FIGURE 4.

Cytokine production induced by LAM or hLM in vitro is TLR2-mediated. a, human HEK293 cells transfected with TLR2 were stimulated with 50 μg/ml LAM or hLM. Non-stimulated cells (Medium) or cells stimulated with 50 ng/ml Pam3CSK4 served as a controls. After 24 h at 37 °C, supernatants were harvested and analyzed for IL-8 using ELISA. b, -fold induction of mRNA expression of IL-12p40, IL-12p35, IL-23p19, IL-6, TNF-α, and IL-10 was evaluated by real-time PCR after 6 h of LAM or hLM stimulation (10 μg/ml) of 1 × 10⁶ BL/6 or TLR2^−/− DC, relative to non-stimulated cells. Results are from one representative of two independent experiments. c, production of IL-6, TNF-α, and IL-10 was evaluated by ELISA after 24 h of LAM, hLM (both at 10 μg/ml), or LPS stimulation (25 ng/ml) of 1 × 10⁶ BL/6 and TLR2^−/− DC. Unstimulated DC (Medium) were included as a control. In all cases, results are from one representative of two independent experiments. Each column represents the mean ± S.E. of three wells. Statistical significance was calculated with one-way ANOVA post hoc Tukey's t test (*, p < 0.05; **, p < 0.01; ***, p < 0.001). bld, below detection level.