Fig. 6.

Intracellular cytokine responses of DC populations to TLR ligation. MLN from C57BL/6 mice were taken 7 days following infection with N. brasiliensis (Nb) or H. polygyrus (Hp). Organs were harvested, treated with liberase CI for 25 min and EDTA for 5 min, then homogenised and labelled with CD11c microbeads (Miltenyi) before selecting on a LS column. Following selection, CD11c⁺ cells were counted, plated at 10⁵/well with 1 μg/ml LPS and 20 μg/ml brefeldin A for 6 h. Cells were then harvested and stained intracellularly with PE-conjugated IL-6, TNF-α, IL-12p40/70, or an isotype control (IgG1). Left hand panels show representative flow cytometry plots of intracellular staining against autofluorescence measured in the FITC channel, for LPS-stimulated DCs; right hand panels show data from individual mice, and include unstimulated control cells cultured in the absence of LPS. ^*p < 0.05, ^**p < 0.01, ^***p < 0.001. Data presented are representative of two experiments with similar results.