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.

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.

DC-based vaccines that were given to these patients were combined with low dose IFN-alfa (3 x 10e6 U per wk). In a number of patients we observed vaccine-induced depigmentation and vitiligo.

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.

Melanoma patients were vaccinated with peptide-pulsed DC (MAGE-A3, gp100, NA17 and tyrosinase), while other patients were vaccinated with peptides +/s adjuvant or viral vectors (ALVAC canarypox virus containing a MAGE-A3 minigene). There was a correlation between tumor regression and detection of anti-MAGE-3.A1 CTL responses.

Melanoma patients were administered ALVAC canarypox virus containing a MAGE-A3 minigene, while others were vaccinated with MAGE-A3 peptide presented by HLA-A1, administered as peptide, and peptide-pulsed DCs. There was a correlation between tumor regression and detection of anti-MAGE-3.A1 CTL responses.

Patients with stage III or stage IV melanoma who undergo surgical resection of a melanotic lesion. These patients are vaccinated with pulsed and thouroughly tested dendritic cell vaccine.

Small cohorts of patients (3-9) were vaccinated with differently composed DC-vaccines: DC loaded with a single MAA-derived peptide (MAGE-3.A1 or -A2 or Na17.A2) or pulsed with 2 to 3 MAA-derived peptides (combinations of MAGE-3.A2, MAGE-C2.A2 or Na17.A2) or G4-DC loaded with 8 different peptides. An important conclusion that can be made from these pilot-observations is that the potential therapeutic effect of our DC-based MAA-vaccination approach does not occur instantly but is delayed by 8-12w following the first vaccination (i.e.12-16w after the isolation o...

A significant number of patients was enrolled in the DC-based immunotherapy trial. After vaccination, vaccine-induced depigmentation and vitiligo were observed.

Anti-tumor responses were followed in melanoma patients that were vaccinated with autologous DCs loaded with a combination of tumor Ag peptides.

This HSV vector expressing full length tyrosinase, gp100 and MART-1 was used to transduce dendritic cells from melanoma patients which were then to be used for vaccination.

This patient experienced an outstanding response to immunotherapy. The patient received several adoptive T cell transfers and regressed from stage IV melanoma to a disease free state and has remained tumor free for several years.

Antigen-specific CD4+ T cells were followed in these immunized individuals using generated peptide-MHC class II multimers.

This patient has received 9 vaccines with peptides only in a randomized trial 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). Up to now, in Cycle 3, immunizations have been continued with peptide-only injections of all 9 peptides except Tyrosinase at 12 weeks intervals. For the continuation of the vaccinations the clinicians have decided to inject the patient at 6 months intervals. The patient who clinically showed Partial Response at the end of cycle 1 is curre...

These patients undergo leukapheresis and later intradermal administration of the produced vaccine (V administrations of autologous dendritic cells generated from adherent peripheral blood monocytes and subject to quality controls - with eventual additional ones depending on clinical outcome).

The patients are immunized with autologous DC loaded with KLH, as immunological tracer, and an allogeneic peptidome (i.e. natural tumour peptides, NTPs), obtained from melanoma cell line SK-Mel24.

An HLA-A2 positive healthy donor underwent leukapheresis with the aim to generate monocyte derived DC under GMP conditions.

These patients undergo leukapheresis and later intradermal administration of the produced vaccine (V administrations of autologous dendritic cells generated from adherent peripheral blood monocytes and subject to quality controls - with eventual additional ones depending on clinical outcome).

The patients are immunized with autologous DC loaded with KLH, as immunological tracer, and an allogeneic peptidome (i.e. natural tumour peptides, NTPs), obtained from melanoma cell line SK-Mel24.

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.