Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptome analysis of primary human monocyte differentiation

ABSTRACT: In order investigate the control of genes encoding cytoskeletal motor proteins and their interaction partners in the activation program of primary monocytes, we performed whole transcriptome microarray profiling of ex vivo monocyte differentiation to activated macrophages or dendritic cells in monocytes isolated from three anonymous blood donors. Monocytes were isolated from the buffy coats of three anonymous blood donors and differentiated into activated macrophages or activated dendritic cells (DCs) ex vivo. Total RNA from monocytes, activated macrophages or DCs was isolated and submitted for whole transcriptome microarray analysis.

ORGANISM(S): Homo sapiens

SUBMITTER: Zoltan Maliga

PROVIDER: E-GEOD-33273 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Dendritic cells (DCs) are the sentinels of the mammalian immune system and they undergo a complex maturation process mediated by activation upon pathogen detection. Recent studies described the analysis of activated DCs by transcriptional profiling, but translation regulation was never taken in account. Therefore, the nature of the mRNAs being translated at various stages of DC activation was determined with the help of translational profiling, which is the sucrose gradient fractionation of polysomal-bound mRNAs combined to microarrays analysis. Total and polysomal-bound mRNA populations were compared in immature (0h) and LPS-stimulated (4h and 16h) human monocyte-derived DCs with the help of Affymetrix microarrays. Biostatistical analysis indicated that 296 mRNA molecules are translationally regulated during DC-activation. The most abundant biological process among the regulated mRNAs was protein biosynthesis, indicating the existence of a negative feedback loop regulating translation. Interestingly, a cluster of 17 ribosomal proteins were part of the regulated mRNAs, indicating that translation may be fine-tuned by particular components of the translational machinery. Our observations highlight the importance of translation regulation during the immune response, and may favour the identification of novel gene clusters or protein networks relevant for immunity. Our study also provides information on the possible absence of correlation between gene expression and real protein production in DCs. To identify translationally regulated mRNA molecules, gene expression derived from the polysome-bound mRNAs was compared by Affymetrix microarrays analysis to the gene expression derived from unfractionated total mRNAs derived from whole-cell lysates, as recently described on several reports (Johannes 1999, Rajasekhar 2003, Bushell 2006, Lü 2006, Parent 2008). Polysomal RNA (P) and total RNA (T) were isolated from MoDCs generated from four different blood donors. Since three timepoints (0h, 4h and 16h) were chosen for each blood donor and RNA type, twenty-four RNA samples were totally analyzed by microarrays.

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Transcriptome analysis of primary human monocyte differentiation

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