Antimicrobial peptides such as cathelicidins can modulate inflammation by interfering with TLR function. Small fragment hyaluronan (HA) is released following injury, and is an endogenous ligand for TLR4 as well as CD44. In this study, we examined the interactions of cathelicidin with HA. Cathelicidin inhibited HA induced MIP-2 release from mouse bone marrow derived macrophages in a CD44 dependent manner but did not inhibit MALP2-induced MIP-2 release. This inhibitory activity was more potent than that of a peptide inhibitor of HA binding (Pep-1) and independent of Gi protein coupled or EGF-R signaling, both targets of cathelicidin inhibited HA-induced MIP-2 release. In assay of cell binding to HA, cathelicidins also significantly inhibited this process, suggesting that this antimicrobial peptide can interfere in other membrane binding events mediated by HA. The significance of this inhibition was demonstrated in a skin inflammation model induced by repeated application of 2, 4-dinitrofluorobenzene. This induced an increase in HA at the site of application and was partially CD44 dependent. Camp−/− mice lacking cathelcidin demonstrated a large increase in ear swelling, cell infiltration, and MIP-2 expression compared with wild type mice. These results suggest that cathelicidin has anti-inflammatory activity in skin that may be mediated in part by inhibition of HA-mediated processes.