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DNA released from dying host cells mediates aluminum adjuvant activity

Nature Medicine volume 17, pages 996–1002 (2011)Cite this article

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Abstract

Aluminum-based adjuvants (aluminum salts or alum) are widely used in human vaccination, although their mechanisms of action are poorly understood. Here we report that, in mice, alum causes cell death and the subsequent release of host cell DNA, which acts as a potent endogenous immunostimulatory signal mediating alum adjuvant activity. Furthermore, we propose that host DNA signaling differentially regulates IgE and IgG1 production after alum-adjuvanted immunization. We suggest that, on the one hand, host DNA induces primary B cell responses, including IgG1 production, through interferon response factor 3 (Irf3)-independent mechanisms. On the other hand, we suggest that host DNA also stimulates 'canonical' T helper type 2 (TH2) responses, associated with IgE isotype switching and peripheral effector responses, through Irf3-dependent mechanisms. The finding that host DNA released from dying cells acts as a damage-associated molecular pattern that mediates alum adjuvant activity may increase our understanding of the mechanisms of action of current vaccines and help in the design of new adjuvants.

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Figure 1: Alum induces cell death and release of host DNA at sites of injection.
Figure 2: Host DNA released by alum cytotoxicity mediates alum adjuvant activity on humoral and TH2 cell responses.
Figure 3: Alum and host genomic DNA trigger type I IFN secretion and IgE responses through activation of the Tbk1-Irf3 axis.
Figure 4: Irf3 is essential for the boosting of canonical TH2 cells by alum and genomic DNA.
Figure 5: Deficient migration of inflammatory monocytes impairs alum-induced TH2 and IgE responses in Irf3−/− mice.
Figure 6: Alum-induced iMono migration depends on IL-12p40 homodimer signaling.

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Acknowledgements

The authors thank T. Taniguchi (University of Tokyo) and RIKEN BioResource Center for providing Irf3−/− and Irf9−/− mice, and V. Dixit (Genentech) for providing Nlrp3−/− and Casp1−/− mice. We also thank S. Ormenese and R. Stephaan of the Cell Imaging and Flow Cytometry Technological Platform of the Groupe Interdisciplinaire de Génoprotéomique Appliquée for help with fluorescent-activated cell sorting (FACS) analyses; M. Lebrun for help with confocal microscopy; P. Drion and G. Gaudray for animal management; F. Andris, S. Goriely and O. Leo for helpful discussions; and V. Conrath, L. Duwez, R. Fares, C. François, F. Olivier, J. Parisi, F. Perin and I. Sbai for excellent technical and secretarial assistance.

T.M., D.B., C.M. and C.S. are research fellows, and C.J.D. is a postdoctoral fellow of the Fonds National de la Recherche Scientifique (FRS-FNRS; Belgium). This work was supported by grants of the FRS-FNRS, the Belgian Fonds de la Recherche Scientifique Médicale and the Belgian Programme on Interuniversity Attraction Poles (IUAP; FEDIMMUNE, Belgian Science Policy). This work was also partly supported by the Knowledge Cluster Initiative (K.J.I.); a Grant-in-Aid for Scientific Research (K.J.I. and C.C.) of the Japanese Ministry of Education, Culture, Sports, Science and Technology (MEXT); and by the Core Research Evolutionary Science and Technology (CREST) program at the Japan Science and Technology Agency (K.J.I.).

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Author notes

  1. Ken J Ishii, Fabrice Bureau and Christophe J Desmet: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Cellular and Molecular Physiology, Groupe Interdisciplinaire de Génoprotéomique Appliquée, University of Liège, Liège, Belgium

    Thomas Marichal, Denis Bedoret, Claire Mesnil, Catherine Sabatel, Pierre Lekeux, Fabrice Bureau & Christophe J Desmet

  2. Laboratory of Biochemistry, Faculty of Veterinary Medicine, University of Liège, Liège, Belgium

    Thomas Marichal, Denis Bedoret, Claire Mesnil, Catherine Sabatel, Pierre Lekeux, Fabrice Bureau & Christophe J Desmet

  3. World Premier International Immunology Frontier Research Center, Osaka University, Osaka, Japan

    Keiichi Ohata, Kouji Kobiyama, Cevayir Coban, Shizuo Akira & Ken J Ishii

  4. Laboratory of Adjuvant Innovation, National Institute of Biomedical Innovation, Osaka, Japan

    Kouji Kobiyama & Ken J Ishii

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Contributions

T.M., K.J.I., F.B. and C.J.D. designed the experiments; C.C., K.J.I., F.B. and C.J.D. supervised the project; T.M. and D.B. made the initial observation; T.M. did most of the experiments and compiled the data; T.M., K.O. and K.K. did the experiments involving Tbk1/Tnf double-knockout mice, Zbp1−/−, Ifnar2−/− and Mavs−/− mice; C.M. and C.S. did the FACS analyses; S.A. provided the Tbk1/Tnf double-knockout mice and Zbp1−/− mice; P.L., S.A., K.J.I. and F.B. secured funding; K.J.I. and F.B. provided feedback on the manuscript; and C.J.D. wrote the manuscript.

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Correspondence to Ken J Ishii or Christophe J Desmet.

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Marichal, T., Ohata, K., Bedoret, D. et al. DNA released from dying host cells mediates aluminum adjuvant activity. Nat Med 17, 996–1002 (2011). https://doi.org/10.1038/nm.2403

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