1887

Journal of General Virology header logo

Volume 90, Issue 2

Influenza B mutant viruses with truncated NS1 proteins grow efficiently in Vero cells and are immunogenic in mice Free

Abstract

Contemporary influenza B virus strains were generated encoding C-terminally truncated NS1 proteins. Viable viruses containing the N-terminal 14, 38, 57 or 80 aa of the NS1 protein were rescued in Vero cells. The influenza B virus NS1-truncated mutants were impaired in their ability to counteract interferon (IFN) production, induce antiviral pro-inflammatory cytokines early after infection and show attenuated or restricted growth in IFN-competent hosts. In Vero cells, all of the mutant viruses replicated to high titres comparable to the wild-type influenza B virus. Mice that received a single, intranasal immunization of the NS1-truncated mutants elicited an antibody response and protection against wild-type virus challenge. Therefore, these NS1-truncated mutants should prove useful as potential candidates for live-attenuated influenza virus vaccines.

  • Received:
  • Accepted:
  • Published Online:
SGM
Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.006122-0
2009-02-01
2024-04-18
Loading full text...

Full text loading...

/deliver/fulltext/jgv/90/2/366.html?itemId=/content/journal/jgv/10.1099/vir.0.006122-0&mimeType=html&fmt=ahah

References

  1. 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]
  2. Dauber B., Heins G., Wolff T. 2004; The influenza B virus nonstructural NS1 protein is essential for efficient viral growth and antagonizes beta interferon induction. J Virol 78:1865–1872 [CrossRef]
     [Google Scholar]
  3. Dauber B., Schneider J., Wolff T. 2006; Double-stranded RNA binding of influenza B virus nonstructural NS1 protein inhibits protein kinase R but is not essential to antagonize production of alpha/beta interferon. J Virol 80:11667–11677 [CrossRef]
     [Google Scholar]
  4. Dittmann J., Stertz S., Grimm D., Steel J., García-Sastre A., Haller O., Kochs G. 2008; Influenza A virus strains differ in sensitivity to the antiviral action of the Mx-GTPase. J Virol 82:3624–3631 [CrossRef]
     [Google Scholar]
  5. Donelan N. R., Basler C. F., García-Sastre A. 2003; A recombinant influenza A virus expressing an RNA-binding-defective NS1 protein induces high levels of beta interferon and is attenuated in mice. J Virol 77:13257–13266 [CrossRef]
     [Google Scholar]
  6. Donelan N. R., Dauber B., Wang X., Basler C. F., Wolff T., García-Sastre A. 2004; The N- and C-terminal domains of the NS1 protein of influenza B virus can independently inhibit IRF-3 and beta interferon promoter activation. J Virol 78:11574–11582 [CrossRef]
     [Google Scholar]
  7. Du W., Maniatis T. 1992; An ATF/CREB binding site is required for virus induction of the human interferon beta gene [corrected]. Proc Natl Acad Sci U S A 89:2150–2154 [CrossRef]
     [Google Scholar]
  8. Egorov A., Brandt S., Sereinig S., Romanova J., Ferko B., Katinger D., Grassauer A., Alexandrova G., Katinger H., Muster T. 1998; Transfectant influenza A viruses with long deletions in the NS1 protein grow efficiently in Vero cells. J Virol 72:6437–6441
     [Google Scholar]
  9. Ferko B., Stasakova J., Romanova J., Kittel C., Sereinig S., Katinger H., Egorov A. 2004; Immunogenicity and protection efficacy of replication-deficient influenza A viruses with altered NS1 genes. J Virol 78:13037–13045 [CrossRef]
     [Google Scholar]
  10. Fitzgerald K. A., McWhirter S. M., Faia K. L., Rowe D. C., Latz E., Golenbock D. T., Coyle A. J., Liao S. M., Maniatis T. 2003; IKKϵ and TBK1 are essential components of the IRF3 signaling pathway. Nat Immunol 4:491–496
     [Google Scholar]
  11. García-Sastre A., Biron C. A. 2006; Type 1 interferons and the virus–host relationship: a lesson in détente. Science 312:879–882 [CrossRef]
     [Google Scholar]
