1887

Abstract

The NS1 protein is known to suppress immune responses in influenza virus-infected hosts. However, the role of NS1 in apoptosis in infected cells is disputed. In this study, through the use of a mutant A/pheasant/California/2373/1998 (H9N2) avian influenza virus (AIV) with a truncated NS1, we have demonstrated that a functional NS1 protein suppresses the induction of interferons in chicken macrophages. However, NS1 appeared to be irrelevant to the regulation of cytokines interleukin (IL)-1 and IL-6, indicating that distinct mechanisms may be employed in the regulation of antiviral and proinflammatory cytokines in chicken immune cells. Our study also showed that this H9N2 AIV induced apoptosis extrinsically through the Fas/Fas ligand (FasL)-mediated pathway. We found that NS1 suppressed the apoptotic process through suppression of the induction of FasL, but not tumour necrosis factor- or TNF-related apoptosis-inducing ligand. Furthermore, our data indicated that the disruption of a potential binding site for the p85 subunit of phosphoinositide 3-kinase in the carboxyl terminus of NS1, while having no effect on the regulation of IFN induction, may contribute to the suppression of Fas/FasL-mediated apoptosis. Therefore, suppression of Fas/FasL-mediated apoptosis by NS1 is one of the critical mechanisms necessary to increase infectivity in AIV-infected chicken macrophages.

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2009-05-01
2024-03-28
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References

  1. Bergmann M., García-Sastre A., Carnero E., Pehamberger H., Wolff K., Palese P., Muster T. 2000; Influenza virus NS1 protein counteracts PKR-mediated inhibition of replication. J Virol 74:6203–6206 [CrossRef]
    [Google Scholar]
  2. Bergmann M., Romirer I., Sachet M., Fleischhacker R., García-Sastre A., Palese P., Wolff K., Pehamberger H., Jakesz R., Muster T. 2001; A genetically engineered influenza A virus with ras -dependent oncolytic properties. Cancer Res 61:8188–8193
    [Google Scholar]
  3. Brunet A., Bonni A., Zigmond M. J., Lin M. Z., Juo P., Hu L. S., Anderson M. J., Arden K. C., Blenis J., Greenberg M. E. 1999; Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell 96:857–868 [CrossRef]
    [Google Scholar]
  4. Brydon E. W. A., Smith H., Sweet C. 2003; Influenza A virus-induced apoptosis in bronchiolar epithelial (NCI-H292) cells limits pro-inflammatory cytokine release. J Gen Virol 84:2389–2400 [CrossRef]
    [Google Scholar]
  5. Brydon E. W. A., Morris S. J., Sweet C. 2005; Role of apoptosis and cytokines in influenza virus morbidity. FEMS Microbiol Rev 29:837–850 [CrossRef]
    [Google Scholar]
  6. Cardone M. H., Roy N., Stennicke H. R., Salvesen G. S., Franke T. F., Stanbridge E., Frisch S., Reed J. C. 1998; Regulation of cell death protease caspase-9 by phosphorylation. Science 282:1318–1321 [CrossRef]
    [Google Scholar]
  7. Chanturiya A. N., Basanez G., Schubert U., Henklein P., Yewdell J. W., Zimmerberg J. 2004; PB1-F2, an influenza A virus-encoded proapoptotic mitochondrial protein, creates variably sized pores in planar lipid membranes. J Virol 78:6304–6312 [CrossRef]
    [Google Scholar]
  8. Chen W., Calvo P. A., Malide D., Gibbs J., Schubert U., Bacik I., Basta S., O'Neill R., Schickli J. other authors 2001; A novel influenza A virus mitochondrial protein that induces cell death. Nat Med 7:1306–1312 [CrossRef]
    [Google Scholar]
  9. Ciechomska I., Pyrzynska B., Kazmierczak P., Kaminska B. 2003; Inhibition of Akt kinase signalling and activation of Forkhead are indispensable for upregulation of FasL expression in apoptosis of glioma cells. Oncogene 22:7617–7627 [CrossRef]
    [Google Scholar]
  10. Datta S. R., Dudek H., Tao X., Masters S., Fu H., Gotoh Y., Greenberg M. E. 1997; Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell 91:231–241 [CrossRef]
    [Google Scholar]
