Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Role of cytoskeleton in Host Cell Invasion by Intracellular Protozoa Toxoplasma gondii

  • Lee, Sook-Hwan (Department of Obstertrics and Gynecology, College of Medicine, Pochon Cha University) ;
  • Lee, Boo-Young (Department of Parasitology & Institute of Biomedical Science, Hanyang University, College of Medicine) ;
  • Min, Duk-Young (Department of Obstertrics and Gynecology, College of Medicine, Pochon Cha University) ;
  • Kim, Jung-Mogg (Department of Microbiology, Hanyang University, College of Medicine) ;
  • Ahn, Myoung-Hee (Department of Obstertrics and Gynecology, College of Medicine, Pochon Cha University)
  • Published : 2002.08.01
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Abstract

A microfilament-based motility in Toxoplasma gondii (T. gondii) Is involved in host cell invasion, yet the exact mechanism has not yet been determined. Accordingly, the current study examined the localization of actin and tubulin in T gondii using immunofluorescent (IF) and immunogold staining for electron microscopy. Indirect immunofluorescence (IF) staining using anti-actin and anti-tubulin monoclonal antibodies (mAbs) revealed localization of fluorescence on the entire surface of the tachyzoites. The actin in T. gondii was observed by immunogold staining, and the gold particles were seen on the surface, especially at the anterior end and in the cytoplasm of the parasite. However, there were no gold particles in the nucleus, rhoptries, and dense granules. The tubulin in T gondii was located on the surface and in the cytoplasm of the tachyzoites in the extracellular parasite, compared with anterior part of tachyzoites in the intracellular parasite. The antigens of T gondii recognized by anti-actin mAb were 107 kDa, 50 kDa, 48 kDa, and 40 kDa proteins, while those recognized by anti-tubulin mAb were 56 kDa, 52 kDa, and 34 kDa proteins. Tachyzoites of T gondii pretreated with the actin inhibitor, cytochalasin D (20 $\mu\textrm{g}$/ml), and tubulin inhibitor, colchicine (2$\times$10$\^$-6/ M), for 30 min at 37$\^{C}$ were used to infect the isolated mouse macrophages (tachyzo ites:macrophage=2:1). Pretreatment with the inhibitors resulted in lower multiplication of tachyzoites within the macrophages than in the untreated group 18 h post infection (p<0.05). Therefore, the present results suggest that actin and tubulin appear to be involved in the invasion of and multiplication in host cells.