  12. 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]
  13. 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]
  14. Holzinger D., Jorns C., Stertz S., Boisson-Dupuis S., Thimme R., Weidmann M., Casanova J. L., Haller O., Kochs G. 2007; Induction of MxA gene expression by influenza A virus requires type I or type III interferon signaling. J Virol 81:7776–7785 [CrossRef]
     [Google Scholar]
  15. Imai M., Kawasaki K., Odagiri T. 2008; Cytoplasmic domain of influenza B virus BM2 protein plays critical roles in production of infectious virus. J Virol 82:728–739 [CrossRef]
     [Google Scholar]
  16. Juang Y. T., Lowther W., Kellum M., Au W. C., Lin R., Hiscott J., Pitha P. M. 1998; Primary activation of interferon A and interferon B gene transcription by interferon regulatory factor 3. Proc Natl Acad Sci U S A 95:9837–9842 [CrossRef]
     [Google Scholar]
  17. Julkunen I., Sareneva T., Pirhonen J., Ronni T., Melen K., Matikainen S. 2001; Molecular pathogenesis of influenza A virus infection and virus-induced regulation of cytokine gene expression. Cytokine Growth Factor Rev 12:171–180 [CrossRef]
     [Google Scholar]
  18. Kaufmann A., Salentin R., Meyer R. G., Bussfeld D., Pauligk C., Fesq H., Hofmann P., Nain M., Gemsa D., Sprenger H. 2001; Defense against influenza A virus infection: essential role of the chemokine system. Immunobiology 204:603–613 [CrossRef]
     [Google Scholar]
  19. Kawai T., Takahashi K., Sato S., Coban C., Kumar H., Kato H., Ishii K. J., Takeuchi O., Akira S. 2005; IPS-1, an adaptor triggering RIG-I- and Mda5-mediated type I interferon induction. Nat Immunol 6:981–988 [CrossRef]
     [Google Scholar]
  20. Kim M. J., Latham A. G., Krug R. M. 2002; Human influenza viruses activate an interferon-independent transcription of cellular antiviral genes: outcome with influenza A virus is unique. Proc Natl Acad Sci U S A 99:10096–10101 [CrossRef]
     [Google Scholar]
  21. Kittel C., Ferko B., Kurz M., Voglauer R., Sereinig S., Romanova J., Stiegler G., Katinger H., Egorov A. 2005; Generation of an influenza A virus vector expressing biologically active human interleukin-2 from the NS gene segment. J Virol 79:10672–10677 [CrossRef]
     [Google Scholar]
  22. Lenschow D. J., Lai C., Frias-Staheli N., Giannakopoulos N. V., Lutz A., Wolff T., Osiak A., Levine B., Schmidt R. E. other authors 2007; IFN-stimulated gene 15 functions as a critical antiviral molecule against influenza, herpes, and Sindbis viruses. Proc Natl Acad Sci U S A 104:1371–1376 [CrossRef]
     [Google Scholar]
  23. Levy D. E., Marie I., Smith E., Prakash A. 2002; Enhancement and diversification of IFN induction by IRF-7-mediated positive feedback. J Interferon Cytokine Res 22:87–93 [CrossRef]
     [Google Scholar]
  24. Lin R., Heylbroeck C., Pitha P. M., Hiscott J. 1998; Virus-dependent phosphorylation of the IRF-3 transcription factor regulates nuclear translocation, transactivation potential, and proteasome-mediated degradation. Mol Cell Biol 18:2986–2996
     [Google Scholar]
  25. Lu Y., Qian X. Y., Krug R. M. 1994; The influenza virus NS1 protein: a novel inhibitor of pre-mRNA splicing. Genes Dev 8:1817–1828 [CrossRef]
     [Google Scholar]
  26. 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]
  27. Ludwig S., Wang X., Ehrhardt C., Zheng H., Donelan N., Planz O., Pleschka S., García-Sastre A., Heins G., Wolff T. 2002; The influenza A virus NS1 protein inhibits activation of Jun N-terminal kinase and AP-1 transcription factors. J Virol 76:11166–11171 [CrossRef]
     [Google Scholar]
  28. Ludwig S., Pleschka S., Planz O., Wolff T. 2006; Ringing the alarm bells: signalling and apoptosis in influenza virus infected cells. Cell Microbiol 8:375–386 [CrossRef]
     [Google Scholar]
  29. Marion R. M., Zurcher T., de la Luna S., Ortin J. 1997; Influenza virus NS1 protein interacts with viral transcription–replication complexes in vivo . J Gen Virol 78:2447–2451
