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Transcription profiling of human monocytes from patients with dominant negative mutations in PPARg and rosiglitazone treated cells to investigate PPARg regulated gene expression in human dendritic cells


ABSTRACT: In order to gain insights into how PPARg regulates different facets of dendritic cell (DC) differentiation, we sought to identify PPARg regulated genes and gene networks in monocyte-derived dendritic cells using global gene expression profiling. We employed an exogenous ligand activation approach using a selective PPARg ligand (rosiglitazone abbreviated as RSG). In addition, we have defined culture conditions in which human serum (HS) induces PPARg activation via a yet uncharacterized endogenous mechanism. We also compared the gene expression profile of developing dendritic cells obtained from patients harboring dominant negative mutations of the PPARg receptor (C114R and C131Y). Experiment Overall Design: Monocytes were cultured for 6, 24 hours or 5 days with 500 U/ml IL-4 and 800 U/ml GM-CSF; cytokine treatment was repeated at day 3. Cells were obtained from 12 healthy individuals (6 biological replicates; the 6 and 24 hours samples were obtained from a single individual but the 5 days samples from a different one). Ligands were added at the beginning of differentiation. The 6 and 24 hours cells were treated with vehicle (DC) or 1 uM rosiglitazone (DC RSG), in the case of 5 day cultured cells 2.5 uM RSG was used. Cells were cultured in RPMI plus 10% FBS, in the case of DC4-DC6 cells were also cultured in human AB serum (DC HS). Finally we also obtained cells from patients harboring point mutations of the PPARg receptor (C114R and C131Y)

ORGANISM(S): Homo sapiens

SUBMITTER: Istvan Szatmari 

PROVIDER: E-GEOD-8658 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

PPARgamma regulates the function of human dendritic cells primarily by altering lipid metabolism.

Szatmari Istvan I   Töröcsik Daniel D   Agostini Maura M   Nagy Tibor T   Gurnell Mark M   Barta Endre E   Chatterjee Krishna K   Nagy Laszlo L  

Blood 20070730 9


Activation of the lipid-regulated nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma) modifies the immunophenotype of monocyte-derived dendritic cells (DCs). However it has not been analyzed in a systematic manner how lipid metabolism and immune regulation are connected at the transcriptional level via this receptor. Here we present the genome-wide expression analyses of PPARgamma-instructed human DCs. Receptor activation was achieved by exogenous, synthetic as well as  ...[more]

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