Cytokine production, phenotype and IDO expression by differentially-maturated DC.

<p>A) The DC were generated from DBA/1J mice and cultured for 18 h with LPS, Zymosan or CpG in presence or absence of TE. IL-10 and TNF were detected by ELISA. *<i>P</i><0.05, **<i>P</i><0.005 B) DC were cultured for 18 h with medium, TE, CpG and T/C. Supernatants were tested for IL-12p70, TNF, IL-6, IL-10, IL-23, TGF-β by ELISA. *<i>P</i><0.05, **<i>P</i><0.005 in IL-12p70, TNF, IL-6 and IL-23 versus CpG group, and *<i>P</i><0.05, **<i>P</i><0.005 in IL-10 and TGF-β versus the rest of the groups. C) DC were stimulated as described above and the expression of MHCII and costimulatory molecules (thick lines) was performed by FACS gating the cells on the basis of CD11c⁺ cells. Thin lines show isotype control and values represent the mean fluorescent intensity (MFI). D) Western blot analysis for IDO detection in whole lysates from DC treated as decribed above, using anti-β-actin Ab as the loading control. Data obtained were analyzed by scanning densitometry and were normalized according to the ratio IDO/β-actin expression and these values were compared to medium treated-DC (left panels). L-kynurenine concentration (IDO activity), was determined by a colorimetric assay in lysates or supernatants (right panels) respectively from DC treated as described above. Data shown are representative of two (D) or three (A, B and C) independent experiments. Bars in A, B and D are the means ± SD of triplicate wells per group.