Abstract
Pattern-recognition receptors (PRRs) elicit antiviral immune responses to human immunodeficiency virus type 1 (HIV-1). Here we show that HIV-1 required signaling by the PRRs Toll-like receptor 8 (TLR8) and DC-SIGN for replication in dendritic cells (DCs). HIV-1 activated the transcription factor NF-κB through TLR8 to initiate the transcription of integrated provirus by RNA polymerase II (RNAPII). However, DC-SIGN signaling was required for the generation of full-length viral transcripts. Binding of the HIV-1 envelope glycoprotein gp120 to DC-SIGN induced kinase Raf-1–dependent phosphorylation of the NF-κB subunit p65 at Ser276, which recruited the transcription-elongation factor pTEF-b to nascent transcripts. Transcription elongation and generation of full-length viral transcripts was dependent on pTEF-b-mediated phosphorylation of RNAPII at Ser2. Inhibition of either pathway abrogated replication and prevented HIV-1 transmission. Thus, HIV-1 subverts crucial components of the immune system for replication that might be targeted to prevent infection and dissemination.
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Acknowledgements
We thank M. Oudhoff (Academic Centre for Dentistry Amsterdam) for C. albicans; P. Gallay (The Scripps Research Institute) and C. Aiken (Vanderbilt University School of Medicine) for HIV-1 proviral plasmids; K. Kristiansen (University of Southern Denmark) for the GST-p65 expression plasmid; J. Belisle (Colorado State University; National Institute of Allergy and Infectious Diseases contract HHSN266200400091C) for M. tuberculosis; and J. Kappes and X. Wu (Tranzyme) for TZM-bl cells (obtained through the National Institutes of Health AIDS Research and Reference Reagent Program). Supported by the Netherlands Organisation for Scientific Research (NWO 917-46-367 to M.L. and NWO 912-04-025 to J.d.D.), the AIDS Foundation (2007036 to M.v.d.V.), Dutch Burns Foundation (08.109 to L.M.v.d.B) and the Dutch Asthma Foundation (3.2.03.39 to S.I.G.).
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S.I.G. designed, executed and interpreted most experiments and prepared the manuscript; M.v.d.V. executed HIV-1 stimulation, infection and transmission experiments, isolated myeloid DCs and helped with the nuclear extract preparations; L.M.v.d.B. isolated dermal DCs; J.d.D. helped with flow cytometry; M.L. helped with GST-p65 purification and setting up the GST enzyme-linked immunosorbent assay (ELISA); and T.B.H.G. supervised all aspects of this study, including study design, execution and interpretation, and manuscript preparation.
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Gringhuis, S., van der Vlist, M., van den Berg, L. et al. HIV-1 exploits innate signaling by TLR8 and DC-SIGN for productive infection of dendritic cells. Nat Immunol 11, 419–426 (2010). https://doi.org/10.1038/ni.1858
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DOI: https://doi.org/10.1038/ni.1858
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