  11. del Peso L., González-García M., Page C., Herrera R., Nuñez G. 1997; Interleukin-3-induced phosphorylation of BAD through the protein kinase Akt. Science 278:687–689 [CrossRef]
    [Google Scholar]
  12. Ehrhardt C., Wolff T., Ludwig S. 2007a; Activation of phosphatidylinositol 3-kinase signaling by the nonstructural NS1 protein is not conserved among type A and B influenza viruses. J Virol 81:12097–12100 [CrossRef]
    [Google Scholar]
  13. Ehrhardt C., Wolff T., Pleschka S., Planz O., Beermann W., Bode J. G., Schmolke M., Ludwig S. 2007b; Influenza A virus NS1 protein activates the PI3K/Akt pathway to mediate antiapoptotic signaling responses. J Virol 81:3058–3067 [CrossRef]
    [Google Scholar]
  14. Fesq H., Bacher M., Nain M., Gemsa D. 1994; Programmed cell death (apoptosis) in human monocytes infected by influenza A virus. Immunobiology 190:175–182 [CrossRef]
    [Google Scholar]
  15. Flory E., Kunz M., Scheller C., Jassoy C., Stauber R., Rapp U. R., Ludwig S. 2000; Influenza virus-induced NF- κ B-dependent gene expression is mediated by overexpression of viral proteins and involves oxidative radicals and activation of I κ B kinase. J Biol Chem 275:8307–8314 [CrossRef]
    [Google Scholar]
  16. García-Sastre A., Egorov A., Matassov D., Brandt S., Levy D. E., Durbin J. E., Palese P., Muster T. 1998; Influenza A virus lacking the NS1 gene replicates in interferon-deficient systems. Virology 252:324–330 [CrossRef]
    [Google Scholar]
  17. Gaush C. R., Smith T. F. 1968; Replication and plaque assay of influenza virus in an established line of canine kidney cells. Appl Microbiol 16:588–594
    [Google Scholar]
  18. Geiss G. K., Salvatore M., Tumpey T. M., Carter V. S., Wang X., Basler C. F., Taubenberger J. K., Bumgarner R. E., Palese P. other authors 2002; Cellular transcriptional profiling in influenza A virus-infected lung epithelial cells: the role of the nonstructural NS1 protein in the evasion of the host innate defense and its potential contribution to pandemic influenza. Proc Natl Acad Sci U S A 99:10736–10741 [CrossRef]
    [Google Scholar]
  19. Hale B. G., Jackson D., Chen Y. H., Lamb R. A., Randall R. E. 2006; Influenza A virus NS1 protein binds p85 β and activates phosphatidylinositol-3-kinase signaling. Proc Natl Acad Sci U S A 103:14194–14199 [CrossRef]
    [Google Scholar]
  20. Hale B. G., Batty I. H., Downes C. P., Randall R. E. 2008; Binding of influenza A virus NS1 protein to the inter-SH2 domain of p85 suggests a novel mechanism for phosphoinositide 3-kinase activation. J Biol Chem 283:1372–1380 [CrossRef]
    [Google Scholar]
  21. Hatada E., Saito S., Fukuda R. 1999; Mutant influenza viruses with a defective NS1 protein cannot block the activation of PKR in infected cells. J Virol 73:2425–2433
    [Google Scholar]
  22. Heikkinen L. S., Kazlauskas A., Melén K., Wagner R., Ziegler T., Julkunen I., Saksela K. 2008; Avian and 1918 Spanish influenza A virus NS1 proteins bind to Crk/CrkL Src homology 3 domains to activate host cell signaling. J Biol Chem 283:5719–5727
    [Google Scholar]
  23. Hinshaw V. S., Olsen C. W., Dybdahl-Sissoko N., Evans D. 1994; Apoptosis: a mechanism of cell killing by influenza A and B viruses. J Virol 68:3667–3673
    [Google Scholar]
  24. Hirst G. K. 1948; The nature of hemagglutination by viruses. Harvey Lect 44:84–98
    [Google Scholar]
  25. Hoffmann E., Stech J., Guan Y., Webster R. G., Perez D. R. 2001; Universal primer set for the full-length amplification of all influenza A viruses. Arch Virol 146:2275–2289 [CrossRef]
    [Google Scholar]
  26. Hoffmann E., Krauss S., Perez D., Webby R., Webster R. G. 2002; Eight-plasmid system for rapid generation of influenza virus vaccines. Vaccine 20:3165–3170 [CrossRef]
    [Google Scholar]
  27. Hofmann P., Sprenger H., Kaufmann A., Bender A., Hasse C., Nain M., Gemsa D. 1997; Susceptibility of mononuclear phagocytes to influenza A virus infection and possible role in the antiviral response. J Leukoc Biol 61:408–414
