77 patients have been enrolled in two Phase III trials from 2 institutions testing dosing or duration of IM administration (EORTC 62005 and BFR14 trials).

A functional analysis of anti-MAGE-3.A1 CTL clones derived from vaccinated patients (either ALVAC canarypox virus vector, peptide-pulsed DC or peptide vaccination +/- montanide adjuvant) who displayed tumor regression was done. The functional avidities of these CTL clones, evaluated in lysis assays, were surprisingly low, suggesting that high avidity was not part of the putative capability of these CTL to trigger tumor rejection. Most anti-MAGE-3.A1 CTL clones obtained after DC vaccination, but not after peptide or ALVAC vaccination, produced IL-10. Transcri...

These DC pulsed with MAGE-A3, gp100, NA17 and tyrosinase were used for vaccination of melanoma patients.

The melanoma patients were vaccinated with a MAGE-A3 peptide presented by HLA-A1 without adjuvant, while other melanoma patients were vaccinated with ALVAC canarypox virus containing a MAGE-A3 minigene, or peptide-pulsed dendritic cells (DC). There was a correlation between tumor regression and detection of anti-MAGE-3.A1 CTL responses.

The patients were vaccinated with 4 peptides (MAGE-A3, gp100, NA17 and tyrosinase) presented by HLA-A2. We measured responses to these peptides injected with Montanide, but did not know whether the Montanide adjuvant was important for these T cell responses. Indeed, melanoma patients may mount spontaneous responses to these antigenic peptides. We therefore injected the same peptides without adjuvant, and no response was observed. We conclude that Montanide had a clear adjuvant effect for the CD8 T cell responses measured against these four peptides.

Post-vaccination frequencies of the T cells of anti-NY-ESO-1.A2 and anti-Tyrosinase.A2 were found to be >= 10-fold higher than that of pre-vaccinations, indicating a specific CTL response to these vaccinations. An analysis performed on PBL collected after the 3 vaccinations of cycle 2 clearly shows that the frequencies for both anti-NY-ESO-1.A2 and anti-Tyrosinase.A2 T cells are still elevated.

Post-vaccination frequencies of the T cells of anti-NY-ESO-1.A2 and anti-Tyrosinase.A2 were found to be >= 10-fold higher than that of pre-vaccinations, indicating a specific CTL response to these vaccinations. An analysis performed on PBL collected after the 3 vaccinations of cycle 2 clearly shows that the frequencies for both anti-NY-ESO-1.A2 and anti-Tyrosinase.A2 T cells are still elevated.

Tests were performed on frozen peripheral blood leukocytes (PBL) obtained from leukapheresis or buffy-coat collections performed before and after the 6 vaccinations in Cycle 1 and after the 3 vaccinations of Cycle 2. The fresh PBL samples were collected from UTCM (Hôpital Erasme, ULB) and were transferred to Ludwig Institute where the freezing of the samples and analyses were performed.

A vaccination trial with RNA transfected autologous DC was done with overlapping peptides covering the whole protein sequence of the tumor antigens MageA3, MelanA and Survivin. With this approach we monitored the immune responses of 8 vaccinated patients and of more than 15 other stage IV melanoma patients.

These multimers were provided by P. Coulie and used to establish routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry.

The cDNA’s encoding these early expressed HIV antigens have been human codon optimized and modified with lysosomal targeting sequences.

An HIV infected individuals stable under HAART study has been initiated. These patients are being vaccinated with cytokine cocktail matured DCs electroporated with mRNA encoding Tat, Rev and Nef. The cDNA’s encoding these early expressed HIV antigens have been human codon optimized and modified with lysosomal targeting sequences. So far, 17 patients have been included in the study.

These patients were treated with monocyte-derived DCs matured with inflammatory cytokines and subsequently electroporated with mRNA encoding 6 tumor-associated antigens.

We investigated the interactions between human monocyte derived DCs (MDDC), generated in the presence of GM-CSF and IL-4 (IL-4 DC), and antigen-stimulated circulating gamma delta T lymphocytes, bearing the Vgamma2 TCR.

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 similar to those elicited by ex vivo transduced DC.

These mice were studied with regard to responses to viral infections and compared to TLR K/O mice. We used the mouse pox virus Ectromelia and MVA-BN.

These K/O mice were studied with regard to responses to viral infections. We used the mouse pox virus Ectromelia and MVA-BN.

We have analyzed the uptake, conservation and cross-presentation of the model antigen OVA after Fc receptor-mediated uptake by DC. 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.

These monocyte-derived DCs received combined stimulation with TLR ligands. They were used in our study on DC activation by pathogen-derived stimuli.

The question was addressed of whether bLf, whose immunomodulant properties are now recognized, could influence MDDC differentiation/activation.