Turnover of the extracellular matrix (ECM), activation of macrophages, and accumulation of chemokines/cytokines are all hallmarks of chronic inflammation. Extracellular matrix components, such as hyaluronan (HA), have recently been shown to influence macrophage effector functions, such as the release of inflammatory chemokines and cytokines. Although low mw fragments of the glycosaminoglycan HA induce macrophages to secrete numerous inflammatory mediators, the mechanisms regulating ECM-induced macrophage activation are poorly understood. We have examined the effects of IL-10 and IFN-γ on HA-induced chemokine gene expression in primary mouse macrophages. We found that IL-10 and IFN-γ independently inhibit HA-induced expression of macrophage inflammatory protein-1α (MIP-1α), MIP-1β, and KC at both the mRNA and protein levels. Whereas IL-10 inhibited most of the HA-induced chemokines tested, IFN-γ selectively inhibited only MIP-1α, MIP-1β, and KC. This inhibition did not require prestimulation and occurred even when the cytokines were added up to 3 h after stimulation with HA. For MIP-1α, the inhibition by IFN-γ occurred at the level of transcription, whereas IL-10 predominantly decreased the stability of MIP-1α mRNA. IFN-γ and IL-10 equally inhibited macrophage expression of MIP-1β mRNA at the level of transcription, but MIP-1β mRNA stability was decreased to a greater extent by IL-10. These data identify a previously unrecognized role for IL-10 and IFN-γ as regulators of ECM-induced macrophage expression of inflammatory chemokines.