Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

A FADD-dependent innate immune mechanism in mammalian cells

Nature volume 432, pages 401–405 (2004)Cite this article

Abstract

Vertebrate innate immunity provides a first line of defence against pathogens such as viruses and bacteria. Viral infection activates a potent innate immune response, which can be triggered by double-stranded (ds)RNA produced during viral replication1,2,3. Here, we report that mammalian cells lacking the death-domain-containing protein FADD4,5 are defective in intracellular dsRNA-activated gene expression, including production of type I (α/β) interferons, and are thus very susceptible to viral infection. The signalling pathway incorporating FADD is largely independent of Toll-like receptor 3 and the dsRNA-dependent kinase PKR, but seems to require receptor interacting protein 1 as well as Tank-binding kinase 1-mediated activation of the transcription factor IRF-3. The requirement for FADD in mammalian host defence is evocative of innate immune signalling in Drosophila, in which a FADD-dependent pathway responds to bacterial infection by activating the transcription of antimicrobial genes6. These data therefore suggest the existence of a conserved pathogen recognition pathway in mammalian cells that is essential for the optimal induction of type I interferons and other genes important for host defence.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: FADD-deficient MEFs are susceptible to VSV despite IFN pre-treatment.
Figure 2: Defective antiviral gene induction by intracellular dsRNA in the absence of FADD.
Figure 3: RIP1 deficiency mimics FADD ablation.
Figure 4: The antiviral pathway incorporating FADD signals via TBK-1 and IRF-3.

Similar content being viewed by others

References

  1. Takeda, K., Kaisho, T. & Akira, S. Toll-like receptors. Annu. Rev. Immunol. 21, 335–376 (2003)

    CAS  PubMed  Google Scholar 

  2. Medzhitov, R. & Janeway, C. Jr. The Toll receptor family and microbial recognition. Trends Microbiol. 8, 452–456 (2000)

    CAS  PubMed  Google Scholar 

  3. Taniguchi, T. & Takaoka, A. The interferon-α/β system in antiviral responses: a multimodal machinery of gene regulation by the IRF family of transcription factors. Curr. Opin. Immunol. 14, 111–116 (2002)

    CAS  PubMed  Google Scholar 

  4. Chinnaiyan, A. M., O'Rourke, K., Tewari, M. & Dixit, V. M. FADD, a novel death domain-containing protein, interacts with the death domain of Fas and initiates apoptosis. Cell 81, 505–512 (1995)

    CAS  PubMed  Google Scholar 

  5. Yeh, W. C. et al. FADD: essential for embryo development and signaling from some, but not all, inducers of apoptosis. Science 279, 1954–1958 (1998)

    ADS  CAS  PubMed  Google Scholar 

  6. Hoffmann, J. A. The immune response of Drosophila. Nature 426, 33–38 (2003)

    ADS  CAS  PubMed  Google Scholar 

  7. Stark, G. R., Kerr, I. M., Williams, B. R., Silverman, R. H. & Schreiber, R. D. How cells respond to interferons. Annu. Rev. Biochem. 67, 227–264 (1998)

    CAS  PubMed  Google Scholar 

  8. Balachandran, S. et al. Alpha/beta interferons potentiate virus-induced apoptosis through activation of the FADD/Caspase-8 death signaling pathway. J. Virol. 74, 1513–1523 (2000)

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Varfolomeev, E. E. et al. Targeted disruption of the mouse Caspase 8 gene ablates cell death induction by the TNF receptors, Fas/Apo1, and DR3 and is lethal prenatally. Immunity 9, 267–276 (1998)

    CAS  PubMed  Google Scholar 

  10. Muzio, M. et al. FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death-inducing signaling complex. Cell 85, 817–827 (1996)

    CAS  PubMed  Google Scholar 

  11. Levy, D. E. & Darnell, J. E. Jr. Stats: transcriptional control and biological impact. Nature Rev. Mol. Cell Biol. 3, 651–662 (2002)

    CAS  Google Scholar 

  12. Williams, B. R. PKR; a sentinel kinase for cellular stress. Oncogene 18, 6112–6120 (1999)

    CAS  PubMed  Google Scholar 

  13. Balachandran, S. et al. Essential role for the dsRNA-dependent protein kinase PKR in innate immunity to viral infection. Immunity 13, 129–141 (2000)

    CAS  PubMed  Google Scholar 

  14. Chu, W. M. et al. JNK2 and IKKβ are required for activating the innate response to viral infection. Immunity 11, 721–731 (1999)

    CAS  PubMed  Google Scholar 

  15. Diebold, S. S. et al. Viral infection switches non-plasmacytoid dendritic cells into high interferon producers. Nature 424, 324–328 (2003)

    ADS  CAS  PubMed  Google Scholar 

  16. Alexopoulou, L., Holt, A. C., Medzhitov, R. & Flavell, R. A. Recognition of double-stranded RNA and activation of NF-κB by Toll-like receptor 3. Nature 413, 732–738 (2001)

    ADS  CAS  PubMed  Google Scholar 

  17. Oshiumi, H., Matsumoto, M., Funami, K., Akazawa, T. & Seya, T. TICAM-1, an adaptor molecule that participates in Toll-like receptor 3-mediated interferon-β induction. Nature Immunol. 4, 161–167 (2003)

