These DC were used to elicit CTL responses in a mouse model.

Indium labelled E/L-S DC (without antigen loading) is administered i.v. to a patient to study the reaction with regard to DC migration.

We are defining the differential roles of intestinal lymph DC subsets in activation of naïve and memory T cells. Two of the three DC subsets are highly potent activators of naïve CD4+ T cells, and do not appear to be constitutively suppressed in any way.

My laboratory continued to analyse the ability of invariant NKT (iNKT) cells to assist priming of antigen specific T and B cell responses. We have demonstrated that activation of human DC by Toll like receptor ligands (TLR-L) modulates the lipid biosynthetic pathway, resulting in enhanced recognition of CD1d-associated lipids by iNKT cells; we have clarified the mechanisms by which CD1d-restricted lymphocytes translate T cell receptor (TCR) recognition of lipids with similar group heads into distinct biological responses; and we demonstrated that pathogen-derive...

Monocytes were isolated from PBMC to generate dendritic cells.

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

It was found that lymph nodes that drain sites of mature DC or adjuvant inoculation recruited L-selectin- CCR7- effector cells.

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.

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.

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%.]

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.

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.

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.

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...

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

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).

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.

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

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

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.