Figure 2 In vitro effect of different concentrations of PCT on S. typhimurium LPS-induced release of TNFα in human PBMC evaluated by cytokine biochip array. Human PBMC were cultured for 4 h (panel A), and 24
h (panel B) with the following mixtures which had been pre-incubated at 37°C for 30 min : Sterile saline fluid (SF) plus 50 ng/ml PCT (SF + PCT 50); SF plus 500 ng/ml PCT (SF + PCT 500); SF plus 5000 ng/ml PCT (SF + PCT 5000); LPS of S. typhimurium SL1102 (100 ng/ml) plus SF (LPS + SF); LPS (100 ng/ml) plus 50 ng/ml PCT (LPS + PCT 50); LPS (100 ng/ml) plus 500 ng/ml PCT (LPS + PCT 500); LPS (100 ng/ml) plus Combretastatin A4 manufacturer 5000 ng/ml PCT (LPS + PCT 5000). Results are presented as means ± SEM of at least four experiments each carried out in duplicate. Statistical significance between groups was assessed by Student’s t test. A p < 0.05 was considered significant, selleck screening library whereas not significant (n.s.) difference was associated with a p ≥ 0.05. Statistics were performed in comparison with LPS-stimulated PCT-untreated cells (LPS + SF), and the exact significance index is indicated on the top of the horizontal line encompassing the two statistically compared bars. Following 24 hours of incubation, TNFα release stimulated by LPS was significantly diminished when PCT was used at 50 (p = 0.0185), at 500 (p = 0.0240)
and at 5000 ng/ml (p = 0.0253). The levels of MCP-1 were drastically reduced after 4 hours for all the PCT concentrations (Figure 3A). Moreover after 24 hours, the MCP-1 release significantly decreased following both 500 (p = 0.0397) and 5000 ng/ml (p = 0.0116) of PCT (Figure 3B). In the same experimental setting, the LPS-stimulated release of IL-10 showed a dose-dependent MRT67307 datasheet inhibition by PCT at 24 h that was significant at a concentration of 50 (p = 0.0278), 500 (p = 0.0135)
ADP ribosylation factor and 5000 ng/ml (p = 0.0205) of the polypeptide (Figure 4). After 4 hours, this cytokine exhibited slower kinetic. Even though the release of IL-10 by PCT/LPS-incubated PBMC was significantly (p < 0.05) lower than in the supernatant of LPS alone-challenged PBMC, the level of this cytokine was still quite low and perhaps not biologically relevant (data not shown). Figure 3 In vitro effect of different concentrations of PCT on S. typhimurium LPS-induced release of MCP-1 evaluated by cytokine biochip array. Human PBMC were cultured for 4 h (panel A), and 24 h (panel B) with the following mixtures which had been pre-incubated at 37°C for 30 min : Sterile saline fluid (SF) plus 50 ng/ml PCT (SF + PCT 50); SF plus 500 ng/ml PCT (SF + PCT 500); SF plus 5000 ng/ml PCT (SF + PCT 5000); LPS of S. typhimurium SL1102 (100 ng/ml) plus SF (LPS + SF); LPS (100 ng/ml) plus 50 ng/ml PCT (LPS + PCT 50); LPS (100 ng/ml) plus 500 ng/ml PCT (LPS + PCT 500); LPS (100 ng/ml) plus 5000 ng/ml PCT (LPS + PCT 5000). Results are presented as means ± SEM of at least four experiments each carried out in duplicate.