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...

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.

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.

Laminarin (500 micrograms/ml) was added to DCs in order to block their beta-glucan receptors.

There are lentiviral vectors available in the lab.

These mice were used for transcriptional profiling of macrophages stimulated with different stimuli at different time points in the presence or absence of a kinase inhibitors.

These mice were used for transcriptional profiling of macrophages stimulated with different stimuli at different time points in the presence or absence of a kinase inhibitors.

The leukapheresis product is taken from a HLA-A2 positive healthy donor and is subsequently separated into monocytes and lymphocytes.

In a study aiming at the identification of molecular signatures for alternatively activated DC it was found that AA-DC induced in the presence of LPS and IL-10 also produced high levels of the long pentraxin PTX3. DC produced conspicuous amounts of PTX3 when stimulated with LPS and this production is further increased in the presence of IL-10. However, PTX3 production was not increased in other alternative conditions of activation, such as in the presence of dexamethasone or calcitriol. PTX3 has properties similar to antibodies. Its production is induced by pa...

The leukapheresis product is from patients with advanced (stage III or IV) melanoma. It is used to provide sources for tumor antigens as well as monocytes and T cells, which are isolated from the leukapheresis product by elutriation.

Primed mice were treated with these antibodies. This treatment induced the development of regulatory T cells expressing high levels of ICOS and producing IL-10.

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...

A recombinant Leishmania major parasite expressing a fluorescent tracer was used for the infection of BALB/c (H2-d) mice. This was done to investigate whether infected DC process pathogen-derived antigen and stimulate antigen-specific CD4+ T cells in vivo.

2 mM L-glutamine (Sigma) was added to the monocyte cell culture.

Monocytes were isolated from PBMC to generate dendritic cells.

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.

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

Co-culture of immature DC with TIL or K562 cells overexpressing CD83 results in the production of enhanced levels of pro-inflammatory cytokines, whereas this production is less pronounced or even absent in co-cultures with non-modified TIL or K562 cells.

It has been shown that DCs express specific receptors for exosomes. Since ICAM-1 on exosomes is required for their ability to activate T cells via DCs in vitro, the role of Mac-1 and LFA-1 (the major ligands for ICAM-1) as potential receptors for exosomes has been investigated. A new role for LFA-1 on DCs, as a receptor for exosomes has been proposed.

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.