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Function of Neospora caninum dense granule protein 7 in innate immunity in mice

  • Immunology and Host-Parasite Interactions - Original Paper
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Abstract

The intracellular parasite Neospora caninum can parasitize all nucleated cells of the host. Dense granule proteins (GRAs) secreted by dense granules are an important material involved in the formation of parasitophorous vacuoles (PVs), which facilitate parasite survival and replication in host cells. Due to the secretory and immune properties of NcGRA7, it is considered to be a promising serodiagnosis marker and an effective neosporosis vaccine candidate. However, the intracellular regulatory mechanisms involved in NcGRA7-induced host responses have rarely been examined. Here, we used the CRISPR/Cas9 genome editing system to obtain a NcGRA7 knockout strain (ΔNcGRA7) and a NcGRA7 complementary strain (iΔNcGRA7) to study their function. We found that ΔNcGRA7 exhibited slower growth in vitro and weakened virulence in mice compared with Nc1 and iΔNcGRA7. All parasite strains can stimulate host immune cells to produce IFN-γ, and the amount of IFN-γ production stimulated by Nc1 was significantly higher than that stimulated by ΔNcGRA7. The transcription levels of the cellular immune factors GBP1, GBP2, IRGa6, and IRGb6 were significantly higher after stimulation with ΔNcGRA7 parasites than after stimulation with Nc1. Furthermore, ΔNcGRA7 infection resulted in greater IRGa6 recruitment to the PVM than Nc1 infection. ΔNcGRA7 parasites were more easily cleared by macrophages than Nc1 parasites. Collectively, these results showed that NcGRA7 plays an important role in regulating the immune factors of mice and the aggregation of IRGa6 at the PVM, which affects the pathogenicity of N. caninum.

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Funding

This study was supported by the National Natural Science Foundation of China (31772730, 31972700).

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Authors and Affiliations

  1. National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, People’s Republic of China

    Fei Wang, Xianmei Wang, Xingju Song, Lei Ma, Jing Yang, Qun Liu & Jing Liu

  2. Key Laboratory of Animal Epidemiology of the Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, People’s Republic of China

    Qun Liu & Jing Liu

Authors
  1. Fei Wang
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  2. Xianmei Wang
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  3. Xingju Song
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  4. Lei Ma
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  5. Jing Yang
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  6. Qun Liu
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  7. Jing Liu
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Contributions

JL conceived the project. FW and LM performed the experiments and drafted the manuscript. JL and QL participated in the design of the study and helped to draft the manuscript. XW, SX, JX, and JY participated in the interpretation of the data. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Jing Liu.

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The authors declare that they have no competing interests.

Ethics approval and consent to participate

The experiments were performed strictly according to the recommendations of the Guide for the Care and Use of Laboratory Animals of the Ministry of Science and Technology of China. All experimental animal procedures and protocols were approved by the Institutional Animal Care and Use Committee of China Agricultural University.

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Section Editor: Xing-Quan ZHU

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Wang, F., Wang, X., Song, X. et al. Function of Neospora caninum dense granule protein 7 in innate immunity in mice. Parasitol Res 120, 197–207 (2021). https://doi.org/10.1007/s00436-020-06961-4

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  • DOI: https://doi.org/10.1007/s00436-020-06961-4

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