interleukin-8 and Cystitis

The innate immune response of urinary tract is critically important in the defense to microbial attack. Toll-like receptor 4 (TLR4) controls initial mucosal response to uropathogenic Escherichia coli (UPEC). However, excessive and dysfunctional TLR signaling may result in severe inflammation and inappropriate tissue damage. Previous studies have demonstrated that single immunoglobulin IL-1R-related receptor/Toll IL-1 receptor 8 (SIGIRR/TIR8) is a member of the toll-interleukin-1 receptor (TIR) family that can negatively modulate TLR4 mediated signaling, but its role in the innate immunity of urinary tract infection remains incompletely defined. In this study, we investigated its cellular distribution and mechanisms involved within the human bladder epithelial cells after LPS stimulation.. Immunostaining, reverse transcription PCR and Western blot results showed that SIGIRR was constitutively expressed in the human bladder epithelial cell lines and was downregulated after LPS stimulation. To further define the role of SIGIRR, cells were transiently transfected with SIGIRR siRNA and stimulated with LPS. SIGIRR gene silencing augmented chemokine expression in response to LPS, as indicated by increased levels of IL-6 and IL-8 secretions in the supernatants compared with negative control siRNA. Furthermore, LPS tolerance, a protective mechanism against second LPS stimulation, was significantly reduced in SIGIRR siRNA transfected cells. Moreover, transient gene silencing augmented LPS-induced NF-κB and MAPK activation.. In conclusion, our results suggest that SIGIRR plays an important role in the negative regulation of LPS response and tolerance in human bladder epithelial cells, possibly through its impact on TLR-mediated signaling.

Topics: Cell Line; Cystitis; Epithelial Cells; Gene Expression; Gene Silencing; Humans; Immune Tolerance; Immunomodulation; Interleukin-6; Interleukin-8; Lipopolysaccharides; Mucous Membrane; Receptors, Interleukin-1; RNA, Small Interfering; Signal Transduction; Toll-Like Receptors; Urinary Bladder

Mast cells have been implicated in bladder inflammation and pathogenesis. To determine if mast cell secretion products can modulate urothelial inflammatory responses we developed an in vitro model of mast cell-urothelial cell interactions.. Cultures of the immortalized urothelial cell line TEU-2 were incubated in the conditioned medium of mast cell cultures. The urothelial inflammatory response to mast cell secretion products was then determined by quantifying nuclear factor kappaB activity, the expression of endogenous nuclear factor kappaB dependent genes and the protein expression of inflammation markers.. Conditioned medium from RBL-2H3 mast cells induced a 4-fold increase in TEU-2 nuclear factor kappaB activity that was independent of the activation state of the mast cells. In contrast, ribonuclease protection assays revealed that the nuclear factor kappaB dependent transcripts tumor necrosis factor-alpha (TNF-alpha), interleukin (IL) 8 and 1beta, and intracellular adhesion molecule 1 (ICAM-1) were induced by mast cell conditioned medium in a manner that strictly depended on mast cell activation (antigen challenge of IgE sensitized RBL-2H3 cells). The dependence on mast cell activation was confirmed by the observation that IL-8 secretion and ICAM-1 protein expression in TEU-2 cultures were induced only by conditioned medium of stimulated RBL-2H3 cells The induction of TEU-2 IL-8 secretion and ICAM-1 expression by mast cell conditioned medium could be blocked by an anti-TNF-alpha antibody or the cysteine protease inhibitor N-acetyl-leucinyl-leucinyl-norleucinal.. Our data support the hypothesis that mast cells may participate in bladder inflammation. Furthermore, TNF-alpha acting via the nuclear factor kappaB signaling pathway may be a mediator of the urothelial response to mast cell secretion products.

Topics: Cell Line; Cystitis; Humans; Intercellular Adhesion Molecule-1; Interleukin-1; Interleukin-8; Mast Cells; NF-kappa B; Signal Transduction; Tumor Necrosis Factor-alpha; Ureter; Urinary Bladder; Urothelium

To estimate canine interleukin-8 (cIL-8) levels in blood plasma samples, a sandwich enzyme linked immunosorbent assay (ELISA) was established. For the development of the sandwich ELISA, polyclonal anti-cIL-8 (capturing), biotinylated anti-cIL-8 (developing) antibodies and glutathione-S-transferase/cIL-8 (GST/cIL-8) fusion protein as an antigen were used. cIL-8 in the fusion protein of GST/cIL-8 was detected in a dose dependent manner. The lowest limit of GST/cIL-8 detectable by this method was 2 ng/ml of GST/cIL-8 (containing; 0.470 ng/ml of cIL-8). IL-8 levels in the plasma samples from apparently healthy dogs were less than 0.470 ng/ ml. Higher levels of IL-8 were detected in the plasma samples of dogs with cystitis, dermatitis, and gastric cancer. These results suggest that the determination of cIL-8 by the sandwich ELISA is useful in diagnosis of inflammatory diseases in dogs.

Topics: Animals; Biomarkers; Cystitis; Dermatitis; Dog Diseases; Dogs; Enzyme-Linked Immunosorbent Assay; Female; Glutathione Transferase; Indicators and Reagents; Inflammation; Interleukin-8; Male; Orchiectomy; Recombinant Fusion Proteins; Reference Values; Renal Insufficiency; Sensitivity and Specificity; Stomach Neoplasms

Topics: Animals; Biomarkers; Cystitis; Female; Humans; Interleukin-8; Rabbits; Urinary Bladder

Neutrophil-activating peptide-1/interleukin-8 (NAP-1/IL-8), secreted by monocytes, macrophages and a number of other cells, acts as a chemoattractant for neutrophil leukocytes and stimulates them to produce a series of responses such as shape change, adherence, exocytosis and respiratory burst, events that are of importance in inflammation. To study the release of NAP-1/IL-8, two human models of inflammation were chosen: transurethral resection of superficial bladder cancer and the subsequent instillation of bacillus Calmette-Guérin (BCG), performed in order to reduce the recurrence rate of papillary bladder tumors. As the secretions of the bladder wall are retained in the urine, patients' urine was collected during 4-hour periods. These urine samples were chromatographed on phosphocellulose. In the elution fractions NAP-1/IL-8 was quantified by a bioassay that measured the elastase release by human neutrophils. The neutrophil-stimulating activity was further purified by reverse phase high performance liquid chromatography. Although no NAP-1/IL-8 activity could be detected in normal individuals, formation of this inflammatory cytokine was observed in patients after transurethral resection and after BCG treatment. The significance and possible use of this secretion are discussed.

Topics: Administration, Intravesical; Aged; Aged, 80 and over; BCG Vaccine; Biological Assay; Chromatography, Liquid; Combined Modality Therapy; Cystitis; Humans; Interleukin-8; Middle Aged; Postoperative Complications; Urethra; Urinary Bladder Neoplasms