Development of 116 kDa Fraction for Detecting Experimental Toxoplasma gondii Infections in Mice.
AbstractBackground: Serological diagnosis of Toxoplasma gondii infection using crude antigens may not be more accurate. To increase the diagnostic potency of anti-gens, isolation of their immunogenic fractions could be useful. The current re-search adopted to obtain an affinity isolated fraction from RH strain using CNBr Sepharose 4B column coupled with infected mice sera helping in detec-tion of IgM and IgG of toxoplasmosis due to RH strain and other strains. Methods: The isolated fraction was characterized by SDS-PAGE. Moreover, the diagnostic potency of the fraction was assessed by indirect ELISA in mice experimentally infected with RH strain and two other local strains; one of sheep origin and the other of human origin.Results: The fraction was found to be consisted of a single band of 116 kDa compared with 17 bands ranged from 116 to 16 kDa associated with crude ex-tract. The fraction proved potent diagnostic potentials of acute and chronic mice toxoplasmosis. Where it was detected both IgM and IgG antibodies as early as two days and as late as 2 months post experimental infection with any of the three strains. The level of detected IgM and IgG by RH fraction was higher in mice infected with RH strain than with local strains except IgM due to sheep strain parasite.Conclusions: The 116 kDa fraction of T. gondii tachyzoites can be considered as a candidate in improving of serodiagnosisof Toxoplasma infections.
Buxton D, Maley SW, Wright SE, Rodger S, Bartley P. Toxoplasma gondii and ovine toxo-plasmosis: new aspects of an old story. Vet Parasitol. 2007; 149: 25–28.
Raeghi S, Akaberi A, Sedeghi S. Seropreva-lence of Toxoplasma gondii in Sheep, Cattle and Horses in Urmia North-West of Iran. Iranian J Parasitol. 2011; 6: 90–94. 3. Dubey JP, Jones JL. Toxoplasma gondii infection in humans and animals in United States. Int J Parasitol. 2008; 38: 1257–1278. 4. Jin S, Chang ZY, Ming X, Min CL, Wei H, Sheng LY, Hong GX. Fast Dipstick Dye Immunoassay for Detection of Immunoglobulin G (IgG) and IgM Antibodies of Human Toxoplasmosis. Clin Vaccine Immunol. 2005; 12: 198-201
Ghazy AA, Shaapan RM, Abdel-Rahman, EH. Comparative serological diagnosis of toxo-plasmosis in horses using locally isolated Toxo-plasma gondii. Vet Parasitol. 2007; 145: 31–36.
Hong S, Chong C, Lee K, Kim T, Hong Y. Maintained Seroprevalence of Toxoplasmosis among the Residents of Jeju Island, Korea. Korean J Parasitol. 2001; 49: 309-311.
Luo Q, Upadhya R, Zhang H, Madrid-Aliste C, Nieves E, et al., Analysis of the glyco-proteome of Toxoplasma gondii using lectin affinity chro-matography and tandem mass spectrometry. Microbes Infect. 2011; 13: 1199-210.
Gross U, Holpert M, Goebel G. Impact of stage differentiation on diagnosis of toxoplas-mosis. Ann Ist Super Sanita. 2004; 40: 65–70.
Ahn MH, Hyun KH, Kang JO, Min DY. Par-tially purified Toxoplasma gondii antigens by immunoaffinity chromatography. Korean J Parasitol. 1997; 35: 251-258.
Villavedra M, Battistoni, JJ, Rossi S, Carol H, Nieto A. Avidity analysis of the human im-mune response to a chitin binding protein of Toxoplasma gondii. Int J Parasitol. 2001; 31: 1087–1092.
Fatoohi AF, Cozon GJ, Ganzaio P, Mayencon M, Greenland T, et al. Heterogeneity in cellular and humoral immune responses against T. gondii antigen in humans. Clin Exp Immunol. 2004; 136: 535–541.
Abdel-Rahman EH, Ghazy AA, Shaapan RM. Prevalence of antibodies to Toxoplasma gondii in farm horses in Egypt. J Egypt Vet Med Assoc. 2005; 65: 99–112.
Conde de Felipe, MM, Molina JM, RodrÃguez-Ponce E, Ruiz A, GonzÃ¡lez JF. IGM and IGG response to 29-35-kDa Toxoplasma gondii protein fractions in experimentally infected goats. J Parasitol. 2007; 93: 701-703.
Johnson MA, McDonald PJ, Neoh HS. Kinet-ics of the growth of Toxoplasma gondii (RH strain) in mice. Int J Parasitol. 1979; 9: 55–56.
Shaapan RM, Ghazy AA. Isolation of Toxo-plasma gondii from horse meat in Egypt PJBS. 2007; 10, (1): 174-177.
Davis SW, Dubey JP. Mediation of immunity to Toxoplasma gondii oocysts shedding cats. J Parasitol. 1995; 81: 882-886.
