Similarly, a low TLR4 expression or Crenigacestat supplier activity is associated with increased UTI susceptibility. Such strategies would impede pathogen clearance in vivo and cause recurrent UTIs [8, 9]. Lactobacillus is a genus of Gram-positive bacteria naturally found in the healthy human vagina [10] and urethra [11]. Moreover, a low Lactobacillus count is inversely related to high numbers of E. coli in the vagina and a history of recurrent UTI [12]. Several lactobacilli strains are used as probiotics to prevent infections

within the gastrointestinal and urogenital tracts as well as to ameliorate allergic and inflammatory conditions [13–15]. The probiotic mechanisms are believed to include the release of antibacterial substances, biosurfactant production, disruption of biofilms and competitive exclusion [16]. Furthermore, the ability GSK2879552 in vitro of probiotic strains to modulate immunity through NF-κB and mitogen activated protein (MAP) kinase pathways, both important in the development of innate and adaptive immunity, has been reported [17, 18]. Lactobacillus rhamnosus GR-1 is a probiotic isolated from a female urethra [19] used to prevent UTI and bacterial vaginosis, and it has both immunomodulatory and antimicrobial activity [20, 21]. Currently, the immunological effects of lactobacilli on urothelial cells are in large part unexplored. The aim of this current study

was to investigate how L. rhamnosus

GR-1 can affect urothelial immune responses to E. coli. Results Bladder cells responded poorly to lactobacilli compared to heat-killed selleck chemicals E. coli E. coli are potent activators of epithelial immune responses and were therefore used to stimulate activation of NF-κB and cytokine release from bladder cells. After 24 h of challenge with heat-killed E. coli, cells responded with more than 10-fold increase in NF-κB activation compared to resting cells, as measured by the luciferase reporter assay (Figure 1A). Furthermore, challenge gave a substantial increase in pro-inflammatory TNF, IL-6, and CXCL8 levels (Figure 1B, C, and 1D). On the other hand, L. rhamnosus GR-1 was a poor activator of NF-κB. Stimulation with viable lactobacilli led to a Quinapyramine minor increase in the activation of NF-κB while heat-killed bacteria had no significant effect (Figure 2A). Although viable lactobacilli could marginally increase NF-κB activation compared to resting cells, stimulation did not promote release of any of the tested cytokines (TNF, IL-6 and CXCL8). In contrast, it resulted in a small but significant reduction of CXCL8, compared to resting cells, while TNF and IL-6 levels were unaffected (Figure 2B). Figure 1 NF-κB activation and expression of cytokines in bladder cells after E. coli challenge. Bladder cells were stimulated with heat-killed E. coli for 24 h at a concentration corresponding to 108 cfu/ml.