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

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Abstract

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.

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Acknowledgements

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 Text and Figures

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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