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The NLRP3 inflammasome is active but not essential in endotoxin-induced uveitis

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Abstract

Objective

The inflammasome complex involving caspase-1 and nucleotide-binding domain, leucine-rich repeat containing protein (NLRP)3, also known as NALP3 or cryopyrin is important for host responses to microbial pathogens and several autoinflammatory diseases. We investigated the extent to which NLRP3 and caspase-1 control ocular interleukin (IL)-1β production and severity of uveitis (intraocular inflammatory disease) in an established, acute inflammatory uveitis model, endotoxin-induced uveitis (EIU).

Methods

Expression of NLRP3, its adaptor molecule ASC, also known as PYCARD (PYD and CARD domain containing), and caspase-1 were examined by immunoblotting. IL-1β production was measured by enzyme-linked immunosorbent assay (ELISA). Using knockout mice, roles for caspase-1 and NLRP3 were examined in uveitis induced by intraocular injection of Escherichia coli lipopolysaccharide (LPS).

Results

NLRP3, ASC, and caspase-1 proteins are constitutively expressed in eye tissue. During EIU, IL-1β protein production increases; this requires the presence of both caspase-1 and NLRP3. However, severity of EIU is not altered by deficiency in either caspase-1 or NLRP3, as assessed by both intravital microscopy and histology.

Conclusions

These data identify the importance of the NLRP3 inflammasome for IL-1β production in the eye, yet indicate that its participation in EIU is nonessential.

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Acknowledgments

Thanks are due to Dr. Bruce Magun (Oregon Health and Science University) for the transfer of NLRP3 KO mice. This work was made possible by support from NEI/NIH grants (EY019604, EY019020, and EY010572). The authors are grateful for the support from the Stan and Madelle Rosenfeld Family Trust, the William and Mary Bauman Foundation, the Research to Prevent Blindness Foundation, and the American College of Rheumatology Research and Education Foundation.

Author information

Authors and Affiliations

  1. Casey Eye Institute, Oregon Health & Science University, Portland, OR, USA

    Holly L. Rosenzweig, April Woods, Jenna S. Clowers, Stephen R. Planck & James T. Rosenbaum

  2. Department of Molecular Microbiology and Immunology, Oregon Health & Science University, Portland, OR, USA

    Holly L. Rosenzweig

  3. VA Medical Center, Portland, OR, USA

    Jenna S. Clowers

  4. Department of Cell and Developmental Biology, Oregon Health & Science University, Portland, OR, USA

    Stephen R. Planck & James T. Rosenbaum

  5. Department of Medicine, Oregon Health & Science University, Portland, OR, USA

    Stephen R. Planck & James T. Rosenbaum

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  1. Holly L. Rosenzweig
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  2. April Woods
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Correspondence to Holly L. Rosenzweig.

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Responsible Editor: Graham Wallace.

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Rosenzweig, H.L., Woods, A., Clowers, J.S. et al. The NLRP3 inflammasome is active but not essential in endotoxin-induced uveitis. Inflamm. Res. 61, 225–231 (2012). https://doi.org/10.1007/s00011-011-0404-8

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  • DOI: https://doi.org/10.1007/s00011-011-0404-8

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