Abstract
As an approved vaccine adjuvant for use in humans, alum has vast health implications, but, as it is a crystal, questions remain regarding its mechanism. Furthermore, little is known about the target cells, receptors, and signaling pathways engaged by alum. Here we report that, independent of inflammasome and membrane proteins, alum binds dendritic cell (DC) plasma membrane lipids with substantial force. Subsequent lipid sorting activates an abortive phagocytic response that leads to antigen uptake. Such activated DCs, without further association with alum, show high affinity and stable binding with CD4+ T cells via the adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function–associated antigen-1 (LFA-1). We propose that alum triggers DC responses by altering membrane lipid structures. This study therefore suggests an unexpected mechanism for how this crystalline structure interacts with the immune system and how the DC plasma membrane may behave as a general sensor for solid structures.
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Acknowledgements
We thank L. Lanier and C. Lowell (University of California–San Francisco) for providing mouse bone marrows; P. Kubes (University of Calgary) for ICAM-1–, LFA-1– and TLR4-knockout mice; K. Rock (University of Massachusetts Medical School), P. Cresswell (Yale University), G. Dranoff (Harvard Medical School) and R. Yates and J. Deans (University of Calgary) for cell lines, L.J. Shen, M. Ho, Y. Yang and P. Santamaria for manuscript review; C. Olson and T. Fung for statistical software and advice; E. Chau for manuscript editing; F. Oskouie for cell culture assistance; and M. Schoel and W.-D. Xiang for SEM and transmission electron microscopy technical assistance. This work was supported by grants from the US National Institutes of Health to Y.S. and equipment donation from JPK instruments to M.W.A.
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Y.S. designed experiments with input from M.W.A. and wrote the manuscript with assistance from T.L.F. and D.A.M. T.L.F. performed the experiments unless indicated otherwise below. G.N. performed EDS and SEM assays and developed methods for lipid-crystal binding analysis. A.H. and A.D.M. performed CD4-DC binding analysis and EDS and SEM work and contributed to bilayer lipid synthesis. M.D.D. performed antibody induction and cytokine studies with assistance from Y.F. S.M.W., P.Z. and C.C.L. performed aliphatic chain extension on cholesterol and sphingomyelin. M.E.S. and A.V. performed western blotting. E.P. provided Langmuir trough and technical assistance. J.T. and D.A.M. provided inflammasome-deficient mice and technical assistance and consultation. M.W.A. supervised all aspects of work involving AFM.
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Flach, T., Ng, G., Hari, A. et al. Alum interaction with dendritic cell membrane lipids is essential for its adjuvanticity. Nat Med 17, 479–487 (2011). https://doi.org/10.1038/nm.2306
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DOI: https://doi.org/10.1038/nm.2306
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