Abstract
Enterovirus D68 (EVD68) is an emerging pathogen that recently caused a large worldwide outbreak of severe respiratory disease in children. However, the relationship between EVD68 and host cells remains unclear. Caspases are involved in cell death, immune response, and even viral production. We found that caspase-3 was activated during EVD68 replication to induce apoptosis. Caspase-3 inhibitor (Z-DEVD-FMK) inhibited viral production, protected host cells from the cytopathic effects of EVD68 infection, and prevented EVD68 from regulating the host cell cycle at G0/G1. Meanwhile, caspase-3 activator (PAC-1) increased EVD68 production. EVD68 infection therefore activates caspase-3 for virus production. This knowledge provides a potential direction for the prevention and treatment of disease related to EVD68.
Article PDF
Similar content being viewed by others
References
Carballo, C.M., Erro, M.G., Sordelli, N., Vazquez, G., Mistchenko, A.S., Cejas, C., Rodriguez, M., Cisterna, D.M., Freire, M.C., Contrini, M.M., et al. 2019. Acute flaccid myelitis associated with enterovirus D68 in children, argentina, 2016. Emerg. Infect. Dis. 25, 573–576.
Chang, S.C., Lin, J.Y., Lo, L.Y., Li, M.L., and Shih, S.R. 2004. Diverse apoptotic pathways in enterovirus 71-infected cells. J. Neurovirol. 10, 338–349.
Esposito, S., Zampiero, A., Ruggiero, L., Madini, B., Niesters, H., and Principi, N. 2015. Enterovirus D68-associated community-acquired pneumonia in children living in Milan, Italy. J. Clin. Virol. 68, 94–96.
Farrell, J.J., Ikladios, O., Wylie, K.M., O’Rourke, L.M., Lowery, K.S., Cromwell, J.S., Wylie, T.N., Melendez, E.L.V., Makhoul, Y., Sampath, R., et al. 2015. Enterovirus D68-associated acute respiratory distress syndrome in adult, United States, 2014. Emerg. Infect. Dis. 21, 914–916.
Gay, R.T., Belisle, S., Beck, M.A., and Meydani, S.N. 2006. An aged host promotes the evolution of avirulent coxsackievirus into a virulent strain. Proc. Natl. Acad. Sci. USA 103, 13825–13830.
Hardwick, J.M. 1998. Viral interference with apoptosis. Semin. Cell Dev. Biol. 9, 339–349.
Henderson, S., Huen, D., Rowe, M., Dawson, C., Johnson, G., and Rickinson, A. 1993. Epstein-Barr virus-coded BHRF1 protein, a viral homologue of Bcl-2, protects human B cells from programmed cell death. Proc. Natl. Acad. Sci. USA 90, 8479–8483.
Hengartner, M.O. 2000. The biochemistry of apoptosis. Nature 407, 770–776.
Holmes, K.V. 1975. Scanning electron microscopic studies of virus-infected cells. I. Cytopathic effects and maturation of vesicular stomatitis virus in L2 cells. J. Virol. 15, 355–362.
Kamau, E., Harvala, H., Blomqvist, S., Nguyen, D., Horby, P., Pebody, R., and Simmonds, P. 2019. Increase in enterovirus D68 infections in young children, United Kingdom, 2006–2016. Emerg. Infect. Dis. 25, 1200–1203.
Li, Z., Yu, J., Liu, L., Wei, Z., Ehrlich, E.S., Liu, G., Li, J., Liu, X., Wang, H., Yu, X., et al 2014. Coxsackievirus A16 infection induces neural cell and non-neural cell apoptosis in vitro. PLoS ONE 9, e111174.
Lu, J.R., Lu, W.W., Lai, J.Z., Tsai, F.L., Wu, S.H., Lin, C.W., and Kung, S.H. 2013. Calcium flux and calpain-mediated activation of the apoptosis-inducing factor contribute to enterovirus 71-induced apoptosis. J. Gen. Virol. 94, 1477–1485.
Martin, U., Jarasch, N., Nestler, M., Rassmann, A., Munder, T., Seitz, S., Zell, R., Wutzler, P., and Henke, A. 2007. Antiviral effects of pan-caspase inhibitors on the replication of coxsackievirus B3. Apoptosis 12, 525–533.
Messacar, K. and Tyler, K.L. 2019. Enterovirus D68-associated acute flaccid myelitis: rising to the clinical and research challenges. JAMA 321, 831–832.
Oberste, M.S., Maher, K., Kilpatrick, D.R., Flemister, M.R., Brown, B.A., and Pallansch, M.A. 1999a. Typing of human enteroviruses by partial sequencing of VP1. J. Clin. Microbiol. 37, 1288–1293.
Oberste, M.S., Maher, K., Kilpatrick, D.R., and Pallansch, M.A. 1999b. Molecular evolution of the human enteroviruses: correlation of serotype with VP1 sequence and application to picornavirus classification. J. Virol. 73, 1941–1948.
Rao, L., Modha, D., and White, E. 1997. The E1B 19K protein associates with lamins in vivo and its proper localization is required for inhibition of apoptosis. Oncogene 15, 1587–1597.
Reed, L.J. and Muench, H. 1938. A simple method of estimating fifty percent endpoints. Am. J. Epidemiol. 27, 493–497.
