Evaluation of antibody against hydatid fluid, protoscolex and adult worms of Echinococcus granulosus antigens by ELISA in mice

Document Type : Research Articles

Authors

Ferdowsi University of Mashhad

Abstract

Echinococcosis is one of the world’s relevant zoonoses, affecting both humans and their domestic animals and caused by infection with the metacestode stage of the dog tapeworm, Echinococcus granulosus. In the present study, twenty mice were randomly divided into 4 groups of 5. Each mouse in groups 1, 2 and 3 was immunized with 100 μg of hydatid cyst fluid protein, protoscolex and adult worms of E. granulosus proteins in 100 μl of phosphate-buffered saline which was emulsified with an equal volume of Freund’s complete adjuvant respectively. In group 4, mice were just immunized with above adjuvant in PBS. Mice were boosted four weeks after the first immunization with the same preparation and with Freund’s incomplete adjuvant. Three weeks later, each mouse was challenged with 2000 protoscolices intraperitoneally. Sera were collected before and after each immunization and serum antibodies were tested by ELISA. Results showed that the level of antibody in mice which were immunized with adult worms of E. granulosus on day 28 was roughly seven times higher than before immunization and was higher than hydatid cyst fluid and protoscolex. Level of antibody in mice immunized with adult worms of E. granulosus was also higher than in hydatid cyst fluid and protoscolex groups at day 49. The results of this article indicate that antigen of adult worms of E. granulosus can be used as a candidate for production of a vaccine.

Keywords


Ahmad, G., W. A. Nizami, and M. K.Saifullah. 2001. Analysis of potential antigens of protoscolices isolated from pulmonary and hepatic hydatid cyst of bubalus bubalis. Comp.Immunol.
Microbiol. Infec. Dis. 24,91-110.
Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem.
72,248-254.
Craig, P. S., M. T. Rogan, and M. Camposponce.2003. Echinococcosis: disease,detection and transmission. Parasitology.
127,5-20.
Daly, L., G. Bourke, and J. McGilvray. 1992.Interpretation and uses of medical statistic.[4] ed. Blackwell, London.
Eckert, J. and P. Deplazes. 2004. Biological,epidemiological, and clinical aspects of echinococcosis, a zoonosis of increasing
concern. Clin Microbiol Rev. 17(1),107-135.
Fotiadis, C., C. Sergiou, J. Kirou, T. G.Troupis, J. Tselentis, P. Doussaitou, V. G.Gorgoulis, and M. N. Sechas. 1999.Experimental Echinococcus infection in the mouse model: pericystic cellular immunity reaction and effects on the lymphoid organs of immunocompetent and thymectomized
mice. In. Vivo. . 13,541-546.
Fraize, M., Sarciron, ME., Azzouz, S., Issaadi,N., Bosquet, G., and Petavy, AF.. 2005.Immunogenicity of two Echinococcus
granulosus antigens EgA31 and EgTrp in mice. Parasitol Res. 96(2),113-120.
Hashemi Tabar, G. R., Razmi, G.R., and Naghibi, A. . 2005. Trials to induce protective immunity in mice and sheep by application of protoscolex and hydatid fluid antigen or whole body antigen of
Echinococcus granulosus. . J. Vet. Med. B.52,243-245.
Hashemi Tabar, G. R., Razmi, G.R., and Naghibi, A. . 2006. rotective immunity in mice with whole body of Echinococcus
granulosus. . I B J. . 10(1),51-55.
Heath, D. D. and S. B. Lawrence. 1996. Antigenic polypeptides of Echinococcus granulosus oncospheres and definition of
protective molecules. Parasite Immunol. .18,374-357.
Heath, D. D., S. B. Lawrence, and W. K.Yong. 1992. Echinococcus granulosus in sheep: transfer from ewe to lamb of "Arc 5" antibodies and oncosphere-killing activity,
but not protection. Int. J. Parasitol. .22,1017-1021.
Hernandez, A. and A. Nieto. 1994. Induction of protective immunity against murine secondary hydatidosis. Parasite Immunol.16,537-544.
Jenkins, D. J. and M. D. Rickard. 1986.Specificity of scolex and oncosphere antigens for the serological diagnosis of
taeniid cestode infections in dogs. . Aust Vet J. . 63,40-42.
Khaled, M., Al-Qaoud., K. Sami, and Abdel-Hafez. 2005. Humoral and cytokine response during protection of mice against
secondary hydatidosis caused by Echinococcus granulosus. Parasitol Res. .98,54-60.
Lahmar, S., H. Debbek, L. H. Zhang, D. P.McManus, A. Souissi, S. Chelly, and P. R.Torgerson. 2004. Transmission dynamics of
the Echinococcus granulosus sheep-dog strain (G1 genotype) in camels in Tunisia.Vet Parasitol. . 121(1-2),151-156.
Lin, R., Ding, JB., Lu, XM,, Wang, XF.,Arziguli, Wei, XL., Wang, Y., and Wen, H.Transient expression of Echinococcus
granulosus Eg95 DNA vaccine and induction of immune response in mice.
Molan, A. and I. Saeed. 1988. Protection of mice against Echinococcus granulosus by previous inoculation with protoscolices exposed to ultraviolet irradiation. Japanese
J Parasitol. . 37,203-208.
Moro, P. L., H. H. Garcia, A. E. Gonzales, J. J.Bonilla, M. Verastegui, and R. H. Gilman.2005. Screening for cystic echinococcosis in an endemic region of Peru using portable
ultrasonography and the enzyme-linked immunoelectrotransfer blot (EITB) assay.Parasitol Res. 96(4),242-246.
Severi, M. A., G. Ferragut, and A. Nieto. 1997.Antibody response of Echinococcus granulosus infected mice: protoscolex
specific response during infection is associated with decreasing specific IgG1/IgG3 ratio as well as decreasing
avidity. . Parasite Immunol. . 19(12),545-552.
Shi, Z., Y. Wang, Z. Li, Z. Li, Y. Bo, R. Ma,and W. Zhao. 2009. Cloning, expression,and protective immunity in mice of a gene
encoding the diagnostic antigen P-29 of Echinococcus granulosus. . Acta Biochim Biophys Sin (Shanghai). 41(1),79-85.
Siles-Lucas, M. M. and B. B. Gottstein. 2001.Molecular tools for the diagnosis of cystic and alveolar echinococcosis. Trop. Med.
Inte. Health. 6(6),463.
Wen, H. and P. S. Craig. 1994. IgG subclass responses in human cystic and alveolar echinococcosis. Am. J. Trop. Med. Hyg.
51,741-748.
Zhang, W., J. Li, and D. P. McManus. 2003a.Concepts in Immunology and Diagnosis of Hydatid Disease. Clin. Microbiol. Rev.16(1),18-36.
Zhang, W., H. You, J. Li, Z. Zhang, G. Turson,H. Aili, J. Wang, and D. P. McManus.2003b. Immunoglobulin profiles in a
murine intermediate host model of resistance for Echinococcus. granulosus infection. Parasite Immunol. 25(3),161-168.