Skip to main content

Thank you for visiting 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.

Fas triggers an alternative, caspase-8–independent cell death pathway using the kinase RIP as effector molecule


Cell death is achieved by two fundamentally different mechanisms: apoptosis and necrosis. Apoptosis is dependent on caspase activation, whereas the caspase-independent necrotic signaling pathway remains largely uncharacterized. We show here that Fas kills activated primary T cells efficiently in the absence of active caspases, which results in necrotic morphological changes and late mitochondrial damage but no cytochrome c release. This Fas ligand–induced caspase-independent death is absent in T cells that are deficient in either Fas-associated death domain (FADD) or receptor-interacting protein (RIP). RIP is also required for necrotic death induced by tumor necrosis factor (TNF) and TNF-related apoptosis-inducing ligand (TRAIL). In contrast to its role in nuclear factor κB activation, RIP requires its own kinase activity for death signaling. Thus, Fas, TRAIL and TNF receptors can initiate cell death by two alternative pathways, one relying on caspase-8 and the other dependent on the kinase RIP.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: FasL-induced and activation-induced cell death in primary human and murine T cells is insensitive to the caspase inhibitor zVAD-fmk.
Figure 2: FasL-mediated, caspase-independent cell death in T cells requires FADD.
Figure 3: RIP binds to Fas and signals necrotic T cell death.
Figure 4: FasL-induced, caspase-independent T cell death is associated with necrotic morphological changes that are independent of cytochrome c release.
Figure 5: TNF and TRAIL signal caspase-independent cell death in T cells.
Figure 6: Putative model of death receptor-induced necrosis.


  1. 1

    Kerr, J., Wyllie, A. H. & Currie, A. R. Apoptosis a basic biological phenomenon with wide ranging implications in tissue kinetics. Br. J. Cancer 68, 239–257 (1972).

    Article  Google Scholar 

  2. 2

    Fiers, W., Beyaert, R., Declercq, W. & Vandenabeele, P. More than one way to die: apoptosis, necrosis and reactive oxygen damage. Oncogene 18, 7719–7730 (1999).

    CAS  Article  Google Scholar 

  3. 3

    Scaffidi, C., Kirchhoff, S., Krammer, P. H. & Peter, M. E. Apoptosis signaling in lymphocytes. Curr. Opin. Immunol. 11, 277–285 (1999).

    CAS  Article  Google Scholar 

  4. 4

    Vercammen, D. et al. Dual signaling of the Fas receptor: initiation of both apoptotic and necrotic cell death pathways. J. Exp. Med. 188, 919–930 (1998).

    CAS  Article  Google Scholar 

  5. 5

    Kawahara, A., Ohsawa, Y., Matsumura, H., Uchiyama, Y. & Nagata, S. Caspase-independent cell killing by Fas-associated protein with death domain. J. Cell Biol. 143, 1353–1360 (1998).

    CAS  Article  Google Scholar 

  6. 6

    Schneider, P. et al. Conversion of membrane-bound Fas(CD95) ligand to its soluble form is associated with downregulation of its proapoptotic activity and loss of liver toxicity. J. Exp. Med. 187, 1205–1213 (1998).

    CAS  Article  Google Scholar 

  7. 7

    Klas, C., Debatin, K. M., Jonker, R. R. & Krammer, P. H. Activation interferes with the APO-1 pathway in mature human T cells. Int. Immunol. 5, 625–630 (1993).

    CAS  Article  Google Scholar 

  8. 8

    Irmler, M. et al. Inhibition of death receptor signals by cellular FLIP. Nature 388, 190–195 (1997).

    CAS  Article  Google Scholar 

  9. 9

    Smith, K. G., Strasser, A. & Vaux, D. L. CrmA expression in T lymphocytes of transgenic mice inhibits CD95 (Fas/APO-1)-transduced apoptosis, but does not cause lymphadenopathy or autoimmune disease. EMBO J. 15, 5167–5176 (1996).

