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Behaviour of Toxoplasma gondii RH Ankara strain tachyzoites during continuous production in various cell lines

Published online by Cambridge University Press:  01 December 2005

M. DÖŞKAYA
Affiliation:
Department of Parasitology, Ege University Medical Faculty, 35100, Bornova/Izmir, Turkey
A. DEGˇİRMENCİ
Affiliation:
Department of Parasitology, Ege University Medical Faculty, 35100, Bornova/Izmir, Turkey
C. ÇİÇEK
Affiliation:
Department of Clinical Microbiology, Ege University Medical Faculty, 35100, Bornova/Izmir, Turkey
M. AK
Affiliation:
Department of Parasitology, Ege University Medical Faculty, 35100, Bornova/Izmir, Turkey
M. KORKMAZ
Affiliation:
Department of Parasitology, Ege University Medical Faculty, 35100, Bornova/Izmir, Turkey
Y. GÜRÜZ
Affiliation:
Department of Parasitology, Ege University Medical Faculty, 35100, Bornova/Izmir, Turkey
A. ÜNER
Affiliation:
Department of Parasitology, Ege University Medical Faculty, 35100, Bornova/Izmir, Turkey

Abstract

Toxoplasma gondii is an obligate intracellular protozoan parasite. The objective of the present study was to examine the behaviour of Toxoplasma gondii RH Ankara strain tachyzoites in a cell culture environment. The study represents the first step in determining whether T. gondii RH Ankara strain tachyzoites, grown in cell culture, are of sufficient quality to allow cessation of in vivo tachyzoite production for diagnostic assays. In the present study, T. gondii RH Ankara strain tachyzoites were continuously produced in myeloma X63.Ag8.653, HeLa, Hep-2, and Vero cell cultures for 2 months. The average size of the tachyzoites was 3×5·7 μm prior to the first inoculation but after continuous production, a marked decrease was noted in average tachyzoite size. The smallest tachyzoite size, was 1×2·1 μm after 2 months, in myeloma cell cultures even though the yield of tachyzoites increased. With other cell cultures, tachyzoite yields were not as high as myeloma cell culture although decrease in size was less. The smallest decrease in tachyzoite size, averaging 2×3·8 μm after 2 months, was observed in tachyzoites produced in HeLa cell cultures. A virulence assay in small groups of BALB/c mice, using tachyzoites derived from cell cultures, was also conducted. The preliminary results of the virulence assay suggest that as the size of the tachyzoites decreased, the virulence in mice decreased. Future research will focus on the effect of the size of cell culture-derived T. gondii RH Ankara strain tachyzoites on the virulence, protein expression, and the reliability of diagnostic assays. Ultimately, the behaviour of tachyzoites from various T. gondii strains will be observed in cell culture to determine if size is altered.