    CAS  Google Scholar 

  18. Yamamoto, M. et al. Role of adaptor TRIF in the MyD88-independent toll-like receptor signaling pathway. Science 301, 640–643 (2003)

    ADS  CAS  PubMed  Google Scholar 

  19. Hemmi, H. et al. The roles of two IκB kinase-related kinases in lipopolysaccharide and double stranded RNA signaling and viral infection. J. Exp. Med. 199, 1641–1650 (2004)

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Honda, K. et al. Selective contribution of IFN-α/β signaling to the maturation of dendritic cells induced by double-stranded RNA or viral infection. Proc. Natl Acad. Sci. USA 100, 10872–10877 (2003)

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  21. Hoebe, K. et al. Upregulation of costimulatory molecules induced by lipopolysaccharide and double-stranded RNA occurs by Trif-dependent and Trif-independent pathways. Nature Immunol. 4, 1223–1229 (2003)

    CAS  Google Scholar 

  22. Yoneyama, M. et al. The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses. Nature Immunol. 5, 730–737 (2004)

    CAS  Google Scholar 

  23. Leulier, F., Vidal, S., Saigo, K., Ueda, R. & Lemaitre, B. Inducible expression of double-stranded RNA reveals a role for dFADD in the regulation of the antibacterial response in Drosophila adults. Curr. Biol. 12, 996–1000 (2002)

    CAS  PubMed  Google Scholar 

  24. Naitza, S. et al. The Drosophila immune defense against gram-negative infection requires the death protein dFADD. Immunity 17, 575–581 (2002)

    CAS  PubMed  Google Scholar 

  25. Kelliher, M. A. et al. The death domain kinase RIP mediates the TNF-induced NF-κB signal. Immunity 8, 297–303 (1998)

    CAS  PubMed  Google Scholar 

  26. Meylan, E. et al. RIP1 is an essential mediator of Toll-like receptor 3-induced NF-κB activation. Nature Immunol. 5, 503–507 (2004)

    CAS  Google Scholar 

  27. Wathelet, M. G. et al. Virus infection induces the assembly of coordinately activated transcription factors on the IFN-β enhancer in vivo. Mol. Cell 1, 507–518 (1998)

    CAS  PubMed  Google Scholar 

  28. Sharma, S. et al. Triggering the interferon antiviral response through an IKK-related pathway. Science 300, 1148–1151 (2003)

    ADS  CAS  PubMed  Google Scholar 

  29. Fitzgerald, K. A. et al. IKKɛ and TBK1 are essential components of the IRF3 signaling pathway. Nature Immunol. 4, 491–496 (2003)

    CAS  Google Scholar 

  30. McWhirter, S. M. et al. IFN-regulatory factor 3-dependent gene expression is defective in Tbk1-deficient mouse embryonic fibroblasts. Proc. Natl Acad. Sci. USA 101, 233–238 (2004)

    ADS  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We are grateful to W.-C. Yeh, N. Kelliher, D. Wallach, J. Inoue, J. Durbin, J. Bell, K. Mossman, E. Harhaj, M. Karin, B. Williams and S. Akira for fibroblasts, and T. Maniatis, J. Hiscott and N. Reich and for plasmid constructs. We also thank G. Spruill, M. Fallahi and T. Andrew for technical assistance. This work was supported by DARPA.

Author information

Author notes

  1. Siddharth Balachandran and Emmanuel Thomas: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Microbiology and Immunology and Sylvester Comprehensive Cancer Center, University of Miami School of Medicine, Miami, Florida, 33136, USA

    Siddharth Balachandran, Emmanuel Thomas & Glen N. Barber

Authors
  1. Siddharth Balachandran
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Emmanuel Thomas
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Glen N. Barber
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Glen N. Barber.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Figure 1

Absence of FADD sensitizes cells to infection by Encephalomyocarditis Virus (EMCV) and Influenza Virus (FLU). (JPG 82 kb)

Supplementary Figure 2

Caspase 8 is not required for protection against VSV. (JPG 96 kb)

Supplementary Figure 3

Normal IFN signalling in the absence of FADD. (JPG 23 kb)

Supplementary Figure 4

Confirmation of a requirement for FADD for optimal induction of IFN-β following intracellular dsRNA treatment. (JPG 37 kb)

Supplementary Figure 5

TLR3 and PKR independent signalling by intracellular dsRNA. (JPG 72 kb)

Supplementary Figure 6

Analysis of RIPK1 -/- MEFs. (JPG 37 kb)

Supplementary Figure 7

Quantitation of cell death and virus yield from Tbk1/Ikkδ+/+ and Tbk1/Ikkδ-/-, as well as Irf3+/+ and Irf3-/- MEFs. (JPG 51 kb)

Supplementary Figure 8

FADD, RIP1 and IRF-3 are required for IFN-α production in 293 cells. (JPG 79 kb)

Supplementary Figure 9

The RIP/FADD/TBK-1 (RIFT) cascade. (JPG 29 kb)

Supplementary Figure Legends

Legends to accompany the above figures. (DOC 55 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Balachandran, S., Thomas, E. & Barber, G. A FADD-dependent innate immune mechanism in mammalian cells. Nature 432, 401–405 (2004). https://doi.org/10.1038/nature03124

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature03124

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing