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Crucial role for the Nalp3 inflammasome in the immunostimulatory properties of aluminium adjuvants

Abstract

Aluminium adjuvants, typically referred to as ‘alum’, are the most commonly used adjuvants in human and animal vaccines worldwide, yet the mechanism underlying the stimulation of the immune system by alum remains unknown. Toll-like receptors are critical in sensing infections and are therefore common targets of various adjuvants used in immunological studies. Although alum is known to induce the production of proinflammatory cytokines in vitro, it has been repeatedly demonstrated that alum does not require intact Toll-like receptor signalling to activate the immune system1,2. Here we show that aluminium adjuvants activate an intracellular innate immune response system called the Nalp3 (also known as cryopyrin, CIAS1 or NLRP3) inflammasome. Production of the pro-inflammatory cytokines interleukin-1β and interleukin-18 by macrophages in response to alum in vitro required intact inflammasome signalling. Furthermore, in vivo, mice deficient in Nalp3, ASC (apoptosis-associated speck-like protein containing a caspase recruitment domain) or caspase-1 failed to mount a significant antibody response to an antigen administered with aluminium adjuvants, whereas the response to complete Freund’s adjuvant remained intact. We identify the Nalp3 inflammasome as a crucial element in the adjuvant effect of aluminium adjuvants; in addition, we show that the innate inflammasome pathway can direct a humoral adaptive immune response. This is likely to affect how we design effective, but safe, adjuvants in the future.

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Figure 1: Aluminium-containing adjuvants stimulate macrophages to produce the pro-inflammatory cytokines IL-1β and IL-18 in a Nalp3 inflammasome-dependent manner.
Figure 2: Caspase-1 activation by aluminium adjuvants requires Nalp3 and ASC.
Figure 3: Alum requires intact endocytic macrophage machinery and causes potassium-gradient-dependent IL-1β secretion without causing significant cell death.
Figure 4: Antibody production and T H 2-dependent inflammation induced by aluminium adjuvants are decreased in the absence of Nalp3, ASC and caspase-1.

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Acknowledgements

We thank L. Zenewicz, Y. Ogura, A. Williams and Y. Wan for discussion and review of this manuscript; A. Coyle, E. Grant and J. Bertin for providing ASC-deficient, Nalp3-deficient and Ipaf-deficient mice; and J. Genzen for providing the P2X7R-deficient mice. This work was supported by the Ellison Foundation, the Bill and Melinda Gates Foundation through the Grand Challenges in Global Health Initiative, and National Institutes of Health grant K08 (F.S.S.). R.A.F. is an Investigator of the Howard Hughes Medical Institute.

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Correspondence to Richard A. Flavell.

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Supplementary Figures

The file contains Supplementary Figures 1-7 with Legends which provide supportive in vitro and in vivo data on the role of Nalp3 in the mechanism of action of aluminum adjuvants. (PDF 157 kb)

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Eisenbarth, S., Colegio, O., O’Connor, W. et al. Crucial role for the Nalp3 inflammasome in the immunostimulatory properties of aluminium adjuvants. Nature 453, 1122–1126 (2008). https://doi.org/10.1038/nature06939

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