We attempted to improve the adoptive transfer protocol for immunotherapy by stimulating T cells with monocyte derived DC pulsed with tumor lysate, instead of simply adding tumor lysate into PBMC cultures. T cells raised exhibited some tumor reactivity and outgrowth of a distinct T cell population could be observed.

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.

We assessed the capacity of the electroporated DCs to activate naive HLA-A2-restricted MelanA-specific CD8(+) T cells without the addition of any exogenous cytokines. A >500-fold increase in MelanA-specific CD8(+) T cells was observed when compared with immature DCs, and a >200-fold increase when compared with cytokine cocktail-matured DCs. In correlation, we found a marked increase in cytolytic and IFN-gamma/tumor necrosis factor-alpha (TNF-alpha) secreting CD8(+) T cells. Our data indicate that immature DCs genetically modified to express stimulating molecul...

These cells do not acquire a mature phenotype and do not lead to an enhanced secretion of several cytokines/chemokines.

These cells acquire a mature phenotype along with an enhanced secretion of several cytokines/chemokines. Moreover, these DCs are very potent in inducing naive CD4(+) T cells to differentiate into interferon-gamma (IFN-gamma)-secreting type 1 T helper (Th1) cells.

We have shown recently that the T cell stimulatory capacity of DCs pulsed with tumorantigen-derived peptides can be considerably increased by activating the DCs through electroporation with CD40L, CD70 and constitutively active TLR4 encoding mRNA (TriMix DCs). This vaccine will also be tested in the multicentre trial.

Attenuated Expression of A20 Markedly Increases the Efficacy of Double Stranded RNA-Activated Dendritic Cells as an Anti-Cancer Vaccine A20 is a zinc finger protein with ubiquitine-modifying activity. It has been described that A20 negatively regulates signaling induced by the tumor necrosis factor receptor and toll like receptor (TLR) family in a number of cell types, including mouse bone marrow-derived dendritic cells (DCs). However, the expression and effect of A20 in activated monocyte-derived DCs have not been previously evaluated. We report that DC...

We established tumor cell lines from renal cell cancer in our laboratory.

We established tumor cell lines from pancreatic cancer in our laboratory.

We established tumor cell lines from melanoma in our laboratory.

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.

We have chemically coupled the S-epimer of the Pam3CSK4 to long peptides containing a CTL epitope and used this structure to investigate the behaviour of the diastereomer.

We have chemically coupled the R-epimer of the Pam3CSK4 to long peptides containing a CTL epitope and used this structure to investigate the behaviour of the diastereomer.

By using TAP deficient cells, we found that TAP is also necessary for the process of presentation of the antigen in MHC class I. We further characterized the antigen depot by confocal microscopy and found that the antigen co-localized with the lysosomal marker LAMP1, but not with the endosomal marker EEA1, TAP, MHC class I or MHC class II. We conclude that the antigen depot is a storage compartment and not a loading compartment. We have started to examine depot formation by targeting antigen to other receptors. For example, we have found that presentation ...

We have prepared two plasmids coding for LACK linked to either eGFP or mCherry fluorescent proteins, under the control of the beta-casein promoter. 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 cel...

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.

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

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