Figure 2 PMN induced growth inhibition of ESBL- and non-ESBL-producing E. coli . Growth of MG1655 and CFT073 incubated with PMN (MOI 10) or without PMN (A). Relative growth inhibition of MG1655, CFT073 and the mean relative growth inhibition of susceptible and ESBL-producing E. coli. The relative growth inhibition (delta OD620) is calculated as (absorbance of bacteria-(absorbance of bacteria + PMN)) (B). Data are presented as mean ± SEM (n = 3 independent experiments). Asterisks denote statistical significance (*p < 0.05). Transepithelial migration of PMN evoked

by ESBL- and non-ESBL-producing E. coli A transepithelial migration assay was performed in order to examine PMN migration evoked by the different E. coli strains. The transwell cell monolayer showed low levels of PMN migration in the absence of bacteria (data not shown). NVP-AUY922 All strains evoked PMN migration after 1 h https://www.selleckchem.com/products/ink128.html but there were differences in their ability to attract the PMN (Figure 3A). The ESBL-induced PMN migration was significantly higher 1.6 ± 0.13 fold (p < 0.001) than the migration induced by susceptible strains (Figure 3B). The MG1655 strain induced a significant higher 3.3 ± 0.44 fold (p < 0.001) migration than the CFT073 strain. MG1655 was also shown to attract the largest number of PMN compared to the other strains (Figure 3B). There were no differences observed between ESBL-producing and susceptible strains

in their ability to attract PMN after 3 h (data not shown). Figure 3 PMN migration across a renal epithelial cell line layer in response to ESBL- and non-ESBL-producing E. coli. A498 cells stimulated by the individual bacterial strains (A), and the mean PMN migration across A498 cell layer stimulated with ESBL- and non-ESBL-producing strains, CFT073

and MG1655 (MOI 10) (B). Data are presented as mean ± SEM (n = 3 independent experiments). Asterisks denote statistical significance (***p < 0.001). Epithelial cytokine production evoked by ESBL- and non-ESBL-producing Adenosine E. coli The activation of pro-inflammatory cytokines from urinary tract epithelial cells was evaluated. Both the ESBL-producing and the susceptible strains induced a significant higher IL-6 and IL-8 production from A498 cells compared to unstimulated cells after 6 h. No significant difference was observed between the ESBL- producing and susceptible strains in their ability to induce cytokine production after 3 h (data not shown). The IL-6 and IL-8 production of A498 cells revealed differences between the individual strains (Figures 4A and 5A) and notably, strains that induced high IL-6 production did also induce high IL-8 production. The cytokine production of A498 cells incubated with ESBL-producing strains when grouped together was significantly lower 28 ± 1.9% (IL-6) and 52 ± 3.5% (IL-8) (p < 0.05) compared to cells stimulated with susceptible strains (Figures 4B and 5B).