The kinetic of the transcriptional profiling of D1 cells treated with either DMSO or LPS was analysed.

PBMC were isolated from buffy coat. Subsequently, monocytes were isolated and differentiated into dendritic cells.

These adherent peripheral blood monocytes are produced from leukapheresis from HLA-A1 and/or HLA-A2 positive patients with stage III/IV melanoma, used to generate DCs from. The latter will subsequently be used to vaccinate the HLA-A1 and/or HLA-A2 positive patients.

These cells are prepared from the removed lesion from patients with stage III or stage IV melanoma and are subsequently used to pulse a portion of the immature dendritic cells.

These cells were used to produce a vaccine for prostate carcinoma and were pulsed with apoptotic allogenic prostate carcinoma cell line LNCap cells.

These dendritic cells were obtained from PBMC from which monocytes had been isolated that differentiated into DC after cell culture.

These DCs have been generated from monocytes after elutriation and have not been pulsed with irradiated tumor cells (apoptotic bodies).

A portion of the DCs is pulsed with irradiated tumor cells (apoptotic bodies) prepared from the removed lesion and matured for 48 hours in presence of TNF-alpha.

Monocytes were cultured in RPMI 1640 medium (GIBCO BRL) supplemented with 2 mM L-glutamine (Sigma), 1% (vol/vol) non-essential amino acids, 100 mM sodium pyruvate, 50 U/ml of penicillin and 50 mg/ml of streptomycin (Gibco BRL) containing 10% (vol/vol) FCS (Hyclone).

These plasmacytoid pDC that were stimulated with both CpG and the TLR-7 ligand R848 were shown to produce very high levels of IL-12p70 (in the ng/ml range per 5 x 105 cells/ml) and IFN-gamma.

These T cells are to be purified using MACS cell separation to contain only CD8+ T cells. The latter are then co-cultured with electroporated DC, stimulated and become evaluated on their cytokine secretion, cytotoxicity and % of antigen specific T cells.

These DC were co-electroporated with constitutively active TLRs and TAA encoding mRNA. They were used to compare different maturation methods.

Human DC exposed to TLR ligands and activated iNKT cells in vitro. They had enhanced expression of maturation markers, suggesting that a cooperative action of TLR ligands and iNKT cells on DC function is a generalizable phenomenon across species. These studies highlight the potential for manipulating the interactions between TLR ligands and iNKT cell activation in the design of effective vaccine adjuvants.

In our effort to demonstrate that pathogen-derived signals to DC mediated via TLRs can be modulated by activated iNKT cells, DC isolated from animals treated simultaneously with TLR and iNKT cell ligands were potent stimulators of naive T cells in vitro compared with these DC from animals treated with the ligands individually.

In our effort to demonstrate that pathogen-derived signals to DC mediated via TLRs can be modulated by activated iNKT cells, these DC isolated from animals treated simultaneously with TLR and iNKT cell ligands were potent stimulators of naive T cells in vitro compared with DC from animals treated with the ligands individually.

Untouched naive CD4+ T cells are co-coltured with fungi -pulsed DCs with the aim to prime naive T cells.

These DC were electroporated, co-cultured with purified CD8 T cells using MACS cell separation to asses the in vitro CD8 T cell stimulatory capacity of DC.

These primed T cells have been obtained from untouched naive CD4+ T cells co-cultured with fungi -pulsed DCs.

Untouched naive CD4+ T cells are co-coltured with fungi -pulsed DCs to prime naive T cells.

Purified CD8 T cells using MACS cell separation were co-cultured with electroporated DC and subequently became evaluated on their cytokine secretion, cytotoxicity and % of antigen specific T cells.