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Toxoplasma gondii ROP18: potential to manipulate host cell mitochondrial apoptosis

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Abstract

Toxoplasma gondii is an obligate intracellular parasite that may manipulate host cell mitochondrial apoptosis pathways. In our experiment, 293T cells were transfected with the p3×FLAG-CMV-Myc-ROP18 vector and expressed the ROP18-Myc fusion protein. Cell apoptosis was induced by 0.5 μg/mL actinomycin D (ActD) and was detected by Annexin V-FITC/PI assay. The cell mitochondrial membrane potential was determined by JC-1. Cytochrome c (Cyto-c) from mitochondria and the cytoplasm was measured by Western blot. The Bcl-2 and Bax coding gene expression levels were detected by real-time PCR. We found, in vitro, that T. gondii ROP18 significantly suppressed 293T cell apoptosis induced by ActD and maintained mitochondrial membrane potential and integrity, thereby preventing the release of Cyto-c from mitochondria into the cytoplasm. The ratio of Bcl-2/Bax in ROP18-overexpressing cells was significantly higher than that of the negative control. Therefore, we speculate that ROP18 could suppress host cell apoptosis via the mitochondrial apoptosis pathway in vitro.

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Acknowledgments

This study received financial support from the National Natural Science Foundation of China (grant number 81301453), the Laboratory of Parasite and Vector Biology of China, MOPH (grant number WSBKTKT201302), the China Postdoctoral Science Special Foundation(grant number 2015T80518), the China Postdoctoral Science Foundation (grant number 2014M561598), Jiangsu Postdoctoral Science Foundation (grant number 1402171C), and the Senior Talent Studying Initial Funding of Jiangsu University(grant number 13JDG023, 13JDG127).

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    Authors and Affiliations

    1. National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Key Laboratory of Parasite and Vector Biology, WHO Collaborating Center for Tropical Diseases, National Center of International Research on Tropical Diseases, Shanghai, 200025, China

      Liang Wu & Jianping Cao

    2. School of Medicine, Jiangsu University, Zhenjiang, 212013, China

      Liang Wu, Xiao Wang, Yunhui Li, Yuan Liu, Danhua Su, Tao Fu, Fei Guo, Liangping Gu, Xugan Jiang & Shengxia Chen

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    1. Liang Wu
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    2. Xiao Wang
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    3. Yunhui Li
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    4. Yuan Liu
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    Correspondence to Shengxia Chen or Jianping Cao.

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    Liang Wu and Xiao Wang contributed equally to this work.

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    Wu, L., Wang, X., Li, Y. et al. Toxoplasma gondii ROP18: potential to manipulate host cell mitochondrial apoptosis. Parasitol Res 115, 2415–2422 (2016). https://doi.org/10.1007/s00436-016-4993-6

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    • DOI: https://doi.org/10.1007/s00436-016-4993-6

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