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TLR-mediated induction of negative regulatory ligands on dendritic cells

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Abstract

Dendritic cells (DCs) shape T-cell response patterns and determine early, intermediate, and late outcomes of immune recognition events. They either facilitate immunostimulation or induce tolerance, possibly determined by initial DC activation signals, such as binding Toll-like receptor (TLR) ligands. Here, we report that DC stimulation through the TLR3 ligand dsRNA [poly(I:C)] limits CD4 T-cell proliferation, curtailing adaptive immune responses. CD4+ T cells instructed by either lipopolysaccharide (LPS) or poly(I:C)-conditioned DCs promptly upregulated the activation marker CD69. Whereas LPS-pretreated DCs subsequently sustained T-cell clonal expansion, proliferation of CD4+ T cells exposed to poly(I:C)-pretreated DCs was markedly suppressed. This proliferative defect required DC–T cell contact, was independent of IFN-α, and was overcome by exogenous IL-2, indicating T-cell anergy. Coinciding with the downregulation, CD4+ T cells expressed the inhibitory receptor PD-1. Antibodies blocking the PD-1 ligand PD-L1 restored proliferation. dsRNA-stimulated DCs preferentially induced PD-L1, whereas poly(I:C) and LPS both upregulated the costimulatory molecule CD86 to a comparable extent. Poly(dA-dT), a ligand targeting the cytoplasmic RNA helicase pattern-recognition pathway, failed to selectively induce PD-L1 upregulation, assigning this effect to the TLR3 pathway. Poly(I:C)-conditioned DCs promoted accumulation of phosphorylated SHP-2, the intracellular phosphatase mediating PD-1 inhibitory effects. The ability of dsRNA to bias DC differentiation toward providing inhibitory signals to interacting CD4+ T cells may be instrumental in viral immune evasion. Conversely, TLR3 ligands may have therapeutic value in silencing pathogenic immune responses.

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Acknowledgments

This work was funded in part by grants from the National Institutes of Health (RO1 AR 41974, RO1 AR 42527, RO1 AI 57266, RO1 EY 11916, RO1 AG 15043, and UI9 AI 44142), a grant from the Dana Foundation (Dendritic Cell Function in Inflammatory Diseases of Blood Vessels), and a grant from the General Clinical Research Center (MO1 RR00039). The authors thank Tamela Yeargin for manuscript editing.

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The authors declare no competing financial interests.

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Authors and Affiliations

  1. Kathleen B. and Mason I. Lowance Center for Human Immunology, Department of Medicine, Emory University School of Medicine, Room 1003 Woodruff Memorial Research Building, 101 Woodruff Circle, Atlanta, GA, 30322, USA

    Stefan Gröschel, Kisha D. Piggott, Augusto Vaglio, Wei Ma-Krupa, Karnail Singh, Jörg J. Goronzy & Cornelia M. Weyand

  2. Clinical Medicine and Nephrology, University of Parma, Parma, Italy

    Augusto Vaglio

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  1. Stefan Gröschel
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  2. Kisha D. Piggott
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  3. Augusto Vaglio
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Correspondence to Cornelia M. Weyand.

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Stefan Gröschel, Kisha D. Piggott, and Augusto Vaglio have contributed equally to this work.

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Supplemental Fig. 1

TLR4 stimulation cannot revert the TLR3-mediated induction of the inhibitory ligand PD-L1. DC were stimulated with poly(I:C) alone or with a combination of poly(I:C) and LPS. After 4 h, cells were harvested, and PD-L1 transcripts were quantified by real-time PCR. Results are expressed as mean ± SD of triplicate cultures (GIF 20 kb)

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Gröschel, S., Piggott, K.D., Vaglio, A. et al. TLR-mediated induction of negative regulatory ligands on dendritic cells. J Mol Med 86, 443–455 (2008). https://doi.org/10.1007/s00109-008-0310-x

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  • DOI: https://doi.org/10.1007/s00109-008-0310-x

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