Oncolytic Viruses pp 217-238

Part of the Methods in Molecular Biology book series (MIMB, volume 797)

Evaluation of Innate Immune Signaling Pathways in Transformed Cells



Oncolytic viruses, the use of viruses to treat cancer, is emerging as a new option for cancer therapy. Oncolytic viruses, of both DNA and RNA origin, exhibit the ability to preferentially replicate in and kill cancer cells plausibly due to defects in innate immune signaling or translation regulation that are acquired during cellular transformation. Here, we review concepts and assays that describe how to analyze signaling pathways that govern the regulation of Type I IFN production as well as the induction of interferon-stimulated antiviral genes, events that are critical for mounting an effective antiviral response. The following procedures can be used to assess whether innate immune pathways that control antiviral host defense are defective in tumor cells – mechanisms that may help to explain viral oncolysis.

Key words

Innate immunity Interferon Oncolytic Host defense STING 


  1. 1.
    Barber GN. (2004) Vesicular stomatitis virus as an oncolytic vector Viral Immunol 17, 516–527.Google Scholar
  2. 2.
    Barber GN. (2005) VSV-tumor selective replication and protein translation Oncogene 24, 7710–7719.Google Scholar
  3. 3.
    Choi MK, Wang Z, Ban T, Yanai H, Lu Y, et al. (2009) A selective contribution of the RIG-I-like receptor pathway to type I interferon responses activated by cytosolic DNA Proc Natl Acad Sci USA 106, 17870–17875.Google Scholar
  4. 4.
    Yoneyama M, Fujita T. (2009) RNA recognition and signal transduction by RIG-I-like receptors Immunol Rev 227, 54–65.Google Scholar
  5. 5.
    Blasius AL, Beutler B. (2010) Intracellular toll-like receptors Immunity 32, 305–315.Google Scholar
  6. 6.
    Obuchi M, Fernandez M, Barber GN. (2003) Development of recombinant vesicular stomatitis viruses that exploit defects in host defense to augment specific oncolytic activity Journal of Virology 77, 8843–8856.Google Scholar
  7. 7.
    Akira S, Uematsu S, Takeuchi O. (2006) Pathogen recognition and innate immunity Cell 124, 783–801.Google Scholar
  8. 8.
    Wang L, Ligoxygakis P. (2006) Pathogen recognition and signalling in the Drosophila innate immune response Immunobiology 211, 251–261.Google Scholar
  9. 9.
    Silverman N, Maniatis T. (2001) NF-kappaB signaling pathways in mammalian and insect innate immunity Genes & Development 15, 2321–2342.Google Scholar
  10. 10.
    Leclerc V, Reichhart J-M. (2004) The immune response of Drosophila melanogaster Immunol Rev 198, 59–71.Google Scholar
  11. 11.
    Gottar M, Gobert V, Michel T, Belvin M, Duyk G, et al. (2002) The Drosophila immune response against Gram-negative bacteria is mediated by a peptidoglycan recognition protein Nature 416, 640–644.Google Scholar
  12. 12.
    Kawai T, Akira S. (2008) Toll-like receptor and RIG-I-like receptor signaling Ann N Y Acad Sci 1143, 1–20.Google Scholar
  13. 13.
    Alexopoulou L, Holt AC, Medzhitov R, Flavell RA. (2001) Recognition of double-stranded RNA and activation of NF-kappaB by Toll-like receptor 3 Nature 413, 732–738.Google Scholar
  14. 14.
    Taura M, Eguma A, Suico MA, Shuto T, Koga T, et al. (2008) p53 regulates Toll-like receptor 3 expression and function in human epithelial cell lines Molecular and Cellular Biology 28, 6557–6567.PubMedCrossRefGoogle Scholar
  15. 15.
    Lande R, Gilliet M. (2010) Plasmacytoid dendritic cells: key players in the initiation and regulation of immune responses Ann N Y Acad Sci 1183, 89–103.Google Scholar
  16. 16.
    Martinez J, Huang X, Yang Y. (2010) Toll-like receptor 8-mediated activation of murine plasmacytoid dendritic cells by vaccinia viral DNA Proceedings of the National Academy of Sciences 107, 6442–6447.CrossRefGoogle Scholar
  17. 17.
    Hemmi H, Takeuchi O, Kawai T, Kaisho T, Sato S, et al. (2000) A Toll-like receptor recognizes bacterial DNA Nature 408, 740–745.Google Scholar
  18. 18.
    Kawai T, Akira S. (2007) Antiviral signaling through pattern recognition receptors Journal of Biochemistry 141, 137–145.Google Scholar
  19. 19.
    Yamamoto M, Sato S, Hemmi H, Hoshino K, Kaisho T, et al. (2003) Role of adaptor TRIF in the MyD88-independent toll-like receptor ­signaling pathway Science 301, 640–643.Google Scholar
  20. 20.
