We have investigated the respective role of IL-10 and IDO, both required for Th1 suppression in this model. As IL-10 has been shown to indirectly dampen Th1 responses through the inhibition of IL-12 production by antigen-presenting-cells, we monitored mRNA expression specific for the inducible chain IL-12 p35 in lymph nodes draining the site of injection or in mesenteric lymph nodes of mice instilled intra-colonically with TNBS. Our results clearly show that IL-12 production early after the onset of the response was unaffected by anti-CTLA-4 treatment. By cont...

It was shown that both IL-10 as well as IL-12 production is dependent on the signalling adaptor molecules either MyD88 or TRIF, respectively in response to CpG, LPS or PolyIC.

It was shown that both IL-10 as well as IL-12 production is dependent on the signalling adaptor molecules either MyD88 or TRIF, respectively in response to CpG, LPS or PolyIC.

Monocytes were transferred into culture bags by sterile welding and cultured for 5 days in serum free medium (CellGro, Cell Genix) in the presence of 100 ng/mL GM-CSF (Leukine, Berlex) and 20 ng/mL IL-4 (Cell Genix).

ImmunoVexHSV2 is a product for the prevention of infectious disease, it is a vaccine for genital herpes. In preclinical studies, ImmunoVex HSV2 completely prevents disease and invokes a powerful immune response.

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.

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...

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

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.

There are lentiviral vectors available in the lab.

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.

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...

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.

This vaccine is composed of DC pulsed with 2 to 3 MAA-derived peptides (combinations of MAGE-3.A2, MAGE-C2.A2 or Na17.A2). It was tested in a trial in small cohorts of patients (3-9) who were vaccinated with differently composed DC-vaccines.

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.

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

Mannan (500 micrograms/ml) was added to DCs in order to block their mannan receptors.