human epithelial kidney carcinoma a498 cell line  (ATCC)

Journal: Cells

Article Title: Human Renal Fibroblasts, but Not Renal Epithelial Cells, Induce IL-1β Release during a Uropathogenic Escherichia coli Infection In Vitro

doi: 10.3390/cells10123522

Figure Lengend Snippet: IL-1β and LDH release during a UPEC infection. Renal fibroblasts (TK173) and renal epithelial cells (HK-2 and A498) were infected with UPEC strain CFT073 at MOI 1 or MOI 10 for 6 h followed by analysis of IL-1β ( A ) and LDH release ( B ). Data are presented as mean ± SEM ( n = 3 independent experiments). Asterisks over bars denote statistical significance compared to respective unstimulated control (*** p < 0.001).

Article Snippet: Human renal medullary fibroblast cell line TK173 [ ] (kind gift from Professor Anton Jan van Zonneveld, Leiden University, Leiden, Netherlands), human renal epithelial cell line A498 (HTB-44, American Type Culture Collection (ATCC), Manassas, VA, USA) and the human immortalized proximal tubule epithelial cell line HK-2 (ATCC) were used in this study.

Techniques: Infection, Control

Journal: PLoS Pathogens

Article Title: UPEC kidney infection triggers neuro-immune communication leading to modulation of local renal inflammation by splenic IFNγ

doi: 10.1371/journal.ppat.1009553

Figure Lengend Snippet: ( A ) Stimulation of primary DRG cells with LT004 (HlyA+, black circles) and LT005 (HlyA-, black triangles) for 4 h, with capsaicin (unfilled squares) as positive control and uninfected cells (unfilled circles) as negative control. Individual values and means (red bar) are shown, n = 10. * = p<0.05 calculated by one-way ANOVA with Turkey’s correction for multiple comparisons. ( B ) eATP levels in flow-through media from renal epithelial A498 cells infected with CFT073 (HlyA+, black circles), LT002 (HlyA-, black triangles) and ARD371 (LT002 pBAD-HlyA+, grey diamonds), or uninfected (unfilled circles) and uninfected cells exposed to 0.2% arabinose (unfilled diamonds) under flow. Graph shows means ± SD, n = 3 for each condition. # = p<0.05 of CFT073-infected compared to uninfected cells, ## = p<0.05 of CFT073-infected compared to LT002-infected and uninfected cells, and * = p<0.05 of CFT073- and ARD371-infected compared to LT002-infected and uninfected cells, calculated by two-way ANOVA and Turkey´s correction for multiple comparisons. ( C ) Hemolytic activity, measured as arbitrary units (A.U.), in the flow-through media from cells infected with CFT073 (HlyA+, black circles), LT002 (HlyA-, black triangles) or uninfected (unfilled circles). Representative of n = 3, see Figure S2 for repeats. ( D ) Relative Ifng mRNA expression in spleens 4 h after kidney infection with LT005 (black triangles). Data for infection with LT004 (black circles) and sham-infection (PBS-infused, unfilled circles) is re-visualized from for comparison. Expression in relation to the reference gene Gapdh (delta CT) is shown. * = p< 0.05 determined by Kruskal-Wallis and Dunn’s correction. n = 5 in each group.

Article Snippet: The human epithelial cell line A498 (ATCC) was cultured in RPMI-1640 medium (Sigma-Aldrich) with 10% Fetal Bovine Serum and 1% GlutaMAX (ThermoFisher Scientific) or UltraCulture serum free media (Lonza) with 1% GlutaMAX, and incubated at 37°C and 5% CO 2 for 24 h. One day prior to experiments, 2x10 4 -10 5 A498 cells were seeded into a μ-slide VI 0.4 (ibidi).

Techniques: Positive Control, Negative Control, Infection, Activity Assay, Expressing, Comparison

Journal: PLoS Pathogens

Article Title: UPEC kidney infection triggers neuro-immune communication leading to modulation of local renal inflammation by splenic IFNγ

doi: 10.1371/journal.ppat.1009553

Figure Lengend Snippet: ( A-B ) ELISA measurements of ( A ) IL-6 and ( B ) IL-8 release over time from renal epithelial A498 cells under flow, infected with LT004 (black circles), no infection (unfilled circles) or infection with addition of 250 pg/ml IFNγ (grey boxes), n = 3. The IL-8 data from the 5 h timepoint is re-visualized in ( C ) to show statistical significance. ELISA measurement of ( D ) IL-8 and ( E ) IL-6 release over time from A498 cells infected with LT004 under flow (black circles), uninfected (unfilled circles) or infected with the addition of 250 pg/ml IFNγ 1 h post infection (grey boxes), n = 4. Graphs in A-E show means ± SD. Significance was calculated by two-way ANOVA, with Turkey’s correction for multiple comparisons. * = p<0.05 with significance for comparison between uninfected and both LT004 infected and LT004 infected cells co-incubated with 250 pg/ml IFNγ. # = p<0.05 with significance for comparison between uninfected and LT004 infected cells. ( F ) Cxcl1 mRNA expression in sham-splenectomised (circles) or splenectomised (squares) animals, either infected (black symbols) or PBS-infused (unfilled symbols). Gene expression is given in relation to the reference gene Gapdh (delta CT). Individual data points and median (red bar) are shown. n = 3–5 in each group. * = p<0.05 calculated by Kruskal-Wallis analysis and Dunn’s correction.

Article Snippet: The human epithelial cell line A498 (ATCC) was cultured in RPMI-1640 medium (Sigma-Aldrich) with 10% Fetal Bovine Serum and 1% GlutaMAX (ThermoFisher Scientific) or UltraCulture serum free media (Lonza) with 1% GlutaMAX, and incubated at 37°C and 5% CO 2 for 24 h. One day prior to experiments, 2x10 4 -10 5 A498 cells were seeded into a μ-slide VI 0.4 (ibidi).

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Comparison, Incubation, Expressing, Gene Expression

Journal: PLoS ONE

Article Title: Global gene expression profiling and antibiotic susceptibility after repeated exposure to the carbon monoxide-releasing molecule-2 (CORM-2) in multidrug-resistant ESBL-producing uropathogenic Escherichia coli

doi: 10.1371/journal.pone.0178541

Figure Lengend Snippet: A) IL-6 and IL-8 production from A498 renal epithelial cells after stimulation for 6 h with the original ESBL7 isolate or with ESBL7 that have been pre-exposed 20 times to CORM-2 (250 μM) or vehicle (2.5% DMSO). B) Host renal cell cytotoxicity measured as LDH-release during the same conditions as in panel A and normalized to unstimulated and lysed control cells. Data are presented as mean ± SEM from three independent experiments. Asterisk denotes statistical significance (*p<0.05).

Article Snippet: The human renal epithelial cell line A498 (HTB-44) was obtained from American Type Culture Collection (Manassas, USA) and cultured in Dulbecco's modified eagle medium (DMEM, Sigma-Aldrich) containing 10% fetal bovine serum (FBS), 2 mM L-glutamine, 1 mM non-essential amino acids (all from Invitrogen Ltd, Paisley, UK) at 37°C in a 5% CO 2 atmosphere.

Techniques: Control