One volume fresh medium containing 2x concentrated cytokines was added to the cell culture of monocytes in serum free medium in the presence of 100 ng/mL GM-CSF (Leukine, Berlex) and 20 ng/mL IL-4 (Cell Genix).

When identifying molecular signatures for alternatively activated DC (AA-DC) it was found that the hallmark of AA-DC was the production of the angiogenic cytokine VEGF in vitro, and in vivo when the cells were implanted into the chicken embryo CAM assay. VEGF production by DC was selectively observed only when the DC where alternatively activated. Therefore, VEGF production by DC can be considered a signature of this state of activation.

APC-labeled streptavidin was used to detect intracellular yeasts/spores.

These surface markers were used to establishe routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry. For this purpose multimers were used kindly provided by Pierre Coulie (Brussels) together with surface markers (such as CD8, CD4, CD3, CD45RA, CCR7, CD25, CD27, CD137, CD127) in different 8-color panels on the BD FACSCanto II. Additionally, functional markers such as CD107a were used and intracellular cytokine staining for the further analysis of individual T-cell populations and T-cell clones.

CD1 molecules that are subsequently to be refolded in vitro in order to generate CD1 tetramers.

CD1 molecules are refolded in vitro in order to generate CD1 tetramers.

Surface markers were used to establishe routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry. For this purpose multimers were used kindly provided by Pierre Coulie (Brussels) together with surface markers (such as CD8, CD4, CD3, CD45RA, CCR7, CD25, CD27, CD137, CD127) in different 8-color panels on the BD FACSCanto II. Additionally, these CD107a functional markers were used and intracellular cytokine staining for the further analysis of individual T-cell populations and T-cell clones.

Surface markers were used to establishe routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry. For this purpose multimers were used kindly provided by Pierre Coulie (Brussels) together with CD127 surface markers (and additionally, CD8, CD4, CD3, CD45RA, CCR7, CD25, CD27, CD137) in different 8-color panels on the BD FACSCanto II. Additionally, functional markers such as CD107a were used and intracellular cytokine staining for the further analysis of individual T-cell populations and T-cell clones.

Surface markers were used to establishe routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry. For this purpose multimers were used kindly provided by Pierre Coulie (Brussels) together with CD137 surface markers (and additionally, CD8, CD4, CD3, CD45RA, CCR7, CD25, CD27, CD127) in different 8-color panels on the BD FACSCanto II. Additionally, functional markers such as CD107a were used and intracellular cytokine staining for the further analysis of individual T-cell populations and T-cell clones.

The CD1d tetramers were generated from in vitro refolded CD1 molecules. The ability to generate CD1d tetramers has provided us with the opportunity of comparing a broad panel of CD1d binding compounds for their ability to stimulate iNKT cells.

Surface markers were used to establishe routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry. For this purpose multimers were used kindly provided by Pierre Coulie (Brussels) together with CD25 surface markers (and additionally, CD8, CD4, CD3, CD45RA, CCR7, CD27, CD137, CD127) in different 8-color panels on the BD FACSCanto II. Additionally, functional markers such as CD107a were used and intracellular cytokine staining for the further analysis of individual T-cell populations and T-cell clones.

Surface markers were used to establishe routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry. For this purpose multimers were used kindly provided by Pierre Coulie (Brussels) together with CD27 surface markers (and additionally, CD8, CD4, CD3, CD45RA, CCR7, CD25, CD137, CD127) in different 8-color panels on the BD FACSCanto II. Additionally, functional markers such as CD107a were used and intracellular cytokine staining for the further analysis of individual T-cell populations and T-cell clones.

Surface markers were used to establishe routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry. For this purpose multimers were used kindly provided by Pierre Coulie (Brussels) together with CD3 surface markers (and additionally, CD8, CD4, CD45RA, CCR7, CD25, CD27, CD137, CD127) in different 8-color panels on the BD FACSCanto II. Additionally, functional markers such as CD107a were used and intracellular cytokine staining for the further analysis of individual T-cell populations and T-cell clones.

Surface markers were used to establishe routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry. For this purpose multimers were used kindly provided by Pierre Coulie (Brussels) together with CD4 surface markers (and additionally, CD8, CD3, CD45RA, CCR7, CD25, CD27, CD137, CD127) in different 8-color panels on the BD FACSCanto II. Additionally, functional markers such as CD107a were used and intracellular cytokine staining for the further analysis of individual T-cell populations and T-cell clones.

Long peptides which were covalently conjugated to either the TLR2 ligand or TLR9 ligand, Pam3CysSK4 or CpG have been used to study the uptake, antigen presentation, and induction of specific T-cells.

Constructs for fusion proteins of the DC specific receptor DC-SIGN fused to GFP have been completed and tested.

The model antigen OVA was fused to the C1C2 exosome-binding domain of MFG-E8/lactadherin. This construct was used in a study on the capture of antigen-bearing exosomes by DCs and cross-presentation of tumour antigen in exosomes.

Fabs that bind strongly to L-SIGN, but to a lesser degree or not at all to dendritic cell-specific ICAM-grabbing nonintegrin (DC-SIGN) were isolated and characterized. 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.

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).

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.