Our group has produced in-vivo multi-photon microscopy imaging data on the distribution and motility of NK cells by tracking migration of DC-primed NK cells in vivo. Since LPS-activated DC are potent inducers of NK cell priming and lymph nodes appear to be a key place in which DC and NK cell interactions occur, we have investigated the in vivo capacity of resting and DC-primed NK cells to reach the draining lymph nodes. DC –activated NK cells reach the draining lymph node within 24 hours after intravenous injection and persist there for at least 72 hours. NK cell...

Our group has generated pathway data from human DC treated with pathogen derivatives.

We have established an in-depth proteomic map of murine dendritic cell subsets. We have been working in the attempt to overcome the limits in sample availability, exploiting the recent significant improvements of proteomic technology. The results allowed to lower the required sample amount significantly, an essential feature to analyze limited cell populations like dendritic cells. The group completed the measurements of murine dendritic cell subsets and established an in-depth proteomic map of them. These preliminary results show more than 4500 identified proteins...

Data obtained from a proteomic analysis of human exosomes performed by Sebastian AMIGORENA from the DC-THERA consortium allowed us to suspect a number of proteins as a potential candidate target for our hybridoma 13. We have immunized mice with human DC derived-exosomes and generated hybridomas that produce antibodies recognizing at least the human exosomes. Out of 20 hybridomas, only one appears to recognize specifically DC exosomes and not tumor exosomes nor autologous monocytes or DC or PBL. The Ab produced by this N°13 hybridoma is binding to a protein of abo...

Pathway-level data have been obtained from a pilot experiment on yeast cells treated with peptides with potential antifungal action. We have compared results from both genomics and proteomics experiments and search for similarities at the level of pathways and have used Eu.Gene to compare the two different data sources demonstrating the feasibility of using this program to this task. Overall correlation between changes in RNA and protein expression was poor but increased when looking at components of immunologically-relevant pathways and Gene ontology terms. The...

We generated data on the expression by dendritic cells treated with microbial pathogen derivatives.

Agilent and Affymetrix data were obtained from an expression analysis of DC treated +/- poly I:C +/- kinase inhibitors. We have previously shown that activation of Src family kinases is absolutely required for cytokine production in DC stimulated by agonists of several Toll like receptors. Inhibition of these kinases by the specific inhibitors PP1/2 uncoupled cytokine production from the up-regulation of co-stimulatory molecules in DC. We investigated gene expression profiles in DC stimulated with PolyI:C, pretreated or not with PP2, microarray analysis was perfo...

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.

Proteomic data was obtained from a proteomic analysis of DC-derived organelles such as the exosomes. Following the extensive proteomic analysis of immature and mature DC-derived exosomes, ICAM-1 was identified as the major protein on mature DC-derived exosomes, required for their functional activity. In order to functionally validate the proteomic data, the antigen-presenting function has been investigated. Exosomes bearing functional MHC-peptide complexes have been shown to be presented at the surface of recipient DCs as intact "antigen-presenting microdomains", w...

Proteomic map data will be generated for phagosomes formed after phagocytosis through different phagocytic receptors (FcRs, DEC-205, mannose receptor). We will do this by applying a unique methodology dedicated to the identification of phagosome proteins in dendritic cells, under various experimental conditions, that we developed at the Curie Institute. This methodology incorporates a unique sample preparation procedure based on 96 well plate complex sample trypsinization, a nano-LC-based peptide separation coupled to MSMS analysis sequencing. This procedure led us...

Affymetrix data will be gathered through an integrated Functional Genomics approach to study DC maturation with an unprecedented detail. D1 cells will be stimulated with LPS and harvested at different time-points for GeneChip analysis and for tandem mass spectrometry-based analysis of the integral plasma membrane proteome. A protocol for obtaining highly pure integral membrane proteins has already been set up in the last two months in the lab of Paola Castagnoli in collaboration with the lab of Juri Rappsilber at IFOM. A pilot project has already been performed o...

Data has been obtained on expression and regulations of Notch ligands on DC subsets.

Transcriptomic data has been generated on PB-monocytes in healthy controls, RA and SLE.

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.

Data have been obtained on the antigen recognition of DTH infiltrating T cells. To obtain the data, activation of CD4 and CD8 T cells upon recognition of EBV-B cells expressing vaccinal antigens was evaluated by CD137 upregulation, CD107a and CD40L expression using flow cytometry. Luminex technology was used to quantify the TNFalpha and IFNgamma secretion.

Data has been obtained in a study of antigen recognition of blood-derived T cells. In this study, an IFNgamma ELISPOT was done with PBMC loaded with peptide pools, each consisting of 10 15-mers, which overlap with 11 aminoacids.

We can provide data for two experiments composed of 14 microarrays each. To obtain the data, DCs were stimulated with different form of fungi. We perform RNA preparation, labelling, hybridization, and scanning according to the Affymetrix reference protocols and BioPolo (Prof. Ricciardi-Castagnoli and Dott. Francesca Zolezzi) and preliminary data normalization according to AMDA (BioPolo).