In vivo, epithelial cells that line the intestine are intimately associated with lymphocytes, termed intestinal intraepithelial lymphocytes (iIEL). A putative ligand for iIEL on intestinal epithelial cells is CD1d, and recent studies demonstrate a surface form of this molecule exists on intestinal epithelia. At present, it is not known whether CD1d expression is regulated by cytokines in the intestinal microenvironment. Thus we examined the impact of relevant cytokines on CD1d at the level of mRNA and cell surface expression. Using a sensitive whole cell enzyme-linked immunosorbent assay, we assessed the impact of relevant cytokines on CD1d expression on intestinal epithelial cell lines. We were readily able to detect CD1d on the surface of T84 cells, a cryptlike intestinal epithelial cell line. Epithelial cell exposure to human recombinant interferon-gamma (IFN-gamma) resulted in increased CD1d expression in a dose- and time-dependent manner. Polymerase chain reaction amplification of CD1d cDNA revealed a time-dependent induction after exposure to IFN-gamma. This IFN-gamma effect on CD1d expression was cytokine specific and was evident with epithelial cell lines other than T84, including Caco-2 and HT-29 cells. Finally, we were not able to detect significant surface expression of CD1a, CD1b, or CD1c on intestinal epithelial cell lines in the presence or absence of relevant cytokines. These results indicate that CD1d cell surface protein and cellular mRNA, like other major histocompatibility complex-related molecules, is cytokine regulated in intestinal epithelial cell lines.