This DC vaccine was produced from the Leukapheresis product, after elutriation, of a metastatic lesion of stage III/IV melanoma patients.

This DC vaccine was produced from matured DC (matured using TNF alpha, IFN alpha and PGE2) which were subsequently transfected with CD40L-encoding RNA and tumor RNA.

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.

This vaccine was used for RCC patients who were deficient in T cell IFN-gamma and IL-2 production pre-vaccination.

Constructs for fusion proteins of the DC specific receptor DC-SIGN fused to GFP have been completed and tested.

DMSO was used to dissolve blue formazan particles.

The model antigen OVA was fused to the C1C2 exosome-binding domain of MFG-E8/lactadherin. This construct was used in a study on the capture of antigen-bearing exosomes by DCs and cross-presentation of tumour antigen in exosomes.

Total RNA was extracted from yeast cells after buffering, incubation and centrifugation by phenol extraction.

Fabs that bind strongly to L-SIGN, but to a lesser degree or not at all to dendritic cell-specific ICAM-grabbing nonintegrin (DC-SIGN) were isolated and characterized. We believe that the isolated Abs may be useful for selective delivery of Ags to DC-SIGN- or L-SIGN-bearing APCs for the modulation of immune responses and for blocking viral infections.

The FC-block was used for incubation of cells (1:50 in PBS + 2 % FCS) for FACS staining of activation markers.

Monocytes were transferred into culture bags by sterile welding and cultured for 5 days in serum free medium (CellGro, Cell Genix) in the presence of 100 ng/mL GM-CSF (Leukine, Berlex) and 20 ng/mL IL-4 (Cell Genix).

Melanoma patients were vaccinated with 4 peptides (MAGE-A3, gp100, NA17 and tyrosinase) presented by HLA-A2 and monitored. We measured responses to these peptides injected with Montanide, but did not know whether the Montanide adjuvant was important for these T cell responses. 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.

Differentiation of monocytes into dendritic cells is promoted, along with recombinant IL-4, by addition of granulocyte-macrophage colony-stimulating factor (GMCSF 1000U/ml, Chemicon).

We have worked to strengthen vaccine developments with an improved humanized mouse model. We focus on recombinant vaccines against HIV driven by a powerful poxvirus vaccine system termed MVA BN. To improve our understanding MVA BN has been analyzed thoroughly on the nucleotide level, safety in cell cultures, immune deficient mice as well as in healthy, HIV–infected and other immune deficient humans.

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.

Patients with resected HPV16-positive cervical cancer were vaccinated with an overlapping set of long peptides comprising the sequences of the HPV16 E6 and E7 oncoproteins emulsified in Montanide ISA-51.

Patients with resected HPV16-positive cervical cancer were vaccinated with an overlapping set of long peptides comprising the sequences of the HPV16 E6 and E7 oncoproteins emulsified in Montanide ISA-51.

These dimers were injected into TCR transgenic mice which exhibited an increased frequency of LACK-specific T cells with the aim of producing a mAb reacting to the immunodominant LACK156-173 peptide of leishmania bound to I-Ad MHC class II molecules.

We have investigated the respective role of IL-10 and IDO, both required for Th1 suppression in this model. As IL-10 has been shown to indirectly dampen Th1 responses through the inhibition of IL-12 production by antigen-presenting-cells, we monitored mRNA expression specific for the inducible chain IL-12 p35 in lymph nodes draining the site of injection or in mesenteric lymph nodes of mice instilled intra-colonically with TNBS. Our results clearly show that IL-12 production early after the onset of the response was unaffected by anti-CTLA-4 treatment. By cont...

IFN alpha was used in a DC maturation process with the final aim to produce a DC vaccine.