Immunopathology of interstitial cystitis

This study describes histologic, ultrastructural, and immunofluorescence microscopy findings of cystoscopically directed biopsies of lesional tissue and uninvolved bladder from 308 patients with chronic interstitial cystitis (IC) and no evidence of infection, collagen vascular, or other systemic disease. The majority of cases exhibited strong urothelial cell staining for immunoglobulin A (IgA), which was focal or diffusely involved all layers of the epithelium. Electron microscopy revealed widening of the interstitium between urothelial cells, which may explain leakage of IgA into the urine in cystitis patients. Staining for fibrinogen and IgA was present both in lesional biopsies and uninvolved bladder tissue, suggesting a response to injury affecting the entire bladder urothelium. A minor patient population with idiopathic IC revealed granular IgM (with or without C3) in vessel walls of the lamina propria, suggesting immune complex disease. Ultrastructural evidence of vascular damage included necrosis of endothelial cells, fibrin deposition, and reduplication of basement membranes. Staining for IgE correlated with presence of mast cells and was present in less than 2% of cases, indicating that these cells do not play a prominent role in the pathogenesis of the disease. In the majority of patients with IC mucosal staining for IgA and fibrinogen was a characteristic pattern representing increased IgA secretion and a reaction pattern to injury of unknown cause. A subgroup of patients had evidence of vascular damage and immune complex disease. This study therefore defines two main reaction patterns in interstitial cystitis, which may have implications on prognosis and therapy of the disorder.     

A hypothesis for the etiology of interstitial cystitis based upon inhibition of bladder epithelial repair

Interstitial cystitis (IC) is a chronic bladder disease characterized by distinct bladder mucosal abnormalities, for which the etiology is unknown. Although the epidemiology of this disorder is similar to that of bacterial cystitis, prospective studies using sensitive culture techniques and polymerase chain reaction assay for a variety of microorganisms have failed to identify a specific infectious etiology for IC.We have identified a low-molecular-weight peptide in the urine of IC patients that inhibits the proliferation of normal bladder epithelial cells in vitro. We therefore propose a model of IC, in which this peptide inhibits bladder epithelial regeneration following damage (such as that caused by bacterial cystitis). The chronically damaged epithelium is prone to colonization with various microorganisms, and the resulting exposure to these microorganisms, other urinary antigens, and/or damaged epithelial cells prompts the low-level inflammatory response commonly seen in this disorder.     

Human polyomavirus BK (BKV) load and haemorrhagic cystitis in bone marrow transplantation patients.

Several observations suggest an association between long-lasting haemorrhagic cystitis (HC) in bone marrow transplantation (BMT) recipients and human polyomavirus BK (BKV) reactivation, but no conclusive evidence has been obtained so far. The amount of BKV measured in the urine of BMT patients during an episode of HC was compared with that detected in the urine of BMT patients without HC and of immunocompetent individuals in order to better assess the association of BKV reactivation with HC. For this purpose a quantitative competitive PCR was developed. The application of this assay to clinical samples allowed us to distinguish asymptomatic reactivation both in healthy individuals and in immunocompromised patients from reactivation associated with HC, in almost all cases. Low levels, below the sensitivity of the quantitative assay, were shown in asymptomatic healthy individuals and in about 50% of immunocompromised patients. A significantly higher viral load than in the urine of asymptomatic immunocompromised patients was detected in the urine of patients with HC. These data strengthen the hypothesis that BKV reactivation can cause, together with other factors, the majority of late HC in BMT recipients as well as in patients treated for acute refractory lymphoblastic leukemia.     

Histamine and mucosal mast cells in interstitial cystitis

Interstitial cystitis (IC) is a chronic inflammatory disorder of the urinary bladder of unknown aetiology and pathogenesis. The classic form (Hunner's ulcer) is characterized by high mast cell numbers in the detrusor muscle and an expansion of mucosal mast cells in the lamina propria and also in the epithelium. Such cells can be recovered in bladder washings and in the urine in single cell suspensions. We have counted the mast cells and measured the histamine in bladder washings from 16 patients with classic IC and from a control group of 15 patients with so-called early, non-ulcerative IC. The bladder washings from all patients with classic IC contained well preserved mast cells (median 2.16, range 0.5–8.6×10³ cells/I) and histamine (median 14.3, range 6–66 ng/l), while only occasional mast cells and traces of histamine were found in washings from patients with non-ulcerative IC. The histamine content was strongly correlated to the number of mast cells (r=0.87). The mean histamine content per mast cell was estimated at 7.6±0.65 (SEM) pg/cell. The high histamine content per mast cell in relation to previously published data (2.8–4.6 pg/cell) can be attributed to the mild and rapid handling of the specimens.     

