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Liquid-crystalline ordering of antimicrobial peptide–DNA complexes controls TLR9 activation

Nature Materials volume 14, pages 696–700 (2015)Cite this article

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Abstract

Double-stranded DNA (dsDNA) can trigger the production of type I interferon (IFN) in plasmacytoid dendritic cells (pDCs) by binding to endosomal Toll-like receptor-9 (TLR9; refs 1, 2, 3, 4, 5). It is also known that the formation of DNA–antimicrobial peptide complexes can lead to autoimmune diseases via amplification of pDC activation1,2. Here, by combining X-ray scattering, computer simulations, microscopy and measurements of pDC IFN production, we demonstrate that a broad range of antimicrobial peptides and other cationic molecules cause similar effects, and elucidate the criteria for amplification. TLR9 activation depends on both the inter-DNA spacing and the multiplicity of parallel DNA ligands in the self-assembled liquid-crystalline complex. Complexes with a grill-like arrangement of DNA at the optimum spacing can interlock with multiple TLR9 like a zipper, leading to multivalent electrostatic interactions that drastically amplify binding and thereby the immune response. Our results suggest that TLR9 activation and thus TLR9-mediated immune responses can be modulated deterministically.

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Figure 1: Interferon IFN-α production in dendritic cells through TLR9 activation depends on DNA spacing in electrostatic DNA complexes formed with polycations.
Figure 2: Both IFN-inducing and non-IFN-inducing cationic molecules transport DNA to endosomal TLR9.
Figure 3: The presentation of ordered columnar DNA complexes, with specific inter-DNA spacings.
Figure 4: Coarse-grained model of TLR9 binding to DNA bundles exhibits ‘super-selective’ behaviour in agreement with experimental observations.

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Acknowledgements

This work is supported by NSF grants DMR1411329 and DMR1106106, EU grants ARG-ERC-COLSTRUCTION 227758 and ITN-COMPLOIDS 234810, by the Herchel Smith Fund, and by the Slovenian Research Agency through Grant P1-0055, and the Swiss National Science Foundation (FN 310030-144072). X-ray research was conducted at Stanford Synchrotron Radiation Lightsource, SLAC National Laboratory, supported by the US DOE Office of Basic Energy Sciences under Contract No. DE-AC02-76SF00515, the Advanced Light Source, supported by the US DOE Office of Basic Energy Sciences under Contract No. DE-AC02-05CH11231, and at the UCLA CNSI.

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Author notes

  1. Nathan W. Schmidt, Fan Jin, Roberto Lande and Tine Curk: These authors contributed equally to this work.

Authors and Affiliations

  1. Bioengineering Department, Chemistry & Biochemistry Department, California Nano Systems Institute, University of California, Los Angeles, California 90095, USA

    Nathan W. Schmidt, Fan Jin, Wujing Xian, Calvin Lee & Gerard C. L. Wong

  2. Department of Polymer Science and Engineering, Hefei National Laboratory for Physical Sciences at Microscale, CAS Key Laboratory of Soft Matter Chemistry, University of Science and Technology of China, Hefei 230026, China

    Fan Jin

  3. Department of Dermatology, Lausanne University Hospital CHUV, 1009 Lausanne, Switzerland

    Roberto Lande, Loredana Frasca & Michel Gilliet

  4. Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome 00161, Italy

    Roberto Lande & Loredana Frasca

  5. Department of Chemistry, University of Cambridge, Lensfield Road, CB2 1EW Cambridge, UK

    Tine Curk, Daan Frenkel & Jure Dobnikar

  6. Department for Theoretical Physics, Jožef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia

    Jure Dobnikar

  7. International Center for Soft Matter Research, Beijing University of Chemical Technology, Beijing 100029, China

    Jure Dobnikar

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  1. Nathan W. Schmidt
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Contributions

G.C.L.W., M.G., F.J. and N.W.S. conceived the project. R.L. and L.F. performed the cell experiments. F.J., N.W.S., W.X. and C.L. performed SAXS experiments. N.W.S., F.J. and C.L. analysed SAXS data. T.C., J.D. and D.F. conceived the computational model. T.C. performed simulations. G.C.L.W., N.W.S., M.G., D.F., J.D. and T.C. wrote the manuscript. F.J. and R.L. assisted with the methods section.

Corresponding authors

Correspondence to Jure Dobnikar, Michel Gilliet or Gerard C. L. Wong.

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The authors declare no competing financial interests.

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Schmidt, N., Jin, F., Lande, R. et al. Liquid-crystalline ordering of antimicrobial peptide–DNA complexes controls TLR9 activation. Nature Mater 14, 696–700 (2015). https://doi.org/10.1038/nmat4298

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