Figure 4.

HIV neutrophils are characterized by impaired NF-κB activation dependent on the p65-RelA subunit. (A) ICC florescent intensity analysis with double labeling for p65-RelA (green pseudocolor) and IκB (red) in non- and LPS-stimulated neutrophils. The bars represent average fluorescence intensity ± SD calculated from four patients, using at least 100 single cells for each test. (B right panel) Overlap coefficient of p65-RelA vs. IκB and p65-RelA vs. DNA analysis suggests p65-RelA colocalization with its inhibitor IκB which results in disturbance of p65-RelA relocation to the cell nucleus. (B Low panel) ICC 3D projection of nucleus visualizes the lack of NF-κB (p65-RelA) colocalization with DNA after LPS-stimulation in HIV individuals. Green arrows show an overlap of signal from NF-κB (p65-RelA) and DNA in HCs. (C) Total amount of NF-κB binding sites within histone H3K4me3-marked in non-stimulated neutrophils. The bars represent mean ± SD for HCs (n=3) and HIV-infected individuals (n=3). (D) Disturbances in translocation of NF-κB to the nucleus are reflected in the profile of mRNA expression of proinflammatory genes. The mRNA expression of all targets and genes associated with NF-κB are provided in the Supplementary Table S5 .