Our lab obtained data on the kinetics of effector gene expression and association in TCR-Tg CD8 cells responding to the male antigen and to Listeria-OVA with regard to « in vivo » responses to Ags. Surprisingly, instead of a single program of differentiation leading to the coordinated expression of all effector genes; we found that individual effector genes had different kinetics of expression and associated randomly generating several new effector cell types. These results are published in J. Exp. Med. Since these results were obtained in TCR-Tg mice, and thus ...

Our lead product for the treatment of cancer is OncoVex GMCSF. OncoVexGMCSF is completing a 50-patient Phase II clinical trial for the treatment of unresectable metastatic melanoma, as a monotherapy, in the second line/salvage therapy setting. Data so far indicates that OncoVex can destroy both injected and un-injected melanoma deposits. The trial is still in progress.

At the end of 2004 BruCells has initiated two identical clinical studies entitled: “A Phase IB/II Study of Immunotherapy with Autologous Dendritic Cells Pulsed with Multiple HLA-A2 and MAGE-3.DP4 Peptides in Metastatic Cutaneous Melanoma Patients”. Patients received sequential immunizations with autologous dendritic cells pulsed with 8 HLA-restricted HLA-A2 peptides and a MAGE-3.DP4 peptide. They were randomized to receive immunizations either with I3 DC (DC generated with interferon-beta and interleukin-3) or with G4 DC (DC generated with GM-CSF and IL-4). At ...

Our lab gathered data on DC activation by pathogen-derived stimuli. We focused on the study of the response of MDDCs to the combined stimulation with TLR ligands and we observed that simultaneous activation of TLR4 and TLR8 signaling cascades results in a marked inhibition of the secretion of the proinflammatory chemokine CCL2 with respect to stimulation through a single TLR. This inhibition is specific for both CCL2 and TLR agonist combination and could represent a novel regulatory mechanism evolved to maintain immunological balance (Del Cornò et al.). Future st...

My laboratory obtained data from a continued analysis of the ability of invariant NKT (iNKT) cells to assist priming of antigen specific T and B cell responses. We have carried out three complementary lines of research: 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 g...

Our lab obtained data on the activation of immature monocyte-derived dendritic cells after transduction with high doses of lentiviral vectors. Dendritic cells (DCs) are an attractive tool for immunomodulation, targeting mature DCs (mDCs) for immunization or immature/semimature DCs (iDCs) for tolerization. Therefore, introducing antigens into DCs has become a prime topic in various immunological disciplines. Numerous studies have shown that lentiviruses are an efficient vehicle for this purpose. This study evaluates the effects of lentiviral transduction on iDC ...

We have obtained data from in vitro studies to demonstrate that DC can be activated via the dectin-1/Syk pathway to become APC capable of priming CD8+ T cells. In addition, we have shown that the same DC can interact with Tregs and convert them into cells that co-express ROR?t and Foxp3 and produce IL-17. We have shown that this pathway can serve as an alternative to TLR signalling for allowing DC to become effector cells capable of priming CD4+ T cells. We have used agonists of the dectin-1/Syk pathway as adjuvants in vivo to induce DC activation and CD4+ T cell ...

Our lab has data available on the effect of endogenous IL-10 production by DC subsets resulting in low levels of IL-12p70 production in mice. We have shown that plasmacytoid pDC respond to both CpG and the TLR-7 ligand R848 to produce very high levels of IL-12p70 (in the ng/ml range per 5 x 105 cells/ml) and IFN-gamma. However, BM-myeloid and splenic CD8alpha+ DC produced at least fivefold less IL-12p70, and CD8alpha- DC produced undetectable levels of IL-12p70 upon stimulation with CpG. We show that induction of endogenous IL-10 production by these latter DC ...

Our lab generated data on alternatives to the ‘classical’ maturation method (i.e. a cocktail of inflammatory cytokines) for moDC (e.g. ‘clinical grade reagents such as Ampligen TM and Hiltonol TM). Moreover the possibility to co-electroporate moDC with constitutively active TLRs and TAA encoding mRNA was persued. The in vitro charging of monocyte derived DC (moDC) with antigens has been optimised. In contrast to previous studies, the electroporation of TAA encoding mRNA after in vitro maturation of the moDC resulted in a better electroporation efficiency an...

We have data available from an analysis of the proteome of the phagocytic/endocytic compartments that are formed upon cell entry of antigens via specific pathways. The ultimate goal is to get a better insight into the protein complexes that are involved in antigen processing, and learn whether the different entry routes for antigens into DCs might have distinct functional characteristics.

We found that Indium111 labelled DC should not be loaded with tumor antigens if applied in patients, which may result in loss of specific immune responses and are testing this in-vivo. Analysis of DC migration in vivo in a first patient by i.v. application of indium labelled E/L-S DC (without antigen loading) is scheduled for March 2009. Data will become available.

We have used fluorescence microscopy to analyse the endocytic pathway in DC and to track C-type lectins that are internalised from the cell surface. From these analyses, there is data available. As part of this work, we have used microscopy to study the effect of Dectin-1 uptake on its ability to signal for downstream responses. Finally, we have shown that fluorescently-labelled antibodies against a novel C-type lectin, DNGR-1, can be used to mark DC upon injection into mice and might therefore be useful for in vivo imaging studies. Microscopy will continue to b...

We have generated data on the kinetic of the transcriptional profiling of D1 cells treated with either DMSO or LPS.

We have data on the kinetic of the transcriptional profiling of human MoDC treated with IL2 in vitro.

Our laboratory has obtained data on the kinetic of the transcriptional profiling of purified mouse DC treated with IL10.

A large reference ‘golden experiment’ will be carried out in which the transcriptome of DC vaccines of 50-100 patients will be compared and data transcriptomic data will become available.

We annotated DC specific pathways and integrated them in the pathway tool. Initially we have annotated DC specific pathways using information publically available on the web. A preliminary curated non exhaustive pathway list can be found in the DC-THERA version of the program DC_Eu.Gene.

Data has been obtained in a study of antigen recognition of blood-derived T cells. In this study, an IFNgamma ELISPOT was done with PBMC loaded with peptide pools, each consisting of 10 15-mers, which overlap with 11 aminoacids.

Data have been obtained on the antigen recognition of DTH infiltrating T cells. To obtain the data, activation of CD4 and CD8 T cells upon recognition of EBV-B cells expressing vaccinal antigens was evaluated by CD137 upregulation, CD107a and CD40L expression using flow cytometry. Luminex technology was used to quantify the TNFalpha and IFNgamma secretion.

We have obtained data from an analysis of antigen loading of DCs via targeting DC-SIGN, which resulted in enhanced class II-mediated presentation. Constructs for fusion proteins of the DC specific receptor DC-SIGN fused to GFP have been generated.