Dataset Information

Human XCR1+ Dendritic Cells Derived In Vitro from CD34+ Progenitors Closely Resemble Blood Dendritic Cells, Including Their Adjuvant Responsiveness, Contrary to Monocyte-Derived Dendritic Cells

ABSTRACT: Functional characterization of human dendritic cell subsets is limited due to the very low frequency of these cells in vivo. We developed an in vitro culture system for the simultaneous generation of XCR1+ DCs and MoDCs from cord blood CD34+ cells. Their global gene expression profiles at steady state and under activation, phenotypes, morphologies and responses to different TLR ligands where characterized and compared with those of their in vivo counterparts. The study demonstrated that the XCR1+ DCs generated in vitro from cord blood CD34+ cells are equivalent to blood XCR1+ DCs and also allowed a rigorous comparison of this DC subset with MoDC which are often considered as the reference model for DCs. Altogether, our results showed that in vitro generated XCR1+ DCs are a better model for the study of blood DC than the conventionally used MoDCs. The different dendritic cell subsets used for the study were either generated in vitro from cord blood CD34+ cells with recombinant human cytokines or isolated from peripheral blood. The subsets were purified by fluorescence-activated cell sorting and their responses to PolyI:C, LPS or R848 were studied including by gene expression profiling.

OTHER RELATED OMICS DATASETS IN: PRJNA247647

ORGANISM(S): Homo Sapiens

SUBMITTER: D Bechlian E Fossum R Chelbi H Liu A Doffin C Caux B Bogen J Valladeau-Guilemond C Sanchez F Montanana-Sanchis T Soos C Asselin-Paturel J Savoret M Shaw Marc Dalod C Chabannon I Perrot M Dalod V Ollion S Balan Tp Vu Manh N Colletti

PROVIDER: E-GEOD-57671 | ArrayExpress | 2014-07-23

SECONDARY ACCESSION(S): GSE57671PRJNA247647

REPOSITORIES: GEO, ArrayExpress

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Human XCR1+ dendritic cells derived in vitro from CD34+ progenitors closely resemble blood dendritic cells, including their adjuvant responsiveness, contrary to monocyte-derived dendritic cells.

Balan Sreekumar S Ollion Vincent V Colletti Nicholas N Chelbi Rabie R Montanana-Sanchis Frédéric F Liu Hong H Vu Manh Thien-Phong TP Sanchez Cindy C Savoret Juliette J Perrot Ivan I Doffin Anne-Claire AC Fossum Even E Bechlian Didier D Chabannon Christian C Bogen Bjarne B Asselin-Paturel Carine C Shaw Michael M Soos Timothy T Caux Christophe C Valladeau-Guilemond Jenny J Dalod Marc M

Journal of immunology (Baltimore, Md. : 1950) 20140709 4

Human monocyte-derived dendritic cell (MoDC) have been used in the clinic with moderately encouraging results. Mouse XCR1(+) DC excel at cross-presentation, can be targeted in vivo to induce protective immunity, and share characteristics with XCR1(+) human DC. Assessment of the immunoactivation potential of XCR1(+) human DC is hindered by their paucity in vivo and by their lack of a well-defined in vitro counterpart. We report in this study a protocol generating both XCR1(+) and XCR1(-) human DC ...[more]

PMID: 25009205

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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.

			Action	DRS
	E-GEOD-57671.additional.1.zip	Other
	E-GEOD-57671.idf.txt	Idf
	E-GEOD-57671.raw.1.zip	Raw
	E-GEOD-57671.raw.2.zip	Raw
	E-GEOD-57671.sdrf.txt	Txt