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 following stimulation with alpha-galactosylceramide (alpha-GalCer)-pulsed dendritic cells (DCs) have been observed. Because little is known about the molecular events on DCs following interaction with alpha-GalCer, we performed gene expression profiling of 2400 genes in monocytes and monocyte-derived immature DCs pulsed with alpha-GalCer (alpha-GalCer-imDCs). Overall, the expression levels of 48 genes were up-regulated and 28 were down-regulated in alpha-GalCer-imDCs. Semiquantitative RT-PCR analysis on monocytes, imDCs, alpha-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 alpha-GalCer, suggesting a novel immunomodulating effect of alpha-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 TNF alpha for two extra days.
Lapteva
N
N
Department of Human Genetics, University of Tokyo, Tokyo, Japan.
Nieda
M
M
Ando
Y
Y
Nicol
A
A
Ide
K
K
Yamaura
A
A
Hatta-Ohashi
Y
Y
Egawa
K
K
Juji
T
T
Tokunaga
K
K
D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000378:metabolism; D003713:Dendritic Cells / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism; D015536:Down-Regulation; D005699:Galactosylceramides / Q000378:metabolism* / Q000502:physiology*; D015870:Gene Expression; D006023:Glycoproteins / Q000378:metabolism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D006801:Humans; D015847:Interleukin-4 / Q000378:metabolism; D009000:Monocytes / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D040121:Semaphorin-3A; D015854:Up-Regulation
D018076:DNA, Complementary; D005699:Galactosylceramides; D006023:Glycoproteins; D012333:RNA, Messenger; D040121:Semaphorin-3A; D015847:Interleukin-4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D003850:Deoxyribonuclease I; D012259:Ribonuclease, Pancreatic