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...

We obtained data on the protein changes that occur at the cell surface of a maturing DC, induced by a prototypical maturation stimulus such as LPS. Results obtained from the LC-MS/MS analysis of the integral plasma membrane proteome of D1 cells stimulated for 24 hours with LPS have been compared to the results obtained from immature D1 cells. As expected, DC up-regulated the cell surface expression of all proteins known to be part of the so-called “immunological synapse”, namely MHC class II (region E and K), CD40, CD80, CD86 and ICAM. Interestingly, LPS stimul...

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...

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...