Tumor growth is supported by tumor stroma, such as tumor associated macrophages (TAM) and tumor associated dendritic cells (TADC). We have recently reported that TAM display massive nuclear localization of the p50 NF-kB inhibitory homodimer, which correlates with impaired inflammatory functions.The functional significance of this observation was demonstrated in p50 NF-kB deficient mice, which displayed tumor growth inhibition.

This vaccine was used in patients with colorectal cancer or ovarium cancer. It was well tolerated and robust p53-specific T cell responses were induced.

This patient is undergoing i.v. application of indium labelled E/L-S DC (without antigen loading) in a study of DC migration.

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.

The patients were vaccinated with a vaccine composed by autologous DCs pulsed with apoptotic allogenic prostate carcinoma cell line LNCap.

We have conducted a phase I long peptide vaccination trial with p53 peptides in Montanide adjuvant in patients with colorectal cancer or ovarium cancer.

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.

We have conducted a phase I long peptide vaccination trial with p53 peptides in Montanide adjuvant in patients with colorectal cancer or ovarium cancer.

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.

These patients were treated in a long peptide trial. Analysis of the local immune response demonstrated the presence of HPV16-specific Th1/Th2 cells infiltrating the vaccination site and the VIN lesion after vaccination. 5 out of 20 patients showed complete clearance of all VIN III disease and in 4 of these this was also associated with complete clearance of HPV16 virus.

PBMC were isolated from buffy coat. Subsequently, monocytes were isolated and differentiated into dendritic cells.

PBMC are Leukapheresis derived and are loaded with peptide pools, each consisting of 10 15-mers, which overlap with 11 aminoacids.

PE-alpha DC80 is used for staining of incubated cells for FACS analysis for activation markers.

50 U/ml of penicillin were added to the monocyte cell culture.

This vaccine consists of G4-DC loaded with 8 different peptides. It was tested in a trial in small cohorts of patients (3-9) who were vaccinated with differently composed DC-vaccines.

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.

Dendritic cell (DC) derived-exosomes (Dex) were described as nanovesicles harbouring functional MHC molecules stimulating T cell responses. 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 in an anti-metastatic effect. In humans, Dex bear functional IL-15R? which allow proliferation and IFNg secretion by NK cells. In contrast to immature DC, human Dex harbor NKG2D ligands on their surface leading to a direct engagement of NKG2D and NK cell activation ...

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

Perforine expressing T cells were observed in human Hemato-Lymphoid System Rag2-/-gc-/- mice after being infected with EBV and mounting an immune response. They infiltrated in B cell infected areas in lymphoid organs in situ.

Cells (co-cultured and frozen stained GM-DC, D2SC1/Flt3-DC and dead cells) were trypsinzed and fixed in 4 % PFA (or formalin).