A long-standing collaboration with Centre for Ecology & Hydrology (CEH, UK) has led to the identification of an activity in salivary gland extracts of the tick Rhipicephalus appendiculatus that profoundly modulates the responses of human monocyte-DC towards extrinsic stimuli. Recombinant protein has been generated from the gene for this molecule (Japanin) which had been cloned. This material inhibits the up-regulation of costimulatory molecules in response to certain TLR agonists, and some cytokines but not others, and modulates the function of the DC in ...

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.

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 protein vaccines were injected combined with a combination of iNKT cell-agonist alpha-GalCer and MPL, a detoxified version of LPS that signals through TLR4. This combination treatment stimulates antigen-specific T and B cell responses that are greater than those elicited with alpha-GalCer or MPL alone.

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

The poly(I:C) was used for stimulating DCs. It was shown in mice that both IL-10 as well as IL-12 production is dependent on the signalling adaptor molecules either MyD88 or TRIF, respectively in response to CpG, LPS or PolyIC. Manufacturer: Invivogen Life Technologies

This PSA was used as a control in the transfection of immature DC. Transfection of immature DCs was done with different constructs encoding for the oncogene Her-2/neu or an adeno virus Her-2 full length construct.

The CpG was used for stimulating DC. It was shown in mice that both IL-10 as well as IL-12 production is dependent on the signalling adaptor molecules either MyD88 or TRIF, respectively in response to CpG, LPS or PolyIC.

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...

This PGE2 was used in a DC maturation process with the final aim to produce a DC vaccine.

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.

PE-alpha DC80 is used for staining of incubated cells for FACS analysis for activation markers.

50 U/ml of penicillin were added to the monocyte cell culture.

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

This vaccine was used in patients with colorectal cancer or ovarium cancer. It was well tolerated and robust p53-specific T cell responses were induced.

The vaccine for ovarian carcinoma is composed by autologous DCs pulsed with apoptotic autologous ovarian carcinoma cells. Apoptosis is induced by UV.

This vaccine consists of G4-DC loaded with 8 different peptides. It was tested in a trial in small cohorts of patients (3-9) who were vaccinated with differently composed DC-vaccines.

The expression and function of these murine FcgammaR in CD11c+CD11b-B220+ plasmacytoid DCs (pDCs) was investigated. pDCs express mostly FcgammaRIIB while the expression of FcgammaRI and FcgammaRIII is only detected by RT-PCR at low but significant level. Moreover, the ITAM-containing intracellular chain associated to FcgammaRI and FcgammaRIII is strongly expressed in pDCs as detected by biochemical assay.

These multimers were provided by P. Coulie and used to establish routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry.

This is mRNA encoding CD40L, CD70, caTLR4 and one tumorantigen (gp100, Tyrosinase, Mage-C2 or Mage-A3). It was used to electroporate autologous dendritic cells in order to produce a vaccine for advanced melanoma patients.