Polarization of primary human monocytes by IFN-gamma induces chromatin changes and recruits RNA Pol II to the TNF-alpha promoter.

Monocyte polarization by IFN-gamma or IL-4 drives a complex series of cellular responses leading to increased intracellular killing (IFN-gamma) or enhanced healing (IL-4) among other functional responses. We studied the effect of IL-4 and IFN-gamma polarization on histone modifications at the TNF-alpha locus in human primary monocytes. IFN-gamma polarization markedly increased the expression of TNF-alpha, whereas IL-4 treatment decreased the expression. We found that IFN-gamma alone increased histone H4 acetylation at the TNF-alpha promoter. The effect of IFN-gamma on TNF-alpha expression was durable upon cytokine washout and even repolarization with IL-4. Concordantly, IFN-gamma-mediated H4 acetylation was also durable. IFN-gamma recruited activating transcription factor-2 via p38 to the TNF-alpha promoter, but inhibition of p38 had minimal effect on H4 acetylation. In a novel finding, we found that IFN-gamma recruited RNA Pol II to the human TNF-alpha promoter via ERK signaling, but did so without initiating transcription, leading to a poised condition. These studies provide an important perspective on monocyte polarization. Polarization by IFN-gamma has a durable effect on TNF-alpha expression, and histone acetylation may provide a mechanism for persistence of the effect.