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Volume 96, Issue 10

Intrinsic apoptosis and proinflammatory cytokines regulated in human astrocytes infected with enterovirus 71 Free

Abstract

Enterovirus 71 (EV71) has emerged as a clinically important neurotropic virus following poliovirus eradication. However, the mechanism of EV71-induced neurological manifestation remains largely unclear. In this study, we showed that human astrocytes were susceptible to EV71 and viral RNA was first detected at 12 h post-infection (p.i.), whilst viral proteins were detected at 36 h p.i. EV71-infected astrocytes underwent apoptosis, in which cytochrome was released from mitochondria to the cytosol and caspase-9 was activated. Interestingly, caspase-2 and -8 were not cleaved or activated during the infection, whilst a selective inhibitor of caspase-9, Z-LEHD-FMK, blocked the cleavage of caspase-3 and -7, indicating that only the mitochondria-mediated intrinsic apoptotic pathway was activated in EV71-infected astrocytes. EV71 infection also induced proinflammatory cytokines, including IL-6, IL-8, CCL5 and IFN-γ-inducible protein (IP)-10 in astrocytes, which may play a critical role in EV71-induced neuroinflammation and neurological complications. By using inhibitors of mitogen-activated protein kinases (MAPKs), we demonstrated that the induction of the cytokines was mainly regulated by the MAPK p38 signalling pathway as a significant reduction of the cytokines was observed when treated with p38 inhibitors. This study demonstrated that human astrocytes were susceptible to EV71, and the infection led to intrinsic apoptosis and induction of p38-regulated proinflammatory cytokines. These findings further our understanding of the neuropathogenesis in severe cases of EV71 infection.

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© 2015 The Authors
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2015-10-01
2024-04-27
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References

  1. Aschner M. 1998; Immune and inflammatory responses in the CNS: modulation by astrocytes. Toxicol Lett 102-103:283–287 [View Article][PubMed]
     [Google Scholar]
  2. Baker A.B. 1957; Poliomyelitis. 16. A study of pulmonary edema. Neurology 7:743–751 [View Article][PubMed]
     [Google Scholar]
  3. Boatright K.M., Salvesen G.S. 2003; Mechanisms of caspase activation. Curr Opin Cell Biol 15:725–731 [View Article][PubMed]
     [Google Scholar]
  4. Camelo S., Lafage M., Lafon M. 2000; Absence of the p55 Kd TNF-alpha receptor promotes survival in rabies virus acute encephalitis. J Neurovirol 6:507–518 [View Article][PubMed]
     [Google Scholar]
  5. Chang S.-C., Lin J.-Y., Lo L.Y.-C., Li M.-L., Shih S.-R. 2004; Diverse apoptotic pathways in enterovirus 71-infected cells. J Neurovirol 10:338–349 [View Article][PubMed]
     [Google Scholar]
  6. Chaparro-Huerta V., Rivera-Cervantes M.C., Torres-Mendoza B.M., Beas-Zárate C. 2002; Neuronal death and tumor necrosis factor-alpha response to glutamate-induced excitotoxicity in the cerebral cortex of neonatal rats. Neurosci Lett 333:95–98 [View Article][PubMed]
     [Google Scholar]
  7. Chen L.C., Shyu H.W., Chen S.H., Lei H.Y., Yu C.K., Yeh T.M. 2006; Enterovirus 71 infection induces Fas ligand expression and apoptosis of Jurkat cells. J Med Virol 78:780–786 [View Article][PubMed]