    CAS  Article  Google Scholar 

  10. 10

    Juo, P., Kuo, C. J., Yuan, J. & Blenis, J. Essential requirement for caspase-8/FLICE in the initiation of the Fas- induced apoptotic cascade. Curr. Biol. 8, 1001–1008 (1998).

    CAS  Article  Google Scholar 

  11. 11

    Juo, P. et al. FADD is required for multiple signaling events downstream of the receptor Fas. Cell Growth Differ. 10, 797–804 (1999).

    CAS  PubMed  Google Scholar 

  12. 12

    Kataoka, T. et al. The caspase-8 inhibitor FLIP promotes activation of NF-κB and erk signaling pathways. Curr. Biol. 10, 640–648 (2000).

    CAS  Article  Google Scholar 

  13. 13

    Stanger, B. Z., Leder, P., Lee, T. H., Kim, E. & Seed, B. RIP - a novel protein containing a death domain that interacts with Fas/Apo-1 (CD95) in yeast and causes cell death. Cell 81, 513–523 (1995).

    CAS  Article  Google Scholar 

  14. 14

    Ting, A. T., Pimentel-Muinos, F. X. & Seed, B. RIP mediates tumor necrosis factor receptor 1 activation of NF-κB but not Fas/APO-1-initiated apoptosis. EMBO J. 15, 6189–6196 (1996).

    CAS  Article  Google Scholar 

  15. 15

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

    CAS  Article  Google Scholar 

  16. 16

    Grimm, S., Stanger, B. Z. & Leder, P. RIP and FADD: two “death domain”-containing proteins can induce apoptosis by convergent, but dissociable, pathways. Proc. Natl Acad. Sci. USA 93, 10923–10927 (1996).

    CAS  Article  Google Scholar 

  17. 17

    Hsu, H., Huang, J., Shu, H. B., Baichwal, V. & Goeddel, D. V. TNF-dependent recruitment of the protein kinase RIP to the TNF receptor-1 signaling complex. Immunity 4, 387–396 (1996).

    CAS  Article  Google Scholar 

  18. 18

    Lewis, J. et al. Disruption of hsp90 function results in degradation of the death domain kinase, receptor-interacting protein (RIP), and blockage of tumor necrosis factor–induced nuclear factor-κB activation. J. Biol. Chem. 275, 10519–10526 (2000).

    CAS  Article  Google Scholar 

  19. 19

    Zhang, S. Q., Kovalenko, A., Cantarella, G. & Wallach, D. Recruitment of the IKK signalosome to the p55 TNF receptor: RIP and A20 bind to NEMO (IKKγ) upon receptor stimulation. Immunity 12, 301–311 (2000).

    CAS  Article  Google Scholar 

  20. 20

    Kroemer, G. & Reed, J. C. Mitochondrial control of cell death. Nature Med. 6, 513–519 (2000).

    CAS  Article  Google Scholar 

  21. 21

    Li, H., Zhu, H., Xu, C. J. & Yuan, J. Cleavage of BID by caspase 8 mediates the mitochondrial damage in the Fas pathway of apoptosis. Cell 94, 491–501 (1998).

    CAS  Article  Google Scholar 

  22. 22

    Luo, X., Budihardjo, I., Zou, H., Slaughter, C. & Wang, X. Bid, a Bcl2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. Cell 94, 481–490 (1998).

    CAS  Article  Google Scholar 

  23. 23

    Vercammen, D. et al. Inhibition of caspases increases the sensitivity of L929 cells to necrosis mediated by tumor necrosis factor. J. Exp. Med. 187, 1477–1485 (1998).

    CAS  Article  Google Scholar 

  24. 24

    Pimentel-Muinos, F. X. & Seed, B. Regulated commitment of TNF receptor signaling: a molecular switch for death or activation. Immunity 11, 783–793 (1999).

