In Vitro and In Vivo Potential of RH Strain of Toxoplasma gondii (Type I) in Tissue Cyst Forming.

  • Qasem Asgari Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
  • Hossein Keshavarz Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran AND Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran.
  • Saeedeh Shojaee Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
  • Mohammad Hossein Motazedian Dept. of Medical Parasitology and Mycology, Faculty of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
  • Mehdi Mohebali Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran AND Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran.
  • Ramin Miri Medicinal and Natural Products Chemistry Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
  • Davood Mehrabani Stem Cell and Transgenic Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
  • Mostafa Rezaeian Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran AND Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran.
Keywords: RH strain, Tissue cyst, Toxoplasma gondii, in vitro, in vivo

Abstract

 Background: Based on recent studies, there are controversial reports on the ca-pacity of tissue cyst forming of Toxoplasma gondii RH strain. In this study, the capac-ity was evaluated by in vivo and in vitro experiments. Methods: RH strain was subcutaneously inoculated to ten Wistar rats. After one month, their blood, brain, tongue and diaphragm were collected and evaluated by MAT, PCR, pathological and bioassay methods. The parasite was cultivated in the cell monolayer. To change to bradyzoite, the media pH was altered to 6.8. Biologi-cal aspect of the bradyzoites was evaluated by incubation in acidic pepsin and it's inoculation in ten BALB/c mice. Results: All rats showed antibodies to Toxoplasma at titers ≥1:320 but no DNA and tissue cyst were detected in the tissues. Following intraperitoneal inoculation of rats’ brain homogenate into BALB/c mice, no infection was established in none of the animals. During presence of cell culture, in acid media for a 3-5 days period, cyst-like structures were noticed when they were stained with PAS. The visible bradyzoites in the cysts that were incubated in acid pepsin medium were not able to kill any mice. Conclusion: This study confirmed that Iranian RH strain has lost the potential of tissue cyst forming in rats and bradyzoites cultivated in cell culture lost their resis-tance to acidic condition, so this strain can be a candidate for future vaccine re-searches.

References

Smith JE,ReduckNR.Toxoplasma gondii Strain Variation and Pathogenicity. In: Cary JW, Linz JE,Bhatnaga B (eds.), Microbial Foodborne Diseases: Mechanisms of Pathogenesis and Toxin Synthesis. Lancaster: Technnomic Pub-lishing; 2000.

Clementino MM, Souza MF, Andrade Neto VF. Seroprevalence and Toxoplasma gondii-IgG avidity in sheep from Lajes, Brazil. Vet Parasitol. 2007; 146: 199-203.

Jittapalapong S, Nimsupan B, Pinyopanuwat N, Chimnoi W, Kabeya H, Maruyama S. Seroprevalence of Toxoplasma gondii antibodies in stray cats and dogs in the Bangkok metropolitan area, Thailand. Vet Parasitol. 2007; 145: 138-141.

Garcia JL, Navarro IT, Vidotto O, Gennari SM, Machado RZ, da Luz Pereira AB, Sinhorini IL. Toxoplasma gondii: comparison of a rhoptry-ELISA with IFAT and MAT for antibody detection in sera of experimentally infected pigs. Exp Parasitol. 2006; 113: 100-105.

Gilot-Fromont E, Aubert D, Belkilani S, Hermitte P, Gibout O, Geers R, Villena I. Landscape, herd management and within-herd seroprevalence of Toxoplasma gondii in beef cattle herds from Champagne-Ardenne, France. Vet Parasitol.2009; 161: 36-40.

Dubey JP. Advances in the life cycle of Toxoplasma gondii. Int J Parasitol. 1998; 28: 1019-1024.

Tenter AM, Heckeroth AR, Weiss LM. Toxoplasma gondii: from animals to humans. Int J Parasitol. 2000; 30: 1217-1258.

Powell CC, Brewer M, Lappin MR. Detection of Toxoplasma gondii in the milk of experimentally infected lactating cats. Vet Parasitol. 2001;102: 29-33.

Subauste C. Animal models for Toxoplasma gondii infection. Cur Protocols Immunol. 2012; 96 (19): 1–23.

Zenner L, Estaquier J, Darcy F, Maes P, Capron A, Cesbron-Delauw MF. Protective immunity in the rat model of congenital toxoplasmosis and the potential of excreted-secreted antigens as vaccine components. Parasite Immunol.1999; 21: 261-272.

Ashburn D, Evans R, Chatterton JM, Joss AW, Ho-Yen DO. Strategies for detecting Toxoplasma immunity. Br J Biomed Sci. 2003; 60: 105-108.

Ashburn D, Chatterton JM, Evans R, Joss AW, Ho-Yen DO. Success in the Toxoplasma dye test. J Infec.2001; 42: 16-19.

Rodgers L, Wang X, Wen X, Dunford B, Miller R, Suzuki Y. Strains of Toxoplasma gondii used for tachyzoite antigens to stimulate spleen cells of infected mice in vitro affect cytokine responses of the cells in the culture. Parasitol Res.2005; 97: 332-335.

