Figure 6: FYT21 inhibits cell activation by various TLR agonists.

NFkB/AP-1 activity of THP1 reporter cells (a) or RAW reporter cells (b) stimulated with the TLR2/4 agonists LPS of P. aeruginosa (Pa) and E. coli (Ec), the TLR2 agonists LTA and zymosan, the TLR5 agonist flagellin, or the TLR9 agonist ODN 1 h before (A>1 h>FYT21) or after (FYT21>1 h>A) incubation with 10 μM FYT21, together with FYT21 after 1 h preincubation (FYT21+A 1 h preincubation) or FYT21 and agonist were mixed in the cell culture well (FYT21+A). (c) THP1 reporter cells were preincubated for 1 h with 10 μM FYT21 followed by stimulation with conditioned medium (CM) of three strains of P. aeruginosa. (d) Cytokine analysis of whole blood stimulated similarly as c with CM of the clinical strain 15159. (e) Effect of FYT21 on stimulation with recombinant TNF-α. Results are means±s.e.m. of 4–9 experiments. (f) Cytokine analysis of blood collected from Balb/c mice (female 10–12 weeks), 4 h after s.c. injection with 100 μl CM of PAOB1 supplemented with 3 units P. aeruginosa elastase or 10 mM Tris buffer. Results are means±s.e.m. of three independent experiments with 3 mice per group. Values are significantly (*P<0.05, **P<0.005 and ***P<0.0005) different from the controls as analysed using a one-way ANOVA with a Dunnett’s multiple comparisons test (a,b), a paired t-test (c–e) or a Mann–Whitney test (f).