We have obtained data from a phase I/II clinical trial of patients with end stage cervical cancer and of patients with vulvar intraepithelial neoplasia (VIN) grade III. Preclinical animal studies have shown that long peptides of 25-35 amino acids in length upon injection are only processed efficiently for MHC class I by dendritic cells in vivo. Long peptide injection is followed by appropriate cognate interactions between such DC, CD4+ T helper cells and CD8+ CTL precursors. Three groups of end stage cervical cancer patients (in total N=35) were subcutaneously va...

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 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.

Design of this trial: sequential adaptive design, 2nd cohort, mature DC transfected with RNA encoding MelanA, Mage3 and Survivin +/- E/L-selectin, i.v. route, 13 evaluated patients. Comparison of pre-existing and vaccine-induced immune responses in stage IV melanoma patients. We have efficiently transfected DC with RNA encoding a functional protein (E/L-selectin), which allows entry of DC into LN from HEV. These DC rolled in vitro on sialyl-LewisX-coated slides, and in vivo, mouse E/L-selectin-transfected DC homed to LN after i.v. application. Monocyte-derived and...

Our lab has data from an interim statistical analysis of the DERMA-ER-DC 04 trial (multipeptide loaded cytokine matured moDC +/- CD40L activation, 70 evaluated patients). This analysis for the first time showed a clinical correlation of induced immune responses as measured in the blood with outcome, as there have been so far statistically significantly less events in stage III patients showing good CTL reponses (IFN-y Elispot) to multiple class I and II peptides compared to low responders of the stage III cohort. Additional in vitro maturation of DC by CD40L (leuci...

We have data available on the induction of effective therapeutic antitumor immunity by direct in vivo administration of ovalbumin (OVA) encoding lentiviral vectors. Ex vivo lentivirally transduced dendritic cells (DC) have been described to induce CD8+ and CD4+ T-cell responses against various tumor-associated antigens (TAAs) in vitro and in vivo. We report here that direct administration of ovalbumin (OVA) encoding lentiviral vectors caused in vivo transduction of cells that were found in draining lymph nodes (LNs) and induced potent anti-OVA cytotoxic T cells ...

Data were obtained on responses to viral infections in TLR9 -/- mice, C57Bl/6 mice and IFNalpha receptor -/- mice. Toll receptor 9 (TLR9) is an important pattern recognition receptor. It is stimulated by foreign DNA or CpG motivs. We found that TLR9 -/- mice were very susceptible to infection with the mouse pox virus Ectromelia. In contrast to DC from C57Bl/6 mice, DC from TLR9 -/- produced little if any interferon alpha. However, MVA-BN induced IFNalpha in presence or absence of TLR9 -/-. Moreover, coinfection with MVA-BN and Ectromilia protected TLR9 -/- mice ...

Our lab gathered data on the responses to EBV and HIV in hu-SCID mice.

We have obtained data from an analysis of the uptake, conservation and cross-presentation of the model antigen OVA after Fc receptor-mediated uptake by DC and from an analysis of the immunological consequences of the antigen depot. We found that MHC class I presentation is relatively short-lived in contrast to MHC class II. However, CD8 cross-priming capacity of OVA-loaded DC was functionally retained for many days, while peptide-pulsed DC had lost their priming capacity after 24 hours. Strikingly, OVA protein antigen was conserved intracellularly in DC for many ...

Our laboratory obtained data on the induction of immune responses after targeting antigens to DC via TLR agonists.

Our lab has obtained data on the mechanism of how IL-10 regulates the immune response to MTb infection in mice. Control and clearance of intracellular pathogens such as MTb is dependent on the production of TNF and the induction of the T-helper 1 (Th1) cytokine IFN-gamma by IL-12. Conversely, IL-10 is a suppressive cytokine essential for dampening the immune response to a number of intracellular pathogens to limit host immune pathology. IL-10 has been shown to exert its suppressive effect by directly acting on the antigen presenting cell (APC), therefore functi...

Our lab has gathered data in establishing a per-clinical model of breast cancer.

In our lab, data was obtained from a phase I trial using peptide-pulsed Dex. Dendritic cell (DC) derived-exosomes (Dex) are nanomeric vesicles harboring functional MHC/peptide complexes promoting T cell -dependent tumor rejection. In this Phase I trial using peptide-pulsed Dex, the observation of clinical regressions in the absence of T cell responses prompted the search for alternate effector mechanisms. Mouse studies unraveled the bioactivity of Dex onto NK cells, Dex promoting a IL-15Ra-dependent NK cell proliferation and a NKG2D-dependent activation resulting...

We obtained data from studies of CD8 T cell properties in man comparing HIV, CMV and EBV-specific cells. It is usually assumed that effector gene expression in CD8 T cells may be similar in all immune responses, CD8 T cells in vivo differentiating into cytotoxic T cells that also produce g-inf. In contrast we published in 2007 that CD8 follow several differentiation waves and that effector gene expression is heterogeneous, different effector genes having different kinetics of expression-down regulation and associating randomly, suggesting that different immune re...

We obtained data on immune tolerance in mice breast-fed by antigen-exposed lactating mothers. We have previously generated a monoclonal antibody, 2C44, that reacted to a peptide derived from the Leishmania LACK protein bound to I-Ad MHC class II molecules. We have successfully used the 2C44 mAb to visualize LACK-presenting DCs in mice infected with the L. major. In 2007, we have attempted to identify the APCs that are responsible for the development of immune tolerance in mice that have been breast-fed by antigen-exposed lactating mothers. Recent experiments perf...

Our laboratory obtained data from studies of DC phagosomes and cross-presentation using K/O and mutant mouse strains. Cross presentation is the process by which Dendritic cells (DC) phagocytose pathogens or dying cell fragments, and present proteolytic peptides derived from these antigens in association with MHC class I molecules. The reasons why DCs are the only antigen presenting cells that efficiently cross present antigens are not well understood. We have demonstrated that the NADPH oxidase NOX2 is recruited to DC early phagosomes mediating a sustained produc...

We obtained data from an evaluation whether antigens could be rerouted from the endosomal into the cytosolic compartment by combining antigen with cell penetrating peptides to further enhance cross-presentation. We made significant progress using biodegradable polymers to enhance cross-presentation of loaded antigens, and the best results were obtained by maturing DCs with a combination of the TLR ligands R848 and polyI:C. Furthermore, we have shown that DC electroporated with melanoma antigen RNA express the proteins ex vivo and in vivo.

Our lab obtained data on maturation of DC by TLR agonists.

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.

Data have been obtained on the use of nano-particles for intestinal vaccine delivery from our continued investigation of the use of nano-particles as oral vaccine vehicles (Cerovic). We have identified an important role for complement C1q in the uptake of scrapie agent by dendritic cells (Flores-Langarica).We have shown that a protein from Schistosomes has dramatic effects on intestinal physiology, including dendritic cells (Nassar). We have shown that steady-state intestinal denritic cells are not constitutively biased towards stimluating Th2 responses (Milling). ...