     [Google Scholar]
  30. Matikainen S., Pirhonen J., Miettinen M., Lehtonen A., Govenius-Vintola C., Sareneva T., Julkunen I. 2000; Influenza A and Sendai viruses induce differential chemokine gene expression and transcription factor activation in human macrophages. Virology 276:138–147 [CrossRef]
     [Google Scholar]
  31. McWhirter S. M., Fitzgerald K. A., Rosains J., Rowe D. C., Golenbock D. T., Maniatis T. 2004; IFN-regulatory factor 3-dependent gene expression is defective in Tbk1-deficient mouse embryonic fibroblasts. Proc Natl Acad Sci U S A 101:233–238 [CrossRef]
     [Google Scholar]
  32. Meylan E., Curran J., Hofmann K., Moradpour D., Binder M., Bartenschlager R., Tschopp J. 2005; Cardif is an adaptor protein in the RIG-I antiviral pathway and is targeted by hepatitis C virus. Nature 437:1167–1172 [CrossRef]
     [Google Scholar]
  33. Min J. Y., Krug R. M. 2006; The primary function of RNA binding by the influenza A virus NS1 protein in infected cells: inhibiting the 2′-5′ oligo (A) synthetase/RNase L pathway. Proc Natl Acad Sci U S A 103:7100–7105 [CrossRef]
     [Google Scholar]
  34. Nemeroff M. E., Barabino S. M., Li Y., Keller W., Krug R. M. 1998; Influenza virus NS1 protein interacts with the cellular 30 kDa subunit of CPSF and inhibits 3′ end formation of cellular pre-mRNAs. Mol Cell 1:991–1000 [CrossRef]
     [Google Scholar]
  35. Palese P., Muster T., Zheng H., O'Neill R., García-Sastre A. 1999; Learning from our foes: a novel vaccine concept for influenza virus. Arch Virol Suppl 15:131–138
     [Google Scholar]
  36. Peters K. L., Smith H. L., Stark G. R., Sen G. C. 2002; IRF-3-dependent, NF κ B- and JNK-independent activation of the 561 and IFN- β genes in response to double-stranded RNA. Proc Natl Acad Sci U S A 99:6322–6327 [CrossRef]
     [Google Scholar]
  37. Pichlmair A., Schulz O., Tan C. P., Naslund T. I., Liljestrom P., Weber F., Reis. e Sousa C. 2006; RIG-I-mediated antiviral responses to single-stranded RNA bearing 5′-phosphates. Science 314:997–1001 [CrossRef]
     [Google Scholar]
  38. Proietti E., Bracci L., Puzelli S., Di Pucchio T., Sestili P., De Vincenzi E., Venditti M., Capone I., Seif I. other authors 2002; Type I IFN as a natural adjuvant for a protective immune response: lessons from the influenza vaccine model. J Immunol 169:375–383 [CrossRef]
     [Google Scholar]
  39. Qiu Y., Krug R. M. 1994; The influenza virus NS1 protein is a poly(A)-binding protein that inhibits nuclear export of mRNAs containing poly(A). J Virol 68:2425–2432
     [Google Scholar]
  40. Reed R., Cheng H. 2005; TREX, SR proteins and export of mRNA. Curr Opin Cell Biol 17:269–273 [CrossRef]
     [Google Scholar]
  41. Samuel C. E. 2001; Antiviral actions of interferons. Clin Microbiol Rev 14:778–809 [CrossRef]
     [Google Scholar]
  42. Sato M., Suemori H., Hata N., Asagiri M., Ogasawara K., Nakao K., Nakaya T., Katsuki M., Noguchi S. other authors 2000; Distinct and essential roles of transcription factors IRF-3 and IRF-7 in response to viruses for IFN - α / β gene induction. Immunity 13:539–548 [CrossRef]
     [Google Scholar]
  43. Seth R. B., Sun L., Ea C. K., Chen Z. J. 2005; Identification and characterization of MAVS, a mitochondrial antiviral signaling protein that activates NF- κ B and IRF 3. Cell 122:669–682 [CrossRef]
     [Google Scholar]
  44. Sharma S., tenOever B. R., Grandvaux N., Zhou G. P., Lin R., Hiscott J. 2003; Triggering the interferon antiviral response through an IKK-related pathway. Science 300:1148–1151 [CrossRef]
     [Google Scholar]
  45. Shaw M. W., Choppin P. W., Lamb R. A. 1983; A previously unrecognized influenza B virus glycoprotein from a bicistronic mRNA that also encodes the viral neuraminidase. Proc Natl Acad Sci U S A 80:4879–4883 [CrossRef]
     [Google Scholar]
