Abstract
Heat-shock protein 70 (HSP70) is highly expressed in Toxoplasma gondii-infected cells. However, the role of this protein is not well understood, especially during apoptosis. This study addresses the mechanism behind the antiapoptotic chaperone activity of HSP70 in Toxoplasma-infected host cells using a human macrophage cell line, THP-1 by Western blot, DNA fragmentation assay, immunoprecipitation, and a caspase-3/7 activity assay based on cleavage of the colorimetric substrate DEVD-pNA. Apoptosis induced by arsenic trioxide (As2O3) was inhibited in T. gondii-infected THP-1 cells, but not in uninfected cells. Without As2O3 induction of apoptosis, T. gondii infection caused increased expression of Bcl-2 and HSP70, but not caspase-3. However, active form caspase-3 levels were lower in As2O3-treated infected cells as compared with As2O3-treated uninfected cells. Bcl-2 expression in As2O3-treated infected cells was similar to that in cells infected with T. gondii. Translocation of apoptosis-inducing factor (AIF) and release of cytochrome c from mitochondria were inhibited in As2O3-treated infected cells as compared with As2O3-treated uninfected cells. Increased parasite loads in Toxoplasma-infected macrophages caused higher HSP70 and Bcl-2 expression in whole-cell extracts and fractionated components, respectively. However, expression of AIF and cytochrome c was unaffected. Toxoplasma dose-dependently inhibited caspase-3 activation, thus revealing an anti-apoptotic parasite activity on cytochrome c-mediated caspase activation in subcellular components. In addition, immunoprecipitation analysis suggested that HSP70 is capable of binding to the pro-apoptotic factors AIF and Apaf-1, but not to cytochrome c or procaspase-9. Taken together, these data demonstrate that T. gondii infection inhibits mitochondrial apoptosis through overproduction of anti-apoptotic Bcl-2 as well as HSP70, which are increased parasite loads dependently.
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Acknowledgements
This work was supported by National Research Foundation of Korea Grant funded by the Korean Government (KRF-2008-314-E00070-7644), and by a Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of Education, Science, and Technology (R13-2007-020-00000-0).
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First co-authors: Il-Young Hwang, Juan Hua Quan, and Myoung-Hee Ahn
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Hwang, IY., Quan, J.H., Ahn, MH. et al. Toxoplasma gondii infection inhibits the mitochondrial apoptosis through induction of Bcl-2 and HSP70. Parasitol Res 107, 1313–1321 (2010). https://doi.org/10.1007/s00436-010-1999-3
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DOI: https://doi.org/10.1007/s00436-010-1999-3