Gene expression analysis in human monocytes, monocyte-derived dendritic cells, and alpha-galactosylceramide-pulsed monocyte-derived dendritic cells

In vitro proliferation and functional activation of V alpha 24NKT cells fol lowing stimulation with alpha -galactosylceramide (alpha -GalCer)-pulsed de ndritic cells (DCs) have been observed. Because little is known about the m olecular events on DCs following interaction with alpha -GalCer, we perform ed gene expression profiling of 2400 genes in monocytes and monocyte-derive d immature DCs pulsed with alpha -GalCer (alpha -GalCer-imDCs). Overall, th e expression levels of 48 genes were up-regulated and 28 were down-regulate d in alpha -GalCer-imDCs. Semiquantitative RT-PCR analysis on monocytes, im DCs, alpha -GalCer-imDCs, and mature DCs confirmed the up- and down-regulat ion of the mRNA expression levels of 28 selected genes. Notably, we identif ied the specific up-regulation of mRNA expression levels of ribonuclease A and collapsin response mediator protein upon the stimulation of imDC with a lpha -GalCer, suggesting a novel immunomodulating effect of alpha -GalCer o n imDCs. In this study, we used imDCs prepared by culturing of monocytes wi th GM-CSF and IL-4 for 5 days and mDCs prepared by further culturing of imD Cs with TNF alpha for two extra days. (C) 2001 Elsevier Science.