Membrane incorporation of 22-hydroxycholesterol inhibits chemokine receptor activity

Cell membrane exposure to oxysterols, such as 22-hydroxycholesterol (22-OHC), has previously been shown to induce a suppressive effect on lymphocyte activation. Based on our previous findings that chemokine binding was significantly inhibited by the extraction of membrane cholesterol, we sought to assess the effects of 22-OHC treatment on chemokine ligand-binding and receptor activity. Our results revealed that 22-OHC, but not nonoxidized cholesterol, significantly reduced the binding of both SDF-1alpha and MIP-1beta to human T-cell lines and PBMCs within 1 h of treatment. Incubating the treated cells at 37 degrees C for 1 h reversed a majority of the inhibitory effects on chemokine binding. 22-OHC also inhibited intracellular calcium mobilization and cell migration in response to SDF-1alpha treatment. Interestingly, while the presence of oxysterols in cell membranes significantly inhibits chemokine receptor function, this inhibitory effect does not involve alterations in receptor conformation, expression, or a direct antagonism of chemokine binding. We propose here a novel mechanism for oxysterol-mediated inhibition of chemokine receptor function and the implications for the presence of oxysterols on immune cells.