Autoimmunity to uroplakin II causes cystitis in mice: a novel model of interstitial cystitis

Eur Urol. 2012 Jan;61(1):193-200. doi: 10.1016/j.eururo.2011.06.028. Epub 2011 Jun 29.

Abstract

Background: The pathophysiology of interstitial cystitis (IC) is unknown. Deficits in urothelial cell layers and autoimmune mechanisms may play a role.

Objective: To examine whether immunization of mice with recombinant mouse uroplakin II (rmUPK2), a bladder-specific protein, would provoke an autoimmune response sufficient to create an IC phenotype.

Design, setting, and participants: RmUPK2 complementary DNA was generated, transferred into a bacterial expression vector, and the generated protein was purified. Eight-week-old SWXJ female mice were immunized with rmUPK2 protein via subcutaneous injection of 200μg of rmUPK2 protein in 200μl of an emulsion.

Measurements: Mice were euthanized 5 wk after immunization. Axillary and inguinal lymph node cells were tested for antigen-specific responsiveness and cytokine production, serum isotype antibody titers against rmUPK2 were determined, and gene expression of inflammatory mediators was measured in the bladder and other organs. For functional analysis, mice were placed in urodynamic chambers for 24-h micturition frequency and total voided urine measurements.

Results and limitations: Immunization with rmUPK2 resulted in T-cell infiltration of the bladder urothelium and increased rmUPK2-specific serum antibody responses in the experimental autoimmune cystitis (EAC) mice models compared with controls. The ratio of bladder to body weight was increased in EAC mice. Quantitative reverse transcriptase polymerase chain reaction analysis showed elevated gene expression of tumor necrosis factor α, interferon γ, interleukin (IL)-17A, and IL-1β in bladder urothelium but not in other organs. Evaluation of 24-h micturition habits of EAC mice showed significantly increased urinary frequency (p<0.02) and significantly decreased urine output per void (p<0.021) when compared with control mice.

Conclusions: Our study showed that a bladder-specific autoimmune response sufficient to induce inflammation and EAC occurs in mice following immunization with rmUPK2. EAC mice displayed significant evidence of urinary frequency and decreased urine output per void. Further phenotype characterization of EAC mice should include evidence for pain and/or afferent hypersensitivity, and evidence of urothelial cell layer damage.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Autoimmune Diseases / genetics
  • Autoimmune Diseases / immunology*
  • Autoimmune Diseases / pathology
  • Autoimmune Diseases / physiopathology
  • Autoimmunity*
  • Cells, Cultured
  • Cystitis, Interstitial / genetics
  • Cystitis, Interstitial / immunology*
  • Cystitis, Interstitial / pathology
  • Cystitis, Interstitial / physiopathology
  • Disease Models, Animal
  • Female
  • Gene Expression Regulation
  • Immunohistochemistry
  • Inflammation Mediators / metabolism
  • Injections, Subcutaneous
  • Mice
  • Phenotype
  • RNA, Messenger / metabolism
  • Recombinant Proteins / immunology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Time Factors
  • Urinary Bladder / immunology*
  • Urinary Bladder / pathology
  • Urinary Bladder / physiopathology
  • Urination
  • Urodynamics
  • Uroplakin II / administration & dosage
  • Uroplakin II / immunology*

Substances

  • Inflammation Mediators
  • RNA, Messenger
  • Recombinant Proteins
  • Upk2 protein, mouse
  • Uroplakin II