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Cell death by pyroptosis drives CD4 T-cell depletion in HIV-1 infection

Nature volume 505, pages 509514 (23 January 2014) | Download Citation

  • A Corrigendum to this article was published on 22 March 2017


The pathway causing CD4 T-cell death in HIV-infected hosts remains poorly understood although apoptosis has been proposed as a key mechanism. We now show that caspase-3-mediated apoptosis accounts for the death of only a small fraction of CD4 T cells corresponding to those that are both activated and productively infected. The remaining over 95% of quiescent lymphoid CD4 T cells die by caspase-1-mediated pyroptosis triggered by abortive viral infection. Pyroptosis corresponds to an intensely inflammatory form of programmed cell death in which cytoplasmic contents and pro-inflammatory cytokines, including IL-1β, are released. This death pathway thus links the two signature events in HIV infection—CD4 T-cell depletion and chronic inflammation—and creates a pathogenic vicious cycle in which dying CD4 T cells release inflammatory signals that attract more cells to die. This cycle can be broken by caspase 1 inhibitors shown to be safe in humans, raising the possibility of a new class of ‘anti-AIDS’ therapeutics targeting the host rather than the virus.

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  1. 1.

    Roadblocks in HIV research: five questions. Nature Med. 15, 855–859 (2009)

  2. 2.

    , & Analysis of apoptosis in lymph nodes of HIV-infected persons. Intensity of apoptosis correlates with the general state of activation of the lymphoid tissue and not with stage of disease or viral burden. J. Immunol. 154, 5555–5566 (1995)

  3. 3.

    et al. Apoptosis occurs predominantly in bystander cells and not in productively infected cells of HIV- and SIV-infected lymph nodes. Nature Med. 1, 129–134 (1995)

  4. 4.

    , , & Apoptotic peptides derived from HIV-1 Nef induce lymphocyte depletion in mice. Ethnic. Dis. 18, S2–30–37 (2008)

  5. 5.

    et al. Correlates of apoptosis of CD4+ and CD8+ T cells in tonsillar tissue in HIV type 1 infection. AIDS Res. Hum. Retroviruses 14, 1635–1643 (1998)

  6. 6.

    et al. Programmed cell death in peripheral lymphocytes from HIV-infected persons: increased susceptibility to apoptosis of CD4 and CD8 T cells correlates with lymphocyte activation and with disease progression. J. Immunol. 156, 3509–3520 (1996)

  7. 7.

    et al. In vivo evolution of human immunodeficiency virus type 1 toward increased pathogenicity through CXCR4-mediated killing of uninfected CD4 T cells. J. Virol. 77, 5846–5854 (2003)

  8. 8.

    , & Human immunodeficiency virus type 1 induces apoptosis in CD4+ but not in CD8+ T cells in ex vivo-infected human lymphoid tissue. J. Virol. 74, 8077–8084 (2000)

  9. 9.

    & Manipulation of host cell death pathways during microbial infections. Cell Host Microbe 8, 44–54 (2010)

  10. 10.

    et al. HIV-1 causes CD4 cell death through DNA-dependent protein kinase during viral integration. Nature 498, 376–379 (2013)

  11. 11.

    et al. Abortive HIV infection mediates CD4 T cell depletion and inflammation in human lymphoid tissue. Cell 143, 789–801 (2010)

  12. 12.

    et al. HIV-1 actively replicates in naive CD4+ T cells residing within human lymphoid tissues. Immunity 15, 671–682 (2001)

  13. 13.

    , , & Infection of human tonsil histocultures: a model for HIV pathogenesis. Nature Med. 1, 1320–1322 (1995)

  14. 14.

    & Apoptosis: the importance of being eaten. Cell Death Differ. 5, 563–568 (1998)

  15. 15.

    & Apoptosis, pyroptosis, and necrosis: mechanistic description of dead and dying eukaryotic cells. Infect. Immun. 73, 1907–1916 (2005)

  16. 16.

    , , & Dynamics of HIV-1 recombination in its natural target cells. Proc. Natl Acad. Sci. USA 101, 4204–4209 (2004)

  17. 17.

