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Rotavirus NSP486–175 interacts with H9c2(2-1) cells in vitro, elevates intracellular Ca2+ levels and can become cytotoxic: a possible mechanism for extra-intestinal pathogenesis

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Abstract

Rotavirus (RV) is the predominant cause of infantile gastroenteritis with multiple pathogenic factors, among which enterotoxin NSP4 is the most significant factor. NSP4 has been shown to induce elevation of the intracellular calcium concentration, alteration of the cytoskeleton organization, and cytopathic effect among other processes. However, increasing evidence suggests that RVs can escape from the gastrointestinal tract and invade other organs and tissues to cause extra-intestinal diseases. In this study, we investigated whether NSP4 has a pathogenic effect on extra-intestinal cells and examined possible molecular mechanisms in vitro. Our results showed that NSP486–175 has important functions in increasing intracellular Ca2+ concentration, altering actin cytoskeleton organization and inducing cellular damage in H9c2(2-1) cells. Blockade of the integrin α2 receptor using a specific antibody attenuated the increase of intracellular Ca2+ concentration and alleviated the observed cytopathic effects, suggesting that integrin α2 may be a receptor for NSP486–175. Collectively, these results indicate that extracellular NSP486–175 can induce elevation of the intracellular Ca2+ concentration, cause cytotoxic changes, and disrupt the actin cytoskeleton in H9c2(2-1) cells, which may constitute a possible mechanism for RV extra-intestinal pathogenesis.

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Abbreviations

RVs:

Rotaviruses

NSP4:

Non-structural protein 4

NSP486–175 :

Non-structural protein four fragment spanning amino acids 86–175

[Ca2+]i:

Cytoplasmic calcium concentration

LDH:

Lactate dehydrogenase

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Acknowledgements

This work was supported by the Natural Science Foundation of Zhejiang Province, China (LY14H190006).

Author contributions

XX and XL designed the research. YH and CL helped with the experiments, data analysis, and discussion. XX and XL wrote the manuscript.

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Authors and Affiliations

  1. Department of Clinical Laboratory, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325027, Zhejiang, People’s Republic of China

    Xiaoshun Xiong, Yinyin Hu, Caixia Liu & Xiangyang Li

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  1. Xiaoshun Xiong
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  2. Yinyin Hu
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Correspondence to Xiangyang Li.

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Xiong, X., Hu, Y., Liu, C. et al. Rotavirus NSP486–175 interacts with H9c2(2-1) cells in vitro, elevates intracellular Ca2+ levels and can become cytotoxic: a possible mechanism for extra-intestinal pathogenesis. Virus Genes 53, 179–189 (2017). https://doi.org/10.1007/s11262-016-1419-6

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