Brain Tissue Cysts in Infected Mice with RH-Strain of Toxoplasma gondii and Evaluation of BAG1 and SAG1 Genes Expression
-
Monavar Selseleh
,
Mh Modarressi
,
S Shojaee
,
M Mohebali
,
Mr Eshraghian
,
Mina Selseleh
,
H Keshavarz
Abstract
Background: Toxoplasma gondii is an obligate intracellular parasite that infects humans at high preva-lence rates. The virulent RH strain of T. gondii is generally considered to have lost its cyst forming ca-pacity. This study performed to obtain tissue cysts in mice infected with tachyzoites of RH strain treated with sulfadiazine (SDZ). It provides the opportunity to analyze the conversion of tachyzoite to bradyzoite stage of the RH strain, followed by stage-specific gene-expression analyzing.
Methods: Two groups of Swiss-Webster and BALB/c mice were infected subcutaneously with 104 tachyzoites of T. gondii, RH strain and given SDZ (300 mg/l) with NaHCO3 (5 g-1) in drinking water from day1 to day 14 post infection (p.i). The infected mice were sacrificed on day 50 post infection. Their brains were removed and the numbers of tissue cysts were microscopically counted. Total RNA was extracted from brains and cDNA synthesis was carried out. Finally, RT-PCR (Reverse tran-scription PCR) was used to detect the expression of bradyzoite (BAG1) and tachyzoite (SAG1) specif-ic genes during tachyzoite / bradyzoite stage conversion.
Results: Sixty five percent of all infected mice were survived. Cysts were detectable in mice brain (45%) on day 50 p.i. Also RT-PCR of the brain samples was positive for SAG1 and BAG1.
Conclusion: It seems that conversion of tachyzoites to bradyzoites in brain of mice undergoing SDZ was not completed until 50 days after inoculation.
Remington JS, Mcleod R, Desmonts G. Tox-oplasmosis. In: Remington JS, and Klein JO: Infectious diseases of the fetus and newborn infant. The WB Saunders Co, Philadelphia.4th ed.1995; p.140-267.
Mariuz P, Steigbigel RT. Toxoplasma infection in HIV-infected patients. In: Joynson, DHM, Wreghitt TG. (Eds.), Toxoplasmosis: A comprehen-sive clinical guide. Cambridge Univer-sity Press, Cambridge, K. 2001.
Joseph P, Calderon MM, Gilman RH, Quispe ML, Cok J, Ticona E, Chavez V, Jimenez JA, Chang MC, Lopez MJ, Ev-ans CA. Optimization and evalua-tion of a PCR assay for detecting toxoplasmic encephalitis in pa-tients with AIDS. J Clin Microbiol. 2002; 40: 4409- 4503.
Fuentes I, Rubio J. Genotypic characterization of Toxoplasma strains associated with human toxoplasmosis in Spain: Direct analysis from clinical samples. J Clin Microbiol. 2001; 39 (4):1566-1570
Boothroyd JC, Grigg ME. Population biology of Toxoplasma gondii and its relevance to human infection: do different strains cause different disease? Current Opinion in Microbiol. 2002; 5:438- 442.
Sibley LD, Boothroyd JC. Virulent strains of Toxoplasma gondii comprise a single clonage li-neage. Nature. 1992; 359: 82- 85.
Howe Dk, Sibley LD. Toxoplasma gondii com-prise three clonal lineage: correlation of pa-rasite genotype with human disease. J Infect Dis. 1995; 172: 1561-1566.
Dubey JP, Lindsay DS, Speer CA. Structures of Toxoplasma gondii tachyzoites, bradyzoites, and sporozoites and biology and development of tissue cysts. Clin Microbiol Rev. 1998; 11:267- 299.
Frenkel JK, Dubey JP, Hoff RL, Loss of stage after continuous passage of Toxoplasma gondii and Besnoitia jellisoni. J Protozool. 1996; 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 out bred Swiss-Webster mice. Int J Parasitol. 1997; 27: 1555-1559.
Clombo FA, Vidal JE, Penalava AC, Hernandez AV, Bonasser F, Nogue-ria RS. Diagnosis of cerebral toxoplas-mosis in AIDS patients: importance of molecular and immunological methods using peripheral blood sam-ples. J Clin Microbiol. 2005; 43:5044-5047.
