We have chemically coupled the R-epimer of the Pam3CSK4 to long peptides containing a CTL epitope and used this structure to investigate the behaviour of the diastereomer.

We have prepared two plasmids coding for LACK linked to either eGFP or mCherry fluorescent proteins, under the control of the beta-casein promoter. In order to identify the APCs that were responsible for the development of this state of immune tolerance, we aimed to generate a transgenic mouse that express a recombinant fluorescent protein in the mammary glands, an approach that should result in high levels of expression of the fluorescent protein in the milk. We thus should be able to detect the cells that capture, degrade and present this antigen to T cel...

We have prepared several new constructs for the expression of c-type lectin receptors in DC.

We have prepared several new constructs for the expression of (modified) adhesion receptors in DC.

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

In the process of total RNA extraction from yeast cells, the supernatants were precipitated in RNase-free centrifuge tube by adding 1/10 volume 3 M NaAcetate (pH 5.3) and 2.5 volumes of cold ETOH 100%.

In the process of total RNA extraction from yeast cells, the supernatants were precipitated in RNase-free centrifuge tube by adding 1/10 volume 3 M NaAcetate (pH 5.3) and 2.5 volumes of cold ETOH 100%.

In the process of extracting total RNA from yeast cells, the pellets were washed with 70% Ethanol two times.

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

4 ml of chloroform (Fischer BP1145-1) were added to buffered and incubated S. cerevisia cells in a process of total RNA extractions.

2 ml of acid phenol were added to the buffered and incubated S. cerevisia cells after transferral of the supernatant to a new tube.

In a process of total RNA extraction from S. cerevisia, 2 ml of preheated acid phenol (pH 4.3) were added to yeast cells that had been collected and the pellet resuspend in 3.6 ml AE Buffer (AE=50mM NaAcetate pH 5.2, 10mM EDTA).

The melanin-free total tumor RNA is obtained from melanoma metastases; it is suitable for in vitro amplification and DC transfection.

One volume fresh medium containing 2x concentrated cytokines was added to the cell culture of monocytes in serum free medium in the presence of 100 ng/mL GM-CSF (Leukine, Berlex) and 20 ng/mL IL-4 (Cell Genix).

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).

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).

Src kinases were found to be required for the initiation of some maturation characteristics of human monocytes derived dendritic cells upon stimulation with several toll like receptor (TLR) agonists but not of others, since their inhibition was able to dissociate cytokines and chemokines production from the induction of surface maturation markers.

We are examining the effects of a schistosome-derived protein on intestinal immune responses, and have shown that oral administration leads to the rapid appearance of cytokines in lymph.

We are characterising (in collaboration with Sebastian Amigorena) rat lymph exosomes and will determine if they transport intestinally-delivered scrapie ME7.

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...