Type
Research Article
Copyright
© 2005 Cambridge University Press

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References

REFERENCES

Ashburn, D., Evans, R., Chatterton, J. M. W., Joss, A. W. L. and Ho-Yen, D. O. ( 2000). Toxoplasma dye test using cell culture derived tachyzoites. Journal of Clinical Pathology 53, 630633.CrossRefGoogle Scholar
Braveny, I., Winter, W. and Disko, R. ( 1978). A method of mass cultivation of Toxoplasma gondii in cell culture. Tropenmedizin und Parasitologie 29, 432434.Google Scholar
Buckley, S. M. ( 1973). Survival of Toxoplasma gondii in mosquito cell lines and establishment of continuous infection in Vero cell cultures. Experimental Parasitology 33, 2326.CrossRefGoogle Scholar
Chatterton, J. M. W., Evans, R., Ashburn, D., Joss, A. W. L. and Ho-Yen, D. O. ( 2002). Toxoplasma gondii in vitro culture for experimentation. Journal of Microbiological Methods 51, 331335.CrossRefGoogle Scholar
De Meerschman, F., Rettigner, C., Focant, C., Boreux, R., Pinset, C., Leclipteux, T. and Losson, B. ( 2002). Use of a serum-free medium to produce in vitro Neospora caninum and Toxoplasma gondii tachyzoites on Vero cells. Veterinary Research 33, 159168.CrossRefGoogle Scholar
Domzig, W., Seguela, J. P. and Binz, H. ( 1993). A method to obtain large quantities of Toxoplasma gondii tachyzoites with extreme purity. Journal of Parasitology 79, 613615.CrossRefGoogle Scholar
Dubey, J. P., Shen, S. K., Kwok, O. C. H. and Frenkel, J. K. ( 1999). Infection and immunity with the RH strain of Toxoplasma gondii in rats and mice. Journal of Parasitology 85, 657662.CrossRefGoogle Scholar
Evans, R., Chatterton, J. M. W., Ashburn, D., Joss, A. W. L. and Ho-Yen, D. O. ( 1999). Cell-culture system for continuous production of Toxoplasma gondii tachyzoites. European Journal of Clinical Microbiology and Infectious Diseases 18, 879884.CrossRefGoogle Scholar
Grimwood, B. G., Hechemy, K. and Stevens, R. W. ( 1979). Toxoplasma gondii: purification of trophozoites propagated in cell culture. Experimental Parasitology 48, 282286.CrossRefGoogle Scholar
Hassl, A. and Aspock, H. ( 1990). A rapid and simple method of purification of Toxoplasma gondii trophozoites originating from tissue culture for use in the indirect immunofluorescent antibody test. Zentralbl Bakteriol. 272, 509513.CrossRefGoogle Scholar
Hermentin, K. and Aspock, H. ( 1987). Higher yields and increased purity of in vitro grown Toxoplasma gondii. Zentralblatt für Bakteriologie Mikrobiologie und Hygiene 267, 272276.CrossRefGoogle Scholar
Howe, D. K. and Sibley, L. D. ( 1994). Toxoplasma gondii: analysis of different laboratory stocks of the RH strain reveals genetic heterogeneity. Experimental Parasitology 78, 242245.CrossRefGoogle Scholar
Hughes, H. P. A., Hudson, L. and Fleck, D. G. ( 1986). In-vitro culture of Toxoplasma gondii in primary and established cell lines. International Journal for Parasitology 16, 317322.CrossRefGoogle Scholar
Mavin, S., Evans, R., Chatterton, J. M., Ashburn, D., Joss, A. W. and Ho-Yen, D. O. ( 2003). Toxoplasma gondii from liquid nitrogen for continuous cell culture: methods to maximise efficient retrieval. British Journal of Biomedical Science 60, 217220.CrossRefGoogle Scholar
Mavin, S., Joss, A. W., Ball, J. and Ho-Yen, D. O. ( 2004). Do Toxoplasma gondii RH strain tachyzoites evolve during continuous passage? Journal of Clinical Pathology 57, 609611.Google Scholar
Montoya, J. G. and Liesenfeld, O. ( 2004). Toxoplasmosis. Lancet 363, 19651976.CrossRefGoogle Scholar
Obwaller, A., Hassl, A., Picher, O. and Aspock, H. ( 1995). An enzyme-linked immunosorbent assay with whole trophozoites of Toxoplasma gondii from serum-free tissue culture for detection of specific antibodies. Parasitology Research 81, 361364.CrossRefGoogle Scholar
Roos, D. S., Donald, R. G., Morrissette, N. S. and Moulton, A. L. ( 1994). Molecular tools for genetic dissection of the protozoan parasite Toxoplasma gondii. Methods in Cell Biology 45, 2763.Google Scholar
Sibley, L. D. and Howe, D. K. ( 1996). The genetic basis of acute virulence in mice. In Toxoplasma gondii, Current Topics in Microbiology and Immunology ( ed. Gross, U.), pp. 1113. Springer, Berlin.