Figure 4.

AM3 treatment does not induce innate immune memory in human monocyte. (A) Experimental design: First 48 hours were conducted as depicted in Figure 1A . Afterwards, adherent cells were left for rest in the base medium supplemented with 10% Human pooled serum for 5 days followed by LPS restimulation. After 24 hours of LPS stimulaion, the supernatants were collected for cytokine measurement. (B) Microscope images at day 6 after the first stimulation, magnification:10x, scale bar: 200 µM. The areas highlighted with a black rectangle show an area of the same image with a larger magnification to better appreciate the cell morphology. (C) Cytokine production after LPS restimulation at day 7 in the innate immune memory model, n=8, Wilcoxon matched-paired test comparing each condition to RPMI. (D) Cytokine production after LPS restimulation at day 7 in the tolerance model. n=18, Friedman test with Dunn’s multiple comparison. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.