Differentiation of monocytes into dendritic cells is promoted, along with recombinant IL-4, by addition of granulocyte-macrophage colony-stimulating factor (GMCSF 1000U/ml, Chemicon).

HLA-DP4/MAGE3 molecules have been prepared to stain ex vivo PBL from immunized melanoma patients for a study of anti-tumor responses in melanoma patients that were vaccinated with autologous DCs loaded with a combination of tumor Ag peptides.

Patients with resected HPV16-positive cervical cancer were vaccinated with an overlapping set of long peptides comprising the sequences of the HPV16 E6 and E7 oncoproteins emulsified in Montanide ISA-51.

Patients with resected HPV16-positive cervical cancer were vaccinated with an overlapping set of long peptides comprising the sequences of the HPV16 E6 and E7 oncoproteins emulsified in Montanide ISA-51.

IFN alpha was used in a DC maturation process with the final aim to produce a DC vaccine.

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).

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...

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.

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...

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.

Soluble recombinant HLADR1101 MHC class II molecules receptive for loading with either pathogen or tumor-derived peptides were constructed for a study of CD4+ T cell responses against pathogens or tumors in humans.

An mAb reacting to the immunodominant LACK156-173 peptide of leishmania bound to I-Ad MHC class II molecules was prepared. To this aim, we injected I-Ad/LACK recombinant dimers to TCR transgenic mice which exhibited an increased frequency of LACK-specific T cells. Four out of 600 supernatants, i.e. 2C44, 2F74, 2E60 and 2X8, readily stained LACK156-173-pulsed fibroblasts, but neither unpulsed fibroblasts nor OVA323-339-pulsed cells. BIAcore measurements yielded equilibrium dissociation constants ranging from 1.1 nM for 2C44 mAb to 120 nM for 2F74 mAb.

We have generated a monoclonal antibody, 2C44, that reacted to a peptide derived from the Leishmania LACK protein bound to I-Ad MHC class II molecules. We have successfully used the 2C44 mAb to visualize LACK-presenting DCs in mice infected with the L. major.

The expression and function of these murine FcgammaR in CD11c+CD11b-B220+ plasmacytoid DCs (pDCs) was investigated. pDCs express mostly FcgammaRIIB while the expression of FcgammaRI and FcgammaRIII is only detected by RT-PCR at low but significant level. Moreover, the ITAM-containing intracellular chain associated to FcgammaRI and FcgammaRIII is strongly expressed in pDCs as detected by biochemical assay.

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, but did not know 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.