SAMHD1 restricts HIV-1 infection in resting CD4+ T cells

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Unlike activated CD4+ T cells, resting CD4+ T cells are highly resistant to productive HIV-1 infection1,2,3,4,5,6,7,8. Early after HIV-1 entry, a major block limits reverse transcription of incoming viral genomes. Here we show that the deoxynucleoside triphosphate triphosphohydrolase SAMHD1 prevents reverse transcription of HIV-1 RNA in resting CD4+ T cells. SAMHD1 is abundantly expressed in resting CD4+ T cells circulating in peripheral blood and residing in lymphoid organs. The early restriction to infection in unstimulated CD4+ T cells is overcome by HIV-1 or HIV-2 virions into which viral Vpx is artificially or naturally packaged, respectively, or by addition of exogenous deoxynucleosides. Vpx-mediated proteasomal degradation of SAMHD1 and elevation of intracellular deoxynucleotide pools precede successful infection by Vpx-carrying HIV. Resting CD4+ T cells from healthy donors following SAMHD1 silencing or from a patient with Aicardi-Goutières syndrome homozygous for a nonsense mutation in SAMHD1 were permissive for HIV-1 infection. Thus, SAMHD1 imposes an effective restriction to HIV-1 infection in the large pool of noncycling CD4+ T cells in vivo. Bypassing SAMHD1 was insufficient for the release of viral progeny, implicating other barriers at later stages of HIV replication. Together, these findings may unveil new ways to interfere with the immune evasion and T cell immunopathology of pandemic HIV-1.

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Figure 1: Vpx overcomes a restriction to HIV-1 and HIV-2 infection in resting human CD4+ T cells.
Figure 2: SAMHD1 is expressed in resting CD4+ T cells and depleted by Vpx.
Figure 3: Susceptibility of resting CD4+ T cells to Vpx-carrying HIV-1 is paralleled by proteasomal degradation of SAMHD1 and increased dNTP levels.
Figure 4: Silencing of SAMHD1 by RNA interference or a homozygous nonsense mutation renders resting CD4+ T cells permissive for HIV-1 infection.


  1. 1

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

  2. 2

    Ganesh, L. et al. The gene product Murr1 restricts HIV-1 replication in resting CD4+ lymphocytes. Nature 426, 853–857 (2003).

  3. 3

    Korin, Y.D. & Zack, J.A. Progression to the G1b phase of the cell cycle is required for completion of human immunodeficiency virus type 1 reverse transcription in T cells. J. Virol. 72, 3161–3168 (1998).

  4. 4

    Pierson, T.C. et al. Molecular characterization of preintegration latency in human immunodeficiency virus type 1 infection. J. Virol. 76, 8518–8531 (2002).

  5. 5

    Stevenson, M., Stanwick, T.L., Dempsey, M.P. & Lamonica, C.A. HIV-1 replication is controlled at the level of T cell activation and proviral integration. EMBO J. 9, 1551–1560 (1990).

  6. 6

    Zack, J.A. et al. HIV-1 entry into quiescent primary lymphocytes: molecular analysis reveals a labile, latent viral structure. Cell 61, 213–222 (1990).

  7. 7

    Dai, J. et al. Human immunodeficiency virus integrates directly into naive resting CD4+ T cells but enters naive cells less efficiently than memory cells. J. Virol. 83, 4528–4537 (2009).

  8. 8

    Plesa, G. et al. Addition of deoxynucleosides enhances human immunodeficiency virus type 1 integration and 2LTR formation in resting CD4+ T cells. J. Virol. 81, 13938–13942 (2007).

  9. 9

    Yoder, A. et al. HIV envelope-CXCR4 signaling activates cofilin to overcome cortical actin restriction in resting CD4 T cells. Cell 134, 782–792 (2008).

  10. 10

    Swingler, S. et al. HIV-1 Nef intersects the macrophage CD40L signalling pathway to promote resting-cell infection. Nature 424, 213–219 (2003).

  11. 11

    Bergamaschi, A. et al. The human immunodeficiency virus type 2 Vpx protein usurps the CUL4A-DDB1 DCAF1 ubiquitin ligase to overcome a postentry block in macrophage infection. J. Virol. 83, 4854–4860 (2009).

  12. 12

    Berger, G., Goujon, C., Darlix, J.L. & Cimarelli, A. SIVMAC Vpx improves the transduction of dendritic cells with nonintegrative HIV-1–derived vectors. Gene Ther. 16, 159–163 (2009).

  13. 13

    Goujon, C. et al. Characterization of simian immunodeficiency virus SIVSM/human immunodeficiency virus type 2 Vpx function in human myeloid cells. J. Virol. 82, 12335–12345 (2008).

  14. 14

    Pertel, T., Reinhard, C. & Luban, J. Vpx rescues HIV-1 transduction of dendritic cells from the antiviral state established by type 1 interferon. Retrovirology 8, 49 (2011).

