Figure 2
Effects of CyP priming in experiments of cross-tolerance to E. coli LPS in human primary monocytes. Human primary monocytes isolated by immunomagnetic beads were preincubated with CyP (10 µg/ml) for cross-tolerance induction or E. coli LPS (1 µg/ml) for homogeneous tolerance induction, then washed twice and incubated with E. coli LPS (1 µg/ml) for 5 h. Untreated monocytes (−/−) were maintained in standard medium during the course of the experiment, whereas LPS-activated cells (−/LPS) were maintained in standard medium during the priming phase and then challenged with E. coli LPS (1 µg/ml). Cells incubated twice with CyP (10 µg/ml) were also included in each experimental setting. TNF-α, IL-6, IL-1β, IL-10 were measured in cell culture supernatants by ELISA; mRNA expression was analyzed by RT-PCR and given as fold change over the mRNA level expressed by untreated cells (−/−). The results are shown as mean ± S.E. of at least six independent experiments for cytokines in the supernatants and four independent experiments for mRNA expression *p < 0,05; **p < 0,01; ***p < 0,001 (ANOVA with Tukey’s multiple comparison post hoc test).
