91617-9]
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