Sabin AB. Toxoplasmic encephalitis in children. J Am Med Assoc. 1941; 116: 801-807.

Hughes HP, Hudson L, Fleck DG. In vitro culture of Toxoplasma gondii in primary and established cell lines. Int J Parasitol. 1986; 16: 317-322.

Sibley LD, Boothroyd JC. Virulent strains of Toxoplasma gondii comprise a single clonal lineage. Nature.1992; 359: 82-85.

Kreier JP. Parasitic Protozoa. New York; Academic Press:1977.

FrenkelJK, Dubey JP, Hoff RL. Loss of stages after continuous passage of Toxoplasma gondii and Besnoitia jellisoni. J Protozool. 1976; 23: 421-424.

Villard O, Candolfi E, Ferguson DJ, Marcellin L, Kien T. Loss of oral infectivity of tissue cysts of Toxoplasma gondii RH strain to outbred Swiss Webster mice. Int J Parasitol. 1997; 27: 1555-1559.

Weiss LM, Kim K. The development and biology of bradyzoites of Toxoplasma gondii. Front Biosci. 2000; 5: D391-405.

Ferreira-da-Silva MF, Rodrigues RM, Ferreira de Andrade E, de Carvalho L, Gross U, Lüder CGK Barbosa HS. Spontaneous stage differentiation of mouse-virulent Toxoplasma gondii RH parasites in skeletal muscle cells: an ultrastructural evaluation. Mem Do Institut Osw Cru.2009; 104: 196-200.

DubeyJP, Shen SK, Kwok OC, Frenkel JK. Infection and immunity with the RH strain of Toxoplasma gondii in rats and mice. J Parasitol. 1999; 85: 657-662.

Djurkovic-Djakovic O, Nikolic A, Bobic B, Klun I, Aleksic A. Stage conversion of Toxoplasma gondii RH parasites in mice by treatment with atovaquone and pyrrolidine dithiocarbamate. Microb. Infect / Institut Pasteur.2005; 7: 49-54.

Zare F, Dalimi-Asl A, Ghaffarifar F. Detection of active Toxoplasma gondii (RH strain) in the different body tissues of experimentally infected rats. Modares J Med Sci. 2007; 9: 19-23.

Abdoli A, Dalimi A, Movahedin M. Impaired reproductive function of male rats infected with Toxoplasma gondii. Andrologia.2012; 44: 679-687.

Akins CK, Panicker S, Cunningham CL. Laboratory Animals in Research and Teaching: Ethics, Care, and Methods. Washington DC;APA Press: 2004.

Dubey JP, Desmonts G. 1987. Serological res-ponses of equids fed with Toxoplasma gondii oo-cysts. Equine Vet J. 48:1239-1243.

Jones CD, Okhravi N, Adamson P, Tasker S, Lightman S. Comparison of PCR detection methods for B1, P30, and 18S rDNA genes of T. gondii in aqueous humor. Invest Ophthalmol Vis Sci. 2000; 41: 634-644.

Dubey JP. Refinement of pepsin digestion method for isolation of Toxoplasmagondii from infected tissues. Vet Parasitol.1998; 74:75-77.

Dubey JP, Navarro IT, Sreekumar C, Dahl E, Freire RL, Kawabata HH, Vianna M. C, Kwok OC, Shen SK, Thulliez P, Lehmann T. Toxoplasma gondii infections in cats from Parana, Brazil: seroprevalence, tissue distribution, and biologic and genetic characterization of isolates. J Parasitol. 2004; 90: 721-726.

Dubey JP, Beattie CP. Toxoplasmosis of Animals and Man., Boca Raton FL; CRC Press:. 1988.

Hellbrugge TF, Dahme E, Hellbrugge FK. [Experimental animal observations on transplacental infection with Toxoplasma]. Zeitschrift Tropenmedizin Parasitologie.1953; 4: 312-322.

Hellbrugge T, Spiegler W, Grewing W. [Clinical, morphologic and serologic findings in gen-eralized toxoplasmosis in rats]. Zentralblatt fur Bakteriologie, Parasitenkunde, Infektions-krankheiten und Hygiene. 1. Abt. Medizinisch-hygienische Bakteriologie, Virus-forschung Parasitologie Originale.1956; 165: 495-506.

Benedetto N, Folgore A, Ferrara C, Galdiero M. Susceptibility to toxoplasmosis: correlation between macrophage function, brain cyst formation and mortality in rats. New Microbiologica.1996; 19: 47-58.

De Champs C, Imbert-Bernard C, Belme-guenai A, Ricard J, Pelloux H, Brambilla E, Ambroise-Thomas P. Toxoplasma gondii: in vivo and in vitro cystogenesis of the virulent RH strain. J Parasitol. 1997: 83, 152-155.

