Total RNA was extracted from yeast cells after buffering, incubation and centrifugation by phenol extraction.

DMSO was used to dissolve blue formazan particles.

Three patients were enrolled in a clinical trial and DC vaccine was produced. The DC vaccine was administered into irradiated tumours in RCC of one patient only due to cancer progression in the other two patients. The patient treated is in complete remission 6 months after treatment.

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

This DC vaccine was produced from the Leukapheresis product, after elutriation, of a metastatic lesion of stage III/IV melanoma patients.

This DC vaccine was produced from matured DC (matured using TNF alpha, IFN alpha and PGE2) which were subsequently transfected with CD40L-encoding RNA and tumor RNA.

Autologous dendritic cells are loaded with apoptotic leukemic cells from patients with B-CLL to be used as vaccine in patients with chronic lymphocytic leukemia. Preliminary data from a clinical study indicate reduction of circulating tumor cells. No adverse effects have been observed.

This vaccine was used for RCC patients who were deficient in T cell IFN-gamma and IL-2 production pre-vaccination.

Using curdlan as a specific agonist of dectin-1, we have shown that this C-type lectin couple to Syk kinase leads to activation of ERK, JNK and p38 MAPKs, as well as NF-kappaB.

These constructs were used to transfect immature DCs.

DC based vaccines expressing the tumor antigens PSA or Her2/neu were produced in a pre-clinical work.

100 micrograms/ml curdlan (Wako) was used (along with R848, LPS, yeast RNA and yeast cells in different conditions of culture) to induce DC activation.

We have prepared several new constructs for the expression of (modified) adhesion receptors in DC.

We have prepared several new constructs for the expression of c-type lectin receptors in DC.

DCs were exposed to cytochalasin D (10 microg/ml, TebuBio) for 30 minutes at 4°C.

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.

CLL-DCV01 is used for multiple injections in patients with previously treated B-cell Chronic Lymphocytic Leukemia.

This RNA was used to transfect matured DC with the final aim to produce a DC vaccine.

Surface markers were used to establish 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 CD45RA surface markers (and additionally, CD8, CD4, CD3, 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 establish 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 CD8 surface markers (and additionally, 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.