  15. 15

    Sharova, N. et al. Primate lentiviral Vpx commandeers DDB1 to counteract a macrophage restriction. PLoS Pathog. 4, e1000057 (2008).

  16. 16

    Srivastava, S. et al. Lentiviral Vpx accessory factor targets VprBP/DCAF1 substrate adaptor for cullin 4 E3 ubiquitin ligase to enable macrophage infection. PLoS Pathog. 4, e1000059 (2008).

  17. 17

    Berger, A. et al. SAMHD1-deficient CD14+ cells from individuals with Aicardi-Goutieres syndrome are highly susceptible to HIV-1 infection. PLoS Pathog. 7, e1002425 (2011).

  18. 18

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

  19. 19

    Hrecka, K. et al. Vpx relieves inhibition of HIV-1 infection of macrophages mediated by the SAMHD1 protein. Nature 474, 658–661 (2011).

  20. 20

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

  21. 21

    Goldstone, D.C. et al. HIV-1 restriction factor SAMHD1 is a deoxynucleoside triphosphate triphosphohydrolase. Nature 480, 379–382 (2011).

  22. 22

    Lahouassa, H. et al. SAMHD1 restricts the replication of human immunodeficiency virus type 1 by depleting the intracellular pool of deoxynucleoside triphosphates. Nat. Immunol. 13, 223–228 (2012).

  23. 23

    Powell, R.D., Holland, P.J., Hollis, T. & Perrino, F.W. Aicardi-Goutieres syndrome gene and HIV-1 restriction factor SAMHD1 is a dGTP-regulated deoxynucleotide triphosphohydrolase. J. Biol. Chem. 286, 43596–43600 (2011).

  24. 24

    Diamond, T.L. et al. Macrophage tropism of HIV-1 depends on efficient cellular dNTP utilization by reverse transcriptase. J. Biol. Chem. 279, 51545–51553 (2004).

  25. 25

    Wabnitz, G.H. et al. Sustained LFA-1 cluster formation in the immune synapse requires the combined activities of L-plastin and calmodulin. Eur. J. Immunol. 40, 2437–2449 (2010).

  26. 26

    Santoni de Sio, F.R. & Trono, D. APOBEC3G-depleted resting CD4+ T cells remain refractory to HIV1 infection. PLoS ONE 4, e6571 (2009).

  27. 27

    Thiele, H. et al. Cerebral arterial stenoses and stroke: novel features of Aicardi-Goutieres syndrome caused by the Arg164X mutation in SAMHD1 are associated with altered cytokine expression. Hum. Mutat. 31, E1836–E1850 (2010).

  28. 28

    Lim, E.S. et al. The ability of primate lentiviruses to degrade the monocyte restriction factor SAMHD1 preceded the birth of the viral accessory protein Vpx. Cell Host Microbe 11, 194–204 (2012).

  29. 29

    Laguette, N. et al. Evolutionary and functional analyses of the interaction between the myeloid restriction factor SAMHD1 and the lentiviral Vpx protein. Cell Host Microbe 11, 205–217 (2012).

  30. 30

    Manel, N. et al. A cryptic sensor for HIV-1 activates antiviral innate immunity in dendritic cells. Nature 467, 214–217 (2010).

  31. 31

    Manel, N. & Littman, D.R. Hiding in plain sight: how HIV evades innate immune responses. Cell 147, 271–274 (2011).

  32. 32

    Keppler, O.T. et al. Susceptibility of rat-derived cells to replication by human immunodeficiency virus type 1. J. Virol. 75, 8063–8073 (2001).

  33. 33

    Haller, C. et al. The HIV-1 pathogenicity factor Nef interferes with maturation of stimulatory T-lymphocyte contacts by modulation of N-Wasp activity. J. Biol. Chem. 281, 19618–19630 (2006).

  34. 34

    Goffinet, C. et al. HIV-1 antagonism of CD317 is species specific and involves Vpu-mediated proteasomal degradation of the restriction factor. Cell Host Microbe 5, 285–297 (2009).

  35. 35

    Goffinet, C., Allespach, I. & Keppler, O.T. HIV-susceptible transgenic rats allow rapid preclinical testing of antiviral compounds targeting virus entry or reverse transcription. Proc. Natl. Acad. Sci. USA 104, 1015–1020 (2007).

  36. 36

    Sunseri, N., O'Brien, M., Bhardwaj, N. & Landau, N.R. Human immunodeficiency virus type 1 modified to package simian immunodeficiency virus Vpx efficiently infects macrophages and dendritic cells. J. Virol. 85, 6263–6274 (2011).

  37. 37

    Gramberg, T., Sunseri, N. & Landau, N.R. Evidence for an activation domain at the amino terminus of simian immunodeficiency virus Vpx. J. Virol. 84, 1387–1396 (2010).

  38. 38

    Berger, A. et al. Interaction of Vpx and apolipoprotein B mRNA-editing catalytic polypeptide 3 family member A (APOBEC3A) correlates with efficient lentivirus infection of monocytes. J. Biol. Chem. 285, 12248–12254 (2010).

