The CD1d tetramers were generated from in vitro refolded CD1 molecules. The ability to generate CD1d tetramers has provided us with the opportunity of comparing a broad panel of CD1d binding compounds for their ability to stimulate iNKT cells.

4 metastatic melanoma patients were actively vaccinated and additional injections were performed in disease free patients, in patients with stable disease or with slowly progressive disease.

The autologous, apoptotic, leukemic cells (Apo-DC) vaccine was given to 2 cohorts of patients: without additional adjuvants and with GM-CSF as an adjuvant. These two cohorts were monitored for 52 weeks. The clinical and immune monitoring data is being currently analyzed.

The DC are transfected with RNA encoding MelanA, Mage3 and Survivin +/- E/L-selectin. They are used in the DERMA-ER-DC 06 trial.

The moDCs are used in the DERMA-ER-DC 05 trial. They are multipeptide loaded cytokine matured moDC + prior Treg elimination by 3x ONTAK treatment [5µg/kg]. 8 patients are enrolled and receive this substance.

CD1 molecules are refolded in vitro in order to generate CD1 tetramers.

CD1 molecules that are subsequently to be refolded in vitro in order to generate CD1 tetramers.

By immunizing mice with ovalbumin-conjugated anti-Siglec-H Ab in the presence of CpG, we demonstrated generation of antigen-specific CD8 T cells in vivo.

The protein vaccines were injected combined with a combination of iNKT cell-agonist alpha-GalCer and MPL, a detoxified version of LPS that signals through TLR4. This combination treatment stimulates antigen-specific T and B cell responses that are greater than those elicited with alpha-GalCer or MPL alone.

iNKT cells were stimulated in vivo with the synthetic CD1d ligand alpha-GalCer. This significantly enhanced immune responses to protein and peptide based vaccines, due to rapid iNKT dependent DC maturation.

These DC were generated from monocytes after elutriation of leukapheresis product (patients with stage III or stage IV melanoma) and pulsed with irradiated tumor cells (apoptotic bodies) prepared from the removed lesion and matured for 48 hours in presence of TNF-alpha, and subsequently frozen.

These DC had been generated from monocytes after elutriation of leukapheresis product (patients with stage III or stage IV melanoma), were not pulsed and were frozen.

These T cells were obtained from isolation during elutriation and were used for subsequent adoptive transfer.

These T cells were isolated during elutriation of a leukapheresis product (patients with stage III or stage IV melanoma).

DCs were pulsed with allogenic melanoma peptides (SK-Mel 24, HLA-A1 and HLA-A2 positive) plus KLH as immunological tracer and subsequently used to produce a vaccine for melanoma.

TLR ligands (simultaneous activation of TLR4 and TLR8 signaling cascades) were used as stimuli for MDDC and resulted in a marked inhibition of the secretion of the proinflammatory chemokine CCL2 with respect to stimulation through a single TLR.

These dendritic cells were generated from monocytes isolated from buffy coats, activation was induced by lipopolisaccaride (LPS 1microgram/ml, Sigma, St. Louis, MO), by Curdlan (100 micrograms/ml, Wako), by R848 and yeast RNA and by yeast cells in different conditions of culture.

Yeast cells in different conditions of culture (along with R848, Curdland, yeast RNA and LPS) were used to induce DC activation.

Yeast RNA (along with R848, Curdland, LPS and yeast cells in different conditions of culture) was used to induce DC activation.

LPS (1microgram/ml, Sigma, St. Louis, MO) (along with R848, Curdlan, yeast RNA and yeast cells in different conditions of culture) was used to induce DC activation.