NLRP3/Cryopyrin Is Necessary for Interleukin-1β (IL-1β) Release in Response to Hyaluronan, an Endogenous Trigger of Inflammation in Response to Injury*
- Kenshi Yamasaki‡,
- Jun Muto‡,
- Kristen R. Taylor‡1,
- Anna L. Cogen‡,
- David Audish‡,
- John Bertin§2,
- Ethan P. Grant§3,
- Anthony J. Coyle§4,
- Amirhossein Misaghi¶,
- Hal M. Hoffman¶∥ and
- Richard L. Gallo‡5
- ‡Division of Dermatology, University of California, and Veterans Affairs San Diego Health Care System, San Diego, California 92161, the ¶Department of Pediatrics, University of California, San Diego, California 92161, the ∥Ludwig Institute for Cancer Research, San Diego, California 92161, and §Millennium Pharmaceuticals, Cambridge, Massachusetts 02139
- 5 To whom correspondence should be addressed: MC 9111B, 3350 La Jolla Village Dr., San Diego, CA 92161. Tel.: 858-642-3504; Fax: 858-642-1435; E-mail: rgallo{at}ucsd.edu.
Abstract
Inflammation under sterile conditions is a key event in autoimmunity and following trauma. Hyaluronan, a glycosaminoglycan released from the extracellular matrix after injury, acts as an endogenous signal of trauma and can trigger chemokine release in injured tissue. Here, we investigated whether NLRP3/cryopyrin, a component of the inflammasome, participates in the inflammatory response to injury or the cytokine response to hyaluronan. Mice with a targeted deletion in cryopyrin showed a normal increase in Cxcl2 in response to sterile injuries but had decreased inflammation and release of interleukin-1β (IL-1β). Similarly, the addition of hyaluronan to macrophages derived from cryopyrin-deficient mice increased release of Cxcl2 but did not increase IL-1β release. To define the mechanism of hyaluronan-mediated activation of cryopyrin, elements of the hyaluronan recognition process were studied in detail. IL-1β release was inhibited in peritoneal macrophages derived from CD44-deficient mice, in an MH-S macrophage cell line treated with antibodies to CD44, or by inhibitors of lysosome function. The requirement for CD44 binding and hyaluronan internalization could be bypassed by intracellular administration of hyaluronan oligosaccharides (10–18-mer) in lipopolysaccharide-primed macrophages. Therefore, the action of CD44 and subsequent hyaluronan catabolism trigger the intracellular cryopyrin → IL-1β pathway. These findings support the hypothesis that hyaluronan works through IL-1β and the cryopyrin system to signal sterile inflammation.
Footnotes
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↵6 The abbreviations used are: HA, hyaluronan; fl-HA, fluorescein-labeled hyaluronan; LPS, lipopolysaccharide; TLR, Toll-like receptor; NLR, nucleotide-binding domain and leucine rich repeat-containing; IL-1β, interleukin 1 β; MIP-2, macrophage inflammatory protein-2; FCAS, familial cold autoinflammatory syndrome; PAMP, pathogen-associated molecule pattern; PBS, phosphate-buffered saline; Tricine, N-[2-hydroxy-1,1-bis(hydroxymethyl) ethyl]glycine; siRNA, small interference RNA; DMSO, dimethyl sulfoxide; FCS, fetal calf serum; ELISA, enzyme-linked immunosorbent assay; WT, wild type; RT-PCR, reverse transcription-PCR.
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↵* This work was supported, in whole or in part, by National Institutes of Health Grant P01-HL057345 (to R. L. G.).
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The on-line version of this article (available at http://www.jbc.org) contains supplemental methods and two supplemental figures.
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↵1 Present address: The Scripps Research Institute Dept. of Immunology, IMM-8 10550 North Torrey Pines Rd., La Jolla, CA 92037.
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↵2 Present address: GlaxoSmithKline, 1250 South Collegeville Rd., Collegeville, PA 19426.
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↵3 Present address: Schering-Plough, 2000 Galloping Hill Rd., Kenilworth, NJ 07033.
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↵4 Present address: MedImmunie, LLC, One MedImmune Way, Gaithersburg, MD 20878.
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- Received August 3, 2008.
- Revision received February 18, 2009.
- The American Society for Biochemistry and Molecular Biology, Inc.
