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.

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.

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 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.

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%.]

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.

Monocytes were cultured in serum free medium in the presence of IL-4 and GM-CSF, and concentrated cytokines were added.

It was found that lymph nodes that drain sites of mature DC or adjuvant inoculation recruited memory CD8+ T cells.

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.

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

CD83 overexpression on Melan-A/MART-1-specific tumor-infiltrating lymphocytes (TIL) circumvents the need for CD83 expression on DC.

Analysing antigen loading strategies for DC, we found that the priming capacity of MelanA/HLA-A2-specific autologous CD8+ T cells by lipofected DC was higher compared to electroprated DC.

These monocyte-derived DCs received combined stimulation with TLR ligands. They were used in our study on DC activation by pathogen-derived stimuli.

These DC were matured using CpG-DNA as TLR9 ligand. Mature DCs upregulated CD80, CD86, MHC class II and produced cell type specific cytokines. Matured pDCs pDCs produced type 1 Interferons.

Using pharmacological inhibitors, or macrophages and DC from mice carrying a null mutation in MAP kinase signalling molecule(s), we have evidence for a role of certain MAP kinases in the regulation of IL-10 and IL-12, IFN-gamma production.

These macrophages were used to perform a transcriptional profiling. Stimulation was done with different stimuli at different time points in the presence or absence of a kinase inhibitors.

MAGE-A3 specific CD4+ T cells were found also in a high percentage of melanoma patients but CD4+ T cells showed an unpolarized or Th2 skewed phenotype. We have optimized the MAGE-A3 peptides to load onto HLA-DR*1101 tetramers.