Modulation of functional characteristics of resident and thioglycollate-elicited peritoneal murine macrophages by a recombinant banana lectin - Fig 7

<p><b>Impact of TLR2- and CD14-triggered mechanisms in production of IL-10 (A, B), TNFα (C, D) and NO (E, F) upon rBanLec stimulation of peritoneal RMs (A, C, E) and TGMs (B, D, F) from BALB/c.</b> RMs and TGMs were stimulated with rBanLec (1, 5 and 10 μg/ml) in the presence of anti-TLR2 or anti-CD14 blocking monoclonal antibodies (20 μg/ml) for 48h. Cytokines and NO were measured in supernatant by ELISA and colorimetric method using Griess reagent, respectively. Bars presented mean concentration ± SE. Corresponding mean concentrations of IL-10, TNFα and NO measured upon incubation under the same conditions but without blocking antibodies are indicated by black solid line and are considered as referent. The significance of the observed differences, due to incubation with particular blocking antibody, was calculated by one-way repeated ANOVA followed by Bonferroni’s multiple comparison test (p <0.05*, p <0.005**, p <0.0001***). LPS–lipopolysaccharide, PEPG–peptidoglycan.</p

Binding of rBanLec to TLR2, TLR4 and CD14 –the effect of methyl-α-D-mannopyranoside.

<p>TLR2, TLR4 and CD14 were extracted from TGMs lysate with specific monoclonal antibodies adsorbed onto microplate. Biotin-labeled rBanLec (10 μg/ml), pre-incubated with or without 0.5 M methyl-α-D-mannopyranoside (α-D-Man), was added to the wells. Alkaline phosphatase / <i>p-</i>nitrophenylphosphate system was used for the visualization of rBanLec binding. Results are presented as mean A₄₀₅ ± SE. The significance of the observed differences was calculated by one-way repeated ANOVA followed by Bonferroni’s multiple comparison test (p <0.05*, p <0.005**, p <0.0001***). Solid lines indicate compared groups.</p