Fig. 4

Expression of co-activation markers CD40, CD80 and CD86, proinflammatory cytokines TNFα, IL-12, IL-6 and interferon-β in DCs activated with TLR3 and TLR4 agonists. a-c DCs were transfected with rAd-GFP (100 PFU/cell) and cultivated for 24 h in the presence of 0–10 μg/ml TLR3 (Poly I:C) or TLR4 (LPS, IMM) agonists. Cells were stained with fluorochrom-labeled antibodies specific to CD40 (a), CD86 (b), CD80 (c) and the mean fluorescence of the samples was detected by flow cytometry. d-g DCs were incubated for 2 (d, e) and 7 (f, g) hours in the presence of 0–10 μg/ml agonists of TLR3 (Poly I:C) or TLR4 (LPS, IMM). cDNA was obtained from total RNA extracts of DC and used as a template for quantitative PCR with primers specific for TNF-α (d), IFN-β (e), IL12 (f), and IL6 (g) and β-actin genes. The expression values of cytokine’s genes were normalized with expression of β-actin gene. Values of mRNA expression after activation were normalized to the same values before activation (point 0). Shown are M ± SD, statistically significant (p < 0.05) differences are indicated by asterisks