Saraeii M, Shahnazi M, Esmaeili rostaghi AR, Shojaee S, azhdari Zarmehri H, Safdari F. Lost of cystogenesis ability of Toxoplasma gondii RH strain in rat. 7th national and 2nd reginal congress of parasitology and parasitic diseases in Iran. 2010; Oct 19-21: P 135.

Dubey JP, Frenkel JK. Toxoplasmosis of rats: a review, with considerations of their value as an animal model and their possible role in epidemiology. Vet Parasitol. 1998; 77: 1-32.

McLeod R, Frenkel JK, Estes RG, Mack DG, Eisenhauer PB, Gibori G. Subcutaneous and intestinal vaccination with tachyzoites of Toxoplasma gondii and acquisition of immunity to peroral and congenital Toxoplasma challenge. J Immunol. 1988; 140: 1632-1637.

Soete M, Camus D, Dubremetz JF. Experimental induction of bradyzoite-specific antigen expression and cyst formation by the RH strain of Toxoplasma gondii in vitro. Exp Parasitol.1994; 78: 361-370.

Weiss LM, Laplace D,. Takvorian PM, Tanowitz HB, Cali A, Wittner M. A cell culture system for study of the development of Toxoplasma gondii bradyzoites. J Eukar Microbiol.1995; 42: 150-157.

Lyons RE, McLeod R, Roberts CW. Toxoplasma gondii tachyzoite-bradyzoite interco-nversion. Trends Parasitol. 2002; 18: 198-201.

Guimaraes EV, de Carvalho L, Barbosa HS. An alternative technique to reveal poly-saccharides in Toxoplasma gondii tissue cysts. Mem Do Institut Osw Cru 2003; 98: 915-917.

Dubey JP. Re-examination of resistance of Toxoplasma gondii tachyzoites and bradyzoites to pepsin and trypsin digestion. Parasitol.1998; 116: 43-50.

Asgari Q, Mehrabani D, Motazedian MH, Kalantari M, Nouroozi J, Adnani Sadati SJ. The Viability and Infectivity of Toxoplasma gondii Tachyzoites in Dairy Products undergoing Food Processing. Asian J Anim Sci. 2011; 5: 202-207.

Lane A, Soete M, Dubremetz JF, Smith JE. Toxoplasma gondii: appearance of specific markers during the development of tissue cysts in vitro. Parasitol Res. 1996; 82: 340-346.

Popiel I, Gold MC, Booth KS. Quantification of Toxoplasma gondii bradyzoites. J Parasitol. 1996; 82: 330-332.

Bohne W, Heesemann J, Gross U. Induction of bradyzoite-specific Toxoplasma gondii anti-gens in gamma interferon-treated mouse mac-rophages. Infect Immun.1993; 61: 1141-1145.

Ferguson DJ, Hutchison WM. An ultrastructu-ral study of the early development and tissue cyst formation of Toxoplasma gondii in the brains of mice. Parasitol Res. 1987; 73: 483-491.

Yang S, Parmley SF. Toxoplasma gondii expresses two distinct lactate dehydrogenase homologous genes during its life cycle in intermediate hosts. Gene.1997; 184: 1-12.

Dzierszinski F, Mortuaire M, Dendouga N, Popescu O, TomavoS. Differential expression of two plant-like enolases with distinct enzymatic and antigenic properties during stage conversion of the protozoan parasite Toxoplasma gondii. J Mol Biol.2001; 309: 1017-1027.

Dzierszinski F, Popescu O, Toursel C, Slomianny C, Yahiaoui B, Tomavo S. The protozoan parasite Toxoplasma gondii expresses two functional plant-like glycolytic enzymes. Implications for evolutionary origin of api-complexans. J Biol Chem.1999; 274: 24888-24895.

Lekutis C, Ferguson DJ, Boothroyd JC. Toxoplasma gondii: identification of a deve-lopmentally regulated family of genes related to SAG2. Exp Parasitol. 2000; 96: 89-96.

Selseleh M, Modarressi MH, Mohebali M, ShojaeeS, Eshragian MR, Azizi E, Keshavarz H. Real-Time RT-PCR on SAG1 and BAG1 gene expression during stage conversion in immunosuppressed mice infected with Toxoplasma gondii Tehran Strain. Korean J Parasitol.2012; 50: 199-205.

Selseleh M, Modarressi MH, Mohebali M, Shojaee S, Eshragian MR, Selseleh M, Azizi E, Keshavarz H. Real-Time RT-PCR on SAG1 and BAG1 Gene Expression during Stage Conver-sion in Immunosuppressed Mice Infected with Toxoplasma gondii Tehran Strain. Korean J Parasitol. 2012; 50(3): 199–205.

How to Cite
1.
Asgari Q, Keshavarz H, Shojaee S, Motazedian MH, Mohebali M, Miri R, Mehrabani D, Rezaeian M. In Vitro and In Vivo Potential of RH Strain of Toxoplasma gondii (Type I) in Tissue Cyst Forming. IJPA. 8(3):367-75.
Section
Articles