    [Google Scholar]
  28. Ito T., Kobayashi Y., Morita T., Horimoto T., Kawaoka Y. 2002; Virulent influenza A viruses induce apoptosis in chickens. Virus Res 84:27–35 [CrossRef]
    [Google Scholar]
  29. Lam W. Y., Tang J. W., Yeung A. C., Chiu L. C., Sung J. J., Chan P. K. 2008; Avian influenza virus A/HK/483/97(H5N1) NS1 protein induces apoptosis in human airway epithelial cells. J Virol 82:2741–2751 [CrossRef]
    [Google Scholar]
  30. Li S., Min J. Y., Krug R. M., Sen G. C. 2006; Binding of the influenza A virus NS1 protein to PKR mediates the inhibition of its activation by either PACT or double-stranded RNA. Virology 349:13–21 [CrossRef]
    [Google Scholar]
  31. Lowy R. J. 2003; Influenza virus induction of apoptosis by intrinsic and extrinsic mechanisms. Int Rev Immunol 22:425–449 [CrossRef]
    [Google Scholar]
  32. Lu Y., Wambach M., Katze M. G., Krug R. M. 1995; Binding of the influenza virus NS1 protein to double-stranded RNA inhibits the activation of the protein kinase that phosphorylates the elF-2 translation initiation factor. Virology 214:222–228 [CrossRef]
    [Google Scholar]
  33. Min J. Y., Li S., Sen G. C., Krug R. M. 2007; A site on the influenza A virus NS1 protein mediates both inhibition of PKR activation and temporal regulation of viral RNA synthesis. Virology 363:236–243 [CrossRef]
    [Google Scholar]
  34. Mok C. K., Lee D. C., Cheung C. Y., Peiris M., Lau A. S. 2007; Differential onset of apoptosis in influenza A virus H5N1- and H1N1-infected human blood macrophages. J Gen Virol 88:1275–1280 [CrossRef]
    [Google Scholar]
  35. Mori I., Komatsu T., Takeuchi K., Nakakuki K., Sudo M., Kimura Y. 1995; In vivo induction of apoptosis by influenza virus. J Gen Virol 76:2869–2873 [CrossRef]
    [Google Scholar]
  36. Morris S. J., Price G. E., Barnett J. M., Hiscox S. A., Smith H., Sweet C. 1999; Role of neuraminidase in influenza virus-induced apoptosis. J Gen Virol 80:137–146
    [Google Scholar]
  37. Morris S. J., Smith H., Sweet C. 2002; Exploitation of the Herpes simplex virus translocating protein VP22 to carry influenza virus proteins into cells for studies of apoptosis: direct confirmation that neuraminidase induces apoptosis and indications that other proteins may have a role. Arch Virol 147:961–979 [CrossRef]
    [Google Scholar]
  38. Morris S. J., Nightingale K., Smith H., Sweet C. 2005; Influenza A virus-induced apoptosis is a multifactorial process: exploiting reverse genetics to elucidate the role of influenza A virus proteins in virus-induced apoptosis. Virology 335:198–211 [CrossRef]
    [Google Scholar]
  39. Pahl H. L., Baeuerle P. A. 1995; Expression of influenza virus hemagglutinin activates transcription factor NF- κ B. J Virol 69:1480–1484
    [Google Scholar]
  40. Perdue M. L., Swayne D. E. 2005; Public health risk from avian influenza viruses. Avian Dis 49:317–327 [CrossRef]
    [Google Scholar]
  41. Perez D. R., Sorrell E. M., Donis R. O. 2005; Avian influenza: an omnipresent pandemic threat. Pediatr Infect Dis J 24:S208–S216 (discussion S215 [CrossRef]
    [Google Scholar]
  42. Rath N. C., Parcells M. S., Xie H., Santin E. 2003; Characterization of a spontaneously transformed chicken mononuclear cell line. Vet Immunol Immunopathol 96:93–104 [CrossRef]
    [Google Scholar]
  43. Ronni T., Matikainen S., Sareneva T., Melén K., Pirhonen J., Keskinen P., Julkunen I. 1997; Regulation of IFN- α / β , MxA, 2′,5′-oligoadenylate synthetase, and HLA gene expression in influenza A-infected human lung epithelial cells. J Immunol 158:2363–2374
    [Google Scholar]
  44. Schultz-Cherry S., Dybdahl-Sissoko N., Neumann G., Kawaoka Y., Hinshaw V. S. 2001; Influenza virus NS1 protein induces apoptosis in cultured cells. J Virol 75:7875–7881 [CrossRef]
    [Google Scholar]