     [Google Scholar]
  8. Chen C.S., Yao Y.C., Lin S.C., Lee Y.P., Wang Y.F., Wang J.R., Liu C.C., Lei H.Y., Yu C.K. 2007a; Retrograde axonal transport: a major transmission route of enterovirus 71 in mice. J Virol 81:8996–9003 [View Article][PubMed]
     [Google Scholar]
  9. Chen T.C., Lai Y.K., Yu C.K., Juang J.L. 2007b; Enterovirus 71 triggering of neuronal apoptosis through activation of Abl-Cdk5 signalling. Cell Microbiol 9:2676–2688 [View Article][PubMed]
     [Google Scholar]
  10. de Vries H.E., Blom-Roosemalen M.C.M., van Oosten M., de Boer A.G., van Berkel T.J., Breimer D.D., Kuiper J. 1996; The influence of cytokines on the integrity of the blood-brain barrier in vitro . J Neuroimmunol 64:37–43 [View Article][PubMed]
     [Google Scholar]
  11. Dong Y., Benveniste E.N. 2001; Immune function of astrocytes. Glia 36:180–190 [View Article][PubMed]
     [Google Scholar]
  12. Farina C., Aloisi F., Meinl E. 2007; Astrocytes are active players in cerebral innate immunity. Trends Immunol 28:138–145 [View Article][PubMed]
     [Google Scholar]
  13. Fiala M., Looney D.J., Stins M., Way D.D., Zhang L., Gan X., Chiappelli F., Schweitzer E.S., Shapshak P., other authors. 1997; TNF-alpha opens a paracellular route for HIV-1 invasion across the blood-brain barrier. Mol Med 3:553–564
     [Google Scholar]
  14. Franklin J.L. 2011; Redox regulation of the intrinsic pathway in neuronal apoptosis. Antioxid Redox Signal 14:1437–1448 [View Article][PubMed]
     [Google Scholar]
  15. Guo Y., Srinivasula S.M., Druilhe A., Fernandes-Alnemri T., Alnemri E.S. 2002; Caspase-2 induces apoptosis by releasing proapoptotic proteins from mitochondria. J Biol Chem 277:13430–13437 [View Article][PubMed]
     [Google Scholar]
  16. Haolong C., Du N., Hongchao T., Yang Y., Wei Z., Hua Z., Wenliang Z., Lei S., Po T. 2013; Enterovirus 71 VP1 activates calmodulin-dependent protein kinase II and results in the rearrangement of vimentin in human astrocyte cells. PLoS One 8:e73900 [View Article][PubMed]
     [Google Scholar]
  17. Ho M. 2000; Enterovirus 71: the virus, its infections and outbreaks. J Microbiol Immunol Infect 33:205–216
     [Google Scholar]
  18. Hsu Y.T., Wolter K.G., Youle R.J. 1997; Cytosol-to-membrane redistribution of Bax and Bcl-XL during apoptosis. Proc Natl Acad Sci U S A 94:3668–3672 [View Article][PubMed]
     [Google Scholar]
  19. Hsueh C., Jung S.M., Shih S.R., Kuo T.T., Shieh W.J., Zaki S., Lin T.Y., Chang L.Y., Ning H.C., Yen D.C. 2000; Acute encephalomyelitis during an outbreak of enterovirus type 71 infection in Taiwan: report of an autopsy case with pathologic, immunofluorescence, and molecular studies. Mod Pathol 13:1200–1205 [View Article][PubMed]
     [Google Scholar]
  20. Jiao X.Y., Guo L., Huang D.Y., Chang X.L., Qiu Q.C. 2014; Distribution of EV71 receptors SCARB2 and PSGL-1 in human tissues. Virus Res 190:40–52 [View Article][PubMed]
     [Google Scholar]
  21. Kärber G. 1931; [A contribution to the collective treatment of a pharmacological experimental series]. Naunyn Schmiedebergs Arch Exp Pathol Pharmakol 162:480–483 (in German) [CrossRef]
     [Google Scholar]
  22. Khong W.X., Foo D.G., Trasti S.L., Tan E.L., Alonso S. 2011; Sustained High Levels of Interleukin-6 Contribute to the Pathogenesis of Enterovirus 71 in a Neonate Mouse Model. J Virol 85:3067–3076 [CrossRef]
     [Google Scholar]
