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Analysis of the virulence determination mechanisms in a local Toxoplasma strain (T.gHB1) isolated from central China

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Abstract

Several rhoptry proteins (ROPs) have been confirmed to be critical virulence factors of Toxoplasma gondii strains from North America and Europe. The two active kinases ROP17 and ROP18, and pseudokinase ROP5 were thought to be the key determinants of parasites’ virulence in laboratory mice. Given the genetic diversity of Toxoplasma strains from different geographical regions, the virulence determinants in other strains, particularly the ones that are phylogenetically distant to the North American and European strains, are yet to be elucidated. In this study, we sought to examine the contribution of three known virulence factors to the virulence of a type I strain (T.gHB1) isolated from Central China. We deleted ROP17 and ROP18 individually, as well as in combination with GRA7 by the CRISPR-Cas9 system in this local isolate. Subsequent virulence tests in mice indicated that deletion of GRA7, ROP17, or ROP18 in T.gHB1showed similar attenuation in mice as the type I RH strain lacking the corresponding proteins. However, in contrast to the reported double knockouts in RH, double deletions of GRA7 plus ROP17 or GRA7 plus ROP18 in T.gHB1 did not show significant further virulence attenuation compared to the ROP17 or ROP18 single knockouts. These results indicated that GRA7, ROP18 and ROP17 may play different roles in virulence determination in genetically diverse strains of Toxoplasma.

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Acknowledgments

The authors thanked Dr. Aditi Alaganan (Pasteur Institute, Paris, France) for her critical reading and valuable suggestions for the improvement of this manuscript. This work was supported by the National Natural Science Foundation of China (Grants No. 31372429 and No. 31572510), National Key Basic Research Program (973program) of China (Grant No. 2015CB150302), and Project 2662015PY048 from the Fundamental Research Funds for the Central Universities in China.

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Authors and Affiliations

  1. State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, Hubei, China

    Weichao Zhang, Longjiao Li, Ningbo Xia, Yanqin Zhou, Rui Fang, Lan He, Min Hu, Bang Shen & Junlong Zhao

  2. Key Laboratory for development of veterinary diagnostic products, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, Hubei, China

    Weichao Zhang, Longjiao Li, Ningbo Xia, Yanqin Zhou, Rui Fang, Lan He, Min Hu, Bang Shen & Junlong Zhao

  3. Hubei Cooperative Innovation Center for Sustainable Pig Production, Wuhan, 430070, Hubei, China

    Junlong Zhao

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  1. Weichao Zhang
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  9. Junlong Zhao
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Correspondence to Bang Shen or Junlong Zhao.

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Human and animals rights informed consent

All the ICR mice and rabbits used in all experiments were purchased from Laboratory Animals Research Centre of Hubei Province (permit number SCXK (E)-2015-0018) and raised under the standard conditions according to the Regulations for the Administration of Affairs Concerning Experimental Animals. The animal experiments were approved by the ethical committee of Huazhong Agricultural University according to the Regulations of the Care and Use of Laboratory Animals in China.

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Zhang, W., Li, L., Xia, N. et al. Analysis of the virulence determination mechanisms in a local Toxoplasma strain (T.gHB1) isolated from central China. Parasitol Res 115, 3807–3815 (2016). https://doi.org/10.1007/s00436-016-5141-z

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