    , , & Activation of caspases measured in situ by binding of fluorochrome-labeled inhibitors of caspases (FLICA): correlation with DNA fragmentation. Exp. Cell Res. 259, 308–313 (2000)

  18. 18.

    et al. An infectious molecular clone of an unusual macrophage-tropic and highly cytopathic strain of human immunodeficiency virus type 1. J. Virol. 66, 7517–7521 (1992)

  19. 19.

    et al. Cryopyrin activates the inflammasome in response to toxins and ATP. Nature 440, 228–232 (2006)

  20. 20.

    & The inflammasomes. Cell 140, 821–832 (2010)

  21. 21.

    et al. Differential requirement for the activation of the inflammasome for processing and release of IL-1β in monocytes and macrophages. Blood 113, 2324–2335 (2009)

  22. 22.

    , & ATP treatment of human monocytes promotes caspase-1 maturation and externalization. J. Biol. Chem. 274, 36944–36951 (1999)

  23. 23.

    & Interleukin-1β maturation and release in response to ATP and nigericin. Evidence that potassium depletion mediated by these agents is a necessary and common feature of their activity. J. Biol. Chem. 269, 15195–15203 (1994)

  24. 24.

    et al. IL-1β maturation: evidence that mature cytokine formation can be induced specifically by nigericin. J. Immunol. 149, 1294–1303 (1992)

  25. 25.

    , , & The CCR5 and CXCR4 coreceptors–central to understanding the transmission and pathogenesis of human immunodeficiency virus type 1 infection. AIDS Res. Hum. Retroviruses 20, 111–126 (2004)

  26. 26.

    et al. R5 human immunodeficiency virus type 1 (HIV-1) replicates more efficiently in primary CD4+ T-cell cultures than X4 HIV-1. J. Virol. 78, 9164–9173 (2004)

  27. 27.

    & CCR5- and CXCR4-tropic HIV-1 are equally cytopathic for their T-cell targets in human lymphoid tissue. Nature Med. 5, 344–346 (1999)

  28. 28.

    , , & Preferential cytolysis of peripheral memory CD4+ T cells by in vitro X4-tropic human immunodeficiency virus type 1 infection before the completion of reverse transcription. J. Virol. 82, 9154–9163 (2008)

  29. 29.

    & Dynamics of T lymphocyte responses: intermediates, effectors, and memory cells. Science 290, 92–97 (2000)

  30. 30.

    Immunological memory. Adv. Immunol. 53, 217–265 (1993)

  31. 31.

    , , , & Two subsets of memory T lymphocytes with distinct homing potentials and effector functions. Nature 401, 708–712 (1999)

  32. 32.

    , & Pyroptosis: host cell death and inflammation. Nature Rev. Microbiol. 7, 99–109 (2009)

  33. 33.

    & A quick and simple method for the quantitation of lactate dehydrogenase release in measurements of cellular cytotoxicity and tumor necrosis factor (TNF) activity. J. Immunol. Methods 115, 61–69 (1988)

  34. 34.

    et al. Cre-lox-regulated conditional RNA interference from transgenes. Proc. Natl Acad. Sci. USA 101, 10380–10385 (2004)

  35. 35.

    et al. SAMHD1 restricts HIV-1 infection in resting CD4+ T cells. Nature Med. 18, 1682–1687 (2012)

  36. 36.

    et al. The CXCR4-tropic human immunodeficiency virus envelope promotes more-efficient gene delivery to resting CD4+ T cells than the vesicular stomatitis virus glycoprotein G envelope. J. Virol. 83, 8153–8162 (2009)

  37. 37.

    , , , & A small molecule inhibitor of Caspase-1. in Probe Reports from the NIH Molecular Libraries Program (Bethesda Maryland, 2010)

  38. 38.

    et al. A highly potent and selective caspase-1 inhibitor that utilizes a key 3-cyanopropanoic acid moiety. ChemMedChem 5, 730–738 (2010)

  39. 39.

    , , , & ICE/Caspase-1 inhibitors as novel anti-inflammatory drugs. Expert Opin. Investig. Drugs 10, 1207–1209 (2001)

  40. 40.

    et al. IL-converting enzyme/caspase-1 inhibitor VX-765 blocks the hypersensitive response to an inflammatory stimulus in monocytes from familial cold autoinflammatory syndrome patients. J. Immunol. 175, 2630–2634 (2005)

  41. 41.

    et al. Interleukin-1β biosynthesis inhibition reduces acute seizures and drug resistant chronic epileptic activity in mice. Neurotherapeutics 8, 304–315 (2011)

  42. 42.

    et al. ICE/caspase 1 inhibitors and IL-1β receptor antagonists as potential therapeutics in epilepsy. Current Opinion in Investigational Drugs 11, 43–50 (2010)

  43. 43.

