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Selected Toll-like receptor agonist combinations synergistically trigger a T helper type 1–polarizing program in dendritic cells

Nature Immunology volume 6, pages 769–776 (2005)Cite this article

Abstract

Toll-like receptors (TLRs) sense microbial products and initiate adaptive immune responses by activating dendritic cells (DCs). As pathogens may contain several TLR agonists, we sought to determine whether different TLRs cooperate in DC activation. In human and mouse DCs, TLR3 and TLR4 potently acted in synergy with TLR7, TLR8 and TLR9 in the induction of a selected set of genes. Synergic TLR stimulation increased production of interleukins 12 and 23 and increased the Delta-4/Jagged-1 ratio, leading to DCs with enhanced and sustained T helper type 1–polarizing capacity. Global gene transcriptional analysis showed that TLR synergy 'boosted' only approximately 1% of the transcripts induced by single TLR agonists. These results identify a 'combinatorial code' by which DCs discriminate pathogens and suggest new strategies for promoting T helper type 1 responses.

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Figure 1: Synergistic stimulation of IL-12p70 production by combinations of TLR agonists.
Figure 2: Temporal requirements for TLR synergy.
Figure 3: IFN-γ and CD40L further amplify IL-12p70 production induced by synergistic combinations of TLR agonists.
Figure 4: Synergistic TLR stimulation is required for maximal upregulation of IL-12p35, IL-23p19 and Delta-4.
Figure 5: TLR synergy enhances and sustains the TH1-polarizing capacity of DCs.
Figure 6: Selective effect of TLR synergy on DC gene expression and on IL-1β release.
Figure 7: TLR synergy has an effect on global gene expression only at late time points.
Figure 8: TLR synergy sustains IκBζ expression and c-Jun phosphorylation.

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Acknowledgements

We thank G. Natoli and A. Macagno for critical reading and A. Martín-Fontecha for help with the experiments using mouse DCs. Supported in part by the Swiss National Science Foundation (31-63885), National Institutes of Health (U19AI057266/01) and European Community (Sixth Framework Programme, contract LSHP-CT-2003-503240, Mucosal Vaccines for Poverty-Related Diseases (MUVAPRED)).

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Authors and Affiliations

  1. Institute for Research in Biomedicine, Oncology Institute of Southern Switzerland, Bellinzona, CH-6500, Switzerland

    Giorgio Napolitani, Federica Sallusto & Antonio Lanzavecchia

  2. Laboratory of Experimental Oncology, Oncology Institute of Southern Switzerland, Bellinzona, CH-6500, Switzerland

    Andrea Rinaldi & Francesco Bertoni

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  1. Giorgio Napolitani
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Correspondence to Giorgio Napolitani.

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Supplementary information

Supplementary Fig. 1

Syngergistic TLR stimulation does not affect the extent of MHC and B7 upregulation. (PDF 56 kb)

Supplementary Fig. 2

Exogenous IFN-β has only a modest effect on IL-12p70 production irrespective of the nature of the maturation stimulus. (PDF 26 kb)

Supplementary Fig. 3

The synergistic effect of R848 on LPS induced IL-12 production requires endosomal acidification. (PDF 57 kb)

Supplementary Fig. 4

TLR synergy increases production of TNF, IL-6 and IL-10. (PDF 25 kb)

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Napolitani, G., Rinaldi, A., Bertoni, F. et al. Selected Toll-like receptor agonist combinations synergistically trigger a T helper type 1–polarizing program in dendritic cells. Nat Immunol 6, 769–776 (2005). https://doi.org/10.1038/ni1223

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