Sharma SP, Dubey JP. Quantitative survival of T. gondii tachyzoites and bradyzoites in pepsin and trypsin solutions. Am J Vet Res. 1981; 42: 128-130.
Waltman WD, Dreesen DW, Prickett DM, Blue JL, Oliver DG. Enzyme-linked immunosorbent assay for the detection of tox-oplasmosis in swine: interpreting assay results and comparing with other serologic tests. Am J Vet Res. 1984; 45: 1719-1725.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin Phe-nol reagent. J Biol Chem. 1951; 193: 262–27.
Dubey, J.P., Beattie, C.P., 1988. Toxoplasmosis of Animals and Man. CRC Press Inc., Boca Ra-ton, FL. USA.
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacterio-phage T4. Nature, London, 1970; 227: 680-685
Wray W, Boulikas T, Wray VP, Hancock R. Silver staining of proteins in polyacrylamide gels. Anal Biochem. 1981; 118: 197-203.
Fagbemi BO, Obarisiaghon IO, Mbuh JV. Detection of circulating antigen in sera of Fasciola gigantica infected cattle with antibodies reactive with a Fasciola-specific 88 kDa antigen. Vet Parasitol. 1995; 58: 235–246.
Lind P, Haugegaard J, Wingstrand A, Henrisken SA. The time course of the specific antibody response by various ELISAs in pigs experimentally infected with Toxoplasma gondii. Vet Parasitol. 1997; 71: 1–15.
Hillyer GM, Soler De-Galanes M, Rodriguez-Perez, J, Bjorland J. Use of the falcon assay screening test-enzyme-linked immunosorbent assay (FAST-ELISA) and the enzyme-linked immunoelectrotransfer blot (EITB) to deter-mine the prevalence of human fascioliasis in the Bolivian Altiplano. Am J Trop Med Hyg. 1992; 46: 603–609.
Gong H, Kobayashi K, Sugi T, Takemae H, Kurokawa H, et al. A novel PAN/apple do-main-containing proteins from Toxoplasma gondii: characterization and receptor identifica-tion. PLoS One. 2012; 7: 1-10.
Angeloni MB, Silva NM, Castro AS, Gomes AO, Silva DAO. Apoptosis and S-phase of the cell cycle in BeWo trophoblastic and HeLa cells are differentially modulated by Toxoplasma gondii strain types. Placenta. 2009; 30: 785–791.
Khaminets A, Hunn J.P, Könen-Waisman S, Zhao YO, Preukschat D. Coordinated loading of IRG resistance GTPases on to the Toxoplas-ma gondii parasitophorous vacuole. Cellular Mi-crobiology. 2010; 12: 939–961.
Robben PM, Mordue DG, Truscott SM, Takeda K, Akira S. Production of IL-12 by macrophages infected with Toxoplasma gondii depends on the parasite genotype. J Immunol. 2004; 172: 3686–3694.
Ong YC, Boyle JP, Boothroyd JC. Strain-Dependent Host transcriptional Responses to Toxoplasma Infection Are Largely Conserved in Mammalian and Avian Hosts. PLoS One. 2011; 6: 1-11.
Yap GS, Shaw MH, Ling Y, Sher A. Genetic analysis of host resistance to intracellular path-ogens: lessons from studies of Toxoplasma gondii infection. Microbes Infect. 2006; 8: 1174–1178.
Gazzinelli RT, Wysocka M, Hieny S, Scharton-Kersten T, Cheever A. In the absence of en-dogenous IL-10, mice acutely infected with Toxoplasma gondii succumb to a lethal immune response dependent on CD4+ T cells and ac-companied by overproduction of IL-12, IFN-gamma and TNF- alpha. J Immunol. 1996; 157: 798–805.
Stumhofer JS, Laurence A, Wilson EH, Huang E, Tato CM. Interleukin 27 negatively regulates the development of interleukin 17-producing T helper cells during chronic inflammation of the central nervous system. Nat Immunol. 2006; 7: 937–945.
Laliberté J, Carruthers V. Host cell manipula-tion by the human pathogen Toxoplasma gondii. Cellular and Molecular Life Sciences. 2008; 65: 1900–1915.
Pollard AM, Knoll LJ, Mordue DG. The role of specific Toxoplasma gondii molecules in ma-nipulation of innate immunity. Trends Parasitol. 2009; 25: 491–494.
Dubey JP, Ferreira LR, Martins J, McLeod R. Oral oocyst-induced mouse model of toxo-plasmosis: effect of infection with Toxoplasma gondii strains of different genotypes, dose, and mouse strains (transgenic, out-bred, in-bred) on pathogenesis and mortality. Parasitology. 2010; 139: 1-13.
Smith DD, Frenkel JK. Immunological com-parison of 124 isolates of Toxoplasma gondii. Parasitol Res. 2003; 91: 332-337.