  23. Kimura Y. 2006; [Cytokines and chemokines induced by influenza virus infection]. Nihon Rinsho 64:1822–1827 (in Japanese)
    [Google Scholar]
  24. Kuo R.L., Kung S.H., Hsu Y.Y., Liu W.T. 2002; Infection with enterovirus 71 or expression of its 2A protease induces apoptotic cell death. J Gen Virol 83:1367–1376 [CrossRef]
     [Google Scholar]
  25. Li M.L., Hsu T.A., Chen T.C., Chang S.C., Lee J.C., Chen C.C., Stollar V., Shih S.R. 2002; The 3C protease activity of enterovirus 71 induces human neural cell apoptosis. Virology 293:386–395 [View Article][PubMed]
     [Google Scholar]
  26. Lin T.Y., Chang L.Y., Hsia S.H., Huang Y.C., Chiu C.H., Hsueh C., Shih S.R., Liu C.C., Wu M.H. 2002a; The 1998 enterovirus 71 outbreak in Taiwan: pathogenesis and management. Clin Infect Dis 34 Suppl 2:S52–S57 [View Article][PubMed]
     [Google Scholar]
  27. Lin T.Y., Chang L.Y., Huang Y.C., Hsu K.H., Chiu C.H., Yang K.D. 2002b; Different proinflammatory reactions in fatal and non-fatal enterovirus 71 infections: implications for early recognition and therapy. Acta Paediatr 91:632–635 [View Article][PubMed]
     [Google Scholar]
  28. Lin T.Y., Hsia S.H., Huang Y.C., Wu C.T., Chang L.Y. 2003; Proinflammatory cytokine reactions in enterovirus 71 infections of the central nervous system. Clin Infect Dis 36:269–274 [View Article][PubMed]
     [Google Scholar]
  29. Lucas S.M., Rothwell N.J., Gibson R.M. 2006; The role of inflammation in CNS injury and disease. Br J Pharmacol 147:(Suppl 1)S232–S240 [View Article][PubMed]
     [Google Scholar]
  30. Lum L.C., Wong K.T., Lam S.K., Chua K.B., Goh A.Y., Lim W.L., Ong B.B., Paul G., AbuBakar S., Lambert M. 1998; Fatal enterovirus 71 encephalomyelitis. J Pediatr 133:795–798 [View Article][PubMed]
     [Google Scholar]
  31. Mayhan W.G. 2002; Cellular mechanisms by which tumor necrosis factor-alpha produces disruption of the blood-brain barrier. Brain Res 927:144–152 [CrossRef]
     [Google Scholar]
  32. Ooi M.H., Wong S.C., Mohan A., Podin Y., Perera D., Clear D., del Sel S., Chieng C.H., Tio P.H., other authors. 2009; Identification and validation of clinical predictors for the risk of neurological involvement in children with hand, foot, and mouth disease in Sarawak. BMC Infect Dis 9:3 [View Article][PubMed]
     [Google Scholar]
  33. Reed L.J., Muench H. 1938; A simple method of estimating fifty per cent endpoints. Am J Epidemiol 27:493–497
     [Google Scholar]
  34. Rhoades R.E., Tabor-Godwin J.M., Tsueng G., Feuer R. 2011; Enterovirus infections of the central nervous system. Virology 411:288–305 [View Article][PubMed]
     [Google Scholar]
  35. Shieh W.J., Jung S.M., Hsueh C., Kuo T.T., Mounts A., Parashar U., Yang C.F., Guarner J., Ksiazek T.G., other authors. 2001; Pathologic studies of fatal cases in outbreak of hand, foot, and mouth disease. Emerg Infect Dis 7:146–148 [CrossRef]
     [Google Scholar]
  36. Sofroniew M.V., Vinters H.V. 2010; Astrocytes: biology and pathology. Acta Neuropathol 119:7–35 [View Article][PubMed]
     [Google Scholar]
  37. Song J.H., Bellail A., Tse M.C., Yong V.W., Hao C. 2006; Human astrocytes are resistant to Fas ligand and tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis. J Neurosci 26:3299–3308 [View Article][PubMed]
     [Google Scholar]