Reiche, J., Böttcher, S., Diedrich, S., Buchholz, U., Buda, S., Haas, W., Schweiger, B., and Wolff, T. 2015. Low-level circulation of enterovirus D68-associated acute respiratory infections, Germany, 2014. Emerg. Infect. Dis. 21, 837–841.
Richard, A. and Tulasne, D. 2012. Caspase cleavage of viral proteins, another way for viruses to make the best of apoptosis. Cell Death Dis. 3, e277.
Salvesen, G.S. and Dixit, V.M. 1997. Caspases: intracellular signaling by proteolysis. Cell 91, 443–446.
Shi, W., Li, X., Hou, X., Peng, H., Jiang, Q., Shi, M., Ji, Y., Liu, X., and Liu, J. 2012. Differential apoptosis gene expressions of rhabdomyosarcoma cells in response to enterovirus 71 infection. BMC Infect. Dis. 12, 327.
Smura, T., Ylipaasto, P., Klemola, P., Kaijalainen, S., Kyllönen, L., Sordi, V., Piemonti, L., and Roivainen, M. 2010. Cellular tropism of human enterovirus D species serotypes EV-94, EV-70, and EV-68 in vitro: implications for pathogenesis. J. Med. Virol. 82, 1940–1949.
Song, F., Yu, X., Zhong, T., Wang, Z., Meng, X., Li, Z., Zhang, S., Huo, W., Liu, X., Zhang, Y., et al. 2018. Caspase-3 inhibition attenuates the cytopathic effects of EV71 infection. Front. Microbiol. 9, 817.
Villa, P., Kaufmann, S.H., and Earnshaw, W.C. 1997. Caspases and caspase inhibitors. Trends Biochem. Sci. 22, 388–393.
Wang, Z., Wang, Y., Wang, S., Meng, X., Song, F., Huo, W., Zhang, S., Chang, J., Li, J., Zheng, B., et al. 2018. Coxsackievirus A6 induces cell cycle arrest in G0/G1 phase for viral production. Front. Cell. Infect. Microbiol. 8, 279.
Wang, Z., Zhong, T., Wang, Y., Song, F., Yu, X., Xing, L., Zhang, W., Yu, J., Hua, S., and Yu, X. 2017. Human enterovirus 68 interferes with the host cell cycle to facilitate viral production. Front. Cell. Infect. Microbiol. 7, 29.
Wang, C., Zhou, R., Zhang, Z., Jin, Y., Cardona, C.J., and Xing, Z. 2015. Intrinsic apoptosis and proinflammatory cytokines regulated in human astrocytes infected with enterovirus 71. J. Gen. Virol. 96, 3010–3022.
Wurzer, W.J., Planz, O., Ehrhardt, C., Giner, M., Silberzahn, T., Pleschka, S., and Ludwig, S. 2003. Caspase 3 activation is essential for efficient influenza virus propagation. EMBO J. 22, 2717–2728.
Yu, J., Cui, Q., Jiang, Y., Yang, W., Tashiro, S., Onodera, S., and Ikejima, T. 2007. Pseudolaric acid B induces apoptosis, senescence, and mitotic arrest in human breast cancer MCF-7. Acta Pharmacol. Sin. 28, 1975–1983.
Yu, J., Wang, Z., Ren, P., Zhong, T., Wang, Y., Song, F., Hou, J., Yu, X., and Hua, S. 2017. Pseudolaric acid B inhibits the secretion of hepatitis B virus. Oncol. Rep. 37, 519–525.
Yu, J., Zhang, L., Ren, P., Zhong, T., Li, Z., Wang, Z., Li, J., Liu, X., Zhao, K., Zhang, W., et al 2015. Enterovirus 71 mediates cell cycle arrest in S phase through non-structural protein 3D. Cell Cycle 14, 425–436.
Zhang, T., Ren, L., Luo, M., Li, A., Gong, C., Chen, M., Yu, X., Wu, J., Deng, Y., and Huang, F. 2015. Enterovirus D68-associated severe pneumonia, China, 2014. Emerg. Infect. Dis. 21, 916–918.
Zhang, S., Yu, X., Meng, X., Huo, W., Su, Y., Liu, J., Liu, Y., Zhang, J., Wang, S., and Yu, J. 2020. Coxsackievirus A6 induces necroptosis for viral production. Front. Microbiol. 11, 42.
Acknowledgments
This work was supported by the National Natural Science Foundation of China (81871634 and 81301416), Postdoctoral Science Foundation of China (2014M561302 and 2015-T80299), Norman Bethune Program of Jilin University (2015–202), the Jilin Provincial Science and Technology Department (20140204004YY, 20160414025GH and 20190304064YY), and the Department of Human Resources and Social Security of Jilin Province (2016014).
Author information
Authors and Affiliations
Corresponding author
Additional information
Conflict of Interest
The authors declare no conflict of interest.
Rights and permissions
About this article
Cite this article
Huo, W., Yu, J., Liu, C. et al. Caspase-3 inhibitor inhibits enterovirus D68 production. J Microbiol. 58, 812–820 (2020). https://doi.org/10.1007/s12275-020-0241-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12275-020-0241-y