  45. Shin Y. K., Li Y., Liu Q., Anderson D. H., Babiuk L. A., Zhou Y. 2007; SH3 binding motif 1 in influenza A virus NS1 protein is essential for PI3K/Akt signaling pathway activation. J Virol 81:12730–12739 [CrossRef]
    [Google Scholar]
  46. Song H., Nieto G. R., Perez D. R. 2007; A new generation of modified live-attenuated avian influenza viruses using a two-strategy combination as potential vaccine candidates. J Virol 81:9238–9248 [CrossRef]
    [Google Scholar]
  47. Stasakova J., Ferko B., Kittel C., Sereinig S., Romanova J., Katinger H., Egorov A. 2005; Influenza A mutant viruses with altered NS1 protein function provoke caspase-1 activation in primary human macrophages, resulting in fast apoptosis and release of high levels of interleukins 1 β and 18. J Gen Virol 86:185–195 [CrossRef]
    [Google Scholar]
  48. Sternsdorf T., Jensen K., Zuchner D., Will H. 1997; Cellular localization, expression, and structure of the nuclear dot protein 52. J Cell Biol 138:435–448 [CrossRef]
    [Google Scholar]
  49. Takizawa T. 1996; Mechanism of the induction of apoptosis by influenza virus infection. Nippon Rinsho 54:1836–1841
    [Google Scholar]
  50. Takizawa T., Matsukawa S., Higuchi Y., Nakamura S., Nakanishi Y., Fukuda R. 1993; Induction of programmed cell death (apoptosis) by influenza virus infection in tissue culture cells. J Gen Virol 74:2347–2355 [CrossRef]
    [Google Scholar]
  51. Takizawa T., Ohashi K., Nakanishi Y. 1996; Possible involvement of double-stranded RNA-activated protein kinase in cell death by influenza virus infection. J Virol 70:8128–8132
    [Google Scholar]
  52. Talon J., Horvath C. M., Polley R., Basler C. F., Muster T., Palese P., García-Sastre A. 2000; Activation of interferon regulatory factor 3 is inhibited by the influenza A virus NS1 protein. J Virol 74:7989–7996 [CrossRef]
    [Google Scholar]
  53. Uiprasertkul M., Kitphati R., Puthavathana P., Kriwong R., Kongchanagul A., Ungchusak K., Angkasekwinai S., Chokephaibulkit K., Srisook K. other authors 2007; Apoptosis and pathogenesis of avian influenza A (H5N1) virus in humans. Emerg Infect Dis 13:708–712 [CrossRef]
    [Google Scholar]
  54. Wan H., Perez D. R. 2007; Amino acid 226 in the hemagglutinin of H9N2 influenza viruses determines cell tropism and replication in human airway epithelial cells. J Virol 81:5181–5191 [CrossRef]
    [Google Scholar]
  55. Wang W., Krug R. M. 1996; The RNA-binding and effector domains of the viral NS1 protein are conserved to different extents among influenza A and B viruses. Virology 223:41–50 [CrossRef]
    [Google Scholar]
  56. Wang X., Li M., Zheng H., Muster T., Palese P., Beg A. A., García-Sastre A. 2000; Influenza A virus NS1 protein prevents activation of NF- κ B and induction of α / β interferon. J Virol 74:11566–11573 [CrossRef]
    [Google Scholar]
  57. Woolcock P. R., Suarez D. L., Kuney D. 2003; Low-pathogenicity avian influenza virus (H6N2) in chickens in California, 2000–02. Avian Dis 47:872–881 [CrossRef]
    [Google Scholar]
  58. Wurzer W. J., Ehrhardt C., Pleschka S., Berberich-Siebelt F., Wolff T., Walczak H., Planz O., Ludwig S. 2004; NF- κ B-dependent induction of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and Fas/FasL is crucial for efficient influenza virus propagation. J Biol Chem 279:30931–30937 [CrossRef]
    [Google Scholar]
  59. Xing Z., Conway E. M., Kang C., Winoto A. 2004; Essential role of survivin, an inhibitor of apoptosis protein, in T cell development, maturation, and homeostasis. J Exp Med 199:69–80
    [Google Scholar]
  60. Xing Z., Cardona C. J., Li J., Dao N., Tran T., Andrada J. 2008; Modulation of the immune responses in chickens by low-pathogenicity avian influenza virus H9N2. J Gen Virol 89:1288–1299 [CrossRef]
    [Google Scholar]
  61. Zhirnov O. P., Konakova T. E., Wolff T., Klenk H. D. 2002; NS1 protein of influenza A virus down-regulates apoptosis. J Virol 76:1617–1625 [CrossRef]
    [Google Scholar]
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