Collaborative work with P34 (Sica) has been started to study the effects of Japanin on DC exposed to hypoxia (below) and to perform transcriptional profiling of the treated cells. This work, which could not have been undertaken at this stage without support from DC-THERA, has now led to a joint publication in 2008 (Mancino et al.). Transcriptional data are available.

We can provide Illumina microarray data for four experiments composed of 4 microarrays each on DCs stimulated with pathogenic and non pathogenic fungi. RNA preparation, labeling, hybridization on a HT12 array (Illumina), and scanning were performed according to Illumina instructions by Genomics Lab, Wellcome Trust Centre for Human Genetics (Univeristy of Oxford).

Affymetrix and Illumina microarray data is available on 3X106 DCs that were either not stimulated or stimulated with different stimuli. Cells were harvested after 4 hours of stimulation. RNA extraction was done with TRIzol reagent (Invitrogen). microarray hybridization were performed on the basis of technology used. Affymetrix array data files (CEL files) were pre-processed and normalized using the Robust Multichip Average procedure (RMA). Annotations were updated following a procedure devised by Dai and co-workers (Dai et al., 2005). Computation was performed wit...

A large reference ‘golden experiment’ will be carried out in which the transcriptome of DC vaccines of 50-100 patients will be compared and data transcriptomic data will become available.

To understand whether these regulators of gene expression could somehow contribute to the fine tuning of CCL2 expression in MD-DCs, a quantitative microarray approach that detects 723 human mature miR (miRBase 10.1, Dec 2007), was used to measure miR expression profile in these cells, either untreated or stimulated with LPS, R848, or their combination for 8 h. The analysis of the microarray data generated from 4 independent experiments is in progress.

During 2008, we extended all the data regarding TLR3 signaling to TLR8 signaling. Cytokines assays and immunoblotting studies suggest that Src kinases play a crucial role in the control of both MyD88- and TRIF-dependent pathways. We also extended microarray analysis on human MoDC stimulated with R848 (TLR8 agonist), pretreated or not with PP2. In summary, the new generated data on TLR8 triggered gene expression are similar to previous data obtained with TLR3 stimulation, and confirmed that src kinases inhibition is associated to inhibition of key genes in the infl...

Data were obtained from profiling of human monocyte-derived DC exposed to different maturation cocktails and various activators or inhibitors.

Transcriptional data is available on macrophage knockout BALB/c and C57BL/6 mice using different stimuli at different time points in the presence or absence of a kinase inhibitors.

Pathway data is available form a comparative pathways-based analysis of proteomic data performed from same sources.

Data was generated on DC activation by different forms of the non-pathogenic yeast Saccharomyces cerevisiae. A special focus was laid on the response of MDDCs to the non pathogenic fungi.

In our lab, we have gene expression data in T cells available in responses to LCMV-GP33 protein and listeria-Ova. We studied TcR-Tg clones specific of each antigen (P14-GP33 specific, 0T-1-OVA specific), as well as endogenous polyclonal T cells, specific for the same antigens, identified by MHC pentamers loaded with specific peptides. In each individual T cell, we studied the expression of twenty different genes either mediating effector functions, or coding for different receptors involved in T cell differentiation and memory generation. We were surprised to obs...

We have data on the kinetic of the transcriptional profiling of human MoDC treated with IL2 in vitro.

We have generated data on the kinetic of the transcriptional profiling of D1 cells treated with either DMSO or LPS.

Our laboratory has obtained data on the kinetic of the transcriptional profiling of purified mouse DC treated with IL10.

Data has been obtained from an analysis of immature DC and mature DC from two strains of mice: the MyD88 deficient and the wild-type mice.

Our lab obtained data on functional homogeneity from a gene expression analysis at single-cell level for the expression of 8 effector genes from normal donors. Until 2006 we subdivided CD8 PBL from normal donors in 14 different cell sets based in the expression levels and the co-expression of 5 different cell surface markers and studied functional homogeneity by screening cells from each subset at single-cell level for the expression of 8 effector genes (Blood, 2007). Although these studies allowed us isolate some homogeneous cell types, we yet found heterogenei...

We obtained transcriptional and pathway analysis data of human monocyte-derived DCs stimulated with yeast, spheroplasts, pseudohyphae, spores, purified RNA and not purified RNA and LPS. We propose the role of cell wall in the mechanisms of DC-Pathogen interaction as an experiment that could serve as reference in bioinformatics approaches for the reconstruction of DC activation signalling pathways. We have stimulated human Monocyte derived Dendritic Cells (hMoDC) with yeast, spheroplasts, pseudohyphae, spores, purified RNA and not purified RNA, LPS. We have measur...

We can provide one experiment with 32 micorarrays, two experiments with 8 microarrays and one experiment with 12 micorarrays. The yeast oligonucleotide array was constructed using the S. cerevisiae Genome Oligo Set ™ (Operon Technologies, CA, USA) composed of 6240 optimized oligonucleotides (70mers) each representing one yeast gene.

We obtained transcriptomic data by analysing the effect of the inhibition of Src family kinases by the specific inhibitors PP1/2 on the uncoupling cytokine production in DCs stimulated by agonists of several Toll like receptors. Whole transcriptome analysis indicated the transcription factor IRF-8 and 38 other genes including IL-6 as targets of SRC mediated signalling.

Data were obtained from rat DC subsets by transcriptional profiling.