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.

MoDC were stimulated with yeast.

An mAb reacting to the immunodominant LACK156-173 peptide of leishmania bound to I-Ad MHC class II molecules was prepared. To this aim, we injected I-Ad/LACK recombinant dimers to TCR transgenic mice which exhibited an increased frequency of LACK-specific T cells. Four out of 600 supernatants, i.e. 2C44, 2F74, 2E60 and 2X8, readily stained LACK156-173-pulsed fibroblasts, but neither unpulsed fibroblasts nor OVA323-339-pulsed cells. BIAcore measurements yielded equilibrium dissociation constants ranging from 1.1 nM for 2C44 mAb to 120 nM for 2F74 mAb.

We have generated a monoclonal antibody, 2C44, that reacted to a peptide derived from the Leishmania LACK protein bound to I-Ad MHC class II molecules. We have successfully used the 2C44 mAb to visualize LACK-presenting DCs in mice infected with the L. major.

These DC were used in an experiment for identification of antigens recognised by T cells from melanoma patients responding to immunotherapy.

Monocyte-derived and cocktail-matured DC electroporated with a combination of RNAs encoding tumor antigens (MelanA, Mage3, Survivin) and E/L-selectin are currently used in a clinical trial.

These DC are exposed to HIV-1 to examine if this can directly modulate their functions or interfere with their cross-talk.

These DC were matured with inflammatory cytokines and subsequently electroporated with mRNA encoding 6 tumor-associated antigens. They were used for DC-based immmunotherapy of melanoma patients.

These monocytes have been obtained from leukapheresis of tissue from patients with stage III or stage IV melanoma (tissue taken from a surgical resection of a melanotic lesion). They will subsequently be used to generate dendritic cells.

These monocytes were obtained after elutriation of a leukapheresis product (patients with stage III or stage IV melanoma) and are used to obtain dendritic cells from.

These monocyte come from an elutriation process from a metastatic lesion from stage III/IV melanoma patients and will be used for the generation of a vaccine and generation of tumour-lysated loaded DC.

Cells from healthy blood donors are separated into monocytes (and lymphocytes) by elutriation in a closed system (Elutra). [The two most monocyte-rich fractions collected contained > 80% monocytes, viability was > 95%.]

Melanoma patients were vaccinated with peptides (MAGE-A3, gp100, NA17 and tyrosinase), with and without montanide as adjuvant. No response was observed when vaccinating without the adjuvant. We conclude that Montanide had a clear adjuvant effect for the CD8 T cell responses measured against these four peptides.

The animal(s) were immunized in vivo with in vitro matured DC and / or treated using a maturation signal.

We have efficiently transfected DC with RNA encoding a functional protein (E/L-Selectin). The mouse E/L-S transfected DC, when given i.v., homed to the lymph nodes, whereas non transfected DC home only to the spleen.

We have studied the immuno-physiology of rat and mouse intestinal dendritic cells. We have used a known adjuvant, E. coli heat-labile toxin (Etx) and a potential adjuvant R-848, a small TLR7/8 ligand.

This is mRNA encoding CD40L, CD70, caTLR4 and one tumorantigen (gp100, Tyrosinase, Mage-C2 or Mage-A3). It was used to electroporate autologous dendritic cells in order to produce a vaccine for advanced melanoma patients.

Stimulation of Mycobacterium tuberculosis (MTb)-antigen specific T cells ex-vivo in the presence of anti-IL-10 antibodies results in their increased proliferation and IFN- production.

These multimers were provided by P. Coulie and used to establish routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry.

These multipeptide loaded cytokine matured moDC +/- CD40L activation were used for the DERMA-ER-DC 04 trial.