Uptake of CNPs promotes a pro-inflammatory phenotype of DCs.
Human and murine DCs (DC2.4) were either left untreated or incubated with 100 μg/ml empty (90/10) or SIINFEKL-loaded 90/10-CNPs for 5, 24 and 48 hours. A) Flow cytometric analysis of CD80, CD86, HLA-DR and PD-L1 cell surface levels in human DCs. Median fluorescence intensity (MFI) ratio was calculated by normalizing MFI of each surface marker to the MFI of its respective isotype control. Then, MFI ratio of CNP treated cells was normalized to untreated cells and expressed as n-fold MFI of untreated cells. B+C) Relative mRNA levels of TGF-β1, IL-10, IL-1β, IL-6 and TNF-α in B) primary human DCs and C) murine DC2.4 cells was determined by RT-qPCR. Expression levels were normalized to expression of the housekeeping gene TBP/GAPDH and normalized to values determined for untreated cells. Data are presented as mean + SEM or median with 75th and 25th percentile of three independent experiments. *p < 0.05.