Francis M Hughes1,2, David P Turner3, J Todd Purves4,5,6,7. 1. Division of Urology, Department of Surgery, Duke University Medical Center, DUMC Box 3831, Durham, NC, 27710, USA. monty.hughes@duke.edu. 2. Department of Urology, Medical University of South Carolina, Charleston, SC, USA. monty.hughes@duke.edu. 3. Department of Pathology and Lab Medicine, Medical University of South Carolina, Charleston, SC, USA. 4. Division of Urology, Department of Surgery, Duke University Medical Center, DUMC Box 3831, Durham, NC, 27710, USA. 5. Department of Urology, Medical University of South Carolina, Charleston, SC, USA. 6. Department of Pediatrics, Medical University of South Carolina, Charleston, SC, USA. 7. Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC, USA.
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.
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.
Authors: Edward A Miao; Dat P Mao; Natalya Yudkovsky; Richard Bonneau; Cynthia G Lorang; Sarah E Warren; Irina A Leaf; Alan Aderem Journal: Proc Natl Acad Sci U S A Date: 2010-02-01 Impact factor: 11.205
Authors: Jeannette L Tenthorey; Eric M Kofoed; Matthew D Daugherty; Harmit S Malik; Russell E Vance Journal: Mol Cell Date: 2014-03-20 Impact factor: 17.970
Authors: Francis M Hughes; Nathan A Hirshman; Brian M Inouye; Huixia Jin; Eloise W Stanton; Chloe E Yun; Leah G Davis; Jonathan C Routh; J Todd Purves Journal: Diabetes Date: 2018-11-13 Impact factor: 9.461
Authors: Francis M Hughes; Shelby N Harper; Brent D Nosé; Armand Allkanjari; Michael T Zheng; Huixia Jin; J Todd Purves Journal: Am J Physiol Renal Physiol Date: 2021-08-16
Authors: Francis M Hughes; James G Kennis; Melissa N Youssef; Danielle W Lowe; Brooke E Shaner; J Todd Purves Journal: J Clin Cell Immunol Date: 2016-02-29
Authors: Francis M Hughes; Hayden M Hill; Case M Wood; Andrew T Edmondson; Aliya Dumas; Wen-Chi Foo; James M Oelsen; Goran Rac; J Todd Purves Journal: J Urol Date: 2015-12-18 Impact factor: 7.450