These cells are incubated in Fc-block (1:50 in PBS + 2 % FCS) 30 min on ice, to be stained for FACS.

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 analysed the ability of invariant NKT (iNKT) cells to assist priming of antigen specific T and B cell responses. 1) 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. 2) 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. Using a soluble iNKT TCR and a newly engineered anti...

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.

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...

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.

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).

A long-standing collaboration with Centre for Ecology & Hydrology (CEH, UK) has led to the identification of an activity in salivary gland extracts of the tick Rhipicephalus appendiculatus that profoundly modulates the responses of human monocyte-DC towards extrinsic stimuli. The gene for this molecule, which we have termed Japanin, has been cloned and recombinant protein has been generated. This material inhibits the up-regulation of costimulatory molecules in response to certain TLR agonists, and some cytokines but not others, and modulates the function of...

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.

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.

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

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

Tetanus toxide epitope embedded into a L-SIGN/DC-SIGN-cross-reactive Ab was targeted to dendritic cells. We believe that the isolated Abs may be useful for selective delivery of Ags to DC-SIGN- or L-SIGN-bearing APCs for the modulation of immune responses and for blocking viral infections.

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

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.

There are lentiviral vectors available in the lab.

The Leukapheresis product from cancer patients was used to produce enriched monocytes using elutriation.