Pharmacological validation of a model of cystitis pain in the mouse

Interstitial cystitis (IC) is a chronic pelvic-perineal pain syndrome of unknown etiology that mainly targets the lower urinary tract. Pain is the most prominent feature of IC and current therapies provide limited relief. Novel treatment options for IC could be identified if more predictive animal models were available. A rat model based on administration of cyclophosphamide (CP) mimics the symptoms of IC and has been well characterized. However, experiments in mice have not consistently reported both the spontaneous and evoked pain behaviors. The current series of studies demonstrate that CP (200-400mg, i.p.) increased both spontaneous and evoked pain behaviors in mice. Additionally, clinically relevant compounds: morphine (1-10mg/kg), ketorolac (1-5.6mg/kg) and duloxetine (3-30mg/kg) all significantly reversed pain behaviors. In contrast, gabapentin (56mg/kg) had no effect. Thus, CP-induced cystitis in mice may be used to evaluate novel therapeutics for the treatment of pain due to interstitial cystitis.     

Serum lipoproteins, antioxidants and urine biochemical constituents in camel cystitis

The present study was undertaken to evaluate the serum and urine biochemical changes in camel cystitis. A total number of 25 male camels (3–12?years old) were subjected to study. Based on the histopathological findings, camels under investigation were classified into three groups: acute cystitis group (N?=?10), chronic cystitis group (N?=?8) and control group (N?=?7). Results revealed significant increases in serum total proteins and globulins in the chronic cystitis group compared with the control and acute cystitis group, significant decrease in serum vitamin C level in the chronic cystitis group compared with the acute cystitis group. In addition, serum β-carotene, α-tocopherol and LDL-C levels were significantly decreased in both the acute and chronic cystitis groups compared with the control group. Furthermore, urine GGT activity significantly increased in the acute cystitis group compared with the control and chronic cystitis groups. In conclusion, the most important biochemical changes in camels with acute and chronic cystitis are decreasing serum α-tocopherol, β-carotene and LDL-C. On the other hand, biochemical findings in chronic cystitis are hyperproteinaemia, hyperglobulinaemia and decrease serum vitamin C level.     

多瘤病毒基因载量与出血性膀胱炎发生相关性的研究

目的研究多瘤病毒的基因载量与异基因造血干细胞移植出血性膀胱炎发生的关系。预防移植后出血性膀胱炎的发生。方法收集40例健康人和40例异基因造血干细胞移植及20例合并出血性膀胱炎患者的血液和尿液，合成多瘤病毒BKV、JCV和SV40基因引物，应用传统的PCR和EvaGreen染料荧光定量PCR法，分别检测血液和尿液多瘤病毒BKV、JCV和SV40DNA。结果40例健康人和40例异基因造血干细胞移植患者血液中BKV、JCV和SV40基因均为阴性；尿液中正常健康人BKV基因的检出率为15％（6／40），JCV基因的检出率为10％（4／40）。异基因造血干细胞移植患者BKV检出率为100％（40／40），JCV基因的检出率为12％（5／40），SV40基因均为阴性。20例出血性膀胱炎患者尿液中BKV基因载量均高于正常人和无出血性膀胱炎的异基因造血干细胞移植患者。结论正常健康人的泌尿系潜伏存在病毒BKV和JCV基因，异基因造血干细胞移植患者出血性膀胱炎的发生与BKV多瘤病毒DNA的载量有关。     

Activation of bladder mast cells in interstitial cystitis.

Interstitial cystitis (IC) is a sterile, inflammatory bladder condition characterized by urinary frequency and urgency, as well as burning and suprapubic pain, which occurs more frequently in women who may suffer for years before diagnosis. An increased number of mast cells have been associated with IC, but the published reports are inconclusive and often conflicting. Human bladder biopsies were analysed blindly for the degree of activation of mast cells in control and IC patients. It was found that mast cells from IC patients averaged as high as 34 cells/mm2 as compared to less than 16/mm2 in controls. Electron microscopy revealed that over 90% of mast cells from IC patients were activated to various degrees. It is concluded that mast cell activation is a pathologic characteristic for IC.     

