Variations in double-positive (IL-6 and TNF-α positive) monocytes and mDCs after dia. <i>E</i>. <i>coli</i> and TLR stimulation on days 14 and 35.

A significant increase in the percentage of monocytes that were simultaneously producing IL-6 and TNF-α was seen after ex vivo stimulation with LPS (200 ng/mL), flagellin (500 ng/mL), or dia. E. coli (1E7 CFU/well) on day 35, comparing to day 14 (A). The average percentage of double-positive monocytes was almost doubled after stimulation with all stimuli in contrast to the RPMI control (B). Following the stimulation with the same stimuli, mDCs showed a significant increase in IL-6 and TNF-α production after exposure to LPS, flagellin, and dia. E. coli (C). Just about all participants showed an increase in double-positive mDCs after dia. E. coli and LPS stimulation, with an average percentage increase of almost 1.5 times (D). For statistical analysis, paired sample t-test was applied. Data are represented in Whisker-plots with a median, 25% and 75% quartile. * p<0.05; ** p<0.01; *** p<0.001,**** p<0.0001. A visual representation of changes in the percentage of IL-6 and TNF-α producing monocytes (E) and mDCs (F) of a representative participant (#10) on day 14 compared to day 35. The percentage of double-positive (top-right quadrant) monocytes increased from 14.3% to 49% (E), whereas double-positive mDCs had an increase from 18.1% on day 14 to 33% on day 35 (F) in this selected participant.