CXCL16 is a chemokine that is expressed in both transmembrane and secreted isoforms. Both variants have been implicated in atherosclerosis. Increased CXCL16 expression on the surface of human aortic smooth muscle cells induced by interferon gamma (IFNγ) signaling results in enhanced oxidized low density lipoprotein uptake and enhanced recruitment of pro-inflammatory cells. Docosahexaenoic acid (DHA), an omega-3 fatty acid, is known to inhibit IFNγ signaling in inflammatory cells. Therefore, we have investigated the effects of DHA treatment on the ability of IFNγ to induce CXCL16 expression in human aortic smooth muscle cells. We observed that DHA treatment significantly reduced IFNγ-induced CXCL16 expression. As a result, the pro-atherosclerotic functions of CXCL16 were also inhibited. Furthermore, IFNγ-induced STAT1 phosphorylation was inhibited by DHA, suggesting a potential mechanism. In conclusion, our data suggest inhibition of IFNγ signaling as one of the mechanisms behind the beneficial effects of DHA during atherosclerosis. These findings may prove to be important in other disease fields that identify IFNγ as a regulator.