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...

Four patients are enrolled in a clinical trial for vaccination with DC loaded with apoptotic leukemic cells. A strong type I T cell response has been induced. Preliminary data also indicate reduction of circulating tumor cells. No adverse effects have been observed.

Autologous dendritic cells are loaded with apoptotic leukemic cells from patients with B-CLL to be used as vaccine in patients with chronic lymphocytic leukemia. Preliminary data from a clinical study indicate reduction of circulating tumor cells. No adverse effects have been observed.

Three patients were enrolled in a clinical trial and DC vaccine was produced. The DC vaccine was administered into irradiated tumours in RCC of one patient only due to cancer progression in the other two patients. The patient treated is in complete remission 6 months after treatment.

Three patients were enrolled in a clinical trial of a DC vaccine administered into irradiated tumours in RCC. However only this one patient received the vaccine due to cancer progression in the other two patients. The DC were labelled with 111In. The following patients will receive DC labelled with Endorem and the vaccine will be imaged with MRI. The patient treated is in complete remission 6 months after treatment.

HLA-DP4/MAGE3 molecules have been prepared to stain ex vivo PBL from immunized melanoma patients for a study of anti-tumor responses in melanoma patients that were vaccinated with autologous DCs loaded with a combination of tumor Ag peptides.

Soluble recombinant HLADR1101 MHC class II molecules receptive for loading with either pathogen or tumor-derived peptides were constructed for a study of CD4+ T cell responses against pathogens or tumors in humans.

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).

We have attempted to identify the antigen-presenting cells (APCs) that are responsible for the development of immune tolerance in mice that have been breast-fed by antigen-exposed lactating mothers. Recent experiments performed in our laboratory have shown that the exposure of a mother to an airborne antigen during lactation impacts asthma development in their progeny. We found that airborne allergens were efficiently transferred from the mother to the neonate through the milk and that this transfer resulted in the development of antigen-specific tolerance.

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.

We set an in vitro system to study ex-vivo responses able to detect fona fide in vivo primed CD4+ T cells. We investigated spontaneous CD4+ T cell responses to these antigens in normal donors and in patients with high-grade cervical lesions, pancreas adenocarcinoma and advanced melanoma. The antigens were: i) the E6 and E7 proteins of human papilloma viruses, ii) the carcinoembryonic antigen (CEA) and iii) the tumour specific antigen MAGE-A3.

Cells are separated into lymphocytes (and monocytes) by elutriation in a closed system (Elutra). [The fraction richest in lymphocytes contained > 90% lymphocytes and viability was > 95%.]

Cells from healthy blood donors are separated into monocytes (and lymphocytes) by elutriation in a closed system (Elutra). [The two most monocyte-rich fractions collected contained > 80% monocytes, viability was > 95%.]

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

The mice were generated with the intention to elucidate in vivo DC developmental regions in steady state and inflammation in situ.

It was found that lymph nodes that drain sites of mature DC or adjuvant inoculation recruited L-selectin- CCR7- effector and memory CD8+ T cells. This cell recruitment required CXCR3 expression on T cells and occurred through high endothelial cells (HEV) in concert with HEV luminal expression of the CXCR3 ligand CXCL9.

MAGE-A3 specific CD4+ T cells were found also in a high percentage of melanoma patients but CD4+ T cells showed an unpolarized or Th2 skewed phenotype. We have optimized the MAGE-A3 peptides to load onto HLA-DR*1101 tetramers.

CEA specific CD4+ T cells were found both in normal donors and in patients with high-grade cervical lesions, pancreas adenocarcinoma and advanced melanoma but CD4+ T cells from the patients compared to normal donors showed impaired IFN-gamma production.

HPV-18 E6 specific CD4+ T cells were found in a high percentage of patients with cervical lesions and we showed that the quality of the response (i.e. the level of IFN-? produced) could predict the clinical outcome after surgery.

We set an in vitro system to study ex-vivo responses able to detect fona fide in vivo primed CD4+ T cells. We investigated spontaneous CD4+ T cell responses to these antigens in normal donors and in patients with high-grade cervical lesions, pancreas adenocarcinoma and advanced melanoma. The antigens were: i) the E6 and E7 proteins of human papilloma viruses, ii) the carcinoembryonic antigen (CEA) and iii) the tumour specific antigen MAGE-A3.