    Oganesyan G, Saha SK, Guo B, He JQ, Shahangian A, et al. (2006) Critical role of TRAF3 in the Toll-like receptor-dependent and -independent antiviral response Nature 439, 208–211.PubMedCrossRefGoogle Scholar
  21. 21.
    Seya T, Shime H, Ebihara T, Oshiumi H, Matsumoto M. (2010) Pattern recognition receptors of innate immunity and their application to tumor immunotherapy. Cancer Sci 101(2), 313–320.Google Scholar
  22. 22.
    Kawai T, Akira S. (2007) TLR signaling Seminars in Immunology 19, 24–32.Google Scholar
  23. 23.
    Hornung V, Schlender J, Guenthner-Biller M, Rothenfusser S, Endres S, et al. (2004) Replication-dependent potent IFN-alpha induction in human plasmacytoid dendritic cells by a single-stranded RNA virus J Immunol 173, 5935–5943.Google Scholar
  24. 24.
    Uematsu S, Akira S. (2006) Toll-like receptors and innate immunity J Mol Med 84, 712–725.Google Scholar
  25. 25.
    Honda K, Yanai H, Negishi H, Asagiri M, Sato M, et al. (2005) IRF-7 is the master regulator of type-I interferon-dependent immune responses Nature 434, 772–777.PubMedCrossRefGoogle Scholar
  26. 26.
    Yoneyama M, Fujita T. (2007) RIG-I family RNA helicases: cytoplasmic sensor for antiviral innate immunity Cytokine & Growth Factor Reviews 18, 545–551.Google Scholar
  27. 27.
    Kato H, Takeuchi O, Sato S, Yoneyama M, Yamamoto M, et al. (2006) Differential roles of MDA5 and RIG-I helicases in the recognition of RNA viruses Nature 441, 101–105.PubMedCrossRefGoogle Scholar
  28. 28.
    Hornung V, Ellegast J, Kim S, Brzózka K, Jung A, et al. (2006) 5’-Triphosphate RNA is the ligand for RIG-I Science 314, 994–997.Google Scholar
  29. 29.
    Scott I, Norris KL. (2008) The mitochondrial antiviral signaling protein, MAVS, is cleaved during apoptosis Biochemical and Biophysical Research Communications 375, 101–106.Google Scholar
  30. 30.
    Meylan E, Curran J, Hofmann K, Moradpour D, Binder M, et al. (2005) Cardif is an adaptor protein in the RIG-I antiviral pathway and is targeted by hepatitis C virus Nature 437, 1167–1172.Google Scholar
  31. 31.
    Sharma S, tenOever BR, Grandvaux N, Zhou G-P, Lin R, et al. (2003) Triggering the interferon antiviral response through an IKK-related pathway Science 300, 1148–1151.Google Scholar
  32. 32.
    Balachandran S, Thomas E, Barber GN. (2004) A FADD-dependent innate immune mechanism in mammalian cells Nature 432, 401–405.Google Scholar
  33. 33.
    Balachandran S, Venkataraman T, Fisher PB, Barber GN. (2007) Fas-associated death domain-containing protein-mediated antiviral innate immune signaling involves the regulation of Irf7 J Immunol 178, 2429–2439.Google Scholar
  34. 34.
    Martin D, Gutkind JS. (2008) Human tumor-associated viruses and new insights into the molecular mechanisms of cancer Oncogene 27 Suppl 2, S31–42.Google Scholar
  35. 35.
    Stetson DB, Medzhitov R. (2006) Recognition of cytosolic DNA activates an IRF3-dependent innate immune response Immunity 24, 93–103.PubMedCrossRefGoogle Scholar
  36. 36.
    Wang Z, Choi MK, Ban T, Yanai H, Negishi H, et al. (2008) Regulation of innate immune responses by DAI (DLM-1/ZBP1) and other DNA-sensing molecules Proc Natl Acad Sci USA 105, 5477–5482.Google Scholar
  37. 37.
    Ishikawa H, Barber GN. (2008) STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling Nature 455, 674–678.Google Scholar
  38. 38.
    Ishikawa H, Ma Z, Barber GN. (2009) STING regulates intracellular DNA-mediated, type I interferon-dependent innate immunity Nature 461, 788–792.PubMedCrossRefGoogle Scholar
  39. 39.
    Constantinescu SN, Girardot M, Pecquet C. (2008) Mining for JAK-STAT mutations in cancer Trends Biochem Sci 33, 122–131.Google Scholar
  40. 40.
    Zou W, Kim J-H, Handidu A, Li X, Kim KI, et al. (2007) Microarray analysis reveals that Type I interferon strongly increases the expression of immune-response related genes in Ubp43 (Usp18) deficient macrophages Biochemical and Biophysical Research Communications 356, 193–199.PubMedCrossRefGoogle Scholar
  41. 41.
    Platanias LC, Fish EN. (1999) Signaling pathways activated by interferons Exp Hematol 27, 1583–1592.Google Scholar
  42. 42.
    Darnell JE. (1997) STATs and gene regulation Science 277, 1630–1635.Google Scholar
  43. 43.
