Abstract
Avian reovirus (ARV)-induced apoptosis contributes to the pathogenesis of reovirus in infected chickens. However, methods for effectively reducing ARV-triggered apoptosis remain to be explored. Here, we show that pretreatment with chloroquine (CQ) or E64d plus pepstatin A decreases ARV-mediated apoptosis in chicken DF-1 cells. By acting as autophagy inhibitors, CQ and E64d plus pepstatin A increase microtubule-associated protein 1 light chain 3-II (LC3II) accumulation in ARV-infected cells, which results in decreased ARV protein synthesis and virus yield and thereby contributes to the reduction of apoptosis. Furthermore, ARV-mediated apoptosis in the bursa, heart and intestines of chicken embryos is attenuated by CQ and E64d plus pepstatin A treatment. Importantly, treatment with these autophagy inhibitors increases the survival of infected chicken embryos. Together, our data suggest that pharmacological inhibition of autophagy might represent a novel strategy for reducing ARV-mediated apoptosis.
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References
Amaravadi RK, Yu D, Lum JJ, Bui T, Christophorou MA, Evan GI, Thomas-Tikhonenko A, Thompson CB (2007) Autophagy inhibition enhances therapy-induced apoptosis in a Myc-induced model of lymphoma. J Clin Investig 117:326–336
Benavente J, Martinez-Costas J et al (2007) Avian reovirus: structure and biology. Virus Res 123:105–119
Bian J, Wang K, Kong X, Liu H, Chen F, Hu M, Zhang X, Jiao X, Ge B, Wu Y, Meng S (2011) Caspase- and p38-MAPK-dependent induction of apoptosis in A549 lung cancer cells by Newcastle disease virus. Arch Virol 156:1335–1344
Chen WT, Wu YL, Chen T, Cheng CS, Chan HL, Chou HC, Chen YW, Yin HS (2014) Proteomics analysis of the DF-1 chicken fibroblasts infected with avian reovirus strain S1133. PloS One 9:e92154
Chen YT, Lin CH, Ji WT, Li SK, Liu HJ (2008) Proteasome inhibition reduces avian reovirus replication and apoptosis induction in cultured cells. J Virol Methods 151:95–100
Chi PI, Huang WR, Lai IH, Cheng CY, Liu HJ (2013) The p17 nonstructural protein of avian reovirus triggers autophagy enhancing virus replication via activation of phosphatase and tensin deleted on chromosome 10 (PTEN) and AMP-activated protein kinase (AMPK), as well as dsRNA-dependent protein kinase (PKR)/eIF2alpha signaling pathways. J Biol Chem 288:3571–3584
Chulu JL, Lee LH, Lee YC, Liao SH, Lin FL, Shih WL, Liu HJ (2007) Apoptosis induction by avian reovirus through p53 and mitochondria-mediated pathway. Biochem Biophys Res Commun 356:529–535
Crawford SE, Hyser JM, Utama B, Estes MK (2012) Autophagy hijacked through viroporin-activated calcium/calmodulin-dependent kinase kinase-beta signaling is required for rotavirus replication. Proc Natl Acad Sci USA 109:E3405–E3413
Ji WT, Lee LH, Lin FL, Wang L, Liu HJ (2009) AMP-activated protein kinase facilitates avian reovirus to induce mitogen-activated protein kinase (MAPK) p38 and MAPK kinase 3/6 signalling that is beneficial for virus replication. J Gen Virol 90:3002–3009
Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T (2000) LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. The EMBO journal 19:5720–5728
Klionsky DJ, Abeliovich H (2008) Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy 4:151–175
Lin PY, Lee JW, Liao MH, Hsu HY, Chiu SJ, Liu HJ, Shih WL (2009) Modulation of p53 by mitogen-activated protein kinase pathways and protein kinase C delta during avian reovirus S1133-induced apoptosis. Virology 385:323–334
Lin PY, Liu HJ, Liao MH, Chang CD, Chang CI, Cheng HL, Lee JW, Shih WL (2010) Activation of PI 3-kinase/Akt/NF-kappaB and Stat3 signaling by avian reovirus S1133 in the early stages of infection results in an inflammatory response and delayed apoptosis. Virology 400:104–114
Meng S, Jiang K, Zhang X, Zhang M, Zhou Z, Hu M, Yang R, Sun C, Wu Y (2012) Avian reovirus triggers autophagy in primary chicken fibroblast cells and Vero cells to promote virus production. Arch Virol 157:661–668
Ni HM, Bockus A, Wozniak AL, Jones K, Weinman S, Yin XM, Ding WX (2011) Dissecting the dynamic turnover of GFP-LC3 in the autolysosome. Autophagy 7:188–204
Ping-Yuan L, Hung-Jen L, Meng-Jiun L, Feng-Ling Y, Hsue-Yin H, Jeng-Woei L, Wen-Ling S (2006) Avian reovirus activates a novel proapoptotic signal by linking Src to p53. Apoptosis 11:2179–2193
Roessler DE, Rosenberger JK (1989) In vitro and in vivo characterization of avian reoviruses. III. Host factors affecting virulence and persistence. Avian Dis 33:555–565
Shih WL, Hsu HW, Liao MH, Lee LH, Liu HJ (2004) Avian reovirus sigmaC protein induces apoptosis in cultured cells. Virology 321:65–74
Talloczy Z, Jiang W, Virgin HWt, Leib DA, Scheuner D, Kaufman RJ, Eskelinen EL, Levine B (2002) Regulation of starvation- and virus-induced autophagy by the eIF2alpha kinase signaling pathway. Proc Natl Acad Sci USA 99:190–195
Acknowledgments
We thank Dr. Songshu Meng at Dalian Medical University Cancer Center for suggestions. This work was financially supported by National Natural Science Foundation of China (31272576), the China Agriculture Research System (CARS-41-K08) and the Priority Academic Program Development of Jiangsu Higher Education Institutions.
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Duan, S., Cheng, J., Li, C. et al. Autophagy inhibitors reduce avian-reovirus-mediated apoptosis in cultured cells and in chicken embryos. Arch Virol 160, 1679–1685 (2015). https://doi.org/10.1007/s00705-015-2415-1
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DOI: https://doi.org/10.1007/s00705-015-2415-1