Blood samples and tumour biopsies will be taken from these newly diagnosed glioblastoma patients in order to carry out a preclinical study in which circulating tumour-specific CD8+ T-cells will be detected in their blood. In order to obtain the blood samples and tumour biopsies, a file has been submitted to the Ethics Committee of the Erasme Hospital and has been recently approved.

This patient is undergoing i.v. application of indium labelled E/L-S DC (without antigen loading) in a study of DC migration.

We have conducted a phase I long peptide vaccination trial with p53 peptides in Montanide adjuvant in patients with colorectal cancer or ovarium cancer.

We have conducted a phase I long peptide vaccination trial with p53 peptides in Montanide adjuvant in patients with colorectal cancer or ovarium cancer.

The effect of environmental factors on development of high affinity CTL was investigated using a system whereby OT-1 cells were primed in vitro by engineered APC. After a 20 h priming phase, CTL were transferred to recipient mice that were either naïve, or had been injected with activated dendritic cells one day earlier, thus creating a reactive lymph node.

Serotype A C. albicans strain SC5314 was cultured overnight in Saboraud medium at 28°C and then shifted to RPMI medium for 18 hours at 37°C to allow hyphal growth.

S. cerevisiae strain BY4741 och1 (Mata his3delta1 leu2delta0 met15delta0 ura3delta0 OCH1::kanMX4) was cultured in complete medium till exponentially phase. The cellswere biotinylated, labeled and - after treatment of DCs - detected using APC-labeled streptavidin and analyzed by flow cytometry.

S. cerevisiae strains BY4741 (genotype, Mata his3delta1 leu2delta0 met15delta0 ura3delta0) was cultured in complete medium till exponentially phase. The cells were biotinylated, labeled and - after treatment of DCs - detected using APC-labeled streptavidin and analyzed by flow cytometry.

Saccharomyces cerevisiae strain SK1 (MATa/alpha HO gal2 cupS can1R BIO, Kane SM and Roth J. 1974 Bacteriol. 118: 8-14) was cultured in complete medium (YPD, 2% yeast extract, 1% peptone, 2% glucose) for 18 hours, then collected, washed twice with sterile water and resuspended at 108 cells/ml. The cells were biotinylated, labeled and - after treatment of DCs - detected using APC-labeled streptavidin and analyzed by flow cytometry.

In our phase I clinical trial, we highlight the capacity of Dex based-vaccines to restore the number and NKG2D-dependent function of NK cells in 7/14 cancer patients.

Monocyte-derived and cocktail-matured DC electroporated with a combination of RNAs encoding tumor antigens (MelanA, Mage3, Survivin) and E/L-selectin are currently used in a clinical trial. Most patients having received the vaccine already demonstrated broad responses to numerous peptides prior vaccination. Significantly enhanced responses to several peptides and occurrence of new responses was seen in 6 patients, 5 of which were vaccinated with E/L-S+ DC. These data provide evidence for a superior immunopotency of E/L-selectin expressing DC.

Aiming at finding possible reasons for the discrepancies between our findings and published data could be the amount of ONTAK used in patients, we filed an amendment to the clinical trial allowing the inclusion of additional 4 patients. These patients will receive higher doses (12microg/kg 3x and 18microg/kg 1x) before vaccination. Taking into account direct effects of ONTAK on DC, patients will be vaccinated some days after last ONTAK treatment (as soon as blood DC have recovered to 50% of pre ONTAK amount). Third stage of DERMA-ER-DC 05 clinical trial.

In order to identify the APCs that were responsible for the development of this state of immune tolerance, we aimed to generate a transgenic mouse that express a recombinant fluorescent protein in the mammary glands, an approach that should result in high levels of expression of the fluorescent protein in the milk. We thus should be able to detect the cells that capture, degrade and present this antigen to T cells. Since we had previously generated a monoclonal antibody, 2C44, that reacts to a peptide derived from the Leishmania LACK protein bound to I-Ad MHC ...

We found that CCR6–deficient (CCR6-KO) mice were resistant to induction of EAE but became susceptible when transferred with small number of CCR6-sufficient T cells. CCR6 was found to be required on a first wave of TH-17 cells that entered the CNS through the choroid plexus epithelial cells, which constitutively expressed CCL20 in both mice and humans.

In these patients, dendritic cells were tracked for monitoring of cellular therapy by MRI. MRI cell tracking using iron oxides appears clinically safe and well suited to monitor cellular therapy in humans.

Subcutaneous injection of hypoxic DCs into the footpads of mice results in defective DC homing to draining lymph nodes, but enhanced leukocyte recruitment at the site of injection.

Tumor growth is supported by tumor stroma, such as tumor associated macrophages (TAM) and tumor associated dendritic cells (TADC). We have recently reported that TAM display massive nuclear localization of the p50 NF-kB inhibitory homodimer, which correlates with impaired inflammatory functions.The functional significance of this observation was demonstrated in p50 NF-kB deficient mice, which displayed tumor growth inhibition.

We infected mice with CD8+ T-cells primed with or without the influence of a reactive lymph node with a flu-virus encoding the specific antigen. After several days we determined the viral load in the lungs with RT-PCR. The results show that in mice with CD8+ T-cells primed with a reactive lymph node the viral load in the lungs was significantly smaller then in a control mice.

After 10’ of incubation in 65 °C water bath, the samples were incubated on ice.

The yeast cells were resuspent in 3.6 ml AE Buffer and then transferred in 15 ml tubes wit 200 µl 10% SDS (w/v). 2 ml of preheated acid phenol (pH 4.3) were added and the falcon were mixed by vortex.