Zymolyase was used to treat DC, which were used in a study of microrganism survival following uptake by DCs.

Yeast RNA (along with R848, Curdland, LPS and yeast cells in different conditions of culture) was used to induce DC activation.

In a spore culture, zymolyase (2 mg/ml) was used to digest ascum and liberate spores. We initially performed experiments to evaluate zymoliase sensitivity of asci for this specific strain and we assessed that 15 minutes incubation was the minimum time needed to damage the ascus, without touching the spore cell wall. Zymoliase was inactivated by heat (65°C for 2 minutes) and washed away carefully together with the remainings of the empty ascus, by resuspending twice the spore culture in 500 µl of distilled water centrifuging and discarding the supernatant; th...

Vectors were engineered to express distinct chemokines at the local skin site of mice. In particular, attention has been focussed on the recruitment of distinct subsets of DC to skin.

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.

Used for stimulation in the evaluation of the role of Src kinases in DCs during synergic engagement of TLR8 with either TLR4 or TLR3.

TLR ligands (simultaneous activation of TLR4 and TLR8 signaling cascades) were used as stimuli for MDDC and resulted in a marked inhibition of the secretion of the proinflammatory chemokine CCL2 with respect to stimulation through a single TLR.

We have shown recently that the T cell stimulatory capacity of DCs pulsed with tumorantigen-derived peptides can be considerably increased by activating the DCs through electroporation with CD40L, CD70 and constitutively active TLR4 encoding mRNA (TriMix DCs). This vaccine will also be tested in the multicentre trial.

Used for stimulation in the evaluation of the role of Src kinases in DCs during synergic engagement of TLR8 with either TLR4 or TLR3.

Commercially available siRNA Smart Pool from Dharmacon targeted against the STAT3 transcription factor have been successfully transfected in human MDDCs.

Used for stimulation in the evaluation of the role of Src kinases in DCs during synergic engagement of TLR8 with either TLR4 or TLR3.

These conjugates were tested in vivo by either direct vaccination or ex vivo DC loading prior to vaccination. Both approaches show similar effectiveness in CTL priming.

This tumor RNA was used to transfect matured DC with the final aim to produce a DC vaccine.

TNF alpha was used for DC maturation with the final aim to produce a DC vaccine.

TNF-alpha is used during maturation of the DCs.

The peptides are used in the generation of monocyte-derived DC (pulsing of the cells at a concentration of 5 µg/mL in approximately 5 mL and at a cell density of 20 x 10e6/mL for 1 h.).

These TcR-Tg clones are specific for the P14-GP33 antigen. In the individual T cells, we studied the expression of twenty different genes either mediating effector functions, or coding for different receptors involved in T cell differentiation and memory generation.

The TcR-Tg clones are specific for the 0T-1-OVA antigen. In the individual T cells, we studied the expression of twenty different genes either mediating effector functions, or coding for different receptors involved in T cell differentiation and memory generation.

It was evaluated whether STAT-1 was phosphorylated in DC following activation with TLR-dependent Th1 and Th2 stimuli and we found that only in presence of TLR stimuli able to induce Th1 responses STAT-1 was activated.

10 mg/ml sulfo-NHS-LC-biotin (Sigma-Aldrich) in 50 mM NaHCO3 pH 8.5 were used to biotinylate yeasts / spores for 2 hours at 4°C. The remaining reactive biotin molecules were inactivated by incubation in 100 mM Tris-HCl pH 8.0 for 40 minutes at 4°C. DCs were then treated with biotinylated spores/yeasts.