  38. Tung W.H., Sun C.C., Hsieh H.L., Wang S.W., Horng J.T., Yang C.M. 2007; EV71 induces VCAM-1 expression via PDGF receptor, PI3-K/Akt, p38 MAPK, JNK and NF-kappaB in vascular smooth muscle cells. Cell Signal 19:2127–2137 [View Article][PubMed]
     [Google Scholar]
  39. Tung W.-H., Hsieh H.-L., Yang C.-M. 2010; Enterovirus 71 induces COX-2 expression via MAPKs, NF-kappaB, and AP-1 in SK-N-SH cells: role of PGE2 in viral replication. Cell Signal 22:234–246 [View Article][PubMed]
     [Google Scholar]
  40. Wang S.M., Lei H.Y., Huang K.J., Wu J.M., Wang J.R., Yu C.K., Su I.J., Liu C.C. 2003; Pathogenesis of enterovirus 71 brainstem encephalitis in pediatric patients: roles of cytokines and cellular immune activation in patients with pulmonary edema. J Infect Dis 188:564–570 [View Article][PubMed]
     [Google Scholar]
  41. Wang S.-M., Lei H.-Y., Huang M.-C., Su L.-Y., Lin H.-C., Yu C.-K., Wang J.-L., Liu C.-C. 2006; Modulation of cytokine production by intravenous immunoglobulin in patients with enterovirus 71-associated brainstem encephalitis. J Clin Virol 37:47–52 [View Article][PubMed]
     [Google Scholar]
  42. Wang G., Zhang J., Li W., Xin G., Su Y., Gao Y., Zhang H., Lin G., Jiao X., Li K. 2008a; Apoptosis and proinflammatory cytokine responses of primary mouse microglia and astrocytes induced by human H1N1 and avian H5N1 influenza viruses. Cell Mol Immunol 5:113–120 [View Article][PubMed]
     [Google Scholar]
  43. Wang S.M., Lei H.Y., Yu C.K., Wang J.R., Su I.J., Liu C.C. 2008b; Acute chemokine response in the blood and cerebrospinal fluid of children with enterovirus 71-associated brainstem encephalitis. J Infect Dis 198:1002–1006 [View Article][PubMed]
     [Google Scholar]
  44. Wang S.M., Lei H.Y., Liu C.C. 2012; Cytokine immunopathogenesis of enterovirus 71 brain stem encephalitis. Clin Dev Immunol 2012: [View Article][PubMed]
     [Google Scholar]
  45. Wang C., Gao L., Jin Y., Cardona C.J., Xing Z. 2015; Regulation of host responses and viral replication by the mitogen-activated protein kinases in intestinal epithelial cells infected with Enterovirus 71. Virus Res 197:75–84 [View Article][PubMed]
     [Google Scholar]
  46. Yang Y., Wang H., Gong E., Du J., Zhao X., McNutt M.A., Wang S., Zhong Y., Gao Z., Zheng J. 2009; Neuropathology in 2 cases of fatal enterovirus type 71 infection from a recent epidemic in the People's Republic of China: a histopathologic, immunohistochemical, and reverse transcription polymerase chain reaction study. Hum Pathol 40:1288–1295 [View Article][PubMed]
     [Google Scholar]
  47. Yoneda S., Tanihara H., Kido N., Honda Y., Goto W., Hara H., Miyawaki N. 2001; Interleukin-1beta mediates ischemic injury in the rat retina. Exp Eye Res 73:661–667 [View Article][PubMed]
     [Google Scholar]
  48. Zhang Y., Zhu Z., Yang W., Ren J., Tan X., Wang Y., Mao N., Xu S., Zhu S., other authors. 2010; An emerging recombinant human enterovirus 71 responsible for the 2008 outbreak of hand foot and mouth disease in Fuyang city of China. Virol J 7:94 [View Article][PubMed]
     [Google Scholar]
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Intrinsic apoptosis and proinflammatory cytokines regulated in human astrocytes infected with enterovirus 71
J. Gen. Virol. 96, 3010 (2015); https://doi.org/10.1099/jgv.0.000235
/content/journal/jgv/10.1099/jgv.0.000235
/content/journal/jgv/10.1099/jgv.0.000235
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