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References

  1. J. Parasitol. v.84 Lack of therapeutic effect of colchicine on murine Toxoplamaosis Aguirre-Cruz, L.;J. Sotelo https://doi.org/10.2307/3284549
  2. J. Parasitol. v.82 Colchicine decreases the infection by Toxoplasma gondii in cultured glial cells Aguirre-Cruz, L.;M. Calderon;J. Sotelo https://doi.org/10.2307/3284170
  3. J. Protozool. v.36 Antigenic reactivity of monoclonal antibody against Amoeba proteus actin Ahn, T. I.;K. W. Jeon https://doi.org/10.1111/j.1550-7408.1989.tb01096.x
  4. J. Microbiol. Biotechnol. v.11 Production and characterization of monoclonal antibody to glycoprotein D antigen of Herpes simplex virus type 2 Choi, Y. S.;T. U. Kim;H. H. Lee;M. H. Cho
  5. Cell v.84 Toxoplasma invasion of mammalian cells is powered by the actin cytoskeleton of the parasite Dobrowolski, J. M.;L. D. Sibley https://doi.org/10.1016/S0092-8674(00)81071-5
  6. Parasitol. Res. v.75 Detection and localization of actin in Toxoplasma gondii Endo, T.;K. Yagita;T. Yasuda;T. Nakamura https://doi.org/10.1007/BF00932708
  7. J. Parasitol. v.84 Actin-dependent motility in Cryptosporidium parvum sporozoites Forney, J. R.;D. K. Vaughan;S. Yang;M. C. Healey https://doi.org/10.2307/3284619
  8. Parasitol. Today v.10 Attachment and invasion of host cells by Toxoplasma gondii Kasper, L. H.;J. R. Mineo https://doi.org/10.1016/0169-4758(94)90026-4
  9. Parasitol. Res. v.87 Nuclear factor-kappa B plays a major role in the regulation of chemokine expression of HeLa cells in response to Toxoplasma gondii infection Kim, J. M.;Y. K. Oh;Y. J. Kim;S. J. Cho;M. H. Ahn;Y. J. Cho https://doi.org/10.1007/s004360100447
  10. Korean J. Parasitol. v.39 Toxoplasma gondii: Ultrastructural localization of specific antigens of intracellular multiplication by monoclonal antibodies Lee, B. Y.;M. H. Ahn;H. C. Kim;D. Y. Min https://doi.org/10.3347/kjp.2001.39.1.67
  11. J. Cell. Science v.108 Invasion of Toxoplasma gondii occurs by active penetration of the host cell Morisaki, J. H.;J. E. Heuser;L. D. Sibley
  12. Exp. Parasitol. v.79 Characterization of extreme apical antigens from Toxoplasma gondii Morrissette, N. S.;V. Bedian;P. Webster;D. S. Roos https://doi.org/10.1006/expr.1994.1106
  13. J. Protozool. v.34 Cytoskeleton of Toxoplasma gondii Nichols, B.;M. L. Chiappino https://doi.org/10.1111/j.1550-7408.1987.tb03162.x
  14. Parasitol. Res. v.87 Improved methods for examination of Toxoplasma gondii cytoskeleton at ultrastructural level Oliveira-Lima, J. G.;J. R. Mineo;A. A. D. Santos;G. L. S. Ferreira;E. A. V. Ferro;C. A. Oliveira https://doi.org/10.1007/PL00008580
  15. Parasitol. v.87 Host cell invasion by apicomplexa: An expression of the parasites contractile system Russell, D. G. https://doi.org/10.1017/S0031182000052562
  16. Infect. Immun. v.20 Effect of cytochalasin on Toxoplasma gondii cell entry Ryning, F. W.;J. S. Remington
  17. J. Microbiol. Biotechnol. v.11 In vitro activities of 2,2'-dipyridyl against Trichomonas vaginalis, Candida albicans, and Gardnerella vaginalis Ryu, J. S.;D. Y. Min;M. C. Kim;N. S. Kim;M. H. Shin
  18. J. Protozool. v.33 Detection of microtubules of the flagellate Trichomonas vaginalis by monoclonal antibodies specific for β-tubulin Schwartzman, J. D.;E. C. Krug https://doi.org/10.1111/j.1550-7408.1986.tb05665.x
  19. J. Protozool. v.32 Detection of the microtubule cytoskeleton of the coccidian Toxoplasma gondii and hemoflagellate Leishmania donovani by monoclonal antibodies specific for β-tubulin Schwartzman, J. D.;E. C. Krug;L. I. Binder;M. R. Rayne https://doi.org/10.1111/j.1550-7408.1985.tb03115.x
  20. Cell Biol. v.96 Induction of an acrosomal process in Toxoplasma gondii: Visualization of actin filaments in a protozoan parasite Shaw, M. K.;L. G. Tilney
  21. J. Cell Science v.113 Microtubules, but not actin filaments, drive daughter cell budding and cell division in Toxoplasma gondii Shaw, M. K.;H. L. Compton;D. S. Ross;L. G. Tilney
  22. Trends. Cell Biol. v.5 Invasion of vertebrate cells by Toxoplasma gondii Sibley, L. D. https://doi.org/10.1016/S0962-8924(00)88964-3
  23. Proc. Natl. Acad. Sci. USA v.76 Electrophoretic transfer of proteins from polyacylamide gels to nitrocellulose sheets: Procedure and some applications Towbin, H.;T. Staehelin;J. Gordon https://doi.org/10.1073/pnas.76.9.4350
  24. Infect. Immun. v.61 Microtubule inhibitors block Cryptosporidium parvum infection of a human enterocyte cell line Wiest, P. M.;J. H. Johnson;T. P. Flanigan
  25. Parasitol. Res. v.75 Immunocytochemical localization of actin in Toxoplasma gondii Yasuda, T.;K. Yagita;T. Nakamura;T. Endo https://doi.org/10.1007/BF00932709
  26. J. Microbiol. Biotechnol. v.10 Encystation of Giardia lamblia by high bile and alkaline pH and its ultrastructural changes during encystation Yong, T. S.;H. W. Yang;K. I. Im;S. J. Park
  27. Korean J. Parasitol. v.38 The effect of microfilament inhibitor on the Cryptosporidium infection in vitro Yu, J. R.;S. D. Choi https://doi.org/10.3347/kjp.2000.38.4.257