Uroplakin Peptide-Specific Autoimmunity Initiates Interstitial Cystitis/Painful Bladder Syndrome in Mice

Abstract

The pathophysiology of interstitial cystitis/painful bladder syndrome (IC/PBS) is enigmatic. Autoimmunity and impaired urothelium might lead the underlying pathology. A major shortcoming in IC/PBS research has been the lack of an appropriate animal model. In this study, we show that the bladder specific uroplakin 3A-derived immunogenic peptide UPK3A 65–84, which contains the binding motif for IAd MHC class II molecules expressed in BALB/c mice, is capable of inducing experimental autoimmune cystitis in female mice of that strain. A highly antigen-specific recall proliferative response of lymph node cells to UPK3A 65–84 was observed, characterized by selectively activated CD4+ T cells with a proinflammatory Th1-like phenotype, including enhanced production of interferon γ and interleukin-2. T cell infiltration of the bladder and bladder-specific increased gene expression of inflammatory cytokines were observed. Either active immunization with UPK3A 65–84 or adoptive transfer of peptide-activated CD4+ T cells induced all of the predominant IC/PBS phenotypic characteristics, including increased micturition frequency, decreased urine output per micturition, and increased pelvic pain responses to stimulation with von Frey filaments. Our study demonstrates the creation of a more specific experimental autoimmune cystitis model that is the first inducible model for IC/PBS that manifests all of the major symptoms of this debilitating condition.

Chronic pelvic allodynia is mediated by CCL2 through mast cells in an experimental autoimmune cystitis model

“…While numerous animal models have been developed to investigate this disease, our current murine EAC model, generated by bladder-specific UPK3A 65-84, is the first model that manifests both of the major features of IC/PBS, namely, increased urinary frequency (27) and chronic pelvic pain, as assessed by measurement of tactile allodynia. In our previous EAC model, immunization of mice with bladder-specific mouse Ϫ/Ϫ BALB/cJ mice exhibit significantly less tactile allodynia than WT mice after immunization with UPK3A 65-84.…”

“…Knockout mice were bred, and all mice were maintained on a regular 12:12-h light-dark cycle with ad libitum food and water in the Animal Resource Center of Case Western Reserve University. At 8 -10 wk of age, mice were injected subcutaneously with 200 g UPK3A 65-84 in 200 l of an emulsion of equal volumes of water and complete Freund's adjuvant containing 400 g Mycobacteria tuberculosis H37RA (CFA; Difco Laboratories, Detroit, MI) or with an emulsion of water and CFA alone, as previously described (27). Mice were euthanized by asphyxiation with CO 2 followed by cervical dislocation on the number of days after immunization as indicated in the figures.…”

“…In our previously created model of EAC in female BALB/cJ mice, we showed that bladder-specific UPK3A 65-84 peptide induced a CD4-positive T cell-mediated autoimmune response that manifested the major symptoms of human IC/PBS, including increased urinary frequency, decreased urine output per void, and increased referred pelvic pain responses (27). In the present study, we first evaluated the onset and duration of the allodynia induced by UPK3A 65-84 compared with CFAimmunized mice (control) and uninjected naïve mice (baseline).…”

“…We recently created an experimental autoimmune cystitis (EAC) model by injecting a bladder-specific uroplakin 3A-derived immunogenic peptide (UPK3A 65-84) into female BALB/cJ mice (27). The peptide induces CD4-positive T cell-mediated autoimmunity that manifests the urodynamic and pelvic pain phenotypes of human IC/PBS.…”

Retired self-proteins as vaccine targets for primary immunoprevention of adult-onset cancers

“…One important example is HER2 with mRNA expression detected in all of 83 normal human tissue cell types examined, and with protein detectable in many normal human tissues most notably in normal human placenta, breast and bladder [39][40][41]. Considering the substantial levels of HER2 protein expressed in urothelial cells of the normal human bladder (FIGURE 3), it remains unexplained how HER2-targeted immunity can inhibit growth of HER2+ tumors in the absence of any inflammatory consequences associated with autoimmune cystitis including the readily discernible symptoms of increased micturition frequencies and debilitating pelvic pain [94]. This perspective is challenged by those who claim that HER2-targeted adoptive immunotherapy is both effective and safe and occurs in the absence of any observed autoimmunity [95].…”