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The potential repertoire of the innate immune system in the bladder: expression of pattern recognition receptors in the rat bladder and a rat urothelial cell line (MYP3 cells)

  • Urology - Original Paper
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Abstract

Purpose

The urothelium is a frontline sensor of the lower urinary tract, sampling the bladder lumen and stimulating an immune response to infectious and noxious agents. Pattern recognition receptors (PRRs) recognize such agents and coordinate the innate response, often by forming inflammasomes that activate caspase-1 and the release of interleukin-1. We have shown the presence of one PRR (NLRP3) in the urothelia and its central role in the inflammatory response to cyclophosphamide. The purpose of this study was to (1) assess the likely range of the PPR response by assessing the repertoire present in the rat bladder and (2) determine the utility of the MYP3 rat urothelia cell line for in vitro studies by assessing its PPR repertoire and functional responsiveness.

Methods

Immunohistochemistry was performed for seven PPRs (NLRP1, NLRP3, NLRP6, NLRP7, NLRP12, NLRC4 and AIM2) on bladder sections and MYP3 cells. For functionality, MYP3 cells were challenged with the quintessential NLRP3 activator ATP and assessed for caspase-1 activation.

Results

All PPRs examined were expressed in the bladder and localized to the urothelial layer with several also in the detrusor (none in the interstitia). MYP3 cells also expressed all PRRs with a variable intracellular location. ATP-stimulated caspase-1 activity in MYP3 cells in a dose-dependent manner was reduced by knockdown of NLRP3 expression.

Conclusion

The results suggest that the bladder possesses the capacity to initiate an innate immune response to a wide array of uropathological agents and the MYP3 cells will provide an excellent investigational tool for this field.

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Acknowledgments

The authors would like to thank Margret Romano for help with the immunocytochemistry. This work was supported by internal funds from the Medical University of South Carolina.

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Authors and Affiliations

  1. Division of Urology, Department of Surgery, Duke University Medical Center, DUMC Box 3831, Durham, NC, 27710, USA

    Francis M. Hughes Jr. & J. Todd Purves

  2. Department of Urology, Medical University of South Carolina, Charleston, SC, USA

    Francis M. Hughes Jr. & J. Todd Purves

  3. Department of Pathology and Lab Medicine, Medical University of South Carolina, Charleston, SC, USA

    David P. Turner

  4. Department of Pediatrics, Medical University of South Carolina, Charleston, SC, USA

    J. Todd Purves

  5. Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC, USA

    J. Todd Purves

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  1. Francis M. Hughes Jr.
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Correspondence to Francis M. Hughes Jr..

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the Medical University of South Carolina. All experimental procedures were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) at the Medical University of South Carolina and were performed in accordance with the guidelines set forth in the NIH Guide for the Care and Use of Laboratory Animals, published by the Public Health Service of the USA.

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Hughes, F.M., Turner, D.P. & Todd Purves, J. The potential repertoire of the innate immune system in the bladder: expression of pattern recognition receptors in the rat bladder and a rat urothelial cell line (MYP3 cells). Int Urol Nephrol 47, 1953–1964 (2015). https://doi.org/10.1007/s11255-015-1126-6

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