    CAS  Article  Google Scholar 

  25. 25

    Bodmer, J. L. et al. TRAIL receptor-2 signals apoptosis through FADD and caspase-8. Nature Cell Biol. 2, 241–243 (2000).

    CAS  Article  Google Scholar 

  26. 26

    Kischkel, F. C. et al. Apo2L/TRAIL-dependent recruitment of endogenous FADD and caspase-8 to death receptors 4 and 5. Immunity 12, 611–620 (2000).

    CAS  Article  Google Scholar 

  27. 27

    Sprick, M. R. et al. FADD/MORT1 and caspase-8 are recruited to TRAIL receptors 1 and 2 and are essential for apoptosis mediated by TRAIL receptor 2. Immunity 12, 599–609 (2000).

    CAS  Article  Google Scholar 

  28. 28

    Khwaja, A. & Tatton, L. Resistance to the cytotoxic effects of tumor necrosis factor alpha can be overcome by inhibition of a FADD/caspase-dependent signaling pathway. J. Biol. Chem. 274, 36817–36823 (1999).

    CAS  Article  Google Scholar 

  29. 29

    Boone, E. et al. Structure/Function Analysis of p55 Tumor Necrosis Factor Receptor and Fas-Associated Death Domain: Effect on Necrosis in L929sA Cells. J. Biol. Chem. (in the press2000).

  30. 30

    Luschen, S., Ussat, S., Scherer, G., Kabelitz, D. & Adam-Klages, S. Sensitization to death receptor cytotoxicity by inhibition of fas- associated death domain protein (FADD)/caspase signaling. Requirement of cell cycle progression. J. Biol. Chem. 275, 24670–24678 (2000).

    CAS  Article  Google Scholar 

  31. 31

    Walsh, C. M. et al. A role for FADD in T cell activation and development. Immunity 8, 439–449 (1998).

    CAS  Article  Google Scholar 

  32. 32

    Watanabe, F. R., Brannan, C. I., Copeland, N. G., Jenkins, N. A. & Nagata, S. Lymphoproliferation disorder in mice explained by defects in Fas antigen that mediates apoptosis. Nature 356, 314–317 (1992).

    Article  Google Scholar 

  33. 33

    Gallucci, S., Lolkema, M. & Matzinger, P. Natural adjuvants: endogenous activators of dendritic cells. Nature Med. 5, 1249–1255 (1999).

    CAS  Article  Google Scholar 

  34. 34

    Baker, B., Zambryski, P., Staskawicz, B. & Dinesh-Kumar, S. P. Signaling in plant-microbe interactions. Science 276, 726–733 (1997).

    CAS  Article  Google Scholar 

  35. 35

    Berndt, C., Mopps, B., Angermuller, S., Gierschik, P. & Krammer, P. H. CXCR4 and CD4 mediate a rapid CD95-independent cell death in CD4(+) T cells. Proc. Natl Acad. Sci. USA 95, 12556–12561 (1998).

    CAS  Article  Google Scholar 

  36. 36

    Drenou, B. et al. A caspase-independent pathway of MHC class II antigen-mediated apoptosis of human B lymphocytes. J. Immunol. 163, 4115–4124 (1999).

    CAS  PubMed  Google Scholar 

  37. 37

    Kitanaka, C. & Kuchino, Y. Caspase-independent programmed cell death with necrotic morphology. Cell Death Differ. 6, 508–515 (1999).

    CAS  Article  Google Scholar 

Download references


We are grateful to A. Strasser for the gift of the CEM-FADD/DN and CEM-CrmA clones and J. Blenis for the FADD and caspase-8 deficient Jurkat cells. We thank K. Burns and L. French for helpful comments and C. Mattmann for technical assistance.

Author information



Corresponding author

Correspondence to Jürg Tschopp.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Holler, N., Zaru, R., Micheau, O. et al. Fas triggers an alternative, caspase-8–independent cell death pathway using the kinase RIP as effector molecule. Nat Immunol 1, 489–495 (2000).

Download citation

Further reading


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