    , & CD4+ T cell depletion in human immunodeficiency virus (HIV) infection: role of apoptosis. Viruses 3, 586–612 (2011)

  44. 44.

    et al. IFI16 DNA sensor is required for death of lymphoid CD4 T cells abortively infected with HIV. Science (19 December 2013)

  45. 45.

    et al. HIV-1 induced activation of CD4+ T cells creates new targets for HIV-1 infection in human lymphoid tissue ex vivo. Blood 111, 699–704 (2008)

  46. 46.

    et al. Cumulative mechanisms of lymphoid tissue fibrosis and T cell depletion in HIV-1 and SIV infections. J. Clin. Invest. 121, 998–1008 (2011)

  47. 47.

    HIV infection, inflammation, immunosenescence, and aging. Annu. Rev. Med. 62, 141–155 (2011)

  48. 48.

    , , & Nef enhances human immunodeficiency virus type 1 infectivity and replication independently of viral coreceptor tropism. J. Virol. 76, 8455–8459 (2002)

  49. 49.

    et al. With a little help from a friend: increasing HIV transduction of monocyte-derived dendritic cells with virion-like particles of SIV(MAC). Gene Ther. 13, 991–994 (2006)

  50. 50.

    et al. SAMHD1 is the dendritic- and myeloid-cell-specific HIV-1 restriction factor counteracted by Vpx. Nature 474, 654–657 (2011)

  51. 51.

    , , , & Endogenous APOBEC3B restricts LINE-1 retrotransposition in transformed cells and human embryonic stem cells. J. Biol. Chem. 286, 36427–36437 (2011)

  52. 52.

    et al. HIV-1 pathogenesis differs in rectosigmoid and tonsillar tissues infected ex vivo with CCR5- and CXCR4-tropic HIV-1. AIDS 21, 1263–1272 (2007)

  53. 53.

    , , & Biochemical transfer of single-copy eucaryotic genes using total cellular DNA as donor. Cell. 14, 725–731 (1978)

  54. 54.

    et al. A dual-tropic primary HIV-1 isolate that uses fusin and the beta-chemokine receptors CKR-5, CKR-3, and CKR-2b as fusion cofactors. Cell 85, 1149–1158 (1996)

  55. 55.

    , , & 11-color, 13-parameter flow cytometry: identification of human naive T cells by phenotype, function, and T-cell receptor diversity. Nature Med. 7, 245–248 (2001)

  56. 56.

    et al. CD4+ T cell depletion during all stages of HIV disease occurs predominantly in the gastrointestinal tract. J. Exp. Med. 200, 749–759 (2004)

  57. 57.

    , , , & The HIV coreceptors CXCR4 and CCR5 are differentially expressed and regulated on human T lymphocytes. Proc. Natl Acad. Sci. USA 94, 1925–1930 (1997)

  58. 58.

    et al. Segregation of R5 and X4 HIV-1 variants to memory T cell subsets differentially expressing CD62L in ex vivo infected human lymphoid tissue. AIDS 16, 1245–1249 (2002)

  59. 59.

    , , , & CXCR4 utilization is sufficient to trigger CD4+ T cell depletion in HIV-1-infected human lymphoid tissue. Proc. Natl Acad. Sci. USA 96, 663–668 (1999)

  60. 60.

    et al. Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates programmed necrosis and virus-induced inflammation. Cell 137, 1112–1123 (2009)

  61. 61.

    Innate antiviral response: role in HIV-1 infection. Viruses 3, 1179–1203 (2011)

  62. 62.

    Antiviral actions of interferons. Clinical Microbiology Reviews 14, 778–809 (2001)

  63. 63.

    & The inflammasomes. PLoS Pathog. 5, e1000510 (2009)

  64. 64.

