Abstract
Long-term culturing of Toxoplasma gondii cysts was accomplished in vitro in association with murine astrocytes and intermittent additions of gamma interferon to the media. Phase-contrast microscopy was used to follow the stages of cyst development, and electron microscopy confirmed the presence of morphologic characteristics of T. gondii cysts. T. gondii cysts formed in vitro had a single trilaminar membrane during both intracellular and extracellular existence and contained amorphous electron-dense material either throughout the cyst or in a uniform layer under the trilaminar membrane. The bradyzoites were similar to previous descriptions of tachyzoites in vitro except that they were smaller and contained numerous electron-lucent vacuoles. Gamma interferon is not necessary for cyst formation, but it controls the division of tachyzoites and may allow cysts to remain for prolonged periods without rupturing. In vitro-cultivated T. gondii cysts will be useful for producing bradyzoite and cyst antigens and for measuring the effects of antimicrobial agents and immune modulators on the viability of intracystic T. gondii.
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- CARVER R. K., GOLDMAN M. Staining Toxoplasma gondii with fluorescein-labeled antibody. III. The reaction in frozen and paraffin sections. Am J Clin Pathol. 1959 Aug;32(2):159–164. doi: 10.1093/ajcp/32.2.159. [DOI] [PubMed] [Google Scholar]
- Conley F. K., Jenkins K. A. Immunohistological study of the anatomic relationship of toxoplasma antigens to the inflammatory response in the brains of mice chronically infected with Toxoplasma gondii. Infect Immun. 1981 Mar;31(3):1184–1192. doi: 10.1128/iai.31.3.1184-1192.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fontana A., Fierz W., Wekerle H. Astrocytes present myelin basic protein to encephalitogenic T-cell lines. Nature. 1984 Jan 19;307(5948):273–276. doi: 10.1038/307273a0. [DOI] [PubMed] [Google Scholar]
- Frenkel J. K., Nelson B. M., Arias-Stella J. Immunosuppression and toxoplasmic encephalitis: clinical and experimental aspects. Hum Pathol. 1975 Jan;6(1):97–111. doi: 10.1016/s0046-8177(75)80111-0. [DOI] [PubMed] [Google Scholar]
- GARNHAM P. C., BAKER J. R., BIRD R. G. Fine structure of cystic form of Toxoplasma gondii. Br Med J. 1962 Jan 13;1(5271):83–84. doi: 10.1136/bmj.1.5271.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
- HOGAN M. J., YONEDA C., FEENEY L., ZWEIGART P., LEWIS A. Morphology and culture of Toxoplasma. Arch Ophthalmol. 1960 Nov;64:655–667. doi: 10.1001/archopht.1960.01840010657006. [DOI] [PubMed] [Google Scholar]
- Hoff R. L., Dubey J. P., Behbehani A. M., Frenkel J. K. Toxoplasma gondii cysts in cell culture: new biologic evidence. J Parasitol. 1977 Dec;63(6):1121–1124. [PubMed] [Google Scholar]
- Jones T. C., Hirsch J. G. The interaction between Toxoplasma gondii and mammalian cells. II. The absence of lysosomal fusion with phagocytic vacuoles containing living parasites. J Exp Med. 1972 Nov 1;136(5):1173–1194. doi: 10.1084/jem.136.5.1173. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jones T. C., Len L., Hirsch J. G. Assessment in vitro of immunity against Toxoplasma gondii. J Exp Med. 1975 Feb 1;141(2):466–482. doi: 10.1084/jem.141.2.466. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kasper L. H., Currie K. M., Bradley M. S. An unexpected response to vaccination with a purified major membrane tachyzoite antigen (P30) of Toxoplasma gondii. J Immunol. 1985 May;134(5):3426–3431. [PubMed] [Google Scholar]
- Krahenbuhl J. L., Remington J. S. In vitro induction of nonspecific resistance in macrophages by specifically sensitized lymphocytes. Infect Immun. 1971 Oct;4(4):337–343. doi: 10.1128/iai.4.4.337-343.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Luft B. J., Naot Y., Araujo F. G., Stinson E. B., Remington J. S. Primary and reactivated toxoplasma infection in patients with cardiac transplants. Clinical spectrum and problems in diagnosis in a defined population. Ann Intern Med. 1983 Jul;99(1):27–31. doi: 10.7326/0003-4819-99-1-27. [DOI] [PubMed] [Google Scholar]
- Lunde M. N., Jacobs L. Antigenic differences between endozoites and cystozoites of Toxoplasma gondii. J Parasitol. 1983 Oct;69(5):806–808. [PubMed] [Google Scholar]
- MATSUBAYASHI H., AKAO S. MORPHOLOGICAL STUDIES ON THE DEVELOPMENT OF THE TOXOPLASMA CYST. Am J Trop Med Hyg. 1963 May;12:321–333. doi: 10.4269/ajtmh.1963.12.321. [DOI] [PubMed] [Google Scholar]
- Matuschka F. R. The effect of sulfomethoxypyrazine-pyrimethamine therapy on the fine structure of Toxoplasma gondii-cysts in the brain of Mastomys natalensis. Scanning and transmission electron microscopical investigations. Zentralbl Bakteriol Orig A. 1977 Jul;238(3):419–429. [PubMed] [Google Scholar]
- Murray H. W., Spitalny G. L., Nathan C. F. Activation of mouse peritoneal macrophages in vitro and in vivo by interferon-gamma. J Immunol. 1985 Mar;134(3):1619–1622. [PubMed] [Google Scholar]
- Pfefferkorn E. R. Interferon gamma blocks the growth of Toxoplasma gondii in human fibroblasts by inducing the host cells to degrade tryptophan. Proc Natl Acad Sci U S A. 1984 Feb;81(3):908–912. doi: 10.1073/pnas.81.3.908. [DOI] [PMC free article] [PubMed] [Google Scholar]
- REMINGTON J. S., JACOBS L., KAUFMAN H. E. Studies on chronic toxoplasmosis; the relation of infective dose to residual infection and to the possibility of congenital transmission. Am J Ophthalmol. 1958 Nov;46(5 Pt 2):261–268. [PubMed] [Google Scholar]
- Ruskin J., Remington J. S. Toxoplasmosis in the compromised host. Ann Intern Med. 1976 Feb;84(2):193–199. doi: 10.7326/0003-4819-84-2-193. [DOI] [PubMed] [Google Scholar]
- Sheffield H. G., Melton M. L. The fine structure and reproduction of Toxoplasma gondii. J Parasitol. 1968 Apr;54(2):209–226. [PubMed] [Google Scholar]
- Stahl W., Matsubayashi H., Akao S. Modification of subclinical toxoplasmosis in mice by cortisone, 6-mercaptopurine and splenectomy. Am J Trop Med Hyg. 1966 Nov;15(6):869–874. doi: 10.4269/ajtmh.1966.15.869. [DOI] [PubMed] [Google Scholar]
- WANKO T., JACOBS L., GAVIN M. A. Electron microscope study of Toxoplasma cysts in mouse brain. J Protozool. 1962 May;9:235–242. doi: 10.1111/j.1550-7408.1962.tb02611.x. [DOI] [PubMed] [Google Scholar]
- Werner H., Matuschka F. R., Brandenburg I. Structural changes of Toxoplasma gondii bradyzoites and cysts following therapy with sulfamethoxypyrazine-pyrimethamine: studies by light and electron microscopy. Consequences for chemotherapy. Zentralbl Bakteriol Orig A. 1979 Oct;245(1-2):240–253. [PubMed] [Google Scholar]