Alternatively activated DC (AA-DC) were obtained by culturing immature DC in the presence of maturative agonists (such as, LPS, CD40L or TNF) and calcitriol, IL-10 or prostaglandin E2 for a study of molecular signatures for alternatively activated DC. AA-DC showed a decrease in the production of IL-12 but retained most of the ability to secrete other cytokines, such as TNF and CXCL8. The hallmark of AA-DC was the production of the angiogenic cytokine VEGF in vitro, and in vivo when the cells were implanted into the chicken embryo CAM assay.

These dendritic cells were generated from monocytes isolated from buffy coats, activation was induced by lipopolisaccaride (LPS 1microgram/ml, Sigma, St. Louis, MO), by Curdlan (100 micrograms/ml, Wako), by R848 and yeast RNA and by yeast cells in different conditions of culture.

These adherent peripheral blood monocytes are produced from leukapheresis from HLA-A1 and/or HLA-A2 positive patients with stage III/IV melanoma, used to generate DCs from. The latter will subsequently be used to vaccinate the HLA-A1 and/or HLA-A2 positive patients.

In our effort to demonstrate that pathogen-derived signals to DC mediated via TLRs can be modulated by activated iNKT cells, these DC isolated from animals treated simultaneously with TLR and iNKT cell ligands were potent stimulators of naive T cells in vitro compared with DC from animals treated with the ligands individually.

In our effort to demonstrate that pathogen-derived signals to DC mediated via TLRs can be modulated by activated iNKT cells, DC isolated from animals treated simultaneously with TLR and iNKT cell ligands were potent stimulators of naive T cells in vitro compared with these DC from animals treated with the ligands individually.

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...

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.

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.

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.

After using frozen MoDC for tumor peptide specific stimulation of autologous T lymphocytes at a ratio of 1 DC per 10 T cells, co-cultures led to antigen specific, IFN-? secreting T cells (1 restimulation at a DC:T cell ratio of 1:20).

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. The studies demonstrated for the first time the existence of a bidirectional activating interaction between DCs and gamma delta T lymphocytes activated by aminobiphosphonates. These data suggest a potential adjuvant role of this early cross-talk in the therapeutic activity of aminobiphosphonate drug that are currently used as anti-tumor drugs.

Autologous DC were pulsed with apoptotic allogenic prostate carcinoma cell line LNCap to produce the advanced prostate carcinoma vaccine.

These carcinoma cells were used to pulse autologous DCs for vaccine production for ovarian carcinoma patients.

These cells were used to produce a vaccine for prostate carcinoma and were pulsed with apoptotic allogenic prostate carcinoma cell line LNCap cells.

We evaluated the capacity of autologous DC and HEK293 cells transfected with relevant HLA alleles to function as T cell targets in Elispot assays upon transfection with tumor antigen encoding plasmids. Due to strong background from the autologous DC we decided that HEK293 transfected with one HLA-allele at a time plus simultaneously transfected with up to 5 tumor antiges would be optimal to screen for antigen specificity in the patients T cells. However, during a second T cell culture to expand large numbers of T cells for this screening procedure a drastic ex...

The 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, are used for vaccination.

These autologous DC were electroporated with mRNA encoding CD40L, CD70, caTLR4 and one tumorantigen (gp100, Tyrosinase, Mage-C2 or Mage-A3) to produce a vaccine for immunotherapy of advanced melanoma patients.

These autologous dendritic cells were used for vaccine production. They were pulsed with apoptotic autologous ovarian carcinoma cells.

These autologous DC were used in a phase IB/II study of immunotherapy. Patients were randomized 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).

The T blasts are used to immunize MAGE-3 positive patients.