    & The cytokine release inhibitory drug CRID3 targets ASC oligomerisation in the NLRP3 and AIM2 inflammasomes. PLoS ONE 6, e29539 (2011)

  65. 65.

    et al. Anti-inflammatory compounds parthenolide and Bay 11-7082 are direct inhibitors of the inflammasome. J. Biol. Chem. 285, 9792–9802 (2010)

  66. 66.

    et al. Glyburide inhibits the Cryopyrin/Nalp3 inflammasome. J. Cell Biol. 187, 61–70 (2009)

  67. 67.

    et al. Characterization of human blood dendritic cell subsets. Blood 100, 4512–4520 (2002)

  68. 68.

    , , , & The dendritic cell lineage: ontogeny and function of dendritic cells and their subsets in the steady state and the inflamed setting. Annu. Rev. Immunol. 31, 563–604 (2013)

  69. 69.

    et al. SAMHD1 restricts HIV-1 reverse transcription in quiescent CD4+ T-cells. Retrovirology 9, 87 (2012)

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We thank D. N. Levy for the NLENG1 plasmid; L. A. J. O’Neill for CRID3 and parthenolide; R. Collman for the HIV-1 89.6 clone; and Vertex Pharmaceuticals for the VX-765 and VRT-043198 compounds. HIV-infected lymph node tissue was obtained from the SCOPE cohort at HIV/AIDS clinic of the San Francisco General Hospital (SFGH) Positive Health Program, with the help of R. Hoh, and M. Kerbelski. We thank W. Schecter for surgical removal of the lymph nodes from HIV-infected subjects. We thank L. Napolitano and Y. Lie from Monogram Biosciences for performing Trofile assays to determine HIV co-receptor tropism in samples of HIV-infected volunteers. The following reagents were obtained through the AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH: AMD3100, efavirenz and raltegravir. We thank C. Miller, director of the Gladstone Histology Core for performing the immunostaining assays and M. Cavrois, M. Gesner, and J. Tawney for assistance with flow cytometry. We also thank G. Howard and A. L. Lucido for editorial assistance; J. C. W. Carroll, G. Maki, and T. Roberts for graphics arts; and R. Givens and S. Cammack for administrative assistance. Special thanks to N. Roan for comments on the manuscript and to J. Neidleman for stimulating discussions and technical advice. We thank the NIH/NIAID for funding (R21AI102782, 1DP1036502, U19 AI0961133). Funding was also provided by the UCSF/Robert John Sabo Trust Award (G.D.) and A.P. Giannini Foundation Postdoctoral Research Fellowship (K.M.M.). We also acknowledge support from NIH P30 AI027763 (UCSF-GIVI Center for AIDS Research) for support to S.S. and Z.Y., and for Immunology Core services.

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Author notes

    • Gilad Doitsh
    • , Nicole L. K. Galloway
    •  & Xin Geng

    These authors contributed equally to this work.


  1. Gladstone Institute of Virology and Immunology, 1650 Owens Street, San Francisco, California 94158, USA

    • Gilad Doitsh
    • , Nicole L. K. Galloway
    • , Xin Geng
    • , Zhiyuan Yang
    • , Kathryn M. Monroe
    • , Orlando Zepeda
    • , Stefanie Sowinski
    • , Isa Muñoz-Arias
    •  & Warner C. Greene
  2. Department of Medicine, University of California, San Francisco, 505 Parnassus Avenue, San Francisco, California 94143, USA

    • Peter W. Hunt
    • , Hiroyu Hatano
    •  & Warner C. Greene
  3. Department of Microbiology and Immunology, University of California, San Francisco, 505 Parnassus Avenue, San Francisco, California 94143, USA

    • Warner C. Greene


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G.D. identified the involvement of caspase 1 and pyroptosis in lymphoid CD4 T-cell death by HIV-1, developed and designed most of the studies, collected the data and wrote the manuscript; N.L.K.G. performed IL-1β protein assays and examined VX-765 in HIV-infected tonsils; X.G. performed FLICA and shRNA analyses in HLACs; Z.Y. analysed caspase cleavage in HIV-infected cultures; K.M.M. examined caspase inhibitors and LDH release assays; O.Z. tested caspase inhibitors, type-I IFN, and pro-IL-1β expression; P.W.H. and H.H. provided HIV-infected lymphoid node from surgeries of SCOPE cohort patients at HIV/AIDS clinic of the San Francisco General Hospital (SFGH); I.M.-A. provided reagents and tissues; S.S. coordinated lymph node biopsies; W.C.G. supervised all of these studies and participated in the preparation of the final manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Warner C. Greene.

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