Serum Peptidome Profiling in Patients with Lung Cancer

Serum peptide profiling is a promising approach for classification of cancer versus noncancer samples. In this study, we aimed to search for discriminating peptide patterns in serum samples between lung cancer patients and healthy controls. The magnetic beads‐based weak cation‐exchange chromatography followed by matrix‐assisted laser desorption and ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) was used in this study to identify patients with lung cancer. In total, serum samples from 64 lung cancer patients (32 for training set and 32 for testing set), 64 healthy controls (32 for training set and 32 for testing set), and 10 COPD patients (for disease control) were analyzed in this study. The mass spectra data analyzed with ClinProTools software was used to distinguish between cancer patients and healthy individuals based on three different algorithm models (GA, SNN, and QC). In the training set, patients with lung cancer could be identified with the mean sensitivity of 98.9% and specificity of100%. Similar results could be obtained from testing set, showing 87% sensitivity and 84.8% specificity. Screening for serum peptide patterns using MALDI‐TOF MS showed high sensitivity and specificity in identifying patients with lung cancer. Anat Rec, 2010. © 2010 Wiley‐Liss, Inc.     

Novel approaches to visualization and data mining reveals diagnostic information in the low amplitude region of serum mass spectra from ovarian cancer patients

The ability to identify patterns of diagnostic signatures in proteomic data generated by high throughput mass spectrometry (MS) based serum analysis has recently generated much excitement and interest from the scientific community. These data sets can be very large, with high-resolution MS instrumentation producing 1-2 million data points per sample. Approaches to analyze mass spectral data using unsupervised and supervised data mining operations would greatly benefit from tools that effectively allow for data reduction without losing important diagnostic information. In the past, investigators have proposed approaches where data reduction is performed by a priori "peak picking" and alignment/warping/smoothing components using rule-based signal-to-noise measurements. Unfortunately, while this type of system has been employed for gene microarray analysis, it is unclear whether it will be effective in the analysis of mass spectral data, which unlike microarray data, is comprised of continuous measurement operations. Moreover, it is unclear where true signal begins and noise ends. Therefore, we have developed an approach to MS data analysis using new types of data visualization and mining operations in which data reduction is accomplished by culling via the intensity of the peaks themselves instead of by location. Applying this new analysis method on a large study set of high resolution mass spectra from healthy and ovarian cancer patients, shows that all of the diagnostic information is contained within the very lowest amplitude regions of the mass spectra. This region can then be selected and studied to identify the exact location and amplitude of the diagnostic biomarkers.     

Cytologic manifestations of cystitis follicularis in urine specimens

Four cases of cystitis follicularis diagnosed by urine cytology are presented, and are the first reported in the cytologic literature. Cystitis follicularis (follicular cystitis) is characterized by formation of lymphoid follicles in the lamina propria of the trigonal region of the bladder, and is considered to be the result of repeated bouts of urinary tract infection, usually bacterial, with other pathologic processes contributing to the development and prolongation of the infection. Cytologically it differs from chronic cystitis with prominent lymphocytosis by the presence of cellular elements from the germinal centers of lymphoid follicles, reminiscent of the cytologic findings in follicular cervicitis, with possible additional epithelial cytologic atypias from the overlying urothelium, which frequently undergoes reactive changes (hyperplastic, metaplastic, and ulcerative). The practical aspect of recognition of this entity in cytologic specimens is avoiding diagnostic errors of possible malignancy (lymphoid or other), and also of other forms of inflammatory disease, such as granulomatous type, with a different clinical significance.     

Neurokinin-1 (NK-1) receptor is required in antigen-induced cystitis

Interstitial cystitis (IC) is a debilitating disease that has been adversely affecting the quality of women's lives for many years. The trigger in IC is not entirely known, and a role for the sensory nerves in its pathogenesis has been suggested. In addition to inflammation, increased mast cell numbers in the detrusor muscle have been reported in a subset of IC patients. Experimentally, several lines of evidence support a central role for substance P and neurokinin-1 (NK-1) receptors in cystitis. The availability of mice genetically deficient in neurokinin-1 receptor (NK-1R(-/-)) allows us to directly evaluate the importance of substance P in cystitis. An unexpected finding of this investigation is that NK-1R(-/-) mice present increased numbers of mast cells in the bladder when compared with wild-type control mice. Despite the increase in mast cell numbers, no concomitant inflammation was observed. In addition, bladder instillation of wild-type mice with a sensitizing antigen induces activation of mast cells and an acute inflammatory response characterized by plasma extravasation, edema, and migration of neutrophils. Antigen-sensitized NK-1R(-/-) mice also exhibit bladder mast cell degranulation in response to antigen challenge. However, NK-1R(-/-) mice are protected from inflammation, failing to present bladder inflammatory cell infiltrate or edema in response to antigen challenge. This work presents the first evidence of participation of NK-1 receptors in cystitis and a mandatory participation of these receptors on the chain of events linking mast cell degranulation and inflammation.     

