Fig. 4

IL-4 reduces CCR7 mRNA synthesis, intracellular and surface protein content in TNF-α-matured DC. a RT-PCR analysis of CCR7 and β-actin mRNA expression in 5 × 10⁴ iDC, IL-4⁻ and IL-4⁺-DC. b Quantification of RT-PCR analysis in (a). Data are expressed as the ratio of expression of CCR7 to β-actin. The results are representative of four independent experiments. c Interleukin-4⁻ and IL-4⁺-DC were fixed, permeabilized and stained with rat PE-anti-CCR7 antibody (bold lines) or its isotype control (thin lines) and analyzed by flow cytometry. The results are representative of six independent experiments. d Mean fluorescence intensity (MFI) of CCR7 intracellular content expressed as the ratio of fluorescence intensity of specific labeling to background staining. The results are the mean ± SEM of the MFI from six independent experiments. e Immature DC were cultured for 0, 8, 24 or 48 h with IL-4 (25 ng/mL) during maturation with GM-CSF and TNF-α. f Immature DC were cultured for 48 h, either in the absence of (0) or with increasing concentrations (0.5, 1, 5, 25 ng/mL) of IL-4 or IL-13, during maturation with GM-CSF and TNF-α. g Immature DC were activated with GM-CSF and TNF-α (20 ng/mL), LPS (50 ng/mL) or sCD40L (250 ng/mL) in the absence or presence of 25 ng IL-4. e–g Cells were stained with PE-anti-CCR7 antibody and analyzed by flow cytometry. The results are the mean ± SEM of the MFI from four to five separate experiments. *P < 0.05, compared to DC activated in the absence of IL-4