Tomavo S, Weiss LM. Toxoplasma gene regula-tion and bradyzoite development. Mol Cell Biol. 2007; 285- 301.
Velmurugan GV, Tewari AK. High level ex-pression of SAG1 and GRA7 gene of Toxop-lasma gondii (Izantnagar isolate) and their appli-cation in serodiagnosis of goat tox-oplasmosis.Vet Parasitol. 2008; 154:185-192.
Lekutis C, Ferguson DJ, Grigg ME, Camps M, Boothroyd JC. Surface antigens of Toxoplasma gondii: variations on a theme. Int J Parasitol. 2001; 31:1285-1292.
Charles E, Callegan MC, Blader IJ. The SAG1 Toxoplasma gondii surface protein is not re-quired for acute ocular toxoplasmosis in mice. Infect and Immun. 2007; 75: 2079- 2083.
Ueno A, Dautu G. Toxoplasma gondii: identifi-cation and characterization of bradyzoite-specific deoxy ribose phosphate aldolase- like gene. (Tg DPA). Exp Parasitol. 2009; 121:55-63.
Wiess LM, Kim K. The development and bi-ology of bradyzoite of Toxoplasma gondii. Front Bioscience. 2000; D391- D405.
Ferguson DJ. Use of molecular and ultra structural markers to evaluate stage conversion of Toxoplasma gondii in both the intermediate and definitive host. Inter J Parasitol. 2004; 34, 347- 360
Gross U, Holpert M, Goebel S. Impact of stage differentiation on diagnosis of toxoplasmosis. Ann Ist Super Sanità. 2004; 40: 65–70.
Ferguson DJ, Parmley SF. Toxoplasma gondii MAG1 protein expression. Trends Parasitol. 2002; 18: 482- 485.
Ole A. Toxoplasmosis. In: OIE Manual of di-agnostic tests and vaccines for terrestrial ani-mals. 2008 ; 2. p.1286
Bohne W, Heesemann J, Gross U. Co- exis-tence of heterogeneous populations of Toxop-lasma gondii parasites with in parasitophorous vacuoles of murine macrophages as revealed by a bradyzoite- specific monoclonal antibody. Parasitol Res. 1993; 79: 485- 487.
Ferreira S, Barbosa HS, Groß U. Stress-related and spontaneous stage differentiation of Toxo-plasma gondii. Mol Biol. 2008; 4: 824- 834.
Lecomte V, Chumpitazi V, pasquier B, Am-broise TP. Brain - tissue cysts in rats infected with the RH strain of Toxoplasma gondii. Parasitol Res. 1992; 78: 267- 269.
Yano K, Nakabayashi T. Attenuation of the virulent RH strain of Toxoplasma gondii by pas-sages in mice immunized with Toxoplasma ly-sate antigens. Biken Journal.1986; 29: 31-37.
Dubey JP. Toxoplasma, Hammondia, Besnoita, Sarcocystis, and other tissue cyst -forming coc-cidian of man and animals. Parasitic Protozoa. 1988; 3: 101- 237.
Eliss J, Sinclair D. Microarrays and stage con-version in Toxoplasma gondii. Trends Parasitol. 2004; 20(6): 288- 294.
Buchbinder S, Blatz R. Comparison of real- time PCR detection methods for b1 and p30 genes of Toxoplasma gondii. Diag Microbiol Infec Dis. 2003; 45(4): 269- 271.
Cultrera R, Seraceni S, Segala D. Expression of toxoplasmic 65 KDa cystic mRNA by RT-PCR in patients with Toxoplasma gondii in-fection relapses. J Eukaryot Microbiol. 2001; 193-195
Switaj K, Master A, Skrzypczak M. Recent trends in molecular diagnostics for Toxoplasma gondii infections. Clin Microbiol Infec. 2005; 11: 170- 176.
| Files | ||
| Issue | Vol 8 No 1 (2013) | |
| Section | Articles | |
| Keywords | ||
| BAG1 RH strain RT-PCR SAG1 Toxoplasma gondii | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
