The D2SC1/Flt3-DC were stained (Mini67-1KT or Mini26-1KT (PKH67/PKH26 green/red fluorescent cell linker mini kit for general cell membrane labelling) from Sigma) for a process of dead cell uptake.

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.

These Treg were induced after treatment of primed mice with intact anti-CTLA-4 antibodies. These regulatory T cells inhibit Th1 responses, in vitro and in vivo, and repress experimental intestinal inflammation, by a mechanism involving IL-10 and IDO.

We have investigated the in vivo capacity of resting (and of DC-primed) NK cells to reach the draining lymph nodes.

The capacity of splenic DC, pulsed in vitro with protein antigens or peptides, to induce immunity was monitored after transfer into syngeneic animals. To improve existing strategies, the amplitude and polarisation of T cell responses was monitored in the presence or absence of regulatory T cells.

The kinetic of the transcriptional profiling of purified mouse DC treated with IL10 was analysed.

These DC were generated from monocytes after elutriation of leukapheresis product (patients with stage III or stage IV melanoma) and pulsed with irradiated tumor cells (apoptotic bodies) prepared from the removed lesion and matured for 48 hours in presence of TNF-alpha, and subsequently frozen.

A portion of the DCs is pulsed with irradiated tumor cells (apoptotic bodies) prepared from the removed lesion and matured for 48 hours in presence of TNF-alpha.

RCC product that was tested in an RCC phase 2 trial. The product does indeed lead to restoration of both IL-2 and IFN responses and leads to a gradual decline of the immune effector T cells in the peripheral circulation (i.e., weaker signal after the fifth dose). Using the PME-CD40L DC product we have observed tumor regressions in a number of patients in the current study. At present, 6 of 12 patients that have been restaged using RECIST criteria have less tumor burden that when they enrolled in the study (15 weeks earlier).

For a transcriptional profiling of dendritic cells, DC were stimulated with PolyI:C, pretreated or not with PP2, and microarray analysis was performed.

We have studied the immuno-physiology of rat and mouse intestinal dendritic cells. We have used a known adjuvant, E. coli heat-labile toxin (Etx) and a potential adjuvant R-848, a small TLR7/8 ligand.

These primed T cells have been obtained from untouched naive CD4+ T cells co-cultured with fungi -pulsed DCs.

Purified CD8 T cells using MACS cell separation were co-cultured with electroporated DC and subequently became evaluated on their cytokine secretion, cytotoxicity and % of antigen specific T cells.

We examined whether plasmacytoid pDC are infected and/or activated by MTb, and whether they play a role in regulating bacterial clearance during acute MTb infection and if so how. We have compared these DC with myeloid DC and macrophages with respect to MTb infection.

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.

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.

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.

We have demonstrated that the NADPH oxidase NOX2 is recruited to DC early phagosomes mediating a sustained production of low levels of reactive oxygen species and causing a maintained alkalynization of the phagosomal lumen. DCs lacking NOX2 show increased antigen degradation due to an enhanced phagosome acidification. As a result, the efficiency of in vitro and in vivo antigen cross presentation is significantly reduced in NOX2-deficient DCs.

We found that CD8alpha- DC produced undetectable levels of IL-12p70 upon stimulation with CpG.