  39. 39

    Keppler, O.T., Tibroni, N., Venzke, S., Rauch, S. & Fackler, O.T. Modulation of specific surface receptors and activation sensitization in primary resting CD4+ T lymphocytes by the Nef protein of HIV-1. J. Leukoc. Biol. 79, 616–627 (2006).

  40. 40

    Geuenich, S. et al. Aqueous extracts from peppermint, sage and lemon balm leaves display potent anti-HIV-1 activity by increasing the virion density. Retrovirology 5, 27 (2008).

  41. 41

    Tervo, H.M., Goffinet, C. & Keppler, O.T. Mouse T-cells restrict replication of human immunodeficiency virus at the level of integration. Retrovirology 5, 58 (2008).

  42. 42

    Keppler, O.T. et al. Rodent cells support key functions of the human immunodeficiency virus type 1 pathogenicity factor Nef. J. Virol. 79, 1655–1665 (2005).

  43. 43

    Ruggieri, A. et al. Human endogenous retrovirus HERV-K(HML-2) encodes a stable signal peptide with biological properties distinct from Rec. Retrovirology 6, 17 (2009).

  44. 44

    Erikson, E. et al. In vivo expression profile of the antiviral restriction factor and tumor-targeting antigen CD317/BST-2/HM1.24/tetherin in humans. Proc. Natl. Acad. Sci. USA 108, 13688–13693 (2011).

  45. 45

    Swiggard, W.J. et al. Human immunodeficiency virus type 1 can establish latent infection in resting CD4+ T cells in the absence of activating stimuli. J. Virol. 79, 14179–14188 (2005).

  46. 46

    Goffinet, C., Schmidt, S., Kern, C., Oberbremer, L. & Keppler, O.T. Endogenous CD317/Tetherin limits replication of HIV-1 and murine leukemia virus in rodent cells and is resistant to antagonists from primate viruses. J. Virol. 84, 11374–11384 (2010).

  47. 47

    Verhoeyen, E. et al. IL-7 surface-engineered lentiviral vectors promote survival and efficient gene transfer in resting primary T lymphocytes. Blood 101, 2167–2174 (2003).

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We thank M. Emerman (Fred Hutchinson Cancer Research Center, Seattle, USA; for pROD9, pROD9-ΔEnv-GFP, pROD9-ΔEnv-delVpx-GFP), M. Fujita (Research Institute for Drug Discovery, Kumamoto University, Japan; for pEF-FVpxHIV-2GH1), H.-G. Kräusslich (Department of Infectious Diseases, Virology, University of Heidelberg, Germany; for AMD3100), B. Müller (Department of Infectious Diseases, Virology, University of Heidelberg, Germany; for pDisplay GFP and sheep anti–HIV-1 p24 antibody) and J. Münch (Institute of Molecular Virology, University Hospital Ulm, Germany; for HIV-1 GFP) for reagents. We thank T. Adam, A. Imle, P. Klein, S. Kutscheidt, A. Ruggieri and the Nikon Imaging Center at University of Heidelberg for technical assistance, and G. Howard for editorial assistance. This work was in part funded by the Deutsche Forschungsgemeinschaft (O.T.K., grant KE742/4-1), SFB 938/Z2 (F.L.) and the US National Institutes of Health (GM1041981 and AI049781 to B.K.; and F31 GM095190 to W.D.). O.T.F. and O.T.K. are members of the CellNetworks Cluster of Excellence EXC81 and the German Centre for Infection Research, University of Heidelberg. H.-M.B. is recipient of a fellowship of the Medical Faculty of the University of Heidelberg.

Author information

O.T.K. and O.T.F. conceived of the study, designed experiments and wrote the manuscript. H-M.B., X.P., E.E., S. Schmidt, W.D., M.B., K.S., B.K., S.P. and R.K. designed and conducted experiments and discussed and interpreted the data with O.T.F. and O.T.K. I.A., E.F., G.W., T.G. and N.R.L. provided reagents and expertise. S. Sertel, F.R. and F.L. provided tissue samples and expertise and interpreted data.

Correspondence to Oliver T Fackler or Oliver T Keppler.

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Supplementary Figures 1–12 (PDF 1261 kb)

Supplementary Video 1

Subcellular localization of SAMHD1 in resting CD4+ T cells. 360° rotating movie of the 3D reconstruction of deconvolution confocal images of the cell shown in Figure 2e, with SAMHD1 depicted in green and lamin A shown in red. In the second half of the movie, the lamin A stain of the nuclear envelope is rendered transparent to unmask the nuclear portion of SAMHD1. (AVI 6095 kb)

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Baldauf, H., Pan, X., Erikson, E. et al. SAMHD1 restricts HIV-1 infection in resting CD4+ T cells. Nat Med 18, 1682–1688 (2012) doi:10.1038/nm.2964

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