利用SELDI-TOF质谱技术分析大肠癌患者血清蛋白质谱的变化

目的：研究大肠癌血清蛋白质谱的变化，从而筛选特异性蛋白标志物。 方法： 利用IMAC3蛋白质芯片和SELDI-TOF质谱技术，对64例大肠癌病人和40名正常人的血清蛋白质谱进行分析。获得的蛋白质谱采用Ciphergen公司的Biomarker Wizard和Biomarker Pattern软件分析。 结果： 通过对大肠癌术前血清与正常人血清蛋白质谱分析发现共有19个蛋白质表达量有明显差异。并获得分子量为5 972.67 D、5 927.21 D、6 113.48 D、5 908.55 D和4 292.51 D这5个蛋白质组成的模板，可将大肠癌与正常人正确分组，其正确分组率分别为97.5％（56/64）和80％（32/40）。术后血清蛋白质谱中，原高表达的蛋白质明显下调。 结论： 结果表明通过大肠癌手术前后及正常对照血清中蛋白质谱的比较，筛选得到用以诊断大肠癌的特异性蛋白标志物并用以预后的判断。SELDI-TOF蛋白质芯片技术为建立蛋白质模板从而早期诊断大肠癌提供了可靠的技术平台。     

Corynebacterium group D2 as a cause of alkaline-encrusted cystitis: report of four cases and characterization of the organisms.

In four patients with alkaline-encrusted cystitis, Corynebacterium group D2 was isolated from consecutive urine cultures and stones. Encrusted cystitis occurred in bladders harboring inflammatory or tumorous lesions in patients with chronic or recurrent urinary tract infections appearing after surgery or instrumentation. The urease activity of Corynebacterium group D2 and the neutralization of this enzyme by acetohydroxamic acid are shown. Clinical improvement, disappearance of struvite crystals, and decrease of the urine pH were obtained when these bacteria were eliminated from urine samples. Corynebacterium group D2 strains were highly resistant to many antimicrobial agents but were highly susceptible to norfloxacin and vancomycin when tested at two pHs (7.4 and 8.5).     

RDP58 inhibits T cell-mediated bladder inflammation in an autoimmune cystitis model

Interstitial cystitis (IC) is a chronic inflammatory condition of the urinary bladder with a strong autoimmune component. Currently, the major challenge in IC treatment is the development of effective therapies. RDP58 is a novel d-amino acid decapeptide with potent immunosuppressive activity. In this study, we investigated whether RDP58 was effective as an intravesical agent for treating bladder autoimmune inflammation in a transgenic mouse model (URO-OVA mice). URO-OVA mice were adoptively transferred with syngeneic activated splenocytes of OT-I mice transgenic for the OVA-specific CD8(+) TCR for cystitis induction and treated intravesically with RDP58 at days 0 and 3. Compared with controls, the RDP58-treated bladders showed markedly reduced histopathology and expressions of mRNAs and proteins of TNF-alpha, NGF and substance P. To determine whether the inhibition of bladder inflammation by RDP58 was due to the interference with effector T cells, we treated the cells with RDP58 in vitro. Cells treated with RDP58 showed reduced production of TNF-alpha and IFN-gamma as well as apoptotic death. Collectively, these results indicate that RDP58 is effective for treating T cell-mediated experimental autoimmune cystitis and may serve as a useful intravesical agent for the treatment of autoimmune-associated bladder inflammation such as IC.     

Follicular cystitis in urine cytology: A clinical and cytopathologic study

Despite being an important differential diagnosis of bladder tumor on cystoscopy, follicular cystitis (FC) is rarely diagnosed on cytologic material. We performed a retrospective study on cases of FC diagnosed on bladder biopsy and/or urine cytology in our institution. A total of 35 cases of FC were identified with a female predominance (F:M = 2:1). Hematuria was the most common clinical presentation. Cystoscopic findings included mass lesions, yellow plaques, and surface erythema. History of urinary tract infection was reported in 48% of the patients, and majority of those patients had positive concurrent urine culture, most commonly with beta‐hemolytic streptococcus, Group B. A total of 17 out of 35 patients had urine cytology specimens. When the presence of follicular dendritic cells in clusters of variously sized lymphocytes is used as the cytological diagnostic criterion, 6 out of 17 cases were diagnosed as FC and 5 out of 6 were confirmed by concurrent biopsy. This retrospective study not only analyzed the clinical characteristics of FC but also elucidated the cytological diagnostic criteria of FC and confirmed its specificity.     

