Gene Expression Analysis in Human Monocytes, Monocyte-Derived Dendritic Cells, and α-Galactosylceramide-Pulsed Monocyte- Derived Dendritic Cells

Abstract In vitro proliferation and functional activation of Vα24NKT cells following stimulation with α-galactosylceramide (α-GalCer)-pulsed dendritic cells (DCs) have been observed. Because little is known about the molecular events on DCs following interaction with α-GalCer, we performed gene expression profiling of 2400 genes in monocytes and monocyte-derived immature DCs pulsed with α-GalCer (α-GalCer-imDCs). Overall, the expression levels of 48 genes were up-regulated and 28 were down-regulated in α-GalCer-imDCs. Semiquantitative RT-PCR analysis on monocytes, imDCs, α-GalCer-imDCs, and mature DCs confirmed the up- and down-regulation of the mRNA expression levels of 28 selected genes. Notably, we identified the specific up-regulation of mRNA expression levels of ribonuclease A and collapsin response mediator protein upon the stimulation of imDC with α-GalCer, suggesting a novel immunomodulating effect of α-GalCer on imDCs. In this study, we used imDCs prepared by culturing of monocytes with GM-CSF and IL-4 for 5 days and mDCs prepared by further culturing of imDCs with TNFα for two extra days.