  46. Shimizu K., Handa H., Nakada S., Nagata K. 1994; Regulation of influenza virus RNA polymerase activity by cellular and viral factors. Nucleic Acids Res 22:5047–5053 [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. Talon J., Horvath C. M., Polley R., Basler C. F., Muster T., Palese P., García-Sastre A. 2000a; Activation of interferon regulatory factor 3 is inhibited by the influenza A virus NS1 protein. J Virol 74:7989–7996 [CrossRef]
     [Google Scholar]
  49. Talon J., Salvatore M., O'Neill R. E., Nakaya Y., Zheng H., Muster T., García-Sastre A., Palese P. 2000b; Influenza A and B viruses expressing altered NS1 proteins: a vaccine approach. Proc Natl Acad Sci U S A 97:4309–4314 [CrossRef]
     [Google Scholar]
  50. Tovey M. G., Lallemand C., Thyphronitis G. 2008; Adjuvant activity of type I interferons. Biol Chem 389:541–545
     [Google Scholar]
  51. 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]
  52. 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 alpha/beta interferon. J Virol 74:11566–11573 [CrossRef]
     [Google Scholar]
  53. Wathelet M. G., Lin C. H., Parekh B. S., Ronco L. V., Howley P. M., Maniatis T. 1998; Virus infection induces the assembly of coordinately activated transcription factors on the IFN- β enhancer in vivo. Mol Cell 1:507–518 [CrossRef]
     [Google Scholar]
  54. Weaver B. K., Kumar K. P., Reich N. C. 1998; Interferon regulatory factor 3 and CREB-binding protein/p300 are subunits of double-stranded RNA-activated transcription factor DRAF1. Mol Cell Biol 18:1359–1368
     [Google Scholar]
  55. Wolff T., O'Neill R. E., Palese P. 1996; Interaction cloning of NS1-I, a human protein that binds to the nonstructural NS1 proteins of influenza A and B viruses. J Virol 70:5363–5372
     [Google Scholar]
  56. Wolff T., O'Neill R. E., Palese P. 1998; NS1-binding protein (NS1-BP): a novel human protein that interacts with the influenza A virus nonstructural NS1 protein is relocalized in the nuclei of infected cells. J Virol 72:7170–7180
     [Google Scholar]
  57. Wright P. F., Webster R. G. 2001; Orthomyxoviruses. In Fields Virology , 4th edn. pp 1533–1579Edited by Fields B. N., Knipe D. M. Philadelphia, PA: Lippincott Williams & Wilkins;
     [Google Scholar]
  58. Xu L. G., Wang Y. Y., Han K. J., Li L. Y., Zhai Z., Shu H. B. 2005; VISA is an adapter protein required for virus-triggered IFN- β signaling. Mol Cell 19:727–740 [CrossRef]
     [Google Scholar]
  59. Yoneyama M., Suhara W., Fukuhara Y., Fukuda M., Nishida E., Fujita T. 1998; Direct triggering of the type I interferon system by virus infection: activation of a transcription factor complex containing IRF-3 and CBP/p300. EMBO J 17:1087–1095 [CrossRef]
     [Google Scholar]
  60. Yoneyama M., Kikuchi M., Natsukawa T., Shinobu N., Imaizumi T., Miyagishi M., Taira K., Akira S., Fujita T. 2004; The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses. Nat Immunol 5:730–737 [CrossRef]
     [Google Scholar]
  61. Yuan W., Krug R. M. 2001; Influenza B virus NS1 protein inhibits conjugation of the interferon (IFN)-induced ubiquitin-like ISG15 protein. EMBO J 20:362–371 [CrossRef]
     [Google Scholar]
  62. Yuan W., Aramini J. M., Montelione G. T., Krug R. M. 2002; Structural basis for ubiquitin-like ISG 15 protein binding to the NS1 protein of influenza B virus: a protein–protein interaction function that is not shared by the corresponding N-terminal domain of the NS1 protein of influenza A virus. Virology 304:291–301 [CrossRef]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.006122-0
Loading
Influenza B mutant viruses with truncated NS1 proteins grow efficiently in Vero cells and are immunogenic in mice
J. Gen. Virol. 90, 366 (2009); https://doi.org/10.1099/vir.0.006122-0
/content/journal/jgv/10.1099/vir.0.006122-0
/content/journal/jgv/10.1099/vir.0.006122-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error