This patient experienced an outstanding response to immunotherapy. The patient received several adoptive T cell transfers and regressed from stage IV melanoma to a disease free state and has remained tumor free for several years.

Dendritic cells were matured with TNF alpha, IFN gamma and PGE2. These DC were used to produce a DC vaccine.

We assessed the capacity of the electroporated DCs to activate naive HLA-A2-restricted MelanA-specific CD8(+) T cells without the addition of any exogenous cytokines. A >500-fold increase in MelanA-specific CD8(+) T cells was observed when compared with immature DCs, and a >200-fold increase when compared with cytokine cocktail-matured DCs. In correlation, we found a marked increase in cytolytic and IFN-gamma/tumor necrosis factor-alpha (TNF-alpha) secreting CD8(+) T cells. Our data indicate that immature DCs genetically modified to express stimulating molecul...

A proteomics map of lysosomes purified from non-activated and activated mouse DCs was established.

We found that LPS-stimulated mDCs are able to induce up-regulation of co-stimulatory molecules on co-cultured pDCs. Likewise, CpG-stimulated pDCs activate co-cultured mDCs. The cross talk between these two populations of DC is very efficient since it can be observed at mDC/pDC ratio ranging from 1/10 to 10/1.

We have optimized the MAGE-A3 peptides to load onto HLA-DR*1101 tetramers.

Migration of DCs was studied in vivo in melanoma patients exploring MRI. From isolated lymphnodes obtained after surgery we could identify single DC after staining with Prussian blue for iron.

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.

These DC were matured using CpG-DNA as TLR9 ligand. Mature DCs upregulated CD80, CD86, MHC class II and produced cell type specific cytokines. CD8+ myeloid DCs “cross-presented” exogeneous Ags (Ovalbumin).

Melanoma patients were vaccinated with 4 peptides (MAGE-A3, gp100, NA17 and tyrosinase) presented by HLA-A2 and monitored. We measured responses to these peptides injected with Montanide, not knowing whether the Montanide adjuvant was important for these T cell responses. We therefore injected the same peptides without adjuvant, and no response was observed. We conclude that Montanide had a clear adjuvant effect for the CD8 T cell responses measured against these four peptides.

These are lymph nodes from patients that undergo surgery due to clinical situations that do not affect the immune system (most frequently malformations). We have attempted to identify and characterize CD8T lymphocyte populations in humans. We compared the distribution of phenotypically distinct cell sets using seven color staining in the blood, lymph nodes and spleen. We found that the addition of other markers allow to subdivide N, CM, TEM and TEMRA populations in additional cell types.

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.

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.

CyP was extracted from the freshwater cyanobacterium Oscillatoria Planktothrix FP1. We found that CyP acts as a potent and selective antagonist of bacterial LPS.

Cells are separated into lymphocytes (and monocytes) by elutriation in a closed system (Elutra). [The fraction richest in lymphocytes contained > 90% lymphocytes and viability was > 95%.]

It was found that lymph nodes that drain sites of mature DC or adjuvant inoculation recruited L-selectin- CCR7- effector and memory CD8+ T cells. This cell recruitment required CXCR3 expression on T cells and occurred through high endothelial cells (HEV) in concert with HEV luminal expression of the CXCR3 ligand CXCL9.

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.

The question was addressed of whether bLf, whose immunomodulant properties are now recognized, could influence MDDC differentiation/activation.

We investigated the interactions between human monocyte derived DCs (MDDC), generated in the presence of GM-CSF and IL-4 (IL-4 DC), and antigen-stimulated circulating gamma delta T lymphocytes, bearing the Vgamma2 TCR.

Since LPS-activated DC are potent inducers of NK cell priming and lymph nodes appear to be a key place in which DC and NK cell interactions occur, we have investigated the in vivo capacity of resting and DC-primed NK cells to reach the draining lymph nodes.