    Minegishi Y, Saito M, Morio T, Watanabe K, Agematsu K, et al. (2006) Human tyrosine kinase 2 deficiency reveals its requisite roles in multiple cytokine signals involved in innate and acquired immunity Immunity 25: 745–755.PubMedCrossRefGoogle Scholar
  44. 44.
    Schindler C, Levy DE, Decker T. (2007) JAK-STAT signaling: from interferons to cytokinesJ Biol Chem 282, 20059–20063.Google Scholar
  45. 45.
    Platanias LC. (2005) Mechanisms of type-I- and type-II-interferon-mediated signalling Nat Rev Immunol 5, 375–386.PubMedCrossRefGoogle Scholar
  46. 46.
    Joshi S, Kaur S, Kroczynska B, Platanias LC. (2010) Mechanisms of mRNA translation of interferon stimulated genes. Cytokine.Google Scholar
  47. 47.
    Balachandran S, Barber GN. (2004) Defective translational control facilitates vesicular stomatitis virus oncolysis Cancer Cell 5, 51–65.Google Scholar
  48. 48.
    Pfeifer I, Elsby R, Fernandez M, Faria PA, Nussenzveig DR, et al. (2008) NFAR-1 and -2 modulate translation and are required for efficient host defense Proc Natl Acad Sci USA 105, 4173–4178.Google Scholar
  49. 49.
    Colina R, Costa-Mattioli M, Dowling RJO, Jaramillo M, Tai L-H, et al. (2008) Translational control of the innate immune response through IRF-7 Nature 452, 323–328.Google Scholar
  50. 50.
    Takaoka A, Hayakawa S, Yanai H, Stoiber D, Negishi H, et al. (2003) Integration of interferon-alpha/beta signalling to p53 responses in tumour suppression and antiviral defence Nature 424, 516–523.PubMedCrossRefGoogle Scholar
  51. 51.
    Pedersen IM, Cheng G, Wieland S, Volinia S, Croce CM, et al. (2007) Interferon modulation of cellular microRNAs as an antiviral mechanism Nature 449, 919–922.Google Scholar
  52. 52.
    Ohno M, Natsume A, Kondo Y, Iwamizu H, Motomura K, et al. (2009) The modulation of microRNAs by type I IFN through the activation of signal transducers and activators of transcription 3 in human glioma Mol Cancer Res 7, 2022–2030.PubMedCrossRefGoogle Scholar
  53. 53.
    Jarmalaite S, Andrekute R, Scesnaite A, Suziedelis K, Husgafvel-Pursiainen K, et al. (2010) Promoter hypermethylation in tumour suppressor genes and response to interleukin-2 treatment in bladder cancer: a pilot study J Cancer Res Clin Oncol 136, 847–854.PubMedCrossRefGoogle Scholar
  54. 54.
    Marozin S, Altomonte J, Stadler F, Thasler WE, Schmid RM, et al. (2008) Inhibition of the IFN-beta response in hepatocellular carcinoma by alternative spliced isoform of IFN regulatory factor-3 Mol Ther 16, 1789–1797.Google Scholar
  55. 55.
    Jee CD, Kim MA, Jung EJ, Kim J, Kim WH. (2009) Identification of genes epigenetically silenced by CpG methylation in human gastric carcinoma Eur J Cancer 45, 1282–1293.Google Scholar
  56. 56.
    Almeida S, Maillard C, Itin P, Hohl D, Huber M. (2008) Five new CYLD mutations in skin appendage tumors and evidence that aspartic acid 681 in CYLD is essential for deubiquitinase activity J Invest Dermatol 128, 587–593.PubMedGoogle Scholar
  57. 57.
    Critchley-Thorne RJ, Yan N, Nacu S, Weber J, Holmes SP, et al. (2007) Down-regulation of the interferon signaling pathway in T lymphocytes from patients with metastatic melanoma PLoS Med 4, e176.Google Scholar
  58. 58.
    Pansky A, Hildebrand P, Fasler-Kan E, Baselgia L, Ketterer S, et al. (2000) Defective Jak-STAT signal transduction pathway in melanoma cells resistant to growth inhibition by interferon-alpha Int J Cancer 85, 720–725.Google Scholar
  59. 59.
    Mullighan CG, Zhang J, Harvey RC, Collins-Underwood JR, Schulman BA, et al. (2009) JAK mutations in high-risk childhood acute lymphoblastic leukemia Proc Natl Acad Sci USA 106, 9414–9418.Google Scholar
  60. 60.
    van Eyndhoven WG, Gamper CJ, Cho E, Mackus WJ, Lederman S. (1999) TRAF-3 mRNA splice-deletion variants encode isoforms that induce NF-kappaB activation Molecular Immunology 36, 647–658.PubMedCrossRefGoogle Scholar
  61. 61.
    Nagel I, Bug S, Tönnies H, Ammerpohl O, Richter J, et al. (2009) Biallelic inactivation of TRAF3 in a subset of B-cell lymphomas with interstitial del(14)(q24.1q32.33) Leukemia 23, 2153–2155.PubMedCrossRefGoogle Scholar
  62. 62.
    Barbie DA, Tamayo P, Boehm JS, Kim SY, Moody SE, et al. (2009) Systematic RNA interference reveals that oncogenic KRAS-driven cancers require TBK1 Nature 462, 108–112.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.University of Miami School of MedicineMiamiUSA

Personalised recommendations