Continuous intravesical lidocaine treatment for interstitial cystitis/bladder pain syndrome: safety and efficacy of a new drug delivery device

Limited treatment options exist for patients who suffer from a painful bladder condition known as interstitial cystitis/bladder pain syndrome (IC/BPS). Whether given systemically (orally) or by short-duration (1 to 2 hours) exposure via intravesical instillation, therapeutic agents have exhibited poor efficacy because their concentrations in the bladder are low. A previous attempt to develop a drug delivery device for use in the bladder was unsuccessful, likely as a result of poor tolerability. A continuous lidocaine-releasing intravesical system (LiRIS) was designed to be retained in the bladder and release therapeutic amounts of the drug into urine over a period of 2 weeks. The device was tested in healthy volunteers and IC/BPS patients and was found to be well tolerated in both subject groups because of its small size and freedom of movement within the bladder. The 16 women with IC/BPS who were enrolled in the study met the National Institute of Diabetes and Digestive and Kidney Diseases criteria for bladder hemorrhages or Hunner's lesions. Subjects received either LiRIS 200 mg or LiRIS 650 mg for 2 weeks. Safety, efficacy, cystoscopic appearance of the bladder, and limited pharmacokinetic data were collected. Both doses were well tolerated, and clinically meaningful reductions were seen in pain, urgency, voiding frequency, and disease questionnaires. Cystoscopic examinations showed improvement on day 14 (day of removal) compared with day 1, including resolution of Hunner's lesions in five of six subjects with baseline lesions. Global response assessment showed an overall responder rate of 64% at day 14 and a sustained overall responder rate of 64% 2 weeks later. Extended follow-up suggests that the reduction in pain was maintained for several months after the device was removed.     

YKL‐40 and mast cells are associated with detrusor fibrosis in patients diagnosed with bladder pain syndrome/interstitial cystitis according to the 2008 criteria of the European Society for the Study of Interstitial Cystitis

Richter B, Roslind A, Hesse U, Nordling J, Johansen J S, Horn T & Hansen A B
(2010) Histopathology 57 , 371–383
YKL‐40 and mast cells are associated with detrusor fibrosis in patients diagnosed with bladder pain syndrome/interstitial cystitis according to the 2008 criteria of the European Society for the Study of Interstitial Cystitis Aims: Bladder pain syndrome/interstitial cystitis (BPS/IC), diagnosed according to the new 2008 criteria of the European Society for the Study of Interstitial Cystitis (ESSIC), may lead to detrusor fibrosis. In some inflammatory diseases, fibrosis is related to YKL‐40. The aims were to examine YKL‐40 antigenic expression in bladder tissue and levels in serum and urine in BPS/IC and to evaluate whether YKL‐40 could be a non‐invasive, prognostic biomarker for bladder fibrogenesis and treatment intensity. Methods and results: Immunohistochemistry, immunoelectron microscopy and enzyme‐linked immunosorbent assay (ELISA) analyses in 45 patients showed YKL‐40 expression in detrusor mast cell granules and submucosal macrophages, and elevated YKL‐40 levels in serum and urine compared to healthy individuals (median 72 versus 7 μg/l, P < 0.001). Clinicopathological parameters showed associations of detrusor fibrosis with YKL‐40‐positive cells (P = 0.001), mast cells (P = 0.014) and urine YKL‐40 (P = 0.009). Bladder capacity correlated inversely with YKL‐40‐positive cells (P < 0.001) and mast cells (P = 0.029). Treatment intensity was not associated with YKL‐40. Conclusion: Serum and urine levels of YKL‐40 may be used as non‐invasive biomarkers in BPS/IC for the evaluation of bladder fibrogenesis.     

Boon's disease: Hemorrhagic cystitis in conjunction with massive exfoliation of degenerated urothelial cells (apoptosis?) during intercontinental flights in an otherwise healthy person

Hemorrhagic cystitis is a well-defined clinical emergency, usually occurring in the course of treatment with toxic agents such as cyclophosphamide. We present a case of hemorrhagic cystitis in an otherwise completely healthy female. The three documented attacks were severe and started during intercontinental flights. This type of hemorrhagic cystitis as a disease proved to be a boon, treatable by drinking large amounts of water, and was diagnosed by and in Dr. Boon; thus was the appellation Boon's disease coined. Cellular changes in the urine specimen taken after onset of the disease indicated massive exfoliation of degenerated urothelial cells with morphological features suggestive of apoptosis. It seems likely that this process can be initiated by any event which is associated with compromise of vitality of the urinary bladder lining, such as may occur in hypovolemia. This type of hemorrhagic cystitis is most probably not uncommon in susceptible individuals during intercontinental flights.